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02 July 2010, Volume 14 Issue 27 Previous Issue    Next Issue

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Directional differentiation and identification of bone marrow mesenchymal stem cells into structural cells for heart valves Fu Jie, Jiang Ying-jiu, Jiang Rong 2010, 14 (27):  4941-4945.  doi: 10.3969/j.issn.1673-8225.2010.27.001 Abstract ( 325 )   PDF (396KB) ( 376 )   Save BACKGROUND: In the component of cardiac valve, not only endothelial cells exert effects, but also fibroblasts and smooth muscle cells participate in the construction of matrix of cardiac valve. OBJECTIVE: To establish a simple method for obtaining large quantity of endothelial cells, smooth muscle cells and fibroblasts in vitro, and identify the morphology and surface markers of the obtained cells. METHODS: Bone marrow mesenchymal stem cells (BMSCs) were isolated from rats by whole bone marrow culture method, then cultured in vitro, and proliferated. The third passage of BMSCs was treated in special induction medium for differentiation into endothelial cells, vascular smooth muscle cells and fibroblasts. The cells were detected under an inverted microscope to examine the cell morphology every day, and then the differentiated cells were measured by immunocytochemistry. RESULTS AND CONCLUSION: The cells induced from BMSCs had the characteristics of endothelial cells, smooth muscle cells and fibroblasts. Results have indicated that BMSCs could be directionally differentiated into endothelial cells, smooth muscle cells and fibroblasts in vitro. Related Articles | Metrics

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Platelet-rich plasma induced bone marrow mesenchymal stem cells combined with chemical extraction of acellular nerve for repair of sciatic nerve defect Ding Chang-rong, Yang Xuan-ying, Han Ying-qiu, Xia Chang-suo, Wang Ying-zhen 2010, 14 (27):  4946-4950.  doi: 10.3969/j.issn.1673-8225.2010.27.002 Abstract ( 240 )   PDF (641KB) ( 543 )   Save BACKGROUND: Schwann cells have not yet a large number of division and proliferationin following early peripheral nerve injury. As anatomic discontinuity, the nutritional factors through axoplasmic retrograde transport reduce sharply. The neurons are likely to die due to lack of neurotrophic factors support, so peripheral nerves cannot regenerate or show weak regeneration. OBJECTIVE: To evaluate the effects of the implantation of bone marrow mesenchymal stem cells (BMSCs) induced by the platelet-rich plasma (PRP) combined with chemical extraction of acellular nerve on repairing sciatic nerve defect. METHODS: New Zealand white rabbits were selected to establish sciatic nerve defect models, and randomly divided into four groups, namely pure chemical extracted acellular nerve group, transplantation of allogenic acellular nerve; BMSC group, transplantation of allogenic BMSCs combined with chemical extraction of allogenic acellular nerve; induced BMSC group, transplantation of the PRP-induced allogenic BMSCs combined with chemical extraction of allogenic acellular nerve; and autologous nerve group, transplantation of autologous nerve. Detection indicators include morphological observation, the target muscle recovery rate of muscle wet weight, motor nerve conduction velocity (MNCV) and the axon diameter and myelin sheath thickness. RESULTS AND CONCLUSION: Target muscle to restore the rate of muscle wet weight, MNCV, axon diameter, myelin sheath thickness and the morphology in PRP induced by BMSCs combined with chemical extraction of acellular nerve group was significantly superior to a pure chemical extracted acellular nerve group and BMSCs combined with chemical extraction of acellular nerve group, but there were similar with autologous nerve repair group. BMSCs have the function of Schwann cells in vivo after induction. BMSCs can be used as tissue-engineered peripheral nerve of the seed cells for the repair of peripheral nerve defects Related Articles | Metrics

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Experimental observation of rabbit intervertebral disc transplantation with human bone marrow mesenchymal stem cells Wang Yue-qiu, Chen Bo-hua 2010, 14 (27):  4951-4954.  doi: 10.3969/j.issn.1673-8225.2010.27.003 Abstract ( 299 )   PDF (419KB) ( 379 )   Save BACKGROUND: Numerous studies have demonstrated allogenic bone marrow mesenchymal stem cells (BMSCs) transplantation, but few reports have addressed the survival of human BMSCs in the intervertebral discs in vivo. OBJECTIVE: To observe the survival of human BMSCs in the rabbit intervertebral discs. METHODS: A total of 15 New Zealand white rabbits were selected, and each of the rabbit intervertebral disc was divided into 3 groups, blank group(L1-2), physiological saline (PS) group(L2-3), green fluorescent protein (GFP) transfection of human BMSCs transplantation group(L3-4, L4-5, L5-6). The samples in the blank group were not injected; those in the PS group were injected with 25 μL PS into nucleus pulposus; those in the human BMSCs/GFP transplantation group were injected with 25 μL human BMSCs/GFP-PS suspension(1×109/L) into nucleus pulposus. At 1, 2, 4, 6, 8 weeks after operation, the specimen was taken out. Fluorescence microscope was used to observe distributed situation and quantity.  RESULTS AND CONCLUSION: Samples were sliced into sections at 1, 2, 4, 6, 8 weeks following GFP-labeled human BMSCs transplantation. Green fluorescent cells were seen in the nucleus pulposus. The number of fluorescent cells was significantly less at 6 and 8 weeks than at 1, 2, 4 weeks following transplantation(P < 0.001). Results have suggested that within 8 weeks, the human BMSCs could be survived in the rabbit intervertebral disc. Related Articles | Metrics

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Monolayer coculture of rabbit chondrocytes and bone marrow stromal cells: Screening of culture proportion Sun Lei, Qi Hui, Chen Lei, Chen Hai-yan, Jiang Jian, Tao Jian-feng, Jie Yong-sheng, Gao Xin-sheng, Peter I. Lelkes 2010, 14 (27):  4955-4959.  doi: 10.3969/j.issn.1673-8225.2010.27.004 Abstract ( 282 )   PDF (453KB) ( 375 )   Save BACKGROUND: Bone marrow stromal cells (BMSCs) can differentiate into chondrocytes under a certain condition, but there are lack of studies concerning monolayer coculture of chondrocytes and BMSCs at present. OBJECTIVE: To identify percentage choice of coculture of BMSCs and chondrocytes, cell proliferation activity and cartilage specific protein expression rule, and to find the best time and subculture frequency during cell transplantation. METHODS: Chondrocytes and BMSCs were isolated, cultured, and divided into 5 groups following passage: chondrocyte group, coculture (chondrocytes and BMSCs at 7: 3, 5: 5, 3: 7) groups, and BMSC group. At generation 4(G4), cell morphology was observed at different passages in each group under an inverted phase contrast microscope. MTT assay was performed for testing the cell viability. The proteoglycan expression was detected by toluidine blue and alcian blue staining. Type II collagen expression was determined by immunocytochemical method. RESULTS AND CONCLUSION: Cells showed normal appearance in all the coculture groups. From G1-G3, cells had good viability, which was decreased at G4. Proteoglycan (detected by toluidine blue and alcian blue staining) and type II collagen expression were highly positively determined in the coculture groups during the second and third generations (G2 and G3). Synthetic each index showed that each index was best in the chondrocytes and BMSCs at 5: 5, 3: 7 groups. Results displayed that monolayer coculture of chondrocytes and BMSCs kept chondrocyte morphology and protein expression property. The proportion of chondrocytes and BMSCs at 5: 5 or 3: 7 was optimal for cell transplantation. Related Articles | Metrics

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Effect of human bone marrow mesenchymal stem cells on T cell activation in patients with systemic lupus erythematosus Dong Yi, Liu Lei, Xia Rui-xiang, Zeng Qing-shu, Li Qing-sheng 2010, 14 (27):  4960-4963.  doi: 10.3969/j.issn.1673-8225.2010.27.005 Abstract ( 291 )   PDF (423KB) ( 539 )   Save BACKGROUND: Bone marrow mesenchymal stem cells (BMMSCs) have immunological effects and can treat autoimmune disease. OBJECTIVE: To explore the effect of human BMMSCs on T cell activation in patients with systemic lupus erythematosus (SLE) in vitro. METHODS: BMMSCs were cultured and the 3rd passage cells were digested by trypsin. The concentration of cells was identified by flow cytometry. And MMSCs were divided into 1×108/L and 1×107/L group. Totally 10 mL peripheral blood lymphocytes (PBL) were obtained from 27 cases with SLE and incubated into BMMSCs with concentration of 1×109/L, 100 µL per well. Expression rates of CD3+ T cells CD25 (IL-2R) and CD38 were calculated by flow cytometry. RESULTS AND CONCLUSION: Compared with the control group, the expression of CD25 and CD38 were obviously suppressed in the 1×108/L group (P < 0.01). But there was no significant change in the 1×107/L group. The results demonstrated that BMMSCs inhibit T cell activation in patients with SLE in a dose-dependent manner. Related Articles | Metrics

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Construction and effects of bone marrow mesenchymal stem cells-cytosine deaminase/5-fluorocytosine suicide gene therapy system Cao Shi-bo, Meng Qing-hai, Jin Peng, Wang Hong-wei, Li Shi-fang, Dou Yi-he, Meng Cheng-dong, Yan Jing-jie 2010, 14 (27):  4964-4969.  doi: 10.3969/j.issn.1673-8225.2010.27.006 Abstract ( 144 )   PDF (601KB) ( 358 )   Save BACKGROUND: In vivo study has verified that bone marrow mesenchymal stem cells (BMSCs) can migrate and integrate in the brain as neural stem cells (NSCs). If BMSCs can be used in system pathway transplantation, it will simplify the procedures of transplantation. OBJECTIVE: To construct recombinant expression plasmid pEGFP-N3-CD containing cytosine deaminase (CD). Liposome Lipofectamine2000 was used to transfect rat BMSCs. To observe CD gene expression and suicide effects of BMSC-CD/5-fluorocytosine (FC) suicide gene therapy system on C6 glioma cells in vitro.  METHODS: The pEGFP-N3-CD plasmid was constructed, and determined by enzyme digestion and DNA sequence. BMSCs were harvested and cultured using the whole bone marrow method. The pEGFP-N3-CD plasmid was used to transfect rat BMSCs in vitro using Lipofectamine2000. CD gene protein expression of G418-resistant clones (named BMSCs-CD cells) was detected by immunocytochemistry. Apoptosis of C6 cells cultured with BMSCs-CD cells in Transwell culture system induced by 5-FC (24 hours later) in BMSCs-CD cells genetically modified to express CD was investigated by applying TUNEL, MTT and flow cytometry analysis techniques (72 hours later). RESULTS AND CONCLUSION: The pEGFP-N3-CD plasmid containing complete CD gene sequence was constructed. CD gene was transfected to BMSCs, and fully expressed in gene and protein levels. Under in vitro co-culture conditions of BMSCs-CD and C6 glioma cells, apoptotic rate of C6 glioma cells showed dose-dependent effects, and significant difference was detected as compared with control group (P < 0.01). Results have indicated that BMSCs-CD/5-FC suicide gene therapy system exhibits significant inhibitory effects on growth of C6 glioma cells. Related Articles | Metrics

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Screening of optimal labeling time and labeling concentration of rabbit bone marrow mesenchymal stem cells by 5-bromodeoxyuridine in vitro Zhu Feng1, Guo Guang-hua2, Zhan Jian-hua2, Xie An3 2010, 14 (27):  4970-4974.  doi: 10.3969/j.issn.1673-8225.2010.27.007 Abstract ( 71 )   PDF (579KB) ( 589 )   Save BACKGROUND: Currently, bone marrow mesenchymal stem cells (BMSCs) are potential cells and host cells for gene therapy. Labeling in vitro is a premise to track their survival, “homing”, growth, differentiation and so on. OBJECTIVE: To explore the optimal 5-bromodeoxyuridine (BrdU) concentration and time of labeling for rabbit BMSCs in vitro. METHODS: Rabbit BMSCs were harvested from bone marrow of purebred healthy New Zealand white rabbits. BMSCs were identified by flow cytometry and morphology. Rabbit BMSCs-labeling positive rates using BrdU at different concentrations and different incubation time were observed. RESULTS AND CONCLUSION: Flow cytometry finally confirmed that the cells we cultured were BMSCs combined with morphological observation; the growth curve of rabbit MSCs was “S” shape. The positive rate of BMSCs labeled by BrdU increased with incubation time and BrdU concentration, and more than 85%-95% BMSCs were labeled after at optimal 40 μmol/L and 72 hours. Above-mentioned results have suggested that application of BrdU labeling for BMSCs in vitro is safe and reliable. The optimal time and concentration for labeling BMSCs are 72 hours and 40 μmol/L. Related Articles | Metrics

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Recovery rate and postresuscitation differentiation of bone marrow mesenchymal stem cells at high temperature   Ke Jun-long, Xu Zhi-en, Li Hua, Chen Jing-juan 2010, 14 (27):  4975-4978.  doi: 10.3969/j.issn.1673-8225.2010.27.008 Abstract ( 129 )   PDF (487KB) ( 367 )   Save BACKGROUND: At present, temperature of recovery of bone marrow mesenchymal stem cells (BMSCs) is 37 ℃. Previous reports have confirmed that spermatogonia were recovered at a high temperature, but it remains poor understood whether BMSCs were recovered at a high temperature. OBJECTIVE: To observe the effects of high temperature on recovery rate and differentiation ability of BMSCs. METHODS: Rat BMSCs were isolated and cultured by adherence method. The fifth passage of BMSCs were cryopreserved, and then recovered at 37 ℃ and 50 ℃ in water bath. The recovery rate was compared using trypan blue staining in both groups. BMSCs were induced to differentiate into neuron-like cells by fibroblast growth factor + epidermal growth factor + ratinoic acid at 50 ℃ in water bath, and identified by immunohistochemical staining. RESULTS AND CONCLUSION: Recovery rates were respectively (82.40±0.86)% and (87.10±1.08)% in the 37 ℃ and 50 ℃ groups (P < 0.01). In the 50 ℃ group, cells presented neuron-like changes following inducer intervention, and were positive for Nestin in immunohistochemical staining. Results have suggested that the recovery rate in BMSCs was higher in 50 ℃ group compared with 37 ℃ group. Moreover, the differentiation ability was not affected. Related Articles | Metrics

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Inhibitory effects of co-culture of umbilical cord blood-derived mesenchymal stem cells and human cardiomyocytes on cardiomyocyte apoptosis Yang Shui-xiang, Huang Jing-ling 2010, 14 (27):  4979-4983.  doi: 10.3969/j.issn.1673-8225.2010.27.009 Abstract ( 122 )   PDF (531KB) ( 388 )   Save BACKGROUND: Umbilical cord blood (UCB)-derived mesenchymal stem cells (MSCs) have multi-differentiation potentials, and can repair myocardial tissue following homing or implantation into the heart. However, transplantation safety and whether the transplantation can inhibit apoptosis of human cardiomyocytes under coculture of human cardiomyocytes deserve further investigations. OBJECTIVE: To study the safety and apoptosis inhibition of the UCB-derived MSCs with co-culture on human cardiomyocytes. METHODS: Human UCB-derived MSCs were collected at the time of delivery, and treated with 5-azacytidine for 24 hours and further introduced differentiation into cardiomyocytes. Cells at passages 3-5 and induced cells were obtained to detect the telomerase activation, tumor-related gene RNA levels, chromosome karyotype G typing, cell surface antigen expression, tumor formation in nude mice, and cell apoptosis under coculture. RESULTS AND CONCLUSION: MSCs derived from UCB were differentiated into cardiomyocytes in vitro following induced by 5-azacytidine. Telomerase activity, p53, cyclin A, cdk2, β-actin, c-fos, h-TERT and c-myc expression was similar before and after induction. No abnormal chromosomal karyotypes were observed. Immunophenotype did not significantly change. Cells were negative for CD34, but positive for CD44 and CD90 (Thy-1). There was no tumor formation in nude mice. UCB-derived MSCs significantly inhibited apoptosis of human cardiomyocytes under co-culture conditions compared with cardiomyocytes (P < 0.05). Results have suggested that human UCB-derived MSCs are a valuable safe and effective source of cell-transplantation treatment, and can inhibit the apoptosis of human cardiomyocytes under co-cultured conditions. Related Articles | Metrics

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Biological characteristics, osteogenic and adipogenic differentiation potential of mesenchymal stem cells from human umbilical cord blood Zhang Ji-hua, Sun Kang, Wang Yan, Tian Shao-qi, Xia Chang-suo, Zhang Cai-long, Yu Teng-bo 2010, 14 (27):  4984-4987.  doi: 10.3969/j.issn.1673-8225.2010.27.010 Abstract ( 81 )   PDF (426KB) ( 485 )   Save BACKGROUND: Studies addressing umbilical cord blood-derived mesenchymal stem cells (UCB-MSCs) are key point and hot focus in the research of stem cells. However, few reports have addressed UCB-MSCs, and there are controversial in many aspects. OBJECTIVE: To observe the biological characteristics, osteogenic and adipogenic differentiation potential of UCB-MSCs. METHODS: MSCs were harvested and cultured from UCB at various gestational ages. The morphology and ultramicrostructure of MSCs and their growth characteristics in vitro were observed under an inverted phase contrast microscope and transmission electron microscope. The surface antigen phenotype was analyzed by flow cytometry. The osteogenic and adipogenic media were used to induce differentiation of UCB-MSCs into osteoblasts and adipocytes. The osteogenic potential was determined by alizarin red staining. Adipogenic potential was detected by oil-red O staining. RESULTS AND CONCLUSION: Under the inverted phase contrast microscope, UCB-MSCs were adherent with a fibroblast like morphology and whirlpool like growth alinement. The ultramicrostructure in transmission electron microscope showed that UCB-MSCs had a big cell nucleus, less cellular organelles and big karyoplasmic ratio. All of the growth curves of primary and subculturing UCB-MSCs presented “S” type. The third and fifth generation of MSCs showed the greatest reproductive activity. And the count of CFU-F was higher in smaller gestational age than the older. Flow cytometry showed that these cells were positive for CD29, CD44 and CD90 expression, but they failed to express hematopoictic cell surface markers, such as CD34 and CD45. When the MSCs were induced to osteogenic and adipogenic differentiation for 3 weeks, the formation of a mineral extracellular matrix and neutral lipid vacuoles detected by alkaline phosphatase staining, alizarin red staining and oil-red O staining respectively. Results have indicated that UCB-MSCs have similar morphological character, biological characteristics and cell surface markers with MSCs derived from bone marrow, both of which have great capability of proliferation and regeneration. UCB-MSCs can be induced to differentiation to osteoblasts and lipoblasts in a suitable condition in vitro. Related Articles | Metrics

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Biological security of differentiation of human umbilical cord blood-derived mesenchymal stem cells into cardiomyocytes   Huang Jing-ling, Yang Shui-xiang, Chen Yi-rong 2010, 14 (27):  4988-4992.  doi: 10.3969/j.issn.1673-8225.2010.27.011 Abstract ( 129 )   PDF (555KB) ( 326 )   Save BACKGROUND: Under the differentiation into cardiomyocytes in vitro, few studies have addressed whether human umbilical cord blood-derived mesenchymal stem cells (UCB-derived MSCs) can maintain their normal physiological properties. i.e. whether reach the biological safety standards of seed cells. OBJECTIVE: To analyze the changes of chromosome karyotype, telomerase activity and tumorigenicity under a condition of cardiomyocyte differentiated from human UCB-derived MSCs that were isolated and cultured in vitro. METHODS: Karyotypes were analyzed in UCB-derived MSCs from the third passage that was isolated and cultured in vitro using chromosome G-banding technique and samples following the seventh passage of cells differentiating into cardiomyocytes. Cell telomerase activity and cell cycle-related genes cyclinA, cdk2, C-fos, h-TERT, c-myc and p53 expression were analyzed. The nude mice carcinogenic test was performed to test the tumorigenicity of cells. RESULTS AND CONCLUSION: No abnormal changes in the chromosome karyotype and telomerase activity were detected in human UCB-derived MSCs following in vitro cardiomyocyte culture up to the seventh passage. There were no c-myc, p53, h-TERT and C-fos expressions in tested cells. Any abnormal changes such as skin nodules or doubtful focus in the nude mice could not be found. These are consistent with the National Medical Product Biology Evaluation Criterion. Related Articles | Metrics

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Effects of Notch1 gene silencing in mouse embryonic stem cells on glycogen synthase kinase-3 beta and cyclin-dependent kinase 5 expression Zhang Qiu-ying, Duan Ping, Han Xue-fei, Xu Yan, Xing Ying, Yan Wen-hai 2010, 14 (27):  4993-4996.  doi: 10.3969/j.issn.1673-8225.2010.27.012 Abstract ( 127 )   PDF (552KB) ( 372 )   Save BACKGROUND: Glycogen synthase kinase-3β (Gsk-3β) and cyclin-dependent kinase 5 (Cdk5) are phosphorylated protein kinase, and participate in differentiation and development of embryonic neuronal cells, but the precise mechanism remains unclear. OBJECTIVE: To detect the changes in genetic expression of Hes-1, PS1, Gsk-3β and Cdk5 after Notch1 gene silencing in embryonic stem cells (ESCs), thus to explore Notch1 signaling accommodative action on Gsk-3β and Cdk5 and its potential impact on early neurogenesis.      METHODS: Lipofectamine 2000 was used to transfer Notch1 small interfering RNA (siRNA) eukaryotic expression vector into mouse ESCs. Morphologic changes of cells before and after transfection were observed under inverted microscope. ESCs were collected at 48 hours after transfection. Genetic expressions of Notch1, Hes-1, PS1, Gsk-3β and Cdk5 in these cells are detected by RT-PCR. The intergroup differences were analyzed using one-way variance analysis. RESULTS AND CONCLUSION:  ①Relative expression of Notch1, Hes-1, Gsk-3β and Cdk5 in pSINsi-U6-Notch1 group was obviously lower than blank plasmid and normal control groups ( P <0.05). There were no significant differences between blank plasmid and normal control groups (P > 0.05, n=5). ②Under an inverted microscope, exogenous plasmid did not impact ESCs cloning growth, but the proliferation quantity of cells in pSINsi-U6-Notch1 group was decreased compared with blank plasmid and normal control groups. ③No significant differences in genetic expression of PS1 were detected among the three groups (P > 0.05). Decreased Notch1 gene expression in ESCs down-regulates gene expression of Gsk-3β and Cdk5. Related Articles | Metrics

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Safety of human umbilical cord mesenchymal stem cells transplanted into rats by intramuscular injection   Zhang Mei-rong, Zhang Xue-feng, Hu Jian-xia, Wang Li, Zhang Gui-zhi, Gao Hong, Miao Zhi-min, Wang Yan-gang 2010, 14 (27):  4997-5000.  doi: 10.3969/j.issn.1673-8225.2010.27.013 Abstract ( 95 )   PDF (338KB) ( 489 )   Save BACKGROUND: A few reports have addressed the safety of mesenchymal stem cells (MSCs) by intramuscular injection in China. OBJECTIVE: To observe each physiological index and pathological changes of local muscle tissue following human umbilical cord MSCs (HUMSCs) transplanted into rat muscle by intramuscular injection. METHODS: In a total of 51 male SPF Wistar rats, three served as blank controls, and the remaining was randomly divided into four groups. 2.5×109/L, 5×109/L, 1.5×1010/L HUMSCs were respectively injected into the gastrocnemius of the left lower limb of rats of low, moderate and high cell concentration groups, totally two entry sites, 0.1 mL/site. The distance between two sites was 0.5 cm. 50 g/L glucose solution was infused into rats of the solvent control group. Urine routine test, blood routine test, hematological and biochemical test and histopathologic examination were conducted at 1 day, 1, 2, 4 weeks following injection. RESULTS AND CONCLUSION: No significant effect was observed on urine routine test and histopathologic examination of liver and kidney after injection of HUMSCs. Total bilirubin levels were transient, or platelet, lactate dehydrogenase, and creatine kinase-MB levels slightly higher exceeding upper normal range caused by inflammatory reaction. High cell concentration could cause significant inflammatory reaction in the injection site of muscle tissues. Results have suggested that HUMSCs do not induce severe immunoreactivity even to xenografts by intramuscular injection owing to their low immunogenicity under suitable concentration and dose of cell transplantation. Related Articles | Metrics

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Inducing human parthenogenetic embryonic stem cells into islet-like clusters Zhou Jing, Li Jin, Lin Ge, Lu Guang-xiu 2010, 14 (27):  5001-5005.  doi: 10.3969/j.issn.1673-8225.2010.27.014 Abstract ( 120 )   PDF (525KB) ( 384 )   Save BACKGROUND: Following the successful establishment of human parthenogenetic embryonic stem (hpES) cell lines, the further functional study becomes hot spot. OBJECTIVE: To investigate the potential differentiation ability to islet-like clusters from hpES cell lines. METHODS: One self-established hpES cell line and one normal hpES cell line were obtained. We adopted a modified 5-stages protocol which based on the development rule of pancreas to induce hpES cells into islet-like clusters. RESULTS AND CONCLUSION: The terminal differentiated cells gathered into islet-like clusters. The results from immunohistochemistry and RT-PCR showed that the islet-like clusters expressed the islet specific hormones and functional markers. Insulin release test indicated that the clusters had the biochemical function of islet. The islet-like clusters derived from phES cells had the basic characteristics of islet, which would be a reliable material to treat type I diabetes mellitus. Related Articles | Metrics

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Analyses of gene sets associated with embryonic stem cells in different histological grades of ovarian cancer Ye Yun, Wang Gui-ping, Yang Xiao-qin, Yin Xi, Liang Shuang, Zheng Wen-ling, Ma Wen-li 2010, 14 (27):  5006-5009.  doi: 10.3969/j.issn.1673-8225.2010.27.015 Abstract ( 137 )   PDF (353KB) ( 846 )   Save BACKGROUND: Recent studies have found that tumor cells have the characteristics of stem cells. Therefore, the gene regulation network that controls the function of stem cells may play important roles in some tumors. OBJECTIVE: To study the molecular trait of ovary cancer in different grades, and to obtain the expression of gene sets associated with embryonic stem cells in ovary cancer. METHODS: Gene expression profile data of ovarian cancer (accession number GSE2109) were obtained from Gene Expression Omnibus (GEO) database of National Center for Biotechnology Information. Ovarian cancer samples served as study materials. Samples without clinical data were excluded and then separated into two groups (high differentiation and low differentiation) according to histological grade. Expression value of samples was obtained after raw profile data were normalized and quality control by dChip. Matrix of gene expression was obtained. The enrichment of gene sets, biological process and KEGG pathway were analyzed in ovarian cancer of various differentiations by GSEA software. RESULTS AND CONCLUSION: A total of 13 gene sets associated with embryonic stem cells, 9 upregulated in poorly differentiated tumors, whereas 4 associated with PRC2 target downregulated. These suggested that gene sets associated with embryonic stem cells upregulated in poorly differentiated tumors. And pathways related cell cycle, cell division and DNA replication also enrich in poorly differentiated tumors. Gene expression profile revealed poorly differentiated ovary tumors possess the similar molecular trait with embryonic stem cells. These analyses may benefit further investigations of early diagnosis and treatment of ovarian cancer. Related Articles | Metrics

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Effects of intravenous administration of human umbilical cord blood mononuclear cells on cardiomyocyte apoptosis and apoptosis genes bcl-2 and bax in rabbits after myocardial infarction   Wang Tao, Yu Guo-long, Zhou Bo, Pan Wei, Qing Li-qiong 2010, 14 (27):  5010-5014.  doi: 10.3969/j.issn.1673-8225.2010.27.016 Abstract ( 93 )   PDF (539KB) ( 443 )   Save BACKGROUND: The previous studies showed that intra-coronary or intra-myocadial intransplantation of human umbilical cord blood mononuclear cells (HUCBC) could promote angiogenesis, and reduce myocardial infarcted area associated with improvement of cardiac function. But, there is no study about the effect of human umbilical cord blood cells on cardiomyocyte apoptosis. OBJECTIVE: To observe the effects of intravenous administration of HUCBC on cardiomyocyte apoptosis and the expressions of Bcl-2 and Bax proteins after acute myocardial infarction (AMI). METHODS：A total of 45 adult rabbits were divided into three groups randomly: transplantation group with 15 rabbits was intravenously administrated with 500 μL HUCBC labeled with 2×107 bromodexyuridine (BrdU) at 24 hours after AMI: control group with 15 rabbits was intravenously administrated with 500 μL saline at 24 hours after AMI；sham operation group with 15 rabbits, not receiving ligation of left anterior descending branch, was intravenously administrated with 500 μL saline at 24 hours after operation. Cardiac functions were performed by echocardiography at 1, 2 and 4 weeks after transplantation. Myocardial tissue sections were observed using hematoxylin-eosin staining to measure pathological changes in myocardium at 4 weeks following transplantation. The apoptotic cells were detected in the myocardium by terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL). BrdU-positive cells and the expression of Bcl-2 and Bax proteins were identified in the myocardium by immunohistochemical method. RESULTS AND CONCLUSION: ①Compared with control group, cardiac functions were significantly improved in the transplantation group. ②BrdU-positive cells were found surrounding infarct site in the transplantation group. ③Compared with the control group, Bcl-2 protein levels were significantly increased, but Bax protein levels were significantly decreased in the transplantation group. ④Number of apoptotic cardiomyocytes was significantly less in the transplantation group than the control group at 4 weeks following transplantation. These verified that HUCBCs by intravenous administration migrate into the peri-infarcted area, and regulate the expression of Bcl-2 and Bax proteins, and inhibit cardiomyocyte apoptosis in the peri-infarcted area, and finally improve cardiac function following myocardial infarction. Related Articles | Metrics

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Application of frozen single platelet transfusion in peripheral blood stem cell transplantation: Comparison with fresh single platelet transfusion Yang Xiao-shun, Xie Lin, Zhu Hong-jie, Li Lin-hua, Wang Gui-hua 2010, 14 (27):  5015-5017.  doi: 10.3969/j.issn.1673-8225.2010.27.017 Abstract ( 113 )   PDF (308KB) ( 456 )   Save BACKGROUND: Because voluntary blood donation in China leads to blood shortage, a large number of fresh single platelets are sometimes difficult to guarantee. Frozen single platelets have immediate hemostatic effect for patients who are in the low state platelets with bleeding. OBJECTIVE: To study the effect of frozen single platelet transfusion on peripheral blood stem cell transplantation. METHODS: A total of 44 patients with hematopathy or lymphoma were subjected to peripheral blood stem cell transplantation, and platelet was less than 40×109 /L. These patients were randomly assigned to frozen single platelets group and fresh single platelets group (n = 22). At 0-14 days following transplantation, patients in the fresh single platelets group were directly infused with fresh single platelets following leukocyte filtration. Homotype frozen single platelets were obtained from patients in the frozen single platelets group, and then placed in a water bath at 37 ?C for rapid thawing. Following filtration using leukocyte filter, infusion was conducted within 40 minutes; once every 3 days,  10 U/time, for totally 3-16 times. The time of infusion depended on bleeding of patients. RESULTS AND CONCLUSION: No significant difference in platelet and four items of blood coagulation was detected between fresh single platelets group and frozen single platelets group at 24 hours following infusion (P > 0.05). Blood platelet count was significantly decreased at 48 hours following infusion (P < 0.01). Bleeding time, prothrombin time, activated partial thromboplastin time and platelet count were significantly reduced at 72 hours following infusion ( P < 0.05 or P < 0.01). There were no significant differences in thrombin time and fibrinogen (P > 0.05). It is suggested that the time of frozen single platelet transfusion to prevent bleeding in peripheral blood stem cell transplantation should change into every 2 days. Related Articles | Metrics

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Effects of cardiac tissue mass cultured ex vivo on migration of mesenchymal stem cells in myocardial infarction rats Wang Ding-qiong, Shi Ruo-fei, Zhang Xiao-gang, Liao Li-qiang 2010, 14 (27):  5018-5022.  doi: 10.3969/j.issn.1673-8225.2010.27.018 Abstract ( 69 )   PDF (509KB) ( 395 )   Save BACKGROUND: Mesenchymal stem cells (MSCs) have the potential to improve cardiac function, but the mechanisms of MSCs homing to heart post-implantation are not completely clear. OBJECTIVE: To investigate the role and possible mechanisms of cardiac tissue mass cultured ex vivo which isolated from myocardial infarction rats on MSC migration.  METHODS: Rat models of myocardial infarction were established and cardiac tissue masses were cultured. MSCs were isolated from bone marrow by density gradient centrifugation and adherence screening method. A total of 24 Sprague Dawley rats were randomly assigned to myocardial infarction, sham operation and normal control groups. The rats in the sham operation group were only threaded without deligation. The rats in the normal control group were left intact. MSCs were labeled with DAPI. A transwell model was used to co-culture for 48 hours. The number of migrating cells was counted under a fluorescence microscope. Quantitative cell count was conducted for CD34/CD44 on MSCs that were tested using immunocytochemistry staining under a fluorescence microscope. Immunofluorescent staining was used to evaluate CXCR4 expression on MSCs. Immunohistochemistry staining was used to detect expression of stromal cell-derived factor-1 on cardiac sections and mean absorbance was analyzed. RESULTS AND CONCLUSION: Number of migrating MSCs in myocardial infarction group was significantly greater compared with sham operation group (P < 0.05). No migrating cells were detectable in the normal control group. Immunocytochemical staining results have shown that migrating cells were positive for CD44, but negative for CD34, which were consistent with MSC characteristics. The membrane receptor CXCR4 was positively expressed on the migrating cells. Stromal cell-derived factor-1-positive expression was observed on cardiac tissue sections both in myocardial infarction and sham operation groups, rather than in normal control group. Mean absorbance of stromal cell-derived factor-1 was significantly higher in the myocardial infarction group than in the other groups (P < 0.05). Results have indicated that cardiac tissue post myocardial infarction can promote migration of MSCs, which might be associated with effects of stromal cell-derived factor-1-CXCR4 axis. Related Articles | Metrics

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Growth and proliferation of bone marrow mesenchymal stem cells through in vitro culture of Zinc, basic fibroblast growth factor alone or their combination   Ren Li-jie, Wang Yang, Han Man-fu 2010, 14 (27):  5023-5027.  doi: 10.3969/j.issn.1673-8225.2010.27.019 Abstract ( 84 )   PDF (597KB) ( 327 )   Save BACKGROUND: It is the hot spot of neuroscience research addressing how to amplify a great quantity of bone marrow mesenchymal stem cells (BMSCs) in vitro by a convenient and effective way. OBJECTIVE: To study the effects of Zinc and basic fibroblast growth factor (bFGF) alone or in combination on the growth and proliferation of BMSCs in vitro, and to search an effective method of rapidly amplifying a great quantity of BMSCs. METHODS: Rat BMSCs were cultured in vitro. This study was composed of control group (without any influential factor), Zinc group, bFGF group, Zinc + bFGF groups, which was respectively treated with Zinc and bFGF alone or in combination. Cell growth curve was observed. Cell survival and proliferation were observed using MTT assay. Cell cycle was detected using flow cytometer, and cell surface antigen was identified. RESULTS AND CONCLUSION: Cell growth curve, MTT assay and flow cytometer results have shown that cell reproductive activity reached a peak at 3-5 days, and Zinc + bFGF group showed the strongest ability (P < 0.05). At day 7, cell reproductive activity was decreased in the control group, bFGF group and Zinc group, and only cells in the Zine + bFGF group exhibited a strong reproductive activity (P < 0.05). The percentage of cells in S+G2+M phase was greatest in the Zinc + bFGF group, and significant differences were found as compared with other three groups (P < 0.01). Results have verified that Zinc and bFGF can contribute to BMSC proliferation in vitro. Combination of Zinc and bFGF exhibits strongest effects on promoting proliferation, and can be an optimal culture condition for BMSCs in vitro. Related Articles | Metrics

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Intravenous transplantation of glial cell-derived neurotrophic factor gene modified CD34+ cells for treating cerebral infarction in spontaneous hypertensive rats Ou Ya-li, Yu Guo-long, Yang Tian-lun, Fang Li, Hu Ke 2010, 14 (27):  5028-5032.  doi: 10.3969/j.issn.1673-8225.2010.27.020 Abstract ( 73 )   PDF (532KB) ( 339 )   Save BACKGROUND: Previous studies have confirmed that direct administration of glial cell-derived neurotrophic factor (GDNF) or viral vector carrying GDNF gene can significantly reduce cerebral infarction volume, promote outcomes of the recovery of nerve function, but the therapeutic method is invasive and limits in clinic. OBJECTIVE: To observe the outcomes of intravenous transplantation of human umbilical cord blood (hUCB) CD34+ cells transfected with GDNF gene in spontaneous hypertensive rats with cerebral infarction and to explore its mechanism. METHODS: CD34+ cells isolated from hUCB were transfected by the pEGFP-GDNF plasmid and pEGFP blank vector to prepare pEGFP-GDNF-CD34+ and pEGFP-CD34+ cells. Sixty adult male spontaneous hypertensive rats with middle cerebral artery occlusion (MCAO) were randomly divided into three groups: pEGFP-GDNF-CD34+ cell group, pEGFP-CD34+ cell group and saline group. Neurological functional measurements were performed using the modified neurological severity score. Quantitative histological determinations of infarct volume were performed using standard TTC staining and quantitative image analysis. The GDNF level in the cell culture or the cerebral tissue was measured by enzyme linked immunosorbent assay. The survival and migration of green fluorescent protein (GFP)-labeled CD34+ cells and the expression of astrocytic marker-glial fibriliary acidic protein (GFAP) and the neuron marker-neuronal nuclei (NeuN) were detected by immunohistochemical and fluorescent staining. RESULTS AND CONCLUSION: Intravenous transplantation of pEGFP-GDNF-CD34+ cells migrated into ischemic-injured cerebral hemisphere, survived and differentiated into neural cells in ischemic region of spontaneous hypertensive rats, and promoted the recovery of nerve function. GDNF gene-transfected CD34+ cells survived in the brain tissue and differentiated into neural cells, and its neurological protection effect was superior to CD34+ cells. The increased GDNF level in cerebral tissue was one of possible mechanisms responsible for focal cerebral ischemia in spontaneous hypertensive rats after transplantation of between GDNF gene modified CD34+ cells and CD34+ cells. Related Articles | Metrics

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Differentiation of mesencephalic neural stem cells induced by glial cell line-derived neurotrophic factor and interleukin-1 beta in vitro   Ding Ji-gu, Ding Wen-jie, Li Guang 2010, 14 (27):  5033-5036.  doi: 10.3969/j.issn.1673-8225.2010.27.021 Abstract ( 99 )   PDF (469KB) ( 552 )   Save BACKGROUND: During neural stem cell transplantation in the treatment of Parkinson’s disease, number of transplanted cells and differentiation ratio of dopaminergic neurons must be resolved. Effective in vitro proliferation of neural stem cells and large amount of directed differentiation of dopaminergic neurons are the key to solve above-mentioned problems. OBJECTIVE: To investigate the differentiation of neural stem cells into dopaminergic neurons during induction of glial cell line-derived neurotrophic factor (GDNF) and interleukin-1β in vitro. METHODS: Mesencephalon was isolated from mouse embryo ventral part following 12 days of pregnancy, and made into single cell suspension using trypsin digestion and mechanical trituration. Following centrifugation, neurospheres were subcultured by the mechanical method for 5-7 days, and then divided into groups to induce for 10-12 days. When 80% cells migrated from neurospheres differentiated into single cells, immunocytochemical identification and flow cytometry detection were utilized to determine the rate of tyrosine hydroxylase positive cells.  RESULTS AND CONCLUSION: Neurospheres expressed nestin antigen, could differentiate into neuron specific enolase and glial fibral acidic protein-positive cells. Glial cell line-derived neurotrophic factor (GDNF) and interleukin-1β could significantly elevate the proportion of the differentiation of mesencephalic neural stem cells into tyrosine hydroxylase-positive neurons. The proportion was higher in the GDNF group, interleukin-1β group and GDNF + interleukin-1β group compared with the blank control group. In particular, the effect of GDNF + interleukin-1β was more significant. Results indicated that GDNF and interleukin-1β can obviously promote the differentiation of mesencephalic neural stem cells into enough number, mature morphology and mature function of dopamine neurons. Related Articles | Metrics

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Co-culture bio-induction of bone marrow mesenchymal stem cells and chondrocytes versus transforming growth factor beta 1 chemical induction Chen Zong-xiong, Liu Xiao-qiang, Wang Wan-zong 2010, 14 (27):  5037-5040.  doi: 10.3969/j.issn.1673-8225.2010.27.022 Abstract ( 73 )   PDF (430KB) ( 634 )   Save BACKGROUND: Bone marrow mesenchymal stem cells (BMSCs) can differentiate into chondrocytes-like cells by factor induction. However, induced cells difficultly form mature chondrocytes in an organism, and still have the ability of secreting cartilage matrix, anti-pressure and anti-friction. OBJECTIVE: To compare and analyze the effect of the induction of BMSCs by co-culture BMSCs and chondrocytes induction and transforming growth factor (TGF)-β1 induction. METHODS: Chondrocytes and the third passage BMSCs were harvested from Sprague Dawley rats, and incubated in Transwell co-culture system at 1: 2, 1: 1 and 2: 1. Simultaneously, TGF-β1 induction group served as a control. Cell proliferation and matrix synthesis would be observed under a phase contrast microscopy. MTT assay, glucose amino glycan (GAG) content detection and Western Blot assay were adopted to check the expression of type Ⅱ collagen gene. RESULTS AND CONCLUSION: The result of BMSCs and chondrocytes (1: 2) induction was quite the same as that of 10 μg/L TGF-β1 induction, but as the increasing percentage of chondrocytes in induction, the inductive result obviously has an advantage over TGF-β1 induction. When induced cells come to some rate, the result would not change, which proved that co-culture BMSCs and chondrocytes can induce BMSC transformation into chondrocytes. The induction result was positively related to the percentage of chondrocytes at the beginning; when chondrocytes come to some rate, the result of BMSCs induction does not significantly change. Above-mentioned results have shown that BMSC induction into chondrocytes exists saturation. Related Articles | Metrics

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Expression of alpha fetoprotein and albumin in bone marrow mesenchymal stem cells induced by hepatocyte growth factor   Zhang Shu-qin, Ye Dong-xia, Liu Shu-rong 2010, 14 (27):  5041-5045.  doi: 10.3969/j.issn.1673-8225.2010.27.023 Abstract ( 163 )   PDF (538KB) ( 476 )   Save BACKGROUND: Reports have addressed that induction of bone marrow mesenchymal stem cells (BMSCs) into hepatocyte-like cells in rats, mice and human. Alpha fetoprotein and albumin are specific markers of hepatocyte line. OBJECTIVE: To verify induction effects of hepatocyte growth factor on differentiation of BMSCs. METHODS: Human BMSCs were isolated and purified by density gradient centrifugation and adherent culture. BMSCs were cultured in low glucose-DMEM containing 20 μg/L hepatocyte growth factor and 10 μg/L basic fibroblast growth factor in vitro. The same passage of cells in the control group was selected and cultured in routine medium. The expression of alpha-fetoprotein and albumin was detected by immunohistochemistry at 7, 14, 21 and 28 days. The expression of albumin mRNA was detected by RT-PCR. RESULTS AND CONCLUSION: BMSCs became epithelia-shaped cells in the induction group. Alpha fetoprotein-positive rate was 81.5% at day 7 in the induction group. With prolonged time, alpha fetoprotein-positive expression became weak, whereas albumin-positive rate and albumin mRNA expression became strong. Albumin-positive expression rate was 91.2% by 28 days. In the control group, no albumin, albumin mRNA and alpha fetoprotein expression was determined. Results have verified that hepatocyte growth factor can induce BMSCs expressing alpha fetoprotein and albumin. Related Articles | Metrics

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Ex vivo expanded human umbilical cord blood mononuclear cells and hematopoietic and immune reconstitution with cytokine combination in NOD/SCID mice Xu Li, Lin Jin-ying, Mao Ping 2010, 14 (27):  5046-5049.  doi: 10.3969/j.issn.1673-8225.2010.27.024 Abstract ( 89 )   PDF (374KB) ( 404 )   Save BACKGROUND: In vitro amplification of cytokine-mediated cord blood hematopoietic cells can solve the lack of quantity during cord blood transplantation. OBJECTIVE: To explore the effect of the proper combination cytokines without stroma on the expansion of the mononuclear cells from umbilical cord blood in vitro, and to evaluate the effect of stem cell factor + FLT3 ligand + thrombopoietin on the hematopoietic and immune reconstitution. METHODS: Human fresh cord blood mononuclear cells were cultured in serum-free and stroma-free medium containing different combination of cytokines for 7 days. According to the different combination of cytokines, our experiment was divided into groups. Mononuclear cells from cord blood were expanded for 7 days in culture containing stem cell factor + FLT3 ligand + thrombopoietin, and the expanded cells were infused into sublethally irradiated NOD/SCID mice. At days 0 and 7, numbers of cord blood CD34+ cells and CD34+CXCR4+, CD34+CD49d+, CD34+CD62L+ cells were assayed. Six weeks after transplantation, the human cells in mice were assessed by flow cytometry and PCR. RESULTS AND CONCLUSION: Human CD45+ cells could be detected in the recipients 6 weeks after infusion. Detection rate of human specific genes and the survival rate of NOD/SCID mice implanted with expanded cells were higher than mice transplanted with fresh sample and saline (P < 0.05). The human cells of medullary system cells (CD33+), T lymphocytes (CD4+), B lymphocytes (CD19+) and hemopoietic stem cells (CD34+) were detected in bone marrow of NOD/SCID mice. Results indicated that the ex vivo expansion of umbilical cord blood mononuclear cells using the combination of 3 cytokines (stem cell factor + FLT3 ligand + thrombopoietin) is sufficient for clinical application. The expanded cord blood mononuclear cells can insert and reconstruct hematopoietic and immune functions of NOD/SCID mice. Related Articles | Metrics

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Angelica effects on proliferation and differentiation of neural stem cells from neonatal rats with intrauterin hypoxia Chen Xu-dong, Zhao Si-min, Hua Xin-yu, Yu Hong 2010, 14 (27):  5050-5053.  doi: 10.3969/j.issn.1673-8225.2010.27.025 Abstract ( 89 )   PDF (425KB) ( 625 )   Save BACKGROUND: Previous studies have found that intrauterin hypoxia could stimulate proliferation of neural stem cells (NSCs) of neonatal rats. The proliferation reached a peak during 6-hours hypoxia; proliferation also expressed in 9 hours, but the ability began to decline, but the neural stem cells showed necrosis or apoptosis when hypoxia up to 12 hours. However, what effect will have on neural stem cells with the time and day extension? OBJECTIVE: To study the effect of intrauterin hypoxia on the proliferation and differentiation of NSCs from neonatal rats and the protective role of angelica injection on NSCs under hypoxia. METHODS: Pregnant Sprague Dawley rats were randomly divided into control group, hypoxia group and angelica group. Pregnant rats from angelica group and hypoxia group were placed in the three-gas incubator from the beginning of pregnant 14th days to make the injured brain neonatal rat model. One hour ago, the pregnant rats of the two groups received injections respectively angelica injection and normal saline through the caudal vein of rats. Hypoxia was not performed in the control group. The remaining was identical to hypoxia group. Their brain tissues of neonatal rats were immediately taken out after childbirth. The expression of glial fibrillary acidic protein (GFAP) and neurone specific enolase (NSE) of neonatal rats was studied with immunohistochemistry and the results were analyzed by image processing system. RESULTS AND CONCLUSION: ①Expression of GFAP-positive cells in the hippocampus of neonatal rats in the hypoxia group was higher than control group. However, expression of NSE-positive cells was less in the hypoxia group than in the control group. ②Expression of GFAP-positive cells in the hippocampus of neonatal rats was less in the angelica group than in the hypoxia group, whereas expression of NSE-positive cells was higher in the angelica group than in the control group. Results have indicated that hypoxia could stimulate the proliferation of NSCs of neonatal rats and differentiation of NSCs into glial cells. Meanwhile, the number of neuron in hippocampus CA3 area was decreased. The ability of proliferation and differentiation of NSCs into glial cells after hypoxia was attenuated by angelica injection. At the same time, angelica injection could relieve neuron decrement. These suggested that angelica injection has a certain protective effect on nervous system of neonatal rats with intrauterine hypoxia. Related Articles | Metrics

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Differentiation of rat bone marrow stromal cells into neural stem cells induced by baicalin in vitro Zhang Lei, Zhang Yuan, Ni Hong-xia, Chen Li-mei 2010, 14 (27):  5054-5057.  doi: 10.3969/j.issn.1673-8225.2010.27.026 Abstract ( 83 )   PDF (471KB) ( 367 )   Save BACKGROUND: The existing experimental results show that bone marrow stromal cells have enormous potential of differentiating into bone, neural stem cells and hematopoietic stem cells, while baicalin can induce cell differentiation. OBJECTIVE: To study the possibility that induced the differentiation of bone marrow stromal cells into neural stem cells in vitro by baicalin. METHODS: Bone marrow stromal cells were harvested from Wistar rats, cultured and assigned to experimental and control groups. That in the control group was left intact. BMSCs in the experimental group were induced by baicalin (350-400 μmol/L). Both groups had the same training environment. Well grown cells following 6 consecutive days were detected using Western blot assay and reverse transcriptase-polymerase chain reaction (RT-PCR). RESULTS AND CONCLUSION: After induced by baicalin for 7 days, bone marrow stromal cells formed the more typical form of neural cells. Before induction, bone marrow stromal cells did not express neural cell marker protein mRNA and protein. After induced for 6 days, neural cells expressed marker protein and mRNA. Control group was negative for marker protein and mRNA. These indicated that baicalin may induce the differentiation of adult rat bone marrow stromal cells into neuron-like cell in vitro. Related Articles | Metrics

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Effects of lycium barbarum polysaccharides on proliferation and differentiation of rat bone marrow mesenchymal stem cells into cells of endothelial lineage   Lin Zhuang-lin, Lu Wen-yue, Chen Ping, Zheng Hui-zhen 2010, 14 (27):  5058-5061.  doi: 10.3969/j.issn.1673-8225.2010.27.027 Abstract ( 90 )   PDF (515KB) ( 711 )   Save BACKGROUND: Lycium barbarum polysaccharides (LBP) can promote spermatogenic cell proliferation and induce the differentiation of rat bone marrow messenchymal stem cells (BMSCs) and human umbilical cord blood messenchymal stem cells into neuron-like cells. But, it is unknown that the effects of LBP on biological characteristics of BMSCs, which deserves further study. OBJECTIVE: To investigate the effects of LBP on proliferation and endothelial differentiation of rat BMSCs. METHODS: BMSCs were separated from Sprague Dawley rats by gradient centrifugation. Cell proliferation rate was analyzed by MTT assay. Rat BMSCs were induced to differentiate into endothelial lineage cells by HG-DMEN plus EGM-2. These cells were identified as endothelial cells with positive factor Ⅷ and Ulex europaeus agglutinin expression. The effects of LBP on proliferation and differentiation of rat BMSCs into endothelial lineage cells were analyzed by adding LBP or not in the medium. RESULTS AND CONCLUSION: LBP in medium did not change the cellular morphology and did not affect the expression of cell surface marker CD29, CD45 or CD106 and with negative factor Ⅷ. However, LBP speeded up the proliferation of rat BMSCs and delayed its peak proliferation time. When rat BMSCs were cultured in endothelial induction medium containing EGM-2 or endothelial induction medium + LBP. In the induction group without LBP, rat BMSCs had changed into endothelial-like cells after 10 days of induction. However, these induced by medium containing LBP should change into endothelial-like cells by 12 days. These indicated that LBP can delay the differentiation of EGM-2-induced rat BMSCs into endothelial lineage cells. Related Articles | Metrics

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Anti-proliferation effect of Ponicidin on leukemia HL-60 cells Liu Xiao-dan1, Liu Wen-da2, Wang Chun-zhi, Xu Yan, Lin Dong-jun, Liu Pei-qing, Huang He-qing, Wu Chuan-bin, Xiao Ruo-zhi,Huang Ren-wei, Liu Jia-jun 2010, 14 (27):  5062-5066.  doi: 10.3969/j.issn.1673-8225.2010.27.028 Abstract ( 104 )   PDF (502KB) ( 471 )   Save BACKGROUND: Recent studies have shown that Ponicidin (PON) plays a significant role in anti-cancer effect via inducing apoptosis in various human cancer cell lines. At present, few reports have addressed the PON effects on leukemia cell lines HL-60 cells. OBJECTIVE: To investigate the anti-proliferation effect of PON on leukemic HL-60 cells in vitro and its mechanisms of action. METHODS: HL-60 cells in culture medium in vitro were given different concentrations of PON (10-50 μmol/L) for 24, 48 and 72 hours. The inhibitory rates were measured by MTT assay, and cell cycle was detected by flow cytometry. Expressions of Caspase-3 and poly(ADP-ribose) polymerase (PARP) as well as cell cycle modulator P16 and P21 were detected by Western blot assay. RESULTS AND CONCLUSION: PON could remarkably inhibit the growth of HL-60 cells and cause apoptosis; the suppression was in time- and dose-dependent manners. Flow cytometry detection revealed that the cells were mainly arrested in G0/G1 phase and the subG1 cells were also found clearly in the histogram following 50 μmol/L PON treatment. Western blot results showed cleavage of the Caspase-3 zymogen protein (Mr32 000), with the appearance of its Mr 20 000 subunit, and a cleaved Mr 89 000 fragment of PARP after the cells were treated by different concentrations of PON. Western blot analysis also revealed that P21 and P16 expression was gradually up-regulated following 50 μmol/L PON treatment. Results have indicated that PON demonstrates in vitro anti-leukemia effect in HL-60 cells mainly associated with cell cycle arrest and apoptosis. Up-regulation of the expression of P16 and P21 as well as activation of Caspase-3 may be one of its most important mechanisms of inducing HL-60 cell inhibition in G0/G1 phase or cell apoptosis. Related Articles | Metrics

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Isolation, culture and identification of adipose tissue-derived mesenchymal stem cells Yao Su-yan, Yang Yi-yong, Qin Shu-jian, Zheng De-yu 2010, 14 (27):  5067-5070.  doi: 10.3969/j.issn.1673-8225.2010.27.029 Abstract ( 191 )   PDF (477KB) ( 854 )   Save BACKGROUND: Previous studies have demonstrated that there are similar mesenchymal stem cells in adipose tissue as bone marrow mesenchymal stem cells (BMSCs). Adipose tissue can be obtained from lipid absorption of cosmetology, and is characterized by abundant source and convenient collection, which shows great value in clinical application. OBJECTIVE: To isolate and culture human adipose tissue-derived mesenchymal stem cells (ADMSCs) and to measure its biological properties. METHODS: Human ADMSCs were isolated by centrifugation and cultured in DMEM medium containing 10% fetal bovine serum. The morphology of cells was observed, and cell growth curve was drawn. Cell surface antigens were measured using flow cytometry. RESULTS AND CONCLUSION: ADMSCs obtained by digestion and centrifugation were similar in size, spindle-shaped or star-shaped fibroblasts-liked cells. Cell growth curve analysis indicated that ADMSCs were in the exponential stage at 5 days following incubation. The number of ADMSCs was reduced at 9 days. Flow cytometry analysis showed that more than 90% cells were positive for CD29, CD45 and CD105. These indicated that human ADMSCs isolated in vitro grew stably and can be used as tissue engineered seed cells. Related Articles | Metrics

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Construction of human sTRAIL vector and its expression in dermis-derived mesenchymal stem cells  Miao Dian-nan, Liang Yu-jia, Yan Guo-he, Dong Shi-wu, Dai Xiao-tian, Xiong Wei 2010, 14 (27):  5071-5074.  doi: 10.3969/j.issn.1673-8225.2010.27.030 Abstract ( 130 )   PDF (446KB) ( 311 )   Save BACKGROUND: Based on the antineoplastic features of tumour necrosis factor (TNF) soluble related apoptosis inducing ligand (sTRAIL), sTRAIL gene was cloned and its eukaryotic expression vector was constructed, which can provide a foundation for apoptotic study of tumour cells. OBJECTIVE: To construct pEGFP-N1/sTRAIL eukaryotic expression vector and express it into dermis-derived mesenchymal stem cells (dMSCs). METHODS: cDNA fragment encoding human sTRAIL gene were amplified by reverse transcriptase-polymerase chain reaction (RT-PCR) with the total mRNA isolated from human placenta tissue as template. The PCR amplified fragment of sTRAIL gene was first cloned into Pmd18-T vector, and then subcloned into the recombinant eukaryotic expression vector pEGFP-N1 to construct pEGFP-N1/sTRAIL plasmid after sequencing. Subsequently, plasmid DNA of pEGFP-N1/sTRAIL was transfected into dMSCs with the help of Fugene 6 transfection reagent. The growth of dMSCs and expression of transfected sTRAIL in dMSCs were observed under an inverted microscope. The dMSCs were identified by RT-PCR. RESULTS AND CONCLUSION: cDNA sequence encoding human sTRAIL gene was successfully cloned, and recombinant eukaryotic expression vector pEGFP-N1/sTRAIL was constructed. The expression of sTRAIL in dMSCs was obtained with the transfecton of pEGFP-N1/sTRAIL plasmid. Under an inverted microscope, the transfected DMSCs could be seen grown well. cDNA sequence encoding human sTRAIL gene was successfully cloned into pEGFP-N1. The transient expression of sTRAIL gene in dMSCs has been realized. Related Articles | Metrics

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Human embryonic stem cells culture and lung regeneration Wang Lei, Zhong Hong 2010, 14 (27):  5077-5080.  doi: 10.3969/j.issn.1673-8225.2010.27.032 Abstract ( 181 )   PDF (1KB) ( 378 )   Save BACKGROUND: In the field of pulmonary research, studies using human embryonic stem cells have succeeded in generating enriched cultures of type II pneumocytes in vitro. On account of their potential of indefinite proliferation in vitro, embryonic stem cells could be a source of an unlimited supply of cells that are available for transplantation and for use in gene therapy. OBJECTIVE: To review recent advance on human embryonic stem cell culture and lung regeneration. METHODS: We retrieved related literatures of China National Knowledge Infrastructure with the key words of “human embryonic stem cells, lung regeneration” from January 2000 to January 2010 in Chinese by computer. We then retrieved related literature of PubMed with the key words of “human embryonic stem cells, lung regeneration” from January 1990 to January 2010 in English by computer. A total of 396 articles were searched. RESULTS AND CONCLUSION: Because lung tissues have a complex structure and little capacity of regeneration, it is difficult to cure for lung diseases. At present, it is known that human embryonic stem cells are able to differentiate into lung tissue cells directionally; human embryonic stem cells are engrafted into the injured lung tissues, and induced to differentiate into alveolar epithelial cells, and further develop alveolar tissue. This is a promised therapeutic tool, but it remains in the basal research stage. Related Articles | Metrics

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Lung stem cells and the markers: Source localization and labeled difficulty Jiang Yi-yao, Du Zhen-zong, Wang Hai-yong, Zhang Zhang 2010, 14 (27):  5081-5084.  doi: 10.3969/j.issn.1673-8225.2010.27.033 Abstract ( 122 )   PDF (571KB) ( 452 )   Save BACKGROUND: In the definite condition, lung stem cells can differentiate into the functional lung tissue. In recent years, the research about the origin of lung stem cells and its markers has gained homologous progress. OBJECTIVE: To review the source of lung stem cells in the perspective of endogenous and exogenous lung stem cells, and to summarize the possible lung stem cell markers in representation of Sca-1, ABCG2/Bcrp1, residual cell marker molecules, cell surface molecules, cytokeratin, and CCSP. METHODS: A computer-based online search of China Journal Full-text Database and Pubmed was performed for articles and reviews in Chinese and English. The key words were “lung stem cells, markers”. A total of 79 articles were retrieved, and screened following reading titles and abstracts. Literatures about lung stem cells and markers were included. Repetitive articles with poor quality were excluded. Finally, 31 literatures were included. RESULTS AND CONCLUSION: The complexity of the lung tissue determined that the difficulty which was finding out the origin of lung stem cells and its markers. Currently, there are still many further explorations of the unknown fields. With the development of the experimental technique, there will be more and more new discovery. Lung stem cells will have a broad prospect in the clinical application. Related Articles | Metrics

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Neural stem cells and inner ear hearing loss and restoration Yi Tian-hua, Dong Ming-min 2010, 14 (27):  5085-5089.  doi: 10.3969/j.issn.1673-8225.2010.27.034 Abstract ( 86 )   PDF (637KB) ( 444 )   Save BACKGROUND: There are numerous studies concerning the clinical application of neural stem cells (NSCs) to the brain, spinal cord and peripheral nerves, and its application to internal ear also has been paid great attention. It is feasible to utilize NSCs transplantation for repair of damages to internal ear hair cells, spiral ganglion and auditory nerve. OBJECTIVE: To review characteristics, culture of NSCs and its application to damage to auditory nerve in the internal ear. METHODS: The first author retrieved PubMed Database (http://www.ncbi.nlm.nih.gov/PubMed) and Wanfang Database (http://www.wanfangdata.com.cn) published from 1990 to 2008 for articles addressing culture and differentiation of NSCs, and its application in the internal ear. The key words were “neural stem cells, transplant, inner ear, peripheral nerve”. Repetitive studies were excluded, and 107 articles were primarily collected. Following reading titles and abstracts, a total 36 articles were used for review. RESULTS AND CONCLUSION: Biological features of NSCs are understood, but the studies concerning NSCs have obtained great progress. Stem cell transplantation can be used in the study of auditory sensation. Regeneration or protection of hair cells and connected spiral neurons provide extensive prospects for treating internal ear disease. However, it is a challenge to realize hair cell regeneration and reconstruction of auditory pathway. With the renewal and development of NSCs basic theory, NSCs application to clinic would obtain breakthrough. Related Articles | Metrics

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Colonization factors of stem cells in myocardial infarction tissues He Ping, Li He, Zhao Yu-sheng 2010, 14 (27):  5090-5094.  doi: 10.3969/j.issn.1673-8225.2010.27.035 Abstract ( 95 )   PDF (638KB) ( 354 )   Save BACKGROUND: Pathological changes, such as myocardial lesions or absence, are commonly in heart disease. The myocardial cells are terminal cells that can not regenerate. Thus, it is a hot focus to explore an effective mechanism for myocardial tissue regeneration and repair. OBJECTIVE: To review the progress of myocardial tissue-engineered materials and the promotive effects of cytokine and Chinese patent medicine on stem cells homing, and to provide a theoretical basis for the study of stem cells differentiate into myocardial cells. METHODS: Documents in the PubMed and VIP databases from January 2002 to July 2009 were searched by computer using key words of “Stem Cells, Myocardial infarction, Tissue Engineering and Cytokines” both in English and Chinese. Papers related to myocardial regeneration, scaffold materials, revascularization of myocardial tissue, and the effects of cytokine and Chinese patent medicine on stem cells differentiation were included. Repetitive research or Meta analysis was excluded. RESULTS AND CONCLUSION: Stem cell transplantation has shown attractive prospect in the treatment of myocardial infarction. However, generally, stem cells could seldom enter the peripheral circulation, and it was very limited to repair myocardial necrotic tissue. There were three key issues: induction and differentiation, transplantation and survival, mobilization and homing. Tissue engineering, cytokine, Chinese patent medicine, combined with stem cells transplantation, could be the effective treatment for myocardial infarction. Such problems unsolvable by far would be tackled by mobilization and homing of stem cells. But the research of stem cells just begin now, many puzzles need to be solved. Related Articles | Metrics

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Immunological characteristics of adipose-derived stem cells Han Chang-jie, Yang Xiang-qun, Zhang Chuan-sen 2010, 14 (27):  5095-5098.  doi: 10.3969/j.issn.1673-8225.2010.27.036 Abstract ( 116 )   PDF (545KB) ( 601 )   Save BACKGROUND: Due to the immunological rejection caused by allogeneic cells, both of stem cell transplantation and tissue engineering are limited to the scope of individualized treatment and therefore not suitable for treatment of war trauma or extensive clinical use. Adipose-derived stem cells not only have the multiple differentiation potential, but also characterized by their broad sources, enough amount, easy-to-amplification and low immunogenicity. Thus, it is possible to construct heterologous tissue-engineered organs. OBJECTIVE: To summarize progresses in the immunological characteristics of adipose-derived stem cells. METHODS: A computer-based online search of VIP, Pubmed and Elsevier databases was performed for related articles with the key words of “adipose-derived stem cells, ADSCs, immunogenicity, immune regulation, implantation”. Articles related to immunological characteristics of adipose-derived stem cells were included, and repetitive studies were excluded. RESULTS AND CONCLUSION: A total of 26 articles were collected, and 17 were included. At present, the immunological characteristics researches of adipose-derived stem cells mainly focus on the immunophenotype, in vitro and in vivo immunological properties, revealing that adipose-derived stem cells have low immunogenicity and immune regulation property, hopefully to be a candidate for allogeneic stem cell transplantation and tissue engineering products. Related Articles | Metrics

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Stem cells and nerve-related factors effects on exercise-induced peripheral nerve injury regeneration He Chuan-sheng, Liu Rui-lian 2010, 14 (27):  5099-5102.  doi: 10.3969/j.issn.1673-8225.2010.27.037 Abstract ( 61 )   PDF (585KB) ( 433 )   Save BACKGROUND: Current stem cell technology in heart and brain diseases, central nervous system damage and peripheral nerve injury plays an important role. However, the exercise-induced peripheral nerve injury research is still in the basic stage. OBJECTIVE: To study the current stem cell technologies used in exercise-induced regeneration of peripheral nerve injury, in order to lay the scientific basis of rehabilitation of exercise-induced peripheral nerve injury. METHODS: We searched PubMed database by computer for related literatures published from January 1991 to January 2010. The literature content was closely related to the study of stem cells and exercise-induced peripheral nerve injury. A total of 26 documents were selected. RESULTS AND CONCLUSION: Diseases of peripheral nerve injury often occur during exercise training, while stem cell transplantation and induction differentiation technologies can facilitate the regeneration of peripheral nerve injury, and plays an important role in the promotion of sports training in the prevention and treatment of peripheral nerve injury. However, it is still in the basic research stage, and requires further studies. Related Articles | Metrics

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Effects of endothelial progenitor cells in treatment of vascular-related diseases Lu Jing-Jing, Yang Zhen-dong, Wang Gui-min 2010, 14 (27):  5103-5106.  doi: 10.3969/j.issn.1673-8225.2010.27.038 Abstract ( 88 )   PDF (607KB) ( 489 )   Save BACKGROUND: Endothelial progenitor cells have been extensively applied to the treatment of vascular-related diseases in recent years, due to its high proliferation potential and multidirectional differentiation. OBJECTIVE: To summarize the research progress of endothelial progenitor cells in treatment of vascular-related diseases. METHODS: We retrieved PubMed Database for relevant articles published from January 2004 to September 2009. The key words were “endothelial progenitor cells, vascular-related disease” in English. Simultaneously, a computer-based search was performed in Tsinghua Tongfang Full Text Database for relevant articles published from January 2004 to September 2009. The key words were “endothelial progenitor cells, vascular-related disease” in Chinese. The literatures were classified according to inclusion and exclusion criteria by the first and second authors. Its contents and results were evaluated. Finally, a total of 30 articles were included. RESULTS AND CONCLUSION: Endothelial progenitor cells are a vascular endothelial precursor cells. It has been confirmed that endothelial progenitor cells are not only involved in vasculogenesis in embryonic period, but also participate in angiogenesis after birth. It is in a dynamic equilibrium state between endothelial damage and the repair effects of surrounding mature endothelial cells and endothelial progenitor cells to vascular endothelial in normal circumstances. In pathological conditions, the function of proliferation and migration, adhesion of endothelial progenitor cells are all weaker than before, leading to that vascular endothelial injury cannot be repaired or damaged, thus become an important dangerous factor in many vascular-related diseases. Endothelial progenitor cells have great application value and prospect in treatment of vascular-related diseases and deserve further investigations. Related Articles | Metrics

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Gene expression on transcriptional level regulated by noncoding small RNA in the proliferation and differentiation of cells and stem cells  Wang Lian-dong, Zhang Ming-yuan 2010, 14 (27):  5107-5110.  doi: 10.3969/j.issn.1673-8225.2010.27.039 Abstract ( 68 )   PDF (567KB) ( 418 )   Save BACKGROUND: Up to now, numerous studies have addressed small RNA-mediated gene silencing following transcription, but few reports have concerned regulation of noncoding small RNA on other gene expression levels. OBJECTIVE: To review gene regulation of noncoding small RNA on transcriptional level and its effects on proliferation and differentiation of stem cells. METHODS: PubMed Database was undertaken to identify relevant articles on RNA interference mechanism and transcriptional gene silencing mechanism published from January 1998 to December 2009. The key words were “RNAi, transcriptional gene silencing” in English. The data were primarily examined, and references of each article were looked through. Repetitive researches were excluded, and 69 articles were relevant ones. Thus, a total of 28 articles were in accordance with inclusion criteria. RESULTS AND CONCLUSION: Many species have transcription gene silencing, and the transcription gene silencing induced by small interference RNA in mammalian cells is a universal phenomenon. Furthermore, Dicer family and Agronaute family are two indispensable gene families to result in transcription gene silence, which also regulate the generation and differentiation of stem cells. Noncoding small RNA induces a high genetic transcriptional activity, and the regulation of small RNA has two modes, RNA/DNA mode and RNA/RNA mode. Related Articles | Metrics

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Haploidentical peripheral stem cell transplantation for treatment of hematologic malignancies in six cases Wang Zhi-ming, Wang Lin, Chen Xiao-xia, Luo Xian-sheng, He Li-li, Xu Dan-dan, Li Xing, Fu Rong-xiang, Wang Yun-ying, Li Li-qiong, Huang Zi-ying, Tan Lian 2010, 14 (27):  5111-5114.  doi: 10.3969/j.issn.1673-8225.2010.27.040 Abstract ( 63 )   PDF (432KB) ( 396 )   Save BACKGROUND: Haploidentical hematopoietic stem cell transplantation is confronted with many problems such as difficulty to implant, slow reconstitution, severe graft versus host disease, delayed immunologic reconstitution, and high incidence rate of lethal infection. To overcome these problems can widely use the transplantation of haploidentical hematopoietic stem cells. OBJECTIVE: To study the effect of haploidentical hematopoietic stem cell transplantation in treatment of hematologic malignancies. METHODS: We used cytosine arabinoside, busulphan, cyclophosphamide，the anti-thymocyte globulin and methyl-n-(2-chloroethlyl)-n-cyclohexyl-n-nitrosourea as preconditioning of patients, used cyclosporine A, mycophenolate mofetil, the anti-thymocyte globulin, interleukin-11 and methotrexate as prophylaxis of acute graft versus host diseases to treat 6 cases of hematologic malignancies. RESULTS AND CONCLUSION: All patients achieved complete engraftment. The median times of neutrophil recovery > 1.0×109/L were 15.8 (+12-+20) days after transplantation. The incidence of grade Ⅲ-Ⅳgraft-versus-host disease was 16.7%. All patients survived disease-free with a median follow-up of 34.7(13-63) months. Results have indicated that haploidentical hematopoietic stem cell transplantation is a safe and effective treatment for hematologic malignancies, with cytosine arabinoside, busulphan, cyclophosphamide, the anti-thymocyte globulin, methyl-n-(2-chloroethlyl)-n-cyclohexyl-n-nitrosourea as preconditioning, with cyclosporine A, mycophenolate mofetil, the anti-thymocyte globulin, interleukin-11 and methotrexate as prophylaxis of acute graft versus host diseases. Related Articles | Metrics

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In vitro isolation of intestinal side population cells from newborn mice Zhang Yong-feng, Yang Jin, Wu Zheng-zhi, Jia Xiu-qin, Li Ming, Li Ying-hong 2010, 14 (27):  5115-5118.  doi: 10.3969/j.issn.1673-8225.2010.27.041 Abstract ( 131 )   PDF (395KB) ( 358 )   Save BACKGROUND: Current methods of stem cell separation are mainly based on their cell markers. A method for stem cells separation which is not based on cell markers developed in recent years, that is fluorescence activated cell sorting method, has been applied for stem cells and mature cells separation. OBJECTIVE: To isolate side population cells from newborn mice small intestinal mucosa, and to investigate the feasibility of constructing the murine intestinal stem cell population by fluorescence activated cell sorting. METHODS: Small intestine mucosa organoids of mice were isolated and dissociated into single cells. The side population cells were stained with Hoechst 33342 and propidium iodide, then sorted using fluorescence activated cell sorting. Total RNA and protein were purified from sorted fractions to detect Musashi-1 expressions by RT-PCR and Western-blotting. RESULTS AND CONCLUSION: Single cell suspension from mouse small intestine mucosa contained a viable population of cells, which showed the side population phenotype and were sensitive to verapamil. These cells were enriched for Musashi-1 mRNA and MSI-1 protein expression. Results demonstrated that the side population fraction separated from mice intestinal mucosa is enriched for intestinal stem cells, the murine intestinal stem cell population can be successfully constructed with fluorescence activated cell sorting. Related Articles | Metrics

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Clinical verification of olfactory ensheathing cell transplantation in treatment of spinal cord injury Zheng Zun-cheng1, Wei Kai-bin, Liu Feng, Liu Chao, Wei Shu-gang, Cheng Zong, Gao Rui, Zhang Lei, Zhang Kun, Kuang Nai-feng, Zhang Li-qing 2010, 14 (27):  5119-5122.  doi: 10.3969/j.issn.1673-8225.2010.27.042 Abstract ( 72 )   PDF (279KB) ( 502 )   Save BACKGROUND: A series of basic researches have confirmed that, the olfactory ensheathing cell transplantation can promote spinal cord regeneration and recover some neurological functions of spinal cord in animal models of spinal cord injury. Some clinical trials also prove that transplantation of olfactory ensheathing cells can indeed improve neurological function in patients with spinal cord injury, and then improve their quality of life. OBJECTIVE: To verify the effectiveness and safety of olfactory ensheathing cell transplantation in repair of neurological function of spinal cord injury patients. METHODS: The aborted embryonic olfactory bulb was collected and digested into single olfactory ensheathing cells. After they were cultured and purified 2 weeks, olfactory ensheathing cell suspension was prepared. A total of 213 cases of spinal cord injury were selected. Under general anesthesia, the prepared olfactory ensheathing cell suspension was injected through several target sites surrounding the injured spinal cord. ASIA scale was used to assay the patients before transplantation, 3 weeks to 2 months after transplantation, so as to evaluate spinal cord recovery. RESULTS AND CONCLUSION: The spinal cord nerve function in all patients altered to different degrees at 3 weeks postoperation. Spinal cord function score, the sensory and motor functions were significantly increased compared with preoperation (P < 0.001), and showed a trend of continuous improvement with time; the patients were visited as follow-up for no more than 5 years, and no impairment of the restored nervous function or transplant adverse reactions were observed. It is confirmed that olfactory ensheathing cell transplantation can promote the recovery of nerve function in patients with spinal cord injury, it can restore and improve some spinal cord functions, and the treatment is safe. Related Articles | Metrics

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Pulmonary Aspergillus infection coexisted with obliterative bronchiolitis in a patient at one year following allogeneic hematopoietic stem cells transplantation Xiao Hao-wen, Jiang Zu-jun, Xiao Yang, Gao Yang, Zhang Xiao-ming, Pang Yan, Ouyang Ling 2010, 14 (27):  5123-5126.  doi: 10.3969/j.issn.1673-8225. 2010.27.043 Abstract ( 104 )   PDF (330KB) ( 457 )   Save BACKGROUND: Allogeneic hematopoietic stem cell transplantation (allo-HSCT) is an effective method for treating multiple malignant hematological diseases and hereditary diseases. However, systematic internal organs disorders, especially pulmonary complications, are commonly following allo-HSCT. How to correctly diagnose and treat the coexistence of pulmonary infectious complications and pulmonary noninfectious complications has great importance.  OBJECTIVE: To report a case suffered from pulmonary Aspergillus infection coexisted with obliterative bronchiolitis at 1 year following allo-HSCT, and to discuss the prevention, clinical manifestation and treating method by reviewing related literature. METHODS: At 373 days after allo-HSCT, the patient developed fever, dry cough, shortness of breath and dyspnea on exertion. A high-resolution computed tomography of chest demonstrated that there were alveolar infiltrating in the upper, middle and lower lobe of the right lung, and the focus of infection was performed further biopsy. RESULTS AND CONCLUSION: The histopathological examination of the sample showed alveolus dilatation, epithelial cells hyperplasia, fibrinous obliteration in alveolar space and peribronchiolar lymphocytes inflammation, which were CD3(+), CD45RO(+), CD20(-), CD79a(-), MPO(-), CD34(-). Aspergillus fumigatus could be seen in the cultured biopsied tissue specimen. Pulmonary function test showed that, air flow obstruction with reduction of forced expiratory volume in one second was 59.27%. The patient was diagnosed as invasive pulmonary Aspergillus infection combined with bronchiolitis obliterans and was treated by caspofungin combined with intravenous voriconazole for invasive aspergillosis, methylprednisolone, azathioprine, intravenous immunoglobulin and azithromycin for bronchiolitis obliterans. At 40 days after treatment, the CT examination showed the focus was absorbed completely. Related Articles | Metrics

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Protective effect of oyster extract on apoptosis of cerebral neural stem cells induced by hyperthermia Song Hai-yan, Song Yong-li, Yu Qing-mei, Zhuang Yuan, Wu Yu-ling 2010, 14 (27):  5127-5130.  doi: 10.3969/j.issn.1673-8225. 2010.27.044 Abstract ( 102 )   PDF (271KB) ( 358 )   Save BACKGROUND: Previous results of our study show that oyster extract has some protective effects on apoptosis of the neuroepithelium in neural tube defects induced by hyperthermia in vivo. It remains unclear whether the extract also protects in vitro cultured neural stem cells. OBJECTIVE: To investigate the protective effect of oyster extract on apoptosis of cerebral neural stem cells induced by hyperthermia. METHODS: The cerebral neural stem cells of embryonic mice of 13 days were cultured in vitro. Nestin expression was detected by immunofluorescence method to identify neural stem cells. The neural stem cells of passage 3 were divided randomly into 4 groups: hyperthermia control group and oyster treated I, II, III groups (mass concentration 2.5, 5, 10 g/L oyster extract solution). In addition, culture solution control group (no cells), and culture solution+ oyster extract control group (no cells) were designed. All oyster extract groups and control groups were treated by hyperthermia over 39 °C. The survival rate and the vitality of neural stem cells were detected by trypan blue staining and MTT assay. Western-blotting was employed to explore the expression of p53 in cerebral neural stem cells of each group. RESULTS AND CONCLUSION: The survival rate and the value of MTT assay in oyster treated groups II and III were significantly greater than hyperthermia control group (P < 0.05), but the expression of p53 in oyster treated groups II and III were weaker than hyperthermia control group. Oyster extract plays an important protective role in the apoptosis of neural stem cells induced by hyperthermia. Related Articles | Metrics