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11 March 2012, Volume 16 Issue 11 Previous Issue    Next Issue

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Effects of gene therapy on the expression of transforming growth factor beta 1 during mandible distraction osteogenesis    He Xiao-chuan, Li Shao-lan, Hu Chun-bing, Liu Zhen, Gao Zhi-dan, Yin Kang, Wu Guo-ping, Guo Li 2012, 16 (11):  1901-1905.  doi: 10.3969/j.issn.1673-8225.2012.11.001 Abstract ( 282 )   PDF (438KB) ( 335 )   Save BACKGROUND: Although recent studies have shown that the local gene therapy can promote the formation of new bone in distraction gap, the effect of gene therapy on the expression of local growth factors is unclear. OBJECTIVE: To investigate the effect of electroporation-mediated gene therapy on the expression of transforming growth factor-β1 (TGF-β1) during mandibular distraction osteogenesis in a rabbit model. METHODS: Mandibular distraction of 0.8 mm/d was performed in New-Zealand rabbits at 3 days after bilateral mandibular osteotomy and lasted for 7 days. After the completion of distraction, the rabbits were randomly divided into five groups. 2 μg      (0.1 μg/μL) recombinant plasmid pIRES-hVEGF165-hBMP2, pIRES-hBMP2, pIRES-hVEGF165, pIRES and normal saline was injected into the distraction area respectively. After injection, every group employed electroporation. RESULTS AND CONCLUSION: Immunohistochemical staining showed that TGF-β1 was mainly expressed in the cytoplasm. Seven days after transfection, the staining of TGF-β1 was positive on osteocytes of bone edge, osteocytes of woven bone callus and osteoblasts on the surface of bone callus. At 14 days, the expression of TGF-β1 was found on osteocytes of newborn woven bone callus, osteoblasts on the surface of bone callus, stromal cells in granulation tissue, mononuclear giant cells and multinucleated giant cells. At 28 days, the positive expression of TGF-β1 was decreased obviously. Compared with the injection of pIRES and normal saline, the expression of TGF-β1 was significantly higher after the injection of recombinant plasmid pIRES-hVEGF165-hBMP2, pIRES-hBMP2 and pIRES-hVEGF165 (P < 0.05 or P < 0.01). It suggests that gene therapy can promote TGF-β1 expression effectively and promote the formation of new bone and cell matrix in distraction gap. Related Articles | Metrics

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Effect of adiponectin on bone regeneration following rapid mandibular distraction osteogenesis in rabbits  Shui Xue-ping1, Jiang Xiao-wen2, Ye Bin1, Hu Jing1 2012, 16 (11):  1906-1909.  doi: 10.3969/j.issn.1673-8225.2012.11.002 Abstract ( 249 )   PDF (609KB) ( 404 )   Save BACKGROUND: Adiponectin participates in bone metabolism and angiogenesis, which is beneficial for bone growing. However, the effect of adiponectin on bone regeneration following distraction osteogenesis is not clear. OBJECTIVE: To investigate the effects of intermittent local administration of adiponectin on bone regeneration following establishment of rapid mandibular distraction models in rabbits. METHODS: Sixteen New Zealand white rabbits were randomly divided into experimental group and control group. Rapid unilateral mandibular distraction osteogenesis models were established with 2 mm/d distraction rate. At the 1, 3 and 5 days of the distraction, 200 μL of phosphate buffer solution or 200 μL of phosphate buffer solution containing 2 μg recombinant human adiponectin was intermittently injected into the distraction gap of the control and experimental groups, respectively. RESULTS AND CONCLUSION: Bone regeneration was found in the distracted callus from both groups. Histological observation and micro-CT examinations confirmed that the new bone formation and mineralization of the distraction callus in experimental group was significantly greater than that of the control group. Intermittent administration of adiponectin can markedly promote the bone regeneration in rabbit rapid mandibular osteodistraction. Related Articles | Metrics

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Effects of recombinant human bone morphogenetic protein combined with basic fibroblast growth factor on the repair of articular cartilage defects Zhu Guo-hua1, Cai Jian-ping1, Guo Cui-ling1, Liao Jia-xin1, Liu Yong1, Luo Hong-tao1, Xu Guo-hua2, Hu Hong-tao2 2012, 16 (11):  1910-1914.  doi: 10.3969/j.issn.1673-8225.2012.11.003 Abstract ( 324 )   PDF (459KB) ( 390 )   Save BACKGROUND: The synergy of various cell growth factors attracts more and more attention in the course of cartilage metabolism. However, there are few reports of repairing cartilage defects with combined cell growth factors, and the effect remains unknown at present. OBJECTIVE: To study the repairing effect of recombinant human bone morphogenetic protein (rhBMP) combined with basic fibroblast growth factor (bFGF) on articular cartilage defects. METHODS: After the model of articular cartilage defects was made, 24 Japan big-eared white rabbits were randomly divided into four groups for intervention: rhBMP combined with bFGF (group A), single rhBMP (group B), single bFGF (group C), the fourth group was without injection and just filled with gelatin sponge (group D). RESULTS AND CONCLUSION: In general observation, articular cartilage defects were basically repaired but slightly uneven in group A, and there were no defects entirely repaired in groups B and C. In group D, articular cartilage defects were not repaired. The number of cartilage cells in group A was more than that in groups B, C and D (P < 0.05). The positive number of type Ⅱ collagens by immunohistochemical staining in group A was higher than that in the other groups. It indicated that rhBMP combined with bFGF could accelerate the formation of new articular cartilage, and had better effect than rhBMP or bFGF alone.  Related Articles | Metrics

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Dexamethasone effects on the proliferation of nucleus pulposus cells and the expression of type Ⅱ collagen-2 in rabbits Zhou Xiao-rui, Liu Shi-qing, He Bin, Zhang Nu 2012, 16 (11):  1915-1918.  doi: 10.3969/j.issn.1673-8225.2012.11.004 Abstract ( 250 )   PDF (368KB) ( 296 )   Save BACKGROUND: Dexamethasone is a common drug for cell proliferation. However, it remains unclear whether dexamethasone can promote the proliferation of nucleus pulposus cells. OBJECTIVE: To investigate the effects of dexamethasone on the proliferation of nucleus pulposus cells in rabbit intervertebral disk and the expression of type Ⅱ collagen secreted by nucleus pulposus cells. METHODS: The rabbit intervertebral disk was extracted in sterile processing. After conventional separation and digestion, nucleus pulposus cells were cultured and passaged for 7 days, and then dexamethasone with the concentration of 1, 10, 100,    1 000, 10 000 nmol/L was added. After that, the nucleus pulposus cells were cultured for 1-5 days, and the absorbance values at 490 nm of the cells were detected by 3-(4, 5)-dimethylthiahiazo (-z-y1)-3, 5-di-phenytetrazoliumromide (MTT) assay every day and the growth curve was drawn. In addition, 100 nmol/L dexamethasone was selected for culture of nucleus pulposus cells, and the nucleus pulposus cells cultured with no dexamethasone was as controls. RESULTS AND CONCLUSION: MTT assay results showed that dexamethasone had the proliferative effects on the cultured nucleus pulposus cells. The optimal action time was 48 hours and the optimal action concentration was 100 nmol/L. Reverse transcription-PCR results showed that dexamethasone could up-regulate the mRNA expression of type Ⅱ collagen which was significantly higher than that in the control group (P < 0.05). These suggest that dexamethasone can promote the proliferation of nucleus pulposus cells of rabbit intervertebral disk and up-regulate the expression of type Ⅱ collagen in rabbit nucleus pulposus cells. Related Articles | Metrics

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Effects of basic fibroblast growth factor on adenosine triphosphatase activity and antioxidation of alcoholism rat models    Huang Jun-jie, Wang Cai-bing, He Xian-jiao, Huang Li-juan, Huang Yan-feng, Liang Zuo-ren, Yang Xiu-ying, Li Yun, Zhao Shan-min 2012, 16 (11):  1919-1922.  doi: 10.3969/j.issn.1673-8225.2012.11.005 Abstract ( 294 )   PDF (339KB) ( 282 )   Save BACKGROUND: Basic fibroblast growth factor (bFGF) has multiple biological activities, and has positive effect on the recovery of tissue trauma. But the effect of bFGF against alcoholism has not been reported. OBJECTIVE: To study the effect of bFGF on the activities of adenosine triphosphatase (ATPase) and superoxide dismutase (SOD) and malondialdehyde (MDA) in cerebral cortex and liver tissue of alcoholism rat model. METHODS: The alcoholism rat models were established by perfusing stomach with alcohol. Thirty Wistar rats were randomly divided into three groups. Normal saline group and bFGF treatment group were injected with normal saline and bFGF, respectively, at 60 days after modeling; Alcoholism model group was treated with no intervention. Another 10 rats without alcohol served as control group. RESULTS AND CONCLUSION: The activities of ATPase and SOD in the cerebral cortex and liver tissue of alcoholism model group were significantly decreased than those in the control group, but the content of MDA was significantly higher than that in the control group (P < 0.05). After bFGF intervention, the activities of ATPase and SOD in the cerebral cortex and liver tissue were significantly increased compared with the normal saline group and alcoholism model group, but the content of MDA was significantly decreased (P < 0.05-0.01). bFGF can increase the activities of ATPase and the antioxidative ability of brain tissue in alcoholism model rats. The bFGF can protect the brain and liver of rats against alcoholism.   Related Articles | Metrics

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Effects of Wenyang Huoxue recipe on transforming growth factor beta/Smad signal transduction pathway in model rats of renal interstitial fibrosis  Hu Shao-jin1, Li Jun2 2012, 16 (11):  1923-1926.  doi: 10.3969/j.issn.1673-8225.2012.11.006 Abstract ( 270 )   PDF (456KB) ( 368 )   Save BACKGROUND: Transforming growth factor β/Smad signal transduction pathway plays an important role in the occurrence and development of renal fibrosis. OBJECTIVE: To investigate the effects of Wenyang Huoxue recipe on transforming growth factor β/Smad signal transduction pathway in renal interstitial fibrosis rats. METHODS: A total of 28 male SD rats of clean grade were randomly assigned into sham operation group and model group. Left urethral ligation was performed in rats to construct unilateral ureteral obstruction model. The left ureter of sham operation rats was isolated without ligation after opening the abdominal cavity; then the abdominal cavity was closed. The rats were administrated with Wenyang Huoxue recipe by gavage once. Rats in both groups were administered by gavage for 14 days. RESULTS AND CONCLUSION: The expression of transforming growth factor β1 and Smad3 in the Wenyang Huoxue group was significantly lower than that in the model group (P < 0.01). The expression of Smad7 in the Wenyang Huoxue group was significantly larger than that in the model group (P < 0.01) and similar to that in the fosinopril group. It is conjectured that Wenyang Huoxue recipe may inhibit the expression of transforming growth factor β1 and Smad3, increase Smad7 expression, thereby affecting the transforming growth factor β/Smad signaling pathway, and thus play a role in the prevention and cure of renal interstitial fibrosis. Related Articles | Metrics

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Construction and expression of human vascular endothelial growth factor 165 gene lentiviral expression vector Yi Chao, Wang Xi-yan, Li Hai-jun, Wang Hai 2012, 16 (11):  1927-1932.  doi: 10.3969/j.issn.1673-8225.2012.11.007 Abstract ( 264 )   PDF (479KB) ( 389 )   Save BACKGROUND: Studies have confirmed that vascular endothelial growth factors (VEGF) play an important role in the course of normal liver regeneration, while, there are very few reports on the same role in the cirrhotic liver nationally and internationally. OBJECTIVE: To construct the lentivirus vector containing human VEGF165 gene, and to examine the expression of VEGF165 in normal rat liver cells after transfecting BLR 3A rat liver cells, in order to lay a foundation for the study about the effects of VEGF165 on the regeneration power of remnant liver tissue after hepatectomy in the rats with cirrhosis. METHODS: The VEGF165 gene was cloned to a lentiviral expression vector by recombinant DNA technology. The positive clones were screened, and lentiviral packaged systems (ViraPowerTM Packaging Mix) were co-transfected to package virus in 293T cells by lipofection with target gene plasmid. Real-time PCR technique was used o test the titer of pLenti6/V5-D-TOPO-VEGF165. The BLR 3A rat liver cells were transfected by pLenti6/V5-D-TOPO-VEGF165 and the expression of VEGF165 mRNA and protein was detected by reverse transcriptase-PCR and Western blot technique. RESULTS AND CONCLUSION: The lentivirus vector containing VEGF165 was successfully constructed. Virus titer could reach as high as 1.18×107 VP/mL. The transfer efficiency of green fluorescent protein was more than 80% in the BLR 3A rat liver cells after transfection with pLenti6/V5-D-TOPO-VEGF165 for 72 hours. Reverse transcriptase-PCR and Western blot results showed that the VEGF165 gene expression of the transfected group was positive. Lentivirus vector pLenti6/V5-D-TOPO-VEGF165 could transfect BLR 3A rat liver cells effectively, and VEGF165 gene was expressed after transfection.   Related Articles | Metrics

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Interaction of cardiac transcription factor Nkx2.5 and GATA4 Ding Jian-dong1, Fang Xiang1, Li Kai-ru2, Yao Yu-yu1, Tao Shao-yu1, Wang Dian1, Ma Gen-shan1 2012, 16 (11):  1933-1936.  doi: 10.3969/j.issn.1673-8225.2012.11.008 Abstract ( 326 )   PDF (295KB) ( 439 )   Save BACKGROUND: Heart development results from the interaction among various factors. Research on the interaction of Nkx2.5 and GATA4 may help us to further understand the process of heart development. OBJECTIVE: To research the interaction of cardiac transcription factor Nkx2.5 and GATA4 in vitro. METHODS: On the basis of the construction of plasmids of pCMV-Myc-GATA4, pCMV-HA-Nkx2.5 and pTal-luc-BNP, plasmids were transfected into COS7 cells. The interaction of Nkx2.5 and GATA4 was identified by co-immunoprecipitation and luciferase analysis. The possible synergistic activation of Nkx2.5 and GATA4 on BNP promoter sequence was analyzed by luciferase.  RESULTS AND CONCLUSION: Co-immunoprecipitation experiments showed that Nkx2.5 and GATA4 could be precipitated by each other, demonstrating that Nkx2.5 and GATA4 could directly interact with each other in COS7 cells. The pTal-luc-BNP combined with Nkx2.5 and GATA4 were co-transfected into COS7 cells. Compared with single transfected with Nkx2.5 or GATA4 plasmid, co-transfected with Nkx2.5 and GATA4 displayed synergistic transcriptional activation of the BNP promoter. Nkx2.5 and GATA4 play important roles separately in the process of heart development, and the interactions of Nkx2.5 and GATA4 are also vital in normal heart development. Related Articles | Metrics

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Effect of joint synovial fluid on biodynamics and histology of tendon-bone healing after reconstruction of anterior cruciate ligament in rabbits Dong Xiang-hui, Ling Ming, Feng Wei-lou, Tian Xin 2012, 16 (11):  1937-1940.  doi: 10.3969/j.issn.1673-8225.2012.11.009 Abstract ( 297 )   PDF (332KB) ( 527 )   Save BACKGROUND: At present, effect of joint synovial fluid on the strength of graft and tendon-bone healing after reconstruction of anterior cruciate ligament (ACL) is inconclusive. OBJECTIVE: To observe the effect of joint synovial fluid on biodynamics and histology of tendon-bone healing after reconstruction of ACL in the rabbits. METHODS: Semitendinosus tendons were taken from lower extremities of New Zealand white rabbits to reconstruct ACL simulation synovial fluid influence model with ipsilateral semitendinosus tendons, and contralateral semitendinosus tendons were used for implanting “U”-shaped on the femoral condyle to avoid the influence of the synovial fluid. Specimens were collected at 4 weeks after operation for biomechanical measurement and histological observation. RESULTS AND CONCLUSION: Biomechanical measurement at 4 weeks after reconstruction showed that the average load of “U” shaped implanted tendon breakage was significantly higher than that of the ACL reconstructed tendon (P < 0.01). Histological observation showed that necrotic tendon tissues in bone tunnel were replaced by new bone tissues and fibrous tissues. Sharpey’s fibers and fibrocartilages were appeared on the tendon-bone interface. The number of bone cells on the tendon bone interface of the “U” shaped implanted tendon was significantly higher than that of ACL reconstructed tendon (P < 0.01), superior to intra-articular group. It results showed that biomechanics and tendon bone healing of the “U” shaped implanted tendon were both superior to the ACL reconstructed tendon. It is indicated that joint synovial fluid of ACL reconstruction has an adverse effect on ligament strength and tendon bone healing. Related Articles | Metrics

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Plantar pressure characteristics of migrant workers who perform physical labor   Zhang Qing-lai1, Tian Xin-ming2, Guo Qiang3 2012, 16 (11):  1941-1944.  doi: 10.3969/j.issn.1673-8225.2012.11.010 Abstract ( 362 )   PDF (247KB) ( 346 )   Save BACKGROUND: Researches on plantar pressure characteristics at home and abroad are focused on ordinary people and patients. The plantar pressure characteristics of migrant workers are rarely reported. OBJECTIVE: To test the plantar pressure of migrant workers performing frequent physical labor. METHODS: A total of 30 migrant workers performing frequent physical labor were assigned into migrant worker group. While another 30 university students were assigned into university student group. Dynamic plantar pressures of all the subjects during natural walking were tested by a plate plantar pressure test system (Footscan USB2, Belgium). RESULTS AND CONCLUSION: The peak pressure of the first toe, the second and third metatarsus in the right foot of migrant workers was significantly higher than that of the university students (P < 0.01). There was no significant difference in foot arch index between the two groups (P > 0.05).The angle of foot deviation and the occurrence of strephenopodia in migrant worker group were larger than that in the university student group during natural walking (P < 0.01). These results demonstrate that frequent physical labor has an obvious effect on plantar pressure distribution in migrant workers. The phenomena include plantar pressure moving forward, increased strephenopodia extent and magnified foot deviation angle during natural walking. Therefore the possibility of lower limb injury becomes more likely. Related Articles | Metrics

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Biomechanics of rectus femoris tendon versus anterior cruciate ligament Zhu Xing-fei1, Zhang Xin-chao1, Chen Bo2 2012, 16 (11):  1945-1947.  doi: 10.3969/j.issn.1673-8225.2012.11.011 Abstract ( 309 )   PDF (230KB) ( 423 )   Save BACKGROUND: Rectus femoris tendon is on important substitute of anterior cruciate ligament reconstruction. OBJECTIVE: To compare the biomechanics of rectus femoris tendon and anterior cruciate ligament. METHODS: The rectus femoris tendon and anterior cruciate ligament specimen were obtained within postmortem 9.0-10.0 hours by anatomy, and the specimens were wrapped by saline moistened paper towels immediately, then sealed and placed in a -20 ℃ refrigerator for storage. The specimens were removed from the refrigerator before experiment and each specimen was cut into two samples, 10 rectus femoris tendon specimens and 10 anterior cruciate ligament specimens in each group. RESULTS AND CONCLUSION: The unit moludus of rectus femoris tendon was 63% of anterior cruciate ligament, the unit maximum load of rectus femoris tendon was 63% of anterior cruciate ligament. Rectus femoris tendon was suitable for anterior cruciate ligament substitute. Related Articles | Metrics

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Protective effect of sodium ferulate preconditioning on anoxia-reoxygenation injury of myocardial cells in adult rats Zeng Liang, Liu Ji-chun 2012, 16 (11):  1948-1953.  doi: 10.3969/j.issn.1673-8225.2012.11.012 Abstract ( 303 )   PDF (585KB) ( 491 )   Save BACKGROUND: Sodium ferulate preconditioning can protect myocardial cells, which has been proved in the experiments of isolated rat heart and neonatal rat myocardial cells, but there are few experimental studies regarding adult rat myocardial cells. OBJECTIVE: To explore the protective effect of sodium ferulate preconditioning on myocardial cells following anoxia-reoxygenation (A/R) in adult rats as well as its mechanism underlying adenosine triphosphate-sensitive potassium (K+ATP) channel . METHODS: Rat heart was perfused with Langendorff system. The purified adult rat myocardial cells were isolated by collagenase digestion method and then treated with simulated A/R solution, sodium ferulate (SF) as well as glibenclamide (Glib), a K+ATP channel blocker. Then, the myocardial cells were randomly divided into six groups: control group, A/R group, anoxia preconditioning (APC)+A/R group, SF+A/R group, Glib+APC+A/R group and Glib+SF+A/R group. RESULTS AND CONCLUSION: ①Myocardial cell viability: compared with the control group, the cell viability in the A/R group was significantly decreased (P < 0.01). Compared with the A/R group, the cell viability was significantly increased in the APC+A/R group and SF+A/R group (P < 0.01). The cell viability in the Glib+APC+A/R and Glib+SF+A/R groups had no difference from that of the A/R group, but significantly higher than that of the APC+A/R group (P < 0.01). ②Lactate dehydrogenase activity: the comparison results of lactate dehydrogenase activity were consistent with those of myocardial cell viability among these groups. ③Ultrastructure observation under a transmission electron microscope: the myocardial cells subjected to A/R injury were characterized by mitochondrial swelling, disappearance or deformation of mitochondrial cristae, disruption of nuclear membrane, and nuclear condensation. While the ultrastructure of the myocardial cells in the APC+A/R group and SF+A/R group were changed slightly, exhibiting regular mitochondria with uniform size, intact mitochondrial cristae, intima and extima as well as complete nuclear membrane. The injury was obviously reduced in the APC+A/R group and SF+A/R group compared with the A/R group. The cellular structures in the Glib+APC+A/R group and Glib+SF+A/R group were similar to those in the A/R group. These findings showed that SF preconditioning is effective in protecting the myocardial cells of adult rats from A/R injury and the cardioprotective effect of SF preconditioning is related to the activation of intracellular K+ATP channel.   Related Articles | Metrics

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Construction of short hairpin RNA lentiviral vector targeting rabbit Annexin A1 gene and identification of RNA interference efficiency  Liang Bo-wei1, Pan Xin-yuan2, Zhao Jin-min1, Yin Guo-qian2, Hu Feng1 2012, 16 (11):  1954-1958.  doi: 10.3969/j.issn.1673-8225.2012.11.013 Abstract ( 392 )   PDF (532KB) ( 493 )   Save BACKGROUND: Annexin A1 (ANXA1) protein is involved in the regulation of cell apoptosis. OBJECTIVE: To construct a short hairpin RNA (shRNA) lentiviral vector targeting rabbit ANXA1 gene and to detect its efficiency of gene silence in osteoblasts. METHODS: Targeting rabbit ANXA1 gene sequences, a pair of complementary small hairpin RNA (shRNA) oligonucleotides were designed, synthesized, annealed and cloned into pGLV/H1/GFP plasmid digested by BamHI and EcoRI to construct a vector named pGLV-shANXA1. The recombinant plasmid was identified by PCR and DNA sequencing. 293FT cells were co-transfected with pGLV-shANXA1, pGLV/helper-1,pGLV/helper-2 and pGLV/helper-3 to package into lentivirus particles named LV-shANXA1.The titer of virus was tested according to the expression level of GFP. The lentivirus particles were then transmitted into rabbit osteoblasts and its infection efficiency was assessed by flow cytometry. RESULTS AND CONCLUSION: The PCR identification and DNA sequencing showed that the fragment and nucleotides were in accordance with the target sequence and the shRNA sequence was successfully inserted into the pGLV/H1/GFP vector. The titer of concentrated LV-shANXA1 was 3.8×108 TU/L. The infection efficiency of lentiviral particles was 80% at multiplicity of infection (MOI) of 50 and 95% at MOI of 100 in osteoblasts after puromycin selection. Real-time PCR and Western blot results showed that the expression of ANXA1 in LV-shANXA1-901 infected osteoblasts were lower than that in the blank control, negative control, and other LV-shANXA1 infected cells (P < 0.05), it could achieve 71.2% interference efficiency at MOI of 50. The results demonstrate a lentiviral shRNA expression vector targeting the ANXA1 gene is successfully constructed and it can knockdown the expression of ANXA1 in osteoblasts. Related Articles | Metrics

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Culture in vitro and identification of human cervical carcinoma cells Ma Xiu-ping, Cheng Jing-xin, Zhang Yu, Yuan Min 2012, 16 (11):  1959-1962.  doi: 10.3969/j.issn.1673-8225.2012.11.014 Abstract ( 351 )   PDF (368KB) ( 367 )   Save BACKGROUND: There are individual differences in the patients suffered tumor. Therefore the research on the cancer line can not show accurate differences between individuals, while primary cell culture can show more similar characteristics to in vivo state. OBJECTIVE: To establish a method in vitro for primary culture of cervical carcinoma cells. METHODS: Cervical cancer cells were obtained from 23 fresh cervical cancer tissues using the trypsin and collagenase type Ⅰdigestion, and their growth curve was detected. Cell surface markers of CK17and CD44 were identified with immunofluorescence staining. RESULTS AND CONCLUSION: Cervical cancer cells were obtained from 21 of 23 cases, and the cells had the continuous passage and steady proliferation. Immunofluorescence staining showed that the cells positively expressed CK17 and CD44. It is indicated that primary cervical carcinoma cells can be obtained by trypsin and collagenase type Ⅰdigestion.   Related Articles | Metrics

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A new serum-free medium for hepatocytes growth Zhao Yi-chao1, Wang Yan1, Lu Hai1, Jian Guo-deng2, Pan Ming-xin1, Gao Yi1 2012, 16 (11):  1963-1968.  doi: 10.3969/j.issn.1673-8225.2012.11.015 Abstract ( 395 )   PDF (384KB) ( 335 )   Save BACKGROUND: A medium both for cell growth and avoiding adding serum medium is imminent, so serum-free medium emerge as the times require. OBJECTIVE: To develop a new serum-free medium for C3A cells to lay the foundation for the clinical application of hepatocytes and bioartificial liver. METHODS: C3A cells were cultivated with serum-free medium, complete medium and basal medium, respectively. The growth and morphological characteristics of cells were observed. Curves of cell growth and the energy were drawn. The leakage volume of alaninetransaminase, urea level and the level of albumin were determined by automatic biochemistry analyzer and radioimmunoassay. The changes of gene expression were determined by real-time fluorescence quantitative PCR. RESULTS AND CONCLUSION: The cells in the three kinds of mediums grew well. Apart from the 7th day, the cell density had no significant difference between the serum-free medium and complete medium groups in the culture period (P > 0.05), whereas it was obviously higher in these two groups than in the basal medium group at 4-7 days (P > 0.05). The level of urea in the serum-free medium group was significantly higher than that in the other two groups, and the level of urea in the complete group was higher than that in the basal medium group (P < 0.05). Real-time fluorescence quantitative RCR revealed that gene expressions of CK18, CK19 and hepatocyte nuclear factor 4α were lower in the serum-free group as compared with the complete medium group (P < 0.05). The results showed that the new serum-free medium for hepatocytes is effective just like the conventional complete medium.   Related Articles | Metrics

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Emergent isolation of myocardial cells from adult rats Wei Li-lan, Mo Shu-rong 2012, 16 (11):  1969-1972.  doi: 10.3969/j.issn.1673-8225.2012.11.016 Abstract ( 396 )   PDF (326KB) ( 510 )   Save BACKGROUND: To obtain high survival rate and good activity of the normal single myocardial cells is the basis of myocardial electrophysiology. It is important to find an experimental method which is close to the normal myocardial cells in morphology, function and structure. OBJECTIVE: To discuss the influence factors during emergent isolation of myocardial cells by using enzymatic method in rats and to build an emergent isolation method for stable obtaining myocardial cells to provide cells for myocardial electrophysiology and molecular biology. METHODS: The heart of Sprague-Dawley adult rats was rapidly excised after cervical dislocated and mounted on a Langendorff apparatus for aortic counter-current perfusion. It was perfused retrograde via the aorta for 5 minutes with calcium-free tyrode, and then perfused with collagenase dissolved in nominally calcium-free tyrode for 20 minutes. Ultimately calcium solution was perfused for 3 minutes. Then the rat ventricular cells were stored in HK solution. RESULTS AND CONCLUSION: Viability of freshly isolated myocardial cells which were rod-shaped with clear-striation was 60%-80%. It is shown that to adjust and control all the factors in the emergent isolation can ensure the quantity and quality of myocardial cells. Related Articles | Metrics

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PCR amplication of the DNA extracted from paraffin-embedded tissues in different conditions Ma Li-li, Ailijiang•Tuerxun, Zhang Li 2012, 16 (11):  1973-1976.  doi: 10.3969/j.issn.1673-8225.2012.11.017 Abstract ( 350 )   PDF (380KB) ( 479 )   Save BACKGROUND: Because of the damage of formalin-fixed paraffin-embedded tissue to DNA in the process of its production and preservation, it is difficult to extract high quality and sufficient DNA for PCR. OBJECTIVE: To analyze the influencing factors of the DNA extracted from paraffin-embedded tissue for PCR. METHODS: Genomic DNA was extracted from paraffin-embedded esophageal carcinoma tissue sections of 10.0 μm×2,      10.0 μm×4 and 10.0 μm×5. Template DNA amount of 0.05, 0.1, 0.2 μg was used to amplify gene β-actin, respectively. PCR cycle times were set as 35, 40 and 45. The influencing factors of PCR were analyzed. RESULTS AND CONCLUSION: The analysis of agarose gel electrophoresis showed that when paraffin-embedded esophageal carcinoma tissue section amount was 10.0 μm×2, the PCR cycle times were 40; when the template DNA amount was 0.05μg, the obtained target DNA was of the highest quality. It shows that reducing the amount of paraffin-embedded tissue and DNA template can be help to require high quality PCR strips; PCR cycle times should be more than 40 and less than 45, and if it increases beyond this range there is meaningless. Related Articles | Metrics

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Construction of an eukaryotic expression vector carrying amino acids 936-1132 of human telomerase reverse transcriptase and its expression in 293T cells Lu Jun-xiong, Jiang Hui-xian, Cao Ying, Pang Jian-xin 2012, 16 (11):  1977-1980.  doi: 10.3969/j.issn.1673-8225.2012.11.018 Abstract ( 374 )   PDF (426KB) ( 493 )   Save BACKGROUND: Human telomerase reverse transcriptase is an important component of telomerase. The display on telomerase activity results in anti-telomere shortening or cell senescence. OBJECTIVE: To construct an eukaryotic expressing vector containing the C-terminal amino acid 936-1132 of human telomerase reverse transcriptase; to express the constructed vector in human embryonic kidney 293T cells; to lay the foundation for the research of intracellular localization. METHODS: The C-terminal fragment of human telomerase reverse transcriptase containing amino acids 936-1132 and enzyme sites was amplified by PCR using human telomerase reverse transcriptase as templates. The amplified fragments were inserted into the eukaryotic expressing vector pcDNA3.1-HisA to construct recombinant plasmid pcDNA3.1-HisA-hTERT aa936-1132. This recombinant plasmid was transfected into the human embryonic kidney 293T cells, and then the protein expression was examined by western blot. RESULTS AND CONCLUSION: Double enzyme digestion and gene sequencing results showed that the recombinant plasmid pcDNA3.1-HisA-hTERT aa936-1132 containing the C-terminal amino acids 936-1132 of human telomerase reverse transcriptase was constructed successfully. The fusion protein was identified using western blot. Related Articles | Metrics

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Gene-silencing effect and small hairpin RNA vector construction of targeting human CCAAT/enhancer-binding protein-homologous protein gene You Yan-wen1, Xu Yu-ying1, Zhang Qin-xian2 2012, 16 (11):  1981-1984.  doi: 10.3969/j.issn.1673-8225.2012.11.019 Abstract ( 453 )   PDF (396KB) ( 395 )   Save BACKGROUND: Studies have proved that CCAAT/enhancer-binding protein-homologous protein (CHOP) is one of the three pathways of endoplasmic reticulum stress mediated apoptosis. There are few studies in home and abroad on endoplasmic reticulum stress mediated apoptosis in gastric carcinoma. OBJECTIVE: To construct an expression vector of a small hairpin RNA (shRNA) targeting human CHOP gene and to observe gene-silencing effects of CHOP in human gastric carcinoma cell BGC823. METHODS: The shRNA sequences targeting CHOP gene were designed and synthesized with two complementary oligonucleotide strands. The oligonucleotide strands were annealed and recombined into pSUPER-EGFP1 vector, which was identified by sequencing following transformation and amplification. The shRNA expression vector pSUPER-EGFP1-CHOP was transfected into human gastric carcinoma cell BGC823 via liposome. Reverse transcription-PCR and Western blot were used to detect expression levels of CHOP mRNA and protein in the transfected BGC823 cells, respectively. RESULTS AND CONCLUSION: The double-stranded oligonucleotide fragments of the shRNA targeting CHOP gene were cloned into pSUPER-EGFP1 vector and validated by sequence analysis which showed that expression vector pSUPER-EGFP1- CHOP was successfully constructed. Reverse transcription-PCR and Western blot indicated that CHOP mRNA and protein expressions were significantly decreased in the transfected cells, especially in those transfected with the shRNA targeting the sequence of CHOP-1, which induced 67% silencing of CHOP expression. Related Articles | Metrics

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Expression of interleukin-17 in the myocardium of rats with myocardial infarction Chen Zhao-zhe1, 2, Chen Zhi-jian1, Zeng Qiu-tang1, Liu Xue-gang1, Zhang Qing1, Liu Li-xin1, Zhu Yun-tao1, Qian Cheng1, Du Yi-mei1, Zhao Ning1 2012, 16 (11):  1985-1988.  doi: 10.3969/j.issn.1673-8225.2012.11.020 Abstract ( 239 )   PDF (406KB) ( 337 )   Save BACKGROUND: Expression of interleukin-17 increases in many cardiovascular disease such as autoimmune myocarditis, viral myocarditis, dilated cardiomyopathy and unstable coronary artery disease. Interleukin-17 plays an important role in chronic inflammatory process of cardiovascular diseases. OBJECTIVE: To study the expression of interleukin-17 in the myocardium after myocardial infarction in rats. METHODS: Liquid nitrogen freezing method was used to establish a rat myocardial infarction model. Totally 66 rats with modeling successful were randomly divided into six groups according to different observation times: 1, 3, 7, 14, 28, 56 days groups. And eight rats were selected as controls. The expression of interleukin-17 was examined by enzyme linked immunosorbent assay, laser confocal scanning microscope and Western blot. RESULTS AND CONCLUSION: Modeling success rate was 73%. With the development of myocardial infarction, interleukin-17 was abundantly expressed in the myocardial tissue and gradually increased over time. The expression of interleukin-17 in the 28 days group increased most significantly. Interleukin-17 as a potent pro-inflammatory factor can strongly express in rat myocardium after myocardial infarction, which may be related to the incidence of ventricular remodeling and arrhythmia. Related Articles | Metrics

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Construction of a rabbit ear model of chronic wound using transfixion ligation combined punch method Li Li1, Tong Ya-lin2, Zhan Qiu2, Lin Yuan1 2012, 16 (11):  1989-1993.  doi: 10.3969/j.issn.1673-8225.2012.11.021 Abstract ( 373 )   PDF (550KB) ( 335 )   Save BACKGROUND: In the past, the constructed rabbit ear models of chronic wound are limited to simple models of ischemia, while the model of both ischemia and congestion has been rarely reported. OBJECTIVE: To establish a chronic wound model for ischemia and congestion using rabbit ears. METHODS: A total of 40 rabbit ears were divided into two groups, experiment group and control group. The area of the dorsoventral side in ear root, all cephalic and center arteries and veins were transfixion ligated in the experiment group. Three circular wounds as deep as the cartilage surface were punched at the dorsal sides of the ears, using biopsy punch on the 10th day after ligation. Three circular wounds identical to that of the experiment group were punched in the control group. RESULTS AND CONCLUSION: The color of rabbit ears in the experiment group was deep violet-black immediately after ligation, and was somewhat violet-black, violet and lilac on the 10th, 20th and 30th days after ligation. It illustrates that the status of ischemia and congestion is durable and stable in the chronic wound model constructed using transfixion ligation combined punch method. Compared with the control group, the time of granulation tissue growth, the time of wound healing and epidermidalization in the experiment group were obviously delayed (P < 0.01). The expression of vascular endothelial growth factor in the experiment group was obviously lower than that of the control group (P < 0.01).These findings demonstrate that rabbit ear models of chronic wound constructed using transfixion ligation combined punch method are simple and reliable. It can be used as an ideal animal study model in the research of chronic wounds repair caused by ischemia and congestion factors. Related Articles | Metrics

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Development of secondary cardiovascular disease in hyperuricemia model rats induced by yeast extract combined with oteracil potassium Dilidaer•Xilifu1, Zhao Ping1, Song Li-juan1, Nijiati•Rehemu2, Zhang Xiang-yang1 2012, 16 (11):  1994-1998.  doi: 10.3969/j.issn.1673-8225.2012.11.022 Abstract ( 491 )   PDF (358KB) ( 811 )   Save BACKGROUND: The construction of hyperuricemia animal models is an important tool to study the relationship between hyperuricemia, renal disease and cardiovascular disease. OBJECTIVE: To establish hyperuricemia rat model using yeast extract combined with different doses of oteracil potassium and to explore the optimal dose for model establishment. METHODS: Rats were continuously treated with yeast extract or yeast extract combined with various doses of oteracil potassium by intragastric feeding or intraperitoneal injection for 28 days to establish rat hyperuricemia model. RESULTS AND CONCLUSION: Compared with the control group, there was significant increase in serum levels of uric acid in the groups of intragastric feeding with yeast extract alone, yeast extract combined with 50-200 mg/kg oteracil potassium or intraperitoneal injection with yeast extract combined with 50-100 mg/kg oteracil potassium; however, no remarkable effect on rat kidney, heart or artery was found. A continuously high level of serum uric acid and morphological and pathological changes in rat kidney, heart and arteries were detected when the dosage of oteracil potassium intraperitoneally injected combined with yeast extract was maximized to 200 mg/kg. These findings indicate that intraperitoneal injection of yeast extract combined with      200 mg/kg oteracil potassium is an optimal dosage for the construction of a persistent and stable hyperuricemia animal model; it is a favorable model for the study of pathological damages related to secondary cardiovascular diseases caused by hyperuricemia. Related Articles | Metrics

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Establishment of an inferior vene cava animal model by VX2 tumor implanted into the suprarenal and infrarenal vein in rabbits Zhang Yuan-guo, Ren Wei 2012, 16 (11):  1999-2002.  doi: 10.3969/j.issn.1673-8225.2012.11.023 Abstract ( 400 )   PDF (231KB) ( 330 )   Save BACKGROUND: At present, the principle and procedure for the treatment of tumor invading suprarenal and infrarenal inferior vene cave (IVC) are in dispute, but corresponding animal model is rarely reported in the literatures. OBJECTIVE: To establish the experimental animal model of the VX2 tumor invading suprarenal and infrarenal IVC, and to observe the influence on tumor growth and the IVC and abdominal aorta (AA). METHODS: Twenty-four rabbits were randomly divided into suprarenal group and infrarenal group, the prepared VX2 tumor tissue mass was transplanted into the soft tissues around the suprarenal and infrarenal IVC. Tumor size, diameter of the IVC and the AA, and the surrounding tissues were detected by ultrasound once a week as long as 5 weeks after implanted. RESULTS AND CONCLUSION: All VX2 tumors had been growing successfully in the experimental rabbits. Tumor size was significantly positive related to the growth time in two groups (r1=0.894, r2=0.879); the diameter of the IVC and AA were significantly negative related to tumor size in two groups (r3=-0.663, r4=-0.834; r5=-0.826, r6=-0.870). The difference of tumor size was not significant between two groups (P1=0.293>0.05). In the observation period, the stenosis of IVC was less than 50% in suprarenal group; the IVC in infrarenal group was completely blocked and effectively avoided the pressure on the AA. VX2 tumor tissue mass transplanted into the soft tissues around the infrarenal IVC leads to an ideal animal model.   Related Articles | Metrics

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Young rat model versus old rat model for cerebral ischemia Liu Zong-tao1, Qin Yong-sheng1, Liu Jiang2, Liu Yuan-xin3, Jiang Tao4 2012, 16 (11):  2003-2006.  doi: 10.3969/j.issn.1673-8225.2012.11.024 Abstract ( 263 )   PDF (226KB) ( 351 )   Save BACKGROUND: At present, rats are the most commonly used experimental animals in cerebral ischemia research. Rats have many advantages, such as inexpensive, good intraspecific homozygosity and similar cerebrovascular anatomical characteristics with human beings. OBJECTIVE: To explore the influence of aging to cerebral ischemia by observing the behavior changes in young and old rats with middle cerebral artery occlusion. METHODS: Young and old Sprague Dawley rats were randomly divided into four groups: young sham-operation group, old sham-operation group, young model group and old model group. Middle cerebral artery occlusion model was established by permanent ligation of the ipsilateral common carotid artery using the intraluminal suture technique. RESULTS AND CONCLUSION: The body mass index of the old sham-operation group was significantly lower than that of the young sham-operation group (P < 0.05), but higher than that of the old model group (P < 0.05). The body mass index of the old model group was lower than that of the young model group (P < 0.05). The modified neurological severity score of the old model group was significantly higher than that of the young model group on the 1st, 3rd, 5th and 7th days after operation (P < 0.05). The escape latency of the old model group was significantly longer than those of the other groups at the 3rd, 8th and 12th weeks after operation. The number of crossing platform location in the old model group was significantly less than those in the other groups   (P < 0.05, respectively). These findings demonstrated that compared with the young rat cerebral ischemia model with the same ischemia state, the old rat cerebral ischemia model showed more severe ischemia damage, lower repair capacity, and the neural functional recovery, spatial learning and memory ability was obvious inferior to that of the young rat model. It indicates that aging is one of the important factors that influence the neural injury after cerebral ischemia. Related Articles | Metrics

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Effects of Phyllanthus Emblica extracts on the insulin signal pathway in muscle and fat tissues of diabetic rats   Hu Wei 2012, 16 (11):  2007-2010.  doi: 10.3969/j.issn.1673-8225.2012.11.025 Abstract ( 321 )   PDF (309KB) ( 438 )   Save BACKGROUND: Phyllanthus Emblica has a significant hypoglycemic effect. However, its mechanism is not clear. OBJECTIVE: To explore the effects of Phyllanthus Emblica extracts on the proteins related to the insulin signal pathway in rat bone and fat tissues. METHODS: A total of 30 SD rats were randomly divided into control group, model group and Phyllanthus Emblica group. Rats in the latter two groups were intraperitoneally injected with streptozotocin to construct diabetic model. Rats in the Phyllanthus Emblica group were administrated with Phyllanthus Emblica extracts by gavage for 6 weeks; rats in the model group and the control group were administrated with distilled water of the same volume by gavage. RESULTS AND CONCLUSION: Compared with the control group, the body mass, fasting blood glucose, fasting insulin and insulin resistance index in diabetic rats increased significantly (P < 0.05); while the mRNA and protein levels of phosphatidylinositol-3-kinase, protein kinase B and glucose transporter 4 in the fat and muscle tissues decreased significantly     (P < 0.05, P < 0.01). The body mass, fasting blood glucose, fasting insulin and insulin resistance index in the Phyllanthus Emblica rats at 6 weeks after the gavage of Phyllanthus Emblica extracts was lower than those of the model rats; while the mRNA levels of phosphatidylinositol-3-kinase, protein kinase B and glucose transporter 4 in fat and muscle tissues increased significantly      (P < 0.01, P < 0.05). The protein expression of glucose transporter 4 in the fat tissues increased (P < 0.01). However, there was no significant difference between the protein expressions of glucose transporter 4 in the muscles. These findings indicate that Phyllanthus Emblica extracts can achieve hypoglycemic effects by regulating insulin-mediated phosphatidylinositol-3- kinase/protein kinase B/glucose transporter protein 4 signal transduction pathway. Related Articles | Metrics

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Effect of dietary iron contents on hematological iron status in male rats with long-term aerobic exercise Wang Jing, Kuang Gui-ran, Huang Hai-peng, Wang Xiao-yan, Yuan Li, Xiao De-sheng 2012, 16 (11):  2011-2014.  doi: 10.3969/j.issn.1673-8225.2012.11.026 Abstract ( 328 )   PDF (282KB) ( 430 )   Save BACKGROUND: Effects of exercise on hematological iron status have been extensively studied in female rats, but those effects have not been observed in male rats. OBJECTIVE: To investigate the effects of dietary iron contents and swimming exercise on hematological iron status of male rats. METHODS: Ninety weaning male rats were assigned into three groups: dietary iron deficiency group, dietary iron sufficiency group and dietary high iron content group, and then each group was further divided into exercise group and sedentary group. After one month of feeding with different iron contents diet, the rats in each exercise group entered a swimming program once a day, lasting for 3 months, and the rats in the corresponding sedentary groups received approximately the same amount of handling as the exercised rats, except for swimming. Animals were fasted for 24 hours after the last exercise regimen and then blood collection was taken from their hearts under pentobarbital anesthesia for analyzing related index of red blood cells and total iron binding capacity (TIBC). RESULTS AND CONCLUSION: Dietary iron contents could significantly affect the main effects of index of red blood and TIBC, and exercise could significantly affect the main effect of the red cell distribution and TIBC. In the dietary low iron content groups, the sedentary rats had the changes of iron deficiency with anemia, but the exercised rats had the significantly lower serum iron and transferrin saturation and the significantly higher TIBC. It showed that exercise under the condition of dietary iron deficiency could deteriorate serum iron status. Both in the dietary sufficient iron group and the dietary high iron content group, exercise could significantly increase red cell distribution and TIBC with no significant changes of other red blood cell indexes and serum iron, it suggested that exercise under the condition of dietary sufficient iron content may not induce the hematological low iron status.   Related Articles | Metrics

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Quality differences of bacterial genome DNA extracted from the human intestine by using QIAamp and Biomed DNA Stool Mini Kits Song Mei-ru1, Yao Ping1, Zhang Yue-xin2 2012, 16 (11):  2015-2018.  doi: 10.3969/j.issn.1673-8225.2012.11.027 Abstract ( 327 )   PDF (365KB) ( 765 )   Save BACKGROUND: Several studies have showed that there are different qualities of bacterial genome DNA extracted from the human intestine using different kits. Therefore, it is essential and urgent to select an effective, simple, and excellent DNA stool mini kit.　　　　　    OBJECTIVE: To compare the quality differences of bacterial genome DNA extracted from the human intestinal by using QIAamp and Biomed DNA Stool Mini Kits. METHODS: Thirty fresh fecal samples of healthy adult people were selected, and bacterial genome DNA was extracted using these two kits. Concentration, absorbance radio of 260 to 280 nm and extraction rate were measured. Species-specific primers for Lactobacillus group were designed by a set of 16S rDNA-targeted to conduct the general PCR using the genome DNA as the template. The electrophoresis strip’s number, brightness and density were compared after gel electrophoresis; subsequently, the number of Lactobacillus group was detected by fluorescent real-time PCR. RESULTS AND CONCLUSION: Concentration, extraction rate, expression level , Lactobacillus amount of bacterial genome DNA extracted from the human intestinal using QIAamp kit were higher than those using Biomed kit (P < 0.05 or P < 0.01). It suggests that the quality of genome DNA extracted with QIAamp kit is more excellent than that with Biomed kit.   Related Articles | Metrics

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Differentiation of monolayer cultured P19 embryonal carcinoma cells into myocardial cells induced by 5-azacytidine in vitro Wang Hai-ying 2012, 16 (11):  2019-2022.  doi: 10.3969/j.issn.1673-8225.2012.11.028 Abstract ( 308 )   PDF (489KB) ( 274 )   Save BACKGROUND: Several studies have shown that, during confluent culture and monolayer culture, P19 embryonal carcinoma cells can differentiate into myocardial cells induced by 5-azacytidine. OBJECTIVE: To observe whether P19 embryonal carcinoma cells can differentiate into myocardial cells induced by 5-azacytidine in vitro, as well as the expression of cardiac structural proteins. METHODS: P19 embryonal carcinoma cells were divided into two groups: experimental group induced by 1 μmol/L 5-azacytidine for 4 days, and control group induced by phosphate buffered saline. Double immunofluorescence staining technique was used to detect the co-expression of Desmin and cardiac troponin T. Western Blot was used to detect the expression of cardiac troponin T. RESULTS AND CONCLUSION: Co-expression of Desmin and cardiac troponin T was detected in the same part of the same cells in the experimental group, while no expression and co-expression of the two proteins in the control group. And the cardiac troponin T could be detected in the experimental group, while no expression in the control group. The P19 embryonal carcinoma cells under monolayer culture conditions could differentiate into myocardial cells, and express cardiac structure proteins after 5-azacytidine induction in vitro. Related Articles | Metrics

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Hydrogen metabolism in rabbits by different ways of hydrogen supply  Huang Guo-qing, Zhan Wei, Xiong Yan, Wen Ming-xiang, Li Ying-qing, Liao Xiao-xing 2012, 16 (11):  2023-2027.  doi: 10.3969/j.issn.1673-8225.2012.11.029 Abstract ( 299 )   PDF (307KB) ( 479 )   Save BACKGROUND: Different ways to supply hydrogen have curative effect on ischemia-reperfusion injury. But the relevant metabolism studies are rare to compare the different ways of supplying hydrogen. OBJECTIVE: To investigate the effect of intraperitoneal or intravenous saturated hydrogen saline and intraperitoneal injection of hydrogen on hydrogen metabolism in rabbits, providing a more optimal hydrogen treatment proposal. METHODS: Twenty New Zealand white rabbits were tested the basis concentration of exhaled hydrogen. Then all of them were supplied saturated hydrogen saline or hydrogen gas 10 mL/kg via intraperitoneal injection and saturated hydrogen saline       10 mL/kg via intravenous injection within 10 minutes, respectively. Exhaled hydrogen concentrations were monitored continuously. When the exhaled concentration declined to the baseline and stabilized for more than 30 minutes, hydrogen was supplied by another way. Time to maximal exhaled hydrogen concentration and max exhaled hydrogen concentration were recorded, as well as half-life and area under the curve were calculated to compare the effects of three different ways of hydrogen supply on hydrogen metabolism in rabbits. RESULTS AND CONCLUSION: The time to maximal exhaled hydrogen concentration was (3.25±1.80), (17.8±6.48) and (7.45±1.39) minutes in three ways, respectively, and there were significant differences among the three ways (P < 0.01). The half-life was (7.40±2.09), (141.50±85.01) and (1.20±0.37) minutes, respectively, and there were significant differences among the three ways (P < 0.01). The increment in the area under the curve was (302.17±221.90), (5 234.29±2 681.18), (209.51±     104.49) ppm•min, respectively, and there were significant differences among the three ways (P < 0.01). Hydrogen supply by intraperitoneal injection of saturated hydrogen saline or pure hydrogen both can rapidly increase exhaled hydrogen concentration in rabbits. Intraperitoneal hydrogen can maintain a high exhaled hydrogen concentration for a long time. Related Articles | Metrics

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Microdialysis and high performance liquid chromatography detection for vancomycin concentration in vitreous chamber of conscious rabbits★ Wang Ya-ling1, Wang Hong-ge1, Chen Xiao-long2 2012, 16 (11):  2028-2032.  doi: 10.3969/j.issn.1673-8225.2012.11.030 Abstract ( 287 )   PDF (268KB) ( 277 )   Save BACKGROUND: There are currently few studies regarding the pharmacokinetics of vancomycin via intravitreous injection. OBJECTIVE: To determine the concentration of vancomycin injected into the vitreous chamber of conscious rabbits.  METHODS: A microdialysis probe was implanted into vitreous chamber of normal rabbit eyes and rabbit eyes infected with bacterial endophthalmitis for 24 hours, and 10 g/L vancomycin 0.1 mL was administered intravitreally. The drug concentration in the vitreous chamber of rabbit eyes was determined at 0.5, 1, 2, 4, 6, 12, 24, 48, 72 and 84 hours after injection, through the microdialysis and high performance liquid chromatogram-ultraviolet detection. RESULTS AND CONCLUSION: The metabolism of vancomycin showed an open two-compartment model in normal rabbit eyes. Its half-life was 51.66 hours and the peak concentration was 695.92 mg/L. The metabolism of vancomycin in the infected vitreous chamber showed a one-compartment model. Its half-life was 11.91 hours and the peak concentration was 713.35 mg/L. All rabbits were injected with drugs for 84 hours and the intravitreous concentration of vancomycin was higher than minimal inhibitory concentration. The experimental findings indicate that microdialysis coupled to high performance liquid chromatography is a powerful tool to investigate the ocular pharmacokinetics of vancomycin, and the samples are harvested in a real-time, continuous and dynamic fashion when the experimental animals are conscious. Related Articles | Metrics

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Effects of oxidative stress and Rac1/2 on venous wall and their roles in traumatic deep vein thrombosis******☆ Li Xing-guo1, Zheng Hong-yu1, Li Wen2, Li Hong-kun1, Zhao Xue-ling1, Wang Bing1, 2012, 16 (11):  2033-2038.  doi: R318 Abstract ( 323 )   PDF (276KB) ( 375 )   Save BACKGROUND: The molecular mechanism and core regulatory network of deep vein thrombosis are not fully clarified yet. OBJECTIVE: To explore the roles of oxidative stress and Rac1/2 in rat deep vein thrombosis. METHODS: Deep vein thrombosis model in SD rats was established by a champing method femoral veins clamping combined with fixation of the lower extremity with plaster. The incidence and serious degree of thrombus were observed by dissecting rat femoral vein at different time points (2.5 and 25 hours after modeling). The model rats were divided into pre-thrombogenesis group (2.5 hours after modeling), thrombogenesis group (25 hours after modeling) and non-thrombogenesis group (25 hours after modeling). Then total RNA and protein were extracted from the femoral venous wall tissues. RESULTS AND CONCLUSION: Colorimetry results showed that compared with the non-thrombogenesis group, the concentration of malondiadehyde in rat femoral vein wall tissues of the thrombogenesis group was the highest (P < 0.05), followed by that of the pre-thrombogenesis group (P < 0.05). The concentrations of total superoxide dismutase and glutathione reductase in the thrombogenesis group were the lowest, followed by those in the pre-thrombogenesis group (P < 0.05). The results of gene chip hybridization analysis and real-time PCR showed that compared with the non-thrombogenesis group, the expressions of Rac1 and Rac2 in rat femoral vein wall tissues of thrombogenesis group increased the most, followed by that of the pre-thrombogenesis group (P < 0.05). These findings indicate that the up-regulation of malondialdehyde and Rac1/2 as well as the activity decrease of total superoxide dismutase and glutathione reductase may lead to the formation of deep venous thrombosis. Related Articles | Metrics

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Application of bone morphogenetic proteins in mandibular fast distraction osteogenesis Liu Hao, Wang Min 2012, 16 (11):  2039-2042.  doi: 10.3969/j.issn.1673-8225.2012.11.032 Abstract ( 262 )   PDF (358KB) ( 277 )   Save BACKGROUND: Distraction osteogenesis has became an effective method in treatment of different kinds of craniofacial deformities and bone defects. However, the major disadvantage of this method is the long distraction and consolidation period, which may lead to severe complications during the distraction process. OBJECTIVE: To summarize the research progress of the application of bone morphogenetic proteins (BMPs) in fast distraction osteogenesis. METHODS: A computer-based online search in Pubmed database was performed to select the reviews and papers related to BMPs and distraction osteogenesis from 1989 to 2011. RESULTS AND CONCLUSION: A total of 32 literatures on the the application of BMPs in fast distraction osteogenesis were included. BMPs have a strong osteogenic activity and capability of promoting regeneration and remodeling of bone. The present research shows that BMPs can be used to accelerate new bone formation and shorten the treatment period. However, the clinical application of BMPs needs further research. Related Articles | Metrics

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Osteoblasts culture in bone tissue engineering and exercise research  Li Li-yan, Ren Tang-ke 2012, 16 (11):  2043-2046.  doi: 10.3969/j.issn.1673-8225.2012.11.033 Abstract ( 266 )   PDF (422KB) ( 419 )   Save BACKGROUND: Under different culture and cytokine-stimulated conditions, steoblasts have different proliferation and differentiation abilities as well as functions.  OBJECTIVE: To summarize the progress in in vitro culture of osteoblasts as well as proliferative ability of osteoblasts. METHODS: A search of PubMed and Wanfang databases (1990/2011) was performed for articles addressing exercise, osteoblast culture and bone tissue engineering. The keywords were “exercise, osteoblast raise method” in English and “exercise, osteoblast, bone tissue engineering” in Chinese. Unrelated articles or repetitive articles were excluded, and finally 29 articles were included in result analysis. RESULTS AND CONCLUSION: Osteoblast culture is influenced by the following factors: methods to choose seed cells, pattern of scaffold materials, cytokines, culture environment, and mechanical factors in bone tissue engineering construction. Osteoblasts cultured in different conditions and stimulated by different cytokines have different proliferative and differentiated abilities. The proliferative speed of osteoblasts is closely related to a low shear environment produced by rotating-wall vessel bioreactor, appropriate cytokines and selection of cytoskeleton. Related Articles | Metrics

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Stress stimulation and signal pathways of osteoprotegerin, receptor activator of nuclear factor kappa B and its ligand in bone metabolism  Wang Dan, Bao Jie, Wang Guo-xiang 2012, 16 (11):  2047-2050.  doi: 10.3969/j.issn.1673-8225.2012.11.034 Abstract ( 236 )   PDF (432KB) ( 426 )   Save   BACKGROUND: Signal pathway of osteoprotegerin, receptor activator of nuclear factor kappa B and its ligand in bone metabolism is one of the accesses to cope with the stress sensitivity. Different types of the exercise can produce different types of stress stimulation, and influence the signal pathways in bone metabolism. OBJECTIVE: To explore the influence of different types of exercise on the signal pathways of osteoprotegerin, receptor activator of nuclear factor kappa B and its ligand in bone metabolism. METHODS: An online search of CNKI, HighWire and Elsevier databases was performed for articles related to “stress stimulation, osteoprotegerin, receptor activator of nuclear factor kappa B and its ligand”. Articles published from 2000 to 2011 with the keywords of “stress stimulation, osteoprotegerin, receptor activator of nuclear factor kappa B and its ligand” or “stress stimulation, bone metabolism” were included. Articles related to the effects of stress stimulation on the signaling pathways were selected. Articles published recently or in the authorized journals were of priority. A total of 215 articles were collected in the initial research, and 31 of them were retained according to inclusion and exclusion criteria. RESULTS AND CONCLUSION: Exercise continuously produces stress stimulation on bone. This mechanical stress stimulation regulates the bone tissue metabolism by affecting the signal regulating system of osteoprotegerin, receptor activator of nuclear factor kappa B and its ligand. But the results in relevant articles are inconsistent. Therefore further study is needed.   Related Articles | Metrics

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Genetics development of human body muscle short-time service ability Wang Shi-jun 2012, 16 (11):  2051-2054.  doi: 10.3969/j.issn.1673-8225.2012.11.035 Abstract ( 304 )   PDF (404KB) ( 380 )   Save BACKGROUND: In recent years, researchers have demonstrated that, from the perspective of human genetics, human muscles short-term service ability is largely controlled by genes, and body muscle strength and anaerobic endurance have a higher heritability. OBJECTIVE: To review some findings between genetic factors and muscle short-time service ability in order to search new ways and ideas for the further research. METHODS: A computer-based search of CNKI, PubMed and Medline databases was performed for articles published between 1990 and 2011 regarding genetic studies of muscle short-time service ability. Finally, 35 papers were included in result analysis. RESULTS AND CONCLUSION: The article summarizes the early animal experiments and recent human trials. The results from the genetic research of human muscle short-time service ability achieved an amazing consistency and exhibited a high positive rate. Thus, individual differences existing in the human muscle short-time service ability reflects the DNA sequence and its differences in expression regulation to a large extent. Along with the development of molecular biology techniques, these studies provide a strong theoretical basis for further genetic research of gene-related muscle short-time service ability.   Related Articles | Metrics

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Exercise and apoptosis in different tissues Xu Su-yang, Wang Wen-di, Fu Yu, Su Quan-sheng 2012, 16 (11):  2055-2058.  doi: 10.3969/j.issn.1673-8225.2012.11.036 Abstract ( 211 )   PDF (379KB) ( 312 )   Save BACKGROUND: Apoptotic process can be activated by evoked movement and cause cell death. OBJECTIVE: To analyze the effects of exercise training on apoptosis in different tissues, to discuss the relationship of exercise fatigue, sports injuries, athletic heart remodeling, exercise-induced minor myocardial injury, exercise-induced immunosuppression and apoptosis in different tissues. METHODS: An online search of Medline database was performed using key word of “apoptosis” for articles published from January 1990 to May 2010. Articles on relations between exercise training and apoptosis were included. Meanwhile, another online search of Ginii database was performed using key words of “アポトーシス，運動” for relevant articles published from January 2000 to June 2010. And a third online search of China National Knowledge Infrastructure database was performed using key words of “apoptosis, exercise training, skeletal muscle, myocardium, lymphocytes, liver and kidney cells” in Chinese for relevant articles published from January 2000 to May 2010. RESULTS AND CONCLUSION: Single exercise and systematic exercise training both affect apoptosis in skeletal muscle, myocardium, lymphocytes, liver cells and kidney cells. Apoptosis level increases with the increasing of exercise intensity. These findings indicate that apoptosis is closely related to exercise-induced skeletal muscle micro-injury, athletic heart remodeling and exercise-induced minor myocardial injury. Related Articles | Metrics

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“TIME” principles and its clinical significance in wound repair  Xu Yuan, Liu Hong-wei 2012, 16 (11):  2059-2062.  doi: 10.3969/j.issn.1673-8225.2012.11.037 Abstract ( 559 )   PDF (381KB) ( 609 )   Save BACKGROUND: While the cellular and molecular mechanisms of wound healing have been understood gradually and the experiences of wound bed management have been accumulated, the importance of wound bed preparation to wound repair has recently attracted more attention. OBJECTIVE: To fully analyze the mechanisms of wound healing, various influence factors and their effects in this progress. METHODS: An on line search of CNKI and PubMed databases was performed for articles published between 1994 and 2011, using key words of “TIME principles, wound bed preparation, wound healing” in Chinese and English, respectively. The clinical application principles and outcomes of wound healing were analyzed. RESULTS AND CONCLUSION: A total of 498 papers were collected, and 24 of them were included according to inclusive criteria. TIME is the initials of four principles in wound bed management. T means to remove the necrotic tissue. I means to control the infection and to reduce the inflammation. M means to keep the wound moisture under normal condition for the granulation tissue growth and re-epithelialization. E means to remove non migrating epidermis at the edge of wound. As a valuable guidance tool for wound management, the TIME concept has been applied to clinical practice, and has been widely appreciated by clinicans.   Related Articles | Metrics

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Influence of exercise on mitochondrial biological characteristics and cell senescence  Fu Yu, Wang Wen-di, Xu Su-yang, Su Quan-sheng 2012, 16 (11):  2063-2066.  doi: 10.3969/j.issn.1673-8225.2012.11.038 Abstract ( 316 )   PDF (402KB) ( 892 )   Save BACKGROUND: Changes in mitochondrial structure and function are closely linked with cell senescence. OBJECTIVE: To sum up the influence of changes in mitochondrial biological characteristics on cell senescence. METHODS: A computer-based search of Medline database was performed for articles about the influence of mitochondria and exercise on cell senescence published from January 1990 to June 2011 and the related articles published from January 1994 to June 2011 were searched in China Journal Full-text Database and Wanfang database. RESULTS AND CONCLUSION: Reactive oxygen species originated from the mitochondrial respiratory chain lead to increasing mitochondrial membrane permeability, reducing mitochondrial transmembrane potential and decreasing adenosine-triphosphate synthesis. Sustained oxidation of mitochondrial reactive oxygen species increases the mitochondrial DNA damage, leads to serious damage in mitochondrial structure and function, and then promotes cellular senescence or even death. Aerobic training can significantly improve the body aerobic metabolism level, increase aerobic capacity, improve the activities of body mitochondrial respiratory chain enzyme and matrix enzyme, reduce the glycolytic activity, mitochondrial oxidative damage and the rate of mitochondrial DNA deletion mutation. Therefore the mitochondrial adaptation and oxidative phosphorylation activity are improved. In training, reasonable arrangements of intermittent exercise should be considered as well as avoiding the cell damage of continuing decline in mitochondrial structure and function caused by high intensity training. It may play a positive role in delaying the cell aging and organism aging. Related Articles | Metrics

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Application of chicken embryo in biomedical research Wang Yong, Li Li, Li Tao, Duan Yong-gang 2012, 16 (11):  2067-2070.  doi: 10.3969/j.issn.1673-8225.2012.11.039 Abstract ( 317 )   PDF (448KB) ( 789 )   Save BACKGROUND: In recent years, there are a lot of researches on tissue culture and cell culture using chicken embryo. But neurocyte culture using chicken embryo has not been reported. OBJECTIVE: To summarize the research progress of tissue culture and cell culture using chicken embryo in order to find a convenient and inexpensive method of neurocyte culture. METHODS: An online search of CNKI and PubMed databases was performed for articles related to chicken embryo culture model and neurocyte culture published between January 1998 and June 2011, using key words of “chicken embryo chorioallantoic membrane model; neuronal culture” in Chinese and English. Articles related to chicken embryo culture model were selected. As to the in identical field, articles published recently or in authoritative magazine were of priority. A total of 205 articles were collected in the initial search, 34 of them were retained according to the inclusion criteria. RESULTS AND CONCLUSlON: Chicken embryo is widely used in virus isolation, virus cultivation, vaccine preparation, mechanisms of tumor biology and drug screening. Compared with other culture methods, the chicken embryo culture model is cheap, convenient and rapid. Fully understanding and correctly utilization of the chicken embryo culture model provide scientific and feasible methods and train of thought for the research in many fields. Related Articles | Metrics

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Application of saliva test in sports-related biochemical indexes analysis Li Min-hua, Tang Jian 2012, 16 (11):  2071-2075.  doi: 10.3969/j.issn.1673-8225.2012.11.040 Abstract ( 384 )   PDF (469KB) ( 542 )   Save BACKGROUND: Significant correlations are observed between saliva and blood in evaluating biochemical parameters. Therefore, more and more researchers take saliva test as another important method to monitoring organism biochemical parameters. Besides, more and more researchers in sport and exercise science take saliva test as a convenient and effective method of long-term operation test. OBJECTIVE: To multianalysis of saliva test as a method of the individual biochemical parameters, including operation, advantages, disadvantages, and feasibility analysis. So this method can be an effective tool to test individual physical condition and mental state. METHODS: A computer-based online search of PubMed database (http://www.ncbi.nlm.nih.gov/pubmed/) and CNKI database (http://www.cnki.net/) was performed for papers published from 1980 to 2011 with the key words of “saliva, blood, test, indictor, sport, excercise”. Analysis and choosing those papers related to saliva test applied in evaluating biochemical parameters, especially applied in exercise. Published earlier, repeated and similar studies were excluded. RESULTS AND CONCLUSION: Thirty-one literatures were included and it can be concluded that saliva analysis is a convenient and effective tool for monitoring biochemical parameters. Significant correlations were reported between salivary and blood   concentrations including salivary immune-globulin A, cortisol, testosterone, dehydroepiandrosterone and amylase, and these biochemical parameters have been extensive used. As the development of the research, saliva analysis will be a significant tool for assessing individual physiological and psychological conditions with exercise and training. Related Articles | Metrics

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Gangliosides for acute cerebral infarction: A systematic review based on domestic randomized controlled trials Deng You-qi, Yang Xiao-hua 2012, 16 (11):  2076-2079.  doi: 10.3969/j.issn.1673-8225.2012.11.041 Abstract ( 295 )   PDF (307KB) ( 436 )   Save BACKGROUND: Some studies have reported that gangliosides are safe and effective in the treatment of acute cerebral infarction. However, ganglioside effects on cerebral infarction are still uncertain due to few samples of a single study. OBJECTIVE: To evaluate the efficacy and safety of gangliosides for acute cerebral infarction. METHODS: Literature search was carried out by computer and manual search. We searched for randomized controlled trials about gangliosides for acute cerebral infarction in CBMdisc (1990/2010), VIP (1990/2010), CNKI (1990/2010) and major neurological periodicals. Quality assessment was conducted according to the Cochrane Handbook for Systematic Reviews of Interventions 5.0.1. Meta-analysis was performed for the results of homogeneous studies using RevMan 5.0 software． RESULTS AND CONCLUSION: Totally 119 articles were retrieved, and finally nine randomized controlled trials were included, but the methodological quality was relatively low. Meta analysis showed that the effectiveness of gangliosides was superior to that of control treatment. Due to the low quality of included trials, the effect and safety of gangliosides in the treatment of acute cerebral infarction have to be verified by further high quality evidence.   Related Articles | Metrics

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Meta-analysis of the relationship between PstI/RsaI polymorphism of cytochrome P450 2E1 gene and colorectal cancer susceptibility  Feng Liang, Zhang Hui-rui, Lü Yan-dong, Yang Tuo-yun 2012, 16 (11):  2080-2083.  doi: 10.3969/j.issn.1673-8225.2012.11.042 Abstract ( 272 )   PDF (316KB) ( 375 )   Save BACKGROUND: Cytochrome P450 2E1 gene (CYP2E1) is one of the important encoding genes in its enzyme system. There are many studies on the relationship between CYP2E1 polymorphism and colorectal cancer susceptibility. However the results have some difference. OBJECTIVE: To explore the relationship between CYP2E1 PstI/RsaI single neucleotide polymorphism and colorectal cancer susceptibility. METHODS: Studies on the relationship between CYP2E1 PstI/RsaI single neucleotide polymorphism and colorectal cancer susceptibility were obtained by searching EMBASE database, PubMed database, Ovid database, Highwire press database, CNKI database, CBM database and Wanfang database. The odds ratio (OR) of CYP2E1 allele of the case group and control group was taken as effective index. Fixed or random effect Meta-analysis model was used to calculate the combined OR. RESULTS AND CONCLUSION: A total of 9 case-control studies containing 4 760 patients and 5 812 controls were included according to inclusion criteria. Evaluation of risk of rectal cancer in mutation homozygote when wild homozygote was used as reference: OR=1.24 (OR=1.24, 95% confidence interval (CI): 0.93-1.66, P=0.15); Evaluation of risk of rectal cancer in mutation homozygote when wild homozygote and mutation homozygote were used as reference: OR=1.26 (OR=0.94, 95%CI: 0.94-1.68, P=0.12); Evaluation of risk of rectal cancer in mutation homozygote and wild homozygote when wild homozygote was used as reference: OR=1.00 (OR=1.00, 95%CI: 0.90-1.12, P=0.97). Sub-group analysis: the studies were divided into Caucasian populations and East Asian populations; Only the Caucasian populations showed a relationship between CYP2E1 PstI/RsaI single neucleotide polymorphism and colorectal cancer susceptibility in the c2c2 VS c1c1 model (OR=2.67, 95%CI: 1.03-6.89, P=0.04). It is indicated that there is no close relationship between CYP2E1 PstI/RsaI single neucleotide polymorphism and colorectal cancer susceptibility except the Caucasian in c2c2 VS c1c1 model. Related Articles | Metrics

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Standing time and bone density changes in patients with spinal cord injury Li Qing-gui, Guo Zhao, Wang Xiao-dong 2012, 16 (11):  2084-2086.  doi: 10.3969/j.issn.1673-8225.2012.11.043 Abstract ( 337 )   PDF (284KB) ( 303 )   Save BACKGROUND: Osteoporosis after spinal cord injury can decline pragmatic ability and seriously influence life quality. OBJECTIVE: To analyze the activities and bone density changes in patients with spinal cord injury. METHODS: Patients with spinal cord injury were divided into three groups according to daily standing time. Patients in sufficient activity group with daily standing time of more than 1 hour, patients in restricted activity group with daily standing time of less than 1 hour, and non-activity group has no standing activities. RESULTS AND CONCLUSION: Dual-energy X-ray absorptiometry detected bone mineral density in lumbar spine and femur showed that bone densities of lumbar spine (L2-4), femoral neck, femoral intertrochanteric and Ward's triangle in the sufficient activity and restricted activity groups were higher than those of the non-activity group (P < 0.05), and bone density in the sufficient activity group was superior to the restricted activity group (P < 0.05). The results confirmed that compared with relative restricted activity after spinal cord injury, appropriate standing activities can help to reduce bone loss, alleviate the development of osteoporosis and improve the level of bone density. Related Articles | Metrics

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A novel 5’ splice site mutation in ACTC1 gene may play an important role in congenital ventricular septal defect★ Li Hang1, Wang Bin-bin2, Gao Bing-ren1, Liu Jiang-yan3, Zhao Qi-ming1, Chen Wen-sheng1, Wang Wei1, Yang Kun1 2012, 16 (11):  2087-2090.  doi: 10.3969/j.issn.1673-8225.2012.11.044 Abstract ( 430 )   PDF (276KB) ( 455 )   Save BACKGROUND: As a candidate gene of congenital heart disease, ACTC1 gene is related to congenital atrial septal defect in humans. OBJECTIVE: To perform a mutation screen of ACTC1 gene in 110 nuclear families of congenital heart disease. METHODS: A case-control study was conducted based on 110 nuclear families of congenital heart disease and 300 normal human beings with no reported cardiac malformation. Six fragments in the coding region of ACTC1 gene was amplified by PCR in vitro using five primers pairs. PCR products were screened for gene mutations. RESULTS AND CONCLUSION: A novel G-to-A variant was found at the third nucleotide of the intron downstream from exon 5. This mutation existed in a 5-year-old female with an isolated ventricular septal defect and her 30-year-old father, who had no reported cardiac anomalies. This mutation was not detected in 300 normal controls. These findings indicate that the mutation may be related with congenital ventricular septal defects in humans. Related Articles | Metrics