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    13 May 2012, Volume 16 Issue 20 Previous Issue    Next Issue
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    Comparison of osteogenesis between the recombinant human vascular endothelial growth factor 165/recombinant human bone morphogenetic protein 2/deproteinized bone and deep frozen bone 
    He Sheng-jiang1, Zheng Hua1, Ni Wei-dong2
    2012, 16 (20):  3611-3615.  doi: 10.3969/j.issn.1673-8225.2012.20.001
    Abstract ( 240 )   PDF (357KB) ( 332 )   Save

    BACKGROUND: Artificial bone made by vector, osteoinductive factor and growth factor has been proved as an ideal biomaterial.
    OBJECTIVE: To compare the revascularization and osteogenesis ability of the composite of recombinant human vascular endothelial growth factor 165/recombinant human bone morphogenetic protein 2/deproteinized bone (rhVEGF-165/rhBMP-2/DPB) with deep frozen bones. 
    METHODS: All of the left forearm radial bones were manufactured into the models of bone defect about 15 mm. The models were randomly divided into two groups. The rhVEGF-165/rhBMP-2/DPB composite was implanted in the experimental group and the deep frozen bones were implanted in the control group.
    RESULTS AND CONCLUSION: At 16 weeks, the bone in the defect area healed and the density of the grafts was close to the surrounding normal bone tissue in the experimental group; In the control group, more callus formation could be seen between the ends of the defects, and the density of the grafts was higher than that of the surrounding normal bone tissue. In the experimental group, there were no boundaries on the interface of graft-receptor, and the bone healed. In the control group, the boundaries were vague, and part of the bone healed. At 3 days and 1, 2, 4 and 8 weeks, ink perfusion microvascular analysis showed that the vascular formation in the experimental group was more than that in the control group (P < 0.01); Biomechanical testing showed that the three-point bending stress load of the experimental group was significantly stronger than of the control group    (P < 0.01). The composite of rhVEGF-165/rhBMP-2/DPB can induce callus formation, accelerate the vascularization of the graft, and have fair biological and biomechanical function.

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    Osteogenic and angiogenic effects of endothelial cells and osteoblasts co-cultured at different ratios 
    Hao Zeng-tao1, Feng Wei2, Hao Ting2, Yu Bin1
    2012, 16 (20):  3616-3619.  doi: 10.3969/j.issn.1673-8225.2012.20.002
    Abstract ( 251 )   PDF (369KB) ( 303 )   Save

    BACKGROUND: Recent studies have shown that co-cultured endothelial cells and osteoblasts are mutually promotive in bone tissues repair.
    OBJECTIVE: To observe the effects of co-culture of endothelial cells and osteoblasts at different ratios on vasculogenesis and bone formation.
    METHODS: Human umbilical vein endothelial cells and osteoblasts MG63 were isolated and cultured. Endothelial cells were co-cultured with osteoblasts at ratios of 1:0, 8:1, 4:1, 1:1, 1:4, 1:8, 0:1 in a 24-well plate, and monolayers of endothelial cells and osteoblasts were cultured alone respectively as controls. At 7, 14, 21 days after co-culture, cells growth condition, vascular endothelia growth factor level and alkaline phosphatase activity were observed and detected in each group.
    RESULTS AND CONCLUSION: Cells grew well in each group in 21 days. At day 14, in the 1:8 group, osteoblasts began to cluster together. At day 21, the 1:4 group showed significantly more calcification nodules formed by Alizarin red staining than the 1:8 group, and no calcification nodules were seen in osteoblasts cultured alone. Alkaline phosphatase activity in the 1:4 group was significantly increased at the three time points (P < 0.05). The level of vascular endothelia growth factor in the 1:4 group began to increase at day 7 and reached its peak at day 21. These findings suggest that angiogenic effect is more effective when endothelial cells are co-cultured with osteoblasts at the ratio of 4:1, and osteogenic effect is more effective at the ratio of 1:4.

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    Mineralization reaction during osteogenic differentiation of myoblasts stimulated by bone morphogenetic protein 2***☆
    Zhang Li1, 2, Wang Wei2
    2012, 16 (20):  3620-3625.  doi: 10.3969/j.issn.1673-8225.2012.20.003
    Abstract ( 278 )   PDF (352KB) ( 319 )   Save

    BACKGROUND: In recent years, it has been confirmed by a variety of ways that myoblasts can differentiate into osteoblasts under the induction of recombinant human bone morphogenetic protein 2 (rhBMP-2).
    OBJECTIVE: To explore the mineralization reaction during the osteogenic differentiation of myoblasts under the induction of recombinant rhBMP-2 and the feasibility of osteogenic phenotype expression by in vitro induction.
    METHODS: Myoblasts were isolated and harvested from neonatal Wistar rats using differential velocity adherent technique and trypsinization method. After in vitro culture, purification and identification, myoblasts at passage 3 were induced by a medium containing rhBMP-2 for 21 days. Myoblasts in the control group were cultured in vitro in complete medium without rhBMP-2 for 21 days.
    RESULTS AND CONCLUSION: After rhBMP-2 induction, myoblast proliferation gradually slowed down. A small quantity of opaque secretory granules were found in the cytoplasm on day 8 after induction; the number of opaque secretory granules increased on day 14 after induction; and a great quantity of opaque secretory granules were found in the cytoplasm on day 21 after induction while the myoblasts without induction fused into contractile myotubes. The alkaline phosphatase activity of the induced myoblasts increased as time extended; myoblasts reacted positively in the alkaline phosphatase staining, immunochemical staining for type Ⅰ collagen and calcium node staining on day 21 after induction. These findings suggest that mineralization reaction is found in rat myoblasts by rhBMP-2 induction and myoblasts can differentiate into osteoblasts under certain inducing conditions in vitro.

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    Effect of mechanical centrifugal force on the signaling pathways of osteoblast bone morphogenetic protein
    Chen Ming1, Guan Jian1, Duan Feng1, Zhu Li-ping2, Yue Chang-jun1, Wang Jian-bo1
    2012, 16 (20):  3626-3629.  doi: 10.3969/j.issn.1673-8225.2012.20.004
    Abstract ( 301 )   PDF (1096KB) ( 355 )   Save

    BACKGROUND: Runx-2 is the target for the bone morphogenetic protein signaling; signal transduction pathway of bone morphogenetic protein involves in the physiological response process of osteoblasts to the stimulation of mechanical centrifugal force. And it plays an important role in the osteoblast information transfer cascade reactions caused by mechanical signals.
    OBJECTIVE: To investigate the effect of mechanical centrifugal force on the signaling pathway of osteoblast bone morphogenetic protein.
    METHODS: MC3T3-E1 cells were pretreated with Dulbecco's modified Eagle’s medium containing 10% fetal bovine serum for 24 hours, and then divided into control group, 90 r/min group, 180 r/min group and 250 r/min group according to the rotational speed. Each group was divided into three subgroups: 6 hours subgroup, 12 hours subgroup and 24 hours subgroup according to the stimulation times of entrifugal force. The control group was treated in the same environment without centrifugation. The total RNA of each group was extracted, and reversed transcripted to cDNA. The expression of Runx2 was analyzed by quantitative real-time PCR.
    RESULTS AND CONCLUSION: With the extension of applying time, the expression of the Runx-2 mRNA increased; it has a positive correlation with the applying time. The expression of the Runx-2 mRNA in the 180 r/min group was significantly higher than that in the 90 r/min group and 250 r/min group ( P < 0. 05); the expression of the Runx-2 mRNA in the 90 r/min group and   250 r/min group were slightly higher than that in the control group (P=0.119). The results show that different size of centrifugal force and different duration of centrifugation has different effects on the physiological responses of osteoblast bone morphogenetic protein signaling pathway. It plays an important role in the osteoblast information transfer cascade reactions caused by mechanical signals.

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    Ultrasonic testing of tendon-bone repair in rabbits with postreconstructive acute ruptured rotator cuff under stress
    Li Sen1, Jin An-min2, Zhang Hui2, Min Shao-xiong2, Wang Qing1
    2012, 16 (20):  3630-3633.  doi: 10.3969/j.issn.1673-8225.2012.20.005
    Abstract ( 290 )   PDF (356KB) ( 284 )   Save

    BACKGROUND: Pervious studies have showed stress can promote tendon-bone repair, but studies related to ultrasonic testing during tendon-bone repair are few.
    OBJECTIVE: To study the changes in ultrasonic testing of tendon-bone in rabbits with postreconstructive acute ruptured rotator cuff under stress.
    METHODS: Adult male New Zealand white rabbits were selected for establishing animal models of tendon-bone repair with postreconstructive acute ruptured rotator cuff under stress, and then were randomly divided into two groups: stress group and non-stress group. Rabbits in the stress group were trained at 2 weeks after operation. Non-stress group was with caging and free.
    RESULTS AND CONCLUSION: At 2 weeks after operation, the echoes at the site of the rabbit supraspinatus tendon-bone were still discontinuous and the signals of inflammatory exudation appeared more evidently than before in the two groups. At 4 weeks after operation, the linear continuous echoes appeared at the site of the rabbit supraspinatus tendon-bone in two groups, which showed more obvious in the stress group than that in the non-stress group. At 6 weeks after operation, the echoes at the site of the rabbit supraspinatus tendon-bone were still discontinuous in two groups, the areas of discontinuous echoes in stress group tendon significantly reduced. At 8 weeks after operation, the echoes at the site of the rabbit supraspinatus tendon-bone were still discontinuous in two groups, the areas of echoes of the tissues combined tendon and bone is more spacious than before. A certain degree of stress stimulation plays an active role in the repairing of postoperative actue ruptured rabbit rotator cuff tendon-bone and ultrasonic testing can detect the dynamic role of tendon-bone repair with non-invasive and accurate.

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    Effects of treadmill running exercise with different intensity on articular cartilage in rats  
    Zhan Li-qiong1, Dang Na2, Gao Mei-qin3, Ni Guo-xin1, 4
    2012, 16 (20):  3634-3638.  doi: 10.3969/j.issn.1673-8225.2012.20.006
    Abstract ( 311 )   PDF (423KB) ( 429 )   Save
    BACKGROUND: Moderate activities are necessary to maintain the normal structure, histological morphology and physiological function of joints. Excessive exercise and immobility lead to articular cartilage degeneration.
    OBJECTIVE: To explore the effects of treadmill running exercise with different intensity on articular cartilage of rat knee joints.
    METHODS: A total of 18 male adult Wistar rats were randomly assigned into three groups: sedentary group, low intensity exercise group and high intensity exercise group. Serum samples were collected to measure matrix metalloproteinase 3 content using enzyme linked immunosorbent assay. In addition, cartilage characteristics were observed by safranin-O staining. Matrix metalloproteinase 3 and type Ⅱ collagen were stained by immunohistochemical staining, and were evaluated by Mankin’s histological grading system. The mRNA expression of matrix metalloproteinase 3 in cartilage was detected by reverse transcription-PCR.
    RESULTS AND CONCLUSION: Compared with sedentary group and low intensity exercise group, significant differences were found in high intensity exercise group in terms of Mankin’s score, matrix metalloproteinase 3 concentrations in the serum and cartilage (P < 0.05). The contents of matrix glycosaminoglycan and type Ⅱ collagen in high intensity exercise group were significantly lower than those in the sedentary group and low intensity exercise group (P < 0.05). There was no significant difference between sedentary group and low intensity exercise group. These results indicate that high intensity exercise can lead to articular cartilage degeneration in rat knees, and exercise-induced cartilage injury may be related to the increasing expression of matrix metalloproteinase 3.
     
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    Effect of stromal cell derived factor-1 on collagen type Ⅱ and matrix metalloproteinase secretion in osteoarthritis
    Wang Guo-liang, Li Yan-lin, Gao Gang, Ma Ke, Chen Wen-dong, Xu Peng, Yang Guang
    2012, 16 (20):  3639-3643.  doi: 10.3969/j.issn.1673-8225.2012.20.007
    Abstract ( 267 )   PDF (386KB) ( 315 )   Save

    BACKGROUND: The stromal cell derived factor-1 (SDF-1)/chemokine receptor 4 (CXCR4) signal pathway plays a key role in the pathogenesis of osteoarthritis.
    OBJECTIVE: To study the effect of SDF-1 on collagen type Ⅱ and matrix metalloproteinase (MMPs) secretion in osteoarthritis.
    METHODS: Forty knee cartilage blocks from osteoarthritis patients who had total knee replacement were selected as experimental group. Forty knee cartilage blocks from patients who had traumatic amputation were as the control group. The samples were put in Dulbecco's modified Eagle’s medium and cultivated for 48 and 96 hours with 79 and 392 μg/L exogenous SDF-1.
    RESULTS AND CONCLUSION: After cultured for 48 and 96 hours, the expressions of collagen type Ⅱ were both positive in the two groups by immunohistochemistry staining; the collagen type Ⅱ degenerated more when it was exposed in high-concentration SDF-1 culture medium with the longer time. There was no significant difference of SDF-1 in SDF-1 culture medium at the same time and concentration between the experimental and control groups (P﹥0.05). The MMPs of the experimental group was released more than that of the control group in high-concentration SDF-1 culture medium (P < 0.05). These showed that SDF-1 could regulate the expression of MMPs and lead to the degradation of collagen type Ⅱ through SDF-1/CXCR4 signal pathways in osteoarthritis. 

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    Proteomic detection of biological markers for rabbit articular cartilage damage
    Li Jian-xin, Yang Liang, Fu Jing-nan, Zhu Lei, Lü Dan, Wang Wen-liang
    2012, 16 (20):  3644-3648.  doi: 10.3969/j.issn.1673-8225.2012.20.008
    Abstract ( 375 )   PDF (454KB) ( 415 )   Save

    BACKGROUND: The specific markers in joints, blood, urine and other body fluids that can reflect the degree of articular cartilage damage can be detected by proteomics. 
    OBJECTIVE: To further verify the application of biological chip technology to find rabbit articular cartilage damage biological markers.
    METHODS: The improved Hulth method was used to establish the rabbit articular cartilage damage model. Postoperative free activities and the injury limb were not fixed, 30 min/d points off twice for 12 weeks. The rabbit knee without any treatment was as normal control. At 4, 8 and 12 weeks after modeling, part of the specimens (such as serum and joint liquid) were collected, and the articular cartilage damage degree was observed and verified; the joint fluid and serum sample of the animals at each time points were put into the PBS Ⅱ-C protein microarray reading machine and detected with CM10 chip.
    RESULTS AND CONCLUSION: The articular cartilage damage model established by improved Hulth method could quite comprehensively reflect the change of articular cartilage damage from the early, middle, and late and decompensate periods. Compared with normal control group, the expression of 3 475 protein in serum specimen was decreased and the expression of    7 558, 15 475 and 33 665 protein was increased in model group; the expression of 7 558 and 33 278 protein in joint fluid specimen was decreased and the expression of 3950 and 16 055 protein was increased in model group. The significant differences protein peak spectrum with the nuclear mass ratio of 3 475, 7 558, 16 884 was expressed in both serum specimen and joint fluid specimen. Part of the differential protein appeared in the specimen may be the articular cartilage damage biological markers.

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    Effect of tensile stress on type Ⅱ collagen and aggrecan expression in rat condylar chondrocytes*★
    Zheng Ru-song1, Yang Zhu-li2, Du Yan-xiao3, Yin Chong-ying1, Jia Ping-ping1, Yuan Xiao2
    2012, 16 (20):  3649-3653.  doi: 10.3969/j.issn.1673-8225.2012.20.009
    Abstract ( 337 )   PDF (377KB) ( 484 )   Save

    BACKGROUND: Changes in extracellular of chondrocyte can reflect influence of external force on temporomandibular joint and adaptability of body to external force.
    OBJECTIVE: To study the effect of cyclic tensile stress on main extracellular matrix of condylar chondrocyte.
    METHODS: The cyclic tensile stress was exposed to the third passage condylar chondrocyte for 0, 1, 6, 12 and 24 hours, respectively, using a Flexcell Strain Unit-5000T system (10% surface elongation, 6 cycles/min). After mechanical loading, total RNA was extracted from the cells harvested from Six-well BioFlex flexible cell Petri Dish, reverse transcribed, and reverse trabscription-PCR was performed to quantify mRNA levels for type Ⅱ collagen and aggrecan.
    RESULTS AND CONCLUSION: Compared with the control group (0 hour group), both type Ⅱ collagen and aggrecan mRNA expression was significantly increased after loading for 6 hours (P < 0.05), but began to decrease since 12 hours, and significantly decreased at 24 hours (P < 0.05). Results showed that cyclical tensile stress stimuli can affect the synthesis of main extracellular matrix of condylar cartilage, i.e. the synthesis was gradually enhanced with prolonged stimulation duration, but significantly inhibited in response to further stress stimuli.

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    Effects of hypoxia inducible factor 1 on angiogenesis in skeletal muscle tissues of rats receiving hypoxia training
    Zou Zhi-bing1, Mao Shu-zhang2, Zheng Lan3
    2012, 16 (20):  3654-3658.  doi: 10.3969/j.issn.1673-8225.2012.20.010
    Abstract ( 266 )   PDF (421KB) ( 369 )   Save

    BACKGROUND: Hypoxia inducible factor 1 is a kind of transcription factor which plays a crucial role in the organism’s adaptation to changing oxygen tensions. It is closely related to the organism resistance to anoxia.
    OBJECTIVE: To detect the expression of hypoxia inducible factor 1 and vascular endothelial CD34 protein expression in skeletal muscle tissues of rats receiving hypoxic training; to explore the effects of hypoxia inducible factor 1 on angiogenesis in skeletal muscle tissues.
    METHODS: A total of 60 healthy male SD rats were randomly divided into six groups: normoxic control group, hypoxic sedentary group, normoxic training group, hypoxic residence plus hyperoxic training group, hyperoxic residence plus hypoxic training group and hyperoxic residence plus hyperoxic/hypoxic training group. Rats in training group were trained with increasing load treadmill exercise 6 days a week for 10 weeks. The exercise volume changed from 15 m/min for 25 minutes in the 1st week to 28 m/min for 50 minutes in the 10th week. Rats in hypoxic groups were trained every Tuesday, Thursday and Saturday under the hypoxia environment equivalent to 1 500 meters above sea level; and lived in low oxygen environments, whose hypoxic degree increased from that equivalent to 1 800 meters in the 1st week to that equivalent to 3 600 meters above sea level in the 10th week.
    RESULTS AND CONCLUSION: Under the low oxygen condition, hypoxia inducible factor 1 had massive protein expression. The expression was more under the low oxygen environment combined with training. While CD34 protein expression was only detected in normoxic training group and hypoxic trainging group. It indicates that hypoxia inducible factor 1 is an important factor in the promotion of angiogenesis in skeletal muscle tissues, but it can produce positive effects only when combined with exercise.

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    Biological activity of vascular endothelial cells of composite tissue cryopreserved with rapid-freezing versus slow-freezing methods  
    Liu Yuan-xin1, Zhang Shu-ming2, Qiao Lin2, Huo Hai-tao2, Zhu Ze-xing2, Yuan Lei2
    2012, 16 (20):  3659-3662.  doi: 10.3969/j.issn.1673-8225.2012.20.011
    Abstract ( 348 )   PDF (318KB) ( 383 )   Save

    BACKGROUND: To recover good blood supply is critical to the cryopreservation and replantation of composite tissues, and different cryopreservation techniques have different effects on vascular activity.
    OBJECTIVE: To compare the activity of vascular endothelial cells of composite tissue following rapid-freezing and slow-freezing methods.
    METHODS: Forty New Zealand white rabbits were randomly divided into control group, cryopreservation for 12 hours group,    3 days group, 7 days group, with 10 in each group. Control group was without any treatment. In the experimental groups, New Zealand white rabbit hind limbs were divided into two subgroups, receiving rapid freezing and slow freezing. The hind limbs in the rapid freezing group were put into liquid nitrogen for 12 hours, 3 days, and 7 days. In the slow-freezing group, the freezing procedure was carried out at 4 ℃, -20 ℃ and -80℃, successively, using programmed frozen. All the groups required cryopreservation for 12 hours, 3 days, and 7 days, and then rapid rewarming. After thawing, pathological changes in the vascular endothelium were examined by optical microscope and immunohistochemical staining.
    RESULTS AND CONCLUSION: The scores on the morphology of vascular endothelial tissues were higher in the control group than the two freezing groups following cryopreservation for 12 hours, 3 days, and 7 days (P < 0.05). The scores on vascular endothelial growth factors were higher in the slow-freezing group than the rapid-freezing group (P < 0.05). These findings indicate that slow-freezing is superior to rapid-freezing to preserve the biological activity of vascular endothelial cells of composite tissue.

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    Expression of atuocrine motility factor in keloid
    Zhang Jun-bo, Ya Zu-meng
    2012, 16 (20):  3663-3666.  doi: 10.3969/j.issn.1673-8225.2012.20.012
    Abstract ( 277 )   PDF (401KB) ( 407 )   Save

    BACKGROUND: Autocrine motility factor (AMF) exhibits an improved expression in tumor cells, and it can enhance cell growth, invasion and metastasis. But keloid has biological characteristics of malignant tumor.
    OBJECTIVE: To know about the expression of AMF in keloid, hypertrophic scar and normal scar tissues.
    METHODS: Each 20 cases of keloid, hypertrophic scar and normal scar were studied, and three kinds of scar tissues were selected. The distribution and relative expression of AMF in the three kinds of scar tissues were assessed by immunofluorescence. Protein and mRNA level of AMF in scar tissues were analyzed by western blot and real-time PCR methods.
    RESULTS AND CONCLUSION: In tissue, protein and mRNA levels, normal scar and hypertrophic scar tissues showed low expression of AMF. In distribution and relative expression, there was no significant difference between the normal scar and hypertrophic scar tissues. However, the keloid tissues showed significantly higher expression of AMF than normal scar and hypertrophic scar tissues in the three levels (P < 0.01). These findings suggest that AMF is highly related to tumor biological characteristics of keloid.

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    Effect of botulinum toxin type A on the proliferation and collagen synthesis of human hypertrophic scar fibroblasts 
    Li Wei-hua1, Li De-shui1, Gao Yu-wei2, Xing Yu-xi1
    2012, 16 (20):  3667-3670.  doi: 10.3969/j.issn.1673-8225.2012.20.013
    Abstract ( 327 )   PDF (296KB) ( 520 )   Save

    BACKGROUND: Local injection of botulinum toxin type A (BTA) around the wound can reduce the scar formation. The scar hyperplasia and contracture can also be inhibited; thereby the scar can become atrophic and flat.
    OBJECTIVE: To investigate the effect of BTA on the proliferation and collagen synthesis at human hypertrophic scar fibroblasts.
    METHODS: Human hypertrophic scar fibroblasts were isolated and cultured in vitro. The cells at logarithmic growth phase were cultured. Dulbecco's modified Eagle medium (DEME) containing BTA (diluted concentration of 0.1 U/L) was used to intervene the cell growth. Control group used DEME containing fetal bovine serum.
    RESULTS AND CONCLUSION: At 1 to 15 days after cells seeding, the hypertrophic scar fibroblasts showed the spindle-like shape with uniform composition and strong proliferation, the fibroblasts proliferated into monolayers, and the hypertrophic cells showed a high degree of consistency in the control group. The cells in the BTA group grew slowly and the arrangement of cells was scattered. The amount of cells in the BTA group was 79.3% of that in the control group. The collagen synthetic rate of BTA group was 48.4% of that in the control group. It is indicated that BTA can inhibit the proliferation and collagen synthesis of human hypertrophic scar fibroblasts.

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    Formation of glial scar in the rat spinal cord after injury
    Huang Kai, Sheng Wei-bin
    2012, 16 (20):  3671-3674.  doi: 10.3969/j.issn.1673-8225.2012.20.014
    Abstract ( 479 )   PDF (335KB) ( 363 )   Save

    BACKGROUND: After spinal cord injury, the treatment cannot completely solve the problem with the body because glial scar forms and cystic degeneration occurs in the spinal cord tissue. Therefore, it is of great significance to know the regularity of glial scar development.
    OBJECTIVE: To analyze the spatial distribution, characteristics of axon and time characteristics of glial scar in the rat spinal cord after experimental spinal cord injury.
    METHODS: SD rats were divided into control group, 1-day group, 3-day group, 5-day group, 1-week group, 2-week group, 4-week group, 6-week group, 8-week group, 10-week group and 12-week group. Allen’s weight-drop method was performed to prepare spinal cord injury models in rats expect the control group.
    RESULTS AND CONCLUSION: At 4 weeks after spinal cord injury, glial scar and smooth cavity wall began to form. No astrocytes positive for glial fibrillary acidic protein and axons positive for neurofilament protein existed inside the cavity. The glial scar begun to stabilize at 4 weeks after spinal cord injury, and mechanical barriers between the cavity and axon came into being. Thickness of the glial scar no more increased at 10 weeks after spinal cord injury.

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    Effects of dragon’s blood on the expression of substance P and Bcl-2 in wound tissue of diabetic scalded rats  
    Zhang Xian-fa1, Wu Zheng-qiu2, Liang Zi-qian1, Zhao Xue-kai1, Ding Hua-rong1
    2012, 16 (20):  3675-3679.  doi: 10.3969/j.issn.1673-8225.2012.20.015
    Abstract ( 401 )   PDF (522KB) ( 441 )   Save

    BACKGROUND: Previous studies have demonstrated that dragon’s blood can promote the wound healing as well as have characteristics of anti-platelet aggregation, anti-inflammatory analgesic, anti-bacterial and antioxidant and other biological activity.
    OBJECTIVE: To identify the effects of dragon’s blood on the expression of substance P and Bcl-2 in wound tissue of diabetic scalded rats.
    METHODS: A total of 112 Wistar rats were randomly divided into four groups: dragon’s blood group, silver sulfadiazine (SD-Ag) group, diabetic control group and normal control group. Diabetic deep Ⅱ degree burn models were established in dragon’s blood group, SD-Ag group and diabetic control group, and the normal deep Ⅱ degree burn models were established in normal control group.
    RESULTS AND CONCLUSION: On the 7th day post scald, the wound healing rates of dragon’s blood group and normal control group were higher than those of SD-Ag group and diabetic control group (P < 0.05). After 21 days, the wound healing rate of dragon’s blood group was significantly higher than that of SD-Ag group and diabetic control group (P < 0.05) and highest in normal control group. After 15 days, the positive expression level of substance P reached a peak in normal control group and was higher than other three groups, and then decreased gradually. The expression was most positive in dragon’s blood group, SD-Ag group and diabetic control group at the 21th day post scald, and which was significantly higher in dragon’s blood group (P < 0.05). On the 3rd day after scald, the expression level of Bcl-2 in the normal control group was significantly higher than that in other groups. From the 15th day, the expression of Bcl-2 was gradually increased in each group and reached to peak, and highest in dragon’s blood group (P < 0.05), but there was no significant difference between dragon’s blood group and normal control group (P > 0.05). The results demonstrate that dragon's blood can promote the healing of diabetic burn wounds significantly by regulating the expression of substance P and Bcl-2.

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    Surface electromyography analysis of the lower limb muscles of normal young people during natural gait   
    Huang Ping, Qi Jin, Deng Lian-fu, Chen Bo
    2012, 16 (20):  3680-3684.  doi: 10.3969/j.issn.1673-8225.2012.20.016
    Abstract ( 581 )   PDF (466KB) ( 1009 )   Save

    BACKGROUND: Bioelectricity activities of the muscle are the function part of the human body, which can reflect the motor function of the human body.
    OBJECTIVE: To observe and analyze the surface electromyography activity of the lower limb muscles of normal young people during natural gait.
    METHODS: Lower limb muscles (Rectus femoris muscle, Tibialis anterior muscle, Biceps femoris muscle, Medial head of gastrocnemius muscle) of 30 normal young people were tested in flat land walking by surface electromyography (NORAXON TELEMYO 2400R G2, USA). The variation regularity of each parameter within every normal gait was analyzed.
    RESULTS AND CONCLUSION: While the normal young people ware walking naturally on flat ground, their muscle electrical activities changed in periodic of active and static along with the gait cycle, and the right left side muscle of the same name moved alternately; The maximal values of the mean amplitude, the integral electromyography, the mean frequency, the median frequency were the medial head of gastrocnemius muscle, others from large to small were Tibialis anterior muscle, Biceps femoris muscle, Rectus femoris muscle, and the right-left leg distributed rule was consistent; The mean amplitude, the mean frequency, the median frequency of the right medial head of gastrocnemius muscle were lower than the left corresponding values, and the difference had statistical significance (P < 0.05); Trained muscle's time domain and frequency range fluctuated in a certain scope. The electrical activity of the lower limb muscles of normal young people is rhythmic and an alternative movement of the right and left during natural gait; The electrical activity of the medial head of gastrocnemius muscle is strongest in trained muscles; The electrical activity of the medial head of gastrocnemius muscle is different between the dominant side and the non-dominant one; Lower limb muscle’s time domain and frequency range can fluctuate in a certain scope.

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    Establishment of a rotator cuff tear model in rabbits and its preliminary histological study
    Li Feng-long1, Jiang Chun-yan1, Lu Yi1, Li Guang-ping3, Lu Yao-jia2
    2012, 16 (20):  3685-3689.  doi: 10.3969/j.issn.1673-8225.2012.20.017
    Abstract ( 389 )   PDF (452KB) ( 542 )   Save

    BACKGROUND: The modal and structural characteristics of supraspinatus in rabbits are similar to human. In the rotator cuff of rabbits there are tendon-bone transitional zones similar to human, and it is appropriate to investigate the tendon-bone healing aspects with a rabbit model. 
    OBJECTIVE: To establish an animal model of rotator cuff tear in rabbits and to investigate the feasibility of this model followed by the initial histological study. 
    METHODS: Supraspinatus tenotomy was performed in the right shoulder of 18 skeletally matured male New Zealand white rabbits to establish the animal model of rotator cuff tear. The rabbits were randomly divided into two groups: repair group, repaired at 1 week after tenotomy; control group, without repair. The rabbits were sacrificed at 2, 4 and 8 weeks postoperatively in each group with three rabbits sacrificed at each time point. Fatty infiltration of supraspinatus and healing of tendon-bone interface were observed by hematoxylin-eosin staining. 
    RESULTS AND CONCLUSION: The gross anatomy and histological characteristics of the supraspinatus in rabbits were similar to human. The animal model provided a stable surgical approach which ensured a highly replicable procedure in the establishment of rotator cuff tear/repair models. There was no obvious fatty infiltration or muscle atrophy in the supraspinatus in both groups. At 8 weeks after repair, we observed the formation of a new enthesis of supraspinatus in the repair group. There was no new enthesis of supraspinatus in the control group. The rabbit is suitable to establish an animal model of rotator cuff tears. The animal model designed in this study could be used in the research of rotator cuff tears. For the acute supraspinatus tear, formation of a new enthesis can be observed at 8 weeks after repair.

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    Establishment of an experimental atherosclerosis model by feeding high-fat diet plus injecting bovine serum albumin in rabbits
    Li Lin, Dou Jian-lin, Chu Tian-shu, Li Wan-qiu, Zhang Ge, Sun Lin
    2012, 16 (20):  3690-3693.  doi: 10.3969/j.issn.1673-8225.2012.20.018
    Abstract ( 244 )   PDF (327KB) ( 387 )   Save

    BACKGROUND: It is commonly seen to establish a model of atherosclerotic stenosis by simply feeding high-fat diet.
    OBJECTIVE: To establish an atherosclerosis model by feeding high-fat diet plus injecting bovine serum albumin in rabbits.
    METHODS: Three different methods were used to establish rabbit models of atherosclerosis, including simple high-fat diet, high-fat diet+lipid milk, and high-fat diet+bovine serum albumin. Normal control group was fed with normal food.
    RESULTS AND CONCLUSION: The levels of serum total cholesterol, triglyceride, low density lipoprotein and high density lipoprotein were significantly increased in the former three groups compared with normal control group (P < 0.01). The rabbits treated with simple high-fat diet possibly presented with hyperlipidemia rather than atherosclerotic lesions. The rabbits treated with high fatty feedstuff only showed hyperlipidemia without atherosclerosis lesion. The rabbits treated with high-fat diet and lipid milk showed fibrous plaque. The mature atherosclerotic plaques were presented in the rabbits treated with high-fat diet+bovine serum albumin. The mature atherosclerotic plaque may be created by high-fat diet plus bovine serum albumin that an ideal model of atherosclerosis can be established in rabbits.

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    Alteration of oligodendrocyte precursor cells in aged rat models after transient cerebral ischemia 
    Li Hua-jie, Wu Jian, Zhu Lin-feng, Tang Xiao-chun
    2012, 16 (20):  3694-3697.  doi: 10.3969/j.issn.1673-8225.2012.20.019
    Abstract ( 221 )   PDF (303KB) ( 254 )   Save

    BACKGROUND: At present, it is not yet clear about the alteration of oligodendrocyte precursor cells after transient cerebral ischemic and in the repair process.
    OBJECTIVE: To investigate the alteration of oligodendrocyte precursor cells in aged rats after transient cerebral ischemia.
    METHODS: A rat model of transient cerebral ischemia was established by thread embolism methods. Twenty-four aged male SD rats were randomly divided into four groups: control group, days 1, 7 and 14 after ischemia-reperfusion groups. Rats in the control group were only subjected to an exposed blood vessel and sutured with no embolism. Rats in the ischemia-reperfusion groups were subjected to reperfusion for 1, 7 and 14 days after model establishment.
    RESUTS AND CONCLUSION: ①Compared with the control group, the number of cells positive for chondroitin sulfate proteoglycan, unmature oligodendrocyte marker and 2’,3’-cyclic nucleotide 3’-phosphodiesterase in the infarction core was significantly decreased at each time point after transient cerebral ischemia. ②In peri-infarction area, the number of cells positive for chondroitin sulfate proteoglycan and unmature oligodendrocyte marker was increased gradually with time, which was increased dramatically at days 7 and 14 after transient cerebral ischemia. While, the number of 2’,3’-cyclic nucleotide 3’-phosphodiesterase-positive cells was decreased slightly at days 1 and 7 after transient cerebral ischemia. ③There was no significant difference among the number of the three kinds of antigen positive cells in the contralateral cerebral infarction area. These findings suggest that the number of oligodendrocyte precursor cells is increased in the peri-infarcted area after transient cerebral ischemia in aged SD rats. The cells may differentiate into mature cells and participate in the repair of cerebral ischemia injury.

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    Expression of protein-tyrosine phosphatase 1B and insulin receptor substrate 2 in the skeletal muscle of rats with insulin resistance induced by high-fat diet   
    Zhao Hui1, Yu Su-guo1, Wang Ling-ling2, Zhao Yan-min1
    2012, 16 (20):  3698-3702.  doi: 10.3969/j.issn.1673-8225.2012.20.020
    Abstract ( 290 )   PDF (496KB) ( 320 )   Save

    BACKGROUND: Insulin resistance in the peripheral tissue is a major cause of type 2 diabetes mellitus.
    OBJECTIVE: To observe the expression of protein tyrosine phosphatase 1B (PTP-1B) and insulin receptor substrate 2 (IRS-2) in the skeletal muscle of rats with insulin resistance induced by high-fat diet.
    METHODS: Twenty SD rats were randomly divided into normal control group with 10 rats and high-fat diet group with 10 rats. Rats in the two groups were fed with normal diet and high-fat diet for 12 weeks, respectively.
    RESULTS AND CONCLUSION: The insulin sensitive index was significantly decreased in the high-fat diet rats compared with the normal rats (P < 0.01). In obese rats, glucose tolerance and the acute first-phase insulin secretory response to glucose were impaired. The protein level of PTP-1B in the skeletal muscle of obese rats was significantly increased compared with the control group (P < 0.01). Insulin-stimulated IRS-2 phosphorylation in the skeletal muscle was reduced in the obese rats (P < 0.01). These indicate that the increased PTP-1B level and the reduced insulin-stimulated IRS-2 phosphorylations in the skeletal muscle seem to play an important role in the insulin resistance induced by high-fat diet.

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    Effect of modified acidic fibroblast growth factor against cardiomyocyte apoptosis in neonatal rats after anoxia/reoxygenation
    Li Yan1, Wang Yong-ling1, Li Xiao-kun2, Li Jing3
    2012, 16 (20):  3703-3706.  doi: 10.3969/j.issn.1673-8225.2012.20.021
    Abstract ( 232 )   PDF (361KB) ( 383 )   Save

    BACKGROUND: Acidic fibroblast growth factor (aFGF) can protect cardiomyocytes from ischemia/reperfusion damage, but the proliferative ability of the cells induced by modified aFGF (maFGF) is decreased.
    OBJECTIVE: To study the effect of aFGF and maFGF on cardiomytes apoptosis after anoxia/reoxygenation.
    METHODS: Myocardial anoxia/reoxygenation models with primary cultured cardiomyocytes from neonatal SD rats were established and then treated with aFGF and maFGF. The apoptosis rate of cardiomyocytes was detected by flow cytometry assay and the survival rate of cardiomyocytes was measured by typan blue exclusion.
    RESULTS AND CONCLUSION: Pretreatment with aFGF and maFGF decreased the apoptosis rate of cardiomyocytes after anoxia/reoxygenation (P < 0.01). It has been proved that aFGF and maFGF have notable effects on attenuating cardiomyocyte apoptosis after anoxia/reoxygenation.

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    Effects of selective profound hypothermia on the ultrastructure and Vimentin expression of monkey cerebral hippocampi after severe cerebral ischemia 
    Niu Xiao-qun1, Fan Yao-dong2, Pu Jun3, Li Jiang-xiao4, Gao Yong-jun3, Fu Guo-ping3, Fang Shao-long3, Xu Wei3
    2012, 16 (20):  3707-3710.  doi: 10.3969/j.issn.1673-8225.2012.20.022
    Abstract ( 397 )   PDF (332KB) ( 301 )   Save

    BACKGROUND: Previous studies have shown that cerebral selective profound hypothermia combined with antegrade cerebral perfusion can improve the resistance of monkeys to cerebral hypoxia-ischemia.
    OBJECTIVE: To observe the effect of selective cerebral profound hypothermia on the ultrastructure and Vimentin expression of monkey hippocampi after severe cerebral ischemia.
    METHODS: Eight healthy adult rhesus monkeys were randomly divided into two groups: profound hypothermia (n=5), and normothermia (n=3). For the monkeys in the profound hypothermia group, bilateral carotid arteries and jugular veins were occluded for 10 minutes at room temperature; Ringer’s solution at 4 ℃ was then perfused through the right internal carotid artery and flowed out of the right jugular vein, and the brain temperature remained below 18 ℃; 60 minutes later, the cerebral blood flow was restored. For the normothermia group, all procedures were the same except that the perfusion liquid was replaced with Ringer's solution at 37 ℃.
    RESULTS AND CONCLUSION: All animals in the profound hypothermia group were successfully resuscitated. No significant abnormalities of the hippocampus morphology and ultrastructure were observed. In contrast, in the normothermia group, no monkeys were alive after perfusion, with abnormal hippocampus morphology and ultrastructure to different extents. Vimentin expression in the hippocampus was significantly lower in the profound hypothermia group than the normothermia group (P < 0.01). Selective cerebral profound hypothermia following 10-minute occlusion of the bilateral common carotid arteries could down-regulate Vimentin expression in the hippocampus and protect the hippocampus from severe cerebral ischemia.

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    Effect of reactive oxygen species mediated high glucose concentration on the expression of oncofetal fibronectin in human mesangial cells 
    Du Chao1, Xiong Qin-pan2, Zhou Bo1
    2012, 16 (20):  3711-3714.  doi: 10.3969/j.issn.1673-8225.2012.20.023
    Abstract ( 293 )   PDF (293KB) ( 317 )   Save

    BACKGROUND: Researches showed that oncofetal fibronectin is an important indicator of new synthetic extracellular matrix. But studies of relationship between oxidative stress in high glucose environment and oncofetal fibronection are rarely reported.
    OBJECTIVE: To investigate the effect of high glucose on the expression of reactive oxygen species (ROS) and oncofetal fibronectin mRNA in human mesangial cells.
    METHODS: The cultured mesangial cells were divided into following groups: normal control group (5 mmol/L D-glucose), osmotic control group (5 mmol/L D-glucose+20 mmol/L L-glucose) and high glucose group (25 mmol/L D-glucose); α-lipoic acid intervention group containing high glucose+α-lipoic acid 50 group (25 mmol/L D-glucose+50 μmol/L α-lipoic acid), high glucose+α-lipoic acid 100 group (25 mmol/L D-glucose+100 μmol/L α-lipoic acid) and high glucose+α-lipoic acid 200 group    (25 mmol/L D-glucose+200 μmol/L α-lipoic acid). The expression of oncofetal fibronectin mRNA was detected by reverse transcription-PCR. ROS levels were tested by fluorescence microscopy and fluorescence microplate reader.
    RESULTS AND CONCLUSION: High glucose could promote the production of ROS and increase the expression of oncofetal fibronectin mRNA in the mesangial cells. α-lipoic acid could significantly reduce the level of ROS and expression of oncofetal fibronectin mRNA in the mesangial cells under high glucose loaded, which showed a concentration-dependent manner. It is indicated that ROS mediated high glucose can induce the expression of oncofetal fibronectin in the mesangial cells, but this effect can be partially reversed by α-lipoic acid.

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    Essential role of microRNA-146a in proliferation and apoptosis of vascular smooth muscle cells 
    Xiong Wei1, Dong Shao-hong1, Yuan Jian-hui2, Liu Jian-jun2, Xu Xin-yun2, Li Jiang-hua1
    2012, 16 (20):  3715-3719.  doi: 10.3969/j.issn.1673-8225.2012.20.024
    Abstract ( 279 )   PDF (372KB) ( 414 )   Save

    BACKGROUND: MicroRNA-146a (miRNA-146a) can mediate the proliferation of immune cells and tumor cells through negative regulation of nuclear factor κB signaling pathway, but whether miRNA-146a participates in the proliferation or apoptosis of vascular smooth muscle cells (VSMCs) has not been reported.
    OBJECTIVE: To investigate the role of miRNA-146a in VSMCs proliferation and apoptosis and to exploit its mechanisms.
    METHODS: Artificial synthesized miRNA-146a antisense oligonucleotide (ASO, 50 nmol/L), scramble (control, 50nmol/l) and phosphate buffered saline (normal) were transfected into VSMCs by Lipofectamine 2000 individually.
    RESULTS AND CONCLUSION: By the end of 48 hours of transfection, there were significantly lower levels of miRNA-146a mRNA in ASO treated VSMCs compared with that in normal and control VSMCs (P < 0.01). Meanwhile, ASO treated VSMCs manifested a lower proliferative (P < 0.01) and higher apoptotic ability (P < 0.05). The protein level of nuclear factor-κBp65 and proliferation cell nuclear antigen in ASO treated VSMCs were remarkably lower than that in normal and control VSMCs (P < 0.05). miRNA-146a is capable of promoting proliferation and suppressing apoptosis of VMSCs, which is probably related with the increase in nuclear factor-κBp65 expression.

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    Feasibility of rapidly constructing scaffold-free cell sheets for tissue engineering using orthogonal design method
    Zhang Wei1, Zhang Yu-ping2, Xu Yan-li2, Chen Xiao-yan2
    2012, 16 (20):  3720-3724.  doi: 10.3969/j.issn.1673-8225.2012.20.025
    Abstract ( 315 )   PDF (357KB) ( 290 )   Save

    BACKGROUND: Solid organ transplantation has been actively developed with a good therapeutic effect, but the lack of organ donor source is a worldwide problem.
    OBJECTIVE: To develop a centrifugal cell seeded method for rapid and efficient tissue engineering construction.
    METHODS: Different acceleration forces and cell densities and centrifugal times were studied. The orthogonal design method was used to optimize centrifugation parameters for cell seeding. To test stability and reproducibility of the orthogonal design results, experiments were studied using optimization centrifugation parameters. Monolayer cell adherence ratio after centrifugation and harvested cell numbers after cultured for 3 days was counted. Epithelial stratification tissues were constructed in vitro. The cell sheets were constructed on transwell culture insert using centrifugation parameters. Pathological sections were cut and stained with hematoxylin and eosin for 7 consecutive days.
    RESULTS AND CONCLUSION: The (93.75±9.58)% attached cells were achieved using optimal cell seeding density at 9×105 cells/cm2 with a centrifugation at 2 200 r/min for 4 minutes. Monolayer centrifugal seeded cell sheets showed better cell viability and proliferation compared to naturally seeded cell sheets. The one-layer to four-layer cell sheets were rapidly constructed and showed good viability. Different layers of scaffold-free cell sheets were rapidly constructed using optimal centrifugation procedure. It demonstrates that the centrifugation method is a good way for tissue engineering construction.

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    Shudi Biejia and mesangial cells cooperatively up-regulate the gene and protein expression of osteocalcin in osteoblasts
    Lin Ri-yang1, Wu Feng1, 2, He Li-qun1, 2, 3
    2012, 16 (20):  3725-3729.  doi: 10.3969/j.issn.1673-8225.2012.20.026
    Abstract ( 358 )   PDF (374KB) ( 317 )   Save
    BACKGROUND: Serum containing Shudi and Biejia up-regulates the gene expression of osteocalcin in osteoblasts; the mesangial cells supernatant can also up-regulate the gene expression of osteocalcin in osteoblasts.
    OBJECTIVE: To detect if there are additive effects of the mesangial cells supernatant and serum containing Shudi and Biejia on the gene and protein expression of osteocalcin in osteoblasts.
    METHODS: Primary cultured osteoblasts of SD rats were divided into four groups after identification. Control group contained blank rat serum. Shudi Biejia group contained rat serum containing Shudi Biejia. Mesangial cell supernatant group contained blank rat serum and mesangial cell supernatant. Shudi Biejia plus mesangial cell supernatant group contained mesangial cell supernatant and rat serum containing Shudi Biejia. Culture medium was collected on day 6 after cultivation and was well mixed with the culture medium collected on day 3 after cultivation for detection.
    RESULTS AND CONCLUSION: Compared with the control group, the mRNA expression of osteocalcin in Shudi Biejia group, mesangial cell supernatant group and Shudi Biejia plus mesangial cell supernatant group was significantly up-regulated (P < 0.05); the expression rate of osteocalcin mRNA in the Shudi Biejia plus mesangial cell supernatant group was significantly higher than that in the Shudi Biejia group and mesangial cell supernatant group. The absorbance value of osteoblastic osteocalcin concentration in the Shudi Biejia group and mesangial cell supernatant group was significantly higher than that in the control group (P < 0.05). Compared with Shudi Biejia group, the absorbance value of osteoblastic osteocalcin concentration in the Shudi Biejia plus mesangial cell supernatant group was significantly increased (P < 0.05). These findings indicate that the mesangial cell supernatant and serum containing Shudi Biejia both can increase the gene expression of osteocalcin in osteoblast; the combined action of the two can further up-regulate the gene expression of osteocalcin; meanwhile the osteocalcin secretion also increases significantly.
     
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    Effect of aerobic exercise on the expression of microRNA-133a and myocyte enhancer factor 2 in the gastrocnemius
    Zhang Jia-wei1, Ren Wen-jun2
    2012, 16 (20):  3730-3734.  doi: 10.3969/j.issn.1673-8225.2012.20.027
    Abstract ( 372 )   PDF (377KB) ( 296 )   Save
    BACKGROUND: Studies have demonstrated that mircoRNA-133a, myocyte enhancer factor 2, and myogenic differentiation antigen can regulate the development and remodeling of skeletal muscle.
    OBJECTIVE: To investigate the effect of aerobic exercise on the expression of mircoRNA-133a, myocyte enhancer factor 2, and myogenic differentiation antigen in the gastrocnemius.
    METHODS: Forty male SD rats were randomized into control and training group. The treadmill training was performed in the training group, and no training was in the control group. After training for 4 and 6 weeks, myosin, microRNA-133a, myocyte enhancer factor 2, myoblast differentiation antigen mRNA expressions in the gastrocnemius were detected using quantitative real-time PCR, and the changes in cross-section area of type Ⅱ muscle fiber was detected using immunohistochemistry method.
    RESULTS AND CONCLUSION: After the continuous treadmill training for 4 and 6 weeks, the relative weight of gastrocnemius and the contents of myosin heavy chain II a mRNAs increased significantly compared with the control group (P < 0.05 or P < 0.01). Simultaneously, the cross-sectional area of type Ⅱ muscle fibers increased (P﹤0.05). The expressions of mircoRNA-133a and myocyte enhancer factor 2 increased significantly (P < 0.05 and P < 0.01) in training groups after 4 and 6 weeks of aerobic exercise, while the expression of myogenic differentiation antigen maintained the same levels. It is indicated that aerobic exercise elevates the expressional levels of mircoRNA-133a and myocyte enhancer factor 2.
     
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    Protection effect of Gastrodin on chemical damage of skeletal muscle cells in rats  
    Yang Yu-qin, Zhu Dao-li, Chen Pei-lin, Yu Chun-mei, Yang Yan-hua
    2012, 16 (20):  3735-3738.  doi: 10.3969/j.issn.1673-8225.2012.20.028
    Abstract ( 264 )   PDF (428KB) ( 375 )   Save

    BACKGROUND: Modern pharmacological studies have showed that Gastrodin can restore the imbalance between excitability and inhibition of cerebral cortex, produce sedation, sleep, analgesia and other central inhibition.
    OBJECTIVE: To investigate the protective effect of Gastrodin on chemical damage of skeletal muscle cells in L6 rats.
    METHODS: A chemical myoblast damage rat model of L6 was established by using H2O2. Samples of pre-protective group were cultured in Dulbecco's modified Eagle’s medium (DMEM) containing different concentrations of Gastrodin (1.562 500, 0.781 250, 0.390 625 g/L) for a 24-hour pretreatment, followed by the incubation of H2O2 (0.1 mmol/L) for 80 minutes. At the same time, the proliferation group (different concentrations of Gastrodin for a 24-hour pretreatment, but not followed by the incubation of H2O2), model group (only the incubation of H2O2), positive control group (only L6 rat myoblast suspension), negative control group (only DMEM medium) and control group (without Gastrodin pretreatment and H2O2 incubation) were established.
    RESULTS AND CONCLUSION: 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay showed that the survival rate of the proliferation and pre-protective groups was significantly higher than that of the model group. Cell nuclei were labeled by DAPI turned blue, while the fluorescent intensity of DAPI in the model group was obviously much lower than that in the pre-protective group and so was the quantity of cells labeled by DAPI. Cell nuclei were more regularly shaped, more compact cytoplasm and clearer cell outline in the pre-protective group from hematoxylin-eosin staining. However, cells with structural damages were obviously observed in the model group. Bax and Bcl-2 immunofluorescent cytochemical technique demonstrated that the expression of bax in the model group was evidently higher than that in the pre-protective group, while the expression of bcl-2 was the exact opposite. According to the results of flow cytometry, the rate of apoptosis in the pre-protective group was significantly lower than that in the model group (P < 0.05). It is indicated that Gastrodin has remarkable effect on injury of skeletal muscle cells in rats induced by chemicals.

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    Protective effect of Astragalus membranceus polysaccharide on skeletal muscle ischemia-reperfusion injury
    Li Jian-guo, Meng Zhuang-zhi, Liu Hai-ying, Tian Geng, Bi Fu-long, Li Di
    2012, 16 (20):  3739-3742.  doi: 10.3969/j.issn.1673-8225.2012.20.029
    Abstract ( 347 )   PDF (327KB) ( 303 )   Save

    BACKGROUND: Previous studies have demonstrated that Astragalus membranaceus can influence the apoptosis and ischemia-reperfusion injury of myocardial cells. However, it is unclear whether Astragalus membranaceus plays the same role in skeletal muscle ischemia-reperfusion injury.
    OBJECTIVE: To study the protective effect of Astragalus membranceus polysaccharide on skeletal muscle ischemia-reperfusion injury.
    METHODS: Adult rabbits were randomly divided into experimental and control groups, and a model of limb ischemia-reperfusion injury in rabbits was established. Normal saline injection and Astragalus membranceus polysaccharide were infused intravenously into the rabbits of control and experimental groups, respectively, when blood perfusion was about to be recovered. At preischemia, 2 hours post-ischemia, 1, 3 hours post-reperfusion, blood samples were collected from the operated femoral veins to measure the activities of lactic dehydrogenase and creatine kinase. Meanwhile, the tibialis anterior tissues at the above time points were selected to measure the wet/dry weight ratio of skeletal muscle tissues, as well as to observe fine structure and ultrastructure changes.
    RESULTS AND CONCLUSION: After reperfusion, the activities of lactic dehydrogenase and creatine kinase in the experimental group were significantly higher than those in the control group at the same time point (P < 0.01). The wet/dry weight ratio of skeletal muscle tissues in the experimental group was lower than that in the control group at the same time point (P < 0.05). The results of an electron microscope and a light microscope showed that the degree of skeletal muscle injury in the experimental group was slighter than that in the control group. These findings suggest that Astragalus membranceus polysaccharide can relieve tissue swelling and protect skeletal muscle from ischemia-reperfusion injury.

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    Effects of serum containing Lycium barbarum polysaccharides on the intracellular Ca2+ concentration and the expression of collagen type Ⅰ in MC3T3-E1 cells in vitro  
    Ma Feng, Gao Jun, Wang Yi-nong, Shen Jun, Ma Jing-zu
    2012, 16 (20):  3743-3746.  doi: 10.3969/j.issn.1673-8225.2012.20.030
    Abstract ( 310 )   PDF (375KB) ( 426 )   Save

    BACKGROUND: Previous experiments have shown that Lycium barbarum polysaccharide has a remarkable therapeutic effect on osteoporosis in adult castrated female rats.
    OBJECTIVE: To explore the effects of rat serum containing Lycium barbarum polysaccharides on the intracellular Ca2+ concentration and the expression of collagen type Ⅰ in osteoblastic MC3T3-E1 cells in vitro.
    METHODS: The rat serum containing 5%, 10% and 20% Lycium barbarum polysaccharide was added into the media of osteoblastic MC3T3-E1 cells, respectively. Blank serum with 20% volume concentration was added into the control group. 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide viability analysis was utilized to investigate the cell proliferation. The intracellular Ca2+ concentration was determined by laser scanning confocal microscope. The expression of collagen type Ⅰ was examined by immunocytochemical technique.
    RESULTS AND CONCLUSION: Compared with control group, the rat serum containing 5% and 10% Lycium barbarum polysaccharides significantly promoted the proliferation of osteoblastic MC3T3-E1 cells (P < 0.05, P < 0.01). Rat serum containing 10% and 20% Lycium barbarum polysaccharides increased the intracellular Ca2+ concentration and the expression of collagen typeⅠin osteoblastic MC3T3-E1 cells (P < 0.05, P < 0.01). These findings indicate that rat serum containing Lycium barbarum polysaccharides may prevent and heal osteoporosis by affecting the intracellular Ca2+ concentration and the expression of collagen typeⅠ.

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    Effect of different doses of astragalus injection on neuronal apoptosis and caspase-3 expression in the hippocampus of rats after cerebral ischemia reperfusion
    Liu Sha-sha1, Gao Wei-juan2, Qian Tao1, Zhang Xia1
    2012, 16 (20):  3747-3750.  doi: 10.3969/j.issn.1673-8225.2012.20.031
    Abstract ( 346 )   PDF (348KB) ( 354 )   Save

    BACKGROUND: Studies have reported that astragalus injection can inhibit neuronal apoptosis in the hippocampus of rats after cerebral ischemia reperfusion.
    OBJECTIVE: To investigate the effect of different doses of astragalus injection on neuronal apoptosis and expression of caspase-3 in hippocampus of rats after cerebral ischemia reperfusion.
    METHODS: Eighty-four SD rats were randomly divided into three groups: the sham operation group, model group and astragalus injection group. The astragalus injection group was then subdivided into five subsets, according to the astragalus injection doses of 2, 4, 6, 8 and 10 mL/kg. The four-vessel occlusion method was used to build a rat model of cerebral ischemia reperfusion. Terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling staining was used to measure neuronal apoptosis in the hippocampus and Western blot was used to measure the expression of caspase-3 protein.
    RESULTS AND CONCLUSION: Compared with the sham operation group, the apoptosis index and expression of caspase-3 protein increased in the model group (P < 0.05). Compared with the model group, the apoptosis index and expression of caspase-3 protein decreased in the astragalus 4, 6, 8 and 10 mL/kg groups (P < 0.05), but there was no difference in the astragalus 2 mL/kg group (P > 0.05). Compared with the 4 mL/kg group, the apoptosis index and expression of caspase-3 protein decreased in the astragalus 6, 8 and 10 mL/kg groups (P < 0.05) in a dose-dependent manner, but there was no difference among the later three groups (P > 0.05). Astragalus injection can decrease neuronal apoptosis and expression of caspase-3 protein in rat hippocampus after cerebral ischemia reperfusion.

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    Effect of ginkgo biloba extract on activities of primary cells from embryonic rat mesencephalon and dopaminergic neurons  
    Xi Shu-fang1, Wang Qun-jiang2, Zhu Can-hong1, Hu Rong1, Duan Li-rong3
    2012, 16 (20):  3751-3754.  doi: 10.3969/j.issn.1673-8225.2012.20.032
    Abstract ( 268 )   PDF (312KB) ( 416 )   Save

    BACKGROUND: Ginkgo biloba extract (EGB) has the pharmacological activities of free radical and lipid peroxidation scavenging and release of excitatory neurotransmitter resisting.
    OBJECTIVE: To explore whether EGB761 has protective effect on primary cells from embryonic rat mesencephalon and dopaminergic neurons from injuries induced by 1-methyl-4-phenylpyridinum (MPP+) and L-arginine (L-Arg).  
    METHODS: Midbrain nerve cells from Sprague Dawley embryonic rats were isolated and cultured. The cells were divided into control group, MPP+ group, L-Arg group, MPP++L-Arg group, MPP++EGB761 group and MPP++EGB761+L-Arg group.
    RESULTS AND CONCLUSION: Comparison results of cell A values and numbers of tyrosine hydroxylase-positive cells among the groups: these indexes in the MPP+ group and MPP++L-Arg group were significantly decreased compared with the control group, L-Arg group, MPP++EGB761 group and MPP++EGB761+L-Arg group (P < 0.05), those of the MPP++L-Arg group were lower compared with the MPP+group (P < 0.05), and those of the MPP++EGB761+L-Arg group were lower than the MPP++ EGB761 group (P < 0.05). Cell nitric oxide levels: the levels in the MPP+ group and MPP++L-Arg group were obviously higher compared with the control group, L-Arg group, MPP++EGB761 group and MPP++EGB761 L-Arg group (P < 0.05), the level of the MPP++L-Arg group was higher compared with the MPP+ group (P < 0.05), and that of the MPP++EGB761+L-Arg group was higher than the MPP++EGB761 group (P < 0.05). It is indicated that L-Arg can increase the MPP+-induced damage to nopaminergic neurons, and EGB761 has the protective effect on injured primary cells from embryonic rat mesencephalon which may derived from reduction of nitric oxide by inhibiting the activity of inducible nitric oxide synthase.

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    Expression of exogenous Rb gene mediated by piggyBac transposon in retinoblastoma cells   
    Pan Xue-ke1, Chen Zhao1, Tian Si-jia1, Joseph M. Kaminski2○, Yu Ke-ming1, Zhuang Jing1
    2012, 16 (20):  3755-3758.  doi: 10.3969/j.issn.1673-8225.2012.20.033
    Abstract ( 469 )   PDF (487KB) ( 375 )   Save
    BACKGROUND: The introduction of exogenous Rb gene into retinoblastoma is an effective way to inhibit growth of the tumor. piggyBac transposon as a vector is safe and highly efficient, which is a potential vector on gene therapy.
    OBJECTIVE: To explore the transfection efficiency and its effect on inhibiting retinoblastoma by transfecting human retinoblastoma cell line SO-RB50 with an exogenous Rb gene expression system mediated by piggyBac transposon.
    METHODS: pPiggyBac-Rb plasmid that contains a piggyBac transposon sequence and express normal RB gene was constructed and used to transfect SO-RB50 cells alone or with pPiggyBac-helper. Coomassie brilliant blue staining, quantitative real-time PCR and immunofluorescence were adopted to evaluate the transfect efficiency of exogenous Rb gene mediated by piggyBac transposon. The viability of retinoblastoma cells was determined by 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide.
    RESULTS AND CONCLUSION: Transfection efficiency was the highest with help of piggyBac transposon. piggyBac transposon-mediated exogenous Rb gene integrated into genome and obtained a long-term and stable expression.It also had a remarkable inhibition effect on the viability of SO-RB50. All these indicate that piggyBac transposon is a potential vector for retinoblastoma gene therapy.
     
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    pAAV-hSOX9-IRES-tdTomato recombinant plasmid constructs adeno-related virus package★
    Diao Ze-zheng, Yan Guo-qing, Zhang Zhi-wei, Fang Jing, Xu Peng, Xi Yong-ming, Ren Shan, Liu Yong-jun, Sui Ai-hua
    2012, 16 (20):  3759-3762.  doi: 10.3969/j.issn.1673-8225.2012.20.034
    Abstract ( 501 )   PDF (313KB) ( 619 )   Save

    BACKGROUND: As the preliminary experiment for gene therapy in intervertebral disc degeneration, this study aims to construct a recombinant plasmid containing fluorescent pAAV-hSOX9-IRES-tdTomato for adeno-associated virus packaging, in a broader attempt to lay the foundation for late experiments in vitro and in vivo.
    OBJECTIVE: To construct human SOX9 gene overexpressing adeno-associated virus, pAAV-hSOX9-IRES-tdTomato, packaging.
    METHODS: The plasmid pAAV-IRES-tdTomato and plasmid pUC57-hSOX9 were connected into pAAV-hSOX9-IRES-tdTomato by enzyme digestion method. The adeno-associated virus was packaged with plasmid co-transfections method. The recombinant pAAV-hSOX9-IRES-tdTomato was transfected into 293AAV cell by calcium phosphate transfection. The purification and drop of adeno-associated virus was tested by determination of biological titer.
    RESULTS AND CONCLUSION: The results of BLAST sequence comparison analysis showed that, pAAV-hSOX9-IRES-tdTomato exactly matched the synthetic gene sequence hSOX9. The titer is 1×107 TU/mL. Human gene SOX9 recombinant adenoviruses, pAAV-hSOX9-IRES-tdTomato, have been constructed successfully.

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    Construction and identification of RNA interference lentiviral vector for Rac1 gene  
    Wu Yuan1, 2, Zhang Zhuang1, 2, Pan Jian1
    2012, 16 (20):  3763-3767.  doi: 10.396 9/j.issn.1673-8225.2012.20.035
    Abstract ( 303 )   PDF (434KB) ( 284 )   Save

    BACKGROUND: Rac1, which is expressed in a variety of tumors, regulates the invasion and proliferation of cancer cells through signal transduction.
    OBJECTIVE: To construct a RNA interference lentiviral vector that is capable to knock down Rac1 gene, and to detect its interference efficiency.
    METHODS: The gene expression of Rac1 in tongue cancer cells was detected by western blot. The Oligo DNA containing target sequence was synthesized according to the previously confirmed effective sequence of small interfering RNA targeting Rac1 gene. Double-stranded DNA was constructed after annealing, and was cloned into the pMagic4.1 vector after digest with AgeI/EcoRI to construct a lentiviral vector which express short hairpin RNA. Positive clones were identified using PCR. The plasmids and packaging plasmids were cotransfected into 293T cells. The small interfering RNA expression cassettes were generated by real-time PCR; the successfully constructed plasmids were packaged by lentivirus and then transfected into Tca8113 cell. The interference effect of Rac1 gene expression was assayed by fluorescence quantitative PCR and western blot to explore its biological activity.
    RESULTS AND CONCLUSION: RAC1 interference vector was constructed successfully. Western blot results showed that the protein expression of Rac1 was decreased significantly; Real-time PCR results showed that the inhibition rate of pLVT447 was up to 70%. A lentivirus-mediated RNAi vector containing Rac1 gene was successfully constructed, which provides effective small interfering RNA target sequence for the application of RNA interference in the targeting gene therapy of tongue cancer using Rac1.

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    Human apM1 gene transfers into the epicardial adipose tissue to inhibit the formation of coronary atherosclerosis in rabbits
    Yang Wen-kai1, 2, Dong Nian-guo1, Chen Qing3, Wang Xian-guo1, Xie Ting1, Chen Xin-zhong1
    2012, 16 (20):  3768-3772.  doi: 10.3969/j.issn.1673-8225.2012.20.036
    Abstract ( 474 )   PDF (379KB) ( 339 )   Save

    BACKGROUND: The decreased level of adiponectin secreted from the epicardial adipose tissue may be one of the main reasons leading to coronary atherosclerosis.
    OBJECTIVE: To investigate the effect of human apM1 gene transferring into the epicardial adipose tissue on the formation of coronary atherosclerosis in high-cholesterol fed rabbits.
    METHODS: 50 μL mixture of pEGFP-apM1 and liposome was injected into the rabbit pericardial cavity. The epicardial adipose tissue was harvested at 2, 7 and 28 days respectively after injection and was made into fast frozen sections. The transfection efficiency and expression of apM1 were detected by fluorescence microscopy and immunohistochemical method. Twenty-four male New Zealand White rabbits were randomly divided into four groups: normal control group, high-fat group, high-fat plus pEGFP injection group, high-fat plus pEGFP-apM1 injection group. At 4 weeks after injection, the expression of apM1 in epicardial adipose tissue was detected by Western blot analysis.
    RESULTS AND CONCLUSION: In the rabbit epicardial adipose tissue, the high expression of apM1 was detected at 2 days after injection and until to 28 days. The effective transfection was not detected in the epicardial connective and myocardial tissues. The levels of adiponectin, tumor necrosis factor-α, total cholesterol, high density lipoprotein cholesterol and low density lipoprotein cholesterol in the peripheral blood changed insignificantly change after pEGFP-apM1 injection into the pericardial cavity. In high-fat plus pEGFP-apM1 injection group, the distinct increase of adiponectin and decrease of tumor necrosis factor-α (P < 0.01) were measured in the pericardial cavity fluid. Compared with the high-fat group, pEGFP-apM1 injection could decrease the intima/media ratio of left coronary artery to 40.66%. The apM1 gene could be effectively transferred into the epicardial adipose tissue through the pericardial cavity injection. The increase of adiponectin expression in the epicardial adipose tissue could inhibit local inflammatory cytokines release induced by high-fat feeding and inhibit the formation of coronary atherosclerosis.

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    Effect of mutant huntingtin on the expression levels and histone acetyltransferase activity of CREB-binding protein  
    Cong Shu-yan, Zhang Wei, Wang Ya, Shao Hua, Feng Juan
    2012, 16 (20):  3773-3778.  doi: 10.3969/j.issn.1673-8225.2012.20.037
    Abstract ( 384 )   PDF (468KB) ( 497 )   Save

    BACKGROUND: Studies addressing the role of CREB-binding protein (CBP) in the pathogenesis of Huntington's disease are mainly concentrated in the inhibitory effect of aggregate formation on CBP.
    OBJECTIVE: To investigate the effect of mutant huntingtin (htt) on the histone acetyltransferase (HAT) activity and expression levels of CBP in the cell model of Huntington’s disease.
    METHODS: Neuronal phaeochromocytoma rat PC12 cells, stably inducible for GFP-tagged HD exon 1 with either 23 (normal) or 74 (expanded) glutamines (N-htt-Q23 or -Q74), driven by a doxycycline (dox)-dependent Tet-On promoter were used. N-htt expression was observed via immunofluorescence microscopy. The HAT activities of endogenous CBP were determined via a histone H4 acetylation assay with immunoprecipitated CBP. CBP protein levels were observed via Western blot. The effect of specific proteasome inhibitor on CBP protein level was investigated by adding MG-132. CBP mRNA levels were detected by Lightcycler PCR.
    RESULTS AND CONCLUSION: Cells expressing N-htt-Q74 started to form visible aggregates 1 day after dox induction in a small percentage of cells (< 1%), while after 6 days aggregates were present in about 90% of cells. In contrast, N-htt-Q23 distributed evenly throughout the cells and did not form aggregates. Expression of soluble N-htt-Q74 already inhibited the HAT activity of CBP, the inhibitory effect was exacerbated by aggregate formation of N-htt-Q74, while N-htt-Q23 did not affect CBP HAT activity. Expression of soluble N-htt-Q74 already reduced CBP protein levels; the level of CBP was dramatically decreased after 6 days, while N-htt-Q23 did not affect the levels of CBP protein. Neither N-htt-Q74 nor -Q23 affected CBP mRNA levels. The proteasome inhibitor MG-132 partly restored CBP protein levels by affecting protein degradation in the cells expressing N-htt-Q74. The results indicated that decrement of CBP HAT activity and its protein level play important roles in the molecular pathogenesis HD. Aside from aggregated mutant N-htt, soluble mutant N-htt already represses CBP HAT activity and CBP level, providing theoretical evidence for administration of CBP and histone deacetylase inhibitor in the treatment of Huntington's disease in the early stages.

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    Meta-analysis on CDKAL1 gene rs7754860 single neucleotide polymorphisms and type 2 diabetes mellitus susceptibility  
    Wang Juan1, 2, Chen Li-ming3, Sun Hong-xi1, Guo Jun1, Wen Juan-juan1
    2012, 16 (20):  3779-3783.  doi: 10.3969/j.issn.1673-8225.2012.20.038
    Abstract ( 337 )   PDF (279KB) ( 392 )   Save

    BACKGROUND: The pathogenesis of type 2 diabetes is unclear, and genetic studies have shown that there may be genetic predisposition, but the results are not consistent with each other.
    OBJECTIVE: To evaluate the relationship between cyclin-dependent kinase 5 regulatory subunit-associated protein 1-like 1 (CDKAL1) gene rs7754860 G<C single neucleotide polymorphisms and type 2 diabetes mellitus susceptibility.
    METHODS: By searching EMBSE (1974/2011-05), PubMed (1966/2011-05), CNKI (1994/2011-05) and CBM (1978/2011-05), we collected studies about CDKAL1 gene rs7754860 G<C single neucleotide polymorphisms and type 2 diabetes mellitus susceptibility. The odds ratios (OR) of variant allele of CDKAL1 gene rs7754860 was calculated by using Review Manager 5.0 software.
    RESULTS AND CONCLUSION: A total of 19 03 cases and 22 560 controls were included in nine case-control studies. The risk for type 2 diabetes mellitus was higher in the CC group compared with GG group (OR=1.51, 95% confidence interval (CI): 1.24-1.84, P < 0.01); For the dominant genetic model, the risk for type 2 diabetes mellitus had no significant difference between the CC+GG group and GG group (OR=1.22, 95%CI: 0.95-1.56, P=0.12). For the recessive genetic model, the risk for type 2 diabetes mellitus in the CC group was significantly increased as compared with the GC+GG group (OR=1.61, 95%CI: 1.41-1.83, P < 0.01). CDKAL1 gene rs7754860 C<G single neucleotide polymorphisms has close relationship with type 2 diabetes mellitus susceptibility. People with C allele are susceptible to type 2 diabetes mellitus.

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    Monitoring and application of physiological load intensity in exercise   
    Tang Jian, Li Min-hua
    2012, 16 (20):  3784-3788.  doi: 10.3969/j.issn.1673-8225.2012.20.039
    Abstract ( 356 )   PDF (421KB) ( 632 )   Save

    BACKGROUND: Exercise intensity includes physical load intensity, physiological load intensity and subjective feeling intensity. Physiological load intensity as a reflection to exercise intensity can effectively reflect individual adaptation to exercise intensity.
    OBJECTIVE: To elaborate the application progress of heart rate and lactate concentration as physiological load intensity indexes.
    METHODS: A computer-based online search of PubMed and CNKI databases were performed for papers published from 1990 to 2011. Papers related to exercise intensity monitored by physiological load intensity, especially the application of heart rate and lactate as monitoring indexes were included.
    RESULTS AND CONCLUSION: Totally 47 literatures were chosen, which proved that heart rate and lactate as physiological load intensity indexes were effective and widely used in exercise to monitor exercise intensity. The monitoring of individual’s physiological load intensity should be combined with the ultimate value of individual’s physiological load intensity, which could not only obviously improve individual’s training level, but also avoid body injury caused by longtime excessive training. Therefore, the combination of physiological load intensity indexes as well as application conditions and ranges should be considered when exercise intensity is monitored by physiological load intensity. 

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    Molecules and ligands of Wnt signaling pathway involved in the transformation between osteoblasts and osteoclasts
    Wang Hui, Chen Xiao-dong
    2012, 16 (20):  3789-3792.  doi: 10.3969/j.issn.1673-8225.2012.20.040
    Abstract ( 256 )   PDF (387KB) ( 424 )   Save

    BACKGROUND: Wnt signaling pathway regulates the balance of chondrocytes and osteoblasts, chondrocyte proliferation, ossification and degeneration in the degenerative changes of articular cartilage. It has been paid more and more attention.
    OBJECTIVE: To summarize the role of Wnt signaling pathway in osteoarthritis.
    METHODS: A computer-based online search of Wanfang and PubMed databases was performed for related articles published between 2001 and 2011, using key words of “Wnt, pathway, chondrocyte” in Chinese and English, respectively. A total of 24 articles in Wanfang database and 82 articles of PubMed database were collected, and 25 of them were retained according to inclusion criteria.
    RESULTS AND CONCLUSION: A series of proteins and their ligands in secretory Wnt family play an important role in chondrocyte ossification, osteoblast phenotype maintenance and the suppression of bone mesenchymal stem cells pimelosis. The classic Wnt/β-actnin signaling pathway participates in the regulation of osteoarthritis development by activating a series of signal molecules and inhibiting the cross and cascade reactions with other signaling pathways. Meanwhile, the molecules and their ligands in the Wnt signaling pathway are involved in the regulation of chondrocytes and osteoblasts balance, hyperosteogeny and bone reconstruction process.

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    Platelet lysate’s advances in bone tissue engineering 
    Chen Lu, Li Qi-jia, Song Hui-ping
    2012, 16 (20):  3793-3796.  doi: 10.3969/j.issn.1673-8225.2012.20.041
    Abstract ( 259 )   PDF (374KB) ( 430 )   Save
    BACKGROUND: Platelet lysates to repair bone defects cannot only remove the residual cell structure, reduce immunogenicity, but also retain many growth factors to create the conditions for the allograft or xenograft.
    OBJECTIVE: To review the studies that platelets can promote the growth of bone lysis as well as application methods in relevant research.
    METHODS: PubMed (2004-01/2011-01) and CBM databases (2004-01/2011-01) were searched by using the keywords of “tissue engineering, bone defect, gene therapy, growth factor” in English and “platelet plasma, bone tissue engineering, platelet lysate, growth factor” in Chinese, respectively. Studies addressing platelet lysate to promote bone regeneration and repair were included, and repetitive studies were excluded.
    RESULTS AND CONCLUSION: Totally 340 articles were retrieved, and only 23 articles met the inclusive criteria. Up to now, studies have demonstrated that platelet lysate can promote the proliferation of bone marrow mesenchymal stem cells and osteoblasts. Platelet lysate combined with biological ceramic or autogenous bone can significantly promote the repair of bone defects. Rational use of platelet lysate in bone regeneration and repair is of great significance.
    Chen L, Li QJ, Song HP. Effects of brain microvascular endothelial cells and astrocytes following treatment with baicalin on neural stem cell differentiation.Zhongguo Zuzhi Gongcheng Yanjiu. 2012;16(20): 3793-3796.    
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    Application of adenovirus mediated bone morphogenetic protein gene enhanced tissue engineering in bone tissue engineering  
    Pan Le1, Kang Jian-min2
    2012, 16 (20):  3797-3800.  doi: 10.3969/j.issn.1673-8225.2012.20.042
    Abstract ( 271 )   PDF (347KB) ( 247 )   Save

    BACKGROUND: The vector tool is needed when the target gene transduces into target cells, the vector can help the gene transduces into the cells and promote the protein expression. Vector selection is the key of the gene transduction.
    OBJECTIVE: To review research progress of the application of adenovirus medicated bone morphogenetic protein gene enhanced tissue engineering in osteogenic induction and repair of bone defects.
    METHODS: A computer-based search was performed on the PubMed database and CNKI database from January 1989 to January 2012 for the articles on the osteogenic induction of adenovirus medicated bone morphogenetic protein gene transfection and the articles on bone morphogenetic protein gene modified tissue engineered bone repair the bone defects. The key words were “adenovirus, bone morphogenetic protein, gene transfection, bone tissue engineering” in English and Chinese. The repetitive articles and the articles not in English or Chinese were eliminated, and finally, a total of 24 articles were included to review.
    RESULTS AND CONCLUSION: Adenovirus-mediated bone morphogenetic protein gene enhanced tissue engineering transfers the gene encoding with specific function factor to the seed cells using genetic engineering technology, which make the gene to produce the growth factor continuously. Transgenic cells stay on the surface of the stent and release the growth factor slowly, and can promote the proliferation and differentiation of osteoblast precursor cells and induce the directed differentiation of the osteoblast precursor cells in the bone defect area to osteoblasts, in order to repair the bone defects.   

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