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03 September 2015, Volume 19 Issue 36 Previous Issue    Next Issue

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SPIO and DAPI double labeled bone marrow mesenchymal stem cells of macaques: effects on cell viability and proliferation  Song Qiao-qiao, Zhou Hui-liang, Pan Xing-hua 2015, 19 (36):  5741-5745.  doi: 10.3969/j.issn.2095-4344.2015.36.001 Abstract ( 292 )   PDF (1953KB) ( 560 )   Save BACKGROUND: Traditional cell transplantation tracer methods require histological analysis and identification in vitro, which limits the clinical application of stem cell transplantation. So it is urgent to establish an in vivo noninvasive and repeatable tracer method. OBJECTIVE: To observe the effect of SPIO and DAPI double labeling on survival and proliferation of bone marrow mesenchymal stem cells from macaques. METHODS: Bone marrow mesenchymal stem cells were derived from bone marrow aspirates of healthy macaques using whole bone marrow adherence method. Then, the cells were identified using flow cytometry detection. Bone marrow mesenchymal stem cells were labeled using SPIO and DAPI. Fluorescent microscope was used to detect DAPI positive rate, and Prussian blue staining and transmission electron microscope were employed to measure SPIO positive rate. MTT assay was used to detect cell viability and proliferation. RESULTS AND CONCLUSION: Bone marrow mesenchymal stem cells were successfully isolated from healthy macaques using the whole bone marrow adherence method, and the cell purity was up to 95.1%. SPIO and DAPI  were both successful to label the bone marrow mesenchymal stem cells with a positive rate of 95%-98%, but had no influence on cell viability and proliferation.  中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 Figures and Tables | References | Related Articles | Metrics

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Expressions of apoptosis-related proteins in hepatocyte growth factor-transfected bone marrow mesenchymal stem cells under hypoxia conditions  Cai Wen-qin, Wang Jun-sheng, Su Jin-zi, Jiang Jin-feng, Yao Yue-xian 2015, 19 (36):  5746-5752.  doi: 10.3969/j.issn.2095-4344.2015.36.002 Abstract ( 291 )   PDF (1170KB) ( 360 )   Save BACKGROUND: Previous studies have demonstrated that hepatocyte growth factor (HGF) gene transfection can improve the effectiveness of bone marrow mesenchymal stem cell transplantation, but the mechanism is still unclear. OBJECTIVE: To observe the effects of HGF gene transfection on c-MET, Bax, Bcl-2, Caspase-3 of bone marrow mesenchymal stem cells cultured under hypoxia and serum-free conditions. METHODS: (1) Bone marrow mesenchymal stem cells were isolated and amplified in vitro by differential adhesion method. The infection efficiency of recombinant adenovirus Ad-HGF in bone marrow mesechymal stem  cells was tested by x-gal staining. (2) Bone marrow mesenchymal stem cells were cultured under hypoxia and serum-free conditions for 0, 3, 6, 9, 12 hours. RT-PCR and western blot assays were used to evaluate the expression of Bax, Bcl-2, Caspase-3. (3) Bone marrow mesenchymal stem cells were cultured under hypoxia and serum-free conditions for 6 hours, and RT-PCR and western blot assays were adopted to detect HGF, c-Met, Bax, Bcl-2 and Caspase-3. (4) Cell scratch test was used to detect the effect of HGF transfection on the migration of bone marrow mesenchymal stem cells cultured under hypoxia and serum-free conditions for 6 hours. RESULTS AND CONCLUSION: (1) Transfection efficiency of bone marrow mesenchymal stem cells was increased with multiplicity of infection in a dose-dependent manner. When the multiplicity of infection was 150, the transfection efficiency was 96.4%. (2) Expressions of Bax and Bcl-2 were gradually increased with hypoxia time (P < 0.05). The Bax/Bcl-2 ratio and Caspase-3 expression reach the minimum at 6 hours of hypoxia (P < 0.05). (3) Compared with the control and Ad-LacZ groups, the expressions of HGF, c-Met, Bcl-2 increased, and the expressions of Bax and Caspase-3 decreased in the Ad-HGF group after 6 hours of culture under hypoxia and serum-free conditions (P < 0.05). There was no significant difference between the control and Ad-LacZ groups. (4) The mobility of bone marrow mesenchymal stem cells was higher in the Ad-HGF group than the control group and Ad-LacZ groups after 6 hours of culture under hypoxia (P < 0.05). These findings indicate that transfection of HGF in bone marrow mesenchymal stem cells can increase the expression of c-Met, Bcl-2 and decrease the expression of Bax, Caspase-3 under hypoxia and serum-free conditions, which also enhance the mobility of bone marrow mesenchymal stem cells under hypoxia and serum-free conditions.  中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 Figures and Tables | References | Related Articles | Metrics

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Effect of bone xenograft extract on biological characteristics of bone marrow mesenchymal stem cells Li Fei, Chen Yang, Wang Zhan-chang 2015, 19 (36):  5753-5757.  doi: 10.3969/j.issn.2095-4344.2015.36.003 Abstract ( 301 )   PDF (5650KB) ( 265 )   Save BACKGROUND: How to preserve the biological properties and osteogenic activity of bone xenograft materials after antigen removal has become one of the hot spots. OBJECTIVE: To investigate the effect of bone xenograft extract on proliferation, migration and osteogenic differentiation of bone marrow mesenchymal stem cells. METHODS: Bone marrow mesenchymal stem cells from Sprague-Dawely rats were cultured respectively in bone xenograft extract (experimental group) and DMEM/F12 medium containing 10% fetal bovine serum (control group). MTT, Transwell chamber and flow cytometry were used to detect cell proliferative ability, migration ability and cell cycle changes. Passage 3 bone marrow mesenchymal stem cells were selected in the two groups and cultured in osteogenic induction medium. At 7 and 14 days of osteogenic induction, alkaline phosphatase staining was done; at 21 days of osteogenic induction, alizarin red staining was performed to observe calcium nodule formation. RESULTS AND CONCLUSION: After 1, 3, 5, 7 days of culture, the cell proliferative activity and cell cycle had no significant difference between the two groups, but the number of migrating cells was significantly lower in the control group than the experimental group (P < 0.05). At 7 and 14 days of culture, the alkaline phosphatase level was significantly higher in the experimental group than the control group (P < 0.05); at 21 days of culture, calcium nodes formed in both two groups, but the calcium nodule area was larger in the experimental group than the  control group. These findings indicate that bone xenograft materials have no cytotoxicity but possess good cytocompatibility, which can promote the proliferation, migration and osteogenic differentiation of bone marrow mesenchymal stem cells.  中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 Figures and Tables | References | Related Articles | Metrics

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Expression of aromatase and estrogen-related receptors in human bone marrow mesenchymal stem cells Wei Qiu-shi, Chen Zhen-qiu, He Wei, Deng Wei-min, Wang Hai-bin, Huang Shi-jin, Guo Cheng 2015, 19 (36):  5758-5763.  doi: 10.3969/j.issn.2095-4344.2015.36.004 Abstract ( 263 )   PDF (4783KB) ( 637 )   Save BACKGROUND: Estrogen signaling pathway for interaction between aromatase and estrogen-related receptor may exist in bone marrow mesenchymal stem cells, which is used for regulating biological activity of bone marrow mesenchymal stem cells. OBJECTIVE: To observe the expression of aromatase and estrogen-related receptors in adult bone marrow mesenchymal stem cells during osteogenic differentiation. METHODS: Bone marrow mesenchymal stem cells were respectively cultured in low-glucose DMEM medium (control group) and osteogenic induction medium (induction group). Cell proliferation and calcium deposition were determined by MTT assay and alizarin red staining, respectively. The expression of aromatase, estrogen receptor α, estrogen receptor β, and estrogen-related receptor α during osteogenic differentiation were determined by real-time PCR and western blot analysis. Estradiol levels in supernatants and lysates were detected by ELISA method. RESULTS AND CONCLUSION: In the induction group, the proliferation ability of bone marrow mesenchymal stem cells was the strongest at 72 hours of culture; while there were a great amount of calcium nodules formed at 21 days of culture. Results from PCR and western blot assay showed that the expression of aromatase and estrogen receptor α was improved in the induction group, but the expression of estrogen-related receptor α was inhibited. There was no difference in the expression of estrogen receptor β between the two groups. ELISA results indicated that the level of estradiol in the supernatant of induction group was the highest. These findings indicate that aromatase, estrogen receptor α, estrogen receptor β and estrogen-related receptor α are all involved in osteogenesis of bone marrow mesenchymal stem cells. Moreover, estradiol can be synthesized and secreted in bone marrow mesenchymal stem cells, and most likely, promote the osteogenic differentiation of bone marrow mesenchymal stem cells by related receptor pathway.  中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 Figures and Tables | References | Related Articles | Metrics

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Culture supernatant of bone marrow mesenchymal stem cells for treatment of bronchial asthma and its influence on lung inflammation Lei Yan, Liu Chang-qing 2015, 19 (36):  5764-5768.  doi: 10.3969/j.issn.2095-4344.2015.36.005 Abstract ( 398 )   PDF (4742KB) ( 401 )   Save BACKGROUND: Bronchial asthma is considered generally to have an association with Th2 immune response disease, but there is no ideal treatment. Bone marrow mesenchymal stem cells as a kind of adult stem cells not only have the multipotent differentiation and proliferation capacity, but also have low immunogenicity and immunoregulation ability. OBJECTIVE: To investigate the effect of culture supernatant of bone marrow mesenchymal stem cells on lung inflammation of bronchial asthma mice. METHODS: Twenty experimental mice were randomly divided into control and experimental groups, 10 mice in each group. At 0 and 14 days, intraperitoneal injection of ovalbumin induced sensitization in mice, and at 24-26 days, aerosolized ovalbumin solution was used for excitation. From the 24th day, in the experimental group, bone marrow mesenchymal stem cell supernatant was intraperitoneally injected at 2 hours before excitation; meanwhile, normal saline was injected in the control group. At the last of excitation, the mice were sacrificed under anesthesia to take serum samples, bronchoalveolar lavage fluid and lung tissues. RESULTS AND CONCLUSION: (1) Mice in the control group appeared to have abnormal lung tissue structure, and there were a large amount of eosinophils and monocytes in the submucosa and muscularis. However, lung inflammation was relieved in the experimental group after bone marrow mesenchymal stem cell treatment.  (2) The levels of interleukin-17 in the bronchoalveolar lavage fluid and serum were significantly lower in the experimental group than the control group (P < 0.05), but there was no difference in the levels of interleukin-4 between the two groups (P > 0.05). These findings indicate that the intraperitoneal injection of bone marrow mesenchymal stem cells can ease lung inflammation and reduce levels of inflammatory markers in the bronchoalveolar lavage fluid and serum of bronchial asthma mice.  中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 Figures and Tables | References | Related Articles | Metrics

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Biocompatibility of bone marrow mesenchymal stem cells with bladder acellular matrix scaffold Zhao Xiao-jun, Yu Jun, Duan Ying-fei 2015, 19 (36):  5769-5773.  doi: 10.3969/j.issn.2095-4344.2015.36.006 Abstract ( 306 )   PDF (931KB) ( 387 )   Save BACKGROUND: In the repair of urinary tract defects, we have been actively trying to construct the urinary tract substitutes with normal physiological function through combining ideal seed cells and proper scaffold materials by tissue engineering method. OBJECTIVE: To investigate the biocompatibility of bone marrow mesenchymal stem cells with rabbit bladder acellular matrix scaffold. METHODS: Rabbit bone marrow mesenchymal stem cells were isolated and cultured using density gradient centrifugation method. Passage 3 rabbit bone marrow mesenchymal stem cells were cultured on the rabbit bladder acellular matrix. The cells were counted every day for 12 days, to drawn a cell growth curve. Bone marrow mesenchymal stem cells cultured alone were used as control group. RESULTS AND CONCLUSION: Bone marrow mesenchymal stem cells were successfully seeded onto the bladder acellular matrix. Under the inverted microscope, the cells grew out of the bladder acellular matrix, and a great amount of long spindle-shaped cells were found around the bladder acellular matrix. With 5 days of inoculation, the cells in the two groups grew gently; at 6-9 days, the cell growth curve gradually became steeper, and the cell division and growth were increased exponentially; at 10-12 days, the cells recovered to a gentle state. Cell growth curves in the two groups were basically coincident, suggesting that rabbit bone marrow mesenchymalstem cells have good biocompatibility with the bladder matrix.  中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 References | Related Articles | Metrics

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Interventional effects of different Chinese medicine therapies on proliferation and differentiation of bone marrow mesenchymal stem cells homing to the border zone of myocardial infarction  Zhang Jin-sheng, Zhang Bao-xia, Zhang Yang-yang, Zhu Hui-fang 2015, 19 (36):  5774-5781.  doi: 10.3969/j.issn.2095-4344.2015.36.007 Abstract ( 271 )   PDF (2733KB) ( 306 )   Save BACKGROUND: New ideas for regenerative treatment of cardiovascular disease are as follows: to understand the changes in all kinds of stem cells differentiating into cardiomyocytes, to determine the physiological role of differentiated stem cells in cardiac functional activities, to stimulate the proliferative potential of various stem cells under certain conditions that can directly differentiate into functional cardiomyocytes, thereby replenishing deficient cardiomyocytes and effectively inhibiting excessive proliferation of fibroblasts. OBJECTIVE: To investigate the effects of panax notoginseng saponins, salidroside and astragalus effective components on the homing, proliferation and differentiation of endogenous CD105-positive bone marrow mesenchymal stem cells of myocardial infarction rats. METHODS: Acute myocardial infarction rats were randomly divided into activating blood circulation group, tonifying qi group, activating blood circulation+tonifying qi group (combined group), model group. Normal rats served as control group. The former three groups were orally given 80 g/L Xuesaitong soft capsules (panax notoginseng saponins as the main component), 0.5 g/mL astragalus particles and 70 g/L Nuodikang capsules (salidroside as the main component), respectively, at a dose of 1 mL/100 g, once a day, totally for 28 days. The control and model groups were given the same volume of normal saline. Expressions of CD105, CD117, vimentin, cardiac troponin T (cTnT), zinc finger transcription factor 4 (GATA-4) and Ki-67 were detected using immunohistochemical method at 1, 3, 7, 14, 28 days of drug administration. RESULTS AND CONCLUSION: (1) Compared with the control group, the expressions of CD105, CD117, cTnT, GATA-4 and Ki-67 were higher in the model group, which were increased at 1-7 days, peaked at 7 days, and then decreased. Compared with the model group, the expressions of CD105, CD117, cTnT, GATA-4 and Ki-67 were significantly higher in the activating blood circulation group, tonifying qi group and combined group, especially in the activating blood circulating group, at different time of drug administration (P < 0.05). In each group, the staining results of Ki-67 were not exactly parallel to those of CD105, CD117, cTnT and GATA-4, but their rising tendency was substantially the same. (2) Compared with the control group, the vimentin expression in the model group was higher, which showed an increasing tend at 1-3 days, peaked at 3 days and then declined. Compared with the model group, the vimentin expression was significantly lower in the activating blood circulation group, tonifying qi group and combined group, especially in the activating blood circulating group, at different time of drug administration (P < 0.05). It suggested that the activating blood circulation group had a remarkable antifibrotic role. These findings indicate that panax notoginseng saponins, salidroside and astragalus effective components all can promote the proliferation and differentiation of bone marrow mesenchymal stem cells to delay myocardial remodeling, but the recipe for promoting blood circulation has the most obvious outcomes.   中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 Figures and Tables | References | Related Articles | Metrics

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Vascular endothelial growth factor 165 transfection promotes proliferation of adipose-derived mesenchymal stem cells  San Guang, Song Jia 2015, 19 (36):  5782-5788.  doi: 10.3969/j.issn.2095-4344.2015.36.008 Abstract ( 260 )   PDF (6887KB) ( 670 )   Save BACKGROUND: Vascular endothelial growth factor is the most important pro-angiogenic factor, which can promote revascularization and survival of fat grafts during fat transplantation. OBJECTIVE: To investigate the effect of vascular endothelial growth factor 165 transfection on the proliferation of adipose-derived mesenchymal stem cells. METHODS: Recombinant vascular endothelial growth factor 165 mRNA fragment was transmitted into adenovirus pAdEasy-1 systems that were packaged to measure viral titer (experimental group). Empty adenovirus was also packaged as control group. Two kinds of packaged adenovirus solution were transferred into adipose-derived mesenchymal stem cells at the best multiplicity of infection=100. Cells with no transfection served as blank group. RT-PCR and western blot methods were used to detect the expression of vascular endothelial growth factor 165 at mRNA and protein levels; MTT method was adopted to detect cell proliferation. RESULTS AND CONCLUSION: The expressions of vascular endothelial growth factor 165 mRNA and protein were significantly higher in the experimental group than the control and blank groups (P < 0.05). The division and proliferation of transfected adipose-derived mesenchymal stem cells were increased significantly in the experimental group, which was significantly different from the control and blank groups (P < 0.05). These finding indicate that vascular endothelial growth factor 165 transfection cannot only sustain the target protein expression of adipose- derived mesenchymal stem cells, but also promote the proliferation of adipose-derived mesenchymal stem cells remarkably.   中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 Figures and Tables | References | Related Articles | Metrics

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Isolation, culture and biological characteristics of adipose-derived mesenchymal stem cells from patients with hepatitis B virus infection  Wang Yan-hui, Zhang Shu-qin, Zhao Wen-jing 2015, 19 (36):  5789-5794.  doi: 10.3969/j.issn.2095-4344.2015.36.009 Abstract ( 222 )   PDF (5466KB) ( 481 )   Save BACKGROUND: Chronic hepatitis B virus infection can impact the biological characteristics of bone marrow mesenchymal stem cells. Adipose-derived mesenchymal stem cells have gained more and more attention due to their high safety, little invasiveness, easy purification and rapid proliferation. OBJECTIVE: To establish the isolation and culture method of adipose-derived mesenchymal stem cells from patients with hepatitis B virus infection in vitro, and to observe the biological characteristics of cells. METHODS: Adipose-derived mesenchymal stem cells were isolated from the subcutaneous fat of hepatitis B virus infection patients by collagenase digestion and adherent method. Growth curve of adipose-derived mesenchymal stem cells were detected by MTT method and cell phenotypes were detected by flow cytometry. The adipogenic and osteogenic differentiation potential of adipose-derived mesenchymal stem cells were detected in vitro. RESULTS AND CONCLUSION: (1) Adipose-derived mesenchymal stem cells from 10 patients with hepatitis B virus infection were all isolated and cultured successfully. The primary passage time of adipose-derived mesenchymal stem cells was (8.3±1.2) days. The growth curve of cells was “S” shaped. Cells came into a logarithmic phase at days 3-4, and came into platform at day 7. (3) Passage 3 adipose-derived mesenchymal stem cells highly expressed CD29, CD166, HLA-ABC and CD44, but did not express or lowly expressed CD34 and HLA-DR. (3) Adipose-derived mesenchymal stem cells differentiated into adipocytes after adipogenic induction, and differentiated cells were positive for oil red O staining; after osteogenic induction, adipose-derived mesenchymal stem cells differentiated into osteoblasts that were positive for alkaline phosphatase staining. These findings indicate that the collagenase digestion and adherent method can be used to effectively isolate adipose-derived mesenchymal stem cells from patients with hepatitis B virus infection, and the cell proliferation is rapid so that a large number of cells can be obtained in the short term. 中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 Figures and Tables | References | Related Articles | Metrics

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Bone marrow mesenchymal stem cell transplantation for cerebral infarction: CT and MRI characteristics Yang Ting-shuang 2015, 19 (36):  5795-5799.  doi: 10.3969/j.issn.2095-4344.2015.36.010 Abstract ( 337 )   PDF (4308KB) ( 348 )   Save BACKGROUND: Studies have shown that bone marrow mesenchymal stem cells can play a catalytic role in brain function recovery through a variety of ways. OBJECTIVE: To analyze the CT and MRI features of rats with cerebral infarction undergoing bone marrow mesenchymal stem cell transplantation. METHODS: Forty rats with cerebral infarction were randomized into two groups: model group was treated with 1 mL PBS and transplantation group was injected with 2.0×109/L cell suspension. Modified neurological severity score was detected at 1, 2, 3 weeks after transplantation for neurological function evaluation. CT and MRI scans were performed at 6 hours, 1, 3, 5, 7 days after transplantation to observe the changing features of T1WI, T2WI, FLAIR, DWI sequences in the infracted area as well as infarction size. Signal intensity ratios (SIR) of T2WI, FLAIR, DWI sequences and relative rates of change (?SIR) were determined and compared with the normal values. RESULTS AND CONCLUSION: After transplantation 1, 2 and 3 weeks, the modified neurological severity scores were significantly lower in the transplantation group than the model group (P < 0.05). After transplantation 3, 5, 7 days, the infracted size was reduced significantly in the transplantation group compared with the model group (P < 0.05). Compared with the model group, SIR value of T1WI sequence was higher, but the SIR values of T2WI  and FLAIR sequences were lower in the transplantation (P < 0.05). There were significant differences in the ?SIR values of T2WI, FLAIR and DWI (P < 0.05) rather than T1WI (P > 0.05) between the two groups. These findings indicate that MRI can show the brain sections at any angle, and play an important role to assess the therapeutic efficacy of bone marrow mesenchymal stem cell transplantation on cerebral infarction. 中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程   Figures and Tables | References | Related Articles | Metrics

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Mechanism by which interferon reduces the resistance of MGMT positive glioma stem cells to temozolomide Su Hui, Liu Zhao-wei, Du Hong-li, Qiao Yan-mei 2015, 19 (36):  5800-5805.  doi: 10.3969/j.issn.2095-4344.2015.36.011 Abstract ( 423 )   PDF (1078KB) ( 363 )   Save BACKGROUND: It is easy to appear the phenomenon of drug resistance in the treatment of patients with glioma, and O(6)-methylguanine-DNA methyltransferase (MGMT) is a kind of multidrug resistance gene expressed in glioma. OBJECTIVE: To investigate the role of interferon to increase the sensibilization of MGMT positive glioma stem cells to temozolomide in vitro. METHODS: Glioma cell lines, U251 and SKMG-4, were induced by suspended cloning ball formation method to harvest MGMT positive glioma stem cells, U251G and SKMG-4G. Cell counting kit-8 assay was used to detect the killing effect of interferon α/β combined with temozolomide on MGMT positive glioma stem cells. RT-PCR and western blot assay were employed to determine the expression of MGMT and nuclear factor κB in MGMT positive glioma stem cells. RESULTS AND CONCLUSION: Western blot results showed positive expression of MGMT in U251G and SKMG-4G cells at protein levels. After intervention with interferon α/β, the mRNA expression of MGMT and nuclear factor κB in SKMG-4G and U251G cells was reduced significantly, and then further decreased after temozolomide treatment. These findings indicate that interferon α/β can remarkably strengthen the killing effect of temozolomide on MGMT positive glioma stem cells. 中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 Figures and Tables | References | Related Articles | Metrics

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Isolation, culture and in vitro proliferation of breast cancer stem cells after different cycles of neoadjuvant chemotherapy  Liu Wei, Wei Dong-dong, Han Li-jie 2015, 19 (36):  5806-5810.  doi: 10.3969/j.issn.2095-4344.2015.36.012 Abstract ( 216 )   PDF (797KB) ( 379 )   Save BACKGROUND: Whether there are breast cancer stem cell microspheres in the breast cancer tissues and whether these microspheres have an impact on isolation and culture of breast cancer stem cells after different cycles of neoadjuvant chemotherapy are still unclear. OBJECTIVE: To explore the proliferation and differentiation of breast cancer stem cells in breast cancer tissues after different cycles of neoadjuvant chemotherapy. METHODS: Breast cancer stem cell microspheres were isolated from the breast cancer tissues after different cycles of neoadjuvant chemotherapy to drawn a cell growth curve. Immunocytochemical method was used to detect ALDH1 expression.  RESULTS AND CONCLUSION: Microspheres could be obtained from the specimens of neoadjuvant chemotherapy for two, three and four cycles rather than one cycle. At 3 days prior to culture, there was no difference in the number of cells isolated after two- and three-cycle neoadjuvant chemotherapy; but after 3 days, the cells from the three-cycle neoadjuvant chemotherapy proliferated faster than those from the two-cycle neoadjuvant chemotherapy; after 6 days, the cell growth curve of two-cycle neoadjuvant chemotherapy was in the plateau stage, and the proliferation of cells from the three-cycle neoadjuvant chemotherapy showed a rapid increase trend. The positive expression of ALDH1 in the microspheres from the three-cycle neoadjuvant chemotherapy was higher than that from the two-cycle neoadjuvant chemotherapy. These findings indicate that breast cancer stem cells from the specimens of two- and three-cycle neoadjuvant chemotherapy both have proliferation and differentiation potentials, and the specimens of three-cycle neoadjuvant chemotherapy or above are preferred.    中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 Figures and Tables | References | Related Articles | Metrics

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Ganglioside combined with bone marrow mesenchymal stem cells transplantation for treatment of traumatic brain injury  Huang Yong-hui 2015, 19 (36):  5811-5815.  doi: 10.3969/j.issn.2095-4344.2015.36.013 Abstract ( 166 )   PDF (889KB) ( 370 )   Save BACKGROUND: Bone marrow mesenchymal stem cells can secrete neurotrophic factors in vitro, and can also be differentiated into neurons, thereby contributing to the repair of traumatic brain injury. However, the short life cycle of bone marrow mesenchymal stem cells influences their protective effects on the damaged brain tissues. OBJECTIVE: To observe the effect of bone marrow mesenchymal stem cell transplantation combined with ganglioside in rats with traumatic brain injury. METHODS: Sixty Wistar rats were used to make severe traumatic brain injury models using a hydraulic head injury instrument, and then randomized into three groups: 1 mL DMEM, 1 mL bone marrow mesenchymal stem cell suspension (1×1010/L), 1 mL bone marrow mesenchymal stem cell suspension (1×1010/L) combined with ganglioside solution (30 mg/kg) were injected respectively in model group, transplantation group and combined group, once a day, totally for 3 days. Neurological behavior scores were observed according to Longa method at 24 hours after modeling and at 3 days, 1, 2, 3, 4 weeks after cell transplantation. At 3 days after cell transplantation, RT-PCR and western blot assay were employed to detect aquaporin 4 mRNA and protein expressions. At 1 week after transplantation, hematoxylin-eosin staining was performed for pathological observation of the damaged brain tissues. RESULTS AND CONCLUSION: At 3 days, 1, 2, 3, 4 weeks after cell transplantation, the neurological behavior scores were ranked as follows: combined group < transplantation group < model group (P < 0.05). At 3 days after cell transplantation, the mRNA and protein expression of aquaporin 4 was ranked as follows: model group > transplantation group > combined group (P < 0.05). Hematoxylin-eosin staining showed that the recovery of brain tissue was better in the combined group than the model and transplantation groups (P < 0.05). These findings indicate that bone marrow mesenchymal stem cells combined with ganglioside can significantly improve the neurological behavior function of rats with traumatic brain injury. 中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 Figures and Tables | References | Related Articles | Metrics

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The expression of CD133 in non-small cell lung cancer  Du Jin, Gao Jian-hui, Han Ji-chang, Li Hong-bing, Cheng Xiang-shu 2015, 19 (36):  5816-5820.  doi: 10.3969/j.issn.2095-4344.2015.36.014 Abstract ( 346 )   PDF (767KB) ( 434 )   Save BACKGROUND: The CD133 expression in non-small cell lung cancer shows some changes, which is definitely related to the occurrence and development of diseases. OBJECTIVE: To investigate the expression of CD133 in non-small cell lung cancer, and to analyze its relationship with clinical pathological factors and prognosis. METHODS: Non-small cell lung cancer tissue specimens from 135 cases were collected, and normal lung tissue specimens from 60 cases were set as normal control group. Immunohistochemical staining was used to detect CD133 expression in two groups, and the relationship between the expression of CD133 protein and the clinical pathological factors was analyzed. RESULTS AND CONCLUSION: (1) The positive expression of CD133 protein in the non-small cell lung cancer group was significantly higher than that in the normal control group (P < 0.05). (2) CD133 protein expression had no association with age, gender, tumor size, tumor location, histological type (P > 0.05), and CD133 protein expression was significantly increased with the differentiation of non-small cell lung cancer (P < 0.05). The positive expression rate of CD133 protein was significantly different between different clinical stages and lymph node metastasis (P < 0.05). (3) CD133 and TNM staging were independent prognostic factors for non-small cell lung cancer (P < 0.05), and the median survival time was significantly shorter in the positive group than in the  negative group (P < 0.05). The results indicate that CD133 is involved in the occurrence, development, infiltration and metastasis of non-small cell lung cancer, and it hasimportant clinical significance for the disease progression and prognosis. 中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 Figures and Tables | References | Related Articles | Metrics

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Haploidentical allogeneic hematopoietic stem cell transplantation for severe aplastic anemia: a retrospective analysis  Tong Chun, Guo Zhi, Lou Jin-xing, Liu Xiao-dong, Yang Kai, He Xue-peng, Zhang Yuan, Chen Peng, Chen Hui-ren 2015, 19 (36):  5821-5826.  doi: 10.3969/j.issn.2095-4344.2015.36.015 Abstract ( 211 )   PDF (945KB) ( 394 )   Save BACKGROUND: Allogeneic hematopoietic stem cell transplantation is currently recognized as the first-line therapy for severe aplastic anemia. However, with the popularity of the one-child families, the source of fully matched hematopoietic stem cell transplantation is limited, so haploidentical hematopoietic stem cell transplantation is favored.  OBJECTIVE: To retrospectively compare and analyze the clinical efficacy and safety of haploidentical allogeneic hematopoietic stem cell transplantation and fully matched hematopoietic stem cell transplantation for the treatment of severe aplastic anemia. METHODS: Clinical data of 15 patients with severe aplastic anemia (treatment group) who underwent haploidentical allogeneic hematopoietic stem cell transplantation in the Department of Hematology General Hospital of Beijing Military Region from January 2013 to January 2015 were retrospectively analyzed. Pretreatment regimen was cyclophosphamide, fludarabine, Busulfex, combined with anti-human lymphocyte immune globulin. Donors received granulocyte colony-stimulating factor, and the transplantation method was bone marrow mobilization combined with peripheral blood stem cell transplantation. Combined immunosuppressive agents including cyclosporine A, methotrexate, tacrolimus, were adopted for prevention of graft versus host disease. Another 15 cases of severe aplastic anemia undergoing fully matched hematopoietic stem cell transplantation served as control group over the same period. Complications and survival of the two groups were statistically analyzed. RESULTS AND CONCLUSION: By the end of July 2015, the median follow-up time of the treatment group was 20.7 months (6-30 months), and hematopoietic reconstruction was achieved in all cases, including four cases of graft versus host disease, five cases of pulmonary infection, three cases of sepsis, and one case died of pulmonary infection, one cases died of sepsis, and two cases died of graft versus host disease. In the control group, the median follow-up time was 19.7 months (5-28 months), hematopoietic reconstruction was achieved in all cases. There were three cases of graft versus host disease, four cases of pulmonary infection, one case died of pulmonary infection, and two cases died of graft versus host disease. The total survival rates of the two groups were 73% and 80% respectively, with no significant difference (P=0.67). The haploidentical allogeneic hematopoietic stem cell transplantation for severe aplastic anemia is safe and effective, and the clinical efficacy is comparable to the fully matched hematopoietic stem cell transplantation. 中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 Figures and Tables | References | Related Articles | Metrics

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Telomerase reverse transcriptase gene transfection of human amniotic electrical mesenchymal stem cell transplantation for treatment of diabetes mellitus  Fu Jian-ru 2015, 19 (36):  5827-5832.  doi: 10.3969/j.issn.2095-4344.2015.36.016 Abstract ( 237 )   PDF (1055KB) ( 373 )   Save BACKGROUND: The amniotic membrane is the rejected material after birth. Amniotic mesenchymal stem cells are characterized as easy harvesting, strong proliferation ability, no ethical controversy, and low immunogenicity. OBJECTIVE: To electrotransfer human telomerase reverse transcriptase (hTERT) gene into amniotic mesenchymal stem cells transplanted into diabetic rats and to explore its effect on diabetic rats. METHODS: Human amniotic mesenchymal stem cells were isolated, cultured and electrotransferred by hTERT gene. Ten of 50 Sprague-Dawley rats were randomized selected as controls, and the remaining rats were used to establish diabetic models through injection of 45 mg/kg streptozotocin. Thirty-six model rats were randomized into model group, cell transplantation group and hTERT-transfected cell transplantation group, with 12 rats in each group. In the latter two groups, human amniotic mesenchymal stem cells and hTERT-transfected amniotic mesenchymal stem cells were injected via sublingual veins, respectively. After transplantation, blood glucose levels were monitored dynamically, and plasma insulin concentration was detected every week. Pancreas tissues were taken and cut into sections for histological observation using hematoxylin-eosin staining. RESULTS AND CONCLUSION: At 4 weeks after transplantation, the blood glucose levels were significantly lower in the two cell transplantation groups than the model group (P < 0.05), and especially in the hTERT-transfected cell  transplantation group, the blood glucose level was close to the normal value (P > 0.05). However, the model group still had a higher blood glucose level. At 6 weeks after transplantation, compared with the model group, the plasma insulin concentration was significantly increased in the two cell transplantation group (P < 0.05), and the severity of pancreatic injury was also eased in these two groups (P < 0.05), especially in the hTERT-transfected cell transplantation group (P < 0.05). These findings indicate that hTERT-transfected amniotic mesenchymal stem cell transplantation can dramatically decrease blood glucose level and relieve pancreatic injury in diabetic rats, which is an effective method for treatment of mellitus diabetes in rats. 中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 Figures and Tables | References | Related Articles | Metrics

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Effect of umbilical cord blood stem cells on blood glucose levels and PDX-1 and MafA levels in type 1 diabetic rats Du Jun-wen, Wu Tao, Zhang Kun, Su Bai-yu, Lu Cai-ping, Wang Wei-chao, Lei Lin, Guo Jing-xia 2015, 19 (36):  5833-5837.  doi: 10.3969/j.issn.2095-4344.2015.36.017 Abstract ( 278 )   PDF (903KB) ( 489 )   Save BACKGROUND: Type 1 diabetes mellitus is an autoimmune disease, which is characterized as the selective destruction of pancreatic beta cells in the body, resulting in the lack of insulin secretion. Umbilical cord blood stem cells have the potential to differentiate into islet cells in vitro and in vivo, which play a certain hypoglycemic effect. OBJECTIVE: To explore the effects of umbilical cord blood stem cells on blood glucose levels and PDX-1 and MafA levels in the pancreatic tissue of type 1 diabetic rats. METHODS: Thirty Sprague-Dawley rats were randomly divided into three groups, with 10 rats in each group. In treatment and model groups, type 1 diabetes mellitus modes were established. After modeling, the treatment group was given a single tail vein injection of umbilical cord blood stem cells; the normal control group was given the same volume of normal saline; the model group was given the same volume of umbilical cord blood stem cell buffer solution. Oral glucose tolerance test was adopted to assess the islet function of rats; hematoxylin-eosin staining was used to observe the pancreatic morphology of rats; western blot and PCR methods were employed to detect expressions of PDX-1 and MafA in pancreatic tissues at protein and mRNA levels. RESULTS AND CONCLUSION: (1) Compared with the normal control group, the levels of blood glucose in the model and treatment groups were significantly higher at 0, 30, 60, 90 minutes (P < 0.05). At 120 minutes, the blood glucose level in the model group was significantly higher than that in the normal control group (P < 0.05), but there was no difference between the treatment and normal control groups (P > 0.05). (2) The number of islets in the model group was decreased, and the boundary was unclear and irregular; in the treatment group, the number of islets was decreased, but the boundary was still clear.(3) The expressions of PDX-1 and MafA in the treatment group were similar to those in the normal control group (P > 0.05), but significantly higher than those in the model group (P < 0.05). These findings suggest that the umbilical cord blood stem cell transplantation can significantly reduce the blood glucose levels in type 1 diabetic rats, improve the function of islet and morphology of pancreas, and up-reuglate the expressions of PDX-1 and MafA. 中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 Figures and Tables | References | Related Articles | Metrics

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Effects of midbrain neural stem cells and bone marrow stromal stem cells on behaviors and brain morphology of rats with Parkinson’s disease  Liang Xin-ming, Fu Guo-hui, Zhang Bao-chao 2015, 19 (36):  5838-5842.  doi: 10.3969/j.issn.2095-4344.2015.36.018 Abstract ( 260 )   PDF (4256KB) ( 603 )   Save BACKGROUND: For treatment of central nervous system diseases, neural stem cells (NSCs) or bone marrow stromal stem cells (BMSCs) can be transplanted into the brain, but there are less reports to compare the effects of two kinds of stem cell transplantation. OBJECTIVE: To explore the effect of midbrain NSCs and BMSCs on the behavior and brain morphology of rats with Parkinson’s disease. METHODS: Fifty-eight Sprague-Dawley rats were enrolled to establish Parkinson’s disease models, and then randomly divided into three groups, which were treated with 5 μL midbrain NSCs (n=20), 5 μL BMSCs (n=20) and 5 μL normal saline (n=18) via two coordinate points of the right striatum at 3 weeks after modeling, respectively. At 5 months after transplantation, the rats underwent intraperitoneal injection of apomorphine to observe behavioral changes, and then, the striatum was taken for immunohistochemistry staining. RESULTS AND CONCLUSION: The number of rotations was reduced significantly in the BMSCs and midbrain NSCs groups at 5 months after transplantation (P < 0.05), which was significantly lower than that in the normal saline group (P < 0.05). However, there was no significant difference between the BMSCs and NSCs groups (P > 0.05). In the BMSCs group, BrdU/Nestin positive cells were seen in the brain stratium at 1 week after transplantation; BrdU/GFAP and BrdU/NSE positive cells as well as TH positive cells rather than BrdU/TH positive cells were found in the brain stratium at 1 month after transplantation; after that, the number of BrdU/Nestin positive cells was reduced gradually and disappeared ultimately, but there were still a certain number of BrdU/GFAP and BrdU/NSE positive cells, especially the former ones. Meanwhile, the NSCs group also had a similar situation, but no double-labeled cells were in the normal saline group. These findings indicate that midbrain NSCs and BMSCs transplantation can both improve the behavior of Parkinson’s disease rats, and differentiate into neurons, astrocytes and dopaminergic neurons. 中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 Figures and Tables | References | Related Articles | Metrics

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Liu Zhen-hua, Studying for doctorate, Chief physician, Department of Neurology, Shandong Provincial Liu Zhen-hua, Wang Shi-jun 2015, 19 (36):  5843-5847.  doi: 10.3969/j.issn.2095-4344.2015.36.019 Abstract ( 274 )   PDF (4576KB) ( 600 )   Save BACKGROUND: In recent years, cell transplantation therapy is widely used in the treatment of Parkinson’s disease with satisfactory outcomes. OBJECTIVE: To investigate the proliferation and differentiation of transplanted neural stem cells in Parkinson’s disease rats, thereby providing a new theoretical basis for the treatment of Parkinson’s disease. METHODS: Neural stem cells from the brain tissues of rats were isolated and cultured in vitro using collagenase digestion method. Unilateral rat model of Parkinson’s disease was made by 6-hydroxydopamine method, and the successful rat models were divided into model group and cell transplantation group, with seven rats in each group. The rats in the model group were given 4 mL normal saline at the damaged striatum, and those in the cell transplantation group were injected 5 μL green fluorescent protein-labeled neural stem cells (1×109/L) into the damaged striatum. RESULTS AND CONCLUSION: After cell transplantation, the positive area of tyrosine hydroxylase was significantly lower in the cell transplantation group than the model group. Compared with the model group, the expression level of Ptx3 mRNA was significantly higher in the cell transplantation group at 7, 14, 28 days after cell transplantation and the expression of SHH mRNA was significantly higher in the cell transplantation group at 14 and 28 days after cell transplantation. However, there was no difference in the expression of Nurrl mRNA between the two groups. These findings indicate that neural stem cells injected into the damaged striatum of rats can differentiate into dopaminergic neurons, which bring a breakthrough in the treatment of Parkinson’s disease. 中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 Figures and Tables | References | Related Articles | Metrics

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A static magnetic field with certain intensity can promote the secretion of nerve growth factor from Schwann cells Wei Ling-yun, Wang Rui-ying, Tang Ji-cun 2015, 19 (36):  5848-5852.  doi: 10.3969/j.issn.2095-4344.2015.36.020 Abstract ( 271 )   PDF (5496KB) ( 416 )   Save BACKGROUND: There are less studies on static magnetic field, and its biological effects are still unclear. OBJECTIVE: To investigate the static magnetic field effect on the secretion of nerve growth factor from Schwann cells.  METHODS: Schwann cells were randomly assigned into three groups: 0.05 mT sciatic magnetic field group, 0.1 mT sciatic magnetic field group and control group. At 2 days of inoculation, the cells were exposed to the sciatic magnetic field, 12 hours per day. Cells in the control group were not exposed to the sciatic magnetic field. After 6 days of exposure, RT-PCR method was used to detect nerve growth factor mRNA expression in Schwann cells, and ELISA was adopted to detect the level of nerve growth factor secreted from Schwann cells. RESULTS AND CONCLUSION: The mRNA expression and secreted level of nerve growth factors in the culture supernatant were significantly higher in the two sciatic magnetic field groups than the control group (P < 0.05), especially in the 0.1 mT group. However, there was no difference between the 0.05 mT and 0.1 mT groups (P > 0.05). These findings indicate that the static magnetic field with certain intensity can promote the secretion of nerve growth factors from Schwann cells. 中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 Figures and Tables | References | Related Articles | Metrics

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Transfection of adeno-associated virus encoding beta-nerve growth factor into endothelial progenitor cells  Yang Zhong-yan, Gao Chun-zheng, Wu Dong-jin, Peng Chang-liang 2015, 19 (36):  5853-5858.  doi: 10.3969/j.issn.2095-4344.2015.36.021 Abstract ( 299 )   PDF (5979KB) ( 335 )   Save BACKGROUND: Nowadays, gene therapy has become a new trend for disease therapy and brought promise for some refractory diseases. Its key is to choose proper cells, genes and vectors. OBJECTIVE: To use recombinant adeno-associated virus mediated β-nerve growth factor (β-NGF) to transfect rat bone marrow-derived endothelial progenitor cells in vitro, and to investigate the effect of β-NGF expression on the proliferation of endothelial progenitor cells. METHODS: The endothelial progenitor cells were isolated, cultured and identified from the bone marrow of rats. Empty vector or recombinant adenovirus-associated virus containing β-NGF gene was transferred into endothelial progenitor cells. We examined the transfection efficiency by fluorescence expression of green fluorescent protein. Expression of β-NGF protein was detected using ELISA, and its effect on the proliferation of endothelial progenitor cells was determined using MTT method. RESULTS AND CONCLUSION: Rat endothelial progenitor cells were isolated and cultured successfully in vitro and were identified positive by the function of cells and immunofluorescence staining. The endothelial progenitor cells were infected directly by the recombinant adenovirus-associated virus containing β-NGF gene with an efficiency of 65.3%. β-NGF protein was detected in the culture supernatant of transfected endothelial progenitor cells, which reached a high level at 10 days after gene transfection. Furthermore, there was no β-NGF protein in the blank and empty vector groups. After transfection, the proliferative ability of endothelial progenitor cells was increased, which was significantly higher than the blank and empty vector groups (P < 0.05). But there was no difference between the latter two groups (P > 0.05). These findings suggest that recombinant adenovirus-associated virus containing β-NGF gene can be successfully transferred into rat bone marrow-derived endothelial progenitor cells and promote the proliferation of endothelial progenitor cells. 中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 Figures and Tables | References | Related Articles | Metrics

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Constructing a lentiviral vector overexpressing indoleamine 2,3-dioxygenase  He Ji-gang, Li Hong-rong, Gui Long-sheng, Li Yong-wu, Yan Dan 2015, 19 (36):  5859-5864.  doi: 10.3969/j.issn.2095-4344.2015.36.022 Abstract ( 282 )   PDF (4491KB) ( 362 )   Save BACKGROUND: Immune rejections after organ transplantation or serious adverse reactions due to immunosuppressive drugs show a lack of effective treatments and poor therapeutic outcomes. Therefore, we try to find an effective immune suprresion method in combination of the latest immunomodulatory achievements. OBJECTIVE: To construct a lentiviral vector overexpressing indoleamine 2,3-dioxygenase (IDO). METHODS: (1) The IDO gene that was successfully contructed was inserted into lentiviral packaging plasmids GV308 to construct GV308-IDO lentivirus packaging plasmids. (2) The 293T cells with 80% confluence were co-cultured with 5'LTR and 3'LTR, basic elements of lentiviral packaging auxiliary components, including Psi, cPPT, 3FLAG, TetR, IRES, WRPE, TetIIP, Ubiquitin Promoter, SV40 origin and HIV. RESULTS AND CONCLUSION: Western blot assay showed that in 10 g/L agarose gel electrophoresis, there was a target fragment at Mr 48 000. This value was consistent with the size of IDO protein. RT-PCR results showed visible IDO expression in 293T cells. These findings suggest that IDO fusion gene has been successfully reorganized in the lentiviral packaging plasmids. 中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 Figures and Tables | References | Related Articles | Metrics

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Mesenchymal stem cell transplantation for spinal cord injury: a Meta-analysis Zhao Wen-tao, Li Pan-pan, Zhang Hai-feng, Wu Na-ping, Liang Jian-fang 2015, 19 (36):  5865-5871.  doi: 10.3969/j.issn.2095-4344.2015.36.023 Abstract ( 385 )   PDF (1155KB) ( 416 )   Save BACKGROUND: In recent years, the effectiveness of stem cell transplantation in the treatment of spinal cord injury has been validated in animal models, and mesenchymal stem cell transplantation for treatment of spinal cord injury has been studied most widely. Currently, there are a number of relevant clinical studies that have shown a good prospect. OBJECTIVE: To evaluate the efficacy and safety of mesenchymal stem cell transplantation for spinal cord injury in human with a system review. METHODS: PubMed database, EMBASE database, Cochrane Library, ISI Web of knowledge, CBM database, VIP database, CNKI database and Wanfang database were searched from their start year up to July 2015 for relevant randomized clinical trials on the treatment of spinal cord injury with mesenchymal stem cell transplantation. The key words were “spinal cord injury, paraplegia, cell transplantation, transplantation,mesenchymal stem cell, bone marrow transplantation, stem cell, randomized controlled trial” in English and Chinese, respectively. RESULTS AND CONCLUSION: A total of 260 articles were retrieved, including 6 randomized clinical trials (252 cases). In the aspects of ASIA touch sensation score, overall Frankel score and daily life activity training score, the patients undergoing mesenchymal stem cell transplantation were significantly superior to those in the control group (P < 0.05). In addition, ASIA motor function score and residual urine volume were also improved in the patients undergoing mesenchymal stem cell transplantation, but there was no statistical difference (P > 0.05). Compared with the control group, low fever was more common in the patients undergoing mesechymal stem cell transplantation (P < 0.05). Another side effect was lower limb numbness, but there was no difference from the control group (P > 0.05). These findings suggest that mesenchymal stem cell transplantation has limited efficacy in the treatment of spinal cord injury and cannot induce severe complications, but there is a need for high-quality randomized controlled trials to prove the efficiency and safety of mesenchymal stem cell transplantation for the treatment of spinal cord injury. 中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 Figures and Tables | References | Related Articles | Metrics

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Stem cell-induced proliferation of myocardial cells and exercise-induced regeneration of myocardial cells Chen Guang-tao1 Zhang Lan, Ren Wen-jun 2015, 19 (36):  5872-5877.  doi: 10.3969/j.issn.2095-4344.2015.36.024 Abstract ( 290 )   PDF (889KB) ( 279 )   Save BACKGROUND: Stem cell transplantation has incomparable superiorities in the treatment of cardiovascular diseases. But stem cells have a very low efficiency to differentiate into myocardial cells spontaneously, and there are many factors influencing stem cell differentiation. OBJECTIVE: To clarify the pros and cons of different sources of stem cells in the treatment of myocardial infarction, to investigate the methods for improving the differentiation efficiency of myocardial cells, optimal differentiation conditions and mechanism underlying exercise-induced stem cell mobilization and endogenous myocardial cell regeneration. METHODS: A computer-based search of CNKI and PubMed databases was performed by the first author for articles related to stem cell therapy for myocardial infarction and stem cell differentiation into myocardial cells, exercise effect on stem cell proliferation and myocardial cell regeneration published from 1985 to 2015. The key words were “stem cells, myocardial infarction, myocardial regeneration, cardiac cell, exercise” in Chinese and English, respectively. Finally, 54 articles were included in result analysis. RESULTS AND CONCLUSION: In recent years, a variety of chemical inducers and biological components have been used in the myocardial differentiation of stem cells. Simulation of myocardial microenvironment and vascular cell growth factors are the main methods of inducing myocardial differentiation. Aerobic exercise-induced stem cell mobilization can induce ischemic cardiac angiogenesis and upregulate a variety of vascular endothelial growth factors so as to promote myocardial proliferation and repair. However, in-depth exploration is still needed in the harvesting of stem cells, transplant rejection, and regulatory mechanisms underlying the directed differentiation of stem cells into cardiomyocytes. 中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 Figures and Tables | References | Related Articles | Metrics

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Umbilical cord mesenchymal stem cells promote angiogenesis of ischemic lower limbs Li Xiao-ling, Zhu Lv-yun, Song Guang-yao 2015, 19 (36):  5878-5885.  doi: 10.3969/j.issn.2095-4344.2015.36.025 Abstract ( 170 )   PDF (987KB) ( 323 )   Save BACKGROUND: Under certain conditions, stem cells can be induced to differentiate into vascular endothelial cells, which can promote the angiogenesis of ischemic lower limbs and the establishment of effective circulation and improve distal blood supply of the ischemic limbs. OBJECTIVE: To review the biological characteristics and pro-angiogenesis mechanism of umbilical cord mesenchymal stem cells and to investigate the current status of umbilical cord mesenchymal stem cells in the repair of neuropathy and chronic wounds. METHODS: PubMed, VIP and Wanfang databases were searched for relevant articles published from 2000 to 2015 using the keywords of “stem cells transplantation, umbilical cord mesenchymal stem cell, diabetic angiopathies” in English and Chinese, respectively. RESULTS AND CONCLUSION: Compared with peripheral blood stem cells and bone marrow mesenchymal stem cells, umbilical cord mesenchymal stem cells are characterized as more widespread sources, easy collection, stronger amplification ability, no immunogenicity, and no ethical controversy, which have become ideal target and seed cells for pro-angiogenesis and gene therapy in ischemic diseases. Umbilical cord mesenchymal stem cells can differentiate into vascular endothelial cells and fibroblasts involved in wound healing. In addition, these cells can promote the production and expression of neurotrophic factors, promote nerve regeneration in ischemic tissues, and participate in tissue repair and accelerate healing of ulcers by paracrine and autocrine cytokines, anti-inflammation and immunomodulation. Therefore, umbilical cord mesenchymal stem cells have a broad prospect in the improvement of diabetic lower limb ischemia, repair of diabetic peripheral neuropathy and promotion of chronic ulcer healing. Compared with stem cell transplantation alone, umbilical cord mesenchymal stem cells transplantation combined with gene therapy can further enhance cell survival and pro-angiogenesis. 中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 Figures and Tables | References | Related Articles | Metrics

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Synovial mesenchymal stem cells for knee cartilage injury  Chen Qi, Liao Wen-bo 2015, 19 (36):  5886-5891.  doi: 10.3969/j.issn.2095-4344.2015.36.026 Abstract ( 229 )   PDF (1092KB) ( 340 )   Save BACKGROUND: Knee cartilage injury is difficult to heal, which is an urgent clinical problem to solve. OBJECTIVE: To summarize the advantages of synovial mesenchymal stem cells in the treatment of knee cartilage injury. METHODS: A computer-based search of PubMed was performed for articles related to synovial mesenchymal stem cells for knee cartilage injury using the keywords of “knee joint, cartilage, synovial membrane, tissues, injuries and repair”. A total of 48 articles were retrieved initially, and 35 articles were included in result analysis. RESULTS AND CONCLUSION: Synovial mesenchymal stem cells for treatment of knee cartilage injury can achieve more ideal outcomes. Mesenchymal stem cells inherent in the damaged tissue are the optimal seed cells for tissue repair. 中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 Figures and Tables | References | Related Articles | Metrics

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Intra-articular injection of synovial mesenchymal stem cells to treat articular cartilage injury  Yuan Xiao-long, Bi Sheng-rong, Yu Fang-yuan 2015, 19 (36):  5892-5897.  doi: 10.3969/j.issn.2095-4344.2015.36.027 Abstract ( 208 )   PDF (860KB) ( 397 )   Save BACKGROUND: Among various seed cells, synovial mesenchymal stem cells have unique advantages in the repair of articular cartilage injury. OBJECTIVE: To review the progress of synovial mesenchymal stem cells and its intra-articular injection in the treatment of articular cartilage injury. METHODS: The first author searched PubMed and CNKI by computer to retrieve articles published from January 2004 to December 2004 using the keywords of “synovial mesenchymal stem cells; intra-articular injection; cartilage repair” in English and Chinese, respectively. Finally, 57 articles were included in result analysis. RESULTS AND CONCLUSION: It is easy to isolate and culture synovial mesenchymal stem cells, which has great advantages in cartilage repair. What’s more, intra-articular injection therapy for articular cartilage injury is feasible and safe. Intra-articular injection of synovial mesenchymal stem cells is a very promising treatment for cartilage damage, but there are still many problems to be solved in the future. 中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 Figures and Tables | References | Related Articles | Metrics

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Oxidative stress and stem cell transplantation Yang Min, Wang Xin-fa, Liu Ying-xia, Xiao Jia 2015, 19 (36):  5898-5904.  doi: 10.3969/j.issn.2095-4344.2015.36.028 Abstract ( 204 )   PDF (913KB) ( 477 )   Save BACKGROUND: Stem cells with wide variety of sources have the potential of differentiation and self-renewal. In additional, autologous stem cells can avoid immune rejection after transplantation, and thus it has become one of the most promising alternative strategies for tissue/organ transplantation. However, due to the adverse environments at injured sites, such as oxidative stress and inflammation, current stem cell transplantation efficacy is relatively low. OBJECTIVE: To review the effects of oxidative stress on stem cells and on their transplantation efficiency as well as relevant mechanisms. METHODS: PubMed database was searched by the first author for relevant articles about stem cells published from 1990 to 2015. The keywords were “stem cell transplantation, stem cell, oxidative stress, molecular mechanism”. After eliminating literatures which had poor authority or similar content, 97 articles were involved in result analysis.  RESULTS AND CONCLUSION: Different types of stem cells have different basal endogenous antioxidant stress levels. Oxidative stress through multiple molecular pathways causes cell aging, apoptosis and cancer, which also can result in apoptosis of cancer cells. Stem cells can adjust endogenous antioxidant levels through multiple paths. To improve the endogenous antioxidant stress level using a variety of methods can increase stem cell transplantation efficiency and prevent stem cell cancerization due to oxidative stress, which makes the clinical application of stem cell transplantation therapy safer and more popular. 中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程 Figures and Tables | References | Related Articles | Metrics