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11 June 2013, Volume 17 Issue 24 Previous Issue    Next Issue

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Culture of bone marrow mesenchymal stem cells plus allogenic decalcified bone matrix in the knee cavity of rabbits for tissue engineered cartilage 2013, 17 (24):  4371-4379.  doi: 10.3969/j.issn.2095-4344.2013.24.001 Abstract ( 423 )   PDF (2658KB) ( 515 )   Save BACKGROUND: It remains controversial how to repair articular cartilage defects and achieve a good long-term efficacy using tissue engineering methods. OBJECTIVE: To investigate the feasibility of culturing rabbit bone marrow mesenchymal stem cells in combination with allogenic decalcified bone matrix in the knee cavity of rabbits for constructing tissue engineered cartilage.  METHODS: Bone marrow mesenchymal stem cells were isolated using adherence screening method and cultured in complete Dulbecco’s modified Eagle’s medium/Ham’s nutrient mixture F-12. The third generation was induced to chondrocytes. Allogenic decalcified bone matrix, made from rabbit’s ilium and vertebral bone, was combined with the induced bone marrow mesenchymal stem cells in vitro for 1 day. The cell-scaffold composites wrapped by fascia were cultured in the left knee cavities of adult rabbits (cell-scaffold group, n=15) while simple homologous decalcified bone matrix wrapped by fascia were cultured in the right knee cavities (scaffold group, n=15). The specimens were harvested at 4, 8, and 12 weeks after culture for gross and histological observations. Histological observation was conducted using hematoxylin-eosin staining, toluidine blue staining and immunohistochemical staining.  RESULTS AND CONCLUSION: At 4 and 8 weeks after culture, the mean absorbance value was respectively 0.263±0.031 and 0.340±0.052 for the cell-scaffold group and 0.147±0.027 and 0.165±0.030 for the scaffold group, with significant differences between two groups (P < 0.05). At 12 weeks, the mean absorbance value of specimens in the cell-scaffold group was 0.362±0.037, and the samples exhibited similar appearance of normal cartilage and positive immunohistochemical reaction of type II collagen. However, the scaffold degraded in the scaffold group. At 12 weeks, hematoxylin-eosin staining revealed increased chondrocytes and mostly absorbed decalcified bone matrix scaffold in the cell-scaffold group while toluidine blue staining showed metachromasia matrix dyed red or purple. Results indicate that tissue engineered cartilage can be induced by culturing rabbit bone marrow-derived mesenchymal stem cells combined with allograft decalcified bone matrix in the knee cavity of rabbit. Figures and Tables | References | Related Articles | Metrics

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Receptor activator of nuclear factor kappa B ligand-induced mature osteoclasts Chen Guo-xian, Wang Guo-rong, Lin Zong-jin, Li Guo-shan, Guo Chun-xian, Luo Yuan-biao, 2013, 17 (24):  4380-4385.  doi: 10.3969/j.issn.2095-4344.2013.24.002 Abstract ( 474 )   PDF (1435KB) ( 499 )   Save BACKGROUND: Osteoclast as a terminal cell is difficult to obtain, and the shortage of mature osteoclast strains limits its application. Previous studies have shown that stromal cell culture or co-culture is commonly used to obtain the osteoclasts, and the receptor activator of nuclear factor κB ligand and macrophage colony-stimulating factor can be used in combination in order to obtain the mature osteoclasts. OBJECTIVE: To observe the biological characteristics of mouse monocyte macrophage RAW264.7, and to analyze the feasibility of receptor activator of nuclear factor κB ligand-mediated differentiation of monocyte macrophage RAW264.7 into osteoclasts. METHODS: The RAW264.7 cells were cultured, and then the RAW264.7 cells were induced with receptor activator of nuclear factor κB ligand for 7 days to observe the tartrate-resistant acid phosphatase staining results. The positive tartrate-resistant acid phosphatase staining and nucleus ≥ 3 were considered as the osteoclasts. Phalloidin staining was used to observe fibrous actin ring, and toluidine blue staining was used to observe the resorption pits on the surface of bovine bone slice.  RESULTS AND CONCLUSION: RAW264.7 cells could be induced into tartrate-resistant acid phosphatase-positive multinuclear cells with receptor activator of nuclear factor κB ligand, and formed fibrous actin rings; round or ellipse bone resorption pits were found in bone slice surface by microscope. RAW264.7 cells were considered as a good preosteoclast model that used for research of osteoclast differentiation. Receptor activator of nuclear factor κB ligand can induce the differentiation and maturation of RAW264.7 cells, reduce the application of macrophage colony-stimulating factor and make the training system simpler and easier to operate, and the induced cells have the high purity suitable for biological and biochemical studies of osteoclasts. Figures and Tables | References | Related Articles | Metrics

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Angiogenesis of dermal scaffold transplantation zone promoted by moboilizing endothelial progenitor cells Zhang Wei, Cai Di-xin, Wang Hong, Zhu Yong-cui 2013, 17 (24):  4386-4393.  doi: 10.3969/j.issn.2095-4344.2013.24.003 Abstract ( 493 )   PDF (2353KB) ( 387 )   Save BACKGROUND: In recent years, erythropoietin is found to mobilize autologous vascular endothelial progenitor cells from the bone marrow to peripheral blood, which are moved to the wound and promote wound angiogenesis and wound healing ultimately. OBJECTIVE: To observe the effect of erythropoietin on the mobilization of vascular endothelial progenitor cells.    METHODS: The vascular endothelial progenitor cells were mobilized by intraperitoneal injection of erythropoietin with different concentrations for 7 days, and the cells injected with normal saline were as control. The number of endothelial progenitor cells in the peripheral blood was detected with flow cytometry to compare the mobilization effect. The safe and effective erythropoietin mobilization dose was selected to observe the effect of erythropoietin- mobilized endothelial progenitor cells on the angiogenesis of the dermal scaffold transplantation zone.  RESULTS AND CONCLUSION: In 1, 3, 5, 7, 10 and 14 days after injection, there was no significant change in the number of endothelial progenitor cells in the peripheral blood of the normal saline group, while the number of endothelial progenitor cells in the peripheral blood of the erythropoietin mobilization group was increased gradually and reached peak at 7 days after injection. The proportion of endothelial progenitor cells was significantly increased in the high-dose erythropoietin mobilization group when compared with the low-dose erythropoietin mobilization group. The erythropoietin mobilization group had better effect on the angiogenesis in the dermal scaffold transplantation zone than the control group. Consecutive intraperitoneal injection of erythropoietin for 7 days can increase the number of endothelial progenitor cells in the peripheral blood that reaches peak at 7 days after mobilization. The mobilization effect is in a dose-dependent manner, and the injection can promote the angiogenesis in the dermal scaffold transplantation area. Figures and Tables | References | Related Articles | Metrics

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Estrogen effects on serum interleukin-8 and interleukin-10 expression in ovariectomized rats with osteoporosis Lu Shou-ming, Lu Shou-liang, Sun Tian-wei, Zhang Hang, Wang Qi-ming 2013, 17 (24):  4394-4400.  doi: 10.3969/j.issn.2095-4344.2013.24.004 Abstract ( 316 )   PDF (625KB) ( 436 )   Save BACKGROUND: The prevalence of osteoporosis is high in postmenopausal women, and whether estrogen prevents and treats osteoporosis through regulation of cytokine expression remains unclear. OBJECTIVE: To observe the effect of estrogen on the expression levels of the cytokines, interleukin-8 and interleukin-10, and to explore the correction between bone mineral density and cytokines. METHODS: Thirty healthy female Sprague-Dawley rats were included in this study and they were randomly divided into three groups: normal control group, model group and estradiol group. The rats of normal control group were subjected to laparotomy, while those of model group and estradiol group were completely ovariectomized. At 3 weeks after modeling, each rat of normal control group and model group was gavaged with 1.5 mL physiological saline every day, while the rats of estradiol group were given 1.5 mL estradiol valerate (1 mg/kg per day), for 12 weeks. The bone mineral density of femur and tibia was measured by dual-energy X-ray absorptiometry, and serum interleukin-8, interleukin-10 and estradiol levels were determined by enzyme linked immunosorbent assay. RESULTS AND CONCLUSION: The serum expression levels of estradiol in the model group and estradiol group were significantly decreased compared with the normal control group (P < 0.05), indicating the success of ovariectomy. The serum interleukin-8 level was significantly higher in the model group than the normal control group and estradiol group (P < 0.01), but serum interleukin-10 levels were significantly higher in the latter two groups (P < 0.01). There was no significant difference in the interleukin-10 expression level between normal control group and estradiol group (P > 0.05). The bone mineral density of femur and tibia was significantly negatively correlated with serum interleukin-8 level (P < 0 .001), and significantly positively correlated with the serum interleukin-10 level (P < 0.05). Estrogen could down-regulate the expression level of interleukin-8, and up-regulate the expression level of interleukin-10. By regulating the cytokine expression levels, estradiol may be another mechanism of action for the prevention and treatment of osteoporosis in postmenopausal women. Figures and Tables | References | Related Articles | Metrics

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Cardiac function and electrophysiological characteristics in myocardial infarction rats after tissue engineered cardiac patch transplantation Adila•Azhati, Zhao Long, Wang Qian, Ma Yi-tong 2013, 17 (24):  4401-4408.  doi: 10.3969/j.issn.2095-4344.2013.24.005 Abstract ( 330 )   PDF (941KB) ( 611 )   Save BACKGROUND: The structure of tissue engineered cardiac muscle tissue is similar to heart tissue three-dimensional electric coupling network and muscle striations, and has myocardial tissue-like systolic functions, and it can be used to repair the injured cardiac tissue.    OBJECTIVE: To observe the cardiac functions and electrophysiological characteristics in myocardial infarction rats after myocardial cells/collagen complex transplantation. METHODS: Adult Sprague-Dawley rats were randomly divided into three groups: sham surgery group, model group and transplantation group. Myocardial infarction models were developed in the model group and transplantation group, while sham surgery group was only subjected to thoracotomy with no ligation of coronary artery. Transplantation group was given myocardial cells/collagen complex transplantation, while sham surgery group and model group received no transplantation. RESULTS AND CONCLUSION: (1) Left ventricular heart function: Compared with the sham surgery group, the left ventricular internal diameter at end-diastole and left ventricular internal diameter at end-systole were significantly increased (P < 0.01), while left ventricular ejection fraction and shortening fraction were decreased in the model group (P < 0.01). The above indexes were not changed in transplantation group (P > 0.01). (2) Left ventricular effective refractory period: Compared with the sham surgery group, effective refractory period at the infarct border zone was significantly shortened in the model group (P < 0.01); compared with the model group, effective refractory period at the infarct border zone was significantly prolonged in transplantation group (P < 0.01). (3) Connexin 43 immunofluorescence results: Connexin 43 expression was positive in sham surgery group, weakly positive in the model group and transplantation group. However, the positive expression in the transplantation group was higher than that in model group. Myocardial cells/collagen complex transplantation can improve ventricular myocardium systolic function and electrophysiological properties of myocardial infarction rats, through electric coupling network and contraction coupling in tissue and structure. Figures and Tables | References | Related Articles | Metrics

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Left ventricular papillary muscle ablation in canines by ultrasound ablation catheter Wang Qi-feng, Wang Zhi-gang, Huang Jing 2013, 17 (24):  4409-4413.  doi: 10.3969/j.issn.2095-4344.2013.24.006 Abstract ( 468 )   PDF (1344KB) ( 405 )   Save BACKGROUND: Studies in recent years have found that the left ventricular papillary muscle plays an important role in ventricular arrhythmia. OBJECTIVE: To observe the feasibility and safety of homemade ultrasound ablation catheter in animal left ventricular papillary muscle ablation with transthoracic echocardiography.  METHODS: Nine ordinary mongrel dogs were collected and divided into two groups: control group (n=3) and ablation group (n=6). The carotid artery was separated with 3% pentobarbital sodium under sterile conditions, and implanted into the 8F arterial sheath was into to perform the mechanical ventilation, then the surface electrocardiogram was recorded. The abutting relationship between the catheter and papillary muscle was adjusted with transthoracic echocardiography image in order to keep the excellent contact between the catheter and papillary muscle. Then, 50 W/cm² acoustic intensity was applied to ablate the papillary muscle for 60 seconds. RESULTS AND CONCLUSION: All the animals were normally survived during the observation period, the anatomical structure of papillary muscle was clearly displayed on the transthoracic echocardiography image, and the relative position and abutting relationship between catheter and papillary muscle could be confirmed with the transthoracic echocardiography image. After cutting the animal myocardium, the ablation lesions in the papillary muscle could be seen, which was white in central and light red around and a clear boundary with the surrounding normal myocardium. The clear boundary between ablation lesions and the surrounding tissues could be seen under light microscope, and many necrotic cardiomyocytes were observed in the ablation lesion which space was increased and infiltrated with red blood cells. No associated complications of ablation (such as thromboembolism, myocardial burned or perforation and cardiac rupture) were detected. The catheter could achieve the ablation of deep papillary muscle effectively and safely by transthoracic echocardiography real-time monitoring. Figures and Tables | References | Related Articles | Metrics

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Simultaneous isolation of myocardial cells and cardiac fibroblasts from neonatal rats Shi Yu-lu, Li Xiao-yuan, Cao Mei-na, Yu Shu-yuan, Wang Ping 2013, 17 (24):  4414-4420.  doi: 10.3969/j.issn.2095-4344.2013.24.007 Abstract ( 621 )   PDF (1879KB) ( 1916 )   Save BACKGROUND: Myocardial cells and cardiac fibroblasts are simultaneously isolated through optimizing the culture conditions of myocardial cells, by use of the variations of adhesion properties of myocardial cells. OBJECTIVE: To explore the co-separation method for myocardial cells and cardiac fibroblasts in newborn rats. METHODS: Myocardial tissue in newborn rats was digested with low concentration trypsin at cold conditions and with collagenase II at 37 ℃. The myocardial cells and cardiac fibroblasts were isolated with differential centrifugation method, and myocardial cell culture conditions were improved using different kinds of serum. Immunofluorescence staining was used to evaluate the purity of myocardial cells and cardiac fibroblasts. RESULTS AND CONCLUSION: The myocardial Troponin T positive cell rate was 89.3% at culture 48 hours, and myocardial fibroblast vimentin positive cell rate was 93.6% at passage 2. The percentage of positive myocardial cells cultured with medium containing horse serum was obviously higher than that with bovine serum culture (P < 0.05). This isolation method of rat myocardial cells can gain myocardial cells and cardiac fibroblasts at the same time, with high purity and good activity. Figures and Tables | References | Related Articles | Metrics

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Gene expression of the skin and sweat glands in different development stages of embryo Chen Li-ying, Fan Kun-wu, Wang Jin-shui, Yu Xu-ming, Xu Cheng 2013, 17 (24):  4421-4428.  doi: 10.3969/j.issn.2095-4344.2013.24.008 Abstract ( 400 )   PDF (2556KB) ( 571 )   Save BACKGROUND: Because of the shortage of genetic evidence for the occurrence and development of the sweat glands as well as the relationship between sweat gland injury and regeneration, various reasons caused skin injury cannot reach complete repair of anatomical structure and physiological function. OBJECTIVE: To observe the morphology of skin tissue, and the occurrence and development pattern of the sweat gland, a skin appendage, in different development stages of embryo, as well as to find the related genes that play important roles in the development of sweat glands. METHODS: Thirteen fetuses with the gestational age of 12-36 weeks were provided by the Second People’s Hospital of Shenzhen, which strictly followed the ethical principles, and informed consent was written by their families. Hematoxylin-eosin staining and ABC immunohistochemical method were used to observe the structure and characteristics of fetal skin tissue and sweat glands in different development stages of embryo. Reverse transcription-PCR was used to analyze the gene expression of fetal skin tissue and sweat glands in the growth process. RESULTS AND CONCLUSION: Skin tissue structure of human fetus with different gestational ages showed regular change, which could be roughly divided into three development stages: the early formation of sweat glands (germ or the original base period, E12-15 weeks), the middle formation of sweat gland (E16-24 weeks), sweat gland maturation period (E25+-36 weeks). The keratin 7, keratin 8, keratin 18 and carcinoembryonic antigen biomolecular markers were positively expressed in the sweat gland cells of fetal skin tissue, while negatively expressed in the other cells of dermis and epidermis. The keratin 19 was not the specific marker of epidermal stem cells. Epidermal growth factor and its receptor are actively involved in the skin and sweat gland morphogenesis, which play an important role in maintaining the function of skin and sweat glands. Figures and Tables | References | Related Articles | Metrics

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Botulinum toxin type A induces apoptosis of human hypertrophic scar fibroblasts Li Wei-hua, Gao Yu-wei, Li De-shui, Xing Yu-xi 2013, 17 (24):  4429-4435.  doi: 10.3969/j.issn.2095-4344.2013.24.009 Abstract ( 383 )   PDF (1502KB) ( 448 )   Save BACKGROUND: Botulinum toxin type A can be used to treat human hypertrophic scar clinically and inhibit the proliferation of human hypertrophic scar fibroblasts in vitro. OBJECTIVE: To investigate the effect of botulinum toxin type A on the proliferation and apoptosis of human hypertrophic scar fibroblasts. METHODS: Human hypertrophic scar fibroblasts were isolated and cultured using digestion method. The cells were treated with botulinum type A at different concentrations. After 3-(4,5-dimethylthiazol-2-yl)-2,5- diphenyltetrazolium bromide dyeing, the cell solutions were detected with enzyme-linked immunosorbent detector at 570 nm. The inhibitory rate was calculated. The cultured cells were stained with hoechst 33342 and PI. The apoptotic cells were observed under fluorescence microscopy and the apoptotic rate was calculated. RESULTS AND CONCLUSION: The hypertrophic scar fibroblasts represented the spindle-like character with uniform composition and strong proliferation. After treated with botulinum toxin type A, the cells grew slowly, the cell amount reduced and the arrangement of the cells was scattered. The absorbance after 3-(4,5-dimethylthiazol- 2-yl)-2,5-diphenyltetrazolium bromide dyeing decreased. There was a statistically significant difference compared with the control group (P < 0.05). The half inhibitory rate was 0.4 IU/L. After hoechst 33342 and PI dyeing, the nuclei became blue and showed a smooth round or oval appearance. After treated with botulinum toxin type A, the nuclei were condensed with uneven dyeing. Some of the nuclei were fragmented and apoptotic bodies appeared. The apoptotic rate was increased with increasing concentrations of botulinum toxin type A. There was a statistically significant difference between the control group and the treated group (P < 0.05). The half apoptotic rate was 0.4 IU/L. These findings indicate that botulinum toxin type A can cause apoptosis of the human hypertrophic scar fibroblasts, thus inhibiting the proliferation of the cells. Figures and Tables | References | Related Articles | Metrics

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Notoginseng glycosides effects on hyperplastic scar Liu Kai, Pan Liang-liang, Li Ting, Liu Shu-fa 2013, 17 (24):  4436-4443.  doi: 10.3969/j.issn.2095-4344.2013.24.010 Abstract ( 529 )   PDF (2196KB) ( 563 )   Save BACKGROUND: Traditional treatments for hypertrophic scars, such as surgical resection, steroid hormones, antimetabolite, immunosuppressants and radiotherapy, are prone to resulting in relapse or serious adverse effects that limit their clinical application. In recent years, extraction of Chinese medicine for invigorating blood circulation and eliminating stasis has made satisfactory progress in the treatment of hypertrophic scars, and causes little adverse reactions. OBJECTIVE: To observe the effect of notoginseng glycosides on rabbit ear hyperplastic scar and mRNA expression of transforming growth factor beta 1 using histological staining and related gene detection methods. METHODS: A rabbit ear model of hyperplastic scar was established. At 4 weeks after modeling, 24 model rabbits were randomly divided into three groups, eight rabbits in each group: notoginseng glycosides group, triamcinolone acetonide group (positive control group) and blank control group. Local administration was performed in the three groups, five times. Then, scar tissues were extracted and dyed with Van-Gieson staining and hematoxylin-eosin staining for observation of collagen fibers and fibroblasts. Expression of transforming growth factor beta 1 mRNA was detected suing reverse transcription-PCR method. RESULTS AND CONCLUSION: The number of fibroblasts number in the scar was lower in the notoginseng glycosides group than the blank control group, but there was no difference between the triamcinolone acetonide group and notoginseng glycosides group. Collagen fibers arranged more regularly in the notoginseng glycosides group as compared with the blank control group, and there was also no difference between the triamcinolone acetonide group and notoginseng glycosides group. Expression of transforming growth factor beta 1 mRNA was significantly lower in the triamcinolone acetonide and notoginseng glycosides groups than the blank control group (P < 0.05). These findings indicate that notoginseng glycosides and triamcinolone acetonide can inhibit fibroblast over-proliferation and reduce the synthesis of collagenous fibers by reducing expression of transforming growth factor beta 1 mRNA, and thereby, the excessive hyperplasia of scar is inhibited. Figures and Tables | References | Related Articles | Metrics

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Two kinds of intermaxillary traction influence the mandibular micro-implant and its surrounding: Three-dimensional finite element analysis Zhao Gang, Wei Jia-jia, Zhang Xiao-ping, Yuan Xu, Shen Jian-xuan, Zhang Wei 2013, 17 (24):  4444-4450.  doi: 10.3969/j.issn.2095-4344.2013.24.011 Abstract ( 388 )   PDF (642KB) ( 509 )   Save Figures and Tables | References | Related Articles | Metrics

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Three-dimensional finite element analysis of the maxillary first premolar loaded with different biological forces Sun Hong-li, Yang Jian-jun, Xu Guo-hao, Gu Fang 2013, 17 (24):  4451-4456.  doi: 10.3969/j.issn.2095-4344.2013.24.012 Abstract ( 406 )   PDF (698KB) ( 553 )   Save BACKGROUND: The research on the micro-implant anchorage with finite element analysis method is rare, while accuracy or not of the finite element model is the main factor to determine the finite element analysis results. OBJECTIVE: To establish the three-dimensional finite element model of maxillary first premolar and to analyze the moving characteristics of maxillary first premolar under biological forces from different angles. METHODS: The data of the maxillary first premolar was obtained by dental CT scanning, and the conversion between CT image and three-dimensional model could be achieved by Mimics software. The three-dimensional finite element model was established by using the DICOM data modeling method, in order to simulate the biomechanical experiment of loading forces on the micro-implant anchorage from different angles, and then the moving characteristics of maxillary first premolar was analyzed. RESULTS AND CONCLUSION: The application of this method to obtain three-dimensional finite element model of maxillary first premolar was simple and accurate, and could simulate the mechanics experiment in the greatest degree. Maxillary first premolar produces compound motor trend of distal and inclined moving, distal and lingual rotating and down movement in action of micro-implant anchorage. Figures and Tables | References | Related Articles | Metrics

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Number of mast cells and expression of transforming growth factor beta 1 in the submandibular gland of diabetes mellitus rats Zhu Jian-hua, Tang Chun-ling, Zhang Yan-qiu, Liu Ji-guang 2013, 17 (24):  4457-4464.  doi: 10.3969/j.issn.2095-4344.2013.24.013 Abstract ( 362 )   PDF (2341KB) ( 462 )   Save BACKGROUND: Continuous hyperglycemia may result in degenerative changes in submandibular gland cells, showing the increased cytokine expression, altered glycogen metabolism and attenuated secretion function. Little evidence is available addressing the mast cells and transforming growth factor beta 1 in diabetic submandibular gland lesions.  OBJECTIVE: To observe the number and morphological changes of mast cells, as well as transforming growth factor beta 1 expression in the submandibular gland of diabetic rats. METHODS: Thirty-two male Sprague-Dawley rats were randomly divided into an experimental group and a control group. The body weight and blood glucose levels of all rats were measured at 4, 8, and 12 weeks after alloxan injection was given in the experimental group. Submandibular gland tissues were harvested for hematoxylin-eosin staining and toluidine blue staining to count mast cells. Transforming growth factor beta 1- positive cells were measured using SP immunohistochemical staining method. RESULTS AND CONCLUSION: (1) Mast cells were expressed in submandibular gland of both normal rats and diabetic rats, and the number of mast cells gradually increased as the disease duration prolonged and was significantly higher in the experimental group than in the control group (P < 0.01). (2) The transforming growth factor beta 1-positive cells were expressed in submandibular gland acinar cells, granular convoluted tubule and striated duct cells in both normal rats and diabetic rats, and the positive granules were localized in the cytoplasm. The number of positive cells gradually increased along with the duration of diabetes and was significantly higher in the experimental group than in the control group (P < 0.01). The mast cells and upregulated expression of transforming growth factor beta 1 in the submandibular gland of diabetic rats are positively related to the duration of diabetes mellitus Figures and Tables | References | Related Articles | Metrics

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Establishing a mouse model of radiation-induced thymus injury Ning Chang, Yu Chang-lin, Hu Kai-xun 2013, 17 (24):  4465-4472.  doi: 10.3969/j.issn.2095-4344.2013.24.014 Abstract ( 384 )   PDF (2149KB) ( 410 )   Save BACKGROUND: The thymus plays an important role in maintaining the immune function of the body. Damage and repair mechanisms of radiation-induced thymus injury require further studies, but there lacks an appropriate animal models.     OBJECTIVE: To establish a mouse model of radiation-induced thymus injury. METHODS: Totally 160 female BALB/C mice were randomly classified into four groups, 40 mice in each group: control group received sham-irradiation, and the other groups were irradiated with different doses of 60Co γ-rays, 6 Gy, 9 Gy and 12 Gy. After the irradiation, the body mass, food and water intake, the thymus index, and thymus pathology were detected at 1, 7, 14, 21, 28 days; esophageal, trachea and lung pathological changes were observed at 7 days; thymus and peripheral blood T subsets of the mice were measured at 14 days. RESULTS AND CONCLUSION: ALL three doses (6 Gy, 9 Gy, 12 Gy) could damage mouse thymus. After 6 Gy radiation, radiation-induced thymus injury was relatively mild and repaired within 1 week. Radioactive esophagus and trachea injury were found in mice exposed to 12 Gy radiation. Only exposed to 9 Gy, the thymus had a typical change and other organs were normal. Therefore, single mediastinal 60Co γ-ray irradiation at a dose of 9 Gy can successfully produce a mouse model of radioactive thymus injury. Figures and Tables | References | Related Articles | Metrics

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In vivo concentration gradient of basic fibroblast growth factor after coronary venous retrograde perfusion Zhen Lei, Wang Xiao, Miu Huang-tai, Qiao Shi-bin, Wu Xing-xin, Qiao Yan, Liu Bai-qiu, Liu Xin-min, Que Bin, Nie Shao-ping 2013, 17 (24):  4473-4480.  doi: 10.3969/j.issn.2095-4344.2013.24.015 Abstract ( 409 )   PDF (2259KB) ( 552 )   Save BACKGROUND: In vitro studies have demonstrated that the concentration gradient of basic fibroblast growth factor can promote the migration and proliferation of stem cells. However, it is unclear whether in vivo basic fibroblast growth factor concentration gradient can be established by coronary venous retrograde perfusion. OBJECTIVE: To evaluate the safety of coronary venous retrograde perfusion and establish the concentration gradient of basic fibroblast growth factor between coronary venous blood and target myocardium, and to investigate the time window of basic fibroblast growth factor concentration gradient after coronary venous retrograde perfusion. METHODS: The acute myocardial infarction models were established with ligating method, and 1 week later, coronary venous retrograde perfusion of basic fibroblast growth factor was performed. After perfusion, the time of balloon dilation was 0, 5, 10 and 15 minutes. The animals were sacrificed immediately after balloon inflation lifting, and then the enzyme-linked immunosorbent assay was used to measure the concentration of basic fibroblast growth factor in the plasma, farcted myocardium and homogenates of myocardium in the infarct border zone. The basic fibroblast growth factor concentration gradient between the coronary venous blood and infarcted myocardium and myocardium in the infarct border zone was in vivo evaluated.   RESULTS AND CONCLUSION: The success rate of coronary venous retrograde perfusion was 100%. No death, cardiac tamponade, or ventricular arrthythmia occurred. There was no significant difference in concentration of basic fibroblast growth factor between coronary venous blood and infarcted myocardium at 5 and 10 minutes after perfusion, while the concentration of basic fibroblast growth factor in the infarcted myocardium was higher than that in the coronary venous blood and myocardium in the infarct border zone. There was no significant difference in the concentration between two groups after balloon dilation for 15 minutes. The results indicate that after coronary venous retrograde perfusion of basic fibroblast growth factor, the time of balloon dilation was 5-10 minutes which can establish stable basic fibroblast growth factor concentration gradient between the coronary venous blood and infarcted myocardium, and the concentration is highest in the infarcted myocardium. The stem cell infusion in this time window can enhance its transitional vitality. Figures and Tables | References | Related Articles | Metrics

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低氧诱导因子1α基因敲除小鼠椎间盘退变与益气化瘀方的干预 Wang Jing, Dong Fang-fang, Li Xiao-feng, Xu Jin-hai, Shu Bing, Shi Qi, Wang Yong-jun, Zhou Chong-jian 2013, 17 (24):  4481-4487.  doi: 10.3969/j.issn.2095-4344.2013.24.016 Abstract ( 313 )   PDF (2458KB) ( 504 )   Save BACKGROUND: Hypoxia-inducible factor 1 α is highly correlated with normal physiology and pathology of intervertebral disc cartilage because it can maintain normal activities of cartilage in hypoxia. After knockout of hypoxia-inducible factor 1α, cartilage cannot maintain the hypoxia state, resulting in nutritional disturbance, and hypoxia/ischemia in chondrocytes. Consequently, cartilage degeneration occurs. OBJECTIVE: To observe the degeneration of intervertebral disc cartilage end-plate in hypoxia-inducible factor 1α gene knockout mice and investigate the effects of Chinese herbal compound Yiqihuayu Prescription on intervertebral disc cartilage end-plate degeneration. METHODS: We collected 2.5- (n=6) and 4.5-month-old (n=6) hypoxia-inducible factor 1α gene knockout mice and wild type hypoxia-inducible factor 1α+/+ control mice were obtained by interbreeding. The mice were sacrificed at 2.5 and 4.5 months old. Lumbar vertebra at L4-6 levels was harvested for Safranin O/fast green, hematoxylin-eosin, immunohistochemical staining. Another 12 hypoxia-inducible factor 1α gene knockout mice of 0.5 month old were randomly assigned to normal saline and Yiqihuayu Prescription groups. Following intragastrical administration for 2 months, lumbar vertebra at L4-6 levels was harvested for Safranin O/fast green, hematoxylin-eosin, immunohistochemical staining and analysis. RESULTS AND CONCLUSION: For hypoxia-inducible factor 1α-/- mice, mice at 2.5 months old developed aging related cartilage loss and bony tissue appearance, in addition to cartilage defects, uneven distribution of cells, and chondrocyte reduction. In addition, type II collagen and Sox-9 expression in intervertebral disc cartilage decreased, while type X collagen and matrix metalloproteinase 13 expression increased. At 4.5 months old, the cartilage injury was worsened, type II collagen and Sox-9 expression further reduced, and type X collagen and matrix metalloproteinase 13 expression further increased. Compared with normal saline group, Yiqihuayu Prescription reduced ossification and defect of endplate cartilage, the number of chondrocytes increased, and the distribution was more uniform as shown by Safranin O/fast green and hematoxylin-eosin staining. Moreover, Yiqihuayu Prescription increased type II collagen and Sox-9 expression, but decreased type X collagen and matrix metalloproteinase 13 protein expression. The results indicate that intervertebral disc cartilage degeneration occurred in hypoxia-inducible factor 1α gene knockout mice, which was progressed with aging of mice. Yiqihuayu Prescription can attenuate the degeneration of intervertebral disc cartilage in hypoxia-inducible factor 1α gene knockout mice. Figures and Tables | References | Related Articles | Metrics

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Expression and significance of stromal cell derived factor-1 in the intervertebral disk after lumbar disc degeneration Liu Gang, Ma Xin-long, Deng Shu-cai, Chen Si 2013, 17 (24):  4488-4494.  doi: 10.3969/j.issn.2095-4344.2013.24.017 Abstract ( 352 )   PDF (1511KB) ( 437 )   Save BACKGROUND: Exact pathogenesis of lumbar disc degeneration is not very clear. Inflammation is involved in the mechanism of lumbar disc degeneration. Stromal cell derived factor-1, which belongs to chemokine family, is related to inflammation. OBJECTIVE: To determine the expression of stromal cell derived factor-1 in the intervertebral disk of patients with lumbar disc degeneration, and then to analyze the association of the expression levels of stromal cell derived factor-1 with the severity of lumbar disc degeneration. METHODS: Eighty-four patients with lumbar disc degeneration (lumbar disc degeneration group) and 28 patients with disc burst fractures (control group) were enrolled in this study. The intervertebral disk tissues were collected after surgery. The protein level of stromal cell derived factor-1 was determined by enzyme linked immunosorbent assay. The grade of disc degeneration was determined according to Schneiderman’s classification. Association between the levels of stromal cell derived factor-1 and the Schneiderman’s grade was analyzed. RESULTS AND CONCLUSION: The stromal cell derived factor-1 levels in the intervertebral disk were significantly elevated in the lumbar disc degeneration group compared with the control group (P < 0.01). The levels of stromal cell derived factor-1 were significantly increased in lumbar disc degeneration patients with grade 4 compared with those with grades 2 and 3. Furthermore, lumbar disc degeneration patients with grade 3 showed significantly elevated stromal cell derived factor-1 levels compared with those with grade 2. Spearman correlation analysis indicated that stromal cell derived factor-1 levels were positively correlated with Schneiderman’s grades (r=0.412, P < 0.01). These findings indicate that the levels of stromal cell derived factor-1 are significantly elevated in the intervertebral disk of patients with lumbar disc degeneration, and positively associated with the severity of lumbar disc degeneration. Stromal cell derived factor-1 may be involved in the mechanism of lumbar disc degeneration. Figures and Tables | References | Related Articles | Metrics

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Transforming growth factor-beta 1 expression in the degenerative intervertebral disc Dai Qi-yu, Yang Ting-tong, Yu Fang-fang, Wang Quan-zhi, Wang Yuan, Zhang Xiao-shuang 2013, 17 (24):  4495-4501.  doi: 10.3969/j.issn.2095-4344.2013.24.018 Abstract ( 330 )   PDF (717KB) ( 504 )   Save BACKGROUND: Transforming growth factor-β1 can promote the proliferation and differentiation of intervertebral disc cells, and participate in the repair process after injury. However, whether transforming growth factor-β1 is involved in the intervertebral disc degeneration remains unclear. OBJECTIVE: To analyze the expression of transforming growth factor-β1 in the degenerative intervertebral disc tissues, and to study the correlation between transforming growth factor-β1 and intervertebral disc degeneration. METHODS: Normal intervertebral disc tissues collected from 30 cases and degenerative intervertebral disc tissues collected from 530 cases were researched by hematoxylin-eosin staining, western blot assay and reverse transcription-polymerase chain reaction. The degenerative intervertebral disc tissues were classified in histopathology, and transforming growth factor-β1 expression in different intervertebral disc degeneration tissues was analyzed and compared with that in normal tissues. RESULTS AND CONCLUSION: Pathological diagnose by hematoxylin-eosin staining showed that, degenerative intervertebral disc tissues were classified into 4 pathological grades. Western blot and reverse transcription-PCR showed positive expressions of transforming growth factor-β1 in both normal and degenerative intervertebral disc tissues, however, transforming growth factor-β1 expression levels increased as degenerative changes and showed significant differences between degenerative and normal tissues(P < 0.01). Overexpression of transforming growth factor-β1 is positively correlated with intervertebral disc degeneration in human. Figures and Tables | References | Related Articles | Metrics

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Reconstruction of quadriceps femoris by anatomizing the ventral roots of spinal nerves in rats with spinal cord injury Li Wei, Zhong Gui-bin 2013, 17 (24):  4502-4508.  doi: 10.3969/j.issn.2095-4344.2013.24.019 Abstract ( 353 )   PDF (822KB) ( 383 )   Save BACKGROUND: Reconstruction of motor function of limbs in spinal cord injury is significant for patients with paraplegia. OBJECTIVE: To recover the innervations and muscle contraction function of quadriceps femoris in spinal cord injured rats by constructing the neural pathway using the ventral roots of spinal nerves above injured   segment of spinal cord and lumbar nerve that dominates the quadriceps femoris. METHODS: L1 ventral root was anatomized to L3 ventral root from Sprague-Dawley rats. After 6 months, axonal regeneration was found from L1 ventral root to L3 ventral root through the grafting nerve, which may reconstruct muscle contraction function. After nerve suture for 6 months, spinal cord injury was made in L2 segment for neuroelectrophysiology examination to observe innervation of quadriceps femoris. RESULTS AND CONCLUSION: After semi-dissection of L2 segment, wave from quadriceps femoris was recorded in electromyogram meter when stimulating the grafting nerve. After injured the segment of L2, we also record wave from quadriceps femoris when stimulating L1 ventral root. Results showed that anastomosis of the L1 ventral roots of spinal nerves above injured segment of spinal cord and the ventral root below the injured spinal cord segment can reconstruct the reflex path of innervation of quadriceps femoris, and the reflex center of quadriceps femoris in the spinal cord moves up to L1 segment. Figures and Tables | References | Related Articles | Metrics

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Application of continuous thermodilution method in beagle models with pulmonary arterial hypertension Chen Dan-dan, Zhou Da-xin, Guan Li-hua, Chen Fa-dong, Dong Li-li, Qian Ju-ying, Ge Jun-bo 2013, 17 (24):  4509-4514.  doi: 10.3969/j.issn.2095-4344.2013.24.020 Abstract ( 378 )   PDF (789KB) ( 601 )   Save BACKGROUND: Previous methods of invasive blood pressure monitoring in small animals with pulmonary hypertension are mediated by pressure tracings of biological signals acquisition system using right heart  catheter method, however, due to the limitations of experimental instruments and animal size, pulmonary vascular resistance and cardiac output cannot be measured with pulmonary artery catheter method. OBJECTIVE: To measure cardiac output and pulmonary vascular resistance in beagle models with pulmonary hypertension induced by dehydromonocrotaline by using Swan-Ganz seven-lumen floating catheter and Vigilance monitor system with principles of continuous thermodilution method, and to explore the application value of continuous cardiac output method in pulmonary hypertension models. METHODS: Ten beagles were randomized into two groups: control group (right atrium was injected with dimethylformamide), and experimental group (right atrium was injected with dehydromonocrotaline to establish pulmonary hypertension models). Prior to the administration and 8 weeks after administration, the right atrial systolic pressure, right ventricular systolic pressure, pulmonary arterial systolic pressure, mean pulmonary arterial pressure, pulmonary capillary wedge pressure and cardiac output of beagles in two groups were measured using Swan-Ganz floating catheter and Vigilance monitor system. RESULTS AND CONCLUSION: In the experimental group, the pulmonary vascular resistance was significantly increased (P=0.00), while cardiac output was significantly decreased (P < 0.05) at 8 weeks after dehydromonocrotaline injection. Continuous thermodilution method was superior to conventional interrupted thermodilution method. We concluded that floating catheter and Vigilance monitor system based on the principles of continuous thermodilution method is an accurate, stable, and repeatable method for measuring pulmonary vascular resistance and cardiac output in beagle models with pulmonary hypertension induced by dehydromonocrotaline, with less trauma. Figures and Tables | References | Related Articles | Metrics

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A Wistar rat model of radiation-induced masseter injury Dong Gang, Zheng Jian-jin, Li Tao, Xu Xin, Lu Shu-lai 2013, 17 (24):  4515-4520.  doi: 10.3969/j.issn.2095-4344.2013.24.021 Abstract ( 356 )   PDF (878KB) ( 403 )   Save BACKGROUND: Construction of a stable animal model of radiation-induced masseter injury is significant for the research of maxillofacial region tumor. OBJECTIVE: To establish the animal model of Wistar rats with radiation-induced masseter injury after irradiated with the radiation dose of 40 Gy. METHODS: A total of 20 male adult Wistar rats were selected and randomly divided into radiation injury group and control group. Radiation injury group was radiated with linear accelerator at a dose of 40 Gy in the masseter tissue. After irradiated for 28 days, pathological changes in the masseter tissue of the radiation injury group and control group were observed by light microscope and electron microscope; the gene expression of transforming growth factor-β1 was detected by reverse transcription-PCR. RESULTS AND CONCLUSION: After irradiated for 28 days at the dose of 40 Gy, the irradiation injury of structural damage and reduced vascular density were observed in the masseter tissue (P < 0.01), and the gene expression of transforming growth factor-β1 was increased (P < 0.001). The result indicates that masseter tissue irradiated with linear accelerator at a dose of 40 Gy can be considered as the model of radiation-induced masseter injury. Figures and Tables | References | Related Articles | Metrics

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Germline degradation of a mouse model of familial amyotrophic lateral sclerosis when breeding Cai Bin, Fan Dong-sheng 2013, 17 (24):  4521-4528.  doi: 10.3969/j.issn.2095-4344.2013.24.022 Abstract ( 487 )   PDF (668KB) ( 512 )   Save BACKGROUND: SOD1-G93A transgenic mouse is a classical morbid animal model in the investigation of amyotrophic lateral sclerosis. However, there are few reports about the germline deterioration of this transgenic mouse model when breeding. OBJECTIVE: To investigate the germline deterioration of mouse model of familial amyotrophic lateral sclerosis when breeding. METHODS: The times of pregnancy, litter size among different generations and the ratios of male and positive of the filial generation were compared to analyze the change of fertility of amyotrophic lateral sclerosis mouse model of SOD1-G93A transgenic mouse when breeding. And the times of invasion and death were compared to analyze the change of pathogenesis of this transgenic mouse when breeding. RESULTS AND CONCLUSION: The times of pregnancy, per litter size and per mice were decreased significantly (P < 0.05) when comparing the third and sixth generation with the first generation, and moreover there were over half of the mouse of the sixth generation who can not inbreed. The ratios of male and positive mouse of the filial generation also decreased, but the difference was not significant. The time of invasion and death delayed significantly by 6.37 days (P=0.004) and 9.67 days (P=0.022) respectively when comparing the sixth generation with the first generation, and the difference was significant. As a result, familial amyotrophic lateral sclerosis mouse showed deteriorated germline when breeding such as the fertility decline and the delay of the time of invasion and death. Figures and Tables | References | Related Articles | Metrics

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Molecular mechanism of spinal cord injury detected using microarray technology Xi Yue 2013, 17 (24):  4529-4538.  doi: 10.3969/j.issn.2095-4344.2013.24.023 Abstract ( 508 )   PDF (960KB) ( 681 )   Save BACKGROUND: The molecular mechanisms of different biological processes after spinal cord injury are not clear. OBJECTIVE: To analyze the research advance of the gene expression change after spinal cord injury by using microarray technology. METHODS: Elsevier and Web of Knowledge databases were retrieved by computer with key words of “microarray, spinal cord injury, gene expression” to search papers about microarray and spinal cord injury published between January 1972 and November 2012. Related papers addressing the molecular mechanism of spinal cord injury by using microarray technology were selected. According to inclusion criteria, 56 literatures were selected in this study. Two Chinese writings were also consulted. RESULTS AND CONCLUSION: The change of gene expression from acute phase to chronic phase following spinal cord injury could be detected by using microarray technology which is established on molecular research, and then, the relevant signaling pathway and transcription factors could be found. This technology not only plays a guiding role in further molecular research, but also can be used to find the suitable therapeutic targets of spinal cord injury on genetic level. The research advance of pathophysiology and the gene expression after spinal cord injury by using microarray technology was reviewed in this paper. The technological innovation of experimental design and microarray detection, the improvement of data analysis methods were put forward. The power of microarrays in finding potential therapeutic targets was also evaluated. Figures and Tables | References | Related Articles | Metrics

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Effect and mechanism of nestin in the spinal cord development and injury repair process Wang Shi-yun, Zhang Ruo-yu, Liu Xue-hong 2013, 17 (24):  4539-4544.  doi: 10.3969/j.issn.2095-4344.2013.24.024 Abstract ( 241 )   PDF (559KB) ( 473 )   Save BACKGROUND: As a specific marker of neurons, expression activity of Nestin to a certain extent reflects the evolution law of spinal cord nerve cells’s proliferation, differentiation and migration. OBJECTIVE: To review the molecular structure, biological function, and expression and distribution law of Nestin in spinal cord development and damage repair, then to provide previous research foundation in the transplantation of neural stem cells after spinal cord injury. METHODS: VIP and Wanfang databases (1990/2012) were retrieved by computer with the key words of “spinal cord, Nestin” in Chinese. At the same time, PubMed, Springer and e-book (full text) databases (1990/2012) were retrieved with the key words of “spinal cord, Nestin” in English. Totally 343 articles were retrieved, and finally 35 articles were included. RESULTS AND CONCLUSION: As a specific marker of neural stem cells, Nestin at embryo stage shows a space-time expression that has a inverse relationship with the mature degree of neural precursor cells, namely with neural cell differentiation and growth continued to mature, Nestin expression strength would gradually abate, and even disappear. And in spinal cord injury pathological situations, Nestin expression strength and quantity in damaged tissue zone also reflect the repair ability of damaged spinal cord tissues. So during the development of spinal cord tissues, Nestin expression strength, quantity and distribution is now an important index for judging the development of spinal cord nerve cells, which can provide significant preliminary basis for research work for clinical treatment of spinal cord injury. Figures and Tables | References | Related Articles | Metrics

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Resistance training and protein supplement: Improvement of elderly muscle protein metabolism Wen Cai-xiong 2013, 17 (24):  4545-4552.  doi: 10.3969/j.issn.2095-4344.2013.24.025 Abstract ( 682 )   PDF (585KB) ( 699 )   Save BACKGROUND: Elderly muscle protein metabolism appears negative balance status, which is considered to be the main reason for elderly muscle loss and muscle function decline. In recent years, many studies have confirmed that resistance training and protein supplements can effectively improve the phenomenon of the negative balance of the elderly muscle protein metabolism, increase the elderly muscle mass, and improve muscle function. OBJECTIVE: To review the effect of the elderly resistance training, protein supplement and resistance training combined with protein supplements on muscle protein metabolism and muscle function, and to explore possible ways to improve the elderly muscle function and inhibit muscle loss. METHODS: The CNKI database (2005-01/2012-12) and PubMed database (2010-01/2012-12) were retrieved by the computer for the literatures addressing muscle protein metabolism and the effect of protein supplements and resistance training on muscle protein metabolism in the elderly. The key words were “protein metabolism, muscle cells, muscle loss, resistance training, protein supplement” in Chinese and “muscle cell, resistance training, sarcopenia, protein metabolism” in English. RESULTS AND CONCLUSION: A total of 81 literatures were screened out, 22 literatures that did not meet the standards were eliminated, and then finally 59 literatures were included for the analysis and review. The results show that the negative balance of elderly muscle protein metabolism is the direct reason to elderly muscle loss. Resistance training and leucine-rich protein supplement can effectively enhance the the elderly muscle protein anabolism level, and improve muscle function of the elderly. But the elderly muscle protein anabolic response capability against resistance training and protein supplements is much lower than young people. Figures and Tables | References | Related Articles | Metrics

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Research and application progress of visual fixation component separation Wei Wei, Shi Geng-hu, Li Yu-tang, Zhang Bing, Gao Chuang 2013, 17 (24):  4553-4560.  doi: 10.3969/j.issn.2095-4344.2013.24.026 Abstract ( 583 )   PDF (759KB) ( 505 )   Save BACKGROUND: Studies have shown the eye movement during visual fixation not only includes the noise signal from the nervous system, but also contains the information of cognitive processing. Fixation components and their functions attracted attention increasingly. OBJECTIVE: To review the research progress and its present situation of visual fixation component separation. METHODS: We retrieved the articles in PebMed database and Google Scholar with the computer using the key words of “visual fixation, fixational eye movement, tremor, drifts, micro saccades”. And we also made a reference to some corresponding monograph. The articles with repeated, unconcerned or obsolete contents were eliminated, and 64 articles were included in the final analysis. RESULTS AND CONCLUSION: Visual fixation is a phenomenon of human visual perception in which the individual is maintaining gaze in a stationary object. There are three types of eye movement during fixation: tremor, drifts and micro saccades, and they are vital for maintenance of visibility. Recently, discussions are focused on the contribution of these three eye movements on the visual perception, mainly including preventing fading and visual system correction. In addition, micro saccades can be sharply separated by computing the estimator of eye movement velocity thresholds. Further researches on the neurophysiological basis of fixational eye movements and the development of techniques, especially the recording of neural activity and the retinal stabilization methods, will deepen our understanding of the phenomenon of visual fixation.   Figures and Tables | References | Related Articles | Metrics