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12 November 2011, Volume 15 Issue 46 Previous Issue    Next Issue

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Effects of serum on tumor necrosis factor alpha induced chondrocyte apoptosis following electro-acupuncture Wu Ming-xia, Li Xi-hai, Wu Guang-wen, Li Li, Chen Wen-lie, Liu Xian-xiang 2011, 15 (46):  8551-8555.  doi: 10.3969/j.issn.1673-8225.2011.46.001 Abstract ( 223 )   PDF (1441KB) ( 413 )   Save BACKGROUND: Electro-acupuncture (EA) can promote blood circulation, remove blood stasis, regulate vital energy circulation to relieve pain, relieve rigidity of muscles and activate collaterals, and effectively relieve symptoms of osteoarthritis. OBJECTIVE: To observe the effect of serum on tumor necrosis factor α (TNF-α) induced chontroctye apoptosis following EA. METHODS: Knee joint chondrocytes from 3-week-old SD rats were isolated using enzymatic digestion and apoptosis induced with 10 μg/L TNF-α in vitro. The normal group rats were routinely raised and did not receive any treatment. The osteoarthritis rats were routinely raised, intraarticularly injected with sodium hyaluronate (0.4 μL/g, once a week, for successive 5 weeks), or given EA at the Neixiyan and Waixiyan ( 5, 15 and 30 min/d), and all rats were interfered with serum, and then the apoptotic rate of the chondrocytes was determined. RESULTS AND CONCLUSION: After 15 and 30 minutes of EA at the Neixiyan and Waixiyan, osteoarthritis rats were interfered with serum for 24 hours, the activity of chondrocytes was significantly increased and the apoptotic rate of the chondrocytes was significantly decreased. These findings suggest that serum can inhibit TNF-α induced chondrocyte apoptosis following EA. Related Articles | Metrics

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Effects of Roubin-manipulation on apoptosis of chondrocytes and expression of Bcl-2, Bax and Fas in rabbit knee joint Dai Qi-yi, Ruan Ping, Cui Wei, Li Qiang, Qin Xue-liu, Lin Guang-qi, Liu Jing, Rong Xiang-bin, Han Jie, Qin Jie, Yuan Jing-yang, Wu Zhao-pei, Wang Xiong 2011, 15 (46):  8556-8560.  doi: 10.3969/j.issn.1673-8225.2011.46.002 Abstract ( 328 )   PDF (1475KB) ( 483 )   Save BACKGROUND: Modern studies have demonstrated that excessive cellular apoptosis accelerates the degeneration of cartilaginous tissue. However, effects of mechanical stimulation of manipulation on joint cartilage at the molecular level have been rarely reported. OBJECTIVE: To observe the effects of Roubin-mamipulation on apoptosis of chondrocytes and expression of Bcl-2, Bax and Fas in rabbit knee joint. METHODS: Fifty New Zealand rabbits were randomly divided into normal, sham operation, model, manipulation, and sodium hyaluronate groups. Rabbits in the latter three groups were established into high intraosseous pressure experiment model of right leg by ligating the vena femoralis and gluteae inferiors. One week later, the manipulation group received 10-minute Roubin-manipulation, once every other day, a total of 17 times within 5 weeks. The sodium hyaluronate group received intra-articular injection of 0.6 mL sodium hyaluronate, once a week, for a total of 5 times. RESULTS AND CONCLUSION: At the end of the 8th week, cartilaginous tissue of medial tibial plateau of rabbit right knee joint was harvested. Hematoxylin-eosin staining showed that compared with normal group, cartilaginous tissue degeneration was obvious, and chondrocyte apoptosis rate was significantly increased (P < 0.01), and Bcl-2, Bax and Fas expression in the tibial cartilaginous tissue was significantly increased (P < 0.01) in the model group. Compared with the model group, the cartilaginous tissue degeneration was slight, chondrocyte apoptosis rate and Bax and Fas expression were significantly decreased (P < 0.01), but Bcl-2 expression in the tibial cartilaginous tissue was significantly increased (P < 0.01) in the manipulation and sodium hyaluronate groups. Roubin-mamipulation, like intra-articular injection of sodium hyaluronate, can obviously decrease the apoptosis rate of chondrocytes in rabbit knee joint and the underlying mechanisms are related to upregulated Bcl-2 expression and downregulated Bax and Fas expression. Related Articles | Metrics

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Acute effect of radiofrequency energy on cartilage injury Chen Qi, Wang Da-ping, Zhu Wei-min, Liu Jian-quan 2011, 15 (46):  8561-8564.  doi: 10.3969/j.issn.1673-8225.2011.46.003 Abstract ( 306 )   PDF (1382KB) ( 308 )   Save BACKGROUND: Articular cartilage injury has recently been treated with radiofrequency energy. But there still exists controversy about the energy settings of radiofrequency energy in clinical use. OBJECTIVE: To evaluate the acute effect of bipolar radiofrequency energy with varying intensities on grade Ⅱ cartilage injury, and to explore the relationship between the intensity of radiofrequency energy and curative effect.  METHODS: Three fresh bovine joints were selected to establish grade Ⅱ cartilage injury models. Then the joints were treated by the bipolar radiofrequency energy instrument for 30 seconds under different energy settings (30 W, 50 W, 70 W, 90 W, 110 W). At the same time, the control groups received the corresponding treatment. RESULTS AND CONCLUSION: According to scanning electron microscope, the cartilage surface became smooth after treatment. Therapeutic effect was achieved under the radiofrequency energy setting of 70 W. There was a negative correlation between glycosaminoglycan release rate and radiofrequency energy intensity (P < 0.05). It indicates that cartilage cell activity decreased with the increasing of radiofrequency energy intensity. Therefore, the radiofrequency energy intensity should be controlled in minimum amount without compromising therapeutic efficacy. Related Articles | Metrics

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Effects of liuwei dihuang wan-containing serum on proteoglycan content in cartilage endplates Xu Wu-ji, Li Yue, Wu Guan-bao, Yuan Chao 2011, 15 (46):  8565-8568.  doi: 10.3969/j.issn.1673-8225.2011.46.004 Abstract ( 274 )   PDF (1313KB) ( 427 )   Save BACKGROUND: Integrated cartilage endplate structure maintains its normal physiological function and then affects the physical and pathological conditions of the entire intervertebral disc. Proteoglycan is the main component of cartilage endplate. OBJECTIVE: To observe the effects of liuwei dihuang wan (LWDHW)-containing serum on proteoglycan content in cartilage endplates of tumor necorsis factor α (TNF-α)-induced injury rabbits. METHODS: Intervertebral discs with cartilage endplate were harvested from T12-L2 of 12 rabbits and then were randomly divided into four groups: control 1st day, control 14th day, TNF-α, and TNF-α + LWDHW-containing serum. In the TNF-α group, 5 μg/L TNF-α was added. In the TNF-α + LWDHW-containing serum group, 5 μg/L TNF-α and 10% LWDHW-containing serum were added. Then a 14-day culture was performed. RESULTS AND CONCLUSION: With prolongation of culture time, the number of cartilage endplates was decreased, cellular structure was obviously changed, and collagen content in extracellular matrix was decreased. With addition of TNF-α, collagen content decreased more faster and some crannies appeared. LWDHW-containing serum could delay this process. With prolongation of culture time, the ultrastructure of cartilage endplate was gradually destroyed, TNF-α could aggravate the damage, and LWDHW-containing serum could minimize the damage. With prolongation of culture time, proteoglycan content, chondroitin sulfate, keratan sulfate and hyaluronic acid levels in the cartilage endplates were declined in each group. The decline in the TNF-α group was most obvious (compared to the control group in 14th day, P < 0.05), and LWDHW-containing serum could postpone the decline of biochemical component levels (compared to TNF-α group, P < 0.05). These findings suggest that LWDHW-containing serum can protect the ultrastructure of cartilage endplate and stabilize the levels of biochemical components, thereby exhibiting protective effects on cartilage endplate of intervertebral discs. Related Articles | Metrics

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Primary culture and purification of rat nucleus pulposus cells by anchorage velocity-dependent separation method Sun Hao-lin, Li Chun-de 2011, 15 (46):  8569-8573.  doi: 10.3969/j.issn.1673-8225.2011.46.005 Abstract ( 425 )   PDF (1765KB) ( 630 )   Save BACKGROUND: Nucleus pulposus cells have the problem of phenotype loss during in vitro culture, such as the decreasing expression of type II collagen, Aggrecan and Sox-9, leading to conversion of cartilage-like cells to fibroblast-like cells. OBJECTIVE: To primarily culture rat nucleus pulposus cells purified by anchorage velocity-dependent separation method. METHODS: The lumbar nucleus pulposus from Wistar rats were digested by trypsin and type II collagenase. The first and second passages of cells were isolated and purified by anchorage velocity-dependent separation method during subculture. RESULTS AND CONCLUSION: The third passage of nucleus pulposus cells which were isolated and purified by anchorage velocity-dependent separation method appeared round or multi-angular with activity. Hematoxylin-eosin staining appeared homogeneous with blue nucleus and pink cytoplasm. The positive rate of type II collagen immunohistochemistry staining was 97% and rate of aggrecan immunohistochemistry staining was 95%. Transmission electron microscope results showed that there were many endoplasmic reaticulums, Golgi complexes, free ribosemes and few multilamellar bodies, but no chondriosome. The cell growth curve showed that after subculture, the third passage 3 of nucleus pulposus cells experienced 2 days of latent phase, 3 days of increased logarithmic phase, then went to plateau phase. Anchorage velocity-dependent separation method is a practical and effective method for purifying the primary cultured rat nucleus pulposus cells. The third passage of cells harvested through this method is active and homogeneous. Related Articles | Metrics

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Regulatory effects of Tougu Xiaotong granule on wnt/β-catenin signal pathway of articular chondrocytes Liu Bo-ling, Zou Ji-lin, Liang Gui-qing, Liu Xian-xiang, Li Xi-hai, Wu Guang-wen 2011, 15 (46):  8574-8578.  doi: 10.3969/j.issn.1673-8225.2011.46.006 Abstract ( 449 )   PDF (860KB) ( 561 )   Save BACKGROUND: Tougu Xiaotong granule composed of mornda root, radix paeoniae alba from Hangzhou of China, herba sarcandrae, and szechwan lovage rhizome can effectively delay alteration of macroscopic form of articular cartilage and degeneration of cartilage matrix and chondrocytes. But the underlying mechanism remains poorly understood. OBJECTIVE: To observe the effects of Tougu Xiaotong granule on Wnt4, glycogen synthase kinase 3β (GSK-3β) and β-catenin levels in wnt/β-catenin signal pathway of chondrocytes.  METHODS: SD rat articular chondrocytes were isolated and induced with human interleukin-1β for degeneration. Passage 2 degenerated chondrocytes were randomly divided into a control group and a Tougu Xiaotong granule group. Alcohol extract product of Tougu Xiaotong granule was added to the Tougu Xiaotong granule group. After 4 and 8 days of culture, Wnt4, GSK-3β and β-catenin levels were detected using RT-PCR and western blot methods. RESULTS AND CONCLUSION: Human interleukin-1β could induce the degeneration of chondrocytes. Wnt4, GSK-3β, β-catenin protein expression was observed in the chondrocytes during the early stage of degeneration. But with time going, Wnt4 and β-catenin protein expression was decreased while GSK-3β expression was increased. After intervention with Tougu Xiaotong granule, these phenomena could be reversed. These findings suggest that Tougu Xiaotong granule can induce chondrocytes to synthesize Wnt4 and β-catenin and inhibit GSK-3β expression. Related Articles | Metrics

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Cryopreservation effect of human osteochondral allografts in vitro Qi Jian-hong, Zhao Jian-li, Zhang Yan-ming, Song-Hong qiang, Ge Fu-zhang, Zhang Ming 2011, 15 (46):  8579-8582.  doi: 10.3969/j.issn.1673-8225.2011.46.007 Abstract ( 352 )   PDF (586KB) ( 444 )   Save BACKGROUND: Cartilage allograft transplantation is one of effective methods for treatment of articular cartilage defects. However, the short preservation time of osteochondral allograft in vitro limits its clinical application and the utilization efficiency. OBJECTIVE: To study the preservation effect of articular cartilage allograft with controlled rate cryopreservation. METHODS: The osteochondral allografts were processed by 10% DMSO and then cooled with a controlled rate freezer at a speed of -1 ℃/min to -4 ℃for 30 minutes, to -10℃ for 30 minutes, to -20 ℃ for 30 minutes, and to -40℃ for 30 minutes. Finally, they were put into a -80 ℃ refrigerator for cryopreservation. RESULTS AND CONCLUSION: After cryopreservation for 1, 3 and 6 months and 1 year, the survival rate and viability of chondrocytes and the proteoglycan level in the superficial, middle and deep layers of osteochondral allograft were decreased with time going (P < 0.05) and as compared with fresh group (P< 0.05). These findings suggest that cryopreservation can effectively prolong the survival time of osteochondral allograft. Related Articles | Metrics

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Identification of human nucleus pulposus cells by detecting CD24 expression using flow cytometry Zhang Li, Ma Xun, Guan Xiao-ming, He Li-ming, Zhao Sheng, Song Wen-hui 2011, 15 (46):  8583-8586.  doi: 10.3969/j.issn.1673-8225.2011.46.008 Abstract ( 399 )   PDF (485KB) ( 479 )   Save BACKGROUND: CD24 is known as its specific expression on the surface of nucleus pulposus (NP) cells. Whether detecting CD24 expression can be used as a method to identify NP cells remains poorly understood. OBJECTIVE: To verify whether flow cytometry (FCM) detection of CD24 expression in NP cells is a fast and convenient method to identify NP cells. METHODS: Human NP cells were isolated and cultured from seven patients who underwent various spinal surgeries via enzyme digestion. Type Ⅱ collagen immunohistochemistry staining was proceeded to the well grown passage 2 NP cells. RT-PCR experiments had been done to measure the mRNA expression of type Ⅱ collagen and SOX9 from the same cell source. Also the positive rates of CD24 were detected by FCM. The correlation among these results from the same sample was analyzed. RESULTS AND CONCLUSION: Type Ⅱ collagen immunohistochemistry staining results of in vitro cultured passage 2 NP cells was positive. RT-PCR and FCM also acquired positive results. The positive rate of CD24 was above 50%. Extracellular matrix typeⅡcollagen and SOX9 expression of NP cells correlated to positive CD24 results. Detecting CD24 expression using FCM can identify NP cells fast and conveniently. Related Articles | Metrics

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Effect of static magnetic field on proliferation and functions of rat osteoblasts Sun Xiang-hua, Cai Yong-qin, Yuan Ai-qin 2011, 15 (46):  8587-8590.  doi: 10.3969/j.issn.1673-8225.2011.46.009 Abstract ( 388 )   PDF (1272KB) ( 558 )   Save BACKGROUND: The effect of magnetic field on osteoblast biology remains poorly understood. OBJECTIVE: To evaluate the effect of the static magnetic field (SMF) with different intensities and exposure time on the proliferation, apoptosis and function of osteoblasts. METHODS: Passage 3-5 Sprague-Dawley rat osteoblasts cultured in vitro were exposed to 0, 5, 22, 86, 135 mT SMF for 8, 12 and 24 hours. Cell proliferation and apoptosis changes and osteocalcin and carboxyterminal propeptide of typeⅠprocollagen (PICP) in the cell supernatant were determined. RESULTS AND CONCLUSION: After exposure to 5, 22, 86, 135 mT SMF for 8, 12 hours, cell proliferation index was significantly higher compared with the control group (P < 0.05), and after exposure to SMF for 24 hours, the cell proliferation index was significantly higher only in the 5 mT SMF group than in the control group (P < 0.05). There was no significant difference in percentage of apoptotic cells after 0, 5, 22, 86, 135 mT SMF exposure at 8, 12 and 24 hours. After exposure to SMF for 8 hours, osteocalcin level in the 22, 86 and 135 mT SMF groups was significantly higher than that in the control group (P < 0.05). After exposure to SMF for 24 hours, osteocalcin level in the 135 mT group was significantly lower than that in the control group  (P < 0.05). After exposure to SMF for 8, 12 hours, PICP level was significantly higher in the 22, 86 mT SMF groups than in the control group (P < 0.05). After exposure to SMF for 24 hours, PICP level was significantly lower in the 135 mT SMF group than in the control group (P < 0.05). These findings suggest that short-time low-intensity magnetic field promotes the proliferation of osteoblasts and the secretion of osseous substance, while high-intensity magnetic field or long-time magnetic field exposure inhibits the proliferation of osteoblasts and the secretion of osseous substance. Related Articles | Metrics

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Effects of hypoxia on proliferation and differentiation of rat osteoblasts Gao Wen-kui, Wang De-yuan, Li Zhi-gang, Yan Zi-qiang, Bai Feng, Wang Wei 2011, 15 (46):  8591-8594.  doi: 10.3969/j.issn.1673-8225.2011.46.010 Abstract ( 306 )   PDF (1129KB) ( 355 )   Save BACKGROUND: Hypoxia influences bone metabolism by many means and exhibits negative effects on bone generation and bone healing. OBJECTIVE: To explore the effects of hypoxia on proliferation and differentiation of rat osteoblasts and investigate the underlying mechanisms. METHODS: Primary rat osteoblasts were isolated from excised calvarial bones of neonatal mice and cultured in vitro. The second passage of osteoblasts were cultured under hypoxic (3% O2) and normoxic (20% O2) conditions. RESULTS AND CONCLUSION: The proliferation levels, alkaline phosphatase activities, osteocalcin contents and the number of calcium nodules in the hypoxic group were significantly lower compared with the normoxic group (P < 0.05 or P < 0.01). It means that hypoxia inhibits the proliferation, differentiation and functions of rat osteoblasts. The mRNA expression of bone morphogenetic protein-2 and Runx2 in osteoblasts cultured in 3% O2 was lower than that cultured in 20% O2 ( P < 0.05 or      P < 0.01). It means that the mRNA expression of Runx2 and bone morphogenetic protein-2 was decreased in hypoxic conditions. These findings suggest that hypoxia can inhibit the proliferation and differentiation of rat osteoblasts through inhibiting Runx2 and BMP-2 mRNA expression in vitro. Related Articles | Metrics

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Effects of Jinwugutong capsule on the expression of interleukin1, 6 in osteoblasts Liu Chun-ying, Zheng Wen-kui, Zu Jin-chi, Zhou Yu-juan, Liu Li, Wang En-jun, Li Shao-chun 2011, 15 (46):  8595-8597.  doi: 10.3969/j.issn.1673-8225.2011.46.011 Abstract ( 449 )   PDF (1143KB) ( 420 )   Save BACKGROUND: Our previous studies have shown that Jinwugutong capsule exhibits obvious effects on postmenopausal osteoporosis. OBJECTIVE: To investigate the effects of Jinwugutong capsule-containing serum on the expression of interleukin (IL)-1 and IL-6 in osteoblasts. METHODS: Bilateral ovaries of female Sprague-Dawley (SD) rats were removed to prepare ovarian serum. Ovariotomized (OVX) rats were intragastrically administered Jinwugutong capsule to prepare ovarian medicated serum. Passage 3 osteoblasts which were harvested from rats born within 24 hours were cultured with normal female rat serum (normal serum group), OVX rat serum (OVX serum group) and OVX rat Jinwugutong capsule-containing serum (OVX drug-containing serum group) for 72 hours separately.   RESULTS AND CONCLUSION: ELISA results showed that IL-1 and IL-6 expression in the osteoblasts was significantly increased in the OVX serum group than in the normal serum group (P < 0.01); IL-1 and IL-6 expression in the osteoblasts was significantly decreased in the OVX drug-containing serum group than in the OVX serum group, but there was no significant difference in IL-1 and IL-6 expression between normal serum group and OVX drug-containing serum group. These findings suggest that Jinwugutong capsule can prevent and treat osteoporosis by inhibiting IL-1 and IL-6 expression in osteoblasts.  Related Articles | Metrics

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Effect of adenovirus mediated human RAMP1 gene on proliferation and apoptosis of vascular smooth muscle cells Long Xian-ping, Zhao Ran-zun, Shi Bei, Cui Can, Sheng Jin, Chen Pan-ke 2011, 15 (46):  8598-8602.  doi: 10.3969/j.issn.1673-8225.2011.46.012 Abstract ( 304 )   PDF (1771KB) ( 404 )   Save BACKGROUND: Hyperproliferation of vascular smooth muscle cells (VSMCs) is one of the most important mechanisms of atherosclerosis and restenosis. Receptor activity modifying protein-1 (RAMP1) is receptor subunit of calcitonin gene related peptide (CGRP) and decides the activity of receptor. Exogenous RAMP1 could play an important role in the regulation of VSMCs proliferation.  OBJECTIVE: To investigate the effects of adenovirus mediated human RAMP1 (hRAMP1) gene on proliferation and apoptosis of VSMCs derived from rabbit thoracic aorta. METHODS: VSMCs, isolated from rabbit thoracic aorta, were divided into three groups: hRAMP1 group, empty-adenovirus vector group and control group according to whether transferring Ad-EGFP-hRAMP1 or Ad-EGFP, and each group included four subgroups of 24 hours, 48 hours, 72 hours and 96 hours. RAMP1 protein was detected by immunocytochemistry, and the proliferation of VSMCs was determined by cell counting and MTT assay, and the apoptosis of VSMCs was determined by flow cytometry and TUNEL assay. RESULTS AND CONCLUSION: VSMCs were successfully infected by Ad-EGFP-hRAMP1 or Ad-EGFP, and EGFP expression was increased, peaked at 72 hours and lasted for 96 hours. In the hRAMP1 group, the VSMCs showed hRAMP1 protein expression, and the activity of cell proliferation was significantly inhibited in the hRAMP1 group compared with empty adenovirus vector group and control group (P < 0.05). Also, the level of VSMCs apoptosis was significantly higher in the hRAMP1 group compared with other groups (P < 0.05). These results showed that the over-expression of hRAMP1 mediated by adenovirus vector plays an important role in inhibition of VSMCs proliferation and promotion of VSMCs apoptosis, which may be the therapeutic target of atherosclerosis and restenosis. Related Articles | Metrics

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apelin-13 promotes proliferation of vascular smooth muscle cells by downregulating caveolae expression Mao Xiao-huan, Li Lan-fang, Gao Jing, Yang Li, Liu Wei, Qin Xu-ping, Chen Lin-xi 2011, 15 (46):  8603-8608.  doi: 10.3969/j.issn.1673-8225.2011.46.013 Abstract ( 280 )   PDF (1525KB) ( 645 )   Save BACKGROUND: Previous results have shown that caveolae is likely to participate in apelin-13 promotion of vascular smooth muscle cell proliferation. OBJECTIVE: APJ is a G-protein coupled receptor, and its ligand is apelin peptide. Previously, we reported PI3K and ERK1/2 signal pathway mediated proliferation of vascular smooth muscle cells (VSMCs) induced by apelin-13. This study is to observe caveolae involved in rat VSMCs proliferation induced by apelin-13 and to determine whether caveolin-1 protein binds to signal molecules PI3K, ERK1/2 to mediate VSMC proliferation induced by apelin-13. METHODS: VSMCs were prepared from male Sprague-Dawley rat thoracic aorta by the primary-explant method. The effect of β- cyclodextrin on cell proliferation induced by apelin-13 was measured by MTT assay. VSMCs were incubated with apelin-13 and β-cyclodextrin (β-CD), caveolin-1 expression was detected by western blot assay. Formation of high molecular weight polyprotein component including caveolin-1 and PI3K /ERK1/2 was detected by immunoprecipitation. RESULTS AND CONCLUSION: β-CD (5 mmol/L, 25 hours) significantly enhanced VSMCs proliferation induced by apelin-13   (P< 0.05). Treating VSMCs with apelin-13 (0, 1, 2, 4, 8 µmol/L) downregulated apelin-13-induced expression of caveolin-1 and the effect was distinct at 1µmol/L (P < 0.05). Pretreatment of the cells with 5 mmol/L β-CD could strength the downregulation of apelin-13-induced caveolin-1 expression (P < 0.05). Apelin-13 may induce dissociation of PI3K (ERK1/2) with caveolin-1 in VSMCs compared with the control group. These findings suggest that caveolae is involved in apelin-13-induced VSMCs proliferation. Related Articles | Metrics

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Effective factors for regulating monolayer permeability of human umbilical vein endothelial cells Yan Cheng hui, Yu Hai-bo, Zhang Xiao-lin, Kang Jian, Han Ya-ling 2011, 15 (46):  8609-8612.  doi: 10.3969/j.issn.1673-8225.2011.46.014 Abstract ( 295 )   PDF (1365KB) ( 496 )   Save BACKGROUND: The molecular mechanism underlying exogenous stimulation-caused vascular barrier dysfunction remains poorly understood in the field of vascular pathophysiology. OBJECTIVE: To investigate the effective factors for regulating monolayer permeability of human umbilical vein endothelial cells and search for the effective treatment target. METHODS: Human umbilical vein endothelial cells (HUVECs) were exposed to lipopolysaccharide (LPS) with and without specific inhibitors of RhoA and F-actin stress fibre. Changes in RhoA/SRF/F-actin activity in association with cell permeability and F-actin rearrangement signaling were investigated by immunostaining, transwell assay and western blot analysis. RESULTS AND CONCLUSIONS: When HUVECs were treated with LPS, F-actin cytoskeleton rearrangement was detected by rhodamine-phalloidin staining in a dose and time-dependent manner, meanwhile the HUVEC hyperpermeability was investigated. Furthermore, the RhoA activity and SRF translocalization into nuclei were identified to be associated with elevation of permeability in HUVECs. The effects were blocked when cells were pretreated with Y27632 or Latrunculin B, respectively. Moreover, Latrunculin B added also inhibited the translocalization of SRF into the nuclei mediated by RhoA activity. Our results have identified the RhoA/SRF and their substrates F-actin rearrangement mediated LPS-induced changes in HUVECs permeability. Related Articles | Metrics

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A mouse model of lupus induced by nucleoprotein with the same genetic background Xie Jun, Lin You-kun 2011, 15 (46):  8613-8616.  doi: 10.3969/j.issn.1673-8225.2011.46.015 Abstract ( 322 )   PDF (1053KB) ( 354 )   Save BACKGROUND: A mouse model of spontaneous lupus cannot be used for study of pathogenic factors other than genetic factors. OBJECTIVE: To establish a mouse model of lupus induced by Balb/c mouse nucleoprotein with the same genetic background. METHODS: Thirty Balb/c mice of SPF grade, aged 4-6 weeks, were randomly and evenly divided into three groups. The V1 group mice were intramuscularly administered nucleoprotein extracted from the same strains of Balb/c mice, once every other 3 weeks, for a total of four times. The V2 group mice were administered the same volume of phosphate buffered saline. The V3 group mice served as normal controls. The 24-hour urine protein, serum antinuclear antibody, anti-ds-DNA antibody and direct immunofluorescence of mouse kidney were detected at 3 weeks after the last immunization. RESULTS AND CONCLUSION: The 24-hour urine protein, anti-ds-DNA antibody, and antinuclear antibody in the V1 group were significantly higher than those in the V2 and V3 groups. There was IgG immune complex deposition in the renal glomerulus of V1 group. However, in the V2 and V3 groups, renal glomerulus contour was not observed, and only non-specific fluorescence could be observed. A mouse model of lupus can be successfully induced by immunizing Balb/c mouse with Balb/c mouse nucleoprotein with the same genetic background. Related Articles | Metrics

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Effects of magnetic stimulation on nerve conduction velocity and the expression of growth associated protein 43 in rats after sciatic nerve injury Wang Wei, Yuan Xiu-hua, Wang Zhong-li, Liu Ning, Li Long-guang 2011, 15 (46):  8617-8620.  doi: 10.3969/j.issn.1673-8225.2011.46.016 Abstract ( 393 )   PDF (1397KB) ( 521 )   Save BACKGROUND: Magnetic stimulation may promote the repair of nerve injury. OBJECTIVE: To explore the effect of magnetic stimulation on nerve conduction velocity and growth associated protein 43 expression of corresponding spinal motor neuron in rats after sciatic nerve injury. METHODS: A number of 60 SD rats were randomly assigned into 3 groups: experiment group (n=24), model group (n=24) and sham-operation group (n=12). The mouse sciatic nerve was clamped by a new hemostatic forcep with a length of 17 centimeters. The clamping tension was adjusted by locking the 2nd interlocking teeth with a pressure of 21.95×103 Pa, lasting for 10 seconds to construct sciatic nerve injury model. At 24 hours after the model construction, the experiment mice were treated with a daily magnetic stimulation of 0.09 T. RESULTS AND CONCLUSION: According to immunohistochemical staining, at the 2nd, 4th, 8th and 12th weeks after model construction, the expression of growth associated protein 43 in L4-5 motor neurons in the experiment group was significantly higher than that of the model group at the same time point (P < 0. 05). Electrophysiology examination showed that compare with the model group, the regenerated nerve of the experiment group had faster conduction velocity, higher amplitude and shorter latent period at the 12th week after model construction (P < 0.05). These results demonstrate that the magnetic stimulation can speed up the nerve conduction of injured sciatic nerve, increase the expression of growth associated protein 43 in corresponding motor neurons and promote the repair of sciatic nerve injury in rats. Related Articles | Metrics

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Comparison of growth and metabolism of cholangiocarcinoma cells in two-dimensional and three-dimensional culture systems Li Lei, Zhou Wei-jia, Wang Jian-dong, Ye Zhao-yang, Zhou Yan, Tan Wen-song 2011, 15 (46):  8621-8624.  doi: 10.3969/j.issn.1673-8225.2011.46.017 Abstract ( 478 )   PDF (1280KB) ( 341 )   Save BACKGROUND: Two-dimensional (2D) culture model is a common tool for tumor research and drug screening in vitro; however, there are some limitations. OBJECTIVE: To establish a three-dimensional (3D) culture model of cholangiocarcinoma cells and to compare the difference in cell growth and metabolism between 2D and 3D culture systems. METHODS: Cholangiocarcinoma cell line QBC939 was cultured in 2D and Cytodex-3 or Cytopore-2 microcarrier-based 3D culture systems. Cell morphology was observed under inverted light microscopy. Total cell number, glucose and lactate concentrations were determined, and the kinetic constants of cell growth and metabolism were calculated. RESULTS AND CONCLUSION: In comparison with 2D and Cytodex-3-based 3D culture systems, Cytopore-2-based 3D culture system achieved higher cell adhesion rate, higher cell density, and lower specific glucose consumption and specific lactate production rates (P < 0.05). In addition, cells were uniformly distributed in Cytopore-2 microcarriers. Therefore, Cytodex-3 or Cytopore-2 microcarriers can be used to establish cholangiocarcinoma 3D culture model, and Cytopore-2 3D culture system is better than Cytodex-3 system. Related Articles | Metrics

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Effects of Naoyian serum on transforming growth beta 1 mRNA expression in cultured rat cerebral microvascular endotheal cells with hypoxia Wan Sai-ying, Li Xing-qun, Zhi Yi-hui, Luo Yun 2011, 15 (46):  8625-8629.  doi: 10.3969/j.issn.1673-8225.2011.46.018 Abstract ( 401 )   PDF (1905KB) ( 395 )   Save BACKGROUND: Clinical and animal experiments have confirmed that Naoyian (NYA) serum can promote hematoma absorption during acute cerebral hemorrhage and subsequent neurological function recovery and improve patients’ living quality. OBJECTIVE: To determine the effects of NYA serum on the expression of transforming growth β1 (TGF-β1) mRNA in cultured rat cerebral microvascular endothelial cells (RCMECs) with hypoxia. METHODS: RCMECs were cultured with the culture medium consisting of 5% NYA serum or 5%normal serum in an anaerobic incubator for 18 hours so as to establish the hypoxia models. The cultured RCMECs were randomly divided into four groups: normal, model, normal serum and NYA serum. The viability of RCMECs was determined by MTT method. TGF-β1 mRNA expression was measured by reverse transcription-polymerase chain reaction (RT-PCR).  RESULTS AND CONCLUSION: The cultured RCMECs exhibited damage after 18 hours of hypoxia injury. The number of surviving RCMECs in NYA serum group was significantly higher than that in the model and normal serum groups, as determined by MTT method. TGF-β1mRNA expression was increased by hypoxia in the cerebral microvascular endothelial cells. TGF-β1mRNA expression in the NYA serum group was significantly lower than that in the other groups (P < 0. 05). These findings suggest that NYA serum downregulating TGF-β1mRNA expression may be one of mechanisms by which NYA serum inhibits apoptosis of cerebral microvascular endothelial cells and then protects cerebral microvascular endothelial cells from hypoxia injury. Related Articles | Metrics

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Angiogenesis and expression of fibroblastic growth factor mRNA and protein after needle-embedding therapy of Neiguan in miniature pigs with myocardial ischemia injury Yang Xiao-fang, Cui Jin, Liu Xiao-yu, Xu Zhao, Zhang Xiao-shan, Feng Lin, Wang Xing-gui, Qian Ning 2011, 15 (46):  8630-8634.  doi: 10.3969/j.issn.1673-8225.2011.46.019 Abstract ( 376 )   PDF (1705KB) ( 458 )   Save BACKGROUND: Angiogenesis and fibroblast growth factors are closely related to the repair of myocardial ischemia injury. Myocardial ischemia injury can be prevented and cured by acupuncturing Neiguan. It is unclear that whether the mechanism behind this is related to the angiogenesis and fibroblast growth factors. OBJECTIVE: To explore the effect of needle-embedding therapy of Neiguan on the angiogenesis and expression of fibroblast growth factor gene and protein in miniature pigs after myocardial ischemia injury. METHODS: A number of 32 miniature pigs were randomly divided into 4 groups. The myocardial ischemia injury model was established by ligating the left anterior descending coronary artery. The left anterior descending coronary artery of the sham-operated pigs was only threaded without ligation. Pigs in Neiguan group and Geshu group were treated with needle-embedding therapy of Neiguan and Geshu respectively after establishing the myocardial ischemia injury model. Model group and sham-operated group received no treatment. RESULTS AND CONCLUSION: According to immunohistochemical staining, the capillary density of cardiac muscle in miniature pigs was lowered after coronary artery ligation (P < 0.05). The capillary density of cardiac muscle increased on the 7th day after needle-embedding therapy of Neiguan and Geshu (P < 0.05 or P < 0.01). Neiguan group preceded Geshu group (P < 0.05). According to real time PCR and western blot, the fibroblast growth factor mRNA and protein expression of myocardial tissue in miniature pigs were significantly improved after coronary artery ligation (P < 0.05 or P < 0.01). The fibroblast growth factor mRNA and protein expression of Neiguan group and Geshu group improved, and the effect of needle-embedding therapy of Neiguan was the most evident (P < 0.05). It reveals that needle-embedding therapy of Neiguan and Geshu can increase the fibroblast growth factor mRNA and protein expression of myocardial tissue, leading to an increase of cardiac muscle capillary density, therefore ameliorate the myocardial ischemia. And the needle-embedding therapy of Neiguan is better than that of Geshu. Related Articles | Metrics

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Low-intensity pulsed ultrasound promotes platelet derived growth factor and insulin-like growth factor 1 expression during tibial fracture healing in rats Li Yan-ru, Zhang Chang-jie, Liu Han-liang 2011, 15 (46):  8635-8638.  doi: 10.3969/j.issn.1673-8225.2011.46.020 Abstract ( 318 )   PDF (1257KB) ( 368 )   Save BACKGROUND: Low-intensity pulsed ultrasound has been demonstrated to promote fracture healing. OBJECTIVE: To investigate the effects of low-intensity pulsed ultrasound on platelet derived growth factor and insulin-like growth factor 1 expression during tibial fracture healing in rats. METHODS: Rat models of closed fracture in unilateral tibia were established and then low-intensity pulsed ultrasound treatment was daily performed. RESULTS AND CONCLUSION: At 14 and 21 days after model preparation, bone calcus amount and thickness in rat tibial fracture models treated with low-intensity pulsed ultrasound were significantly increased (P < 0.05) and at 7 and 14 days after model preparation, platelet derived growth factor and insulin-like growth factor 1 expression in bone callus was significantly increased (P  < 0.05). These findings suggest that low-intensity pulsed ultrasound increases platelet derived growth factor and insulin-like growth factor 1 expression during tibial fracture healing in rats, accelerate chondrocytes maturation and endochondral ossification, and thereby promote fracture healing. Related Articles | Metrics

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Early application of growth response-1 small interfering RNA can inhibit the expression of interleukin-1 beat mRNA induced by cigarette smoke in rat aortic vascular smooth muscle cells Liu Xue-lian, Liu Gui-nan, Guo Hong-tao 2011, 15 (46):  8639-8642.  doi: 10.3969/j.issn.1673-8225.2011.46.021 Abstract ( 250 )   PDF (660KB) ( 389 )   Save BACKGROUND: Researches show that the expression of interleukin-1β is elevated in atherosclerosis vascular cells. OBJECTIVE: To investigate the effect of early growth response-1 small interfering RNA on the expression of interleukin-1β mRNA induced by cigarette smoke extract in rat aortic vascular smooth muscle cells. METHODS: Rat aortic vascular smooth muscle cells were cultured in vitro, then treated with cigarette smoke extracts of different volume fractions (0%, 5%, 10%, 20% and 40%) to screen for the optimal volume fraction. On that basis, the cells were treated for 0, 8, 16 and 24 hours to determine the optimal intervention time. In the end, the expression of growth response-1 was shut down by growth response-1 interfering RNA. RESULTS AND CONCLUSION: According to RT-PCR, cigarette smoke extract was able to induce interleukin-1β mRNA expression in a dose-dependent and time-dependent manner in rat aortic vascular smooth muscle cells. The optimal intervention time and volume fraction of cigarette smoke extract were 8 hours and 10%. Results presented that cells transfected with small interfering RNA targeting growth response-1 for 8 hours showed a significant reduction in interleukin-1β mRNA expression (P < 0.05), in response to 10% cigarette smoke extract treatment. It proves that early growth response-1 small interfering RNA can inhibit the expression of interleukin-1β mRNA induced by cigarette smoke in rat aortic vascular smooth muscle cells. Related Articles | Metrics

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Repeated rapid skin expansion promotes the expression of epidermal growth factor and proliferating cell nuclear antigen Ren Gui-yun, Wang Shou-jun, Dong Fu-sheng, Yu Mei-qing, Hao Fu-liang, Zhang Xu-dong 2011, 15 (46):  8643-8647.  doi: 10.3969/j.issn.1673-8225.2011.46.022 Abstract ( 392 )   PDF (808KB) ( 416 )   Save BACKGROUND: Repeated rapid skin expansion has been proposed because of deficiency in conventional expansion and rapid expansion currently used in the clinic. OBJECTIVE: To investigate the effect of the repeated rapid skin expansion on the expression of epidermal growth factor (EGF) and proliferating cell nuclear antigen (PCNA). METHODS: Forty-eight male rabbits were randomly divided into three groups: repeated rapid expansion, rapid expansion, conventional expansion (16 rabbits per group). The samples were collected at different phases and were fixed in 10% formaldehyde solution. The expression of EGF and PCNA was observed with immunohistochemistry. RESULTS AND CONCLUSION: At the 2nd and 3rd weeks, EGF expression was significantly greater in the repeated rapid expansion group than in the rapid expansion group and conventional expansion group (P < 0.05). PCNA positive staining was significantly higher in the repeated rapid expansion group than in the other two groups at each time stage except for the 3rd week (P < 0.05). These findings suggest that repeated rapid skin expansion can promote the proliferation of fibroblasts and epidermal cells by raising the content of EGF in the expanded skin. Related Articles | Metrics

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Effects of body cavity-like temperature unsuitable for in vitro culture on rat type A spermatogonial cells Deng Cheng-liang, Wang Da-li, Li San-hua, Zhu Jing-jing, Tang Xiu-jun, Gong De-zhang 2011, 15 (46):  8648-8652.  doi: 10.3969/j.issn.1673-8225.2011.46.023 Abstract ( 318 )   PDF (743KB) ( 409 )   Save BACKGROUND: Temperature would influence the proliferation and apoptosis of spermatogonial cells cultured in vitro. OBJECTIVE: To investigate the effects of temperature on cell cycle and proliferation-related gene c-kit and cyclinD3 of type A spermatogonial cells cultured in vitro. METHODS: The isolated spermatogenic cells by two-step enzymatic digestion were co-cultured with sertoli cells in an incubator at 32 ℃ and 37 ℃ to observe the morphological changes and growth characteristics every day. On the 8th day of in vitro culture, type A spermatogenic cells were separated by gradient centrifugation and differential adhesion method. RESULTS AND CONCLUSION: On the 1st day of in vitro culture, suspending spermatogenic cells which were round or oval and the adherent sertoli cells that were fusiform or irregular shape were observed in 32 ℃ group and 37 ℃ group. Starting from the 4th day, the number and growth state of spermatogenic cells were gradually decreased in the 37 ℃ group, at the 8th day, the number of spermatogenic cells in the 37 ℃ group was significantly lower than that in the 32 ℃ group (P < 0.05). Flow cytometry results showed that at the 8th day of in vitro culture, the proportion of type A spermatogonial cells at S phase in the 32 ℃ group was significantly higher than that in the 37 ℃ group (P < 0.05); the relative mRNA and protein expression levels of c-kit, cyclinD3 in type A spermatogonial cells were significantly higher in the 32 ℃ group than in the 37 ℃ group (P < 0.05). Results indicate that spermatogenic cells cultured in vitro would inhibit the proliferation of type A spermatogonial cells at body cavity-like temperature (37 ℃). Related Articles | Metrics

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Olfactory ensheathing cell viability in different cryopreservation systems Liu Yu-liang, Wei Kai-bin, Liu Hong, He Yu-qin, Zhuo Feng,Lü Xin-gang 2011, 15 (46):  8653-8656.  doi: 10.3969/j.issn.1673-8225.2011.46.024 Abstract ( 326 )   PDF (308KB) ( 468 )   Save BACKGROUND: A proper preservation method would be of important significance for experiments and clinical application of olfactory ensheathing cells (OECs) OBJECTIVE: To explore proper cyropreservative systems for OECs. METHODS: OECs during the logarithmic growth phase were harvested, cryopreserved for 1, 3 and 6 months and then revitalized. RESULTS AND CONCLUSION: MTT assay and tryplan blue staining showed that cells exhibited highest viability after treatment with 5% dimethyl sulfoxide (DMSO)-6% hydroxyethyl starch (HES), followed by 10% DMSO, and lastly the 5% DMSO. Use of refrigerator or cryogenic control system with different cryopreservation time did not yield obvious effects on viability of OECs. Therefore, 5% DMSO-6%HES is recommended as a cryopreservative agent for OECs. Related Articles | Metrics

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Cultivation of endothelial progenitor cells from human peripheral blood Li Qin-shan, Liu Yang, Li Yan-ju 2011, 15 (46):  8657-8660.  doi: 10.3969/j.issn.1673-8225.2011.46.025 Abstract ( 363 )   PDF (654KB) ( 379 )   Save BACKGROUND: Although peripheral blood in adults can provide abundant source of cells, the content of endothelial progenitor cells is low. It is necessary to establish a stable and mature in vitro cultivation system of endothelial progenitor cells in order to achieve a better application in tissue engineering and cell therapy. OBJECTIVE: To establish a stable cultivation system of endothelial progenitor cells from human peripheral blood in vitro, including cell isolation, culture and expansion. METHODS: Mononuclear cells were isolated from the human peripheral anticoagulant blood sample by density gradient centrifugation, and then the cells were plated on fibronectin-coated culture dishes. The cells were cultured in endothelial progenitor medium. Non-adherent cells were washed away. The adherent cells were collected at the 6th day after cultivation. The morphologic changes of endothelial progenitor cells were observed by hematoxylin-eosin staining through invert microscope. Growth curves of cells of passages 1 and 3 were measured by MTT and cell counting. The cultured cells were identified by flow cytometry based on the presence of specific surface markers in progenitor cells and endothelial cell line. RESULTS AND CONCLUSION: According to the growth curves, cells entered logarthmic growth phase at the 3rd day after inoculation, and then entered platform phase at the 6th day after inoculation. Cell proliferation rate slowed with increasing passage times. The stem cell surface markers CD34, CD133 and endothelial cell surface markers von willebrand factor and vascular endothelial growth factor receptor 2 were expressed at the same time. These results demonstrate that endothelial progenitor cells derived from human peripheral blood can be propagated in vitro. Related Articles | Metrics

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Cloning and sequencing of mature peptiede of human bone morphogenetic protein 4 cDNA Yuan Shao-hui, Liu Wei, Wu Bin-qi, Han Xi-guang, Bi Zheng-gang 2011, 15 (46):  8661-8664.  doi: 10.3969/j.issn.1673-8225.2011.46.026 Abstract ( 235 )   PDF (666KB) ( 373 )   Save BACKGROUND: Studies have shown that bone morphogenetic protein 4 (BMP-4) has strongest osteoinductive capacity, but its content in tissue is low. OBJECTIVE: To clone human BMP-4 (hBMP-4) mature peptide gene.  METHODS: According to the hBMP-4 sequence reported in Genebank, two primers were designed. The gene sequence of hBMP-4 mature peptide was gained by one step RT-PCR from the mRNA, which was extracted from human placenta. Then it was cloned into the vector of PGEM-T and the resultant plasmid was transformed into DH5α.The sequence of the plasmid BMP-4 was detected by DNA sequence machine.  RESULTS AND CONCLUSION: DNA agarose electrophoresis showed that the product of RT-PCR was about 370 bp and it was consistent with the reported hBMP-4 sequence in Genebank. These findings suggest that the mature peptide of hBMP-4 gene can be successfully cloned from human placenta by RT-PCR. Related Articles | Metrics

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Establishment of a method for primary culture of mouse Leydig cells in vitro Zhao Wei-min, Yang Jian-ying, Dai Tao, Zhang Gong-bao, Yuan De-pin 2011, 15 (46):  8665-8667.  doi: 10.3969/j.issn.1673-8225.2011.46.027 Abstract ( 358 )   PDF (514KB) ( 654 )   Save BACKGROUND: Tissue explant culture of Leydig cells leads to difficult control of contamitation and trypsin digestion of Leydig cells may injury cells. OBJECTIVE: To establish a simple and effective method for primary culture of mouse Leydig cells. METHODS: Mature mouse Leydig cells were digested by collagenase digestion and were purified by Percoll gradient centrifugation. Cell viability was identified by Trypan blau staining and cell purity was determined by 3 β-hydroxysteroid dehydrogenase staining. RESULTS AND CONCLUSION: The cell purity was > 90%. The Leydig cells retained complete morphology, raid proliferation and good adherence. Results showed that an in vitro primary culture model of mouse Leydig cells was successfully established. Related Articles | Metrics

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Construction and identification of delta opioid receptor-short hairpin RNA eukaryotic expression vector Hu Song-quan, Wang Peng, Yang Hui 2011, 15 (46):  8668-8670.  doi: 10.3969/j.issn.1673-8225.2011.46.028 Abstract ( 298 )   PDF (512KB) ( 354 )   Save BACKGROUND: Long-term application of opioid receptor for treatment of pains would cause drug tolerance or addiction, which is possibly related to upregulated delta opioid receptor (DOR) density. OBJECTIVE: To design and construct short hairpin RNA (shRNA) eukaryotic expression plasmids targeting DOR gene which may play an important role in morphine tolerance.   METHODS: Three pairs of short chain oligonucleotides targeted to rat DOR mRNA (Accession No: NM_012617) were designed and synthesized individually based on the sequence of rat DOR mRNA at first. Then the synthestic sense and antisense oligonucleotide strands were mixed together in annealing buffer to form 3 DNA duplexes. Subsequently, the DNA segments were cloned into pGenesil-1.2 vectors respectively. At last, the plasmids were identified by restriction analysis and sequencing test. RESULTS AND CONCLUSION: The results of restriction analysis and sequencing test showed that all three designed DOR shRNA-expression duplexes were successfully inserted into the plasmid vector pGenesil-1.2 respectively and recombinant plasmid vectors were constructed meeting our aims. The DOR-shRNA expression vectors were constructed successfully and could be used in RNAi’s study on morphine tolerance. Related Articles | Metrics

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Construction of human glucose regulated protein 78 eukaryotic expression vector and expression in retinal pigment epithelium cells Li Man-li, Qi Hui, Sun Lei, Wu Ya-zhen 2011, 15 (46):  8671-8675.  doi: 10.3969/j.issn.1673-8225.2011.46.029 Abstract ( 349 )   PDF (607KB) ( 352 )   Save BACKGROUND: Glucose regulated protein 78 (grp78) is a molecular chaperone protein, which is mostly located on the endoplasmic reticulum, and protects cells from cell apoptosis. OBJECTIVE: To construct the pEGFP-grp78 fusion protein eukaryotic expression vector and to detect their expression in human retinal pigment epithelium (RPE) cell lines. METHODS: The encoding fragment of human grp78 gene was obtained from a recombinant plasmid POTB7-grp78 using PCR, and it was digested by restriction enzymes Hind Ⅲ and KpnⅠ. Then the purified grp78 gene fragment was inserted into the GFP expression vector PEGFP-N1, and the recombinant plasmid PEGFP-grp78 was identified by PCR analysis and DNA sequencing. The recombinant plasmids were transfected into RPE cells by lipofectamin. RPE cell lines with grp78 stable expression were established by G418 selection. The expression of grp78 gene was detected by RT-PCR. The expression of fusion proteins was assayed by fluorescence microscopy. RESULTS AND CONCLUSION: We successfully constructed the expression vector of pEGFP-grp78 fusion protein and the sequence results were matched to the cDNA of human grp78. Stably transfected RPE cell line was established and grp78 was expressed successfully. Related Articles | Metrics

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Correlation between masticatory muscle ultrasound data and dental arch morphology of young males Yu Ying-zi, Mi Cong-bo, Zhao Ya-ping, Zu Qing, Liang Qi-bin, Lin cheng, Wang Qiao-yun 2011, 15 (46):  8676-8680.  doi: 10.3969/j.issn.1673-8225.2011.46.030 Abstract ( 403 )   PDF (733KB) ( 664 )   Save BACKGROUND: Various studies have shown the association between masticatory muscle morphology and craniofacial morphology. However, the interaction between masticatory muscle morphology and dental arch form remains poorly understood. OBJECTIVE: To investigate the influence of masticatory muscle morphology and function on dental arch forms and the underlying mechanisms. METHODS: A total of 69 males with normal occlusion, aged 23.6 years (mean 18-28 years) were included in this study. Ultrasound scanning was used to measure the perimeter, area, width, length, mean thickness and maximal thickness of masseter muscle and the mean thickness and maximal thickness of anterior temporal muscle. All items were measured under mandibular postural position. The arch forms were measured. Correlation analysis was done between the ultrasound data and dental arch form. RESULTS AND CONCLUSION: There was a positive correlation between the cross-section area of masseter muscle and the length of maxillary antero-middle dental arch, between the maximal thickness of masseter muscle and the width of mandibular middle dental arch and the perimeter of maxillary dental arch (P < 0.05). There was a negative correlation between the thickness of anterior temporal muscle and the length of maxillary anterior and antero-middle dental arch (P < 0.05). The results indicate that the function and morphology of masseter and temporal muscle may be one of the factors influencing the morphology of dental arch. Related Articles | Metrics

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Chymase expression in deep partial-thickness burn wounds after burn injury in hamsters Dong Xiang-lin1, Geng Zhong-li, Gao Wei-cheng, Zhao Yang, Qin Tao, Qiao Xing, Ma Juan, Ma Shao-lin 2011, 15 (46):  8681-8684.  doi: 10.3969/j.issn.1673-8225.2011.46.031 Abstract ( 397 )   PDF (374KB) ( 403 )   Save BACKGROUND: Little is known about mast cell chymase in burn injury in hamsters. OBJECTIVE: To investigate whether mast cell chymase is involved in burn injury and the change of chymase expression in burn wound. METHODS: Deep partial-thickness scald in hamsters was induced with contact time of 12 seconds by 20 mL 75 ℃ water in a   50 mL syringe. Chymase mRNA level and chymase activity were analyzed before burn and at 1, 3, 7, 14 days after burn in deep second-degree scald wound. RESULTS AND CONCLUSION: Quantitative real-time RT-PCR and radioimmunoassay results showed that chymase gene expression and chymase activity in burn wound were significantly increased after burn and were highest at 3 days after burn. Expression of mast cell chymase would be involved in burn injury and changed at different stages after burn in hamsters. Related Articles | Metrics

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Age-related development characteristics of junior isokinetic muscle torque Xue Shu-de, Zhang Xiu-zhen 2011, 15 (46):  8685-8688.  doi: 10.3969/j.issn.1673-8225.2011.46.032 Abstract ( 412 )   PDF (1105KB) ( 387 )   Save BACKGROUND: Balanced development growth of muscle power in junior period plays an important role in human development. OBJECTIVE: To explore the biomechanical rule of muscle power development in junior period. METHODS: Dynamic muscle power test of flexor and extensor muscles of bilateral shoulders and hip was performed in 30 boys and 30 females through the use of Kintech isokinetic system, with a test speed of 60 (°) / s and test times for stretch six times. RESULTS AND CONCLUSION: The relative peak torque of flexor and extensor muscle of hip and shoulder joints in all involved subjects was gradually decreased with the increase of test speed and was gradually increased with the increasing age. Under different test speeds, the relative peak torque of flexor and extensor muscle of hip and shoulder joints was relatively greater in boys than in girls (P < 0.05), but there was no significant difference in relative peak torque of flexor and extensor muscle of hip and shoulder joints between boys and girls. Related Articles | Metrics

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Recombinant human vascular endostatin inhibits hyperplastic scar of rabbit ears Wang Peng, Xue Bin, Jiang Li-zhu, Lei Bin, Song Peng-yu, Ding Yun-peng, Li Jing, Jiang Juan 2011, 15 (46):  8689-8692.  doi: 10.3969/j.issn.1673-8225.2011.46.033 Abstract ( 418 )   PDF (1032KB) ( 640 )   Save BACKGROUND: Current treatments of hyperplastic scar are poor and limited, so it is necessary to seek a new effective drug to treat the hyperplastic scar. OBJECTIVE: To investigate the effect of recombinant human endostatin injection on hyperplastic scars of New Zealand big-eared rabbit ears. METHODS: 16 New Zealand big-eared rabbit were used to establish hyperplastic scar animal models and then the rabbits were randomly divided into an experimental group and a control group with 8 rabbits in each group. After epithelialization of rabbit ear wound, rabbits in the experimental group were injected with recombinant human endostatin, and those in the control group were injected with physiological saline. After 30 days, changes in hyperplastic scar were observed and hematoxylin-eosin staining was performed to investigate the expression of vascular endothelial growth factor in the scar tissue by immunohistochemisty. RESULTS AND CONCLUSION: At 30 days after intervention, the scar tissue color became weaker, texture was softer, and scar thickness was thinner in the experimental group than in the control group. Hematoxylin-eosin staining showed that compared with the control group, dermis was thinner, fibroblasts in unit area were fewer, exhibiting a parallel arrangement, and capillary caliber was smaller in the experimental group. The hyperplastic scar index was significantly lower in the experimental group than in the control group (P < 0.01). Vascular endothelial growth factor positive expression in the experimental group was significantly lower than that in the control group (P < 0.01). These findings suggest that recombinant human endostatin injection maybe a better choice for treating hyperplastic scar by decreasing vascular endothelial growth factor protein expression. Related Articles | Metrics

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Relations of surface electromyogram signal with muscle contraction and motor centers in rabbit gastrocnemius muscle Li Xin, Zhao Hai-hong, Sun Ai-ping, Zhang Xue-min, Cao Xiao, Zhao Wen-ru 2011, 15 (46):  8693-8697.  doi: 10.3969/j.issn.1673-8225.2011.46.034 Abstract ( 350 )   PDF (1674KB) ( 1238 )   Save BACKGROUND: In the previous verification tests, muscle contraction is induced by electric stimulation. Therefore, the detected surface electromyogram signal is mixed with applied electric current. So the relations between surface electromyogram signal, muscle contraction and upper motor centers can not be fully analyzed, especially the latter one. OBJECTIVE: To assess the relations between surface electromyogram signal, muscle contraction and upper or lower motor centers in New Zealand rabbit gastrocnemius muscle. METHODS: The right heel tendon of a conscious rabbit was pricked by an acupuncture needle to induce gastrocnemius muscle contraction. The right sciatic nerve was severed under the local anesthesia. The intensity and category of the gastrocnemius muscle surface electromyogram signal were measured respectively when pricking the right heel tendon, clamping the distal end of the severed sciatic nerve, pricking the right gastrocnemius muscle and inducing passive gastrocnemius muscle movement. Then the spinal cord was cut off at L3 plane under the local anesthesia. The gastrocnemius muscle contraction was induced by pricking the left heel tendon. The intensity and category of the gastrocnemius muscle surface electromygram signal were measured. RESULTS AND CONCLUSION: Under normal condition, a high magnitude (400 μV) and wide bottom (4 seconds) voluntary wave was detected when voluntary gastrocnemius muscle contraction was induced by pricking the heel tendon. After cutting off the spinal cord, a low magnitude ( < 100 μV) and narrow bottom ( < 1 second) reflex wave was detected when reflexive gastrocnemius muscle contraction was induced by pricking the heel tendon. After severing the sciatic nerve, muscle contraction cannot be induced when pricking the heel tendon, the muscle or the sciatic nerve distal end. However, some very low magnitude ( < 10 μV) and very narrow bottom ( < 0.5 second) interference wave was detected during the procedures above. No surface electromyogram signal was detected when gastrocnemius muscle was in the passive motion. These findings suggest that rabbit gastrocnemius muscle surface electromyogram signal is generated before the muscle contraction. It is a combination of drive electrical signal transmitted to muscles, which is based on upper motor center, supplemented by lower motor center. Surface electromyogram signal cannot be generated by muscle contraction itself. Related Articles | Metrics

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Puerarin combined with estradiol for treatment of osteoporosis in ovariectomized rats Li Hai, Wang Jin-hua, Huang Hai-ling, Li Biao 2011, 15 (46):  8698-8701.  doi: 10.3969/j.issn.1673-8225.2011.46.035 Abstract ( 263 )   PDF (336KB) ( 547 )   Save BACKGROUND: Estradiol produces great adverse effects in treatment of postmenopausal osteoporosis. OBJECTIVE: To investigate the effects of small dose of estradiol combined with estradiol on bone tissue of ovariectomized rats. METHODS: A total of 120 healthy female rats were randomly and evenly divided into five groups: sham-operated (only resection of a small block of para-ovarian fat pad), ovariectomized (without drugs), puerarin, estradiol, and puerarin + estradiol. One week later, rats from the puerarin, estradiol and puerarin + estradiol groups were subcutaneously injected with puerarin (50 mg/kg, once a day), estradiol (200 μg/kg, twice a week), estradiol puerarin (25 mg/kg, once a day) + (100 μg/kg, twice a week), respectively. RESULTS AND CONCLUSION: In the ovariectomized group, rat bone tissue was sparse, bone calcium and phosphorus levels as well as bone mineral density were significantly lower compared with sham-operated group (P < 0.01). After estradiol and/or puerain treatment, rat bone tissue morphology was obviously improved, and bone calcium and phosphorus levels and bone mineral density were significantly increased. The above-mentioned indices were similar after small dose of puerarin combined with estradiol treatment and high dose of puerarin combined with estradiol treatment. These findings suggest that small dose of estradiol combined with estradiol treatment yields satisfactory curative effects in treatment of osteoporosis in ovariectomized rats. Related Articles | Metrics

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Effects of the prescription for tonifying kidney and strengthening spleen on caspase-3 and caspase-8 expression in skeletal muscle of ovariectomized rats Li Ying,Bai Bo, Huang Hong-xing, Wu Huo-yan, Zhang Shu-jiang, Wang Guang-wei 2011, 15 (46):  8702-8705.  doi: 10.3969/j.issn.1673-8225.2011.46.036 Abstract ( 259 )   PDF (640KB) ( 403 )   Save BACKGROUND: Studies have shown that there is a close relationship between skeletal muscle and osteoporosis. Apoptosis is a main reason of skeletal muscle aging, which may be mediated by the proteins of caspase family. OBJECTIVE: To study the mechanism of the prescription for tonifying kidney and strengthening spleen on preventing and treating osteoporosis and observe the regulation of the prescription on caspase-3 and caspase-8 expression in skeletal muscle of ovariectomized rats. METHODS: Forty-eight female non-pregnant SD rats (6 months) were randomly and evenly divided into four groups: control, ovariectomized, estrogen and traditional Chinese herb. After establishing model, the rats of traditional Chinese herb group were intragastrically administered with the prescription for tonifying kidney and strengthening spleen (2.979 g/kg). The rats of estrogen group received an intragastric administration of estradiol valerate (0.104 mg/kg). The rats of the other groups were intragastrically given distilled water, once a day. RESULTS AND CONCLUSION: Dual energy X-ray absorptiometry results showed that compared with control group, bone mineral density (BMD) and bone mineral content (BMC) in rat left femur were significantly lower in the model group (P < 0.01). The BMD was significantly greater in the estrogen and traditional Chinese herb groups than in the model group (P < 0.05) and the BMD and BMC were similar between the estrogen and traditional Chinese herb groups (P > 0.05). Enzyme linked immunosorbent assay results showed that compared with the control group, caspase-3 and caspase-8 expression in the skeletal muscle was significantly increased in the model group (P < 0.05); caspase-3 expression was significantly decreased in the traditional Chinese herb group (P < 0.05), and caspase-8 expression was significantly decreased in the estrogen group (P < 0.05). These findings suggest that the prescription for tonifying kidney and strengthening spleen exhibits obvious therapeutic effects on ovariectomy-induced osteoporosis, can increase bone mineral density and obviously decrease caspase-3 expression, but it can not markedly decrease caspase-8 expression. This indicates that the prescription does not inhibit apoptosis by death receptor signal pathway. Related Articles | Metrics

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Effects of tetramethylpyrazine on expression of connective tissue growth factor, nuclear factor kappa B and its related gene products in tumor necrosis factor-alpha stimulated hepatic stellate cells Li Yan-fang, Li Xiao-sheng 2011, 15 (46):  8706-8711.  doi: 10.3969/j.issn.1673-8225.2011.46.037 Abstract ( 258 )   PDF (896KB) ( 359 )   Save BACKGROUND: Previous studies have shown that tetramethylpyrazine (TMP) can decrease expression of connective tissue growth factor (CTGF), nuclear factor kappa B (NF-κB) and delay the formation of hepatic fibrosis. But the mechanism and the correlation of CTGF and NF-κB was not clear.            OBJECTIVE: To investigate the effects of TMP on expression of CTGF, NF-κB and its related gene product interleukin-6 (IL-6) in tumor necrosis factor-alpha (TNF-α) stimulated hepatic stellate cells. METHODS: Hepatic stellate cell-T6 (HSC-T6) was cultured to logarithmic growth phase for experiments. Cells were divided into four groups: control group: only cells were added; TNF-α group: 10 μg/L TNF-α was added; TMP intervention group: pretreated with 10 μg/L TNF-α for 30 minutes and then 50,100, 200, 400, 600, 1 000 mg/L TMP was added; PDTC group: pretreated with   10 μg/L TNF-α for 30 minutes, and then stimulated with 18 μmol/L PDTC, a blocker of NF-κB. RESULTS AND CONCLUSIONS: MTT assay results showed that TMP inhibited the proliferation of HSC-T6 at a concentration of 100, 200, 400, 600, and 1 000 mg/L in a concentration dependent manner (P < 0.05). Immunocytochemical staining and western blot analysis showed that 200, 400, 600 mg/L TMP and 18 μmol/L PDTC could decrease the expression of TNF-α-induced CTGF, NF-κB and IL-6 (P < 0.01) in HSC-T6 cells and with increasing concentrations of TMP, the inhibitory effects were enhanced, and PDTC could inhibit their expression effectively. Correlation analysis showed that the expression of cTGF and NF-κB in HSC-T6 cells was positively correlated (r=0.980, P < 0.01).These data indicated that TMP could inhibit the expression of cTGF, NF-κB and IL-6 in hepatic stellate cells and inhibit the proliferation of hepatic stellate cells. Related Articles | Metrics

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Role of transforming growth factor beta 2 in cartilage formation Yang Yue, Xue Tong-yuan, Tian Jing 2011, 15 (46):  8714-8717.  doi: 10.3969/j.issn.1673-8225.2011.46.039 Abstract ( 282 )   PDF (651KB) ( 494 )   Save BACKGROUND: Transforming growth factor β2 is a family of polypeptide growth factors with multiple biological effects. It plays a role in the proliferation and differentiation of mesenchymal stem cells. It is one of the most important regulation factors for the mesenchymal stem cell proliferation and differentiation into cartilage cells. OBJECTIVE: To analyze the molecular structure of transforming growth factor β2 and its role in cartilage formation. METHODS: An online search of CNKI, VIP and PubMed database was performed for relevant articles on transforming growth factor β2 and cartilage formation of bone tissue engineering published between January 1995 and August 2011, using key words of " TGF-β2; cartilage formation; tissue engineering" in Chinese and English, respectively. Articles on the induction of chondrocyte differentiation, treatment progress of cartilage diseases, molecular structure and signal transduction pathway of transforming growth factor β2 were included. Duplicated studies, antiquated standpoint or review literatures were excluded. RESULTS AND CONCLUSION: A total of 302 articles were obtained in the initial search. According to inclusion criteria, 262 of them were excluded, 40 documents were finally selected. Analysis results show that transforming growth factor β2 can facilitate the synthesis of cartilage specific matrix, such as type II collagen and proteoglycan, by promoting the differentiation from mesenchymal stem cell to chondrocyte. Therefore, transforming growth factor β2 has an induction effect on cartilage. It regulates the proliferation and differentiation of chondrocyte with other growth factors, which makes it possible to permanently repair the cartilage defects in the clinic. Related Articles | Metrics

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Tissue-engineered repair of meniscus sports injury Pei Cai-li 2011, 15 (46):  8718-8721.  doi: 10.3969/j.issn.1673-8225.2011.46.040 Abstract ( 275 )   PDF (822KB) ( 469 )   Save BACKGROUND: The self-restoring ability of meniscus injury is poor. Tissue-engineered methods provide the possibility for the meniscus function recovery. OBJECTIVE: To review the treatment of meniscus injury, and to emphasize tissue-engineered repair of meniscus. METHODS: An online search of PubMed database and Wanfang database was performed using key words of “meniscus, treatment, the measure, materials” in both English and Chinese for articles on meniscus injury and its medical materials published between January 1980 to March 2011. Articles related to the treatment and materials of meniscus were retained. Articles published recently or in authoritative journals were of priority. RESULTS AND CONCLUSION: A total of 174 articles were obtained in initial search. According to the inclusion criteria, 35 articles were finally selected. Research shows that areas surrounding the cartilage have a certain self-repair ability. It has desirable effects on the later stage of conventional treatments for regularity laceration which preserves the tissue, and has very little influence on the conventional treatments for serious injury. The condition to make use of tissue-engineered methods for meniscus treatment is becoming increasingly diverse and matured. The selection and development of ideal material are important factors of restricting the rehabilitation effect. Related Articles | Metrics

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Application of tissue-engineered anterior cruciate ligament in repair of exercise injury Shi Hai-ping 2011, 15 (46):  8722-8725.  doi: 10.3969/j.issn.1673-8225.2011.46.041 Abstract ( 340 )   PDF (605KB) ( 283 )   Save BACKGROUND: Anterior cruciate ligament injury is extremely difficult for athletes to fully recover, and can shorten the exercise span. OBJECTIVE: To summarize the recent research progress on the application of tissue-engineered anterior cruciate ligament in repair of exercise injury. METHODS: An online search of VIP database and PubMed database was performed for relevant articles published between January 1994 and December 2010. A total of 28 articles on sports-related anterior cruciate ligament injury and tissue-engineered ligament were retrieved. The discussion was mainly focused on the recent development of tissue-engineered anterior cruciate ligament reconstruction, the sources of anterior cruciate ligament seed cells, the application of cytokine in reconstruction of anterior cruciate ligament and the basic research on tissue-engineered anterior cruciate ligament. RESULTS AND CONCLUSION: Autohealing ability of sports-related anterior cruciate ligament is extremely poor, so it is extremely difficult for athletes to recover. In recent years, research on tissue-engineered anterior cruciate ligament has made rapid progress with prospect and extensive application. But with regard to application, the optimal clinical outcomes of anterior cruciate ligament repair and reconstruction can only be achieved by organically blending the selection of tissue-engineered seed cells, the construction of scaffold materials with the basic research on tissue-engineered anterior cruciate ligament, as well as using growth factor properly. Related Articles | Metrics

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Related factors of cranial and maxillofacial bone defect repair Sun Yong, Liu Liu, He Xiao-guang, Wang Fu-ke, Li Yu-xiao, Zhong Ling 2011, 15 (46):  8726-8729.  doi: 10.3969/j.issn.1673-8225.2011.46.042 Abstract ( 309 )   PDF (559KB) ( 512 )   Save BACKGROUND: Repairing cranial and maxillofacial bone defect with tissue-engineered bone can solve the problem of lacking bone source and promote the culture of tissue-engineered bone and assist in repairing bone defects using gene therapy. OBJECTIVE: To summarize the related factors of cranial and maxillofacial bone defect repair and introduce the research progress in this type of factors, providing reference evidence for repairing cranial and maxillofacial bone defect using tissue-engineered METHODS: Taking “tissue engineering, bone defect, gene therapy, growth factor” as key words, a computer-based online retrieval was performed to search papers published between January 2000 and January 2011 in PubMed and CBM databases in English and Chinese. Papers regarding related factors of cranial and maxillofacial bone defect repair were included and papers with repeated contents were excluded. RESULTS AND CONCLUSION: A total of 670 papers were initially retrieved. According to inclusion and exclusion criteria, 33 papers were included in the final analysis. There are more and more causes for cranial and maxillofacial bone defects and lacking bone source is a primary problem of restricting cranial and maxillofacial bone defect repair. In vitro construction of tissue-engineered bone is a primary method for solving this problem. Cranial and maxillofacial bone formation involves selection of related factors and construction of tissue-engineered bone, and multi-gene therapy is a promising method. Related Articles | Metrics

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Mechanism of adaptation to exercise in skeletal muscle Zhang Xiao-feng, Shi Song, Wei Yong-wang 2011, 15 (46):  8730-8733.  doi: 10.3969/j.issn.1673-8225.2011.46.043 Abstract ( 407 )   PDF (617KB) ( 478 )   Save BACKGROUND: Study on the mechanism of adaptation to exercise in skeletal muscle has great significance for the improvement of sports performance, the prevention and treatment of some metabolic disorders. OBJECTIVE: To discuss the mechanism of adaptation to exercise in skeletal muscle. METHODS: An online search of PubMed database and CNKI was performed for relevant articles published before March 2011, using key words of “skeletal muscle, exercise, adaptation, cytoskeleton, dystrophin” in English and Chinese, respectively. A total of 56 articles were collected. Articles on the mechanism of adaptation to exercise in skeletal muscle were included. A number of 31 articles were retained after excluding the repetitive ones. RESULTS AND CONCLUSION: Intense exercise can cause stress reaction in muscle structure and cell metabolism, such as muscle damage and oxidative stress. High-strength eccentric activities may lead to the damage of muscle ultrastructure, but the muscle shows a remodeling response after exercise-induced damage. Exercise training can promote various adaptations of healthy individuals to nitric oxide system. These adaptations can somehow enhance the biological availability of skeletal muscle, consequently increase the exercise capacity and provide protection to the cardiovascular system. At present, the mechanism of adaptation to exercise in human skeletal muscle is still unclear. Related Articles | Metrics

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Changes in morphological characteristics of chin soft tissue profile before and after class Ⅲ malocclusion treatment Wang Ling, Li Zhen-ya 2011, 15 (46):  8734-8736.  doi: 10.3969/j.issn.1673-8225.2011.46.044 Abstract ( 261 )   PDF (557KB) ( 591 )   Save BACKGROUND: Previous studies regarding morphological characteristic of mental region are performed to quantitatively analyze the soft and hard tissue of tooth and mental region before and after treatment. OBJECTIVE: To investigate the changes in morphological characteristic of chin soft tissue profile before and after class  Ⅲmalocclusion treatment. METHODS: The sample was composed of 48 adolescents aged 12-16 years who presented class Ⅲ malocclusion. Cephalograms were analyzed for all subjects, and the morphology of the symphysis before and after treatment was measured. Paired student’s t test was performed through the use of SPSS16.0 software package. RESULTS AND CONCLUSION: After treatment, the length of chin (LL-Pos) decreased (P=0.000), Bs-LLPos was larger (P=0.000), ∠LL-Bs-Pos (P=0.000) and ∠Bs-Pos-Mes (P=0.004) were decreased compared with before treatment. After treatment, all cephalogram dates of chin convexity were smaller compared with before treatment (P < 0.05). The thickness of chin soft tissue was decreased after treatment compared with that before treatment (P < 0.05). The morphological characteristic of chin soft tissue became better after treatment compared with before treatment. Related Articles | Metrics

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Association between A163G polymorphism of the osteoprotegerin gene and bone mineral density in the Chinese elderly from Beijing Area Wei Ya-nan, Liu Jie, Chen Ling-xia, Jia Rong, Miao Yi-de 2011, 15 (46):  8737-8740.  doi: 10.3969/j.issn.1673-8225.2011.46.045 Abstract ( 241 )   PDF (640KB) ( 321 )   Save BACKGROUND: Osteoprotegerin gene is the candidate gene of osteoporosis, and the ethnic divergence of gene polymorphisms exists.  OBJECTIVE: To investigate the association between A163G polymorphism of the osteoprotegerin gene and bone mineral density in the elderly in Beijing. METHODS: A total of 307 elderly were included in this study. The bone mineral density in proximal femur, lumbar vertebra and forearm was determined by dual energy X-ray absorptiometry and DNA was extracted from peripheral blood. The A163G polymorphism of the osteoprotegerin gene was assessed by polymerase chain reaction-restriction fragment length polymorphism analysis and gene sequencing was performed. Differences in bone mineral density among different genotypes were compared using one-way analysis of variance. Bone mineral density-related factors were analyzed using multiple stepwise linear regression analysis.  RESULTS AND CONCLUSION: In elderly women, the AA genotype of the A163G was associated with significantly lower bone mineral density at lumbar and femoral inter area; while in elderly men, no association was found between the A163G polymorphisms and bone mineral density. These findings suggest that A163G genotypes in the osteoprotegerin gene promoter are associated with bone mineral density in elderly women. Related Articles | Metrics