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06 May 2012, Volume 16 Issue 19 Previous Issue    Next Issue

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Effects of silencing annexin A1 gene on proliferation and osteogenic differentiation of rabbit bone marrow mesenchymal stem cells  Liang Bo-wei1, Pan Xin-yuan2, Zhao Jin-min1, Yin Guo-qian2, Hu Feng1 2012, 16 (19):  3421-3426.  doi: 10.3969/j.issn.1673-8225.2012.19.001 Abstract ( 316 )   PDF (456KB) ( 352 )   Save BACKGROUND: Annexin A1 (ANXA1) protein is involved in regulation of cell proliferation and differentiation. Steroid-induced osteonecrosis of femoral head is related to altered proliferation and osteogeneic differentiation of bone marrow mesenchymal stem cells (BMSCs). OBJECTIVE: To investigate the effects of silencing ANXA1 gene on proliferation and osteogenic differentiation of rabbit BMSCs. METHODS: New Zealand white rabbit BMSCs were cultured in vitro and divided into three groups. In the RNA interference group, BMSCs were transfected with lentiviral vectors carrying short hairpin RNA (LV-shANXA1) to down regulate the expression of ANXA1 gene. In the negative control group, BMSCs were transfected with lentiviral vectors carrying nonsense gene sequences (LV-shANXA1-NC). In the blank control group, there was no intervention. RESULTS AND CONCLUSION: The positive rate of CD44, CD105 and CD45 in BMSCs after purification was 97.2%, 95.8 % and 2.5%, respectively. The infection efficiency of lentivirus was 80% at multiplicity of infection (MOI) of 50 and 95% at MOI of 100 in BMSCs after selection by puromycin. Real-time PCR and western blot results showed that the silencing efficiency was above 72.4% at MOI of 50. After down regulating the expression of ANXA1 gene, significant growth inhibition could be found in BMSCs; the alkaline phosphatase positive rate and alkaline phosphatase activity decreased, and the alizarin red staining results showed negative reaction in the subsequent induction of osteogenic differentiation in the cells. The experimental results show that silencing ANXA1 gene reduces the proliferation and osteogenic differentiation capacity of BMSCs, which may be one of mechanisms underlying steroid-induced avascular necrosis of femoral head. Related Articles | Metrics

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Bone marrow mesenchymal stem cells increased expression of urokinase type plasminogen activator and promoted apoptosis of hepatic stellate cells in rats    Ning Lin, Jiang Hai-xing, Qin Shan-yu, Zhang Jun-hong, Yang Wen, Meng Yun-chao 2012, 16 (19):  3427-3432.  doi: 10.3969/j.issn.1673-8225.2012.19.002 Abstract ( 276 )   PDF (473KB) ( 416 )   Save BACKGROUND: Bone marrow mesenchymal stem cells (BMSCs) promote the apoptosis of hepatic stellate cells (HSCs) by paracrine and activation of hepatocyte growth factor (HGF).  OBJECTIVE: To investigate the effects of BMSC regulation to urokinase type plasminogen activator on activity of hepatocyte growth factor, and the mechanism by which BMSCs induce the apoptosis of hepatic stellate cells. METHODS: Five groups were divided randomly: Co-culture group, in which rat BMSCs and HSCs were co-cultured by upper and down double-layer co-culture system; HSCs group, in which HSCs were simply cultured; fibroblasts group; UK122 pretreated group: 6 hours before co-culture of BMSCs and HSCs, urokinase type plasminogen activator specific inhibitor UK122 was added; BMSCs blank control group. RESULTS AND CONCLUSION: mRNA expression of urokinase type plasminogen activator in the co-culture group was significantly higher than in the HSCs group (P < 0.01). Compared with HSC group, HSC proliferation was significantly inhibited in the co-culture group at 24 hours (P < 0.01) in a time-dependent manner. After co-culture, HGF protein expression and HSC apoptosis were significantly increased (P < 0.01). After UK122 intervention, HGF expression and HSC apoptosis were significantly decreased (P < 0.01). These findings suggest that BMSCs promote the secretion of urokinase type plasminogen activator, increase the expression of active HGF, and promote the apoptosis of HSCs. Related Articles | Metrics

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Estrogen promotes osteogenic differentiation of mouse bone marrow mesenchymal stem cells in a dose-dependent manner   Zhu Xiao-fei1, Wang Yang2, Jin Yan3, Jia Li-hui4 2012, 16 (19):  3433-3437.  doi: 10.3969/j.issn.1673-8225.2012.19.003 Abstract ( 279 )   PDF (396KB) ( 488 )   Save BACKGROUND: The pathogenesis of postmenopausal osteoporosis and estrogen deficiency is closely related. OBJECTIVE: To investigate the effects of different concentrations of estrogen on osteogenic differentiation of mouse bone marrow mesenchymal stem cells (BMSCs) and the relationship with miRNA-26a. METHODS: Mouse femoral and tibial bone marrow was taken and BMSCs were isolated and purified by the whole bone marrow adherence method. Estrogen at different concentrations (0, 10-10, 10-9, 10-8, 10-7, 10-6 mol/L) was used for osteogenic induction. RESULTS AND CONCLUSION: Estrogen did not greatly influence the proliferation of BMSCs, but it could significantly enhance the osteogenic differentiation capability of BMSCs. At the same time, estrogen, in particular at a concentration of 10-9 mol/L, could promote RUNX2, OCN mRNA and RUNCX2, SP7 protein expression in BMSCs. However, 10-9 mol/L estrogen promoted miRNA-26a mRNA expression at the weakest capability. These findings suggest that estrogen can promote osteogenic differentiation of BMSCs in a dose-dependent manner, and microRNA-26a possibly exerts effects during this process. Related Articles | Metrics

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Co-cultivation of bone marrow stromal cells with rabbit osteoblast-like cells in a Transwell chamber for osteogenic differentiation  Zhang Shu-ying1, Duan Da-bo2, Zhang Li2, Wang Zhi-ying1 2012, 16 (19):  3438-3441.  doi: 10.3969/j.issn.1673-8225.2012.19.004 Abstract ( 295 )   PDF (322KB) ( 450 )   Save BACKGROUND: Osteogenic differentiation is previously limited by culture of dexamethasone, growth factors or osteoblast-like cells with bone marrow mesenchymal stem cells at 1:1. OBJECTIVE: To investigate the feasibility of in vitro co-cultivation of osteoblast-ike cells and bone marrow stromal cells (BMSCs) and osteoblast differentiation of BMSCs by osteoblast-like cells. METHODS: Passage 3 rabbit osteoblast-like cells and passage 3 rabbit BMSCs were co-cultured in Transwell chamber, and osteoblast-like cells were inoculated into the lower part of culture plate. BMSCs were inoculated on Transwell inner membrane, serving as experimental group. BMSCs were inoculated into Transwell chamber alone with basal medium in the lower part, serving as control group. RESULTS AND CONCLUSION: In the experimental group, BMSCs differentiated toward osteoblasts, and alkaline phosphatase activity was significantly higher compared with the control group (P < 0.05). In the experimental group, BMSCs were positive for alizarin red staining, showing red calcified nodules; after RT-PCR, the expression of core-binding factor α1 was observed. In the control group, calcified nodules were not observed. These findings suggest that Transwell chamber can help to achieve the in vitro co-cultivation of osteoblast-like cells and BMSCs and the osteogenic differentiation of BMSCs. Related Articles | Metrics

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Effects of hypoxia on proliferation and migration of rat bone marrow mesenchymal stem cells Wang Li-wei, Zhao Yu, Huang Xu, Huang Wen 2012, 16 (19):  3442-3446.  doi: 10.3969/j.issn.1673-8225.2012.19.005 Abstract ( 272 )   PDF (525KB) ( 455 )   Save BACKGROUND: Studies have demonstrated that turnover of transplanted bone marrow mesenchymal stem cells (BMSCs) with poor differentiation potential in ischemic environment greatly influences curative effects. OBJECTIVE: To investigate the effects of hypoxia changes on proliferation and homing ability of rat BMSCs.  METHODS: Sprague-Dawley rat BMSCs were isolated and then cultured under atmosphere of 1% O2 (hypoxia group) and 20% O2 (normoxia group) respectively for 0-7 days. Cells for Transwell chamber migration were classified into four groups by oxygen concentration at culture and migration: 1%O2 culture and 1%O2 migration, 20%O2 culture and 1%O2 migration, 1%O2 culture and 20%O2 migration, 20%O2 culture and 20%O2 migration. Migrated cells were counted. Protein levels of integrin β1 and intercellular adhesion molecule-1 in BMSCs cultured under different concentrations of oxygen were detected by western blot. RESULTS AND CONCLUSION: Migration and proliferation abilities of BMSCs were enhanced under the atmosphere of 1%O2 and the abilities were weakened under the atmosphere of 20%O2 (P < 0.05). Integrin β1 expression in BMSCs was increased under the atmosphere of 1%O2 (P < 0.05). These findings suggest that 1% O2 enhances the proliferation and migration abilities of rat BMSCs after transplantation. Related Articles | Metrics

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Effects of vascular endothelial cells on runt-related transcription factor 2 expression and osteogenic differentiation of bone marrow mesenchymal stem cells in a co-cultivation system Yang Rui-nian1, Liu Liu1, Wang Fu-ke1, Zhao De-ping2 2012, 16 (19):  3447-3452.  doi: 10.3969/j.issn.1673-8225.2012.19.006 Abstract ( 329 )   PDF (566KB) ( 382 )   Save BACKGROUND: Vascular endothelial cells can promote osteogenic differentiation of bone marrow mesenchymal stem cells (BMSCs), which provide nutrition supports for osteogenic differentiation of stem cells. OBJECTIVE: To investigate the effects of human umbilical vein endothelial cells (HUVECs) on the morphology, growth, differentiation of BMSCs as well as the runt-related transcription factor 2 (Runx2) gene expression in a co-cultivation system. METHODS: In a co-cultivation system, passage 3 BMSCs and passage 3 HUVECs were cultured together and served as experimental group. Passage 3 BMSCs were simply cultured and served as control group. At 4, 6, 10 days of culture, BMSCs morphology, alkaline phosphatase activity and Runx2 gene expression were determined.  RESULTS AND CONCLUSION: Compared with control group, BMSCs morphology was diverse, and cell conjunction and fusion appeared, and osteogenic differentiation was obvious during anaphase in the experimental group. At each time point, there was no obvious change in alkaline phosphatase activity in the control group, and alkaline phosphatase activity was significantly higher in the experimental group than in the control group (P < 0.05). At 6 and 8 days, Runx2 gene expression in the experimental group was about 4 times that in the control group (P < 0.01). This suggests that in the co-cultivation system, intravenous endothelial cells can promote Runx2 expression in BMSCs and induce osteogenic differentiation of BMSCs. Related Articles | Metrics

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Effects of Rho kinase inhibitor Y-27632 on proliferation and cell cycle of rat bone marrow mesenchymal stem cells   Liu Xiao1, Wan Mei-rong2, Liu Xiao-yun2, Yan Xian-liang1, Zhang Zheng-zheng3, Liu Kai1, Xu Tie1 2012, 16 (19):  3453-3456.  doi: 10.3969/j.issn.1673-8225.2012.19.007 Abstract ( 322 )   PDF (371KB) ( 370 )   Save BACKGROUND: Rho kinase inhibition can regulate dynamic reorganization of cytoskeletal proteins, activate transcriptional factor, and stimulate cell proliferation and differentiation. OBJECTIVE: To explore the effects of Rho kinase inhibitor Y-27632 on proliferation and cell cycle of bone marrow mesenchymal stem cells (BMSCs). METHODS: Rat BMSCs were isolated using whole bone marrow culture method and purified using adherent culture method in vitro. After two passages, BMSCs were incubated in maintenance medium containing Y-27632 (10 µmol/L, a commonly used working concentration) in 96-well plates. A control group in which BMSCs were not treated was designated. RESULTS AND CONCLUSION: The growth curves of BMSCs for moderate inoculation density was typically S-shaped. After BMSCs were incubated with Rho kinase inhibitor Y-27632，the lag phase was found during the first 3 days, the growth was accelerated, and the growth curve was upward. Flow cytometry indicated that (80.640±5.516)% of the BMSCs were in G0/G1  phase of cell cycle, (13.397±2.511)% in S phase. After stimulated by Y27632, (61.723±8.829)% of the BMSCs were in G0/G1 phase, and (29.380±7.630) % were in S phase. The cell proliferation during S stage was significantly higher after stimulation by Y27632 (P < 0.05). Rho kinase inhibitor Y-27632 can significantly promote more BMSCs to enter the stage of DNA synthesis, and promote the division and proliferation of BMSCs. Related Articles | Metrics

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Effect of bone marrow-derived mesenchymal stem cell transplantation on the expression of MAG, OMgp in perihematomal tissue in rats with intracerebral hemorrhage Yan Yu, Wang Yan-lin, Song Bo, Zhang Hui-li, Qin Jie, Wei Chen-ming, Wang Hai-li, Xu Yu-ming 2012, 16 (19):  3457-3461.  doi: 10.3969/j.issn.1673-8225.2012.19.008 Abstract ( 318 )   PDF (446KB) ( 319 )   Save BACKGROUND: Axon regeneration of central nervous system after injury is hard, which is associated with the elevated expression of the axon growth inhibitors: Nogo-A, MAG, OMgp around the lesions. OBJECTIVE: To investigate the effects of intravenous administration of bone marrow-derived mesenchymal stem cells (BMSCs) on the expression of MAG and OMgp in perihematoma tissue in rats with intracerebral hemorrhage (ICH). METHODS: BMSCs were isolated and cultured using the whole bone marrow adherent method. A rat model of ICH was established by stereotaxical injection of collagenase IV into the striatum. Sprague-Dawley (SD) rats were randomly divided into sham-operated group, ICH group (model) and ICH plus BMSCs intervention group (BMSCs group). Rats in the BMSCs group were transfused with BMSCs (1×107/rat) via tail vein at 24 hours after surgery. On days 1, 3, 7, 14 after surgery, neurological function tests were performed among all three groups and animals were sacrificed at the corresponding time points for detecting the expression of MAG, OMgp by immunohistochemical method. RESULTS AND CONCLUSION: No obvious neurological function deficit was observed in sham-operated group on days 1, 3, 7, 14 after surgery. In the model group, the degree of neurological function deficit reached the peak on day 1 after surgery, lasted to day 3, recovery could be observed on day 7, and much more recovery could be seen on day 14. Compared to model group, the BMSCs group had significantly lower mean neurological function rating scores on days 3, 7,14 after surgery (P < 0.05). The mean expression levels of MAG, OMgp in neurons and gliocytes in perihematoma tissue in the model and BMSCs group were both significantly higher than that in the sham-operated group on days1, 3, 7,14 after surgery; meanwhile, the BMSCs group had significantly lower mean expression levels than model group on days 3, 7,14 after surgery (P﹤0.05). Transplantation of BMSCs could significantly promote the neurological function recovery of a rat model of ICH, which might be associated with its downregulation of MAG, OMgp in perihematoma tissue. Related Articles | Metrics

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Sodium butyrate induces rat bone marrow mesenchymal stem cells to differentiate into cardiomyocytes in vitro   Dong Liang, Lian Feng, Yang Wen-gang, Wang Yong-yi, Xue Song 2012, 16 (19):  3462-3466.  doi: 10.3969/j.issn.1673-8225.2012.19.009 Abstract ( 313 )   PDF (598KB) ( 419 )   Save BACKGROUND: Histone acetylation is one of mechanisms underlying induced cardiomyocyte differentiation of bone marrow mesenchymal stem cells (BMSCs). OBJECTIVE: To investigate the effects of sodium butyrate, a histone deacetylate inhibitor, on differentiation of rat BMSCs into cardiomyocytes in vitro.  METHODS: Rat bone marrow mesenchymal stem cells (rBM-MSCs) were isolated, cultured and identified. Passage 2 cells were induced to differentiate into cardiomyocytes by 1 mmol/L sodium butyrate. RESULTS AND CONLUSION: The isolated BMSCs were positive for CD90, but they were negative for CD31, CD45. After addition of sodium butyrate, cell proliferation ability and histone deacetylase activity were significantly decreased, but no obvious apoptosis was observed. Real-time PCR results showed that after addition of sodium butyrate, the expression level of myocardial genes including GATA-4, MEF-2c, β-MHC was significantly increased (P < 0.05). Western blot results showed that after induced by sodium butyrate, the expression level of cardiac-specific proteins including connexin 43 and cardiac troponin T was significantly increased. Troponin T expression detected by immunofluorescence was consistent with that detected by western blot. These findings suggest that sodium butyrate can effectively induce BMSCs differentiation into cardiomyocytes in vitro. The mechanism may be related to inhibition of histone deacetylase activity and increase in histone acetylation. References | Related Articles | Metrics

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Role of CD4+ and interferon-γ-inducible nitric oxide synthane-nitric oxide pathway in treatment of myasthenia gravis by transplantation of umbilical cord mesenchymal stem cells Liu Hong-yan, Guo Jing-ming, Wang Hai-yan, Ye Song, Ran Chang-li 2012, 16 (19):  3467-3470.  doi: 10.3969/j.issn.1673-8225.2012.19.010 Abstract ( 304 )   PDF (373KB) ( 312 )   Save BACKGROUND: Several studies have demonstrated that CD4+ and interferon-γ (INF-γ)-inducible nitric oxide synthane (iNOS)-nitric oxide (NO) pathway is closely related to occurrence of myasthenia gravis. OBJECTIVE: To investigate the mechanism underlying CD4+ and IFN-γ-iNOS-NO pathway in treatment of myasthenia gravis by transplantation of umbilical cord mesenchymal stem cells. METHODS: Rat models of myasthenia gravis were treated by intravenous transplantation of umbilical cord mesenchymal stem cells. At the same time, a control group was set. The expression of CD4+ on lymphocytes was tested by flow cytometry, the level of IFN-γ, iNOS and NO by ELISA, colorimetry and Griess, respectively. RESULTS AND CONCLUSION: At 1 week after transplantation, the expression of CD4+ in the transplantation group was significantly higher (P < 0.01), while the level of IFN-γ, iNOS and NO was significantly lower, than in the model group (P < 0.01). These findings suggest that transplantation of umbilical cord mesenchymal stem cells can up-regulate the expression of CD4+ in the lymphocytes in the transplantation group to adjust IFN-γ-iNOS-NO pathway, down-regulate NO level so as to alleviate immune injuries. Related Articles | Metrics

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Umbilical cord blood mesenchymal stem cells transplantation for the treatment of cerebral ischemia injury in rats Liu Hui-chun1, Zhang Pei-pei1, Wang Shi-min2, Yan Xiao-ling3, Wang Xin-ping2, Zhang Wen-zhi4, Tang Fan3 2012, 16 (19):  3471-3475.  doi: 10.3969/j.issn.1673-8225.2012.19.011 Abstract ( 353 )   PDF (385KB) ( 467 )   Save BACKGROUND: There is yet no medicine that is confirmed to improve ischemic stroke. The high self-renewal ability and multiplex differentiation potential make it possible for stem cells to rebuild the structure and function of the central nervous system (CNS). OBJECTIVE: To observe the effect and safety of umbilical cord blood mesenchymal stem cells transplantation for the treatment of ischemic stroke in rat models and to investigate its possible mechanism. METHODS：Human umbilical cord blood mesenchymal stem cells were isolated in vitro by using density gradient centrifugation combined with adherent screening and labeled by BrdU before transplantation. The male SD rats were used to establish the middle cerebral artery occlusion model and then the models were randomly divided into 3 groups. In the transplantation group, the umbilical cord blood mesenchymal stem cells were transplanted into corpus striatum on the injury side at 7 days after modeling; the PBS group was given the same dose of PBS and the rats in model group were without treatment. RESULTS AND CONCLUSION: The recovery of mNSS score of transplantation group was better than that of model group (P < 0.05), and the recovery of mNSS score in PBS group was slower than that in model group within 35 days (P < 0.05), and had no significant difference at the 56th day (P > 0.05). BrdU staining positive cells and Nestin, microtubule rapsyn 2, glial fibrillary acidic protein, Ⅷ factor and vascular endothelial growth factor immunohistochemical double staining positive cells could be seen in the center and surroundings of the injury area in transplantation group. It indicated that umbilical cord blood mesenchymal stem cells could be isolated in vitro, and could survive and differentiate in rat host brain and promote the recovery of the neural function of middle cerebral artery occlusion model rats. The mechanism might be related to the transplanted cells differentiating into neuron-like and neural glial-like cells, and secreting neurotrophic factors and promoting angiogenesis of host cells. Related Articles | Metrics

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Immune response after transplantation of human umbilical cord-derived mesenchymal stem cells in a mouse model of acute Alzheimer’s disease  Yu Cheng-cheng1,2,3,Tang Yong-yong2,3, Sheng Hong-xia2,3, Liu Gang4, Hu Zeng-yao4, Zhou Wen-xia4, Li Gong-jie5, Zhang Bin2,3, Chen Hu2,3 2012, 16 (19):  3476-3481.  doi: 10.3969/j.issn.1673-8225.2012.19.012 Abstract ( 289 )   PDF (459KB) ( 580 )   Save BACKGROUND: Studies have demonstrated that concentration of cytokines in the peripheral blood and cerebrospinal fluid of Alzheimer’s disease patients changes greatly compared with that in the healthy persons, which adds evidence to the conclusion that Alzheimer’s disease is followed by immune response. This suggests that inflammatory response possibly participates in the cascade reaction of neuropathy in Alzheimer’s disease. OBJECTIVE: To investigate the relationship between immune response and the level of each cytokine in the peripheral blood in a mouse model of Alzheimer’s disease after transplantation of human umbilical cord mesenchymal stem cells (hUCMSCs). METHODS: Passage 5 hUCMSCs at a density of 1×109/L were prepared for use. C57 mice were randomly divided into four groups. In the hUCMSCs group, lateral ventricle injection of β-amyloid 1-42 and caudal vein injection of 1 mL hUCMSCs were performed. In the model group, lateral ventricle injection of β-amyloid 1-42 and caudal vein injection of 1 mL physiological saline were performed. In the physiological saline group, Alzheimer’s disease was not induced, but equal amounts of physiological saline were administered via the caudal vein. In the normal group, Alzheimer’s disease was not induced, and no treatment was given. RESULTS AND CONCLUSION: Morris water maze test results showed that compared with normal group, escape latency was prolonged, and the time spent in passing through the platform for the first time was increased, while the number that mice crossed through the platform was decreased in the model group. Compared with the model group, the escape latency was slightly shortened, the time spent in passing through the platform for the first time was decreased, and the number that mice crossed through the platform was slightly, but not significantly, increased in the hUCMSCs group. Compared with the normal group, N-acetylaspartic acid was decreased and myo-inositol was increased at 4 days after Alzheimer’s disease induction, and interleukin 1β, tumor necrosis factor-α in the peripheral blood were increased, but the increase in interleukin 10 was not obvious at 7 days after Alzheimer’s disease induction in the model group. Compared with the model group, level of each cytokine was decreased at 7 days after hUCMSCs injection. These results suggest that hUCMSCs for treatment of Alzheimer’s disease is implemented by regulation of immune response. Related Articles | Metrics

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Protective effects of human umbilical cord mesenchymal stem cells on lung fibrosis in newborn rats  Tu Hui-ying1, Wu Ben-qing1, Chen Li1, Zhang Yi2, Chen Zi-sheng3 2012, 16 (19):  3482-3486.  doi: 10.3969/j.issn.1673-8225.2012.19.013 Abstract ( 297 )   PDF (441KB) ( 420 )   Save BACKGROUND: Human umbilical cord mesenchymal stem cells (hUMSCs) have become the hot spot of cytotherapy research about chronic lung disease, because of its capacity of self-prolifetion, differentiation and paracrine effect. OBJECTIVE: To study the protective effects of hUMSCs on lung fibrosis and normal lung growth in newborn rats. METHODS: Thirty-two 2-day-old newborn rats were randomly divided into four groups: PBS control group, hMSCs group, bleomycin group and bleomycin+hUMSCs group. Rats in the latter two groups were prepared into models of lung fibrosis by intraperitoneal injection of bleomycin, and two cell groups were injected with huMSCs at 7 days. RESULTS AND CONCLUSION: In the bleomycin group, the hydroxyproline content was the highest, and pulmonary fibrosis was the most severe compared with other three groups (P < 0.05). Compared with PBS control and hUMSCs groups, the radial alveolar count and transforming growth factor β1 expression in lung tissue homogenate were significantly lower, but the mRNA expression of vascular endothelial growth factor was significantly increased in the bleomycin and bleomycin+hUMSCs groups (both P < 0.05). After treatment, the above indices were significantly reversed (P < 0.05). It indicates that hUMSCs can increase the mRNA expression of vascular endothelial growth factor, reduce the level of transforming growth factor β1 mRNA and can produce a protective effect on bleomycin induced fibrosis. However, there is no obvious difference on hydroxyproline content after hUMSCs intraperitoneal injection, it confirms that hUMSCs have no effect on normal lung development.  Related Articles | Metrics

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Induced differentiation of umbilical cord blood mesenchymal stem cells into urothelial cells on fibrin clots Guang Qian-qian1, Han Yu-huan2, Chang Ji-wu3 2012, 16 (19):  3487-3491.  doi: 10.3969/j.issn.1673-8225.2012.19.014 Abstract ( 357 )   PDF (546KB) ( 358 )   Save BACKGROUND: Whether umbilical cord blood mesenchymal stem cells (UCB-MSCs) can be induced to differentiate into urothelial cells on fibrin clots remains poorly understood. OBJECTIVE: To investigate the possibility of UCB-MSCs differentiating into urothelial cells on fibrin clots. METHODS: In the experimental group, the UCB-MSCs on umbilical cord blood fibrin clot were co-cultured with fetal urothelial cells. In the control group, the UCB-MSCs on umbilical cord blood fibrin clot were cultured in culture medium of DMEM/F12. RESULTS AND CONCLUSION: In the experimental group, the UCB-MSCs gradually became larger and were polygonal. The cells turned over their ultrastructure to urothelial cells under transmission electron microscope and were positive for immunhistochemistry staining of anti-cytokeratins (AE1/AE3). In the control group, the UCB-MSCs morphology presented spindle-like arrangement and were negative for immunohistochemistry staining of anti-cytokeratins (AE1/AE3). The UCB-MSCs on fibrin clot have the potential to differentiate into urothelial cells. Related Articles | Metrics

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Adipose tissue derived stem cells increase the survival rate of fat grafts  Yang Bo, Shen Hong-liang, Xu Zhi-fei, Li Song, Chen Kun 2012, 16 (19):  3492-3495.  doi: 10.3969/j.issn.1673-8225.2012.19.015 Abstract ( 370 )   PDF (327KB) ( 498 )   Save BACKGROUND: The adipose tissue derived stem cells (ADSCs) can secrete cell factors to promote neovascularization following directional induction in vitro. OBJECTIVE: To estimate the effects of ADSCs on the survival rate of fat grafts following the mixed transplantation of ADSCs and free fat tissues. METHODS: ADSCs were transfected by recombinant adenovirus encoding green fluorescent protein (Ad-EGFP) gene with different multiplicity of infection (MOI) values. Ad-EGFP-transfected cells at the ideal MOI were divided into experimental group and control group. Mixed transplantation of ADSCs and fat tissues into the back of nude mice was performed in the experimental group, and the simple fat tissues transplantation was taken in the control group. RESULTS AND CONCLUSION: Green fluorescence could be seen in ADSCs at 24 hours after transfection, the expression intensity at 5 days was the strongest, and the EGFP was still expressed after passage. The ideal transfection rate was 95.3% at a MOI value of 50. The ADSCs in the experimental group scattered in the fat cells and interlobular septum. Some of the ADSCs could be differentiated into vascular endothelial cells. The wet mass and survival rate of fat grafts in the experimental group were improved compared with the control group (P = 0.001), but the fat graft fibrogenesis and damaged condition were decreased (P = 0.001). The transplantation of ADSCs can increase the survival rate of fat grafts through promoting the angiopoiesis of free fat tissues. Related Articles | Metrics

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Autologous adipose derived stem cells loaded onto human umbilical cord Wharton’s Jelly matrix-derived scaffold for repair of full-thickness articular cartilage defects of the knee in rabbits Han Shuang1, Lu Shi-bi2◇, Liu Qiang1, Zhang Li2, Huang Jing-xiang2, Wang Ai-yuan2, Liu Shu-yun2, Zhao Bin2, Xu Wen-jing2, 2012, 16 (19):  3496-3501.  doi: 10.3969/j.issn.1673-8225.2012.19.016 Abstract ( 415 )   PDF (779KB) ( 518 )   Save BACKGROUND: Umbilical cord Wharton jelly matrix is rich in hyaluronic acid, glycosaminoglycan and collagen, with similar components to natural chondrocyte extracellular matrix. Therefore, human umbilical cord-extracted Wharton jelly matrix is likely to become an ideal tissue engineering scaffold material of cartilage tissue.  OBJECTIVE: To evaluate the therapeutic effects of autologous adipose derived stem cells loaded onto human umbilical cord Wharton’s Jelly matrix-derived scaffold in repair of full-thickness articular cartilage defects of the knee in rabbits. METHODS: After induced differentiation into chondrocytes, rabbit auatologous adipose derived stem cells at a final concentration of 1010/L were compounded with human umbilical cord Wharton’s jelly matrix-derived scaffold and further cultured for 1 week to construct tissue-engineered cartilage. The full-thickness articular cartilage defects of the knee in rabbits were repaired by the tissue-engineered cartilage (experimental group). The therapeutic effects were compared between experimental group, scaffold group and blank group. At 3 months after surgery, gross observation, histological examination, glycosaminoglycan (GAG) and total collagen quantification and biomechanical tests were applied to analyze the results. RESULTS AND CONCLUSION: In the experimental group, most of defects were repaired by hyaline cartilage. In the scaffold group, most of defects were repaired by fibrous tissue. In the blank group, the defects were repaired with less fibrous tissue or not repaired. It is feasible to use adipose derived stem cells as the seed cells of cartilage tissue engineering. The constructed cartilage by tissue engineering can effectively repair knee articular cartilage defects and human umbilical cord Wharton’s jelly matrix-derived scaffold can be used as a good cartilage tissue engineering scaffold. Related Articles | Metrics

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Isolation and identification of rat renal papilla stem cells  Wen Jing, Cheng Qing-li, Ma Qiang, Qi Yun 2012, 16 (19):  3502-3506.  doi: 10.3969/j.issn.1673-8225.2012.19.017 Abstract ( 290 )   PDF (469KB) ( 405 )   Save BACKGROUND: There is little research about isolation of renal stem cells from the renal papillary in living donor and the characteristics of the cells. OBJECTIVE: To isolate and identify the renal papilla stem cells in vitro, and compare their biological characteristics to bone marrow mesenchymal stem cells. METHODS: Cells in the tip of the renal papilla in Sprague-Dawley rats were isolated, and bone marrow mesenchymal stem cells were from the bone marrow cavities of femur and tibia. The cells were seeded in culture flasks respectively. DMEM/F12 media containing 10% fetal bovine serum were added into the culture flasks for culture and passage in vitro. When 70%-80% confluence, 0.25% Trypsin were used to digest and passage the cells. The third-passage cells were used for adipogenic and osteogenic differentiation. RESULTS AND CONCLUSION: The renal papilla stem cells showed a rose-shaped and vortex-like growth and the cells were short spindle and polygons shaped. The growth curves of the cells were “S”-shape. The cells showed positive staining by oil red O after adipogenic differentiation, and orange calcium deposition could be seen in the cells by alizarin red staining after osteogenic differentiation. The flow cytometer analysis showed that CD29, CD44 and CD90 positively expressed in these cells, while CD45 negatively expressed in them. It is indicated that tissue stem cells containing the characteristics of mesenchymal stem cells exist in the renal papilla. Related Articles | Metrics

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Rat bone marrow mesenchymal stem cells differentiation into neuron-like cells in vitro Kuang Tao1, Wang Lei2, Song Wen3, Meng Dong-mei4 2012, 16 (19):  3507-3510.  doi: 10.3969/j.issn.1673-8225.2012.19.018 Abstract ( 251 )   PDF (271KB) ( 305 )   Save BACKGROUND: Bone marrow mesenchymal stem cells (BMSCs) can be induced to differentiate into neuron-like cells in new environment after transplantation, and then to replace damaged cells for reconstructing neural circuit. OBJECTIVE: To establish the co-culture system between rat BMSCs and neural cells in vitro, and to study the influence of neural cells on the differentiation of BMSCs into neuron-like cells in the co-culture system. METHODS: The neural cells obtained from Wistar rat fetal brain tissue and BMSCs gained from rat thighbone were co-cultured in Transwell culture plate. The morphological changes of BMSCs were observed and the special markers of neural cells in BMSCs were examined by immunofluorescence on the fifth day of the co-culture. The results were compared with control group which where BMSCs were alone cultured. RESULTS AND CONCLUSION: BMSCs in the neural cells co-culture system extended, were radial, connected each other. Neuron specific enolase immunoreactions showed positive results, showing neuron-like cells. The positive ratio of neuron specific enolase-positive cells was (33.0±10.5)%. However, BMSCs in the control group did not express neuron morphological character. Immunofluorescence exhibited that cells were negative for neuron specific enolase. These indicate that microenvironment provided by neurons improves the differentiation of BMSCs into neuron-like cells. Related Articles | Metrics

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Preventive and therapeutic effects of intra-bone marrow cavity injection of bone mesenchymal stem cells on osteoporosis in rats subjected to simulated weightlessness Wu Li-feng1, Sun Ping2, Mai Yan-xing1, Liu Hai-ming1, Xu Yan-hua1, Yang Rui1, Huang Zhen3 2012, 16 (19):  3511-3514.  doi: 10.3969/j.issn.1673-8225.2012.19.019 Abstract ( 297 )   PDF (345KB) ( 434 )   Save BACKGROUND: Simulated weightlessness can inhibit the proliferation of bone marrow mesenchymal stem cells and their differentiation towards osteoblasts, which leads to osteoporosis characterized by low bone mass and microarchitectural deterioration of bone tissue. OBJECTIVE: To study the effects of intra-bone marrow cavity injection of allogeneic bone mesenchymal stem cells (BMSCs) on the bone mineral density and microarchitecture of the tibias in rats subjected to simulated weightlessness. METHODS: Male SD rats were randomly divided into three groups: normal control group, tail-suspended simulated weightlessness group, and treatment group. The bilateral tibias were subjected to intra-bone marrow cavity injection of allogeneic BMSCs while giving the tail-suspended simulated weightlessness. RESULTS AND CONCLUSION: Compared with the normal control group, the bone mineral density, percentage of trabecular area, trabecular number and trabecular thickness were decreased in the tail-suspended simulated weightlessness group (P < 0.01), while the trabecular separation was increased significantly (P < 0.01). Compared with the tail-suspended simulated weightlessness group, the bone mineral density, percentage of trabecular area, trabecular number and trabecular thickness were increased in the treatment group (P < 0.01), while the trabecular separation was decreased obviously (P < 0.01). The intra-bone marrow cavity injection can increase the bone mineral density of rats subjected to simulated weightlessness, improve the bone ultrastructure, slow down bone loss, and have beneficial preventive and therapeutic effects on osteoporosis..   Related Articles | Metrics

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Isolation, culture, and immune-phenotype identification of human synovium-derived mesenchymal stem cells and their immunosuppressive effects in mixed lymphocyte reaction system  Zhang Zheng-zheng, Li Wei-ping, Yang Rui, Song Bin, Wang Li-hui 2012, 16 (19):  3515-3519.  doi: 10.3969/j.issn.1673-8225.2012.19.020 Abstract ( 472 )   PDF (394KB) ( 564 )   Save BACKGROUND: There are mesenchymal stem cells (MSCs) in synovium that have the ability to proliferate extensively in culture. OBJECTIVE: To study the method of isolating and culturing human synovium-derived mesenchymal stem cells (SMSCs), to explore the immune-phenotype and the effect of SMSCs on the proliferation of T lymphocytes, and to evaluate the immunological  characteristics of SMSCs. METHODS: Cell populations were enzymatically released from the synovial membrane obtained from human knee joints by arthroscopy. Monoclone was selected as primary SMSCs. SMSCs were assayed on its multiplication capacity and cell vitality and characterized by FACS analysis. According to different ratios of SMSCs to peripheral blood monocytes, SMSCs were treated with mitomycin in the MLR system. The lymphocyte proliferation rate was measured by MTT assay and then the inhibition rate was calculated. RESULTS AND CONCLUSION: There was no statistical significance between the 10 clones in multiplication capacity and cell vitality (P > 0.05). SMSCs were positive for CD44, CD90, CD105, but negative for CD14, CD34, CD45 and HLA-DR, respectively. After the second passage, CD166 appeared to be positive. The proliferation of T lymphocytes could be inhibited to different extents by adding different ratios of SMSCs to human PBMCs, and the inhibition rate increased as the ratios of SMSCs to PBMCs increased. Human SMSCs can be isolated from knee joint. These cells have the ability to proliferate extensively in culture, express the phenotype of MSCs and exhibit immune regulatory effects. Related Articles | Metrics

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Feasibility of chlormethyl-benzamidodialkylcarbocyanine labeling rat kidney fat capsule adipose-derived mesenchymal stem cells  Zi Li-ying1, Zheng Hong-guang2, Zhang De-wei2, Mei Yu-ming2 2012, 16 (19):  3520-3524.  doi: 10.3969/j.issn.1673-8225.2012.19.021 Abstract ( 288 )   PDF (412KB) ( 347 )   Save BACKGROUND: Chlormethyl-benzamidodialkylcarbocyanine (CM-DiI) has been considered a relatively stable lipophilic fluorescent dye. OBJECTIVE: To investigate the labeling effects of CM-Dil for kidney fat capsule adipose-derived mesenchymal stem cells and its effects on cell morphology, growth and proliferation. METHODS: Sprague-Dawley rat kidney fat capsule adipose-derived mesenchymal stem cells were isolated by collagenase digestion. Passage 3 kidney fat capsule adipose-derived mesenchymal stem cells were labeled by CM-Dil. RESULTS AND RESULTS: Under fluorescent microscope, CM-Dil-labeled kidney fat capsule adipose-derived mesenchymal stem cells exhibited red fluorescence, with a labeling rate of over 95%. There were no obvious changes in cell morphology, growth and proliferation before and after labeling. After several passages, the number and staining intensity of cells with red fluorescence were decreased. At 15 days after CM-Dil labeling, over 50% of passaged cells were labeled. These suggest that CM-Dil can effectively label fat capsule adipose-derived mesenchymal stem cells and this method is easy-to-operate, with a high labeling rate, produces no cytotoxicitry and does not influence cell biological characteristics.  Related Articles | Metrics

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Insulin-like growth factor 1 promotes axon growth in neural stem cell-derived neurons Yue Ying-jie1, Fei Chang2, Zhang Jian2 2012, 16 (19):  3525-3528.  doi: 10.3969/j.issn.1673-8225.2012.19.022 Abstract ( 279 )   PDF (289KB) ( 407 )   Save BACKGROUND: Previous studies have shown that insulin-like growth factor 1 has a neuroprotective effect and can promote the differentiation of neural stem cells into neurons and oligodendrocytes. OBJECTIVE: To investigate the effects of insulin-like growth factor 1 on axon growth in the neural stem cell-derived neurons. METHODS: Hippocampal neural stem cells of newborn wistar rats were cultured and passaged. Passage 3-5 neural stem cells were incubated in a 24-well plate. 10 μL insulin-like growth factors (500 mg/L) were added into 12 wells (experimental group). The remaining wells served as control group. RESULTS AND CONCLUSION: At 1, 2, 3, 4 days of culture, dead cells in the experimental group were significantly fewer than those in the control group (P < 0.05). The length of neuronal axon was significantly longer in the experimental group than in the control group (P < 0.05), but there was no significant difference in bifurcation numbers between these two groups (P > 0.05). These findings suggest that insulin-like growth factor 1 can increase the differentiation of neural stem cells into neurons and promote the extension of neural stem cells-derived neuronal axon, but it cannot increase the number of axon bifurcations.   Related Articles | Metrics

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Matrix metalloproteinase-2 and matrix metalloproteinase-9 mRNA expression in mice with collagen-induced arthritis following heterogenous umbilical cord blood stem cell transplantation*☆ Niu Guang-hua, Gao Yu-jie, Du Jing, Guo He, Wang Bai-shan, Gao Ming-li 2012, 16 (19):  3529-3534.  doi: 10.3969/j.issn.1673-8225.2012.19.023 Abstract ( 329 )   PDF (375KB) ( 442 )   Save BACKGROUND: Matrix metalloproteinase (MMP) degrades extracellular matrix, which is a necessity of joint destruction in rheumatoid arthritis patients. MMP-2 and MMP-9 can evaluate rheumatoid arthritis and serve as an index to predict progressive destruction of the joint. OBJECTIVE: To observe heterogenous allogeneic umbilical cord blood stem cell (UBSC) transplantation on MMP-2 and MMP-9 expression in the spleen of mice with type Ⅱ collagen-induced arthritis. METHODS: Fetus cord blood was sterilely obtained and cord blood stem cells were separated. The C57BL/6(H-2b) mice were assigned to five groups (n=10). Except normal control group, models of collagen-induced arthritis were established using complete Freund’s adjuvant + type Ⅱ collagen. Mice from the methopterin group were intragastrically administered methopterin suspension 0.017 5 g/kg, once every 5 days. Other groups used caudal vein injection. Mice from the model and normal control groups were injected with saline. Mice from the mono-UBSCs group and double-UBSCs group were injected with 2×106/kg UBSCs from one and two parents. At 42 days following injection, animals were sacrificed and the ankle joint was obtained for histopathological detection. MMP-2 and MMP-9 mRNA expression in the spleen was examined using reverse transcription-polymerase chain reaction. RESULTS AND CONCLUSION: Double-UBSC transplantation could significantly inhibit inflammatory cell infiltration in synovial tissue of mice with type Ⅱ collagen-induced arthritis, repaired impaired cartilage tissue. The repair effect was better than that in methopterin group and mono-UBSCs group. MMP-2 and MMP-9 mRNA expression in the spleen was significantly lower in the double-UBSCs group than the mono-UBSCs group (P < 0.01). These suggest that heterogenous allogeneic double-UBSCs transplantation participated in pathological changes in rheumatoid arthritis cartilage and in synthesis of cartilage extracellular matrix and effectively treated rheumatoid arthritis by regulating MMP-2 and MMP-9 mRNA expression. Related Articles | Metrics

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Transplantation of allogeneic peripheral blood hematopoietic stem cells for treatment of leukemia Chen Yu-qing1, Li Lin2, Wang Long-an2, Shi Jie1, Zang Yu-zhu1, Zhang Yin1 2012, 16 (19):  3535-3539.  doi: 10.3969/j.issn.1673-8225.2012.19.024 Abstract ( 360 )   PDF (280KB) ( 447 )   Save BACKGROUND: Allogeneic peripheral blood hematopoietic stem cell transplantation is an effective method for treatment of leukemia. OBJECTIVE: To evaluate haemopoietic reconstitution, immune reconstitution, infection, incidence of graft-versus-host disease (GVHD) and other complications between related transplantation group and unrelated donor transplantation group. METHODS: Forty-five patients received allogeneic peripheral blood hematopoietic stem cell transplantation for treatment of leukemia. Clinical results of 30 patients receiving related transplantation were compared to those of 15 patients receiving unrelated donor transplantation. RESULTS AND CONCLUSION: The related transplantation group resulted in a significantly shorter time to neutrophil and platelet engraftment compared to the unre-lated donor transplantation group (P < 0.05). At 30-40 days after transplantation, en-graftment test revealed that allogeneic hematopoietic stem cells were implanted successfully into recipients. T cell reconstitution between the two groups differed slightly without significant difference at different time after transplantation. There were no significant differences in incidence of early-stage infection, acute and chronic GVHD between related donor transplantation group and unrelated donor transplantation group (P > 0.05). There were no significant differences in relapse of leukemia (P > 0.05) and 2-year disease free survival between related donor trans-plantation group and unrelated donor transplantation group (P > 0.05). These findings suggest that haemopoietic reconstruction in the related donor transplantation group was more rapid than in the unrelated donor transplantation group, but there were no significant differences in T cell reconstruction, incidence of infection and GVHD, and 2-year disease free survival rate between related donor transplantation group and unrelated donor transplantation group. Related Articles | Metrics

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Bone marrow stem cells transplantation for treatment of delayed encephalopathy after acute carbon monoxide poisoning in rats Xing Hong-xia1, 2, Yin Chuang3, Liu Sheng4, Zhu Yuan-kai2, Shi Li-jin2, Tian Xiao-jun2, Zhang Bo-ai1, Wang Yu-mei2 2012, 16 (19):  3540-3544.  doi: 10.3969/j.issn.1673-8225.2012.19.025 Abstract ( 318 )   PDF (438KB) ( 371 )   Save BACKGROUND: The therapy of delayed encephalopathy after acute carbon monoxide poisoning (DEACMP) in our country mainly depends on drug or hyperbaric oxygen. But the treatment has the characters of longer course, lower effect and higher cost. OBJECTIVE: To observe the changes of mature neurons and cell apoptosis before and after bone marrow mesenchymal stem cells (BMSCs) transplantation for treatment of DEACMP. METHODS: SD rats were randomly divided into three groups: sham operation group, control group and transplantation group. The rat model with DEACMP was established and intracerebral injection of allogeneic BMSCs was performed in the transplantation group at 0, 3, 5, 12, 24, 72 hours and 1 week after modeling, and cell transplantation was not performed in the control group. In the sham operation group, the external carotid artery was tied off and the PBS was injected instead of cell suspension.  RESULTS AND CONCLUSION: The expression of neuron-core protein (NeuN) in rat brain in the transplantation group at 3, 6,12 and 24 hours were significantly higher than that in the control group and sham operation group (P < 0.01). There was no significant difference in NeuN expression at 3, 6, 12 and 24 hours (P > 0.05). The average absorbance value of apoptosis in the transplantation group across 0-hour, 3-hour and 6-hour three time points was significantly lower than that in the sham operation group and other transplantation groups (P < 0.05). The average absorbance value of apoptosis in 1 week group was significantly lower than that in 2-week, 3-week and 4-week groups (P < 0.05). Transplantating BMSCs into the DEACMP rat models through the carotid artery could increase the quantity of mature neurons, and the underlying mechanism has relationship with apoptosis reduction. The best time for transplantation was between the 0 hour and the 24 hours after falling ill and the positive effect was significant at 1 week after transplantation. Related Articles | Metrics

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Intra-arterial transplantation of autologous bone marrow mesenchymal stem cells for treatment of diabetes in dogs    Wang Li-mei1, Cui Xiao-lan2, Ding Ming-chao1, Dou Li-dong1, Li Qian-qian1, Wang Yi-zhong1 2012, 16 (19):  3545-3550.  doi: 10.3969/j.issn.1673-8225.2012.19.026 Abstract ( 259 )   PDF (887KB) ( 348 )   Save BACKGROUND: At present, most researchers focus on intravenous transplantation of bone marrow mesenchymal stem cells (BMSCs) into diabetic animal models. There are few studies that describe transplantation of BMSCs into the pancreas of diabetic animal models via arterial intervention. OBJECTIVE: To investigate the distribution and differentiation of autologous BMSCs transplanted into the pancreas of a dog model of diabetes via the arterial intervention as well as the therapeutic effects and safety. METHODS: 30 dogs were randomly divided into BMSCs group (treatment group, n=13), diabetic model control group (model control group, n=10) and normal control group (n=7). Dogs in treatment group and model control group were intravenously injected with alloxan to establish diabetes models. After model establishment, treatment group received insulin therapy and autologous transplantation of BMSCs via arterial intervention. Model control group only received insulin therapy, and normal control group did not receive any treatment. RESULTS AND CONCLUSION: Compared with the model control group, insulin dosage significantly decreased and serum C-peptide level significantly increased (P < 0.05) in the treatment group (P < 0.05) at 12 weeks after transplantation. At 4 and   12 weeks after transplantation, the frozen sections of treatment group showed that the heart, liver, spleen, lung and kidney had clear structure, without necrosis and fibrosis. Compared with model control group and normal control group, the morphological structure of each organ had no obvious abnormal change after transplantation in the treatment group. At 4 weeks after transplantation, BMSCs mainly distributed in the pancreas and kidney, and CM-DiI and insulin double-positive cells were in the pancreas as detected by immunofluorescence staining. The results suggest that the method of transplantation of autologous BMSCs via arterial intervention for treatment of diabetic dogs is safe and effective.   Related Articles | Metrics

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RNA interference inhibits caspase-3 gene expression in bone marrow mesenchymal stem cells    Liu Li-bao1, Hua Ping1, Yang Song-ran2, Liu Jia-liang1, Jiang Hui-qi1, Wang Meng1, Zeng Kuan1, Ao Xiang1, Yang Yan-qi1 2012, 16 (19):  3551-3555.  doi: 10.3969/j.issn.1673-8225.2012.19.027 Abstract ( 280 )   PDF (426KB) ( 450 )   Save BACKGROUND: Poor viability of bone mesenchymal stem cells after transplantation into the infarcted region has limited the reparative capacity of these cells because of the apoptosis. OBJECTIVE: To construct eukaryotic vector expressing small interfering RNAs against caspase-3 and investigate the inhibitory effect on the expression of caspase-3 in BMSCs in vitro. METHODS: According to the principle of siRNA design, siRNAs against caspase-3 gene were designed and synthesized. Eukaryotic vector expressing caspase-3siRNA was constructed and transferred into BMSCs with the liposome conduct. There were 5 groups, including normal cell group, negative sequence group and 3 interfering groups (si-caspase-3-1, si-caspase-3-2 and si-caspase-3-3). RESULTS AND CONCLUSION: Eukaryotic vector expressing caspase-3siRNA was constructed and transferred into BMSCs successfully. Western blot analysis and reverse-transcription polymerase chain reaction demonstrated that levels of of caspase-3 protein and mRNA in transfected BMSCs decreased, with the most significant decline in the group with si-caspase-3-3. Eukaryotic vector expressing caspase-3 siRNA could inhibit the expression of caspase-3 gene in BMSCs in vitro. Related Articles | Metrics

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Effects of ginsenoside Rg3 on proliferation and induced differentiation of erythroleukemia cell line K562 in vitro  Xiao Feng1, 2, Liu Bin1, Zhang Jian-ping2, Wang Ban-qing2, Fang Mu-shui2, Hu Wei2, Sun Xu-Lu2, Zhu Qing-Xian1 2012, 16 (19):  3556-3559.  doi: 10.3969/j.issn.1673-8225.2012.19.028 Abstract ( 266 )   PDF (328KB) ( 388 )   Save BACKGROUND: Evidence exists that ginsenoside Rg3 can inhibit the growth of carcinoma cells. However, there are fewer studies describing ginsenoside Rg3 effects on leukemia. OBJECTIVE: To investigate the effects of ginsenoside Rg3 on proliferation of erythroleukemia cell line K562 and the underlying mechanism. METHODS: Erythroleukemia cell line K562 was used as target cell. The cells were divided into a control group and an Rg3 group. 10, 20, 40, 80, 100 mg/L Rg3 was added in the Rg3 group. RESULTS AND CONCLUSION: The MTT colorimetric assay indicated that K562 growth inhibition rate was significantly higher in the Rg3 groups than in the control group (P < 0.05-0.01). The nitroblue tetrazolium assay showed that after culture for 1, 2, 3 days, the reducing power of K562 cells in the Rg3 group was significantly higher than in the control group (P < 0.01). In the Rg3 group, some K563 cell somas became small, nucleoli disappeared, nitroblue tetrazolium positive cells increased, and cells developed toward maturation. Flow cytometry detection showed that after 2 days of culture, the cell cycle at G2 phase in Rg3 group was 3.84 times higher than that in the control group. Ginsenoside Rg3 can inhibit the proliferation of erythroleukemia cell line K562 in vitro. The underlying mechanism may be related to a fact that ginsenoside Rg3 blocks K562 cell cycle at G2 stage, hinders normal mitosis and thereby inhibits cell proliferation. Related Articles | Metrics

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Stem cells in the treatment of stroke  Gao Su-juan, Xiong Jie 2012, 16 (19):  3560-3567.  doi: 10.3969/j.issn.1673-8225.2012.19.029 Abstract ( 333 )   PDF (366KB) ( 403 )   Save BACKGROUND: Currently, there is lack of effective treating method for stroke. Stem cell transplantation provides a new approach for the treatment of stroke. OBJECTIVE: To multivariately analyze the literature on stem cells transplantation for the treatment of stroke through Scopus database. DESIGN: Bibliometric analysis. DATA RETRIEVAL: A retrieval was performed for the literature of stem cells transplantation for treating stroke during 2006-01 and 2011-12 in Scopus. The retrieval results were analyzed, and the trends were described. SELECTIVE CRITERIA: Articles on stem cells transplantation for the treatment of stroke include the following types: (1)Peer-reviewed original paper; (2) reviews; (3) meeting notes and abstracts; (4) proceeding papers; (5) editorial materials; (6) letters. Exclusive criteria: (1)Articles unrelated the study of stem cells transplantation for the treatment of stroke. (2)Articles published before 2006. MAIN OUTCOME MEASUREMENTS: Country distribution, author distribution, institutional information, journal distribution, as well as citation frequency. RESULTS: A total of 1 045 literatures on stem cells transplantation for the treatment of stroke were retrieved in Scopus database, in which most of paper were published as original articles (n=517). Fifteen articles were identified as classic literatures. The overall number of literature had an upward trend from 2006 to 2011. Currently, United States published most literatures, accounting for 39.8% of all the literatures. Followed by China, which published 88 literatures and accounting for 8.4% of all the published literatures.  CONCLUSION: This paper provides a valuable reference for researchers to understand the overview and present situation of this field.  Related Articles | Metrics

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Stem cells in the treatment of spinal cord injury  Chen Xiao-chun1, Zhang Yang1, Li Zhi1, Zhou Li-nan2 2012, 16 (19):  3568-3575.  doi: 10.3969/j.issn.1673-8225.2012.19.030 Abstract ( 351 )   PDF (360KB) ( 292 )   Save BACKGROUND: Spinal cord injury is a serious trauma to central nervous system. Tissue engineered-cell transplantation provides a new approach for spinal cord injury. OBJECTIVE: To multivariately analyze the literature on neural stem cells transplantation for the treatment of spinal cord injury through Scopus database. DESIGN: Bibliometric analysis. DATA RETRIEVAL: A retrieval was performed for the literature of stem cells transplantation for treating spinal cord injury during 2006-01 and 2011-12 in Scopus. The retrieval results were analyzed, and the trends were described in words and graphics. SELECTIVE CRITERIA: Articles on stem cells transplantation for the treatment of spinal cord injury include the following types:  (1) peer-reviewed original paper; (2) reviews; (3) meeting notes and abstracts; (4) proceeding papers; (5) editorial materials; (6) letters. Exclusive criteria: (1) Articles unrelated the study of stem cells transplantation for the treatment of spinal cord injury. (2) Articles published before 2006. (3) Articles were not published on journals. (4) Articles which need to be retrieved by phone. MAIN OUTCOME MEASUREMENTS: Type of document, authors, publication year, journal distribution, discipline distribution, institutional information, citation frequency and country distribution. RESULTS: A total of 1 080 literatures on stem cells transplantation for the treatment of spinal cord injury were retrieved in Scopus database, in which most of papers were published as original articles (n=607). Fifteen articles were identified as classic literatures. The overall number of literature had an upward trend from 2006 to 2011, and the articles with high citations were published in Journal of Neuroscience. Currently, United States published most literatures, accounting for 28.5% of all the literatures. The second is China, which published 212 literatures, accounting for 19.6% of all the published literatures.  CONCLUSION: This paper provides a valuable reference for researchers to understand the overview and present situation of this field.  Related Articles | Metrics

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Stem cell transplantation for treating ischemic stroke   Chen Juan 2012, 16 (19):  3576-3583.  doi: 10.3969/j.issn.1673-8225.2012.19.031 Abstract ( 292 )   PDF (410KB) ( 602 )   Save BACKGROUND: Ischemic cerebrovascular disease is a serious hazard to human health, which can not be treated effectively at present. How to promote recovery of impaired neural functions for patients with ischemic cerebrovascular disease is the study focus.   OBJECTIVE: To explore the literature data trends in the study of stem cell transplantation for treating ischemic stroke by using search and the depth of analysis capabilities of CNKI database. DESIGN: Bibliometric data analysis DATA RETRIEVAL: A search of related literature of stem cell transplantation for treating ischemic stroke was performed in CNKI database using the key words of “stem cells” “transplantation” and “cerebral ischemia” during 2006-01 to 2011-12. The retrieval results were analyzed, and the trends were described in words and graphics.  SELECTION CRITERIA: Inclusive criteria: Basic research and clinical application of stem cell transplantation for treating ischemic stroke. Exclusive criteria: (1) Literature has nothing to do with the purpose of this review. (2) Duplicated research literature. (3) Journal’s own information. (4) Unpublished papers. (5) The article needs to be retrieved by phone or manual searches. (6) Year book. MAIN OUTCOME MEASUREMENTS: The literatures were analyzed by published year, literature number, subject category, research institutions, source journals, literature citations, literature download frequency, associated literature, distribution of the author, distribution of the funding and major keywords. Additionally, the stem cells from different sources were compared in treating ischemic stroke. RESULTS: A total of 231 research literatures related to stem cell transplantation for treating ischemic stroke were retrieved in CNKI during 2006 to 2011. The number of papers was fluctuated in the past 6 years. Most literatures were published in 2007 and 2009. Bone marrow mesenchymal stem cells, neural stem cells, and amniotic mesenchymal stem cells are the major cells in the study. Journal of Clinical Rehabilitative Tissue Engineering Research published 36 literatures, accounting for 15.6% of all literatures. A total of 44 articles were supported by the National Natural Science Foundation of China. The frequently used key words in stem cell transplantation for treating ischemic stroke are “cerebral ischemia”, “neural stem cells” and “bone marrow mesenchymal stem cells”. CONCLUSION: The bibliometric data analysis on stem cell transplantation for treating ischemic stroke can provide valuable reference for Chinese medical physicians in ischemic stroke field. Related Articles | Metrics

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Effect and mechanism of umbilical cord blood stem cells transplantation in the treatment of stroke  Zhao Hong, Wang Su-ping 2012, 16 (19):  3584-3587.  doi: 10.3969/j.issn.1673-8225.2012.19.032 Abstract ( 380 )   PDF (327KB) ( 385 )   Save BACKGROUND: Many basic and clinical studies have shown that umbilical cord blood stem cells (UCBSCs) transplantation can improve neurological function after focal cerebral ischemia. OBJECTIVE: To summarize the related research and the underlying mechanism of UCBSCs transplantation in the treatment of stroke. METHODS: The first author retrieved PubMed database and Wanfang database for articles of basic research on UCBSCs transplantation for cerebral ischemia published from January 2001 to August 2011. The key words were “umbilical cord blood stem cells, focal cerebral ischemic, transplantation” in English and Chinese. A total of 50 relevant literatures were retrieved, and finally, 30 articles were included according to the inclusion criteria. RESULTS AND CONCLUSION: UCBSCs could be differentiated into nerve cells in appropriate induced conditions with the characters of self-renewal, proliferation, and multi-directional differentiation potential. Related studies had indicated that UCBSCs transplantation could improve neurological disorder and decrease damaged infarct volume. The mechanisms include reducing inflammation, increasing trophic factor secretion, and promoting angiogenesis and nerve fiber reorganization. Greater achievements have been made in the treatment of stroke, but many problems still existed. Some problems need further investigation, for example, how to improve the utilization rate of UCBSCs, prevent excessive cell proliferation and whether these cells can be integrated into the injury site to preserve normal function. Related Articles | Metrics

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Research progress in spinal cord neural stem cell Niche Liao Fa-xue, Zhang Hui, Yin Zong-sheng 2012, 16 (19):  3588-3592.  doi: 10.3969/j.issn.1673-8225.2012.19.033 Abstract ( 283 )   PDF (401KB) ( 343 )   Save BACKGROUND: Spinal cord injury is a common orthopedic disease. A lot of work has been done to focus on neural stem cells for treating spinal cord injury. The effect of spinal cord neural stem cell Niche on neural stem cells has become a gradually increasing research area. OBJECTIVE: To summarize the related research progress in spinal cord neural stem cell Niche. METHODS: A computer-based online search of Pubmed (1980-01/2011-11) and CNKI (2003-01/2011-11) was performed for articles in English and Chinese respectively with the key words “neural stem cells, niche/microenvironment, spinal cord”. In addition, manual search was performed. RESULTS AND CONCLUSION: A total of 483 articles were obtained, and finally 31 articles were included. The Niche provides a relatively stable microenvironment for survival, self-renewal and differentiation of spinal cord neural stem cells. After spinal cord injury, the change of Niche plays an important impact on the fate of spinal cord neural stem cells. Understanding the research progress of spinal cord neural stem cell Niche would provide a theoretical basis for treatment of spinal cord injury using neural stem cells. Related Articles | Metrics

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Maintenance and differentiation of neural stem cells   Gao Hai-xia, Jin Hua-jun, Qian Qi-jun 2012, 16 (19):  3593-3597.  doi: 10.3969/j.issn.1673-8225.2012.19.034 Abstract ( 367 )   PDF (374KB) ( 812 )   Save BACKGROUND: Neural stem cells (NSCs) have great self-renewal capacity and multiple differentiation potential, and therefore have become used to treat neurodegenerative disorders. OBJECTIVE: To summarize the biological characteristics of NSCs and the research progress of NSCs maintenance and differentiation mechanism. METHODS: The PubMed database (http://www.ncbi.nlm.nih.gov/PubMed) (between January 1994 and May 2011) and CNKI database (http://www.cnki.net/index.htm) (between January 2000 and May 2011) were searched by the first author for the articles related to the biological characteristics of NSCs and the maintenance and differentiation mechanism. The key words were “neural stem cells, cell factors, ips, microenvironment” in English and Chinese. Outdated and repetitive studies were excluded, and 55 literatures were included for summarization. RESULTS AND CONCLUSION: Neural stem cells are the original cells with self-renewal capacity and multiple differentiation potential retained during development of the nervous system, Owing to their low-immunogenicity and non-oncogenicty, NSCs are supposed to have great potential in treatment of neurodegenerative diseases. Cell-intrinsic factors such as Tlx, Sox2, Hes and Bmi-1 can regulate the maintenance of self-renewal capacity and multiple differentiation potential of NSCs. Extrinsic signal such as the microenvironment, including the matrix cells around NSCs and cytokines also regulates the maintenance process. Related Articles | Metrics

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Differentiation of mesenchymal stem cells into cardiac-like cells Zhu Sha, Sun Jie, Ling Bin 2012, 16 (19):  3598-3601.  doi: 10.3969/j.issn.1673-8225.2012.19.035 Abstract ( 323 )   PDF (436KB) ( 375 )   Save BACKGROUND: At present, many experiments have shown that mesenchymal stem cells (MSCs) can differentiate into cardiac-like cells or cardiocytes in vitro and in vivo. This makes it possible for MSCs in treatment of cardiac diseases. OBJECTIVE: To compare the laboratory methods and existing problems of differentiation of MSCs into cardiac-like cells. METHODS: A computer-based online search of PubMed database and Wanfang database from December 2000 to December 2010 was performed for relative articles about differentiation of MSCs into cardiac-like cells, with key words of “mesenchymal stem cells, cardiomyocyte, differentiation” in English and in Chinese. RESULTS AND CONCLUSION: The main methods for differentiation of MSCs into cardiac-like cells: (1) conditioned medium: MSCs can be induced to differentiate into cardiocytes in vitro using drugs or in simulated cardiac microenvironment; (2) co-culture with the objective cells: MSCs can be induced to differentiate into cardiocytes using drugs or in cardiac microenvironment without induction. MSCs can differentiate into cardiocytes under cardiac pathological condition, and it makes possible for MSCs in treatment of cardiac diseases, however, more studies are required to improve this. Related Articles | Metrics

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Tissue engineered chondrocytes and bone marrow mesenchymal stem cells for the repair of articular cartilage defects Sun Hao, Zuo Jian 2012, 16 (19):  3602-3605.  doi: 10.3969/j.issn.1673-8225.2012.19.036 Abstract ( 265 )   PDF (423KB) ( 372 )   Save BACKGROUND: As the articular cartilage almost has no self-repair capacity, and in clinic, the repair on it mainly depends on the autologous or allogenic cartilage transplantation, perichondrium or periosteal transplantation and the chondrocytes transplantation. The limitation of autologous cartilage source and the chronic immune rejection of allograft cartilage may eventually lead to the poor prognosis. The cartilage repaired by perichondrium or periosteum transplantation is easy to degenerate which may lead to a poor repair result. OBJECTIVE: To review the research progress of tissue engineered chondrocytes , bone marrow mesenchymal stem cells and the co-culture of them on the repair of allogeneic cartilage defects. METHODS: A computer-based search on the PubMed database and CNKI database from January 1994 to January 2012 was performed for the articles on tissue engineered chondrocytes and bone marrow mesenchymal stem cells for the repair of allograft articular cartilage defects. The English key words were “cartilage defect, allograft, chondrocyte, mesenchymal stem cells，bone marrow mesenchymal stem cells” and the Chinese key words were “cartilage defect, allograft, chondrocyte, bone marrow mesenchymal stem cells”. The repetitive articles and the articles not in English or Chinese were eliminated, and finally, a total of 35 articles were included to review. RESULTS AND CONCLUSION: With the continuous improvement of in vitro cell culture methods, chondrocytes can be isolated from the tough cartilage, and a large number of high-purity chondrocytes and new chondrocytes can be obtained. Due to the low proliferative capacity of the chondrocytes, subculture may easily lead to aging and dedifferentiation; however, the content of bone marrow mesenchymal stem cells is low in adult bone marrow, with the increasing of the passages number, the chondrogenic potential is significantly decreased. When the bone marrow mesenchymal stem cells co-cultured with chondrocytes, they can promote the proliferation and differentiation of each other. And as the seed cells, bone marrow mesenchymal stem cells can reduce the proliferation and passage number of chondrocytes and save the content of chondrocytes, and it can effectively repair the articular cartilage defects when composited with the tissue engineering scaffold.     Related Articles | Metrics

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Characterization of bone marrow mesenchymal stem cells and research progress in treatment of osteoarthritis patients  Zhang Fan, Wang You 2012, 16 (19):  3606-3610.  doi: 10.3969/j.issn.1673-8225.2012.19.037 Abstract ( 355 )   PDF (340KB) ( 379 )   Save BACKGROUND: The basic characteristics and isolation method of bone marrow mesenchymal stem cells as well as the comparison with mesenchymal stem cells from other sources have been well studied. OBJECTIVE: To summarize the basic characteristics of bone marrow mesenchymal stem cells and the research progress in treatment of osteoarthritis patients. METHODS: A computer-based online retrieval of PubMed database was performed by the first author with the key words “MSC, knee” in English. The basic characteristics of mesenchymal stem cells and research progress in clinical treatment have been summarized. In addition, the technique of bone marrow mesenchymal stem cells used for treatment of cartilage defects and treatment progress were also summarized. A total of 93 papers were retrieved. According to inclusion and exclusion criteria, 44 papers were included in the final analysis. RESULTS AND CONCLUSIONS: Mesenchymal stem cells from old people exhibit decreased differentiation potential. Synovia- and bone marrow-derived mesenchymal stem cells exhibit far better chondrogenic ability than muscle- and adipose-derived mesenchymal stem cells. The techniques including bone marrow mesenchymal stem cells transplantation with or without three-dimensional scaffold, intra-articular injection of mesenchymal stem cells, and autologous bone transplantation have been clinically used to treat knee joint cartilage defect and some curative effects have been acquired, but the precise therapeutic methods need further investigation. Related Articles | Metrics