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10 December 2011, Volume 15 Issue 50 Previous Issue    Next Issue

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Relationship between synovium pathological change and injection timing in rabbits with papain-induced knee osteoarthritis Sun Lu-ning, Zhao Yan-hua, Huang Gui-cheng, Xia Jian-long 2011, 15 (50):  9311-9313.  doi: 10.3969/j.issn.1673-8225.2011.50.001 Abstract ( 338 )   PDF (788KB) ( 684 )   Save BACKGROUND: A stable degenerative knee osteroarthritis model in rabbits can be established by the induction of papain. OBJECTIVE: To explore the relationship between the pathological change of synovium and injection timing in rabbits with knee osteoarthritis induced by intraarticular injection of papain. METHODS: Papain solution was injected into the right knee joint cavity of 15 New Zealand white rabbits on the 1st, 4th and 7th days to induce knee osteoarthritis in rabbits . The general and histopathologic changes of knee synovium were examined in the 2nd, 4th and 6th weeks after first injection. Five rabbits were examined at each time point. RESULTS AND CONCLUSION: The synovium inflammatory reaction of rabbit knee joint was the most serious at the 2nd week after the first injection of papain, characterized by the most obvious proliferation of synovial cell, obvious proliferation of vascular tissue and substratum porosity fiber vascular tissue, visible infiltration of many lymphoplasmacytoid cells. The inflammatory reaction relieved gradually in the 4th and 6th weeks. These results illustrate that the synovium inflammatory reaction of knee joint in rabbits reach its maximum effect in the 2nd week after the first injection of papain. Related Articles | Metrics

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Safety and feasibility of ultrasound-targeted microbubble destruction applied for transfection of enhanced green fluorescent protein plasmid into the femoral head of rabbits Peng Hao, Huang Lei, Ding Shuai, Li Bin-bin, Gan Jing-yue, Xu Shun-en 2011, 15 (50):  9314-9318.  doi: 10.3969/j.issn.1673-8225.2011.50.002 Abstract ( 280 )   PDF (527KB) ( 435 )   Save BACKGROUND: In recent years, ultrasound microbubble gene transfer system has been applied for gene transfection in many parts of the body, but it has been seldom reported to be used for gene transfection in bone parts. OBJECTIVE: To investigate the efficiency and feasibility of ultrasound-targeted microbubble destruction applied for transfection of enhanced green fluorescent protein plasmid into the femoral head of rabbits.  METHODS: Japanese big-ear rabbits were randomly divided into five groups: bare transfection, pre-irradiation + bare transfection, ultrasound transfection, pre-irradiation+ultrasound transfection, and repeatable transfection. In the first two groups, ultrasound-targeted gene transfection and irradiation was not used, but in the latter three groups, ultrasound-targeted microbubble destruction was used to transfect enhanced green fluorescent protein (EGFP) plasmid into the femoral head of rabbits. At 1 week after transfection, EGFP expression in femoral head was observed under the fluorescence microscope. RESULTS AND CONCLUSION: EGFP expression appeared in the ultrasound transfection, pre-irradiation + ultrasound transfection and repeatable transfection. The transfection efficiency of EGFP plasmid was significantly higher in the repeatable transfection group than in the other groups (P < 0.01). Obvious injury loci were not observed in the soft tissue and bone tissue slices of ultrasonic irradiation parts in the ultrasound transfection, pre-irradiation + ultrasound transfection and repeatable transfection groups. These results confirm that ultrasound-targeted microbubble destruction is a safe and effective method to transfect EGFP plasmid into the femoral head of rabbits. Related Articles | Metrics

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Bone tissue function and pathological changes of hormone-induced avascular necrosis of the femoral head in rabbits Yang Wei-qiang, Jiang Zhen-gang, Huang Chang-lin 2011, 15 (50):  9319-9322.  doi: 10.3969/j.issn.1673-8225.2011.50.003 Abstract ( 276 )   PDF (1252KB) ( 489 )   Save BACKGROUND: So far, research on hormone-induced avascular necrosis of the femoral head does not involve cell function change induced by the alteration of osteoblast and osteoclast. OBJECTIVE: To investigate bone metabolism and bone tissue pathological changes of hormone-induced avascular necrosis of the femoral head in rabbits. METHODS: A number of 32 New Zealand white rabbits were randomly divided into two groups, control group and model group. The model rabbits were injected with 8.0 mg/kg hydrocortisone acetate twice a week for 8 weeks to produce avascular necrosis model of the femoral head. The control rabbits were injected with equivalent physiological saline. RESULTS AND CONCLUSION: Compared with the control group, the serum calcium and phosphate level of model group dropped significantly since the 2nd week (P < 0.05), while the enzymatic activity of tartrate-resistant acid phosphatase rised significantly (P < 0.05). Bone specific alkaline phosphatase level of model group increased while bone gla protein level decreased at the start of the 4th week (P < 0.05). Bone specific alkaline phosphatase level of model group declined by the 8th week. Compared with the contrl group, the femoral intertrochanter bone mineral density of model group dropped since the 2nd week (P < 0.05), and the bone mineral density of femoral head dropped since the 4th week (P < 0.05). From the 4th week, the amount of osteoblast decreased significantly, while the osteoclast amount increased significantly (P < 0.05). These results suggest that hormone can damage bone shaping and bone remodeling by damaging bone cells, reduce the content of mineral salt used for bone formation, and lead to hormone-induced avascular necrosis of the femoral head. Related Articles | Metrics

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Purified rat chondrocytes can be gained by using typeⅡ collagenase enzyme digestion method in short time Liu Zhen-feng, Fang Rui, Meng Qing-cai 2011, 15 (50):  9323-9326.  doi: 10.3969/j.issn.1673-8225.2011.50.004 Abstract ( 291 )   PDF (1377KB) ( 622 )   Save BACKGROUND: To establish an economic, efficient, practical isolation and culture system of chondrocytes has important significance to cartilage in vitro experiments. OBJECTIVE: To investigate and improve the culture method of rat articular chondrocytes. METHODS: Rat cartilage of bilateral hip and knee was resected from one week old male rats under aseptic condition. Chondrocytes were isolated by typeⅡ collagenase enzyme digestion method. The cells were cultured and passaged, then identified. RESULTS AND CONCLUSION: Inverted phase contrast microscope observation showed that the primary cultured chondrocytes adherented at the 12th hour after cultivation. The monolayer formation occurred on the 3rd day after cultivation. And cells were ready to be passaged on the 4th day after cultivation. After passaged for six generations, some cells represented a spindle-like appearance. In the 7th generation, most cells turned into a long and irregular appearance, and cell proliferation capacity diminished. Toluidine blue staining showed that the nucleis of cultured chondrocytes were metachromatic. Immunofluorescent staining showed that cultured chondrocytes had a positive expression of collagen type Ⅱ. These findings illustrate that a large number of purified rat chondrocytes can be gained by this method in a short time. Related Articles | Metrics

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Effects of Uygur medicine Maizhuni medicated serum on the expression of matrix metalloproteinase 1 and tissue inhibitor of metalloproteinase 1 in rat chondrocytes Fang Rui, Liu Zhen-feng, Ailijiang•Asila, Hong Han-gang, Deng Ying-jie, Meng Qing-cai 2011, 15 (50):  9327-9330.  doi: 10.3969/j.issn.1673-8225.2011.50.005 Abstract ( 278 )   PDF (1187KB) ( 486 )   Save BACKGROUND: Imbalance of matrix metalloproteinase 1 and its tissue inhibitor is the key of articular cartilage degradation in osteoarthritis. OBJECTIVE: To explore the effect of Uygur medicine Maizhuni on the expression of metalloproteinase 1 and tissue inhibitor of metalloproteinase 1 in rat articular cartilage cells induced by interleukin-1β. METHODS: Primary chondrocytes were isolated from one week old SD rats, and then identified. Chondrocytes of passage 2 were randomly assigned into control group, model group and Maizhuni group. Chondrocytes in model group and Maizhuni group were treated with 10 μg/L interleukin-1β for 24 hours. After that, chondrocytes in Maizhuni group and model group were further cultured for 12, 24 and 48 hours in addition with 10% Maizhuni medicated serum and rat serum, respectively. RESUILS AND CONCLUSION: According to RT-PCR results at 48 hours after cultivation, 10% Maizhuni medicated serum increased the mRNA expression of matrix metalloproteinase 1 tissue inhibitor and suppressed the mRNA expression of matrix metalloproteinase 1 in chondrocytes. But the effect was not obvious at 24 and 36 hours after cultivation. Immunofluorescence and cytochemical staining results showed that Maizhuni medicated serum increased the expression of matrix metalloproteinase 1 tissue inhibitor in chondrocytes. These findings indicate that Maizhuni can regulate the expression of metalloproteinase 1 and its tissue inhibitor in rat chondrocytes induced by interleukin-1β in a time-dependent manner. Related Articles | Metrics

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Culture of human bronchial epithelial cells in vitro Wu Qi-biao, Lu Jin-fu, Yin Yao-ting, Cao Shi-hong 2011, 15 (50):  9331-9334.  doi: 10.3969/j.issn.1673-8225.2011.50.006 Abstract ( 370 )   PDF (1411KB) ( 584 )   Save BACKGROUND: Culture of human bronchial epithelial cells has been increasingly applied in the research of respiratory diseases. OBJECTIVE: To explore a feasible in vitro culture method for human bronchial epithelial cell line supplied by American Type Culture Collection. METHODS: Different culture conditions of human bronchial epithelial cells were tested with trials and errors. A feasible method with DMEM/F12 containing 20% fetal bovine serum was finalized. RESULTS AND CONCLUSION: The cultured human bronchial epithelial cells were flat, polygonal, and displayed a cobblestone-like distribution when overgrown. Immunohistochemical staining results showed an expression of epithelial marker keratin in cultured cells. The expression of nuclear factor κB, tumor necrosis factor α and interleukin 8 was increased significantly in cultured cells exposed to sputum supernatant from patients with bronchiectasis. These findings demonstrated that the cultured human bronchial epithelial cells were obtained successfully. Therefore, there is no need to stick to the culture condition recommended by American Type Culture Collection. The modified culture method is simple, feasible and valuable. Related Articles | Metrics

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Cultivation of mouse mammary epithelial cells by using improved collagenase/trypsin digestion method Yi Qiong, Wang Lu, Li Yuan-fang, Zhu Wei 2011, 15 (50):  9335-9338.  doi: 10.3969/j.issn.1673-8225.2011.50.007 Abstract ( 364 )   PDF (1152KB) ( 462 )   Save BACKGROUND: The mixture of collagenase A and trypsin that used to isolate the mammary epithelial cells is complex and strict to operate. OBJECTIVE: To clarify whether mammary epithelial cells can be successively cultured with improved collagenase/trypsin digestion method in vitro. METHODS: The mixture of typeⅠand Ⅱcollagenase and trypsin (1:1:1) was used to digest breast tissue, which was shredded directly with ophthalmic scissors. The organoids were obtained by 37 ℃ oscillation extraction and the differential adhesion method was used to remove the fibroblasts. Then the organoids were inoculated in the plastic dish. RESULTS AND CONCLUSION: Inverted microscope observation showed that epithelial organoids could be mostly acquired and had attached to the plastic dish after 12 hours, and then cells were spreading out from them and forming confluent monolayer of cobblestone after cultured for 72 hours. In addition, the tissue-specific expression of cytokeratin 18 in mammary epithelial cells was identified by cytokeratin immunohistochemistry. It indicated that plenty of pure mammary epithelial cells could be harvested by using improved collagenase/trypsin digestion. Related Articles | Metrics

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Human umbilical vein endothelial cells damaged by cigarette smoke extract and intervention of atorvastatin Huang Xu, Dai Yuan-bin, Wu Tian-fang, Wang Li-wei, Tang Wei-xue 2011, 15 (50):  9339-9343.  doi: 10.3969/j.issn.1673-8225.2011.50.008 Abstract ( 328 )   PDF (1528KB) ( 516 )   Save BACKGROUND: Smoking is an important risk factor for many vascular ischemic diseases, such as arterial occlusive and thromboangiitis obliterans. However, the exact mechanism behind this is unclear. OBJECTIVE: To investigate the effects of cigarette smoke extract on human umbilical vein endothelial cells and the intervention effect of atorvastatin during this process. METHODS: Human umbilical vein endothelial cells were cultured in vitro and divided into three groups, control group, cigarette smoke extract group and cigarette smoke extract plus atorvastatin group. Control group were cultured in DMEM medium containing 10% fetal bovine serum. Cigarette smoke extract group were cultured in DMEM medium containing 10% fetal bovine serum and 10% cigarette smoke extract. Cigarette smoke extract plus atorvastatin group were cultured in DMEM medium containing 10% fetal bovine serum, 10% cigarette smoke extract and 10 μmol/L atorvastatin. RESULTS AND CONCLUSION: Compared with control group, the cell morphology in cigarette smoke extract group was much irregular, the survival rate, the nitric oxide level in cell supernatant and the enzyme activity of nitric oxide synthase was decreased (P < 0.05). The cell morphology in cigarette smoke extract plus atorvastatin group was improved, the survival rate, the nitric oxide level in cell supernatant and the enzyme activity of nitric oxide synthase was increased (P < 0.05). It indicates that atorvastatin can protect vein endothelial cells from the damage effects of cigarette smoke extract. The mechanism may be related to the endothelium cell growth, the enzyme activity of nitric oxide synthase and the release of nitric oxide. Related Articles | Metrics

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Interaction between primary cultured human thymocytes and human immunodeficiency virus Gao Nan-jun, Chen Ying-jie, Li Xue-tao, Kong Wei-yun, Zhou Yi-ping 2011, 15 (50):  9344-9347.  doi: 10.3969/j.issn.1673-8225.2011.50.009 Abstract ( 306 )   PDF (1290KB) ( 393 )   Save BACKGROUND: Human immunodeficiency virus binds to CD4 receptors in human thymocytes, and thus causes activity decreasing in CD4 cells. OBJECTIVE: To explore the relationship between human embryo thymocytes and human immunodeficiency virus. METHODS: Primary cultured human embryo thymocytes were divided into experiment group and control group. Experimental thymocytes were mixed cultured with human immunodeficiency virus 1 Ⅲ B. Control thymocytes were cultured without human immunodeficiency virus. RESULTS AND CONCLUSION: According to transmission electron microscopy, massive human immunodeficiency virus particles emerged in thymocytes treated with human immunodeficiency virus at the 40th hour after cultivation. According to MTT assay and immunohistochemical staining, the absorbance in thymocytes treated with human immunodeficiency virus was significantly lower than that in the control thymocytes at the 40th hour to the 2nd day after cultivation (r=0.9733, P < 0.05). The Fas-L positive rate of the experiment thymocytes treated with human immunodeficiency virus increased on cell membranes and in cytoplasm (P < 0.05). These findings indicate that human immunodeficiency virus decreases the cell activity in primary cultured human embryo thymocytes. Related Articles | Metrics

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Culture and identification of human bladder transitional cells in vitro Hao Wei-ping, Yao You-sheng, Wang Jia-wei, Deng Bi-hua, Lü Yi-song 2011, 15 (50):  9348-9352.  doi: 10.3969/j.issn.1673-8225.2011.50.010 Abstract ( 281 )   PDF (1561KB) ( 493 )   Save BACKGROUND: Methods to separate the transitional epithelial cells include enzyme digestion method, tissue block method and spinning scraping method. OBJECTIVE: To investigate the best approach to culture and passage the human bladder transitional cells in vitro. METHODS: Human bladder transitional epithelial cells were cultured in MEM-F12 and KSFM media. Then the cell morphological changes, growth and proliferation were observed. The bladder transitional epithelial cells were identified by immunofluorescent staining based on the presence of uothelium specific cell markers AE1 and Uroplakin III, the latter one specifically expressed in urothelial tissue. The cells were subcultured by 4-step trypsin digestion method and tissue block replantation method, respectively. RESULTS AND CONCLUSION: The cells in EME-F12 media grew well, and the cell climbing-out and cell fusion occurred earlier than cells in KSFM media. The cells were identified as transitional epithelial cells by immunofluorescence. Passage cells subcultured by trypsin digestion began to adhere to the wall at 18-24 hours after passage. The cells stopped growing after adherence. Cellapoptosis appeared after 60-72 hours. The passage cells subcultured by tissue block replantation displayed a stable and fast growth. The cell began to climb out at 24-48 hours. Cell fusion rate achieved 80% on 10-16 days. Cells began to degenerate at the fourth passage. These results demonstrate that tissue block cultivation and replantation based on DMEM-F12 is an economic and effective way to culture and subculture the human bladder transitional cells. Related Articles | Metrics

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Effect of continuous static hydrostatic pressure on cell proliferation Guo Zi-fen, Feng Yong, Li Kai, Liao Duan-fang 2011, 15 (50):  9353-9355.  doi: 10.3969/j.issn.1673-8225.2011.50.011 Abstract ( 332 )   PDF (742KB) ( 613 )   Save BACKGROUND: Vascular smooth muscle cells grow well in culture condition using static hydrostatic pressure instead of serum. OBJECTIVE: To investigate the effect of continuous static hydrostatic pressure on the proliferation of vascular smooth muscle cells. METHODS: Vascular smooth muscle cell A10 and vascular endothelial cell HUVEC-12 were cultured in a self-developed pressure adjustable incubator, under different pressures of 0, 12, 16, 20, 24 and 28 kPa respectively. RESULTS AND CONCLUSION: The most obvious proliferation of A10 cell was detected under the pressure of 16 kPa, with greatest cell activity. And the most obvious proliferation of HUVEC-12 cell was under 20 kPa pressure, with greatest cell activity. It confirms that limited hydrostatic pressure can promote the proliferation of A10 and HUVEC-12 and increase the cell activity. Related Articles | Metrics

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Combined effects of pulse electric fields and wild-type p53 gene on apoptosis of Hela cells Li Hao-shan, Xiong Zheng-ai, Zhou Wei, Li Cheng-xiang, Yao Chen-guo, Sun Cai-xin 2011, 15 (50):  9356-9359.  doi: 10.3969/j.issn.1673-8225.2011.50.012 Abstract ( 447 )   PDF (1199KB) ( 395 )   Save BACKGROUND: There have been lots of experimental works of animals and cells on pulses electric fields treating tumors. But electric fields combined with gene transfer had been seldom studied. Wild-type p53 gene combined with pulses electric fields (PEFs) can induce apoptosis of carcinoma cells. OBJECTIVE: To investigate apoptosis of Hela cells induced by pulse electric fields combined with wild-type p53 gene and the mutual effect between them. METHODS: PEFs with 100 μs duration, 1 Hz frequency, 8 pieces pulses and 1 500 V/cm electric intensity were performed on Hela cells, Hela-vector and Hela-p53 cells. p53 mRNA expression was detected by RT-PCR. The inhibitory effect on Hela cells growth was detected by MTT assay. Hela cells apoptosis was detected by flow cytometry at 12 hours after PEFs were performed. The expression levels of Bax and Bcl-2 were detected by western blot analysis. Expression of casepase-3 in cells was stained with immunofluorescence and detected by laser scanning confocal microscopy. RESULTS AND CONCLUSION: Apoptotic rate of Hela, Hela-vector and Hela-p53 were (6.48±2.06)%, (7.22±2.25)% and (17.41±2.62)%, respectively. The apoptotic rate was significantly higher in the wild-type p53 gene combined with PEFs group than in the other two groups. At the same time, p53 and caspase-3 mRNA expression was significantly higher in the wild-type p53 gene combined with PEFs group than in the other groups. Bax/Bcl-2 expression ratio was significantly higher in the wild-type p53 gene combined with PEFs group than in the Hela cell group and Hela-vector group as detected by western blot analysis. Hela cells were inhibited by wild-type p53 gene transfer and wild-type p53 gene could increase Hela cells apoptosis induced by PEFs. Related Articles | Metrics

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Construction of a chronic pulmonary thromboembolism animal model Wang Hai-long, Du Zhen-zong, Zhou Zhi-peng, Li Xiang-li, Jiang Yi-yao, Gao Guo-liang 2011, 15 (50):  9360-9364.  doi: 10.3969/j.issn.1673-8225.2011.50.013 Abstract ( 263 )   PDF (1394KB) ( 444 )   Save BACKGROUND: Studies on chronic pulmonary thromboembolism animal models close to the human physiology are rare at home and aboard. OBJECTIVE: To establish a chronic pulmonary thromboembolism animal model by repeated injecting autologous blood clot. METHODS: Rabbits were randomly assigned into embolism group and mock embolism group. Embolism rabbits were repeatedly injected with autologous blood clot through the arteries. Mock embolism rabbits were injected with saline instead. RESULTS AND CONCLUSION: According to the anatomy of pulmonary artery and pathological examination, there was thrombus organization in the 4th week after autologous blood clots injection. CT angiography of pulmonary artery represented signs of local pulmonary artery truncation and some indirect signs, such as inflammation, infarction and pleural thickening. Pulmonary anatomy and pathological examination discovered pulmonary artery thrombus organization and chronic inflammation in the experimental rabbits. These findings demonstrate that chronic pulmonary thromboembolism rabbit model is successfully constructed by repeated injection of autologous blood clot. Related Articles | Metrics

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Stability of a geneogenous scoliosis experimental model accompanied with spinal cord injury Lu Ming, V Ya Fishchenko, Ma Hua-song, Ren Dong-yun, Zheng Rui, Zhang Jing 2011, 15 (50):  9365-9368.  doi: 10.3969/j.issn.1673-8225.2011.50.014 Abstract ( 242 )   PDF (580KB) ( 434 )   Save BACKGROUND: Establishment of a stable and effetive experimental animal model with congenital scoliosis is a precondition to lucubrate the congenital scoliosis with spinal cord injury and an efficient path to analyze the mechanism of spinal cord injury. OBJECTIVE: To assess the stability of congenital scoliosis model accompanied with complex spinal cord injury in rats. METHODS: A number of 15 model rats with no obvious spinal cord symptoms were put into special glass tubes with a diameter of 6 centimeters. The rats were kept in upright position in the tubes for 6 to 10 days to establish the congenital scoliosis model accompanied with complex spinal cord injury. RESULTS AND CONCLUSION: A total of 13 rats showed bilateral hind limb paralysis, adiaphoria of hind limbs and tail and no response to acupuncture and urination disorder. There were no obvious obstacles of defecation. The fur of bilateral hind limbs and tail in all the rats started to wither, became dull or even dropped. The mean Basso, Beattie and Bresnahen score was 7.3±2.2. Modified Tarlov scores were as follows, 0, 5 rats; 1, 2 rats; 2, 4 rats; 3, 1 rat; 4, 1 rat; 5, 0 rat. According to imaging, the congenital scoliosis were all over the spinal cord. The wedged hemivertebra and intercalated disc at the scoliosis vertex moved towards the spinal canal in some rats. According to histochemical results, there was a one-to-one relationship between the congenital defect with bone-and-spinal nourishing vessels damage and the compression in spinal cord segments caused by the bone structure damage in the corresponding segments. There were visible vascular distribution deletion regions at the malformation vertex. The vascular tissue of the venoux plexus underwent apoptosis at the scoliosis vertex. These findings indicate that the symptoms of complex spinal cord injury accompanied with the congenital scoliosis model are similar to the symptoms progress of congenital scoliosis patients. The repeat rate of the rat model is 87% (13/15). Related Articles | Metrics

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Ultrastructural changes of the corticospinal tract in rats after corticospinal tract semitransection Liu Su, Shen Guang-yu, Lü Guang-ming 2011, 15 (50):  9369-9372.  doi: 10.3969/j.issn.1673-8225.2011.50.015 Abstract ( 376 )   PDF (673KB) ( 438 )   Save BACKGROUND: Researches on the overall ultrastructural changes of the body parts near and distant from the corticospinal tract lesion are rarely reported. OBJECTIVE: To investigate the ultrastructural changes of corticospinal tract in rats after corticospinal tract semitransection. METHODS: Rat left vertebral body of medulla was selectively cleavaged to construct corticospinal tract semitransection model. Electron microscopy was utilized to determine the degeneration of the corticospinal tract on the 4th, 14th and 28th days after corticospinal tract semitransection. RESULTS AND CONCLUSION: According to transmission electron microscope observation, the myelin sheath and axons of lesioned corticospinal tract were swelling and irregular in shape. Subsequently, during the extended periods, the ultrastructural changes were myelinoclasis, dissolvation in myelin sheath, and demyelination, shrink of cytoplasm, vacuolar degeneration, increase of organelles in axons progressively. These changes of lesioned corticospinal tract at cervical, thoracic, lumbar segments of spinal cord were more serious than those closed to lesion site. Related Articles | Metrics

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Effects of intrathecal Sufentanil injection on the expression of N-methyl-D-aspartate receptor and calcitonin gene-related peptide in the spinal dorsal horn of rat bone cancer pain models  Wang Yi-chun, Wang Ming-de, Li Zu-rong, Liu Jing-shi, Yang Wen-qian 2011, 15 (50):  9373-9376.  doi: 10.3969/j.issn.1673-8225.2011.50.016 Abstract ( 308 )   PDF (624KB) ( 421 )   Save BACKGROUND: There are considerable animal experiments and clinical evidence indicating obvious analgesic effects of intrathecal Sufentanil injection, but the mechanism behind this is unclear. OBJECTIVE: To investigate the effects of intrathecal Sufentanil injection on the expression of N-methyl-D-aspartate recaptors and calcitonin gene-related peptide in spinal dorsal horn of rat bone cancer pain models. METHODS: A total of 36 healthy male Sprague-Dawley rats were randomly divided into four groups: control group, sham operation group, bone cancer pain model group and Sufentanil group. Bone cancer pain model in the latter two groups was constructed using ascitic fluid of breast cancer cells. Rats in sham operation group were injected with heat inactivated dead cells. Rats in model group and Sufentanil group were treated with saline and Sufentanil daily by intrathecal injection within 15 days after model construction. RESULTS AND CONCLUSION: Compared with control group and sham operation group, the expression of N-methyl-D-aspartate recaptors and calcitonin gene-related peptide in spinal dorsal horn increased in model group on the 12th and 14th days after model construction (P < 0.01). The mechanical allodynia pain threshold and radiant heat stimulus paw withdrawal latency decreased in model group (P < 0.01). Compared with model group, the expression of N-methyl-D-aspartate recaptors and calcitonin gene-related peptide in the superficial layer of spinal dorsal horn decreased on the 12th and 14th days after injection (P < 0.01). The mechanical allodynia pain threshold and radiant heat stimulus paw withdrawal latency increased (P < 0.01). It suggests that intrathecal administration of Sufentanil provide a significant antinociception effect in rat bone cancer pain models, and the mechanisms behind this is related to the inhibition of N-methyl-D-aspartate recaptors and calcitonin gene-related peptide in rat spinal dorsal horn. Related Articles | Metrics

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Preparation of Sprague-Dawley rat models of focal cerebral ischemia-reperfusion injury by suture method Chen Wei-wei, Yang Liu-cai, Pan Shi-wen, Li Shi-hong, Xu Hong-tao 2011, 15 (50):  9377-9380.  doi: 10.3969/j.issn.1673-8225.2011.50.017 Abstract ( 363 )   PDF (719KB) ( 633 )   Save BACKGROUND: There have been no unified standards regarding suture method applied for preparation of focal cerebral ischemia-reperfusion injury models in rats. It is of great significance to search a simple, stable and successful method to prepare focal cerebral ischemia-reperfusion injury models in study of ischemic cerebral disease. OBJECTIVE: To prepare focal cerebral ischemia-reperfusion injury models in Sprague-Dawley rats using suture method and to analyze the success rate and stability of model preparation. METHODS: The conventional Zea Longa model preparation method was modified. Precisely, fishing line was used as the suture line. The proximal parts of external carotid artery and internal carotid artery were ligated. The external carotid artery was not cut off and the pterygopalatine artery was not ligated. The suture line was advanced into the middle cerebral artery via the common carotid artery to cause ischemia and reperfusion. RESULTS AND CONCLUSION: Middle cerebral artery occlusion was performed within 20 minutes. After model preparation, rat neurological functions were obviously impaired. Red tetrazoline staining showed that cerebral infarction region was pale. Pathological examination showed typical pathological manifestation. The success rate of model preparation was 75%. During model preparation, subarachnoid hemorrhage and incomplete cerebral ischemia may occur. These findings suggest that the suture method applied for preparation of focal cerebral ischemia reperfusion injury models in Sprague-Dawley rats is a simple method with high success rate, identified ischemic effects and short operation time.  Related Articles | Metrics

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Corneal histopathological changes and interleukin-6 level in aqueous humor of a rabbit model of penetrating corneal trauma combined with seawater immersion Fan Qin-hua, Chen Qian, Hong Jin, Zhang Zhi-qiang, Yang Yan-qiu 2011, 15 (50):  9381-9384.  doi: 10.3969/j.issn.1673-8225.2011.50.018 Abstract ( 292 )   PDF (497KB) ( 408 )   Save BACKGROUND: Following trauma caused by seawater, cells often exhibit special pathological changes because of the special physico-chemical properties of seawater. OBJECTIVE: To observe corneal histopathological changes and interleukin-6 level in aqueous humor of a rabbit model of penetrating corneal trauma combined with seawater immersion.  METHODS: The rabbit eye models of penetrating corneal trauma caused by firecrackers were established in 16 adult healthy grey rabbits. A 3-mm whole-layer incision was made in the cornea. The right eyes served as experimental sides and the left eyes served as controls. Seawater was injected into the aqueous humor of the right eyes via the corneal incision. The eye surface was flushed with seawater for 30 minutes. Physiological saline was used for the left eyes. RESULTS AND CONCLUSION: Optical microscopy results showed that at 1, 2, 3 days after model establishment, corneal cells on the experimental side exhibited severe necrosis and abscission, obvious swelling of substantia propria layer complicated by cellular infiltration. At 1 and 2 days after model establishment, the pathological changes on the control side were the same as the experimental side, but they were mild, but at 3 and 5 days, they were obviously alleviated. At 1, 2, 3 days after model establishment, interleukin-6 level in aqueous humor was significantly higher on the experimental side than on the control side (P < 0.05). These findings suggest that the degree of injury on the experimental side was more severe than that on the control side, indicating that seawater may be an important causative factor of corneal injury. Related Articles | Metrics

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Establishment of a radial bone critical defect model in a rabbit Zhou Fang, Li Jing, Yu Le2, Liao Hua, Ouyang Jun 2011, 15 (50):  9385-9388.  doi: 10.3969/j.issn.1673-8225.2011.50.019 Abstract ( 365 )   PDF (987KB) ( 437 )   Save BACKGROUND: Nonunion caused by various reasons of fractures and bone defects has been a major problem of clinical orthopedic rehabilitation. With the rapid development of tissue engineering, product quality testing needs standard bone defect model. However, the length of bone critical-size defects has not been ascertained. OBJECTIVE: To establish rabbit radial bone defect models and to determine the length of rabbit radial bone critical-size defects. METHODS: Eighteen male New Zealand white rabbits (2.0-2.5 kg) were randomly assigned into 6 groups and then 12, 13, 14, 15, 16 and 17 mm segmental defects were created in the middle part of radius on both sides, respectively. The incisions were sutured and bandaged, but not fixed. RESULTS AND CONCLUSION: According to examination with naked eyes and X-ray, some of the segmental defects with the length of 12, 13, 14, 15 and 16 mm were completely repaired. But none of the defects with the length of 17 mm was repaired. Histological examination showed that there was trabecular bone formation, bone matrix, medullary cavity recanalization, revascularization and osteoblasts in the segmental defects with the length of 12, 13, 14, 15 and 16 mm. However, there were osteoblasts, osteoclasts in the 17 mm defects, but no medullary cavity recanalization or revascularization. Therefore, the length of bone critical-size defect in rabbit radius was 17 mm. Related Articles | Metrics

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Gene expression of myelin associated protein in adult rats after focal cerebral ischemia and reperfusion Li Hua-jie, Wu Jian, Zhu Lin-feng, Bao Shi-yao 2011, 15 (50):  9389-9392.  doi: 10.3969/j.issn.1673-8225.2011.50.020 Abstract ( 291 )   PDF (1232KB) ( 364 )   Save BACKGROUND: Myelin proteins are the main components in oligodendroglia cells. Research on gene expression changes of myelin related proteins after focal cerebral ischemia and reperfusion will help to understand the role of oligodendroglia cells in focal cerebral ischemia and reperfusion. OBJECTIVE: To explore the gene expression change of myelin related proteins in different cerebral regions and at different time points after focal cerebral ischemia and reperfusion in adult model rats. METHODS: Focal cerebral ischemia and reperfusion model in adult SD rats was constructed using suture method. The early phase mRNA expression changes of myelin proteolipid protein, myelin basic protein and myelin transcription factor 1 in the cerebral cortex regions of infraction center, infarction borders and contralateral area of infarction were detected by fluorescent in situ hybridization with the dig-labeled oligonucleotides. RESULTS AND CONCLUSION: The gene expression of the three myelin related proteins in infraction center decreased significantly in the early phase of focal cerebral ischemia and reperfusion. In infarction borders, there was no significant change in the gene expression of the three myelin related proteins between experiment group and control group on the 1st day after focal cerebral ischemia and reperfusion. Then the gene expression of experiment group increased, and exceeded that of the control group on the 14th day after focal cerebral ischemia and reperfusion (P < 0.05, 0.01); the content of myelin transcription factor 1 positive cells raised the earliest (P < 0.01). The gene expression of cerebral cortex myelin related proteins in infarction borders increased after focal cerebral ischemia and reperfusion in adult SD rats. It indicates that oligodendrocyte precursors are sensitive to cerebral ischemic injury and may involve in the repairing process of cerebral post-ischemic injury. Related Articles | Metrics

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Recombinant human epidermal growth factor combined with alprostadil for treatment of diabetic ulcers Chen Jie-xiang, An Hong-yuan 2011, 15 (50):  9393-9396.  doi: 10.3969/j.issn.1673-8225.2011.50.021 Abstract ( 335 )   PDF (1738KB) ( 444 )   Save BACKGROUND: Previous studies demonstrated that the lack of endogenous recombinant human epidermal growth factor (rhEGF) and the change of hemodynamic parameters will lead to nonunion. OBJECTIVE: To observe the effect of rhEGF combined with alprostadil on diabetic ulcers. METHODS: Forty Wistar rat models of diabetic ulcers were established and randomly divided into four groups: control (1% povidone iodine for debridement), rhEGF (rhEGF gel for spreading), alprostadil (intravenous administration of alprostadil) and rhEGF + alprostadil (rhEGF gel for spreading combined with intravenous administration of alprostadil). RESULTS AND CONCLUSION: At 3, 7, 10 and 14 days after intervention, the area of diabetic ulcers was diminished, healing time was shortened, and dynamic healing rate of wound surface was increased in the alprostadil , rhEGF and alprostadil + rhEGF groups (P < 0.01). The therapeutic effects were superior in the alprostadil + rhEGF group to those in the rhEGF group and alprostadil group (P < 0.01). The therapeutic effects were similar between rhEGF group and alprostadil group. These findings suggest that rhEGF combined with alprostadil better promotes the healing of diabetic ulcers than simple use of rhEGF or alprostadil. Related Articles | Metrics

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Effects of exogenous acidic fibroblast growth factor on the expression of p38MAPK and fibroblast growth-factor receptor 2 in rats with intestinal ischemia-reperfusion injury Weng Li-xin, Li Xiu-xia, Fu Xiao-bing 2011, 15 (50):  9397-9401.  doi: 10.3969/j.issn.1673-8225.2011.50.022 Abstract ( 308 )   PDF (802KB) ( 368 )   Save BACKGROUND: Different types of wound healing can be promoted by acidic fibroblast growth factors. It plays an important role in the repair of visceral injury. OBJECTIVE: To explore the effect of exogenous acidic fibroblast growth factor on the expression of p38MAPK and fibroblast growth-factor receptor 2 in rats with intestinal ischemia-reperfusion injury. METHODS: Rat superior mesenteric artery was microsurgical clipping for 45 minutes to construct intestinal ischemia-reperfusion injury model. Rats were treated with modified acidic fibroblast growth factor immediately after reperfusion. Rat small intestines samples were collected at the 2nd, 6th, 12th and 24th hours after reperfusion to examine. RESULTS AND CONCLUSION: Fibroblast growth-factor receptors 2 were mainly distributed in epithelial cells of small intestinal villi at intestinal cavity, side wall and on the cell membrane of crypts toward cavity. There was no significant change in the expression of fibroblast growth-factor receptor 2 and p38MAPK in protein and mRNA level at the primary stage after reperfusion. Then the expression of fibroblast growth-factor receptor 2 and p38MAPK increased gradually along with the extension of time, and peaked at the 6th to 12th hours after reperfusion. The protein and mRNA expression of fibroblast growth-factor receptor 2 and p38MAPK in rat small intestines was further enhanced after treated with acidic fibroblast growth factor. These findings demonstrate that acidic fibroblast growth factor can promote the repair of intestinal ischemia-reperfusion injury by up-regulating the expression of fibroblast growth-factor receptor 2 and p38MAPK. Related Articles | Metrics

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Temperature changes of surface skin and tympanic membrane in human working muscles during exercise Yi Xue 2011, 15 (50):  9402-9405.  doi: 10.3969/j.issn.1673-8225.2011.50.023 Abstract ( 412 )   PDF (413KB) ( 635 )   Save BACKGROUND: Understanding the body temperature changes during exercise offers new tools to predict human energy metabolism. OBJECTIVE: To explore a new idea on the testing methods of energy metabolism by immediately detecting the human skin temperature and core temperature during the aerobic exercise, in order to suit the requirements of immediately physical detection in modern competitive sports. METHODS: A total of 34 (17males and 17 females) untrained students aged 18-23 years were analyzed. Changes of the skin temperature, core temperature and tympanic membrane were immediately detected under aerobic exercise conditions, respectively. RESULTS AND CONCLUSION: Results showed that the human body core temperature and tympanic membrane temperature did not change significantly during exercise. While the surface skin temperature of working muscles represented a changing trend as follows: temperature dropped in the first two minutes, temperature change was smooth in the 2nd to 4th minutes, temperature raised in the 4th to 6th minutes, the peak temperature appeared at the 6th minute, temperature dropped again in the 6th to 8th minutes, temperature change was smooth in the 8th to 10th minutes and temperature raised again at the 10th minute until to the end. Peak surface skin temperatures of almost all the working muscles appeared at the 6th minute during exercise. It indicates that there is an obvious regular change in the surface temperatures of working muscles during exercise. Related Articles | Metrics

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Static and dynamic plantar pressure analysis by F-Scan three-dimensional dynamic plantar pressure analysis system Fan Xiao-yan, Zhou Jun-jie, Cao Cheng-fu, Chen Xian-qi 2011, 15 (50):  9406-9409.  doi: 10.3969/j.issn.1673-8225.2011.50.024 Abstract ( 433 )   PDF (653KB) ( 959 )   Save BACKGROUND: Former studies indicate that some anatomic regions of feet can not only support the most of the human body weight, but also balance the human body. Massive information concerning the physiology, structure and function of foot, lower limb, and even the whole body can be gained by measuring the peak pressure and distribution of these regions. OBJECTIVE: To observe the dynamic plantar pressure distribution of naturally walking healthy adolescent soccer players, such as peak power values of both feet, loading impulse. METHODS: The dynamic plantar pressure of 16 to 19 years old adolescent soccer players were tested using the F-Scan Gait three-dimensional dynamic plantar pressure analysis system. RESULTS AND CONCLUSION：Plantar stress-time curves showed a clear double-peak type. The mean peak force value of the 10 tested plantar region all reached maximum in the lateral heel (P < 0.01). For the ones preferred right foot, the mean peak force value of great toe, the first and second metatarsals, the inner and outer heel regions of the right foot was larger than that of the left foot (P < 0.05). While the mean peak value of the fifth metatarsal, two foot arch regions of the left foot was larger than that of the right foot (P < 0.05). During walking, the load centers in the whole foot contact phase and the outside-ground phase were the first and second metatarsals, the inner and outer heel regions. The maximum load parts of foot were the first and second metatarsals, the inner and outer heel regions. The distribution regularity of left foot was similar to that of the right foot, and there was no difference between genders. In conclusion, there is a consistency in the distribution regularities of the left and right foot static and dynamic plantar pressure analysis in the healthy adolescent soccer players. Related Articles | Metrics

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Electro-acupuncture treatment inhibits inflammatory reaction and postpones articular cartilage degeneration in osteoarthritis Wu Ming-xia, Li Xi-hai, Wu Guang-wen, Li Li, Chen Wen-lie, Liu Xian-xiang 2011, 15 (50):  9410-9414.  doi: 10.3969/j.issn.1673-8225.2011.50.025 Abstract ( 355 )   PDF (932KB) ( 467 )   Save BACKGROUND: Several studies have shown that electro-acupuncture (EA) can effectively alleviate the clinical symptoms of osteoarthritis. OBJECTIVE: To investigate the mechanism by which EA treatment inhibits osteoarthritis inflammation and protects the function of articular cartilage. METHODS: SD rat models of osteoarthritis were established by double knee joint cavity injection of papain. Osteoarthritis rat models were treated with EA at EX-LE4 and EX-LE5 for 5, 15 and 30 minutes per day. RESULTS AND CONCLUSION: Compared with the model group, substance P in blood and interleukin 1, interleukin 6, tumor necrosis factor α, matrix metalloproteinase 3 concentrations in synovial fluid were significantly reduced, leptin in blood and matrix metalloproteinase 1 in synovial fluid were significantly increased (P < 0.05), and the degree of cartilage degeneration was significantly alleviated (P < 0.05, P < 0.01) in osteoarthritis rat models after treatment with EA for 15 and 30 minutes per day  (P < 0.05). These results showed that EA can effectively alleviate the inflammatory reaction and postpone articular cartilage degeneration in osteoarthritis.  References | Related Articles | Metrics

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Effect of long needle penetration needling on matrix metalloproteinase 3 in the synovium of knee osteoarthritis model rats Li Shi-sheng, Wu Yao-chi, Zhang Jun-feng, Zhou Jing-hui, Sun Yi-jun, Huang Cheng-fei 2011, 15 (50):  9415-9418.  doi: 10.3969/j.issn.1673-8225.2011.50.026 Abstract ( 344 )   PDF (633KB) ( 625 )   Save BACKGROUND: Studies have shown that matrix metalloproteinases 3 plays an important role in articular cartilage degeneration and destruction. OBJECTIVE: To observe the changes of metalloproteinases 3 expression level in synovium of knee osteoarthritis rats after treated with penetration needling of long needle and sodium hyaluronate. METHODS: SD rats were injected with 1.6% papain solution to knee joint cavity and were driven to exercise in order to construct knee osteoarthritis model. Rats were randomly assigned into 3 groups in the 2nd week after model construction: penetration needling group, drug group and model group. Penetration needling group were treated by penetration needling with filiform needle, with a diameter of 0.30 millimeters and a length of 125 milimeters. Drug group were injected with sodium hyaluronate to knee joint cavity. Model group received no treatment. And non-model control group were established. RESULTS AND CONCLUSION: Content of synovium matrix metalloproteinase 3 in model group was significantly increased than that in the control group (P < 0.05) in the 4th week after treatment. The content of matrix metalloproteinase 3 in penetration needling group and drug group was significantly reduced than that in the model group (P < 0.05). Penetration needling group was similar to drug group (P > 0.05). These findings demonstrate that long needle penetration therapy can effectively correct the abnormal expression of matrix metalloproteinase 3 in synovium of knee osteoarthritis model rats. It may be one of the mechanisms of the treatment of knee osteoarthritis by long needle penetration needling. Related Articles | Metrics

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Construction and identification of recombinant adeno-associated virus vector with human bone morphogenetic protein 4/7 fusion gene Yuan Shao-hui, Liu Wei, Wu Bin-qi, Han Xi-guang, Bi Zheng-gang 2011, 15 (50):  9419-94.  doi: 10.3969/j.issn.1673-8225.2011.50.027 Abstract ( 473 )   PDF (769KB) ( 343 )   Save BACKGROUND: Studies have shown that the activity of bone morphogenetic protein (BMP) heterodimer is 20 times higher than that of BMP homodimer, but related BMP 4/7 fusion gene-associated virus vector has been rarely reported. OBJECTIVE: To construct the recombinant adeno-associated virus vector with human BMP 4/7 fusion gene (AAV-BMP-4/7) and to detect its expression. METHODS: Human BMP 4, 7 mature peptide gene was amplified. Plasmids pGEM-human BMP 4, 7 were constructed, respectively. BMP 4/7 fusion gene was harvested using DNA ligation and then cloned to harvest pGEM-BMP-4/7 plasmid for gene sequencing. HEK293 cells were transfected with adeno-associated virus (AAV) particles to harvest viral vector AAV-BMP-4/7. BMP-4/7 fusion gene protein expression in E. coli was detected by SDS-PAGE. AAV-BMP-4/7 of different multiplicities of infection (MOI) was used to transfect bone marrow stromal cells (BMSCs) for 3 days. Total cellular RNA and total cellular protein were extracted. BMP-4/7 fusion gene expression in BMSCs was detected by RT-PCR and ELISA. RESULTS AND CONCLUSION: The constructed recombinant pSNAV-BMP-4/7 was confirmed by PCR electrophoresis and digestion with restriction enzyme. The gene sequence in the recombinant pSNAV-BMP-4/7 was correct. Ttranscription bands of BMP-4/7 fusion gene in BMSCs were detected by RT-PCR. ELISA showed that BMP-4/7 protein in the AAV-transfected BMSCs was gradually increased with the increase in MOI (P < 0.01). These results showed that the AAV-BMP-4/7 was successfully constructed. Related Articles | Metrics

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Construction of a eukaryotic expression vector containing mouse heat shock transcription factor 1 Ouyang Hua-wei1, Tan Jun1, Li Gao-feng1, Luo Cheng-qun2 2011, 15 (50):  9425-9427.  doi: 10.3969/j.issn.1673-8225.2011.50.028 Abstract ( 248 )   PDF (528KB) ( 409 )   Save BACKGROUND: Heat shock transcription factor 1 has heat shock response effect. OBIECTIVE: To construct a eukaryotic expression vector containing mouse heat shock transcription factor 1 (HSF1). METHODS: Amplified mouse HSF1 cDNA fragment was cloned into pcDNA3.1 to construct a eukaryotic expression vector pcDNA3.1-HSF1. RESULTS AND CONCLUSION: There were two fragments in electrophoresis after restriction enzyme digestion, 5.5 kb fragment and 1.5 kb fragment. The result of DNA sequencing showed that the sequence of the cloned HSF1 was identical to that GeneBank had reported. HSF1 protein band with the relative molecular mass of 72 000 was detected by western blot. These findings indicate that the eukaryotic expression vector pcDNA3.1-HSF1 containing mouse HSF1 is successfully constructed. Related Articles | Metrics

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Construction and expression of Ad-EGFP-MCP-1 vector Zhang Fang, Wang Wei, Xie Yue, Zhang Hao 2011, 15 (50):  9428-943.  doi: 10.3969/j.issn.1673-8225.2011.50.029 Abstract ( 246 )   PDF (794KB) ( 369 )   Save BACKGROUND: Monocyte chemotactic factor can promote arterialization of newly formed vessels to form functional arteriole and then realize angiogenesis. OBJECTIVE: To construct Ad-EGFP-MCP-1 vector and to observe its expression in vivo after package, purification and viral titer detection. METHODS: MCP-1 gene was amplified by PCR and the sequence was compared with Genebank data, and then sub-cloned into the pDC315-EGFP vector after sequence analysis. Ad-EGFP-MCP-1 was obtained with pBHG lox ΔE1,3 Cre secondary packaging system. After transfection of Ad-EGFP-MCP-1 via trachea, EGFP expression was observed under fluorescence microscope.  RESULTS AND CONCLUSION: The full-length of MCP-1 was obtained by PCR and identified by sequencing. Ad-EGFP-MCP-1 recombinant adenovirus vector was stably expressed in rat lung tissue. EGFP expression reached the peak level at 1 week and maintained this level for approximately 4 weeks. Ad-EGFP-MCP-1 recombinant adenovirus vector was successfully constructed, packaged and amplified, and it is effectively expressed in the lung tissue. Related Articles | Metrics

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Construction and identification of endothelial cell lines stably expressing human heat shock protein 22 Bao Hui-hui, Chen Qi, Wang Ling, Cheng Xiao-shu 2011, 15 (50):  9433-9436.  doi: 10.3969/j.issn.1673-8225.2011.50.030 Abstract ( 294 )   PDF (535KB) ( 275 )   Save BACKGROUND: Stress expression of heat shock protein 22 (HSP22) may protect endothelial cells against hypoxia/reoxygenation injury. OBJECTIVE: To establish endothelial cell line stably expressing human HSP22 gene. METHODS: The eukaryotic expression vector pEGFP-N1-HSP22 was identified by EcoRⅠ/kpnⅠdouble enzyme digestion. The recombinant plasmids were transfected into human umbilical vein endothelial cells (HUVECs) by lipofectamine. After screening culture by G418, a stably transfected cell line was established. The transcription and expression of the HSP22 gene were identified by fluorescence microscope, RT-PCR and western-blot assay. RESULTS AND CONCLUSION: Agarose gel electrophoresis detection showed that HSP22 DNA segment was in accordance with the expected results after enzyme digestion. Green fluorescent protein expression was observed in each cell through the use of fluorescence microscope. HSP22 mRNA and protein expression were detected by RT-PCR and western blot analysis. The HUVEC monoclonal cells with a stable expression of HSP22 gene have been established successfully. Related Articles | Metrics

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Plasmid construction and variation analysis of exon-4 of carnitine palmitoyltransferase-Ⅱ gene Yao Deng-bing, Yao Min, Wang Li, Dong Zhi-zhen, Yao Deng-fu 2011, 15 (50):  9437-9440.  doi: 10.3969/j.issn.1673-8225.2011.50.031 Abstract ( 468 )   PDF (673KB) ( 589 )   Save BACKGROUND: Carnitine palmitoyltransferase-Ⅱ (CPT-Ⅱ) is located in the inner mitochondrial membrane and is one of the pivotal enzymes during the fatty acid oxidation. Currently, there are no studies about influenza virus infection and variation of CPT-Ⅱ gene. OBJECTIVE: To construct the plasmid expressing exon-4 of CPT-Ⅱ gene (CPT-Ⅱ-E4) and analyze its mutation. METHODS: Human CPT-Ⅱ DNA from the blood of two patients infected with influenza virus was involved. After the PCR amplification, the product of CPT-Ⅱ-E4 was cloned into pGEMD-T vector. The T-CPT-ⅡE4 was transferred to E. coli DH5α cells after prepared with T4 DNA higase. Then propagation of bacteria, preparation of plasmid, EcoRI digestion, sequencing, and variation analysis were performed. RESULTS AND CONCLUSION: The recombinant pGEM-T-CPT-ⅡE4 was proved to contain whole sequence of CPT-Ⅱ-E4 with 1305 nucleotides which could code 435 amino acids. According to the original sequence from Genebank, two variable sites were found at 1618 (GTC→ATC) and 1858 (TTT→TCT) which code amino acids of V368I and F448L respectively. Related Articles | Metrics

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ROCK1 and ROCK2 silencing by ShRNA in rat cardiomyocytes Ding Hao, Li Ju-xiang, Sun Guo-fang, Hong Kui, Cheng Xiao-shu 2011, 15 (50):  9441-9444.  doi: 10.3969/j.issn.1673-8225.2011.50.032 Abstract ( 293 )   PDF (625KB) ( 372 )   Save BACKGROUND: Previous studies have proved that Rho-associated coiled-coil protein kinase (ROCK) 1 and ROCK2 involve in the process of apoptosis. On that basis, a hypothesis has been proposed that myocardial apoptosis can be inhibited by down-regulating the expression of ROCK1 and ROCK2. OBJECTIVE: To verify whether the transfection of ROCK1-short hairpin RNA (ShRNA) and ROCK2-shRNA can effectively silence the expressions of ROCK1 and ROCK2 in rat cardiomyocytes and to screen for the shRNA with the best silencing effeciency. METHODS: Three specific recombinant plasmids of ROCK1-shRNA and ROCK2-shRNA were constructed and identified respectively. Silencing effects of ROCK1 and ROCK2 in SD rat myocardial cells were assessed after shRNA transfection. And the shRNA with the best silencing efficiency was selected among these shRNAs. RESULTS AND CONCLUSIONS: Western blot results showed that, compared with the control groups, the transfection of all three recombinant plasmids of ROCK1-shRNAs and ROCK2-shRNAs inhibited the expressions of ROCK1 and ROCK2 significantly (P < 0.05 or P < 0.01). Among these shRNAs, ROCK1-shRNA1 and ROCK2-shRNA2 showed the best silencing efficiency of (82.15±7.53)% and (89.42±5.81)%, respectively (P < 0.05). These findings show that the expressions of ROCK1 and ROCK2 in rat cardiomyocytes can be effectively inhibited by the transfection of ROCK1-shRNA and ROCK2-shRNA. And the shRNA with the best silencing efficiency was selected among these shRNAs. Related Articles | Metrics

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Expression of peroxidase 6 in fetal rat testicular cells in pregnant rat hypospadias model induced by di-n-butyl phthalate Shen Bai-xin, Wei Zhong-qing, Ju Xiao-bing, Zuo Wei, He Jia, Shi Hua-chao, Zhang Wei, Wang Xin-ru 2011, 15 (50):  9445-9448.  doi: 10.3969/j.issn.1673-8225.2011.50.033 Abstract ( 327 )   PDF (1391KB) ( 510 )   Save BACKGROUND: Previous study shows that expression of peroxidase 6 in fetal rat testis increase significantly induced by in utero exposure to di-n-butyl phthalate. OBJECTIVE: To further investigate the effect of prenatal di-n-butyl phthalate exposure on peroxidase 6 expression. METHODS: Rats of 14 to 18 days of pregnancy were exposed to di-n-butyl phthalate with the dose of 800mg / (kg•d). Testes were harvested from the fetal rats from gestational day 21 and peroxidase 6 was extracted. The expression of peroxidase 6 was detected and analyzed by western blot and immunohistochemical methods. RESULTS AND CONCLUSION: According to western blot, the expression of peroxidase 6 in experiment group (0.205±0.020) was significantly higher than that in the control group (0.110 ± 0.020, P < 0.05). According to immunohistochemistry, peroxidase 6 was mainly localized in Leydig cells of fetal rat testes. These findings indicate that peroxidase 6 may antagonize di-n-butyl phthalate toxicity by antioxidation and stimulating the proliferation of Leydig cells. The function of peroxidase 6 needs further research. Related Articles | Metrics

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Antarctic krill trypsin promotes wound healing Zhang Peng, Jiang Guo-liang, Wang Jun-ren 2011, 15 (50):  9449-9452.  doi: 10.3969/j.issn.1673-8225.2011.50.034 Abstract ( 466 )   PDF (1370KB) ( 355 )   Save BACKGROUND: It is confirmed that trypsin has better effects on debridement and wound healing. OBJECTIVE: To explore the effects of the Antarctic krill trypsin on wound healing. METHODS: Back trauma model in New Zealand white rabbits was established by defecting full-thickness skin. There were four wounds in each rabbit, treated with crude trypsin, pure trypsin extracted from Antarctic krill, papin and saline, respectively. The wounds were treated twice daily until healed. RESULTS AND CONCLUSION: According to the healing area at the 5th, 10th and 15th days after treatment, crude trypsin and pure trypsin extracted from Antarctic krill had a better effect on wound healing. Crude trypsin was better than pure trypsin. The residual wounds area of crude typsin group for each time period was smaller than that of saline group and papain group (P < 0.05). The content determination results of skin hydroxyproline showed that crude trypsin significantly improved the synthesis and accumulation of collagen in wound tissue. According to histological sections, crude typsin group achieved the best efficiency of the treatment. These results demonstrate that Antarctic krill trypsin can promote wound healing in New Zealand white rabbits, and the crude typsin has the best effect. Related Articles | Metrics

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Expression of Toll-like receptor 4 in rat periodontal ligament during orthodontic tooth movement Nie Jing, Zhong Liang-jun, Li Xiao-Bing, Lin Bao-Shan, Qian Xu, Qian Ya-Jing 2011, 15 (50):  9453-9456.  doi: 10.3969/j.issn.1673-8225.2011.50.035 Abstract ( 350 )   PDF (973KB) ( 396 )   Save BACKGROUND: Extensive research has been done on the function of the periodontal ligament under the orthodontic force. However, there are no reports regarding the expression of Toll-like receptor 4 in rat periodontium during orthodontic tooth movement. OBJECTIVE: To investigate the expression of Toll-like receptor 4 in rat periodontium during orthodontic tooth movement. METHODS: Rat orthodontic tooth movement model was constructed using NiTi extension spring with the force value of 50 g. The orthodontic periodontium was collected on the 1st, 3rd, 5th, 7th, 10th and 14th days after exerting force to detect the expression of Toll-like receptor 4. RESULTS AND CONCLUSION: According to immunofluorescence staining, the expression of Toll-like receptor 4 in the periodontal tissues increased on the 1st day after exerting force, and reached highest level on the 5th day; then fell to the initial level on the 14th day. Thus the expression of Toll-like receptor 4 is evoked by orthodontic force, and it may play a role in periodontal remodeling. Related Articles | Metrics

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Early application of sodium tanshinoneⅡA sulfonate can depress hypertrophic scar of rabbit ear Zhang Jian-hua, Ou Bin-xian, Meng Cheng-yue 2011, 15 (50):  9457-9461.  doi: 10.3969/j.issn.1673-8225.2011.50.036 Abstract ( 258 )   PDF (1497KB) ( 496 )   Save BACKGROUND: Sodium tanshinoneⅡA sulfonate (STS) play effects on anti-oxidant, and depression of fibration.  OBJECTIVE: To investigate the effect of early injection of STS on hypertrophic scar (HS) of rabbit ear. METHODS: Scar model were established by removing the skin and perichondrium in rabbits ears on facies ventralis. STS    0.05 mg, 0.1 mg, 0.2 mg (or 40μl normal saline, respectively, experimental groups), normal saline 40 μL(control group) were injected into early rabbits ears scars, once a week, 3 weeks all together. RESULTS AND CONCLUSION: After 3 weeks STS injection, the hyperplasia and the thickness were decreased; the quantity of collagen fiber and vascular in scar tissue section was decreased in experimental group detected by hematoxylin-eosin staining. The collagen fiber area in scar tissue section was decreased by Masson staining and the amount and the volume of fibroblast were decreased under electron microscope. That indicated that early STS injection could depress HS in rabbits scar ears. Related Articles | Metrics

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Effect of astragalus polysaccharide on cellular immunity of rats with recurrent aphthous ulcer Wang Xue-mei, Bo Lei, Li Xin, Zhang Ju-mei, Yu Zhan-hai 2011, 15 (50):  9462-9465.  doi: 10.3969/j.issn.1673-8225.2011.50.037 Abstract ( 376 )   PDF (687KB) ( 423 )   Save BACKGROUND: Recurrent aphthous ulcer is caused by complicated factors. It may be related to the imbalance of immune regulation. OBJECTIVE: To investigate the relationship between the abnormal cellular immune and the pathogenesis of recurrent aphthous ulcer and to explore the immune regulation of astragalus polysaccharide in rats with recurrent aphthous ulcer. METHODS: Immunization method was used to establish recurrent aphthous ulcer model in SD rats. Rats were treated with 100, 200 and 300 mg/kg astragalus polysaccharide or saline by gavage after model establishment. Rats receiving normal feeding were used as control. The values of peripheral blood T lymphocyte subsets CD4+, CD8+ and CD4+/CD8+ in SD rats of each group were measured by flow cytometry. RESULTS AND CONCLUSION: According to flow cytometry, the values of peripheral blood CD4+ and CD4+/CD8+ decreased significantly in rats with recurrent aphthous ulcer, while the value of CD8+ increased significantly (P < 0.05). On the 20th day after treated with 100, 200 and 300 mg/kg astragalus polysaccharide by gavage, the values of peripheral blood CD4+ and CD4+/CD8+ partially recovered in rats with recurrent aphthous ulcer, expecially in the rats treated with 300 mg/kg astragalus polysaccharide. It demonstrates that the unbalance of T-lymphocyte is related to the pathogenesis of recurrent aphthous ulcer. Astragalus polysaccharide can dose-dependently improve the abnormal immune state in rats with recurrent aphthous ulcer. Related Articles | Metrics

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Research progress in autophagy for mass maintenance of skeletal muscle Wang Gui-ping 2011, 15 (50):  9468-9472.  doi: 10.3969/j.issn.1673-8225.2011.50.039 Abstract ( 344 )   PDF (858KB) ( 529 )   Save BACKGROUND: Mass maintenance of skeletal muscle plays essential roles in prevention and treatment of muscle disorders, debilitation and weakness. The regulation and role of autophagic pathway in protein of skeletal muscle are the research frontiers all over the world, which offer new ideas and thoughts for the further study of muscle mass maintenance. OBJECTIVE: To discuss both the advantages and disadvantages of autophagic signaling transduction pathway in metabolic remodeling of catabolism in skeletal muscle. METHODS: An online search was performed by the ISI Web of knowledge database (1970/2010) with key words “autophagy, skeletal muscle, protein, muscle atrophy” in English. The functions of autophagic signaling pathway in muscle, loss mass maintenance and metabolic remodeling were summarized, and the research progress in autophagy-lysosome systems was analyzed. RESULTS AND CONCLUSION: According to inclusion and exclusion criteria, 64 papers were chosen. The activation of autophagic signaling system contributes to the damage of the myoprotein and the progress of muscle atrophy, but excessive autophagy like ubiquitin-proteasome system do have negative impacts on muscle mass. At the same time, autophagic signaling system also contributes to maintaining steady-state in muscle fibers. In addition, autophagic system in skeletal muscle is different from other metabolisms, and has two kinds of progresses, including short and long items, which are controlled by two different signaling systems. Therefore, the remodeling mechanism of autophagic signaling system needs to be further studied in the future. Related Articles | Metrics

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Effects of creatine supplementation on skeletal muscle energy metabolism in old adults Jiang Yu-xing, Wang Xu-lou, Gao Qian-jin 2011, 15 (50):  9473-9476.  doi: 10.3969/j.issn.1673-8225.2011.50.040 Abstract ( 517 )   PDF (604KB) ( 527 )   Save BACKGROUND: Research shows that creatine supplementation is a safe and effective method to diminish age-related declines in muscle mass and strength. OBJECTIVE: To summarize studies about the effects of creatine supplementation on skeletal muscle energy metabolism, exercise capacity and life quality in old adults. METHODS: An online search of PubMed database and CNKI was performed using key words of “creatine; old adults; skeletal muscle; phosphocreatine; strength” for articles published before March 2011, both in English and Chinese. Papers about effects of creatine supplementation on skeletal muscle energy metabolism and exercise capacity in old adults were included, and repetitive researches were excluded. RESULTS AND CONCLUSION: A total of 72 documents were retrieved, and 38 articles were retained after depleting unrelated and repetitive ones. The recent researches concerning the effects of creatine supplementation on intramuscular energy metabolism, skeletal muscle morphology and life quality in old adults were summarized. Creatine can increase muscle strength, induce muscle hypertrophy and improve the exercise capacity of old adults. Creatine supplementation is a safe, inexpensive and effective nutritional intervention. It can slow the muscle wasting rate associated with aging, especially when consumed in conjunction with resistance training. Related Articles | Metrics

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Research status and effect of muscle tissue engineering on exercise-induced muscle injury Wang Peng-ju 2011, 15 (50):  9477-9480.  doi: 10.3969/j.issn.1673-8225.2011.50.041 Abstract ( 434 )   PDF (670KB) ( 454 )   Save BACKGROUND: High levels of fast-paced sports game is likely to cause muscle damage which is hopeful to repair because of muscle tissue engineering. OBJECTICE: To review the research status of muscle tissue engineering for tissue engineering exercise-induced muscle injury studies provide a theoretical basis. METHODS: The articles from PubMed database between January 1994 and December 2010 were retrieved by computer with the key words of “tissue engineering, anterior cruciate ligament repair, biological material scaffold, cytokines, seed cells.” Articles related to exercise-induced muscle damage and muscle tissue engineering were included. Meta-analysis an duplicate studies were excluded. RESULTS AND CONCLUSION: A total of 15 literatures were included. Current researches projects in four areas include rebuilding muscle tissue, extracellular matrix composite materials, muscle tissue engineering seed cells sources, matrix scaffold. Current researches show that exercise-induced muscle tissue engineering for the treatment of muscle injuries has a good prospect. However, in practice applications, muscle tissue-engineered seed cells selection, scaffold construction, extracellular matrix composite materials research and organizational compatibility are the still the difficult problems of muscle tissue engineering. Related Articles | Metrics

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Relationship between training cessation and neural regulation of muscle strength Song Ya-jun 2011, 15 (50):  9481-9485.  doi: 10.3969/j.issn.1673-8225.2011.50.042 Abstract ( 419 )   PDF (581KB) ( 651 )   Save BACKGROUND: Training cessation usually occurs when athletes encounter certain circumstances, such as disease, injury or travel. This might affect the neural regulation of skeletal muscle. OBJECTIVE: To fully understand the effect of training cessation on muscle strength neural regulation ability and the mechanism behind it. METHODS: An online search of Medline database was performed for relevant reviews and research papers on training cessation and neural regulation of muscle strength published between 1983 and 2006. Then a tracing search was performed according to the references of the collected articles. RESULTS AND CONCLUSION: A total of 48 articles on training cessation and neural regulation of muscle strength were collected. The training cessation with certain duration can cause the decrease of maximal voluntary contraction, but this might be affected by spinal cord, nerve centre above the spinal cord and skeletal muscle itself. In the mean time, the maximal voluntary contraction and corresponding neural adaptation may retain to some extent during the training cessation, depending on the exercise training mode, the muscle contraction pattern, the testing position and circumstance during training cessation. Related Articles | Metrics

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Correlation between Vitamin D receptor gene polymorphism and osteoporosis Luan Jun-wei, Fan Xue-hui, Chen Zhan-wen 2011, 15 (50):  9486-9490.  doi: 10.3969/j.issn.1673-8225.2011.50.043 Abstract ( 283 )   PDF (835KB) ( 470 )   Save BACKGROUND: Osteoporosis is a polygenic disease, and the peak bone mass and bone mass loss are significantly determined by genetics factors. OBJECTIVE: To determine the genotype frequencies of Vitamin D receptor gene ApaⅠ polymorphism and its association with osteoporosis of han population in Shandong Peninsula, and then to approach the predisposing factors of primary osteoporosis. METHODS: A total of 367 han people coming from unrelated familes who lived in Shandong Peninsula for many years were selected. The subjects were divided into normal bone mineral density group (n=227), osteoporosis group (n=63) and ospteoporosis with osteoporotic fracture group (n=77). RESULTS AND CONCLUSION: The genotype frequency distribution of Vitamin D receptor gene followed the Hardy-Weinberg equilibrium (χ2 =1.583, P > 0.05). The frequency distribution of genotypes aa, Aa and AA were 53.1%, 10.6 % and 36.3%, respectively. Body mass index had significant positive correlation with bone mineral density (P < 0.01), while age had significant negative correlation with bone mineral density (P < 0.01). People with aa genotype had significant lower bone mineral density in lumbar spine and ward’s triangle (P < 0. 05) after correcting the body mass index and age. According to χ2 test, there was no significant difference between osteoporotic fracture group and normal control group (χ2 =4.795,P > 0.05). The experimental results show that there is possible correlation between polymorphisms of restriction sites in Vitamin D receptor gene Apa Ⅰ and bone mineral density in han people of Shandong Peninsula. It indicates that the restriction site polymorphisms of Apa Ⅰ gene may be used as genetic markers in predicting the risk of developing osteoporosis. Related Articles | Metrics

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Polymorphism of G protein β3 subunit 825C/T in Korean population and Han population with essential hypertension in Mudanjiang region Song Jie, Lu Shuang, Gao Xiao-hong, Zhang Shu-jie 2011, 15 (50):  9491-9495.  doi: 10.3969/j.issn.1673-8225.2011.50.044 Abstract ( 372 )   PDF (747KB) ( 438 )   Save BACKGROUND: Whether there is a relationship between the gene polymorphisms of G-protein β3 subunit 825 C/T and the primary hypertension of Korean population remains unanswered. OBJECTIVE: To explore the gene polymorphisms of C825T in Korean population and Han population with essential hypertensive in Mudanjiang. METHODS: A total of 224 essential hypertensive patients receiving medical serving in the Internal Medicine Department of Koreans Hospital of Mudanjiang City were selected as hypertension group. A number of 196 healthy people underwent physical examinations in the corresponding period at the clinic were selected as normotensive group. PCR-RFLP method was used to measure the polymorphism of G-protein β3 subunit C825T. Genotype frequencies and allele frequencies of TT, TC and CC were investigated. RESULTS AND CONCLUSION: PCR-RFLP results demonstrated that with respect to Korean population in Mudanjiang, there was no significant difference in the genotype frequencies of TT, TC and CC between the hypertension group and normotensive group (P < 0. 05), and there was significant difference in the allele frequencies of T and C between these two groups (P < 0. 05). With respect to Han population in Mudanjiang, there was significant difference in the genotype frequencies of TT, TC and CC between the hypertension group and normotensive group (P < 0.05), but no significant differences in allele frequencies of T and C between these two groups (P > 0.05). Allele frequency of C in hypertension group of Korean population was higher while the allele frequency of T was higher in hypertension group of Han population. There was a difference between nations. These findings indicate that the gene polymorphisms of G-protein β3 subunit C825T is not related to hypertension of Korean population in Mudanjiang, but is a risk factor of hypertension in Mudanjiang Han population. Related Articles | Metrics

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Relations between the expression level of FOXC2 mRNA in ventral adipose tissue and type 2 diabetes Nian Xin, Wei Jun-jie, Su Yan-dan, Liu Hua, Zhang Xu-xiang, Li Hong 2011, 15 (50):  9496-9500.  doi: 10.3969/j.issn.1673-8225.2011.50.045 Abstract ( 408 )   PDF (613KB) ( 790 )   Save BACKGROUND: FOXC2 is closely related to the differentiation of adipose cells. It plays an important role in many aspects, such as energy metabolism, resistance of type II diabetes and metabolic syndrome. OBJECTIVE: To investigate the expression level of FOXC2 mRNA in human ventral adipose tissue of Yunnan Province, and to explore the relations between FOXC2 and type II diabetes. METHODS: Type II diabetic patients or nondiabetic patients underwent abdominal surgery in Affiliated Hospital of Kunming Medical University were randomly selected as experimental subjects, with 50 cases in each group. The expression level of FOXC2 mRNA in ventral adipose tissue was detected by RT-PCR. The correlation of FOXC2 mRNA expression and subject clinical data was analyzed. RESULTS AND CONCLUSION: The expression level of FOXC2 mRNA in ventral adipose tissue in type II diabetic patients decreased. The FOXC2 mRNA expression level of diabetic group was inversely related to fasting blood-glucose and blood pressure. The FOXC2 mRNA expression level of nondiabetic group was inversely related to fasting blood-glucose. These findings indicate that FOXC2 may reduce insulin resistance and improve the regulation of insulin on sugar metabolism in type II diabetic patients. In addition, the decrease of FOXC2 expression shows a synergetic effect on blood press raise in diabetic patients. Related Articles | Metrics