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12 March 2010, Volume 14 Issue 11 Previous Issue    Next Issue

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Effect of Weilingxian on proliferation of rabbit knee articular chondrocytes cultured in vitro and mRNA expression of transforming growth factor-beta 1 Ma Yong, Zhang Yun-shen, Chen Jin-fei, Jin Xiang, Wang Pei-min, Wang Jian-wei, Xu Jian-an 2010, 14 (11):  1901-1906.  doi: 10.3969/j.issn.1673-8225.2010.11.001 Abstract ( 324 )   PDF (535KB) ( 672 )   Save BACKGROUND: Studies have shown that Weilingxian can maintain and promote the synthesis of proteoglycan and collagen Ⅱ of chondrocyte, and protect articular cartilage and postpone the development of osteoarthritis by inhibiting the level of interleukin-1 (IL-1) possibly. OBJECTIVE: Based on the previous studies, to observe the effect of Weilingxian on the proliferation of rabbit knee articular chondrocyte and transforming growth factor-β1 mRNA expression, and then to explore the role and possible mechanism of Weilingxian in the treatment of osteoarthritis. METHODS: Knee cartilage was shredded after harvested from New Zealand white rabbits under sterile conditions, and chondrocytes were isolated and cultured by the way of enzymatic digestion. After identifying by toluidine blue staining, the third-passage cells in the logarithmic growth phase were cultured in vitro and randomly divided into two groups after adherence. The experimental groups were cultured in DMEM with 0.01, 0.05, 0.1, 0.5, and 1.0 mg/mL Weilingxian, while the control group was given with normal medium alone. Chondrocytes morphology was observed under an inverted phase contrast microscope, and the phenotype was identified by toluidine blue staining; Methyl Thiazolyl Tetrazolium (MTT) assay method was adopted to observe the influence of Weilingxian with different concentrations on the proliferation of chondrocytes, and anti-transcription-polymerase chain-type reaction (RT-PCR) was used to assay the expression changes of transforming growth factor-β1 mRNA. RESULTS AND CONCLUSION: Primary cultured chondrocyte was round-shaped, and most of it adhered after 24 hours, the appearance was polygonal and irregular-shaped; after passage, cell growth was faster than before, the typical appearance was slabstone-like; long spindle-shaped chondrocytes appeared after four generations; after six generations, most cells showed long spindle-shaped fibroblast-like appearance, the rate of growth also slowed down. Extracellular matrix of chondrocytes was stained to be blue by toluidine blue staining, and the nucleus was dark blue. Different concentrations of Weilingxian could promote the proliferation of chondrocytes, effect of 0.5 mg/mL group was significantly, and the peak of proliferation was on the third day. 0.05, 0.1, 0.5, and 1.0 mg/mL Weilingxian group could promote the expression of transforming growth factor-β1 mRNA, and there was no significant difference between four groups (P > 0.05), but the peak was at 0.5 mg/mL group. Weilingxian can promote proliferation of chondrocyte and transforming growth factor-β1 mRNA expression, and these may be one of the possible mechanisms that Weilingxian can work in the treatment of osteoarthritis. Related Articles | Metrics

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Tensile mechanical properties of human tracheal cartilage Chen Shu, Guo Xi-ping, Lü Ya-ping, Ma Hong-shun 2010, 14 (11):  1907-1910.  doi: 10.3969/j.issn.1673-8225.2010.11.002 Abstract ( 678 )   PDF (350KB) ( 748 )   Save BACKGROUND: Tracheal injury suture and the development of a new tracheal prosthesis all need to understand the tensile mechanical properties of tracheal cartilage, so as to repair and rebuild tracheal function. Previous researches on the biomechanics of artificial trachea are many reported, while the biomechanics of human tracheal cartilage is reported less. OBJECTIVE: To investigate the mechanical properties of tracheal cartilage using one-dimensional tensile test method. METHODS: Two fresh cadaver specimens of normal human trachea, with the informed consents of their families, were involved. The specimens were thaw at room temperature, and then tracheal cartilage specimens were cut using scalpel into 20 samples at the length of 25 mm, width 5 mm, thickness 1.8-2.2 mm. The 20 tracheal cartilage samples were subjected to one-dimensional tensile test with Shimadzu electronic universal testing machine Japan, at the tensile test speed of 5 mm/min. The tensile maximum load, maximum displacement, maximum stress, maximum strain, elastic modulus and stress-strain curve of the specimens were observed. RESULTS AND CONCLUSION: The maximum load of human cadaver tracheal cartilage was (60.946±10.377) N, maximum displacement was (1.973±0.159) mm, maximum stress was (6.229±1.125) MPa, maximum strain was (32.825±2.776)%. Tracheal cartilage stress-strain curves was changed along with the index, the initial low slope of the curve was due to the direction of imposed tension was similar with the arrangement of collagen structure, the steep slope represented the tensile strength of collagen. To describe the tracheal cartilage stress-strain relationship in a one-dimensional tension, 15 stress-strain data of the tracheal cartilage experimental data adopted polynomial by the least square fitting method, to obtain stress (δ) - strain (ε) relationship formula:  . It is indicated that tracheal cartilage has a strong capacity to bear load and resist deformation, reflecting their viscoelastic mechanical properties of both flexibility and viscosity, supporting that the mechanical properties of cartilage is positively related to cartilage collagen content. Related Articles | Metrics

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Culture of rabbit periosteal osteoblasts using modified enzymatic digestion combined with explant method Zhao Bing-jing, Wang Zhi-ying 2010, 14 (11):  1911-1914.  doi: 10.3969/j.issn.1673-8225.2010.11.003 Abstract ( 403 )   PDF (526KB) ( 513 )   Save BACKGROUND: Periosteal osteoblasts possess strong reproductive activity, as well as osteoblastic differentiation potential, which is an ideal seed cell if can shorten the culture time. OBJECTIVE: Modified enzymatic digestion was used to culture rabbits’ osteoblasts, and to study the adherence and proliferation of osteoblasts on the surface of sandblasting titanium. METHODS: Periostea were harvested from the theanteromedial surface of the proximal tibia of male, Japanese white rabbits, and cultured as follow: ①Routine method: Digested with 0.25% trypsinase at 37 ℃ for 30 minutes, followed by digestion with 0.1% type I collagenase at 37 ℃ for 30 minutes, vibration, removed trypsinase and dried. After 2 hours, DMEM containing 15% fetal bovine serums were added. ②Modified method: 30 minutes culture of type I collagenase was prolonged to 1 hour. The osteoblasts were identified by alkaline phosphatase staining and calcium node staining. The adherence and proliferation of osteoblasts cultured on sandblasting surface were measured by scanning electron microscopy and MTT. RESULTS AND CONCLUSION: Five days after culture, the periosteal osteoblasts crawled out from tissues, gathered as monolayer with triangle or polygon at after 25 days of modified culture. After 1 month of culture, superposition growth of calcium nodus appeared. The cultured cells possessed the morphological characteristic and biological behavior of osteoblasts, which were positive to alkaline phosphorase and calcium node staining. The time of cells cultured with routine method covered flask delayed 12 days than modified method. The osteoblasts were inseted into sandblasting titatium with pseudopodium. However, the adherence and proliferation of osteoblasts cultured on sandblasting surface had no obviously difference between two culture methods. The results suggested that modified enzymatic digestion can shorten the culture time without effect on adherence and proliferation of osteoblasts.  Related Articles | Metrics

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Inhibition of peroxiredoxin Ⅱ on human intervertebral disc cells cultured in vitro Liu Yu-lin, Zhou Chu-song, Chen Zhong, Luo Ping, Lin Li-jun 2010, 14 (11):  1915-1918.  doi: 10.3969/j.issn.1673-8225.2010.11.004 Abstract ( 330 )   PDF (433KB) ( 468 )   Save BACKGROUND: Intervertebral disc degeneration can reduce nucleus pulposus cells, and peroxiredoxin Ⅱ involved in the regulation of resist oxidation damage, cell division, differentiation, signal transduction and apoptosis. Peroxiredoxin Ⅱ has promotive effect on intervertebral disc degeneration, whereas the mechanism remains poorly understood.     OBJECTIVE: To observe the effects of peroxiredoxin Ⅱ on human intevertebral disc cells activity and type Ⅱ collagen synthesis in vitro. METHODS: Human degenerated human lumbar disc cells were cultured in vitro, and assigned into the control and peroxidase Ⅱ groups. Peroxidase Ⅱ with doses of 10, 100 and 1 000 ng/L were added into the peroxidase Ⅱ groups. The cells were identified by immunohistochemical staining, and the cell proliferation was detected using cck-8 kit. Cell supernatant was collected at days 3 and 7 after operation, and the expression of type Ⅱ collagen was measured by double-antibody sandwich enzyme-linked immunosorbent assay. RESULTS AND CONCLUSION: In vitro cultured human degenerative lumbar intervertebral disc nucleus pulposus cells by adding peroxidase increased with the dose-Ⅱ, the disc nucleus pulposus cells of the volume and type Ⅱ collagen synthesis gradually reduced (P < 0.01). Tips peroxidase Ⅱ on the intervertebral disc nucleus pulposus cells, the number and type Ⅱ collagen synthesis significantly inhibited in a dose-dependent manner. Thus speculated that peroxidase Ⅱ on the nucleus pulposus cells in vitro may lead to disc degeneration as a precipitating factor. Related Articles | Metrics

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Culture and identification of rabbit aortic endothelial cells: Modified isolation and culture of endothelial cells Gao Hang, Guan Chun-yan 2010, 14 (11):  1919-1922.  doi: 10.3969/j.issn.1673-8225.2010.11.005 Abstract ( 367 )   PDF (365KB) ( 585 )   Save BACKGROUND: Although the methods of mechanical scrapping, tissue block transplantation, and enzyme digestion to culture endothelial cells have been quite mature, it has been constantly updated. OBJECTIVE: To explore the culture and identification of rabbit aortic endothelial cells. METHODS: Aorta was derived from one-week-old New Zealand rabbit. Following removing the adventitia, the endothelium was digested in the mixture of 2 g/L type Ⅰ collagen, 2 g/L type Ⅱ collagen, 2 g/L type Ⅳ collagen, and 2 g/L type Ⅴ collagen (1:1:1:1) for 20 minutes, and the digestion was stopped with culture medium (1:1). Cells in the endothelium were lightly scraped to make cell suspension, which was centrifuged to precipitate cells with DMEM culture medium containing 20% fetal bovine serum, 1 μg/L VEGF, 2 μg/L bFGF, and 6 U/L gentamicin. The cells were then blown into single cells and the culture liquid was changed every 48 hours. The cells were passaged according to the ratio of 1:2. Inverted phase contrast microscope was used to observe cell culture; immunohistochemistry and immunofluorescence were used to determine Ⅷ-related antigen; electron microscope was used to detect Weibel-Paladed body. RESULTS AND CONCLUSION: VIII-related antigen and electron microscopy confirmed that the primary and 5-passage endothelial cells were successfully cultured, suggesting that rabbit aortic endothelial cells were cultured into cell line. VIII-related antigen and electron microscopy determined that a combination with W-P body was a good method to determine endothelial cells. Related Articles | Metrics

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Effect of pentoxifylline on proliferation of human keloid fibroblasts Feng Hong-xia, Xin Yan, Hao Yu-qin, Kang Chun-yi 2010, 14 (11):  1923-1926.  doi: 10.3969/j.issn.1673-8225.2010.11.006 Abstract ( 453 )   PDF (380KB) ( 522 )   Save BACKGROUND: In recent years, pentoxifylline has been found to have a wide range of anti-fibrosis capacity. However, there are few studies explore the suppress effect of pentoxifylline on fibroblasts in human keloid, and the maximum inhibitory concentration remains poorly understood. OBJECTIVE: To study the effect of pentoxifylline on proliferation activity of human keloid fibroblasts and to select the maximum inhibitory concentration. METHODS: Human keloid fibroblasts were used as original cells, passaged till the 5th to the 8th generations, and then divided into the experimental and control groups. Pentoxifylline with concentrations of 0.1, 0.25, 0.5, 1.0, 2.0 and 3.0 g/L were added to the experimental group. The effects of different concentrations of pentoxifylline on proliferation of keloid fibroblasts were detected by MTT chromometry. RESULT AND CONCLUSION: Compared with the control group, the inhibitory effect of pentoxifylline on the proliferation of keloid fibroblasts was more evident in the experimental group (P < 0.05). The inhibition rates of pentoxifylline on proliferation of keloid fibroblasts showed apparently time- and dose-effect relationships within the concentration of 0.1- 2.0 g/L, which presented a greatest level at 96 hours after culture. The maximum inhibitory rate was 53.37%, and the concentration was 2.0 g/L in the experimental group. Consequently, pentoxifylline plays a notable inhibitory role in the proliferation of human keloid fibroblasts with concentration of 2.0 g/L at 96 hours after culture.  Related Articles | Metrics

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Expressions of Smad3 and transforming growth factor beta 1 in keloid, hypertrophic scars and normal skins: A 48: 40: 40 sample pathological detection Pang Jiu-ling, Ma Zheng, Liu Jun, Liu Ai-dong 2010, 14 (11):  1927-1930.  doi: 10.3969/j.issn.1673-8225.2010.11.007 Abstract ( 368 )   PDF (356KB) ( 572 )   Save OBJECTIVE: Study regarding Smad3/transforming growth factor-β1 (TGF-β1) signal transduction in pathological scars mainly focus on in vitro cultured fibroblasts, however, the correlation study was rare on keloid. The aim of this study is to detect the expressions of Smad3 and TGF-β1, and investigate their relationship in pathogenesis and development of pathological scars. METHODS: Experimental samples were obtained from the patients, who underwent burn and plastic surgery at the Department of Burn and Plastic Surgery, Workers’ Hospital of Tangshan, Hebei Medical University, from June 2004 to June 2008, including 48 patients with Keloid, aged 16-52 years, and 40 patients with hypertrophic scars aged 18-56 years. Normal skins from additional 40 cases were served as controls. The expressions of Smad3 and TGF-β1 protein in keloid, hypertrophic scars and normal skin were examined by flow cytometry. RESULTS: The expression of Smad3 and TGF-β1 were obviously greater in the experimental group than that of the control group (P < 0.05), but the difference between keloid and hypertrophic scars had no significance (P > 0.05). There was a positive correlation between Smad3 and TGF-β1 in keloid and hypertrophic scars (r=0.489 2, P=0.000 4; r=0.471 0, P=0.002 2). No notable correlation was found between Smad3 and TGF-β1 in normal skins (P=0.471 4). CONCLUSION: The expressions of Smad3 and TGF-β1 are up-regulated, and the synergism of Smad3 and TGF-β1 may promote the development in pathological scars. Related Articles | Metrics

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Role of preoperative hormone therapy in localised prostate cancer: Meta-analysis based on six randomized controlled trials Li Tao, Yang Ke-hu, Tian Jin-hui, Chen Yi-rong, Li Chao-bin, Guo Bai-hong, Li Guo-ping, Guo Qing-hua 2010, 14 (11):  1931-1934.  doi: 10.3969/j.issn.1673-8225.2010.11.008 Abstract ( 337 )   PDF (348KB) ( 462 )   Save BACKGROUND: Previous research has demonstrated that preoperative hormone therapy relieved clinical and pathological stages of prostatic carcinoma patients, reduced positive ratio of incisal margin; however, it did not increase disease-free survival rate. Worth of preoperative hormone therapy for prostatic carcinoma remains unclear. OBJECTIVE: To assess the role of preoperative hormone therapy in localised prostate cancer. METHODS: Articles were searched from PubMed, Embase, Cochrane Library (No. 4, 2009), Chinese biomedical literature database, Chinese Scientific Journals full-text database, and Chinese Journal full-text database (published before October, 2009). Randomized controlled trials which were diagnosed as localised prostate cancer using pathology and cytology were included. The sex and nationality were not limited, and patients did not have severe heart and lung diseases. Prostatic carcinoma which occurred later or repeatedly was excluded. Otherwise, randomized controlled trails which were coincidence with the inclusion criteria were also included. Review Manager 5.0 published by Cochrane was used for statistical analysis. Overall survival rate, disease-free survival rate, positive surgical margin rate, positive lymph node rate, and seminal vesicle invasion rate were evaluated. RESULTS AND CONCLUSION: Six randomized controlled trials totaling 1 027 participants were included. The result of meta analysis showed that there was no significant difference in overall survival rate [RR=0.94, 95%CI (0.86, 1.02)], disease-free survival rate [RR=1.02, 95%CI (0.89, 1.17)], positive lymph node rate [RR=0.86, 95%CI (0.47, 1.57)], and seminal vesicle invasion rate [RR=1.09, 95%CI (0.74, 1.59)] between single prostatectomy and preoperative hormone therapy plus prostatectomy, while there was significant difference in positive surgical margin rate [RR=0.46, 95%CI (0.32, 0.66)]. This suggested that preoperative hormonal therapy prior to prostatectomy did not improve survival rate, positive lymph nodes rate and seminal vesicle invasion rate; however, there was a significant reduction in the positive surgical margin rate. Related Articles | Metrics

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Comparison of rabbit intraocular hypertension models prepared by three different material injections Wang Feng-yun, Lu Xiao-he, Bai Lang, Zhang Jing, Zhang Cai-xia 2010, 14 (11):  1935-1938.  doi: 10.3969/j.issn.1673-8225.2010.11.009 Abstract ( 371 )   PDF (317KB) ( 550 )   Save BACKGROUND: Currently, there are many studies concerning the pathogenesis, process, and damage of glaucoma, however, there is not an ideal glaucoma model. OBJECTIVE: To prepare rabbit intraocular hypertension models using three different material injections, and to verify the practical value of intraocular hypertension models. METHODS: Thirty New Zealand rabbits were divided randomly into 3 groups, with 10 animals in each group. One eye of each rabbit was served as the experimental eyes and the other eye as control eyes. Autoblood, methyl cellulose, C3F8 was injected into the anterior chamber of the experimental eyes, and the normal saline was injected into the control eyes. The intraocular pressure (IOP) was monitored prior to injection and at hours 0, 24, 36, 48, 72, 96, 120 and 168 after injection. RESULTS AND CONCLUSION: Intraocular hypertension models could be induced by injecting 3 kinds of materials, and the IOP was obviously increased after injection (P < 0.05), and the ranges and periods of increasing were varied. The periods of increasing of 3 materials were 1, 3 and 7 days, respectively, which could maintain for longer time for a second injection. The IOP ranged 1.86-6.65 kPa, and mild anterior segment inflammation could be found. The experiment demonstrated that intraocular hypertension models using three different material injections are ideal models, which is characterized by simple, reliable and controllable. The suitable model can be selected for acute or chronic glaucoma research. Related Articles | Metrics

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A modified method for primary culture of human glomerular mesangial cells Yin Yan-zhi, Zhang Li-mei, Ju Jian-wei, Wang Hong-mei, Wang Sha 2010, 14 (11):  1939-1942.  doi: 10.3969/j.issn.1673-8225.2010.11.010 Abstract ( 502 )   PDF (483KB) ( 846 )   Save BACKGROUND: Primary culture of glomerular mesangial cells was less achievement ratio, short survival time, and less passage times. In particular, extraction of renal glomerulus remains difficult for culturing highly pure mesangial cell. OBJECTIVE: To establish a more simple, high successful rate and good reproducibility method of human mesangial cells in primary culture. METHODS: Kidneys isolated from induction of labor with water bag voluntary were cut into pieces, and human mesangial cells were cultured with eugenic selection methods. Morphology was observed using inverted phase contrast microscope and transmission electron microscope, cell phenotype was detected using immunohistochemical method, and vimentin expression was observed using laser scanning confocal microscope. RESULTS AND CONCLUSION: The cultured mesangial cells were fusiform-shaped, irregular star-shaped, and slender. Organelle was rich in cytoplasm, cell process was clear, and microvillus was observed on the cell membrane. The cells expressed α-actin, myosin, vimentin, desmin but not expressed cytokeratin and VIII factor. Laser scanning confocal microscope demonstrated that vimentin expression was positive and had the characteristics of fiber bundles. This suggested that the cultured intercapillary cells were coincidence with the characteristics of mesangial cell. The renal cortical tissue combined eugenic selection method was a simple and efficient method to culture human mesangial cells. Related Articles | Metrics

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Genetic recombinant prokaryotic expression of augmenter of liver regeneration and preparation of polyclonal antibodies in rats Liu Zheng-fang, Wang Jian-ming, Wang Lan, Zeng Xiao-yun, Xiong Ling, Luo Zhi-xiu, Wu Jun-yi 2010, 14 (11):  1943-1947.  doi: 10.3969/j.issn.1673-8225.2010.11.011 Abstract ( 414 )   PDF (415KB) ( 552 )   Save BACKGROUND: An abroad study reported the distribution and expression of augmenter of liver regeneration (ALR) in the central nervous system. There are few literatures on how to prepare and evaluate ALR protein polyclonal antibody in recombinant rats, and how to construct prokaryotic expression vector. There are no reports concerning ALR in the central nervous system in China. OBJECTIVE: To express ALR fusion protein in E. coli BL21 and prepare and identify polyclonal antibody. METHODS: RNA was extracted from the hippocampus of Sprague Dawley rats. The prokaryotic expression plasmid pET28a-ALR was constructed and the positive recombinant plasmid was transformed into BL21. Protein ALR was expressed by inducing transformed BL21 with Isopropyl-β-D-thiogalactopyranoside (IPTG) and purified by Ni2+ affinity chromatography column after immune the rabbit for 4 times, the serum of rabbits was extracted from heart as polyclonal antibody. The titer and specificity of the rabbit’s antiserum was respectively measured by ELISA and Western blotting. The following parameters were measured: construction of prokaryotic expression plasmid pET28a-ALR; pET28a-ALR recombinant enzyme digestion evaluation; results of ELISA and Western-blotting. RESULTS AND CONCLUSION: Expecting bands were obtained by double enzyme digestion electrophoresis, respectively    5.3 kb and 0.4 kb. Nucleotide sequence analysis verified that prokaryotic expression vector pET28a-ALR was successfully constructed. The 19 ku fusion protein was successfully expressed. The titer of the antiserum measured by ELISA could achieve  1: 2 000. This indicated that antibody and purified recombinant ALR had a good reaction, and high titer, could meet the experimental require. Western blotting analysis proved that the antibody could identify the prokaryotic expression product of ALR. Prokaryotic expression system expressed ALR fusion protein, prepared and purified polyclonal antibody of ALR protein, and could meet the experimental require of ALR immunoblotting. Related Articles | Metrics

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Cloning of human csp-B matrix attachment region sequence and construction of its retrovirus vector Zan Yu-xi, Wang Li, Zhang Jun-he, Wang Tian-yun 2010, 14 (11):  1948-1950.  doi: 10.3969/j.issn.1673-8225.2010.11.012 Abstract ( 320 )   PDF (364KB) ( 519 )   Save BACKGROUND: Matrix attachment region (MAR) are DNA elements that bound to the nuclear matrices after chromatin digested with restriction endonuclease. Plenty of studies have shown that MAR considered as initial point of DNA replication or transcription of regulatory gene. Thereby, construction of MAR expression vector can elevate the overall level of transgene expression, enhance stability of exogenous gene, as well as increase frequency of stable transfectant cells. OBJECTIVE: To construction pLXSN-CAT recombinant retrovirus vector that containing chloramphenicol acetyltransferase (CAT) via cloning MAR sequence of human, and to explore the influence of MAR on the gene expression. METHODS: An open experiment was performed at the Department of Biochemistry and Molecular Biology, Xinxiang Medical College from September 2007 to December 2007. The PLXSN-CAT vector of CAT was constructed by the laboratory. TaqDNA polymerase, T4 DNA ligase, DNA Marker, restriction enzyme BamHⅠ, agarose gel DNA purification kit, as well as plasmid purification kit were purchased fromTakara Biotechnology (Dalian) Co., Ltd. The sequence of csp-B MAR was amplified by polymerase chain reaction (PCR) method applied to human DNA. The fragment was inserted into retrovirus vector PLXSN-CAT plasmid. The recombinant plasmid was verified by double digestion and DNA sequencing. RESULTS AND CONCLUSION: The length of specific fragment applied by PCR was 931 bp, and the recombinant plasmid PLXSN-CAT-MAR presented two bands: 5.9 kb and 931 bp using respective restriction enzymes BamHⅠ. The sequence of MAR was confirmed by blasting to Genbank (serial numobr: M62716). It suggested that MAR had been cloned into PLXSN-CATR vector correctly. The recombinant retrovirus vector PLXSN-MAR was successfully constructed. Related Articles | Metrics

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Construction and identification of small interfering RNA expression plasmid target to angiotensinogen Wu Li-hua, Yang Ru-de 2010, 14 (11):  1951-1954.  doi: 10.3969/j.issn.1673-8225.2010.11.013 Abstract ( 386 )   PDF (483KB) ( 520 )   Save BACKGROUND: In mammalian cells, introduction of double-stranded small interfering RNA (19-25 bp) can cleave and destroy the cognate RNA, which can result in suppression of gene expression. OBJECTIVE: To construct siRNA expression plasmid for interference angiotensinogen (AGT), thereby, to resist AGT expression in adipose cells. METHODS: The mRNA sequence of AGT gene was searched from NCBI (NM000029). Utilize of GenScript siRNA technology, AGT-siRNA oligonucletides were chemically synthesized and inserted into pRNAT-U6.1/Neo vector after annealing, then transformed into TOP10. The recombinant plasmid was identified by restriction endonuclease and DNA sequencing. RESULTS AND CONCLUSION: The recombinant plasmid psiRNAT-U6.1/Neo-AGT was obtained by connecting 19 bp segment containing AGT-mRNA sequence to pRNAT-U6.1/Neo. After EcoR Ⅰ and Hind Ⅲ digestion, 351 bp segment was obtained from empty vector, and 397 bp fragment band was obtained form recombinant plasmid, which was coincidence to the expectation. DNA sequencing showed Targeting siRNA oligonucleotides were correctly inserted into the eukaryotic expression vector pRNAT-U6.1/Neo without base mutation. The interference vector psiRNAT-U6.1/Neo-AGT was successfully constructed. Related Articles | Metrics

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Comparison of rabbit degenerative intervertebral disc models constructed by anulus puncture and annulus incision Luo Ping, Liu Yu-li, Chen Zhong, Lin Li-jun, Xu Yong, Huang Xiao-jun, Zhou Chu-song 2010, 14 (11):  1955-1958.  doi: 10.3969/j.issn.1673-8225.2010.11.014 Abstract ( 271 )   PDF (292KB) ( 548 )   Save BACKGROUND: Constructing animal model of intervertebral disc degeneration which more faithfully mimics the pathologic hallmarks of human intervertebral disc degeneration can be a beneficial assistance for further intervertebral disc degeneration therapy. However, there is not an accepted optimal model for intervertebral disc degeneration study. OBJECTIVE: To compare the rabbit model of degenerative intervertebral disc constructed by anulus puncture and anulus incision. METHODS: Totally 32 New Zealand white rabbits were randomly divided into the anulus puncture group and annulus incision group. Intervertebral disc of L3-6 was exposed by extro-peritoneal approach, and the discs were injured by puncturing the anulus or cutting the anulus. The deep and direction were controlled. Pathological change of intervertebral disc was checked with MRI and histopathological examination at weeks 2, 4, 12, and 20 after operation. RESULTS AND CONCLUSION: At week 4 after operation, the area of nucleus gelatinosus was deflated with enlarged anulus fibrosus, T2-weighted image (T2WI) declined, blurred, and the height of intervertebral space was also decreased, the grade of T2 value in the anulus puncture group was lower than that of the annulus incision group (P < 0.05); with time prolonged, T2 scores increased, and the intervertebral space narrowed, which reached a peak at week 20 after operation. The differences had no significance. The histological sections demonstrated that the cell content in nucleus pulposus was increased gradually. The rabbit model of intervertebral disc degeneration can be successfully constructed by the methods of anulus puncture and annulus incision. The degeneration of incision model is more severe than that of puncture model. Anulus puncture method can faithfully mimic intervertebral disc degeneration after damage in human being. Related Articles | Metrics

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Establishment and evaluation of an animal model of stress urinary incontinence Wang Hao-yu, Du Xiao-wen, Xu Jian-wei, Zhu Yong-feng, Wu Hui-ling, Hu Jun-biao, Jin Fan, Lü Rui-ping 2010, 14 (11):  1959-1962.  doi: 10.3969/j.issn.1673-8225.2010.11.015 Abstract ( 507 )   PDF (436KB) ( 997 )   Save BACKGROUND: Although drug treatment, physics-behavior therapy, and postoperative therapy have been commonly used to treat stress urinary incontinence (SUI), there is still no satisfactory treatment at present. OBJECTIVE: To build a stable animal model simulating stress urinary incontinence (SUI) by bilateral transaction of pudendal nerve and nerves innervating pelvic floor muscles, including iliococcygeous muscle and pubococcygeous muscle. METHODS: A total of 18 6-week-old female SD rats weighing (199.44±8.41) g were randomly divided into 3 groups: normal, model, and sham-surgery groups, with 6 rats in each group. Rats in the model group underwent bilateral transaction of pudendal nerves and nerves innervating iliococcygeous/pubococcygeous muscles, while rats in the sham-surgery group had same procedures except nerve transaction. The normal group did not undergo any operation. Each rat was subjected to measure leak point pressure (LPP) at 2 weeks after the operation. After the measurement of LPP, cross sections of connection area of bladder and urethra were sent to histology. RESULTS AND CONCLUSION: One rat in the sham-surgery group died at 1 week after the operation. The LPP of model group decreased significantly by approximately 33% compared with the normal group (P < 0.05); however, there was no significant difference in LPP between sham-surgery and normal groups (P > 0.05). The results of histology showed loosely arrangement and atrophy of urethral striated muscle fibers in rats of the model group. Bilateral transaction of pudendal nerves and nerves innervating to iliococcygeous/ pubococcygeous muscles resulted in SUI in rats stably. Related Articles | Metrics

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Changes of glucocorticoid-induced osteonecrosis of the femoral head under mechanics load Wang Zhi-gang, Ma Xin-long 2010, 14 (11):  1963-1966.  doi: 10.3969/j.issn.1673-8225.2010.10.016 Abstract ( 373 )   PDF (430KB) ( 457 )   Save BACKGROUND: Femoral head necrosis induced by glucocorticoid is typical under mechanics load. OBJECTIVE: To create the rat models of the femoral head necrosis by long-term glucocorticoid treatment, and to observe the changes of femoral head necrosis induced by glucocorticoid under mechanics load. METHODS: Wistar rats with 4-month old were randomly divided into the experimental and control groups. After the buttocks injection of dexamethasone 20mg/kg weakly for 8 weeks, rats in the experimental group were made to run on the running machine at a speed of 1 km/h to forme the hyper-weight model; at the same time, those in the control group were with the normal physiological weight. Rats in both groups were sacrificed at weeks 2, 4, 6 and 8. The left legs were taken for the biomechanics test on the EnduraTEC ELF3200, the biggest vertical deformation and stiffness were calculated, the number of empty osteocyte lacunas was calculated, and the cumulative absorbance value was compared between two groups.  RESULTS AND CONCLUSION: The femoral head presented with necrosis at 8 weeks in the experimental group, and the width of bone trabecula, and the stiffness of which were smaller than that of the control group, but the biggest vertical deformation was greater the concurrent control group (P < 0.05). The Bcl-2 cumulative absorbance value of 2 groups had significant differences at 4 weeks (P < 0.05), which was increased with time prolonged in the experimental group (P < 0.05). The results demonstrated that: in glucocorticoid-induced osteonecrosis of the femoral head, mechanics load induces the obviously osteocyte necrosis, more cartilage repair, and trabeculae rupture, which is considered as the direct reason of the necrosis of the femoral head. The typical femoral head necrosis is presented Related Articles | Metrics

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Effects of fluorid on biomechanical parameters of the femur in young rats Wu Tian-xiu, Liao Jin-min, Chen Yan, Huang Lian-fang, Chen Wen-shuang 2010, 14 (11):  1967-1970.  doi: 10.3969/j.issn.1673-8225.2010.11.017 Abstract ( 297 )   PDF (367KB) ( 521 )   Save BACKGROUND: Fluoride treatment of osteoporosis has been controversial. Literatures addressing the effect of fluoride on bone bio-mechanical parameters of femur in young rats are few. OBJECTIVE: To study the effects of fluoride on bone biomechanical parameters of femur in young rats. METHODS: Ninety 2-month-old SPF Sprague Dawley rats, half male and female, were randomly divided into 9 groups: control group (young, adult and long-time) and drug-administered group (young high-fluoride, young low-fluoride, adult high-fluoride, adult low-fluoride, long-term high-fluoride and long-term low-fluoride). Rats in the control group were orally administered with physiological saline, while in the drug-administered group were given orally with different dose fluoride at the corresponding times. After experiment, rats were sacrificed under anaesthesia. Three-point bending test was performed at the left femur. The effects of fluoride on maximum load and rigidity of femur were measured. RESULTS AND CONCLUSION: Compared with young control group, the maximum load and the rigidity of femur in the young high-fluoride group were decreased by 13.18% and 13.61%, respectively (P < 0.05), which had no dramatically difference in the young low-fluoride group. Compared with long-term high-fluoride group, the maximum load and the rigidity of femur in the young high-fluoride were decreased by 17.22% and 17.17% (P < 0.05), which were obvious increased in the long-term low-fluoride group by 18.33% and 19.15%, respectively (P < 0.05). The maximum load and the rigidity of femur were strengthened in the adult high-fluoride and adult low-fluoride groups (P < 0.05). The results suggested that young rats are more sensitive to high-dose fluoride, which can reduce bone quality in rats. The negative effects on bone quality of rats were gradually displayed as the prolongation of the period of fluoride. Related Articles | Metrics

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Biomechanical response to stress stimulation with different intensities during early repair period of acute rotator cuff injury Wang Peng-cheng, Jin An-min, Fu Guo-jian, Li Sen, Zhang Fei, Xu Yong 2010, 14 (11):  1971-1974.  doi: 10.3969/j.issn.1673-8225.2010.11.018 Abstract ( 321 )   PDF (446KB) ( 586 )   Save BACKGROUND: Continuing stress stimulation has been confirmed to promote repair of rotator cuff injury. This experiment attempts to further quantify the intensity of stress stimulation in order to reach a better therapeutic effect in the entire process of rotator cuff injury repair. OBJECTIVE: To explore the biomechanical performance of stress stimulation with different intensities on repair of acute rotator cuff injury. METHODS: Sixteen male New Zealand rabbits were randomly divided into high intensity, low intensity and control groups, with 12 animals in each group. Rabbits were received a rotator cuff acute injury and reconstructing insertion of supraspinatus tendon on greater tuberosity of humerus. At 1 week after operation, rabbits were treated by continuous passive motion (CPM) apparatus, which was -60° to 60° flexion-extension for the high intensity group, and -15° to 15° for the low intensity group. At 2 weeks after operation, the CPM was performed every other day, with 10 minutes per time; which was changed to daily training from week 3 with the same frequently, and 15 minutes per time at week 4. Rabbits in the control group were housed freely. Each 4 animals were sacrificed at weeks 2, 3 and 4 after operation, and the largest tensile strength and severity at osteotendinous junction were compared.  RESULTS AND CONCLUSION: The maximum tensile intensity and stiffness of the supraspinatus tendon in the low intensity group were greater than that of the control and high intensity group (P < 0.05), which was smallest in the high intensity group. All the breakage point existed at the intra-articular portion. The result demonstrated that low intensity stimulation exhibits superior repair effect in acute rotator cuff injury, while high intensity stimulation is harmful to the early repair of acute rotator cuff injury. Related Articles | Metrics

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Isokinetic test for flexor and extensor muscle strength of hip joint: Characteristics of students in physical education colleges Jia Fu-chi, Hu Xiao-yan, Cui Ba-ter, Chen Yu-juan, Li Li 2010, 14 (11):  1975-1978.  doi: 10.3969/j.issn.1673-8225.2010.11.019 Abstract ( 291 )   PDF (310KB) ( 511 )   Save BACKGROUND: Isokinetic test system has been widely used to evaluate the muscle function levels. Whereas, the reports concerning effect of sport training on the development of muscle strength of hip joint in college students using isokinetic testing are few. OBJECTIVE: To assess the extensor and flexor muscle strength of hip joints in college students with isokinetic testing system. METHODS: Totally 40 volunteers, including 20 male and 20 female students were selected. Kinitech isokinetic testing system, produced by Australian Kylingk Company, was used to test the flexion and extension muscle of hip joints as the sequence of centripetal followed by centrifugal. The test speed was 60 (°)/s as slow speed, 180 (°)/s as middle speed, 240 (°)/s as high speed. Peak torque (PT) and relative values of extension and flexion muscles, total power (TP) and relative total power were measured. RESULTS AND CONCLUSION: At the respective speed, PT and relative values of female students were greater than that of female students (P < 0.01). At the speed of 60, 120 and 240 (°)/s, flexor PT was smaller than that of extensor PT in male students, which was opposite in the female students, but the difference had no significance (P > 0.05). At the respective speed, TP of males were greater than that of female students (P < 0.01). In male students, TP and relative values of flexor were smaller than extensor at speed of 60 (°)/s and 120 (°)/s (P < 0.01), and had difference at speed of 240 (°)/s (P < 0.05). In female students, TP and relative values had significant difference between flexor and extensor at speed of 60 (°)/s (P < 0.01) and at speed of 120 (°)/s and 240 (°)/s (P < 0.05). At the respective speed, PT and relative values, TP and relative values in male students are all greater than females. These values decrease with the speed increasing. At the respective values, values of male students are bigger than females. Related Articles | Metrics

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Creep mechanical properties of middle cerebral artery in rat cerebral atherosclerosis models Xu Dong-hui, Li Xin-ying 2010, 14 (11):  1979-1982.  doi: 10.3969/j.issn.1673-8225.2010.14.020 Abstract ( 371 )   PDF (408KB) ( 503 )   Save BACKGROUND: Creep is a visco-elastic biological material form of solid performance. It is necessary to understand the creep mechanical properties of middle cerebral artery in prevention and treatment of cerebral arteriosclerosis and cerebral hemorrhage. OBJECTIVE: To compare the creep rheological properties of middle cerebral artery between the normal and atherosclerotic animal model, to identify the effects of atherosclerosis on the creep properties of middle cerebral artery. METHODS: Male SD rats were randomly divided into 2 groups, in the model group, rats were injected vitamin D3 + nicotine + high-fat diet fed to prepare atherosclerosis models; in the control group, rats were fed with normal diet. Rats were killed at 4 weeks and the middle cerebral artery specimens were obtained. The creep experiments were performed with electron Shimadzu universal testing machine. The stress rate was 0.01 MPa/s, with 7 200 s experimental time. A total of 100 numbers were collected and analyzed using one linear regression analysis. The creep curve, as well as the variation of strain and time was recorded. RESULTS AND CONCLUSION: The middle cerebral artery creep changed faster at the initial 600 s in the normal and atherosclerotic rats, and then slowly decreased. The creep of normal rats were greater than that of the atherosclerotic artery specimens at the 7 200 s (50.38%, 48.26%, P < 0.05). The results demonstrated that the middle cerebral artery have different mechanical properties of creep in normal and atherosclerotic rats. The creep curve is changed in an exponential relationship, and the atherosclerosis has certain effect on the creep characteristics of middle cerebral artery. Related Articles | Metrics

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Effect of sodium hyaluronateon on vascular endothelial growth factor mRNA expression of the synovium in rabbits with osteoarthritis Zhou Jian-lin, Liu Shi-qing, Qiu Bo 2010, 14 (11):  1983-1986.  doi: 10.3969/j.issn.1673-8225.2010.11.021 Abstract ( 375 )   PDF (423KB) ( 462 )   Save BACKGROUND: Pathogenesis of osteoarthritis is poorly understood, however, studies have demonstrated that vascular endothelial growth factor (VEGF) is involved in the progression of osteoarthritis. OBJECTIVE: To detect the changes of VEGF mRNA expression of synovium in rabbit osteoarthritis and to evaluate the effect of sodium hyaluronate on its expression. METHODS: Twenty four white rabbits were divided into the normal control, physiologic saline, and sodium hyaluronate groups. The unilateral anterior cruciate ligament transection (ACLT) was performed in the physiologic saline and sodium hyaluronate groups. At weeks 4 after operation, rabbits in the physiologic saline group were injected 0.3 mL physiologic saline, and in the sodium hyaluronate group received 10 g/L sodium hyaluronate injection, once per week for 5 successive weeks. All the animals were sacrificed at week 10 after operation. The cartilage changes on the medial femoral condyles were graded separately. VEGF expression of synovium was detected by using real time polymerase chain reaction (RT-PCR). RESULTS AND CONCLUSION: The macroscopic score showed that the cartilage degeneration in the physiologic saline group was significantly more severe than that of the normal control and sodium hyaluronate groups (P < 0.05). The expression of VEGF mRNA was obviously decreased in the physiologic saline group than that of the normal control group (P < 0.05), of which was increased in thesodium hyaluronate group, but still smaller than the normal control group (P < 0.05). The results demonstrated that the decreased VEGF expression in synovium may involved in the progression of osteoarthritis, and sodium hyaluronate has protective effect on articular cartilage by up-regulating the VEGF expression. Related Articles | Metrics

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Influence of beta nerve growth factor in vibrissa follicle bulge region on the burn wound healing of neonatal rats Zeng Jun, Chen Deng-bang, Dai Lü-xia, Lin Sen 2010, 14 (11):  1987-1991.  doi: 10.3969/j.issn.1673-8225.2010.11.022 Abstract ( 367 )   PDF (546KB) ( 734 )   Save BACKGROUND: The surroundings stimulation, such as wound and chemical injury, will result in changes of hair follicle stem cells (HFSC). Up to date, few articles indicate the effect of burn on HFSCs.  OBJECTIVE: To investigate the changes of β-nerve growth factor (β-NGF) level in vibrissa follicle bulge region and its effect on the healing of neonatal rats. METHODS: A total of 30 neonatal rats were selected to prepare burn models by giving 90 ℃ boiled tap water for 3 seconds, and vibrissa follicle bulge was separated at 12, 24, 36, 48 and 60 hours postoperatively. The remained 6 neonatal rats were served as controls. Total protein concentration was measured by Brandford method. The expression of β-NGF was detected by ELISA. Meanwhile, the tissues in each time points were collected for frozen section and immunohistochemistry. RESULTS AND CONCLUSION: Among the tested total protein, the expression of the β-NGF increased after 36 hours, reached a peak at 60 hours after burn. The identical changes were found in tissue slices. The image analysis demonstrated that the β-NGF protein detection was coincident with β-NGF expression in tissues. During 12 hours to 60 hours after burn, the expression of the β-NGF in the bulge area peaked at 60 hours; simultaneously, the repairing effect of healing also reached a peak. Bulge area is a nest of HFSC, the enhanced expression of β-NGF may have some positive significance to the differentiation of the HFSC and tissue repair. Related Articles | Metrics

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May HOXD13-zinc finger protein 548 complex develop in foot tissue during human embryonic development? Wang Li-li, Xin Na, Sun Kai-lai 2010, 14 (11):  1992-1994.  doi: 10.3969/j.issn.1673-8225.2010.11.023 Abstract ( 435 )   PDF (321KB) ( 519 )   Save BACKGROUND: The HOX gene family is a highly conserved transcription factor family, which affects the formation of basic axis and secondary axis during embryonic development, at the same time, it plays a pivotal role in the development of the central nervous system, axial skeleton, stomach intestine, urogenital system and external genitalia. OBJECTIVE: To investigate the interactional protein of HOXD13 in human embryo foot development at 14 weeks pregnancy. METHODS: Fetal foot tissues were harvested and protein extraction was performed using protein extraction kit, 6 μg HOXD13 antibody was added for immunoprecipitation experiment, which aims to obtain HOXD13 interactional proteins, while the non-specific IgG served as a negative control. Subsequent to Bradford staining, the sediment protein bands were cut for mass spectrometry. RESULTS AND CONCLUSION: We obtained an interactional protein of HOXD13 in the human embryo foot at 14 weeks pregnancy, at the relative molecular mass of approximately 60 000. The protein was identified as zinc finger protein 548 using mass spectrum. HOXD13 may play a very important role in transcriptional control by HOXD13-zinc finger protein 548 protein complex in the human embryo foot at 14 weeks pregnancy. Related Articles | Metrics

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Effects of heparinase Ⅰ on syndcan-1 and extracelluar signal regulated kinase expression in human umbilical vein endothelial cells following hypoxia/reoxygenation Luo Hai-yan, Hu Chang-lin 2010, 14 (11):  1995-1999.  doi: 10.3969/j.issn.1673-8225.2010.11.024 Abstract ( 433 )   PDF (595KB) ( 501 )   Save BACKGROUND: Heparinase can induce syndecan-1 shedding from tumor cells, and macrophage motion may correlate with biosynthesis regulation of heparan sulfate proteoglycan. OBJECTIVE: To investigate the effects of heparinaseⅠon syndecan-1 and extracelluar signal regulated kinase 2 (ERK2) in human umbilical vein endothelial cells (HUVECS) with hypoxia/reoxygenation injury. METHODS: heparinaseⅠ-precultured HUVECS were treated with hypoxia/reoxygenation. Immunohistochemistry staining, RT-PCR and western blot were applied to detect HUVECS syndecan-1 and ERK2 expression. RESULTS AND CONCLUSION: Expression of syndecan-1 and ERK2 was increased in HUVECS following hypoxia/reoxygenation treatment. HeparinaseⅠ significantly upregulated the expression of syndecan-1 and ERK2 in HUVECS with hypoxia/reoxygenation treatment. Syndecan-1 and ERK2 expression was positively related. Results show that syndecan-1 and ERK2 participate the pathophysiology of HUVECs hypoxia/reoxygenation injury. HeparinaseⅠ influences ERK2 expression by regulating syndecan-1. Related Articles | Metrics

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Effect of autumn and winter seasons on expression of surfactant associated protein A and interleukin-6 in healthy rat lung tissues Lu Zhi-gang, Wang Ya-li, Zhang Ming-quan, Guo Jin-qiao, Shi Xu-liang, Gao Yang 2010, 14 (11):  2000-2003.  doi: 10.3969/j.issn.1673-8225.2010.11.025 Abstract ( 426 )   PDF (442KB) ( 519 )   Save BACKGROUND: Previous study has confirmed that seasonal variation and surfactant associated protein A (SP-A) and interleukin-6 (IL-6) exhibit a significant role in lung immune and defense function. However, the effect mechanism of them remains unclear. OBJECTIVE: By observing the changes of SP-A and IL-6 of male rats in autumn and winter, to explore the effects of seasonal changes on the nonspecific immunity of lung of normal rats, and to provide new ideas for the experimental basis to the cognition of pathology and physiology mechanism for seasonal attack of respiratory system diseases. METHODS: Male SD rats were purchased from each seasons and divided into groups in chronological order. All rats were housed with normal forage and drank freely under room temperature. Then the rats were sacrificed by decapitation before 18 o’clock at corresponding solar term, and the lung tissues were analyzed by RT-PCR. RESULTS AND CONCLUSION: Compared to the other groups, the expression of SP-A and IL-6 in the Winter Begins group were smaller, which demonstrated that immune function of lung was existed the seasonal rhythm of lower in winter and higher in autumn. The SP-A and IL-6 are main material basis of immune function. Related Articles | Metrics

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Influence of different Chinese medical compounds on mouse exercise capacity and biochemical indexes Wang Yan-qiong, Qu Hong-lin, Yang Yong-liang 2010, 14 (11):  2004-2007.  doi: 10.3969/j.issn.1673-8225.2010.11.026 Abstract ( 313 )   PDF (324KB) ( 414 )   Save BACKGROUND: Many studies concerning influence of traditional Chinese medicine on animal movement capacity have been conducted. However, studies on the improvement of exercise capacity by traditional Chinese medicine are few, especially comparisons among traditional Chinese medicine OBJECTIVE: To explore the influence of different traditional Chinese medicine preparations on the exercise capacity in mice to screen compound preparations with precise effects. METHOD: A total of 80 8-week-old pure male Kunming mice were randomly assigned to 8 groups (n=10): control and compound 1-7 groups. There were no significant differences in body mass of all groups. The compound 1 comprised Xuejie and Shexiang; compound 2 comprised Qingui, Baizhu and Baishao; compound 3 comprised Baizhu, Baizhi and Sanqi; compound 4 comprised Baizhi, Ruxiang, Danggui and Radix glycryrrhizae; compound 5 comprised Guiwei; compound 6 comprised Sichuan Honghua and Danshen; compound 7 comprised Danggui and Shexiang. Of the 7 prescriptions, compound 1-5 were empirical formula, compound 6 was common medicine used during training, and compound 7 was prescription of traditional Chinese medicine. The 7 compound groups were respectively treated with 1.17, 0.78, 0.78, 1.17, 7.02, 5.53, and 11.70 g/kg per day for 4 weeks, and control group was intragastrically infused with normal saline. From day 4 to 6, load-free swimming training was performed 20 minutes a day for 3 days, followed by swimming for 6 days a week after 1 day of rest. The intensity was gradually increased from 30 minutes per day by 10 minutes per day for 2 hours, followed by loading swimming with load of 2% of the body mass, increased by 1% per day until the final experiment. During the entire procedure, a great intensity swimming training was performed every week to make the mice exhausted. Mice training and serum creatine kinase and lactate dehydrogenase 4 weeks following treatment were observed. RESULTS AND CONCLUSION: A total of 80 mice were included in the final analysis. The compounds showed little influence on mice body mass; the compound 2 and compound 6 groups had longest exhaustive time. All compounds inhibited serum creatine kinase activity following exercise, in particular compound 6 and 2 (P < 0.05). Compared with control group, the serum lactate dehydrogenase activity of each compound group was decreased, in particular, compound 2 and 6 groups showed significant differences (P < 0.05). Compound 2 and 6 significantly enhanced the exercise capacity of mice and delayed injury of body. The mechanism may highly relate to components in the compounds, Danggui, Sichuan Honghua and Danshen. Related Articles | Metrics

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Repair of articular cartilage injury using tissue-engineered cell transplantation Zhao Zhi-jun, Li Shi-he 2010, 14 (11):  2010-2013.  doi: 10.3969/j.issn.1673-8225.2010.11.028 Abstract ( 314 )   PDF (391KB) ( 479 )   Save BACKGROUND: Simple medication therapy can not promote the healing of articular cartilage defects effectively; the insufficient sources of autogenous cartilage also limited cartilage transplantation.  OBJECTIVE: The types of articular cartilage injury and local micro-environment changes were analyzed to summarize the progress of tissue engineered seed cell transplantation and cell transplantation therapy for repairing articular cartilage injury. METHODS: The databases of PubMed database (http://www.ncbi.nlm.nih.gov/sites/entrez/) and CNKI (http://www.cnki.net/) were retrieved with the key words of “Tissue engineering, cell transplantation, articular cartilage defects”, and the literature was limited to English and Chinese languages. Studies concerning articular cartilage injury or was closely related to tissue engineered seed cells transplantation were included. Repetitive studies were excluded. Survival and migration of seed cells, as well as the recovery of joint function and adverse reactions were served as evaluation index. RESULTS AND CONCLUSION: A total of 201 literatures were seized by computers, according to the inclusion criteria, papers concerning tissue engineering cells for repairing articular cartilage injury were analyzed. Clinically, articular cartilage damage was commonly resulted by traumatic arthritis, which is difficult to repair due to the poor self-repair capacity of articular cartilage. The emergence of tissue engineered cell transplantation brought new hope for articular cartilage injury healing. Cell transplantation has obtained good effects in articular cartilage repair, but the technology is still room for improvement. How to technically improve the tissue-engineered 3 elements, namely, cells, scaffold materials, and biological activity factors were the research focuses. Autologous cartilage regeneration was the theoretical support for articular cartilage defects repairing, which needs to optimize its regeneration and to maintain a more stable chondrocyte phenotype in further research. The tissue-engineered cells had received good effects in repairing articular cartilage injury, and it is expected to be a new clinical treatment for articular cartilage injury with the in-depth study of tissue engineering. Related Articles | Metrics

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Notch signaling pathway, articular cartilage development and osteoarthritis Liu Xian-wen, Zhu Song-song, Hu Jing 2010, 14 (11):  2014-2017.  doi: 10.3969/j.issn.1673-8225.2010.11.029 Abstract ( 466 )   PDF (396KB) ( 777 )   Save BACKGROUND: Osteoarthritis, often accompanied by cartilage abnormal signal transduction, suggesting that intracellular signal transduction may play an important role in osteoarthritis genesis and development. OBJECTIVE: To review recent advances in the research of Notch signaling pathway, and explore the mechanisms of chondrogenic differentiation of bone marrow mesenchymal stem cells and its role in cartilage development and chondrogenesis. To analysis the role of dysregulated Notch signaling in the development of osteoarthritis. METHODS: Pubmed was undertaken to identify the relevant articles published from March 1919 to February 2009 with the key words of “Notch signaling pathway, osteoarthritis, chondrocytes, chondrogenesis, bone marrow stem cells” in English. Simultaneously, CNKI Database was searched for relevant articles published from February 2004 to September 2008 with the key words of “Notch signaling pathway, osteoarthritis, chondrocytes, chondrogenesis, bone marrow stem cells” in Chinese. Articles with the Notch signaling pathway-related research, Notch signaling pathway of bone marrow-derived mesenchymal stem cells, differentiation of cartilage relative to the mechanism studies, and the Notch signaling pathway disorders and osteoarthritis research were included. Reproducibility articles were excluded. Development regulation of articular cartilage and cartilage cell proliferation were considered as evaluation indexes. RESULTS AND CONCLUSION: Of 632 articles retrieved, 30 were included in our study according to the inclusive and exclusive criteria. The Notch pathway is a highly conserved signaling mechanism involved in many processes determining cell fate during development and maintenance of homeostasis of mature tissues in the adult organism. Activation of Notch occurs following Notch receptor/ligand engagement upon cell-cell contact, which initiates translocation of the Notch intracellular domain to the nucleus and activation of target genes. It has been demonstrated that the Notch signaling plays an important role in regulating articular cartilage development and chondrocyte proliferation and differentiation. In recent studies abnormality of notch signaling was also found in disease, which suggests that notch signaling is involved in osteoarthritis initiation and development. Further study of the specific mechanism of notch signaling in the initiation and development of osteoarthritis will enable us to develop targeted approaches in the treatment of osteoarthritis, thus bring more strategies in the future treatment of osteoarthritis. Related Articles | Metrics

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Construction and application of tissue-engineered adipose tissue Li Dong-jun, Jia Quan-zhang 2010, 14 (11):  2018-2021.  doi: 10.3969/j.issn.1673-8225.2010.11.030 Abstract ( 448 )   PDF (469KB) ( 465 )   Save BACKGROUND: Soft-tissue defect in plastic surgery is a major problem, and currently all of methods used for clinical treatment have certain shortcomings. OBJECTIVE: To investigate the research status of adipose tissue engineering, to analyze the current deficiencies in the application of the program, and to outlook its possible future in clinical application. METHODS: A computer-based online search was conducted in Medline and CNKI with the key words of “adipose tissue engineering, mesenchymal stem cells, derived from adipose tissue” in both Chinese and English from January 1991 to May 2009. The related articles about fat tissue and tissue-engineered building were included, but the duplicated studies were excluded. RESULTS AND CONCLUSION: A total of 342 documents were retrieved firstly. According to the inclusion criteria, 33 articles were included in the final analysis. During the exploration of treatment for soft-tissue defect, tissue engineering technology provided a new method by combining the seed cells amplifying in vitro with biomaterial to repair some tissue or organ defects, reconstruct physiological function, and finally to achieve the ultimate goal of non-damage repairment of organic defect and reconstruction of morphology, structure and function. Nowadays there are several successful strategies of constructing adipose tissue in treatment for soft tissue defect, which will fill in the blanks in the repairment of adipose tissue defect. Related Articles | Metrics

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Peroxisome proliferator-activated receptor-gamma coactivator 1 and skeletal muscle adaptation Deng Lei, Wang Song, Niu Jie2 2010, 14 (11):  2022-2025.  doi: 10.3969/j.issn.1673-8225.2010.11.031 Abstract ( 424 )   PDF (439KB) ( 567 )   Save BACKGROUND: An increase in activity of muscle contraction can induce transcription of a variety of signaling molecules to activate a large number of gene expression within the nucleus through proprietary signaling pathway. OBJECTIVE: To review studies related the peroxisome proliferator-activated receptor-gamma coactivator 1 (PGC-1) and exercise induced skeletal muscle adaptations. METHODS: A computer-based online search of Pubmed was performed for relevant English articles published from January 1995 to January 2009 with the keywords of “PGC1, skeletal muscle, exercise, adaptations”. The relevant articles about PGC-1 and exercise induced skeletal muscle adaptations were included, and repetitive contents were excluded. PGC1 and mitochondrial oxidative metabolism, and exercise-induced skeletal muscle adaptation and PGC1s changed served as the evaluation indicators. RESULTS AND CONCLUSION: Of 57 collected articles, 34 were classified and sorted according to the criteria. Endurance training results in profound adaptations for skeletal muscle, including mitochondrial biogenesis, capillary density, and fiber composition. Transcription factors are highly dependent on coactivator molecules to regulate training physiologic adaptation processes. The majority of transcription factors target genes are involved in mitochondrial biogenesis and metabolism. These transcriptional patterns may provide a basic framework for understanding the integration of mitochondrial biogenesis and function with signaling events that exercise induced energetic properties. Post-transcription of PGC-1 alpha protein, there are a variety of protein modifications, with a variety of biological processes closely related to the possible adaptive mechanism of exercise-induced skeletal muscle. Related Articles | Metrics

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Prevention and treatment of denervated skeletal muscle amyotrophy Zhao Lei, Yan Zhi-qiang, Lü Guang-ming 2010, 14 (11):  2026-2029.  doi: 10.3969/j.issn.1673-8225.2010.11.032 Abstract ( 361 )   PDF (509KB) ( 461 )   Save BACKGROUND: Along with the medicine development, the prevention and treatment of the atrophy of denervated skeletal muscle have made a remarkable progress, but the clinical curative effect is still not satisfied. OBJECTIVE: To summarize the current status of the prevention and treatment methods of denervated skeletal muscle atrophy, and to find a more effective method for the prevention and treatment of the denervated of skeletal muscle atrophy. METHODS: Using “denervation, muscle atrophy, treatment” in English for the search term, Medline database from January 1998 to January 2008 was retrieved. Using “denervation, muscle atrophy, treatment: in Chinese for the search term, the Chinese Journal Full-text Database from January 1998 to January 2008, Wanfang Database from January 1998 to January 2008, and Journal of Clinical Rehabilitation and Tissue Engineering Research from January 1998 to January 2008 were searched. Literature was limited to English and Chinese languages. The muscular endurance and contractile force, muscle wet weight of denervated nerve, and skeletal muscle repair served as the evaluation index. The studies on the micro-surgical procedures, physical therapy, biological and chemical therapies, gene therapy for the atrophy of denervated skeletal muscle were included. Except the above methods, other therapies to the denervated skeletal muscle atrophy were excluded. RESULTS AND CONCLUSION: After peripheral nerve damage, the skeletal muscle is denervated and then inevitably atrophy occurs. Therefore, the exploration of prevention and treatment of denervated skeletal muscle atrophy has attracted the interests from many domestic and foreign scholars, will certainly become the important task and the research hot spot in the 21st century in the peripheral nerve domain. The microsurgery surgery, physiotherapy, biology and chemotherapeutics, gene therapy are the main effective prevention methods for the atrophy of denervated skeletal muscle. At present, this domain's research already presents the multiple perspectives and various tendencies. The prevention and treatment of denervated skeletal muscle atrophy has already given the pointed management, but in the improvement microcycle, but much works need to carry on in fields of improving microcirculation, preventing cell apoptosis, suppressing the collagen excessive growth, as well as adjusting myogenic regulatory factors expression using gene therapy method in the gene level, but also had the. Along with the tissue engineering, cell culture science, molecular biology, genetic engineering and so on, an unceasing development will be obtained regarding the prevention and treatment of denervated skeletal muscle atrophy. Related Articles | Metrics

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Muscle exercise and apoptosis in the skeletal muscle Zhao Shi-fei 2010, 14 (11):  2030-2034.  doi: 10.3969/j.issn.1673-8225.2010.11.033 Abstract ( 373 )   PDF (523KB) ( 682 )   Save BACKGROUND: Studies have shown that cell apoptosis can increase free radical, elevate Ca2+ concentration, and decrease mitochondrial membrane potential, ultimately reducing motor function. Therefore, understanding of relationship between apoptosis and exercise training is very important. OBJECTIVE: To summarize questions recording muscle exercise and apoptosis in skeletal muscle. METHODS: A computer-based online search of CNKI (http://dlib.cnki.net/kns50/index.aspx), and PubMed (http://www.ncbi.nlm.nih.gov/pubmed/) was performed to collect articles published between January 1990 and June 2009 with the key words “muscle exercise, apoptosis in the skeletal muscle” in Chinese and English, respectively. Articles highly correlated with current status and development of muscle exercise and apoptosis in the skeletal muscle were included: ①studies on apoptosis in the skeletal muscle; ②studies concerning exercise and apoptosis in the skeletal muscle; ③studies involving gene regulation of exercise-induced apoptosis in the skeletal muscle; ④studies on molecular mechanism of apoptosis in the skeletal muscle; ⑤studies published in latest journal or similar field. Repetitive studies were excluded. RESULTS AND CONCLUSION: After exercise, apoptosis occurs in normal muscle or muscle under pathologic state. The apoptosis of skeletal muscle cells is similar to common cell apoptosis, including karyopyknosis, plasma membrane vacuole, cell organ shrinking, and apoptotic body formation. The apoptosis process is classified into three stages: initial, effector, and degradation stages. The increased apoptosis in the skeletal muscle is the primary reason for sports fatigue. Currently, studies of gene regulation of apoptosis in the skeletal muscle focus on apoptosis regulatory factor Bcl-2 protein, tumor necrosis α and death protease cysteine aspartase. The bcl-2 inhibits opening of mitochondrial permeability transition pore, suppresses proapoptosis protein release from mitochondria, and prevents mitochondrial membrane lipid peroxidation and mitochondrial matrix Ca2+ release to play anti-apoptotic effect. Tumor necrosis family plays an important role in initializing death factor and its receptor pathways, which depends on death ligand binding death receptor to activate cysteine aspartase and lead to cell apoptosis. The exploration of the relationship between exercise intensity and apoptosis and necrosis in skeletal muscle assists better understanding of the movement mechanism of exercise-induced fatigue and effectively reducing fatigue. Related Articles | Metrics

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In vivo and in vitro animal models of intervertebral disc degeneration and repair Peng Jun, Xu Jian-guang 2010, 14 (11):  2035-2038.  doi: 10.3969/j.issn.1673-8225.2010.11.034 Abstract ( 333 )   PDF (403KB) ( 475 )   Save BACKGROUND: Animal models can be used to study specific scientific problems of intervertebral disc biology. Model of disc degeneration is mainly used to resolve the relevant disease mechanisms and scientific and security issues of the treatment. OBJECTIVE: To summarize currently used experimental animal models of intervertebral disc degeneration study, and to dynamically observe and confirm the pathological process of disc degeneration based on disc imaging, morphology, biomechanics and biochemical changes. METHODS: Using “intervertebral disc degeneration, animal models, in vivo, in vitro” in English as the search words, Cochrane Library (No. 1 2009), Cochrane Library Database of Controlled Clinical Trials (No. 1 2009), MEDLINE from 1990 to March 2009, EMbase from 1990 to March 2009, Current Controlled Trials, and the National Research Register were retrieved. Literature was limited to English language. The disc imaging, morphology, biomechanical and biochemical composition and other indicators, as well as the pathological process of disc degeneration served as the evaluation indices. The articles related to the intervertebral disc cell culture models, the whole disc tissue culture model, mechanical model, injury model, biological model, genetically modified models, spontaneous models were included. The repetitive researches and those unrelated to animal models of intervertebral disc degeneration were excluded. RESULTS AND CONCLUSION: The establishment of a reliable animal model can provide favorable conditions for studying the pathogenesis of intervertebral disc degeneration, at the same time, provides a good experimental vehicle for various researches about the repair treatment of intervertebral disc degeneration. Animal models of intervertebral disc degeneration can be divided into two categories: in vitro models and in vivo models of disc degeneration and repair. The former can be assigned into disc cell culture models and whole disc tissue culture model; the latter is assigned into mechanical models, injury models, biological models, genetically modified models, spontaneous models and so on. The above models are commonly used in the study of the occurring mechanism of disc degeneration, as well as the feasibility and effectiveness of a variety of treatments. However, there is still no generally accepted animal models as an ideal disc degeneration model, various types of models reported have their own advantages and disadvantages. Related Articles | Metrics

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Theoretic study and preparing characteristics of animal pulmonary hypertension models Li Bo, He Wei 2010, 14 (11):  2039-2042.  doi: 10.3969/j.issn.1673-8225.2010.11.035 Abstract ( 366 )   PDF (455KB) ( 727 )   Save BACKGROUND: The establishment of animal pulmonary hypertension models will promote the improvement of clinical diagnosis and treatment on pulmonary hypertension. OBJECTIVE: To analyze and summarize the establishment of animal pulmonary hypertension models. METHODS: The databases of PubMed and CNKI were retrieved with key words of “pulmonary hypertension and animal model”. Studies concerning the following aspects were included: using surgery method to prepare animal pulmonary hypertension models; using drug injection method to prepare animal pulmonary hypertension models; animal severe pulmonary hypertension model; pathophysiological comparison between animal models and pulmonary hypertension patients; and drug intervention on animal pulmonary models.  RESULTS AND CONCLUSION: The methods for animal pulmonary hypertension models construction including: Surgical shunt, monoerotatine injection, chronically hypoxic and so on, in addition, the animal several severe pulmonary hypertension models which appeared at distal pulmonary artery neointimal lesion formation was reported. Through continuous improvement, they have provided insights into pathological cellular and molecular signaling pathways. However, how closely any of the models simulate to the cellular and molecular pathobiology of human pulmonary hypertension was poorly understood. Related Articles | Metrics

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Role of placenta-derived growth factor in vascular regeneration of ischemic cerebrovascular diseases Wu Lei, Luo Yong 2010, 14 (11):  2043-2047.  doi: 10.3969/j.issn.1673-8225.2010.11.036 Abstract ( 382 )   PDF (414KB) ( 449 )   Save BACKGROUND: The effect of placenta-derived growth factor (PLGF) on promoting vascular regeneration remains uncertain. In studies of models with PLGF deletion, PLGF has been considered to play critical role in inducing angiogenesis. OBJECTIVE: To summarize the research advances in effect of PLGF and its receptor in vascular regeneration and ischemic cerebral disease. METHODS: A computer-based online search of Pubmed and CNKI was performed for relevant articles published between 1990 and 2009 with key words of “PLGF” in Chinese and “PLGF, angiogensis” Articles related with biological characteristics of PLGF and its roles in vascular regeneration and articles related with angiogenesis and PLGF expression after cerebral ischemia were selected. Repetitive articles were excluded. RESULTS AND CONCLUSION: The role of PLGF and its receptor in vascular regeneration, mechanism of PLGF and its receptor to promote angiogenesis, vascular endothelial growth factor A, and the expression of PLGF and its receptor in brain after cerebral ischemia were introduced. PLGF can promote pathological angiogenesis, arteriogenesis, branch formation and hemopoietic progenitor cell mobilization. PLGF and its receptor play an important role in the angiogenesis of ischemic cerebral disease and display a good clinical prospect in the treatment of ischemic cerebral disease. However, the specific role and mechanism require further study. Related Articles | Metrics

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Morphological changes in cervical vertebral body patients with osteoporosis and cervical spondylosis: Case-control study Song Hai-feng, Tian Rong, Xia Gang, Zhang Xue-li, Xu Tian-tong, Shen Qing-feng 2010, 14 (11):  2048-2051.  doi: 10.3969/j.issn.1673-8225.2010.11.037 Abstract ( 340 )   PDF (307KB) ( 470 )   Save BACKGROUND: Some scholars have found that cervical vertebral body bone trabecula was reduced, became thin, even perforated in old patients with osteoporosis. Whether this change will induce cervical vertebral body deformation, and what relationship to the onset of cervical syndrome. OBJECTIVE: To study the relation of cervical spondylotic myelopathy and osteoporosis by measuring and comparing. METHODS: A total of 40 subjects with normal lumbar vertebra density and without cervical spondylosis were enrolled as control group, averagely 32.5 years. A total of 30 patients with cervical spondylosis served as cervical spondylosis group, averagely 43.6 years. Totally 46 patients with cervical spondylosis and osteoporosis served as combined with osteoporosis group, averagely 58.6 years. 116 subjects underwent radiograph. Height and sagittal diameter of the vertebral body, ratio of height to sagittal diameter of the vertebral body, and ratio of sagittal diameter of cervical canal/ vertebra body were measured. RESULTS AND CONCLUSION: Compared with the control group, vertebral height was decreased, and sagittal diameter became longer (P < 0.05), and the ratio of sagittal diameter of cervical canal/ vertebra body became smaller (P < 0.05) in the combined with osteoporosis group. Vertebral body deformation was characterized by decreased vertebral height and prolonged sagittal diameter became flat. Results suggested that osteoporosis induced cervical vertebral deformation, correlation between osteoporosis and cervical spondylosis, which may be a factor for cervical spondylosis development Related Articles | Metrics

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Normal meniscus and discoid meniscus injuries: Classification and tissue repair under an arthroscopy Huang Yuan-xia, Duan Yong-zhuang, Xu Hai-bin 2010, 14 (11):  2052-2055.  doi: 10.3969/j.issn.1673-8225.2010.11.038 Abstract ( 360 )   PDF (365KB) ( 723 )   Save BACKGROUND: Meniscus plasty and prosthesis under an arthroscopy are widely used in treating discoid meniscus or normal meniscus injury. The normal shapes of meniscus are maintained by excising medial partial tears and suturing the lateral partial tears. OBJECTIVE: Injury classification and special features of discoid meniscus and normal meniscus were observed, and corresponding treating methods were used, to discuss the therapeutic effect of repair.  METHODS: Totally 260 cases of meniscus injury in knee joint were selected from the First Affiliated Hospital of Xinxiang Medical University from September 2003 to June 2008. They were divided into normal meniscus injury group (n=215), and discoid lateral meniscus injury (n=45). People with complete clinical data before operation and following data, confirmed by arthroscopy were selected. Meniscus injury in 260 cases was examined by arthroscopy, and plasty, or partial excision or incomplete resection or full resection was performed according to the injury types and surgical circumstance. RESULTS AND CONCLUSION: All subjects were followed for half a year to 2 years. The knee function was evaluated by the Tenger criterion. Four tear types of Discoid meniscus were as follows: 73.3% horizontal, 15.5% complex, 4.4% radial, and 2.2% longitudinal. Five tear types of normal meniscus were as follows: 47.9% longitudinal, 28.8% radial, 11.6% horizontal, 6.5% transverse and 5.1% complex. In 215 cases of normal meniscus, there were 165 knees with excellent function, 35 with good, 8 with fair and 7 with poor. The excellent and good rate was 93.0%. In 45 cases of discoid meniscus, there were 25 knees with excellent function, 16 with good, 3 with fair and 1 with poor. The excellent and good rate was 91.1%. Discoid meniscus is different from normal meniscus in anatomical feature and histological structure, therefore, the arthroscopic surgery for discoid meniscus is also different. The most function of meniscus should be remained and the degeneration of knee joint should be delayed. Meniscus plasty should be performed in all discoid meniscuses except the patients with severe tear of meniscus. Related Articles | Metrics

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Investigation of pulse pressure and related factors of 1 989 healthy adults in Sichuan province Xu Dong-jiang, Si Qin, Zu Shu-yu, Xu Tao, Han Shao-mei, Xu Cheng-li, Zhu Guang-jin 2010, 14 (11):  2056-2059.  doi: 10.3969/j.issn.1673-8225.2010.11.039 Abstract ( 310 )   PDF (361KB) ( 529 )   Save BACKGROUND: High pulse pressure is the signal of arteriosclerosis or cardiovascular diseases for both hypertension and normal blood pressure population. OBJECTIVE: The pulse pressure and related factors of healthy adults in Sichuan province were analyzed to explore the method for preventing excessive pulse pressure and improving the quality of life. METHODS: Hierarchical, cluster sampling were performed in 1 989 healthy adults. The physical examination, blood glucose, renal function and blood fat were detected, and the relation of pulse pressure and its related factors were analyzed. RESULTS AND CONCLUSION: Pulse pressure increased following the age, especially showed a linear increasing in those who more than 50-year-old. The increased pulse pressure leaded to larger fasting plasma glucose, uric acid, blood urea nitrogen, total cholesterol, triacylglycerol, systolic pressure and diastolic pressure, in addition, the age, body mass index, waist-to-hip ratio, triacylglycerol and pulse pressure showed a linear relationship. But physical exercise was negative correlated to pulse pressure. With pulse pressure increasing, the renal function and regulating capacity of blood glucose was decreased. The results suggested that mid-aged population should take reasonable diet and more physical exercise to prevent central obesity and excessive pulse pressure, reduce the risk of renal dysfunction and the incidence of cardiovascular disease. Related Articles | Metrics

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Morphological characteristics of Chinese excellent men cyclists in 1-km time trials Zhou Qiang 2010, 14 (11):  2060-2063.  doi: 10.3969/j.issn.1673-8225.2010.11.040 Abstract ( 289 )   PDF (315KB) ( 335 )   Save BACKGROUND: Previous studies with regard to male track short-distance cyclists are concentrated on the descriptive and comparison studies of morphological characteristics of athletes. Up to now, few studies report the relationships between morphological characteristics and sports performance in men 1-km cyclists. OBJECTIVE: To explore an effective morphological index for evaluating physical stamina level of men cyclists in 1-km time trials, and to establish morphological models and evaluation standards for men cyclists in 1-km time trials in China. METHODS: Totally 17 men who had prepared for the 1-km cycling time trials of the fifth cycling city sport meeting in Taiyuan were examined. With the methods of step by step regression, gray correlation, measurement and evaluation theory, the morphological characteristics of these excellent cyclists were explored, and utilized in establishing morphological models, single-grading table and the ranking evaluation criteria. RESULTS AND CONCLUSION: The typical indexes of excellent 1-km men cyclists were body height and fat-free body weight. The morphological characteristics of these excellent cyclists were as follow: tall, long leg and Achilles tendon; high foot arch; heavy body weight and fat-free body weight; as well as thick thigh. The present study established the evaluation model of the morphological characteristics and evaluation standards for 1-km men cyclists.  Related Articles | Metrics

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Effect of basic fibroblast growth factor on the gene expression of decorin in periodontal ligament cells Cheng Yuan, Zeng Zhao-fang, Zhu Jun 2010, 14 (11):  2068-2071.  doi: 10.3969/j.issn.1673-8225.2010.11.042 Abstract ( 266 )   PDF (277KB) ( 492 )   Save OBJECTIVE: Studies have confirmed that basic fibroblast growth factor stimulating periodontal ligament cells can promote the proliferation of human periodontal ligament cells, so as to facilitate the reconstruction of lost periodontal tissues. By use of different concentrations of basic fibroblast growth factor, the in vitro cultured normal human periodontal ligament cells were stimulated to observe the decorin gene expression in the periodontal ligament cells. METHODS: The periodontal ligament cells were isolated and cultured using trypsin digestion method. The third generation of cells at logarithmic growth phase were preserved in DMEM containing 10% DMSO and 20% FBS frozen cryopreservation fluid, then moved into liquid nitrogen preservation at the second day. Immunohistochemistry was used to identify anti-vimentin and cytokeratin staining. The sixth generation of human periodontal ligament cells were divided into experimental group and control group. The experimental group was cultured in DMEM culture medium containing basic fibroblast growth factor at the concentration of 0.1, 1, 10 μg/L under standard conditions for 24 hours; control group was cultured with DMEM culture medium under standard conditions for 24 hours. The intracellular decorin gene expression was determined using RT-PCR. RESULTS: By microscopic observation, the cells in the control group had no significant changes, and cells proliferated in the experimental group, before stimulation the cells at the bottom of the bottle became denser than those in sparse regions. Adding basic fibroblast growth factor into periodontal ligament cells, could significantly decrease the mRNA expression of decorin, along with the increase of basic fibroblast growth factor concentration, the inhibition effect gradually weakened and became the strongest in the 0.1 μg/L but the weakest at 10 μg/L. CONCLUSION: The basic fibroblast growth factor stimulation can promote the proliferation of periodontal ligament cells and inhibit the synthesis of decorin in a dose-dependent manner, 0.1 μg/L induces the strongest inhibitory effect. Related Articles | Metrics

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Effects of tumor necrosis factor-alpha on maturity of mouse embryonic fibroblasts Yang Wen-jiu, Zou Yun-wen, Wang Zhi-jie 2010, 14 (11):  2072-2075.  doi: 10.3969/j.issn.1673-8225.2010.11.043 Abstract ( 352 )   PDF (382KB) ( 491 )   Save BACKGROUND: In recent years, studies have shown the effects of tumor necrosis factor-α (TNF-α) on fibroblasts at different organizations in a tissue-specific and dose-dependent manner. OBJECTIVE: To investigate the biologic effects of TNF-α and its signal transduction pathway-specific kinase inhibitors on mouse embryonic fibroblasts (NIH3T3). METHODS: NIH3T3 cells were cultured in vitro, and then the cells were assigned into 3 groups. Cells in the control group were cultured in DMEM high-glucose medium with 2% serum; those in the TNF-α group were cultured in 100 μg/L TNF-α medium; those in the TNF-α+Anti-TNFRSF1B group were firstly cultured in medium with 50 μg/L Anti-TNFRSF1B for 1 hour, and then placed in the medium with 100 μg/L TNF-α. RT-PCR and Western blot methods were used to evaluate mRNA and protein expressions of type Ⅰ collagen and matrix metalloproteinase 3 (MMP3) in each group. RESULTS AND CONCLUSION: In this experiment, NIH3T3 cells cultured with a certain concentration of TNF-α, the specificity kinase signal of transduction pathway presented with phosphorylation or protein activation, and the signal pathway was activated, which promoted MMP3 activation, and significantly reduced the expression of type Ⅰ collagen. The effect of TNF-α was certainly inhibited, but not completely eliminated after adding its signal transduction pathway inhibitor Anti-TNFRSF1B. This further proves the role of TNF-α on NIH3T3 activation. Related Articles | Metrics

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Cell activity of human de-epidermized dermis and its characteristics of tissue structure Lu Hong-guang, Dong Dan, Ma Yue-hong, Guo Zhe, Cui Shao-shan, Wang Ya-kun, Chen Hong-duo 2010, 14 (11):  2076-2080.  doi: 10.3969/j.issn.1673-8225.2010.11.044 Abstract ( 347 )   PDF (346KB) ( 589 )   Save BACKGROUND: Study confirmed that the de-epidermized dermis (DED) can be used as dermal substitute and may form epidermal structure after incubating keratinocytes. However, the cell biological activity, tissue structure characteristics and the basement membrane component analysis of dermal substitute have been reported less. OBJECTIVE: To investigate the cell activity and the tissue structure characteristics of DED. METHODS: Skin flap was treated with 56 ℃ phosphate buffered solution to remove the epidermis, and the dermal cell components were deleted by freezing and thawing with liquid nitrogen to obtain DED. The DED cell activity was detected with tissue culture method, hematoxylin nuclear staining was used to determine the DED cell nuclei, and vimentin immunohistochemistry was applied for fibroblast determinations. The basement membrane and its components were detected using Periodic Acid-Schiff staining and collagen type IV immunohistochemistry. Van Gieson stain, Weigart stain and those double staining were respectively used to determine DED collagen fibers and elastic fibers. The DED ultrastructure was observed under transmission and scanning electron microscope. RESULTS AND CONCLUSION: Using tissue culture method, the cultured DED did not exhibit cell growth at 2 weeks. Hematoxylin-eosin staining showed no nuclear in DED, vimentin immunohistochemistry showed no vimentin expressed in DED. Van Gieson staining showed DED collagen fibers were stained as rose red, Weigert staining showed DED elastic fibers were stained as purplish black, double staining further demonstrated uniform arrangement of collagen fibers and elastic fibers. DED surface and the remaining appendages were strongly positive for Periodic Acid-Schiff staining, and type IV collagen expression was significant. Transmission and scanning electron microscope results showed that, the DED elastic fibers and collagen overlap arranged with pore intervals, they intercrossed into a network. There is no living cell component in DED, dermal matrix surface and appending organ luminal wall still retain glycogen, type IV collagen and other basement membrane components, dermal matrix is rich in collagen and elastic fibers, it is a three-dimensional collagen matrix similar to in vivo dermis. Related Articles | Metrics

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Gene expression profile in osteoblastic differentiation of bone marrow stromal cells stimulated by simvastatin Gene chip analysis Meng Ya-qiang, Zhang Liu, Tian Fa-ming, Han Da-cheng, Zheng Jie, Cai Jun 2010, 14 (11):  2081-2085.  doi: 10.3969/j.issn.1673-8225.2010.11.045 Abstract ( 336 )   PDF (334KB) ( 462 )   Save BACKGROUND: Simvastatin enhanced the expression of bone morphogenetic protein-2 (BMP-2), which plays an anabolic role in bone metabolism and osteoblastic lineage differentiation. However, little is known about the molecular mechanism of simvastatin on regulation of bone marrow stromal cells differentiation.  OBJECTIVE: To investigated the effect of simvastatin on osteoblastic differentiation of bone marrow stromal cells based on genetics level. METHODS: Bone marrow stromal cells derived from femur and tibia were cultured in different mediums with simvastatin or vehicle for 7 days. Following extraction and purification, mRNA was reverse-transcripted into cDNA. Fluorescence labeling was employed and the samples were then hybridized with oligonucleotide chip to screen the different genes, which were utilized to analyze osteogenesis-related factors. Alkaline phosphatase and Von Kossa staining were performed at days 14 and 21, respectively. RESULTS AND CONCLUSIONS: At day 14, alkaline phosphatase-positive cells were more in the experimental group than control group. Von Kossa staining demonstrated that simvastatin could promote BMSCs osteoblastic differentiation and mineralization. Comparative analysis showed that 103 genes out of 22 575 rat genes had differential expression (≥ 2 fold or ≤ 0.5 fold), and some genes were related to cell proliferation and ostoeblastic differentiation, including C/EBPδ, Cited, Ascl2, Ptpn16, Wisp2, Tieg, etc. Simvastatin could induce osteoblastic differentiation of bone marrow stromal cells, involving in many osteogenetic-related genes. Related Articles | Metrics

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Expression and role of matrix metalloproteinases and tissue inhibitor of metalloproteinases in a rat model of traumatic deep vein thrombosis Zhang Yu-bing, Li Wen, Yao Li-qing, Zhao Xue-ling, Wang Bing, Li Hong-kun, Ning Ya, Song En 2010, 14 (11):  2086-2090.  doi: 10.3969/j.issn.1673-8225.2010.11.046 Abstract ( 273 )   PDF (314KB) ( 467 )   Save BACKGROUND: The molecular mechanism of traumatic deep vein thrombosis is complex. Numerous studies focus on clinical observation and epidemiology, but its molecular mechanism has not been a new breakthrough. OBJECTIVE: By use of gene array technology, this study was aimed to study the expression changes of matrix metalloproteinases in rat models of traumatic deep vein thrombosis, and to explore the roles of matrix metalloproteinases in traumatic deep venous thrombosis. METHODS: A total of 150 SD rats, SPF grade, of 8-12 weeks old, body weight of 250-300 g, were divided at random into normal control group (n=10) and model group (n=140). Rat traumatic deep venous thrombosis models were set up by clamping the femoral vein and fixing the bilateral hind limbs, and the fixation of hip spica with plaster bandage was conducted in each group. Then rats were divided into 7 subgroups: post-traumatic 0.5 hours, post-traumatic 2.5 hours (initial period of thrombosis), post-traumatic 25 hours (thrombogenesis at thrombotic crest-time), post-traumatic 25 hours (non-thrombogenesis at the thrombotic crest-time), post-traumatic 72 hours (thrombus resolution), post-traumatic 72 hours (thrombus insolution) and post-traumatic 168 hours (nonthrombosis). At the corresponding phasess, the femoral vein tissues were incised, and total RNA of femoral vein was extracted using Trizol one-step method. Applying Genechip Rat Genome 430 2. 0 genechips, the gene expressions in femoral vein were detected in different groups. The rate of traumatic deep venous thrombogenesis and non-thrombogenesis, the rate of thrombi solution and insolution were observed; the expressions of matrix metalloproteinases and tissue inhibitor of metalloproteinases at different time phases was detected by gene array data analysis. RESULTS AND CONCLUSION: Three model rats died and the remaining 147 rats were involved in the final analysis. At the post-traumatic 25 hours, the rate of thrombogenesis was 50.5% and nonthrombogenesis was 49.5%. To the post-traumatic 168 hours, the rate of thrombus solution was 56.7% and thrombus insolution was 43.3%. Both matrix metalloproteinases and tissue inhibitor of metalloproteinases exhibited differential expressions in the course of traumatic deep venous thrombosis. Under the thrombus insolution state, matrix metalloproteinases continued to show a high expression, tissue inhibitor of metalloproteinase expression was down-regulated in the thrombus formation, was significantly inhibited in the thrombus insolution process. In the process of traumatic deep vein thrombosis and insolution, matrix metalloproteinase was closely related to traumatic deep vein thrombosis, the matrix metalloproteinase/tissue inhibitor of metalloproteinases are likely to affect the biological state of thrombosis. Related Articles | Metrics