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11 June 2010, Volume 14 Issue 24 Previous Issue    Next Issue

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In vitro differentiation from bone marrow mesenchymal stem cells into chondrocytes induced by transforming growth factor-beta combined with bone morphogenetic protein-2 Zhang Qing-lin, Lü Hui-cheng, Wu Yi-min 2010, 14 (24):  4371-4375.  doi: 10.3969/j.issn.1673-8225.2010.24.001 Abstract ( 85 )   PDF (406KB) ( 451 )   Save BACKGROUND: Multiple studies have reported that transforming growth factor-β1 (TGF-β1) induced differentiation from bone marrow mesenchymal stem cells (BMSCs) into chondrocytes; however, reports addressing bone morphogenetic protein-2 (BMP-2) inducing the differentiation remain less. OBJECTIVE: To verify the possibility and interaction of differentiation from BMSCs into chondrocytes induced by TGF-β1 and BMP-2. METHODS: BMSCs were derived from healthy Wistar rats using adherent methods. The BMSCs were assigned into four groups: TGF-β1+BMP-2, TGF-β1, BMP-2, and control groups. At 14 days after induction, alcian blue staining and dimethyl-methylene blue chromatometry were employed to determine the glycosaminoglycan (GAG) level, and immuno-histochemistry was used to detect the expression of specific type II collagen. RESULTS AND CONCLUSION: In the three experimental groups, alcian blue staining and type II collagen immunohistochemistry examinations showed that chondyocytes could express glycosaminoglycan (GAG) and type II collagen, and the GAG detection showed that the expression of GAG gene was significantly greater in the TGF-β1+BMP-2 group than in the TGF-β1 group and BMP-2 group (P < 0.05). TGF-β1 combined with BMP-2 was more effective for promoting the differentiation from BMSCs into chondrocytes, which might be used as the better source of the seed cells of cartilage tissue engineering. Related Articles | Metrics

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Biological character analysis of nucleus pulposus cells of the human normal intervertebral disc at different passages in vitro: Choice of seed cells for tissue engineered intervertebral disc Ye Dong-ping, Liang Wei-guo, Dai Li-bing, Shen Yan 2010, 14 (24):  4376-4379.  doi: 10.3969/j.issn.1673-8225.2010.24.002 Abstract ( 126 )   PDF (346KB) ( 311 )   Save BACKGROUND: Seed cells used for studying tissue engineered intervertebral disk are mainly derived from in vitro cultured nucleus pulposus. The in vitro cultured nucleus pulposus will affect aging at a certain passage. The aging cells lost growth ability, and were not suitable for the study of tissue engineering. OBJECTIVE: To compare the different passages of the normal nucleus pulposus cells of the morphology, proliferation characteristics and major extracellular matrix gene expression. METHODS: Human normal intervertebral disc nucleus pulposus cells were isolated and cultured using enzyme digestion method. Light microscopy, electron microscopy and other morphological methods were used to observe the general morphology and ultrastructure of the cells. XTT experiment was used to measure the differences in growth kinetics and expression of type Ⅱ collagen and glycosaminoglycan mRNA among different passages of cells. RESULTS AND CONCLUSION: Before the third passage, cells were rich in cytoplasm, presented polygonal, short spindle-shaped apperance, rich organelles in the cells, with normal morphology, less mitochondrial content and more ribosome content, which showed that the cells metabolism were exuberant vitality. Growth curve and the experimental XTT absorbance also showed that the cell proliferation was exuberant vitality, with high extracellular matrix secretion. The fourth passage of cells evolved to the long-spindle, and gradually showed fibroblast morphology, indicating cells started to present “throwback” phenomenon, also known as aging. Cell growth slowed down, the sixth and seventh passages appeared as spindle morphology, with the presence of rough endoplasmic reticulum, more mitochondria, growth stagnation, decreased cell viability and decreased extracellular matrix significantly. Results indicated that in vitro culture conditions, human nucleus pulposus cells before the third passage showed good cell morphology, proliferation ability, suitable as seed cells for tissue engineering of intervertebral disc, while the cell proliferation after fourth passage was gradually weakened and cannot be used as seed cells for tissue engineering of intervertebral disc. Related Articles | Metrics

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Effects of combining estrogen and progesterone on bone mineral density and histomorphometry in ovariectomized rats Liang Min, Zhang Jie, Liang Xing-huan, Luo Zuo-jie 2010, 14 (24):  4380-4384.  doi: 10.3969/j.issn.1673-8225.2010.24.003 Abstract ( 120 )   PDF (272KB) ( 511 )   Save BACKGROUND: Treatment of menopausal osteoporosis using estrogen combined with progesterone not only build up the therapeutic effect, but also reduce the side effect of estrogen, thus attracting more and more attention. OBJECTIVE: To observe the combining effects of 17β-estradiol and levonorgestrel on bone mineral density (BMD) and histomorphometry in ovariectomized SD rats. METHODS: Female SD rats were randomly assigned to the sham-operated group (sham group), ovariectomized group (OVX group), 17β-estradiol group (EST group), levonorgestrel group (LNG group) and 17β-estradiol combined with levonorgestrel group (EST+LNG group). After ablating bilateral ovarian model, EST, and EST+LNG group rats were administrated 17β-estradiol (10 μg/kg per day), LNG group and EST+LNG group rats were administrated levonorgestrel(60 μg/kg per day). The rats were killed 12 weeks later. The BMD of whole body was measured by bone densitometry. The fourth lumbar vertebra sections were cut for bone histomorphometry analysis. The biochemical bone markers were measured. RESULTS AND CONCLUSION: BMD and weight of thighbone in OVX group and LNG group were lower than of the sham, EST and EST+LNG groups (P < 0.05). But BMD and weight of thighbone had no significant difference between the EST group and EST+LNG group (P > 0.05). Compared with the sham group, the bone trabecular area percentage, thickness and number in OVX group and LNG group were decreased (P < 0.05), while the bone trabecular resolution, bone mineral apposition rate, bone surface area formation rate, bone volume formation rate and bone mineral apposition rate in OVX group and LNG group were increased (P < 0.05). Compared with OVX group, the bone trabecular area percentage, thickness and number in EST group and EST+LNG group were increased (P < 0.05), while the bone trabecular resolution, bone surface area formation rate, bone volume formation rate in EST group and EST+LNG group were decreased (P < 0.05). The differences between the EST group and EST+LNG group had no significant differences (P > 0.05). All results demonstrated that combining 17β-estradiol with levonorgestrel can improve BMD and reverse the high bone turnover rate, which have no harm to bone formation. Related Articles | Metrics

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Different staining methods for rat knee articular cartilage Gao Gai-xia, Wei Xiao-chun, Li Kai, Wei Jian-ping 2010, 14 (24):  4385-4389.  doi: 10.3969/j.issn.1673-8225.2010.24.004 Abstract ( 431 )   PDF (346KB) ( 2284 )   Save BACKGROUND: Extensive research has been made concerning articular cartilage by domestic and foreign scholars. Different staining methods are needed to analyze the results of the research, but there was less research applying various staining methods to cartilage research or exploring staining mechanism. OBJECTIVE: To explore the advantages and disadvantages of rat knee joint with different staining methods. METHODS: Articular cartilage of the normal rat knee joint was sampled. The structure of normal cartilage was observed through the histological staining of hematoxylin-eosin, safranin O, alcian blue, toluidine blue, safranin-alcian blue, as well as safranin-fast green. RESULTS AND CONCLUSION: Tidemarks with blue, red and blue colors could be observed in hematoxylin-eosin, safranin O and toluidine blue staining, respectively. The stain intensity of Safranin O and toluidine blue staining were increased toward tidemark. Thus, Safranin O staining was superior to other methods in observing tidemark. The four-layer structures of cartilage were exhibited clearly, with arranged chondrocytes columnar and basophilic matrix in hematoxylin-eosin staining. The safranin O staining showed that the four-layer structure of cartilage was clear, and the matrix at bottom was stained redder than other places. Alcian blue staining showed that, peripheral chondrocytes were strong stained at pH1.0, which stained deeper at PH2.5. The cartilage structure was not clearly in toluidine blue staining, with clearly nuclei and almost no coloring cytoplasm, but matrix was stained light blue purple. Safranirn-alcian blue staining showed that the cartilage surface and matrix exhibited different red, which was deep red at the bottom zones, but the surrounding of chondrocytes was stained blue. There was obviously contrast between cartilage tissue and bone tissue in safranin O staining, which showed red in the former and green in the latter. The results demonstrated that the 4-layer structure of cartilage can be observed by all above methods, especially safranin O staining, which is excellent in observing cartilage layers and tidemark structures. Hematoxylin-eosin staining provides better outcomes than other methods in observing morphologic changes of chondrocyte. Related Articles | Metrics

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Effect of estrogen on vascular endothelial growth factor expression in rat osteoblasts Pan Jin-ping, Yin Yun-sheng, Li Peng-biao, Wen Wei-jing 2010, 14 (24):  4390-4392.  doi: 10.3969/j.issn.1673-8225.2010.24.005 Abstract ( 143 )   PDF (290KB) ( 334 )   Save BACKGROUND: Vascular endothelial growth factor (VEGF) is a major angiogenesis factor, characterizing by promoting cell division, proliferation, cytoplasm calcification and aggregation, as well as inducing angiogenesis; on the other hand, it also adjusts proliferation, differentiation, and functional activity of osteoblast and osteoclast. OBJECTIVE: To investigate the effect of estrogen on VEGF expression in rat osteoblasts. METHODS: Calvaria osteoblasts derived from 48-hour-old rats were cultured to obtain osteoblast. RT-PCR was employed to detect the VEGF mRNA expression following induction of 17β-estradiol at concentrations of 10-10, 10-8, 10-6, and 10-4 mol/L. RESULTS AND CONCLUSION: RT-PCR demonstrated that VEGF mRNA was stably expressed in primary cultured osteoblast. Semi-quantitative analysis demonstrated that VEGF mRNA expression was in a certain law following 17β-estradiol induction at varying concentrations, i.e., with the increasing concentration of 17β-estradiol, VEGF mRNA expression was increased. At 10-6 mol/L, the VEGF mRNA expression peaked, and when 17β-estradiol was over this dose, the expression was gradually decreased. This suggested that estrogen promoted VEGF mRNA expression, showing a dose-dependent manner. Related Articles | Metrics

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Establishment and evaluation of a modified animal model of steroid-induced avascular necrosis of femoral head Li Chuan-jiang, Wang Wan-ming, Zhuang Yan-feng, Li Zheng, Li Jie 2010, 14 (24):  4393-4397.  doi: 10.3969/j.issn.1673-8225.2010.24.006 Abstract ( 95 )   PDF (500KB) ( 391 )   Save BACKGROUND: So far, the pathogenesis of steroid-induced avascular necrosis of femoral head was still unclear; furthermore, the valid animal model was lack, and the developments of different treatments were based on the animal models, which could simulate the natural pathogenesis course. OBJECTIVE: To explore the feasibility of setting up animal models of steroid-induced avascular necrosis of femoral head by combining low-dosage methylprednisolone (MPS) with equine serum (ES). METHODS: Twenty healthy Japanese white rabbits were randomly divided into experimental group and control group. Experimental group was intravenously injected with ES, and another injection of ES was performed two weeks later. MPS was intramuscularly injected at 24 hours after the first ES injection, once a week for 8 weeks in total. Control group was injected with the same dose of saline. Hematologic studies were performed before the first ES injection, 2, 4, and 8 weeks after MPS injection. Meanwhile, other examinations were carried out including magnetic resonance imaging (MRI), hematoxylin-eosin staining, and transmission electron microscope at 4 and 8 weeks after MPS injection, respectively. RESULTS AND CONCLUSION: Compared with control group, triacylglycerol and cholesterol levels were significantly increased in the experimental group after 2, 4, and 8 weeks (P < 0.05), but partial thromboplastin time was significantly decreased (P < 0.05). MRI in the experimental group after 4 weeks demonstrated that local signals were changed in some animals and subcortical hemorrhage was observed after 8 weeks. Compared with control group, necrotic bone marrow cell debris was located surrounding bone marrow in the experimental group after 4 and 8 weeks, and blank bone lacuna rate was gradually increased (P < 0.01). Structure of some osteocyte was unclear in the experimental group after 4 weeks, and large vacuoles were observed. After 8 weeks, part of osteocyte had caryorrhexis and caryolysis, apoptotic cells appeared abundantly, and the structure of bone collagen was confused. Combination of ES and steroid significantly increased incidence of osteonecrosis in rabbits and contributed to set up animal models of steroid-induced avascular necrosis of femoral head. Related Articles | Metrics

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Primary culture of fibroblasts from cochlear duct membrane of neonatal C57 mouse using enzyme digestion and differential adhesion methods   Wang Ting-kuo, Sun Hong 2010, 14 (24):  4398-4401.  doi: 10.3969/j.issn.1673-8225.2010.24.007 Abstract ( 121 )   PDF (388KB) ( 437 )   Save BACKGROUND: There has been no specific method for culturing fibroblasts from cochlear duct membranes so far. OBJECTIVE: To provide good cell model in vitro via culturing fibroblasts from lateral wall of cochlear duct membrane of neonatal C57 mouse by enzyme digestion and differential adhesion methods and to identify its biological features. METHODS: Tissues from lateral wall of cochlear duct membrane were acquired from neonatal C57 mouse using operating microscope. Fibroblasts from lateral wall of cochlear duct membrane were cultured by trypsin digestion combined with differential adhesion methods. The growth condition was observed by inverted phase contrast microscope and hematoxylin-eosin staining, and cell growth curve was drawn, then the immunocytochemistry was employed to classify cell types. RESULTS AND CONCLUSION: After passage and purification, the shape of fibroblasts from lateral wall of cochlear duct membrane was fusiform and triangle, with “S” type cell growth curve. Immunocytochemiscal detection showed that vimentin could be detected in cultured cells, and brown cytoplasmic pigment could be seen. The results demonstrated that fibroblasts can be cultured form lateral wall of cochlear duct membrane of mice.  Related Articles | Metrics

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Construction and identification of recombinant adenovirus vector co-expressing human bone morphogenetic protein 2 and fibroblast growth factor 2 genes    Guo Wei-tao, Liu Si-jing, Wang Hui, Xiao Qi-xian, Chen Zi-qiu, Huang Yun, Wang Bin 2010, 14 (24):  4402-4406.  doi: 10.3969/j.issn.1673-8225.2010.24.008 Abstract ( 115 )   PDF (388KB) ( 336 )   Save BACKGROUND: The upstream and downstream genes can be co-transcribed by internal ribosome entry site (IRES) sequence, by which human bone morphogenetic protein 2 (hBMP2) and human fibroblast growth factor 2 (hFGF2) genes can be highly co-expressed, thus, it provides a new way for repairing bone defects. OBJECTIVE: To construct the recombinant adenovirus vector co-expressing hBMP2 and hFGF2 depending on IRES sequence by λ phage-site specific recombination systems and to verify the correct recombination. METHODS: BMP-2cDNA and FGF-2 cDNA were amplified by PCR, and were subcloned into the pIRES2-EGFP plasmid to obtain phBMP2-IRES-hFGF2 plasmid. It was subcloned into entry vector pDONR221 by BP reaction and subcloned into pAd/CMV/V5-DEST vector by LR reaction and linearization transfection 293 cells to obtain pAd-hBMP2-IRES-hFGF2. RESULTS AND CONCLUSION: The target gene of hBMP2-IRES-hFGF2 was transferred into AD/CMV/V5-DEST vector correctly and it was confirmed by sequencing. The expression clone pAd-hBMP2-IRES-hFGF2 was packaged into maturated adenovirus successfully; the titer of Ad-hBMP2-IRES-hFGF2 was 2.82×1010 ifu/mL. The results demonstrated that adenovirus vector co-expressing hBMP2 and hFGF2 are constructed successfully.  Related Articles | Metrics

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Segmental artery ligation for establishment of local spinal cord ischemic injury model in rabbits Tan Mei-yun, Guo Xing, Wang Yuan-hui 2010, 14 (24):  4407-4410.  doi: 10.3969/j.issn.1673-8225.2010.24.009 Abstract ( 203 )   PDF (371KB) ( 424 )   Save BACKGROUND: Previous studies showed that a stable model of spinal cord injury can be established using a method of lumbar arteries ligation in retroperitoneum. However, the precise ligation area is unclear.  OBJECTIVE: To establish a rabbit model of local spinal cord ischemic injury produced by segmental artery ligation. METHODS: A total of 16 adult rabbits were randomly divided into the control and model groups, with 8 animals in each group. Rabbits in the model group experienced ligation of segment artery in retroperitoneum. There was no treatment in the control group. The nerve function of spinal cord was evaluated by modified Tarlov score and the pathological changes of lumbar segmemal spinal cord were observed with the hematoxylin-eosin staining at 3 days after operation. RESULTS AND CONCLUSIONS: Paralysis was evidenced in the model group and the Tarlov scores were smaller than that of the control group (P < 0.05). Acute inflammatory lesion and neuronal necrosis were observed in the model group. Results showed that a stable model of local spinal cord ischemic injury can be established via ligating L1-3 lumbar arteries approach retroperitoneum. Related Articles | Metrics

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A rat model of liver transplantation constructed by cryopreservation    Yu Qiang, Shi Xian-jie, Li Hai-lin, Dong Jia-hong 2010, 14 (24):  4411-4415.  doi: 10.3969/j.issn.1673-8225.2010.24.010 Abstract ( 113 )   PDF (402KB) ( 345 )   Save BACKGROUND: Liver transplantation is the most effective treating method for end-stage liver disease. Biliary lesion following transplantation is one of primary problems for liver transplantation. OBJECTIVE: To develop a model of liver transplantation in rats to study the effect of cryopreservation time on bile ducts following liver transplantation. METHODS: A total of 120 healthy, male, SD rats. According to the random pair principle, rats with smaller weight served as donors. The donor livers were preserved at UW solution at 4 ℃ for 2, 8, 16 hours and transplanted to the recipients. Based on the classic “two cuff” technique for orthotropic liver transplantation, end-to-end anastomosis between the common hepatic artery of donor and recipient by the modified “Gao stent” was performed to reconstruct hepatic artery blood flow. The operative duration and successful transplantation ratio were recorded. The levels of serum alamine aminotransferase, total bilirubin, and alkaline phosphatase were determined at 3 and 7 days after transplantation. At the same time, the histopathological changes were observed.  RESULTS AND CONCLUSION: In total 55 rats were performed orthotopic liver transplantation, and the successful transplantation ratio was 93%. The success ratio of 2-, 8- and 16-hour group was 100% (9/9), 83% (10/12) and 73% (8/11), respectively. Prolonged cryopreservation time can cause the severe bile duct injury. Bile duct histopathology score showed differences between groups were significant (P < 0.05). The model in rats with grafts undergoing 16-hour cryopreservation provides suitable model for studying the effects of cryopreservation on bile ducts after liver transplantation. Related Articles | Metrics

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Establishment of two-dimensional electrophoresis separation system for proteomes of human kidney tubular epithelial cells   Zhang Chun-yan, Liu You-ping, Yang Ye, Gong Shu, Li Hong 2010, 14 (24):  4416-4420.  doi: 10.3969/j.issn.1673-8225.2010.24.011 Abstract ( 166 )   PDF (540KB) ( 333 )   Save BACKGROUND: Two-dimensional electrophoresis (2-DE) separation technology is one of core technologies for proteomics studies. However, the resolution of 2-DE map is highly altered due to different experimental conditions. Therefore, a 2-DE map with high resolution can be obtained via the optimization of key experimental conditions for a definite proteome sample. OBJECTIVE: To establish an effective separation system of 2-DE for the proteome of HK-2 human renal tubular epithelial cells. METHODS: HK-2 human renal tubular epithelial cells were cultured routinely. The proteins were extracted from HK-2 cells after lysing, and separated by 2-DE on the standard experimental condition with an optimization for each important experimental parameter. Electrophoresis parameters recommended by Bio-Rad Corporation were adjusted adequately. In the process of silver staining, a sufficient washing operation ensured a weak background of 2-DE maps. The gray densities and numbers of protein spots, resolution and contaminating strands in 2-DE maps were determined. RESULTS AND CONCLUSION: A 2-DE separation system for proteome of human renal tubular epithelial cells was successfully established. The optimized experimental conditions were selected finally as follows: The lysis buffer containing 1% TBP, 4% CHAPS, 0.2% Bio-Lyte, 40 mmol/L Tris, 8 mol/L urea, 2 mol/L thiourea was applied to extract proteins from cells, and the passive rehydration proposal of samples was used to help the separation of large molecular weight proteins on the pH 4-7 IPG gels. Electrophoresis parameters recommended by Bio-Rad Corporation were adjusted adequately. In the process of silver staining, a sufficient washing operation ensured a weak background of 2-DE maps. Under the optimized conditions, 2-DE maps with high resolution have been reproducibly obtained. Related Articles | Metrics

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Construction and prokaryotic expression of canis melanocortin receptor-3 Xie Jian-zhong, Wang Guang-chuan, Ba Cai-feng 2010, 14 (24):  4421-4424.  doi: 10.3969/j.issn.1673-8225.2010.24.012 Abstract ( 175 )   PDF (375KB) ( 296 )   Save BACKGROUND: Effects of melanocortin receptor (MCR) on in vivo energy balance arose more and more attention. MC3R gene mutation plays an important role in weight gaining, but there are few studies concerning MC3R, and no monoclonal antibody has been reported to detect MC3R expression at protein level.  OBJECTIVE: To construct an expression vector pGEX-4T-1-cMC3R and to induce its expression in E. coli. METHODS: Using the canis MC3R DNA as template, specific primers were designed. After the PCR amplification, the product was cloned into pMD18-T vector, and identified by double enzyme digestion. The target gene was subcloned into pGEX-4T-1 vector. The recombinant pGEX-4T-1-cMC3R was transferred to E. coli DH5α, and then it was identified with double restriction enzymes digestion analysis and DNA sequencing. The recombinant pGEX-4T-1-cMC3R which was identical to what released in GeneBank was transformed into E. coli BL21. The expression of canis MC3R protein was analyzed by Western blot after IPTG induction. RESULTS AND CONCLUSION: The prokaryotic plasmid pGEX-4T-1-cMC3R was constructed successfully. Canis MC3R sequence was mainly identical to what released in GeneBank. After induction, the recombinant pGEX-4T-1-cMC3R could express canis MC3R fusion protein in E. coli BL21. This experiment not only provides material basis for construction of canis MC3R monoclonal antibody, but also benefits the structures and physiologic functions studies of canis MC3R protein. Related Articles | Metrics

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Constructing a recombinant adenovirus vector pAd-LMP-1 using Gateway technology  Gu Hong-sheng, Cheng Zhi-an, Li Zhen-yu, Zhou Wen-yu, Zou Xiao-ying 2010, 14 (24):  4425-4429.  doi: 10.3969/j.issn.1673-8225.2010.24.013 Abstract ( 233 )   PDF (389KB) ( 486 )   Save BACKGROUND: The construction of adenovirus vector using Adeasy system is tedious with poor results due to the multiple restriction enzyme digestion, linkage, and screening. OBJECTIVE: To construct a recombinant adenovirus vector containing LMP-1 gene using Gateway technology. METHODS: Total RNA was extracted from rat osteoblasts and the LMP-1 gene was acquired by RT-PCR, the LMP-1 gene and entry vector pENTR/D-TOPO were used to create the entry clone with the directional TOPO clone technology, then the entry clone and the expression vector were used to create the expression clone through the LR recombination reaction; at last the adenovirus expression clone was linearized by PacI and transfected to the 293A cell line. The target gene was compared to LMP-1 in the Genebank (Gene number: AF095585), and the expression of pAd-LMP-1 was observed. RESULTS AND CONCLUSION: The electrophoresis showed that the extracted RNA was integrated, and the purity was accorded with the requirement. LMP-1 gene was successfully acquired. The entry clone was verified by enzymes digestion, and sequencing. The constructed pAd-LMP-1 recombination adenovirus vector was successfully packaged in 293A cells. This result demonstrated that it is easy and rapid to obtain pAd-LMP-1 by constructing pAd-LMP-1 recombination adenovirus vector using the Gateway technology. Related Articles | Metrics

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Comparison of four methods for DNA extraction from paraffin-embedded tissues Yuan Ya-ting, Jiang Yue-xin, Yin Xiao-wen, Jiang Xing-tang 2010, 14 (24):  4430-4434.  doi: 10.3969/j.issn.1673-8225.2010.24.014 Abstract ( 149 )   PDF (350KB) ( 780 )   Save BACKGROUND: Because of the damage of formalin-fixed paraffin-embedded tissue (FFPET) to DNA during the production and preservation, it is difficult to extract high quality and sufficient DNA for the following researches such as polymerase chain reaction (PCR). Therefore, it is essential and urgent to investigate an effective and economic method for extraction of DNA from FFPET. OBJECTIVE: To investigate the influence of four DNA extraction methods for FFPET on DNA quality, and to identify the optimal method for DNA extraction from FFPET. METHODS: Twenty formalin-fixed, paraffin-embedded non-small cell lung cancer (NSCLC) tissue specimens were selected and divided into four groups according to the method for DNA extraction. Four methods included xylene dewaxing and phenol-chloroform method, improved TES dewaxing with water-bath and phenol-chloroform method, genomic DNA extraction kit, improved TES dewaxing with water-bath and genomic DNA extraction kit. The quality of obtained DNA was analyzed by electrophoresis and PCR amplification. RESULTS AND CONCLUSION: For paraffin-embedded tissue specimens, better DNA fragments could be obtained by using the improved TES dewaxing with water-bath and phenol-chloroform method and improved TES dewaxing with water-bath and genomic DNA extraction kit. The A260/A280 ratio of DNA obtained by using improved TES dewaxing with water-bath and phenol-chloroform method and improved TES dewaxing with water-bath and genomic DNA extraction kit were statistically different from genomic DNA extraction kit (P < 0.01). There was no significant difference in efficiency among four mehtods (P > 0.1). The aim trips were as bright as the positive control when using the DNA extraction by improved TES dewaxing with water-bath and phenol-chloroform method as the template. The results demonstrated that improved TES dewaxing with water-bath and phenol-chloroform method is an optimal method for extraction of DNA from FFPET. Related Articles | Metrics

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Expression of ubiquitin-protein ligase Murf1 and nuclear factor kappa B in denervated amyotrophy intervened by passive movement Han Li-jun, Liang Bing-sheng, Wang Le, Zhang Lei 2010, 14 (24):  4435-4438.  doi: 10.3969/j.issn.1673-8225.2010.24.015 Abstract ( 142 )   PDF (416KB) ( 482 )   Save BACKGROUND: Protein degradation is a key link in delaying amyotrophy. The expressions of muscle ring finger protein (Murf1) and nuclear factor κB (NF-κB) are increased in amyotrophy, and studies have been demonstrated that passive movement can suppress the occurrence of amyotrophy. OBJECTIVE: To investigate expression pattern of Murf1 and NF-κB during denervated amyotrophy. at different time points, and to explore effects of passive movement on the expression of Murf1 and NF-κB in denervated amyotrophy. METHODS: Sciatic nerves of rats were cut down in the denervated and passive movement groups. Sham operation was performed in the sham-operated group. Rats in the passive movement group were performed passive movement (45° traction and full range flexion) at 1 day after operation, twice per day, 300 frequency per time. The expressions of Murf1 and NF-κB were determined by RT-PCR and Western blot at 2, 14 and 28 days after intervention. RESULTS AND CONCLUSION: Compared with the sham-operated group, the expressions of Murf1, NF-κB mRNA and protein in the denervated group were obviously increased at each time points (P < 0.05). Compared with the denervation group, the expressions of Murf1, NF-κB mRNA in the passive movement group were dramatically decreased (P < 0.01). The muscle wet weight was obviously decreased after denervation, which was greater in the positive movement group than that of the denervation group at 14 days (P < 0.05). The expression of mRNA and protein of Murf1 and NF-κB were negatively correlated to muscle wet weight in the denervated group (r = -0.795, P < 0.01; r = -0.834, P < 0.01). Passive movement can prevent amyotrophy by reducing the expressions of Murf1 and NF-κB. Related Articles | Metrics

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Gait and movement coordination in patients with pregnancy-related pelvic girdle pain Wu Wen-hua,Lin Xiao-cong 2010, 14 (24):  4439-4444.  doi: 10.3969/j.issn.1673-8225.2010.24.016 Abstract ( 159 )   PDF (742KB) ( 316 )   Save BACKGROUND: Walking is often impaired in pregnancy-related pelvic girdle pain (PPP), but the precise pathogenesis remains poorly understood. OBJECTIVE: To better understand the walking velocity, gait and changes in movement coordination in patients with PPP and to explore its underlying mechanism of movement pathology. METHODS: The gait kinematics of 12 healthy pregnant women and 12 pregnant women with PPP were compared, focusing on the amplitudes of transverse pelvic, lumbar, and thoracic segmental rotations, the timing and relative phase of these rotations, and the amplitude of spinal rotations. RESULTS AND CONCLUSION: The walking velocity of pregnant women with PPP was lower than that of the controls, and negatively correlated with fear of movement. While patients’ amplitudes were larger in transverse pelvic, lumbar, and thoracic segmental rotations, and there were large inter-individual differences. The spinal rotations did not differ between groups. In the patients, peak thorax rotation occurred earlier in the stride cycle at higher velocities, and relative phase was lower, probably to avoid excessive rotational torque in the sacroiliac joints and the spine. Related Articles | Metrics

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Tensile mechanical properties of middle cerebral artery in an atherosclerosis model Yu Bo, Sun Chang-jiang, Quan Tie-gang, Xu Dong-hui, Ma Hong-shun 2010, 14 (24):  4445-4448.  doi: 10.3969/j.issn.1673-8225.2010.24.017 Abstract ( 159 )   PDF (275KB) ( 607 )   Save BACKGROUND: It is necessary to understand the mechanical properties of middle cerebral artery of normal or atherosclerotic rats in order to prevent and cure atherosclerosis and cerebral hemorrhage. However, most previous studies focus on mechanical properties of normal human corpses and animals. OBJECTIVE: To compare the tensile mechanical properties of middle cerebral artery between normal and atherosclerotic rats. METHODS: Sprague Dawley rats were randomly divided into the control and model groups. The atherosclerotic models were prepared in the model group. Then rats were performed 5 mm/min tensile test at electron Shimadzu universal testing machine. The experimental data, such as maximum load, maximum displacement, maximum stress, maximum strain, as well as the stress-strain relationship were observed.  RESULTS AND CONCLUSION: Compared with the control group, the maximum load, maximum displacement, maximum stress, and maximum strain of the model group were obviously decreased (P < 0.05). The stress-strain curve changed in an exponential relationship. Accordingly, the middle cerebral artery of atherosclerotic rats exhibits distinct tensile mechanical properties from normal rats, which can not extent greatly.  Related Articles | Metrics

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Changes in contractile function of cardiocytes following overexpressing beta 2 adrenergic receptor in canines and the underlying mechanisms Shan Yu, Gong Hai-bin, Wang Lei, Wang Xiu-li, Lü Qian, Zhang Qing-lin, Liu Ying, Wang Jie, Pang Hui, Su Jing-li, Wang Zhen-quan 2010, 14 (24):  4449-4453.  doi: 10.3969/j.issn.1673-8225.2010.24.018 Abstract ( 87 )   PDF (394KB) ( 431 )   Save BACKGROUND: Previous studies have found that cardiocytes overexpressing β-adrenergic receptors (β-AR) can improve its contractile function in transgenic mice, rats and other small animals, but experimental reports about the large mammals are few. OBJECTIVE: To observe changes and mechanisms of contractile function following cardiocytes overexpressing β2-AR in canines. METHODS: Canine cardiocytes were isolated by collagenase II and transfected by adenovirus β2AR. The expression of β2AR protein of cardiocytes, the level of intracellular cyclic adenosine monophosphate (cAMP) at basic state and maximum systolic states, and the changes of contractile function were observed at 48 hours after operation.  RESULTS AND CONCLUSION: Compared with the control group, the expression of β2AR protein of cardiocytes and the level of the intracellular cAMP in the adenovirus β2AR group were increased (P < 0.05). The basal contraction amplitude of cadiocytes in adenovirus β2-AR group was also increased significantly (P < 0.05), but the max contraction amplitude of cardiocytes was not changed significantly between two groups. The study suggested that the over-expression of β2AR could increase the level of intracellular cAMP and improved the contractile function of cadiocytes in canines. Related Articles | Metrics

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Effect of Diechuang San on the experimental fracture healing in rabbits Liu Guo-hua, Wang Zhi-yi, Liu Dan-ping 2010, 14 (24):  4454-4457.  doi: 10.3969/j.issn.1673-8225.2010.24.019 Abstract ( 110 )   PDF (353KB) ( 551 )   Save BACKGROUND: The Chinese medicine Diechuang San (DCS) has the property of activating blood circulation and removing stasis, as well as deswelling and alleviating pain, which obtain good outcomes in treating soft tissue injury. Bone fracture and soft tissue injury belong to “tendon trauma” in traditional Chinese medicine. OBJECTIVE: To study the effect of DCS on experimental fracture healing. METHODS: A total of 36 male New Zealand white rabbits were prepared for 3-mm fracture bone defects models at the middle of both radiuses. Then those rabbits were separated into model control group and DCS treatment group, which were treated by bandaging and external application of DCS, once two days. Afterwards, the changes of alkaline phosphatase (ALP), Ca and P concentrations, histopathology, as well as the X-ray films were observed at 2, 4 and 6 weeks after model preparation. RESULTS AND CONCLUSION: Compared with the model group, the bone defects in the DCS group were filled quickly and the fracture healed earlier. The concentrations of serum ALP, Ca and P were greater in the first 4 weeks (P < 0.05, P < 0.01), which could promote the deposition of calcium in the fracture fragments, and promoted the fracture healing. The stimulated blood vessel was greater than the model group with larger inside diameter, and the ossification centers appeared earlier. Therefore, we presume that DCS may influence the serum ALP, Ca, P concentration changes and promote blood circulation to improve the fracture region such as the promotion of callus growth, and thus achieve the purpose of promoting fracture healing. Related Articles | Metrics

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Differences of surface morphology, element content and micro-hardness between dentinogenesis imperfecta and normal permanent dentin Xi Ge, Zhang Dong, Meng Yong-jie, Wang Jian-rong, Liu Bin 2010, 14 (24):  4458-4461.  doi: 10.3969/j.issn.1673-8225.2010.24.020 Abstract ( 99 )   PDF (438KB) ( 385 )   Save BACKGROUND: At present, the study of dentinogenesis imperfecta focuses on familial and phenotypic analysis, but the reports about physical properties of dentinogenesis imperfecta dentin are few. OBJECTIVE: To comparative the differences of surface morphology, element content and micro-hardness of dentinogenesis imperfecta and permanent dentin. METHODS: Totally 5 dentinogenesis imperfecta teeth were collected from 3 patients. At the same time, orthodontic teeth served as controls. The surface morphology, element content and micro-hardness of dentinogenesis imperfecta dentin and permanent dentin were measured by scanning electron microscope, X-ray energy chromatography and nano-indentation instrument. RESULTS AND CONCLUSION: The dentin structure of dentinogenesis imperfecta was loose, and the dentinal tubule structural was affected. The content of Na, Zn, S, Ca in dentinogenesis imperfecta was significantly greater than that of permanent dentin, but the content of Mg, P, Cl was significantly decreased than permanent dentin (P < 0. 01). The Ca/P ratio of dentinogenesis imperfecta was 1.76, which significantly higher than permanent dentin (P < 0.01). Microhardness test results showed micro-hardness of dentinogenesis imperfect was (1.18±0.18) GPa, which was significantly lower than that of normal permanent dentin (1.72±0.48) GPa (P < 0. 01). The dentinal tubule structural of dentinogenesis imperfecta was changed, and micro-hardness was reduced, all these may be related to the element content. Related Articles | Metrics

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Dynamic changes in immune function of active immunization-induced idiopathic thrombocytopenic purpura animal models Hu Hai-yan, Li Zhang-qiu, Yu Li, Yao Yao, Wang Guo-zheng, Sun Ming, Huang Rui, Zhang Mei-xia, Lu Zhi-gang 2010, 14 (24):  4462-4468.  doi: 10.3969/j.issn.1673-8225.2010.24.021 Abstract ( 170 )   PDF (537KB) ( 827 )   Save References | Related Articles | Metrics

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Effect of proteasome inhibitor MG-132 on myf-5 protein expression in rat denervated skeletal muscles Yan Shao-jun, Wang Dong, Zhang Wen-hui 2010, 14 (24):  4469-4472.  doi: 10.3969/j.issn.1673-8225.2010.24.022 Abstract ( 116 )   PDF (403KB) ( 543 )   Save BACKGROUND: MG-132 is a kind of proteasome inhibitor, which has delaying effect on atrophy of denervated skeletal muscle. OBJECTIVE: To detect the effect of proteasome inhibitor MG-132 on expression of myogenic regulatory factors Myf-5 gene of denervated skeletal muscle in rats. METHODS: Sprague-Dawley rats were randomly divided into three groups. Sham operation were made only for the rats in the control group (sciatic nerves were not cut off). Sciatic nerves were cut off more than 1-cm at the mid-level on their right lower limbs for the rats in the denervated and MG-132 intervention groups. Rats were sacrificed at 2, 7, and 28 days after operation. The expression of Myf-5 mRNA was detected by RT-PCR, and the changes of myf-5 protein were determined by Western blot. RESULTS AND CONCLUSION: Compared with the control group, the expressions of Myf-5 mRNA and protein were increased in the denervated and MG-132 intervention groups at 2, 7, and 28 days after operation (P < 0.01). The increasing of Myf-5 mRNA in the MG-132 intervention group was greater than that in the denervated group (P < 0.01). The results suggested that the proteasome inhibitor MG-132 can inhibit the ubiquitin-proteasome pathway to increase the expression of Myf-5 gene in order to play the role of slowing skeletal muscle atrophy. Related Articles | Metrics

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Expression of caspase-3/8 mRNA in hypertrophic scars Zhu Shao-ban, Liu Da-en, Li Hong-mian, Niu Hai-yan 2010, 14 (24):  4473-4476.  doi: 10.3969/j.issn.1673-8225.2010.24.023 Abstract ( 115 )   PDF (304KB) ( 291 )   Save BACKGROUND: Apoptosis participates in the dynamic process of cell component decease due to granulation tissues transforming into scar tissues, and Caspase family mediated apoptosis is the main pathway for apoptosis. Severe burns often reduces the formation of hypertrophic scars, Seriously affected in patients with body function and aesthetic, the treatment of hypertrophic scars are still exploring at the molecular level. OBJECTIVE: To explore the role of caspase-3 mRNA and caspase-8 mRNA in hypertrophic scars at the gene level. METHODS: Pathological scar tissue and normal skin tissue were obtained from 18 patients who received treatment at the Department of Burn and Plastic Surgery, the First Affiliated Hospital of Guangxi Medical University. Additional normal skin tissues served as controls. The expression of caspase-3 mRNA and caspase-8 mRNA in hypertrophic scars was tested by semi-quantitative reverse polymerase chain reaction method. RESULTS AND CONCLUSION: The expression rate of caspase-3 mRNA and caspase-8mRNA in hypertrophic scar were (8.31±1.47) and (7.12±1.24), respectively, which were significantly smaller than those of the normal skins (P < 0.05). The results indicated that caspase-3 mRNA and caspase-8 mRNA might play an important role in the process of wound healing and that the formation of hypertrophic scar might be due to the down-regulation of caspase-3 mRNA or (and) caspase-8 mRNA expression. Related Articles | Metrics

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Effect of telmisartan on adiponectin expression in adipocytes in vitro  Ning Guo-qing, Su Hua, Liu Chang-hui, Tan Xiao-jin 2010, 14 (24):  4477-4480.  doi: 10.3969/j.issn.1673-8225.2010.24.024 Abstract ( 135 )   PDF (342KB) ( 367 )   Save BACKGROUND: Adiponectin plays a regulatory role in insulin resistance and metabolic syndrome, and the synthesis of which is regulated by peroxisome proliferator-activated receptor γ (PPAR γ). Studies demonstrated that angiotensin receptor blocker  can affect the metabolism of adiponectin via activating PPAR γ, but the precise mechanism remains unclear. OBJECTIVE: To investigate the effect of telmisartan on the adiponectin expression in 3T3-L1 adipocytes in vitro, and to compare the results with candesartan. METHODS: 3T3-L1 adipocytes were cultured and differentiated in vitro. The differentiated 3T3-L1 adipocytes were cultured with vehicle or candesartan (10 μmol/L) or telmisartan (10 μmol/L). The expression level and secretary level of adiponectin were detected by RT-PCR, Western blot and ELISA. RESULTS AND CONCLUSION: Compared with the control and candisartan groups, telmisartan significantly increased the expression of adiponectin mRNA and protein in adipocytes (P < 0.05, P < 0.01), but there was no significant difference in secretion levels of adiponectin between groups (P > 0.05). The results demonstrated that telmisartan can increase the adiponectin expression in 3T3-L1 adipocytes, but can not increase the secretion by 3T3-L1 adipocytes. The promotive effect of telmisartan is superior to candesartan. Related Articles | Metrics

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Real-time monitoring of intracellular Ca 2+ concentration by flow cytometry Cao Yun-xin, Li Yu-rong, Yang An-gang 2010, 14 (24):  4481-4483.  doi: 10.3969/j.issn.1673-8225.2010.24.025 Abstract ( 316 )   PDF (275KB) ( 899 )   Save BACKGROUND: The concentration changes of Ca 2+ in cells is susceptible to various signal transduction pathways, thus, it is helpful to monitor changes in Ca 2+ for a better understanding of the priming, augmentation or inhibition of the functions of T-cells. OBJECTIVE: To detect dynamic changes in Ca 2+ using flow cytometry. METHODS: The fresh whole blood was obtained from human, and the lymphocytes were separated. CD3mAb and CD28mAb were added into the Fluo-3/AM-loaded cells, and the changes of Ca 2+ concentration were real-time monitored by flow cytometry. RESULTS AND CONCLUSION: The fluorescence value of Ca 2+ was stable and showed a baseline when the T lymphocytes at static status; after adding anti-CD3mAb and anti-CD28mAb, the T lymphocytes were activated, and the Ca 2+ concentration was quickly increased, lasted for 2-3 minutes, and gradually trended to be stale. The results revealed that flow cytometric analysis enabled us to trace dynamic movement of Ca 2+ using Fluo-3, which can be used as an effective method for monitoring Ca 2+ changes.  Related Articles | Metrics

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Research progress and clinical application of repairing bone defects using tissue engineered bone Chen Xin, Zhang Chun-lin 2010, 14 (24):  4486-4490.  doi: 10.3969/j.issn.1673-8225.2010.24.027 Abstract ( 212 )   PDF (385KB) ( 475 )   Save BACKGROUND: With the development of biotechnology, repairing bone defects using tissue engineered bone has become a hot spot. OBJECTIVE: To overview the research progress and clinical applications of repairing bone defects with tissue engineered bone. METHODS: The databases of PubMed, Springer Link, and Science Direct were retrieved for papers published from January 2000 to March 2009 with key words of “tissue engineering of bone, bone defect, and clinical applications”. The relevant papers concerning stem cell, scaffold material, animal model, vascularization during repairing bone defects with tissue engineered bone were collected. RESULTS AND CONCLUSION: Bone defect reparation has been a clinical challenge. The traditional autogenous bone transplantation was the gold standard for local bone defects, which had drawback of complication. It is an ideal technique that applying allogeneic bone scaffolds without number restrictions to repair bone defects, which had possibility of rejection reaction and infection. Therefore, the applications of tissue engineered bone are the tendency. The tissue engineered bone comprised 4 key elements: bone conductive scaffolds, releasing osteogenic growth factor, loaded osteogenic potential cells, as well as vascularization. Thus, the factors involved in osteanagenesis and its relations, as well as animal model studies should be evaluated during bone defects reparation. There have been shown positive roles and considerable development prospects of tissue engineered bone, but many issues have to be solved before the clinical applications. Related Articles | Metrics

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Current research situation of arthroidal cartilage injury and repair Gong Qing-bo 2010, 14 (24):  4491-4494.  doi: 10.3969/j.issn.1673-8225.2010.24.028 Abstract ( 180 )   PDF (360KB) ( 473 )   Save BACKGROUND: The cartilage injury is difficult to treat due to its limited self healing ability. OBJECTIVE: To review the current situation research of arthroidal cartilage injury and repair. METHODS: The databases of Chinese Biomedical Literature Database, CNKI, China Academic Journals Full-text Database, PubMed, EMbase were retrieved with key words of “arthroidal cartilage injury, arthroidal cartilage repair” for articles published from January 1990 to September 2009. The language was limited to Chinese and English. Papers concerning structure of articular cartilage, mechanism of articular cartilage injury, repairing the articular cartilage injury with articular cartilage transplantation, as well as using tissue engineering methods to regenerate articular cartilage were included. RESULTS AND CONCLUSION: The traditional cartilage repair methods mainly include subchondral drilling, grinding and micro fracture art, with purpose of mobilizing all the cell proliferation, which has the potential to proliferate cartilage cells, so as to achieve the objective reconstruction. However, the repairing tissues are often fibrocartilage, rather than transparent cartilage. Further degradation and the general problems such as the ossification also exist so general therapeutic effects were poor. In recent years, with the material science, engineering, and cell biology, physics and chemistry disciplines related to the development and cross, new kinds of articular cartilage repair defects, such as cartilage cells transplantation, tissue engineering and gene therapy, is gradually applied. Related Articles | Metrics

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Effects of loading/unloading exercise on bone mineral density in different populations Li Xiu-fen, Cui Ba-teer, Hao Bo-wen, Chen Yu-juan, Li Li 2010, 14 (24):  4495-4498.  doi: 10.3969/j.issn.1673-8225.2010.24.029 Abstract ( 137 )   PDF (291KB) ( 536 )   Save BACKGROUND: Human bone mineral density is affected by various factors, including exercise. OBJECTIVE: To summarize the effects of exercise on bone mineral density of different people and to provide the theory guide for scientific exercise. METHODS: Articles in accordance with inclusion and exclusion criteria were retrieved from CNKI and Medline database from 1985 to 2009. After data evaluation, in total 45 articles were included in the final analysis. The involved articles were summarized based on effects of exercise on bone mineral density of children, juveniles, middle-aged people and old people. RESULTS AND CONCLUSION: To teenagers, the mass of bone mineral density had important effects on peak value and adult osteoporosis. To middle-aged people, the decreased estrogen level would affect the role of exercise on bone mass. Aerobic exercise could maintain bone mineral density in a normal level and delay the senile osteoporosis. Exercise affected bone mineral density of different people. Various types of exercise have different effects and the loading exercise have more benefits than unloading exercise; and the effects of long-term exercise should be paid more attention. Related Articles | Metrics

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In situ hybridization in tissue engineering  Peng Qin, Wang Yuan-liang, Li Yu-xiao 2010, 14 (24):  4499-4502.  doi: 10.3969/j.issn.1673-8225.2010.24.030 Abstract ( 75 )   PDF (289KB) ( 536 )   Save BACKGROUND: In situ hybridization for detecting repaired tissue is an important tool to analyze tissue regeneration and repairing status. OBJECTIVE: To summarize the in situ hybridization in tissue engineering. METHODS: A computer-based online search was conducted in Pubmed for English language publications using the key words of “In situ hybridization, Tissue engineering, Insulin-like growth factor-I (IGF-I)” from January 1980 to July 2009. Relevant data were also searched in Chinese Journal Full-text Database with the same key words in Chinese from January 1995 to July 2009. Articles about mechanism and application of in situ hybridization were included. RESULTS AND CONCLUSION: In situ hybridization was an important instrument for the study of nucleic acid localization and the detection of gene expression, and it included molecular biology, cytogenetics, histochemistry and so on. In situ hybridization, in particular fluorescence in situ hybridization, might accurately and effectively detect the repaired tissue. The applications of in situ hybridization in tissue engineering have been achieved greatly, such as bone and cartilage. Many results present a good promise in biomedical and clinical medical fields in recent years. Besides, it is effective to in situ detection of insulin-like growth factor-I (IGF-I), which is closely related to growth and development of tissue. The emphasis of the future studies is about in situ detection of IGF-I in three-dimensional scaffold. Related Articles | Metrics

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Comparison between biomechanical measurement and traditional measurement    Yang Jian-xin, Ma Chao, Zhang Chun-lin 2010, 14 (24):  4503-4506.  doi: 10.3969/j.issn.1673-8225.2010.24.031 Abstract ( 114 )   PDF (348KB) ( 385 )   Save BACKGROUND: The ongoing development of biomechanics gradually separates the measuring techniques of biomechanics from traditional measuring methods in their nature and application in various areas of biomechanics. OBJECTIVE: A comparative study summarizing the latest research of biomechanical measurements and traditional measurements. METHODS: A computer-based online search was conducted in Science Direct database and Ei database with the key words of “biomechanics measurement” in English from January 1960 to October 2009. Also the relevant articles were searched in Chinese Journal Full-text Database and CBMdisc with key words of “biomechanics measurement” in Chinese from January 1994 to October 2009. In addition, several relevant monographs in print were used. The study belongs to all the following aspects: biomechanics measurement, biomechanics of sports, development status of biomechanics and measuring methods of biomechanics of sports.  RESULTS AND CONCLUSION: Biomechanical measurement and traditional measurement were compared in this research from both principles and practical applications. Biomechanical measurement was active, real-time, and limited, while it was highly correlated with medical science and biology. Besides studying the positive factors of the traditional measurement, this study was designed to try to apply them into the biomechanical measurement. Related Articles | Metrics

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Mechanical signaling pathways in skeletal muscle growth and adaptation Shi Reng-fei 2010, 14 (24):  4507-4511.  doi: 10.3969/j.issn.1673-8225.2010.24.032 Abstract ( 155 )   PDF (428KB) ( 578 )   Save BACKGROUND: Skeletal muscle, as a mechanical tissue, has a function of mechanical contraction, but the mechanisms by which mechanical signals are transduced to chemical signals that influence muscle growth and metabolism remain unidentified. OBJECTIVE: To understand the signaling pathways for mechanical stimulation and skeletal muscle adaptation, and to provide a theoretical basis for further study.  METHODS: The databases of PubMed and CNKI was retrieved by computer with key words of “skeletal muscle, mechanical stimulation, signal transduction” both in Chinese and English. The following literatures were included: ①Experimental papers with reliable arguments. ②Papers concerning skeletal muscle hypertrophy. ③Papers with definite standpoints and comprehensive analysis. The papers unrelated to this paper or repeatability study was excluded. RESULTS AND CONCLUSION: Several findings have suggested that mechanical stimuli can promote the growth and adaptation via Akt/mTOR pathway; under mechanical motion on skeletal muscle fiber, nerve impulse transfers the signals, and opens the Ca2+ ion channel, which can increase the calcineurin activity, activate NFAT into nucleus, and affecte responding target gene, ultimately, result in the adaptation of skeletal muscle. It demonstrated that muscle releases insulin-like growth factors 1 on mechanical stimulation, and through signaling pathways influence muscle growth and adaptation. On the other hand, mechanical signaling via the calcineurin/nuclear factor of activated T-cell pathway has been shown to have a powerful influence on muscle phenotype and muscle mass. The mechanisms concerning how to change the mechanical signals into in vivo chemical signals pathways need exploration.  Related Articles | Metrics

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Effects of MG29 protein on excitation-contraction coupling in senescent muscles  Qu Hui-fang, Pan Wen-zhao, Yan Wan-jun 2010, 14 (24):  4512-4515.  doi: 10.3969/j.issn.1673-8225.2010.24.033 Abstract ( 157 )   PDF (323KB) ( 333 )   Save BACKGROUND: Excitation-contraction coupling is a basic functional feature of nerve-muscle, to some extent, which closely associates with functional deterioration of senescent muscles. Researchers have made obviously progress in this field recently. OBJECTIVE: To summarize the mechanism of the change of Ca2+ in senescent muscle cells and the effects of MG29 protein on Ca2+ changes and excitation-contraction coupling, and to provide reference for studying functional deterioration via reviewing related literatures.  METHODS: Papers related to mechanism of the change of Ca2+ in senescent muscle cells and correlation between MG29 protein and excitation-contraction coupling published in CNKI (2000-01/2009-11) in Chinese and in PubMed (2009-01/2009-12) in English were searched by computer based on the keywords of “excitation-contraction, muscle, muscle cell, senescence, resistance training and MG29”. Obsolete, repetitive and credibility-lacking literatures were excluded. RESULTS AND CONCLUSION: Totally 43 papers were retrieved. According to the inclusive and exclusive criteria, 9 papers were excluded and 34 documents were included in the final analysis. The results showed that MG29 plays an important function in excitation-contraction coupling and decreases with age increasing. Therefore, MG29 is speculated as a key factor of muscle senescence. Importantly, recent researches show that MG29 could be increased by resistance training. Related Articles | Metrics

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Application of genetic and transgenic technology in repairing chronic wounds Zhang Wang, Lu Wei 2010, 14 (24):  4516-4519.  doi: 10.3969/j.issn.1673-8225.2010.24.034 Abstract ( 150 )   PDF (366KB) ( 361 )   Save BACKGROUND: Refractory wound is difficult to treat using traditional methods in the clinic. Studies have shown that growth factor plays an important role in wound healing, and transgenic therapy is emerging to treat refractory wound.  OBJECTIVE: To review the research status and progress of treating refractory wound by summarizing literatures concerning gene therapy. METHODS: PubMed database was searched by the first author using key words of “gene therapy, burns, chronic wounds” for English papers published between January 1999 and December 2008. Documents with regard to refractory wound and gene therapy were included. Totally 33 literatures were analyzed.  RESULTS AND CONCLUSION: According to sources of transfected cells, the transgenic technology can be divided into in vivo gene transfection and in vitro gene transfection. The in vivo gene transfection processes greater superiority than other methods. Though it can not be spread in clinical application, transgenic technology owns extensive prospect. Related Articles | Metrics

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Theoretical basis and effect of treating soft-tissue disorders using acupotomology      Zhang Yi, Guo Chang-qing 2010, 14 (24):  4520-4523.  doi: 10.3969/j.issn.1673-8225.2010.24.035 Abstract ( 128 )   PDF (346KB) ( 455 )   Save The theoretical basis and effect of acupotomology treatment were analyzed by summarizing existing findings. Soft tissue plays an important role in the body. The fibrosis, hyperplasia, and hypertrophy of soft tissues can cause the change of soft tissue in relative motion disorder, tension increase, length shorten, lacuna pressure increase. All the changes can not only aggravate soft tissue lesion, but can also act on bone, joint, nerve, blood vessel and other organs or tissues to participate pathologic process of many diseases. The main therapeutical effect of acupotomology is soft tissue dissolution and destruction. Acupotomology is a kind of percutaneous minimally invasive soft tissue release treatment. By puncture, small area carve and blunt dissection in soft tissue, it can release nerve, blood vessel and osteoarticular from the jeopardize that caused by abnormal soft tissue. Besides soft tissue release, acupotomology has the similar effect to acupuncture. Acupuncture can two-ways regulate human organs function by neuroendocrine-immune modulation, which is known as integral regulation. Related Articles | Metrics

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Associations of transforming growth factor beta 1-509C/T site gene polymorphism with blood stasis syndrome of lumbar intervertebral disc protrusion and intervertebral disc degeneration Xu Jian-wen, Zhong Yuan-ming, Ji Jing, Li Zhi-fei, Chen Wen-fu, Su Jie-han, Huang Zeng-chao, Li Gui-huan 2010, 14 (24):  4524-4527.  doi: 10.3969/j.issn.1673-8225.2010.24.036 Abstract ( 153 )   PDF (306KB) ( 391 )   Save BACKGROUND: Blood stasis syndrome is common in lumbar intervertebral disk protrusion (LIDP) in traditional Chinese medical clinical practice, which lacks of ideal preventing and treating methods. Previous studies have reported that transforming growth factor β1 (TGF-β1) -509C/T site gene polymorphism may correlate to intervertebral disc degeneration (IDD).  OBJECTIVE: To investigate the relationship among the blood stasis syndrome in LIDP, IDD and TGF-β1-509C/T site gene polymorphisms. METHODS: Totally 120 LIDP patients who received treatments in Department of Orthopedics of First Affiliated Hospital of Guangxi Traditional Chinese Medical College from January 2008 to December 2009 were selected, including 60 blood stasis syndrome and 60 non-blood stasis syndrome. The TGF-β1-509C/T site gene polymorphisms were detected and the IDD indexes were observed. The relationships between TGF-β1-509C/T site gene polymorphisms and the blood stasis syndrome of LIDP were analyzed by logistic regression Forward methods. The grade of intervertebral disc degeneration consisted of mild, middle and severe according to nuclear magnetic resonance. RESULTS AND CONCLUSION: The degenerative degree of lumbar intervertebral disk with TGF-β1 gene-509 CT was higher than CC and TT in LIDP patients, the blood stasis syndrome group was severer than the non-blood stasis syndrome group (P < 0.05). It was showed that the risk of syndrome of blood stasis of LIDP in patients with severe IDD was 1.818 folds of that in patients with mild and middle IDD (OR =1.818, 95%CI: 1.275-2.931, P < 0.05), and the risk further increased 2.038 folds after accumulating with TGF-β1-509C/T site gene polymorphisms (OR = 2.038, 95%CI:1.379-3.423, P < 0.05). Severe IDD may be one of the risk factors of syndrome of blood stasis of LIDP in Han ethnic population from Guangxi. Patients with TGF-β1 gene-509 adiponectin CT genotype and combined with severe IDD are susceptible to syndrome of blood stasis of LIDP. Related Articles | Metrics

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Prevalence and risk factors of pre-hypertension in urban and rural population of Beijing  Hu Bo, Li Wei, Liu Bing, Chen Tao, Sun Yi 2010, 14 (24):  4528-4532.  doi: 10.3969/j.issn.1673-8225.2010.24.037 Abstract ( 104 )   PDF (356KB) ( 354 )   Save BACKGROUND: Pre-hypertension investigation plays an important role in preventing, reducing and delaying hypertension. However, there are few reports concerning the pre-hypertension in national large-scale investigations. OBJECTIVE: To compare the prevalence and risk factors of pre-hypertension in urban and rural population of Beijing, to explore the effects of urbanization on the pre-hypertension, in addition, to provide guidance for disease prevention and control.  METHODS: A cluster sampling was used to establish a study population of inhabitants aged 40-65. The definition of pre-hypertension was determined by National Revision Committee of the Guidelines of Hypertension Prevention and Control in 2005. Logistic regression model was used to find the risk factors of pre-hypertension. RESULTS AND CONCLUSION: A cohort of 3 268 middle-aged participants were enrolled in the investigation. The prevalence of hypertension in urban was lower than in rural (P < 0.05). Multivariate logistic analysis revealed that, in males, the 0.1 unit increase in waist-hip ratio led to 1.411 increase in risk factor (95% CI: 1.031-1.931), and the current drinking people suffered 0.648 risk factors than that never drinking (95% CI: 0.437-0.961). In females, the 0.1 unit increase in waist-hip ratio resulted in 1.489 increase in risk factor (95% CI: 1.006-2.203), and 1 unit increase in triglyceride caused 1.194 increase in risk factor (95% CI: 1.000-1.426). Accordingly, obesity is a major risk factor for pre-hypertension.  Related Articles | Metrics

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Polymorphism of vitamin D receptor and expression of CYP3A4 gene in intestinal tract: Does the distribution of vitamin D receptor mutation have difference in population? Li Bao-qun, Hao Zhi-min, Wan Li-juan 2010, 14 (24):  4533-4537.  doi: 10.3969/j.issn.1673-8225.2010.24.038 Abstract ( 152 )   PDF (385KB) ( 454 )   Save BACKGROUND: Studies have demonstrated that vitamin D receptor (VDR) induces CYP3A4 gene transcription and expression in tract, and the abnormity of VDR will change the quantity and quality of its protein and cause alteration of target genes transcription and expression or function in the downstream. OBJECTIVE: To identify the effects of VDR Fok Ⅰ mutation on the transcription of CYP3A4 in intestinal tract HT-29 cells. METHODS: Hepatic tissues were obtained from hepatolithus patients and constructed pcDNA3.1(-)B-myc/his h VDR eukaryotic expression vectors (wild type and Fok Ⅰ mutant). The HT-29 cells were transfected by using cell transfection technique and dual-luciferase report gene analytical system and treated by different concentrations of 1,25(OH)2VD3 (1, 10, and 100 nmol/L). The transcription of CYP3A4 in cultured HT-29 cells was observed. RESULTS AND CONCLUSION: In transiently transfected HT-29 cells, the CYP3A4 luciferase activity of three concentrations (1, 10, 100 nmol/L) was increased respectively after 1,25(OH)2VD3 was added to cells for 24 hours, and there were significant differences between 1,25(OH)2VD3 group and the vehicle control group (P < 0.05), the luciferase activity from mutant VDR constructs was a little greater than wild VDR constructs, but there was no significant difference between the two VDR forms (P > 0.05). VDR Fok Ⅰ mutant could not significantly change the regulational capacity of wild type VDR on CYP3A4. The results revealed that vitamin D induces CYP3A4 expression via VDR. However, compared with wild type, the VDR Fok I mutant has no obvious difference on CYP3A4 expression. Related Articles | Metrics

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Somatotype of 525 Han brain workers in Dongying region of Shandong province Liu Yi, Yuan Xun-yi, Han Ying, Li Song, Ji Ai-yu 2010, 14 (24):  4538-4542.  doi: 10.3969/j.issn.1673-8225.2010.24.039 Abstract ( 122 )   PDF (324KB) ( 425 )   Save BACKGROUND: Synthetical analysis of human somatotype is late in China, and there are relatively less documents, especially for certain occupational groups. OBJECTIVE: To determine the somatotype components and somatotype characteristics of the Han brain workers of Dongying region of Shandong province, and to analyze the difference caused by gender and age.  METHODS: The Heath-Carter anthropometric somatotyping method was used to measure 525 healthy participants including 264 male and 261 female brain workers selected from Dongying. They were divided into groups by gender and age. The somatotype components were calculated and compared by t-test and analysis of variance to test differences among the somatotype means and the somatotype attitudinal distance (SAD).  RESULTS AND CONCLUSION: The somatotype means and the somatotype distributions and somatotype components of the males was 5.3-4.3-1.7, the ratio of height and body mass was 41.25 and the rate of body fat was 20.41%. The somatotype means and the somatotype distributions and somatotype components of the females were 6.4-3.7-1.1, the ratio of height and body mass was 40.12 and the rate of body fat was 28.75%. There were no significant differences in terms of extrinsic factor and ratio of height and body mass between the sexes (P > 0.05). The mesomorphic means of the males were greater than the females, while the endomorphic means of the females were greater than males (P < 0.01). For the males, the mean somatotypes were mainly mesomorphic, displaying developed skeleton and muscles as well as strong body. For the females, the mean somatotypes were mainly endomorphic, displaying plump body, thick subcutaneous fact and high body fat. The somatotypes means and the somatotype distributions and somatotype components vary with gender and age. The endomorphy of the Han brain workers of Dongying region is relatively greater. Related Articles | Metrics

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Analysis of body composition of excellent long jumpers in China   Hou Yu-lu, Peng Hong 2010, 14 (24):  4543-4546.  doi: 10.3969/j.issn.1673-8225.2010.24.040 Abstract ( 267 )   PDF (196KB) ( 678 )   Save BACKGROUND: Body composition of certain excellent athletes has been reported, however, few studies concerning the body composition of excellent long jumpers in China. OBJECTIVE: To explore the body composition and distribution of excellent long jumpers in China through determining body composition using biological resistance method. METHODS: A total of 67 long jumpers who participated in the 2005 National Athletics Championships and 2004 National University Athletics Meeting were included, 60 track-and-field specialized students and 30 university students were selected as controls. The body composition, including body fat rate, lean body mass, basal metabolic rate (BMR) and total water content were measured using “Bios pace” component analyzer with eight electrodes method. The age, gender, morphological index, and body composition were compared between the two groups. RESULTS AND CONCLUSION: The distribution features of excellent long jumpers in China were as follows: low body fat rates, body weight, and free fat body weight. The body fat rates were gradually decreased and lean body mass increased with increasing athletic levels. There were significant differences between the male and female long jumpers (P < 0.01). The lean body mass, lean body mass/height and vital capacity had positive correlation with standing long jump, standing triple jump, deep squat and special achievement, but were negatively related to 30-m running. Compared with female, the correlation coefficient of the male with lean weight was greater in male. The results are corresponding to special achievement, thus, it can provide basis for selecting and training of athletes. Related Articles | Metrics

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Effect of extracorporeal shock wave on proliferation, differentiation, adhesion and migration of rat osteoblasts in vitro Huang Zhong-lian, Hu Jun, Yu Meng-lei, Lu Zhi-jun 2010, 14 (24):  4547-4552.  doi: 10.3969/j.issn.1673-8225.2010.24.041 Abstract ( 134 )   PDF (394KB) ( 480 )   Save BACKGROUND: Extracorporeal shock wave therapy is indicated as an effective method for treatment of delayed fracture healing or nonunion. Osteoblasts plays an important role in this process. OBJECTIVE: To investigate the function of osteoblasts in the process of extracorporeal shock wave promoting fractures healing, and to provide theoretical support for improving shock wave therapy on fracture healing. METHODS: Primary cultured ostsoblasts were isolated from newborn SD rat calvaria and randomly divided into two groups, shock wave and control. Treated by different energies of extracorporeal shock wave, cells were incubated onto 96-well culture plate. An optimal dose of extracorporeal shock wave was selected according to survival and proliferation of osteoblasts. The osteoblasts treated by optimal energy of extracorporeal shock wave were cultured and harvested for the analysis of alkaline phosphatase by calcium cobolt stain, cell survival by CCK-8 Kit, alkaline phosphatase expression by AKP kit, mineralized nodules by Alizarin red staining, integrin β1 and β1 mRNA expressions by flow cytometry and RT-PCR, cell migration by wound healing assay. RESULTS AND CONCLUSION: The optimal energy of extracorporeal shock wave treating primary cultured osteoblasts was   10 kV (500 impulses). Following extracorporeal shock wave therapy, the cell proliferation, alkaline phosphatase activity, cell mineralization, rates of cell adhesion, as well as β1 integrin and its mRNA expressions were increased as compared with those in control group (P < 0.01). Further distance of cell migration was found in extracorporeal shock wave group (P < 0.05). The results showed that the optimal energy of extracorporeal shock wave could promote the proliferation, differentiation, adhesion and migration of osteoblasts in vitro, and β1 integrin may play an important role in the process of cell adhesion and migration. Related Articles | Metrics

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Construction and identification of pIRES2-EGFP-hBMP-2 bicistronic eukaryotic expression vector Wang Jian-qiang, Zhang Hai-ning, Ding Chang-rong, Wang Ying-zhen 2010, 14 (24):  4553-4556.  doi: 10.3969/j.issn.1673-8225.2010.24.042 Abstract ( 111 )   PDF (278KB) ( 455 )   Save BACKGROUND: Bone morphogenetic protein (BMP) and its derived scaffold or vector here been widely used in experiment and gradually applied in the clinic. However, the tracing method challenges the therapeutic effect. The discovery and application of enhanced green fluorescent protein (EGFP) can solve this problem. OBJECTIVE: To construct the bicistronic eukaryotic vector pIRES2-EGFP-hBMP-2 and to observe its expression in human embryo kidney (HEK) 293 cells. METHODS: hMBP-2 gene was extracted from human osteosarcoma by RT-PCR and inserted into PMD18-T vector. Following the DNA sequence verification, it was then sub-cloned into the eukaryotic vector pIRES2-EGFP. After restriction enzyme analysis, the pIRES2-EGFP-BMP-2 was transfected into HEK 293 cells. Then its expression was observed using fluorescence microscopy and Western blot. RESULTS AND CONCLUSION: hBMP-2 gene was amplified and the eukaryotic vector pIRES2-EGFP-hBMP-2 was constructed successfully. At 48 hours after tranfection of pIRES2-EGFP-hBMP-2 into HEK 239 cells, approximately 30% of cells emitted green fluorescence under a fluorescence microscope. Western blot results demonstrated that there was a specific BMP strap at Mr46×106 side, which indicated that the target gene could be expressed in constructed pIRES2-EGFP-hBMP-2 vector. Related Articles | Metrics

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Chondron: A basic microanatomical unit in articular cartilage Duan Wang-ping, Sun Zhen-wei, Wei Xiao-chun 2010, 14 (24):  4557-4560.  doi: 10.3969/j.issn.1673-8225.2010.24.043 Abstract ( 287 )   Save BACKGROUND: Chondron is a basic microanatomical unit of articular cartilage. Increasing evidence suggests that the pericellular matrix (PCM) is a distinct functional compartment in articular cartilage, influencing the metabolic, micromechanical environment, and degeneration of chondrocytes. But the precise functions and action mechanism need further investigation. OBJECTIVE: To review the literature pertinent to the morphology, function, isolation of the chondron in articular cartilage, and its degenerative events during the progression of osteoarthritis (OA). METHODS: This review summarized the articles published in the PubMed database before July 2009. In addition, recent data and figure of our laboratory on the morphology and biomechanics of chondron and chondrocyte were supplemented. RESULTS AND CONCLUSION: The PCM is primarily characterized by the presence of type Ⅵ collagen, and these components are widespread in the expansive extracellular matrix (ECM) in newborns, while in mature the components are mainly localized to a narrow pericellular zone. The three-dimensional morphology of chondron has been recently quantified in situ with fluorescence confocal microscopy, and the mechanical properties of the isolated individual chondrons and their PCM are measured using the micropipette technique and atomic force microscopy. More studies have shown that the presence of the PCM in chondrons has a profound influence on chondrocyte gene expression. At the same time, structural, histochemical and biomechanical studies indicate the chondron and their PCM may undergo degenerative processes with osteoarthritis, similar to those occurring in the ECM. Although the precise function of the PCM is unknown, increasing evidence in vivo or in vitro suggests that the PCM is a basic microanatomical unit in articular cartilage, influencing the metabolic and micromechanical environment of chondrocytes. Related Articles | Metrics