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07 May 2011, Volume 15 Issue 19 Previous Issue    Next Issue

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A comparative study on bone marrow mesenchymal stem cells differentiated into chondrocytes induced by co-culture with chondrocytes and conditioned medium Lü jing, Ke Jie, Wang Lin, Zhang Dong-mei, Liu Bing, Pang Jian-liang, Deng Tian-zheng 2011, 15 (19):  3421-3426.  doi: 10.3969/j.issn.1673-8225.2011.19.001 Abstract ( 83 )   PDF (1323KB) ( 463 )   Save BACKGROUND: In vitro differentiation of bone marrow mesenchymal stem cells (BMSCs) is dependent on appropriate culture conditions to a great extent. OBJECTIVE: To study the biological characteristics of chondrocytes differentiated from BMSCs co-cultured with chondrocytes versus cultured in conditioned medium. METHODS: Rat BMSCs and chondrocytes were isolated and cultured in neural induction. Cellular non-contact co-culture induced method and conditioned medium induced method were taken to reduce BMSCs differentiating into chondrocytes. And the biological characteristics of the cells were evaluated by MTT, mitotic cycle, GAG secretion, RT-PCR, cell-staining methods, immunocytochemistry. RESULTS AND CONCLUSION: The biological characters of co-culture induced cells were seemed to be better than those of conditioned medium induced cells, which point out that BMSCs co-cultured with chondrocytes are appropriate as seed cells in cartilage tissue engineering. Related Articles | Metrics

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In vitro differentiation of fetal bone marrow mesenchymal stem cells into islet beta like cells Guo Jia-ning, Lei Jian-yuan, Li Yi, Chang Ji-wu 2011, 15 (19):  3427-3432.  doi: 10.3969/j.issn.1673-8225.2011.19.002 Abstract ( 76 )   PDF (2084KB) ( 335 )   Save BACKGROUND: There is no a mature program for identifying islet like cells differentiated from human bone marrow mesenchymal stem cells (BMSCs). OBJECTIVE: To investigate the possibility of human fetal BMSCs differentiating into insulin-secreting cells in appropriate condition. METHODS: The fetal BMSCs were co-cultured with fetal insulin cells on Transwell double-layer culture plate. In the control group, the BMSCs were cultured in culture medium containing islet cells. Morphological changes in fetal BMSCs and islet cells were analyzed under a phase contrast microscopy. The morphological changes and insulin secretions of BMSCs under the stimulation of fetal islet cells were detected by radioimmunoassay. RESULTS AND CONCLUSION: The fetal BMSCs were isolated and purified, identical cell morphology liked plants spindle-like arrangement. Fetal islet cells were confirmed by panccreatic β endocrine cells under electron microscopy. The undifferentiated BMSCs exhibited were adherent and long spindle-shaped cells. After induction, cells gradually became round and formed clusters. Cells secreted abundant insulin and formed islet-like cell clusters that exhibited positive dithizone staining and immunocytochemical staining at 9 days. No insulin was detected in the control group. It is indicated that fetal BMSCs have the potentiality to differentiate into insulin-producing cells in appropriate condition． Related Articles | Metrics

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Construction of human LIM mineralization protein-1 gene recombinant adenovirus and its expression in dog bone marrow mesenchymal stem cells Pu Chao, Ni Wei-dong, Gao Shi-chang, Qiu Yu 2011, 15 (19):  3433-3437.  doi: 10.3969/j.issn.1673-8225.2011.19.003 Abstract ( 98 )   PDF (1492KB) ( 363 )   Save BACKGROUND: Studies shows that the LIM mineralization protein 1 (LMP-1) can stimulate bone formation in vivo and in vitro. OBJECTIVE: To construct the recombinant adenovirus vector containing LMP-1 gene by using the Ad-Easy system, then to detect the gene expression and study the mechanism of bone formation induced by LMP-1 in infected dog bone marrow mesenchymal stem cells (BMSCs). METHODS: The recombinant plasmid pAd-LMP-1 was constructed. The replication-defective recombinant adenovirus Ad-LMP-1 was packaged and amplified in the human embryonic kidney 293 cells (HEK293). Dog BMSCs were infected by Ad-LMP-1 in the best value of MOI in vitro. The gene expression of LMP-1and BMP-7 was detected by RT-PCR and Western Blot. Dogs BMSCs were treated with Ad-LMP-1 and recombinant human bone morphogenetic protein 7 (rhBMP-7) to study the mechanism of bone formation of LMP-1 undergoing mineralization (Ca) nodules at 21 days by alizarin red staining. RESULTS AND CONCLUSION: ①Ad-LMP-1 was successfully constructed and obtained titers in the packaging about 1×1011 efu/mL after amplified in HEK293. ②The best efficiency could be obtained at the most appropriate MOI 100 after Ad-LMP-1 infected BMSCs. Infected BMSCs can express mRNA and protein of LMP-1, but not caused expression of BMP-7. ③ Dog BMSCs were treated with Ad-LMP-1 and (or) rhBMP-7 for 21 days. We found that Ad-LMP-1 with exogenous rhBMP-7 promote transformation into osteoblasts together. Neither Ad-LMP-1 nor rhBMP-7 alone induced any transformation on day 21. The recombinant adenoviral vector containing hLMP-1 gene was successfully constructed and expressed effectively in dogs BMSCs. It is confirmed that the mechanism of bone formation induced by LMP-1 is synergistic effect with the performance of exogenous rhBMP-7. Related Articles | Metrics

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Growth characteristics of bone marrow stromal stem cells in neonatal rats with hypoxic-ischemic encephalopathy Zhang Yi, Yao Li 2011, 15 (19):  3438-3442.  doi: 10.3969/j.issn.1673-8225.2011.19.004 Abstract ( 95 )   PDF (1827KB) ( 303 )   Save BACKGROUND: The growth features of bone marrow stromal stem cells (BMSCs) for hypoxia ischemic encephalopathy is unclear. OBJECTIVE: To observe growth characteristics and physical and chemical characters of BMSCs in treatment of acute anoxic-ischemia newborn rats. METHODS: Seven-day-old rats were selected to ligate the bilateral carotid arteries to prepare ischemic models. After 2 hours, the neonatal rats deeply suffered from hypoxia (8% O2 and 92% N2) for 1-2.5 hours. The rats were divided into mild group (ischemia for 1 hour), moderate group (ischemia for 1.5 hours), severe group (ischemia for 2.5 hours), anoxia-ischemia group and sham operation group (only ligation of the left carotid artery). The growth condition of BMSCs was observed. RESULTS: After 3-hour culture, BMSCs grew adherently in the sham operation and anoxia-ischemia groups, superior to those in the moderate and severe groups. The morphology of BMSCs recovered in the sham operation, mild and moderate groups. The apoptotic rates were similar in the sham operation and mild groups, and obviously increased in the moderate and severe groups. The activity of supernatant lactate dehydrogenase was increased in the mild and severe groups to different extent. The findings demonstrate that the effect of mild and moderate anoxia-ischemia on BMSCs is reversible, and BMSCs grow vigorously and benefit to injury repair. Related Articles | Metrics

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Bone marrow-derived stem cells involved in the renewal of vascular endothelium Xu Yi-ling, He Xiang-hui, Wei Wei, Zhang Zhi-xiang, Zhu Li-wei 2011, 15 (19):  3443-3446.  doi: 10.3969/j.issn.1673-8225.2011.19.005 Abstract ( 80 )   PDF (1145KB) ( 284 )   Save BACKGROUND: Bone marrow-derived stem cells have strong plasticity, but the mechanism that they participate in regeneration and renewal of tissue is still conflicting between transdifferentiation and cell fusion. OBJECTIVE: To investigate the possibility and possible mechanism of bone marrow-derived stem cells involvement in the renewal of vascular endothelial cells in different organs through sex crossing bone marrow transplantation using mouse models of sexual mosaic. METHODS: Female C57BL/6-green fluorescent protein (GFP) mice were used as donor mice, and male C57BL/6 mice were used as the recipient to establish bone marrow transplanted chimeric mice. The chimerism of bone marrow was assessed by flow cytometry. Twenty weeks after transplantation, frozen slide of the brain, kidney, liver, spleen, and heart were prepared and fluorescence in situ hybridization was applied to label the Y chromosome, then GFP expression and Y chromosome were observed using fluorescence microscope. RESULTS AND CONCLUSION: The percent of GFP+ cell of bone marrow cells in the recipient was (7.48±1.38)% and (73.92±5.57)% at week 1 and week 4 respectively, indicating the success of sex crossing bone marrow transplantation model. In week 20, GFP expression was detected in vascular endothelial cells in the brain, kidney, liver, and spleen, and some cells also showed Y chromosome expression, indicating cell fusion occurred. No GFP expression was observed within the parenchyma of the brain and heart. Results have indicated that bone marrow-derived stem cells might have participated in the renewal of vascular endothelial cells in different organs through cell fusion. Related Articles | Metrics

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A preliminary study of the number and identification of mesenchymal stem cells in spinal surgery bleeding Ye Bin, Qin Jian-pu, Cai Yu-qiang, Zhang Jun, Chen Lin, Liao Wen-bo 2011, 15 (19):  3447-3450.  doi: 10.3969/j.issn.1673-8225.2011.19.006 Abstract ( 65 )   PDF (983KB) ( 299 )   Save BACKGROUND: A great amount of red bone marrow in the cancellous bone losses during orthopedic surgery. OBJECTIVE: To explore the feasibility to isolate bone marrow mesenchymal stem cells (BMSCs) during spinal surgery bleeding and to investigate the relationship of the number of BMMSCs with age and time. METHODS: Three cases of degenerative lumbar instability and 16 cases of lumbar burst fractures were selected to undergo posterior pedicle screw fixation, including 4 cases of 20-30 years, 5 of 30-40 years, 5 of 40-50 years, 2 of 50-60 years, 1 of 64 years, 1 of 65 years, and 1 of 67 years. 5 mL of vertebra blood was collected from 4 cases within 0-5 minutes after the awl for pierced vertebra was pulled out, and 5 mL of vertebral blood from other 15 cases was collected within 0-5, 5-10 and 10-15 minutes after the awl was pulled out, respectively. Mononuclear cells were isolated and incubated into 6-well plates, and then cultured for 3 days. The medium was renewed completely to remove unadherent cells. Hematoxylin-eosin staining was applied for cell counting. RESULTS AND CONCLUSION: The membrane mark showed that CD44 and CD29 were positive while the CD34 and CD14 were negative. The number of BMSCs isolated within 0-5 minutes was 262.47±172.20, and fewer or no BMSCs were found within 5-10 or 10-15 minutes. The number of BMMSCs from vertebra bleeding of 0-5 minutes had a decreased tendency with aging. Related Articles | Metrics

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Real-time PCR detection of GATA-1, GATA-2, peroxisome proliferator-activated receptor gamma, SCF gene in bone marrow mesenchymal stem cells of aplastic anemia patients and normal people He Li, Xiao Yang, Jiang Zu-jun, Li Li, Li Yong-hua, Ouyang Ling, Gao Yang, Kuang Li-ping 2011, 15 (19):  3451-3454.  doi: 10.3969/j.issn.1673-8225.2011.19.007 Abstract ( 70 )   PDF (1243KB) ( 316 )   Save BACKGROUND: GATA-1, GATA-2, peroxisome proliferator-activated receptor gamma (PPAR-γ), and SCF gene play an important role in bone marrow hematopoietic and concomitant regulation mechanism. OBJECTIVE: To observe the expressions of GATA-1, GATA-2, PPAR-γ, SCF gene in bone marrow mesenchymal stem cells of aplastic anemia patients and normal people. METHODS: we collected bone marrow samples from six aplastic anemia patients and six normal persons. Ficoll stick wall method was used to isolate and culture bone marrow mononuclear cells that began to passage at 70%-80% confluence. The 3rd generation of mesenchymal stem cells were detected using flow cytometry. Total RNA was extracted to synthesis and design primer sequence, analysis standard curve, amplification curve, dissolved curve, to be compared with NaCan each gene genetic differences. Fluorescent quantitative PCR detection was employed for GATA-1, GATA-2, PPAR-γ, SCF gene in bone marrow mesenchymal stem cells of aplastic anemia patients and normal people. RESULTS AND CONCLUSION: Expression of GATA-2 and SCF gene was lower (P=0.012, 0.039) and PPAR-γ expression was higher (P=0.035) in the aplastic anemia patients as compared with normal persons. There was no difference in GATA-1 expression. The findings indicated that abnormal expression of GATA-2 and SCF gene could affect macro-control of bone marrow hematopoietic microenvironment, and abnormal expression of PPAR-γ could explain why bone marrow adipogenesis easily formed in aplastic anemia patients. Related Articles | Metrics

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Superparamagnetic iron oxide and 4',6-diamidino-2-phenylindole double-labeled bone marrow derived mesenchymal stem cells Yang Rong, Li Jian-ding, Zhang Rui-ping 2011, 15 (19):  3455-3458.  doi: 10.3969/j.issn.1673-8225.2011.19.008 Abstract ( 90 )   PDF (1248KB) ( 517 )   Save BACKGROUND: Now, bone marrow mesenchymal stem cells (BMSCs) have become important seed cells of tissue engineering. It is necessary to solve the problem that how to label BMSCs efficiently and safely for further study on BMSCs proliferation and differentiation in vitro and in vivo. OBJECTIVE: To investigate the effects of superparamagnetic iron oxide (SPIO) and 4',6-diamidino-2-phenylindole (DAPI) double-labeling on BMSCs, and to study whether there is any effect on cell viability, proliferation and apoptosis. METHODS: BMSCs were isolated and double-labeled with SPIO and DAPI. The labeling rate was identified by Prussian blue staining and fluorescence microscope, then cell vitality was detected by Trypan blue staining and proliferation activity was measured by MTT assay. Cell survival rate and apoptosis rate were observed by using Calceein AM/PI and AO/PI staining. RESULTS AND CONCLUSION: The stem cell double-labeling rate by SPIO and DAPI were nearly 100%. The cell survival rate with Trypan blue staining was 97%. MTT method detected that there was no significant difference for proliferation activity between double-labeled and unlabeled stem cells (P > 0.05). Calcein-AM/PI staining detected that the survival rate of double-labeled and unlabeled stem cells were 95% and 96%. AO/PI staining detected that the apoptosis rate of double-labeled and unlabeled stem cells were both 1%. These suggested that there is no effect on survival, proliferation and apoptosis of BMSCs after SPIO and DAPI double-labeling. Related Articles | Metrics

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Construction and identification of eukaryotic expression vector of RNA interference specific for estrogen receptor beta Zheng Hong, Li Xiu-hua, Yin Li-hua, Li Jing, Ding Yin 2011, 15 (19):  3459-3462.  doi: 10.3969/j.issn.1673-8225.2011.19.009 Abstract ( 132 )   PDF (1067KB) ( 283 )   Save BACKGROUND: Whether estrogen receptor beta (ER-β) participates to mediate proliferation and differentiation of bone marrow mesenchymal stem cells is unclear. OBJECTIVE: To construct and identify a specific small interfering RNA (siRNA) suppressing estrogen receptor beta gene. METHODS: Genome sequences of ER-β gene was retrieved from Genbank and cDNA was designed coding expression of small hairpin RNAs (shRNA) for ER-β gene. It was cloned into the expression vector pSilencer 3.1-H1 into a short hairpin RNA with BamH Ⅰ, HindⅢ sticky end, terminated cognition sequence and a loop structure, followed by being cloned in pRNAT-U6.1 /Neo to construct a expression vector of ER-β-specific siRNA. At last the vector was evaluated by enzyme cutting and gene sequencing test. RESULTS AND CONCLUSION: The recombinant plasmid of RNA interference specific for ER-β identified by the restriction map completely coincided with the designs. The siRNA eukaryotic expression vector against ER-β mRNA has been successfully constructed. Related Articles | Metrics

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Retroviral-mediated tissue inhibitor of metalloproteinase-1 gene transfected human mandible bone marrow mesenchymal stem cells and their osteogenic potential Gao Wen-ling, Liu Fa-keng, Hu Kai-shun, Zhu Shuang-lin 2011, 15 (19):  3463-3466.  doi: 10.3969/j.issn.1673-8225.2011.19.010 Abstract ( 155 )   PDF (1241KB) ( 552 )   Save BACKGROUND: Extracellular matrix (ECM) accumulation of mesenchymal stem cells (MSCs) occurs at the early phase of osteogenesis. And the main components of ECM deposited in that process are specific substrates of matrix metalloproteinases (MMPs) suppressed by tissue inhibitor of matrix metalloproteinase-1 (TIMP-1). Whether MSCs can inhibit the activity of MMPs and improve the ability of osteogenic differentiation by increasing the expression of TIMP-1? OBJECTIVE: To investigate the osteogenic potential of human mandible bone marrow mesenchymal stem cells (hMBM-MSCs) modified by TIMP-1 gene mediated with retroviral transfection. METHODS: In-Fusion primers for TIMP-1 gene was designed, and reacted with digested linearized pBABE-Puro retroviral expression vector by homologous recombination sites and verified TIMP-1 expression with reverse transcription-polymerase chain reaction (RT-PCR) and sequencing methods, and then packaged with GP293 cell line. Bone marrow mesenchymal stem cells in mandibular surgery patients were separated by density gradient method, and their surface markers were verified by flow cytometry. TIMP-1 packaging liquid after retrovirus transfection infected and selected hMBM-MSCs cell lines which stably expressed TIMP-1 gene, and RT-PCR and protein electrophoresis validate its expression. HMBM-MSCs were induced using osteogenic induction medium for 10 days, and their osteogenic potential was observed. The expression of osteogenic related genes was verified by RT-PCR. RESULTS AND CONCLUSION: Confirmed by DNA sequencing, RT-PCR obtained cDNA sequences of TIMP-1 gene totally matched with NM_003254.2 (624 bp) identical sequences. HMBM-MSCs were successfully separated according to surface makers identification, and manifested a good osteogenic and adipogenic ability. RT -PCR and Western Blot were used to verify the successful expression of hMBM-MSCs by retroviral vector-mediated transfection. RT-PCR showed that when hMBM-MSCs were induced with osteogenic induction media for 10 days, TIMP-1 increased markedly the mRNA expression of RUNX-2 and COL1. TIMP-1 gene can be successfully transfected into hMBM-MSCs by retroviral transfection, and promote the effect of osteogenesis. Related Articles | Metrics

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Expression of nm23-H1 during differentiation of human umbilical cord mesenchymal stem cells into adipocytes and osteoblasts in vitro Rong Jing, Peng Xin-lei, Ma Yan-yan, Liu Qiu-ying, Zhou Xiang-rong, Luo Xin, Wang Yi-fei 2011, 15 (19):  3467-3471.  doi: 10.3969/j.issn.1673-8225.2011.19.011 Abstract ( 144 )   PDF (1688KB) ( 573 )   Save BACKGROUND: It has been proved that nm23-H1 gene has a great infulence on proliferation, differentiation and transfering of many kinds of cells. OBJECTIVE: To study on the expression of nm23-H1 gene during differentiation of human umbilical cord mesenchymal stem cells (hUCMSCs) into adipocytes and osteoblasts in vitro. METHODS: The hUCMSCs were isolated from human umbilical cord by tissue adherence, and a flow cytometry was used for immunophenotype identification. Then the 3rd passage cells were induced to differentiate into adipocytes and osteoblasts, characterized by oil red O staining and Alizarin red staining respectively. The expression of nm23-H1 mRNA was detected by real time quantitative RT-PCR during the adipogenic and osteogenic differentiation. RESULTS AND CONCLUSION: During the adipogenic differentiation, the nm23-H1 expression was increased in mRNA level after the induction on 4 days, and reached to the highest level on 7 days (P < 0.01), and then was gradually decreased on 14 days to the primary level which had no significant changes compared with untreated control (P > 0.05). During the osteogenic differentiation, the nm23-H1 expression in mRNA level was downregulated (P < 0.01) until maturation on 28 days (P > 0.05). nm23-H1 gene has shown a regulatory changes during the differerntiation of hUCMSCs into adipocytes and osteoblasts, this might imply that nm23-H1 gene make a contribution to the osteogenic and adipogenic differentiation. Related Articles | Metrics

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Comparison of two isolation methods for umbilical cord blood mesenchymal stem cells Gao Yang, Li Li, Ran Jiang-hua, Chen Yi-ming, Jiang Yi-zhou 2011, 15 (19):  3472-3475.  doi: 10.3969/j.issn.1673-8225.2011.19.012 Abstract ( 90 )   PDF (1207KB) ( 371 )   Save BACKGROUND: There is no standard method to isolate umbilical cord blood mesenchymal stem cells (MSCs)． OBJECTIVE: To discuss an optimized isolation method of human umbilical cord blood MSCs． METHODS: Monocytes were isolated from umbilical cord blood using the Percoll separation medium method and hydroxyethyl starch sedimentation respectively. Cells were cultured in DMEM/F12 containing 15% newborn calf serum. The main outcome indictors were recovery rate of monocytes, generation time, cell proliferation, morphological change, and the surface markers expressed on umbilical cord blood MSCs measured by flow cytometer. RESULTS AND CONCLUSION: Compared with the Percoll separation medium method, the recovery rate of monocytes increased and the first passage time reduced using the hydroxyethyl starch sedimentation (P < 0.01). But the morphological change and surface markers CD90, CD44, CD34 expression of two methods have no differences (P > 0.05). Therefore, the hydroxyethyl starch sedimentation has higher isolation efficiency and the shorter culture time than Percoll separation medium method. However, the quality of umbilical cord blood MSCs which are isolated by two methods has no difference. Related Articles | Metrics

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Prokaryotic expression and purification in Flt3 ligand extracellular domain and effects on cord blood CD34+ cells amplification Liu Qiang, Zheng Jin, Zhao Xing-cheng, Yin Dan-dan, Chen Ren-an, Liu Li, Hao Miao-wang, Liang Ying-min 2011, 15 (19):  3476-3480.  doi: 10.3969/j.issn.1673-8225.2011.19.013 Abstract ( 129 )   PDF (677KB) ( 368 )   Save BACKGROUND: Fms-like tyrosine kinase 3 ligand (Flt3 ligand, FL) is a cytokine that affects the growth, survival and/or differentiation of hematopoietic cells through the activation of specific tyrosine kinase receptors, and it is potentially useful for in vitro hematopoietic stem cells (HSCs) amplification. OBJECTIVE: To construct the recombinant prokaryotic expression vector pET32a(+)-hFLext, and to express and purify hFLext protein and investigate its effect on expansion of cord blood CD34+ cells. METHODS: The hFLext gene was cloned and the recombinant expression vector pET32a-hFLext was constructed and transformed into E. coli BL21. The hFLext protein was expressed under IPTG induction, purified by affinity chromatography with Ni beads. The cord blood CD34+ cells were isolated using CD34 microbead kit and incubated with hFLext alone or in combination with SCF and TPO. The effect of hFLext on expansion of CD34+ cells was examined after 1-week incubation. RESULTS AND CONCLUSIONS: The recombinant expression vector pET32a-hFLext was successfully constructed and the hFLext was successfully expressed as a Trx fusion protein (Trx-hFLext) in E. coli BL21. The Trx-hFLext with high purity was successfully obtained after degenerating proteins in inclusion bodies with 8M urea, renaturing by gradual dialysis, purifying by affinity chromatography with Ni beads. The purified hFLext can not only effectively maintain and slightly stimulate the proliferation of cord blood CD34+ cells, but also expand profoundly in combination with SCF and TPO, thus paving the way for its application in expanding human hematopoietic stem/progenitor cells in vitro further. Related Articles | Metrics

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Effects of mouse placenta-derived mesenchymal stem cells on the natural senescence of the mouse hematopoietic system Huang Hai-yan, Liu Cheng-cheng, Zhu Ai-zhen, Zhou Yan-hua, He Dong-mei, Liu Ge-xiu, Tan Guang-xiao 2011, 15 (19):  3481-3484.  doi: 10.3969/j.issn.1673-8225.2011.19.014 Abstract ( 127 )   PDF (645KB) ( 428 )   Save BACKGROUND: Hematopoietic system, as an important part of human body, will appear continuously functional recession with age. OBJECTIVE: To explore the anti-aging role of mouse placenta-derived mesenchymal stem cells (mPMSCs) on the natural senescence of the mouse hematopoietic system. METHODS: Experimental method: ①Preparation of the mPMSC: the placentas were collected from 6 pregnant 13.5 d Balb/c mice, and digested by 0.1% collagenase IV to obtain the cells, and the cells were plated, passaged, and identified as mPMSCs. ②Intervention of the mPMSCs: 6 month old Balb/c mice (48) were randomly divided into experimental and control groups. Mice of experimental group were injected with mPMSC (5×106 for each one) through tail intravenation, once a month, a total of six times; and mice of control group with physiological saline. Experimental assessment: ①The mPMSCs preparation: the morphology of cells was observed through inverted phase microscope, cell-surface markers by flow cytometry, cell differentiation potentials by inducing osteoblast differentiation and adipocyte differentiation. ②The mPMSCs intervention: on the third month after treatment, peripheral blood (WBC, Hb), bone marrow mononuclear cell count (BMNC), fibroblasts colony culture (CEU-F), hematopoiesis progenitor cell colony culture (CFU-GM, CFU-E, CFU-MK) and exogenous spleen colony-forming units count (CFU-S) were compared between two groups; on the basis of above indexes, the bone marrow histology and the capability of hematopoietic reconstruction of bone marrow cells were added on the sixth month after treatment.  RESULTS AND CONCLUSION: The general status of mice in the experimental group was better than that in the control group; in the third month, BMNC, CFU-GM and CFU-MK of the experimental group were higher than that of the control group, but no significant difference on the peripheral blood, CFU-F and CFU-E; on the sixth month, all the indexes mentioned above were higher than control group with exception of peripheral blood; during observation period all the indexes had a similar descending-tendency , but the tendency of experimental group was much slower than that of control group with significant difference (P < 0.05). Furthermore, the experimental group had more hematopoietic tissue but less adipose tissue than that of control group; The capacity of hematopoietic reconstruction of bone marrow cells from experimental group was better than that of control group. The results indicated that mPMSCs could delay the natural recession of mouse hematopoietic system. Related Articles | Metrics

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Clinical effects of haploidentical hematopoietic stem cell transplantation on high-risk leukemia Wan Ding-ming, Zhang Cheng, Xie Xin-sheng, Sun Hui, Sun Ling 2011, 15 (19):  3485-3488.  doi: 10.3969/j.issn.1673-8225.2011.19.015 Abstract ( 107 )   PDF (610KB) ( 478 )   Save BACKGROUND: Allogeneic hematopoietic stem cell transplantation is the main therapy of leukemia. The haploidentical hematopoietic stem cell transplantation can expand the application of the transplantation. OBJECTIVE: Observe the effect of “modi?ed Bu/Cy+ATG” conditioning regimen of haploidentical hematopoietic stem cell transplantation for patients with high-risk leukemia. METHODS: Nineteen patients with high-risk leukemia received a “modi?ed Bu/Cy+ATG” conditioning regimen: peripheral blood stem cells transplantation in 5 cases, and peripheral blood and bone marrow stem cells transplantation in 14 cases. Methotrexate, cyclosporine A, mycophenolate mofetil were used to prevent graft-versus-host disease (GVHD). Prostaglandin E1 was applied to prevent liver veno-occlusive disease (VOD). RESULTS AND CONCLUSION: ①Short term effect: the median neutrophil recovery time was 12 (8-20) d; the median platelet recovery time was 13 (10-31) d, and all the patients were successfully implanted. Transplant-related mortality rate was (15.8±8.4)% within 100 days after transplantation. ②GVHD incidence: the overall incidence of Ⅰ- Ⅳdegree of acute GVHD was (63.1±11.1)%, the incidence of chronic GVHD was (54.54±15.0)%. ③Long-term effect: 2-year disease-free survival rate was (28.2±15.5)%. 2-year overall survival rate was (46.9±16.5)%. For the patient with high-risk leukemia, who has not a HLA-matched related or unrelated donor but needs a salvage transplant, the haploidentical hematopoietic stem cell transplantation with “modi?ed Bu/Cy+ATG” conditioning regimen is a feasible selection. Related Articles | Metrics

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Umbilical cord mesenehymal stem cells transplantation for the immune system following systemic lupus erythematosus Xiao Yu-cui, Wang Ji-bo, Dong Jing, Gao Hong, Liang Hong-da, Wang Li, Xin Miao-miao 2011, 15 (19):  3489-3493.  doi: 10.3969/j.issn.1673-8225.2011.19.016 Abstract ( 120 )   PDF (1656KB) ( 578 )   Save BACKGROUND: Recently, many studies indicate that umbilical cord mesenehymal stem cells (UC-MSCs) have therapeutic effects on systemic lupus erythematosus (SLE). There are few reports, however, regarding the immunosuppressive effect and the organs of UC-MSCs on SLE .  OBJECTIVE: To investigate the efficacy and influence of UC-MSCs transplantation on immune systems and different organs in the treatment of SLE. METHODS: MRL/lpr female mice were divided into 4 groups: control group, cyclophosphamide (CTX) group, CTX+UC-MSCs group, and UC-MSCs group. 24-hour proteinuria was assessed using Bradford method．Enzyme-linked immunosorbent assay was used to measure the levels of serum anti-dsDNA antibodies and interleukin-10 (IL-10), IL-17, interferon gamma (IFN-γ), PDGF. The histopathology changes of the kidney were observed using hematoxylin-eosin staining, and the UC-MSCs surface marks CD105 and CD44 in the kidney, lung and spleen were observed. RESULTS AND CONCLUSION: ①The levels of IL-10, IL-17 and IFN-γ in the CTX+UC-MSCs group was lower than those in the control group (P < 0.05). ②At 2 weeks, the 24-hour proteinuria in the CTX+UC-MSCs and UC-MSCs groups was lower than that in the CTX and control groups (P < 0.01). ③The serum level of anti ds-DNA antibodies in the CTX+UC-MSCs group was significantly decreased as compared with that in the control group. ④After UN-MSCs transplantation, glomerulosclerosis and inflammatory cells infiltrating degree relieved. ⑤At 32 weeks, the expression of CD105 and CD44 in the kidney, lung and spleen was negative. The results indicated that MSCs transplantation can reduce lupus activity of MRL/lpr mice significantly and have marked therapeutic effect for lupus nephritis. It can also inhibit immune system by reducing the levels of inflammatory factors. Related Articles | Metrics

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Bone marrow mesenchymal stem cells prevent myocardial fibrosis via nuclear factor kappa B signaling pathway Deng Wei, Chen Qing-wei, Li Xing-sheng, Li Gui-qiong, Ke Da-zhi, Mo Xian-gang, Zan Li-ping 2011, 15 (19):  3494-3498.  doi: 10.3969/j.issn.1673-8225.2011.19.017 Abstract ( 147 )   PDF (1397KB) ( 550 )   Save BACKGROUND: Bone marrow mesenchymal stem cells (BMSCs) would show much help in treating myocardial fibrosis, and the mechanism involved remains unclear. OBJECTIVE: To investigate the efficacy and mechanism of myocardial fibrosis intervention using BMSCs. METHODS: BMSCs isolated from male mice were transfused into female mice with isoproterenol-induced myocardial fibrosis via tail vein (treated group). This study included three groups: treated, untreated, and control groups. Five weeks after transplantation, expression levels of sex-determining region of Y-chromosome (SRY), matrix metalloproteinases-9 (MMP-9) and tissue inhibitor of metalloproteinase-1 (TIMP-1) in myocardium were detected by fluorescent quantitative RT-PCR. Collagen distribution was observed using sirius red staining. The expression of nuclear factor-κB (NF-κB) was detected by immunohistochemistry. RESULTS AND CONCLUSION: BMSCs can home to fibroid heart. Compared to the untreated group, treated group had decreased levels of MMP-9, TIMP-1, NF-κB and collagen deposition (P < 0.05). The results show that BMSCs transplantation can improve MMP-TIMP expression and collagen deposition in injured myocardium after homing, a mechanism that may be related to the NF-κB mediated signaling pathway. Related Articles | Metrics

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Intravenous and local injection of umbilical cord blood progenitor cells for treatment of diabetic lower limb ischemia Peng Yan, Cheng Pei, Xu Yong 2011, 15 (19):  3499-3502.  doi: 10.3969/j.issn.1673-8225.2011.19.018 Abstract ( 119 )   PDF (1244KB) ( 396 )   Save BACKGROUND: Local intramuscular injection of endothelial progenitor cells (EPCs) is often used in the treatment of diabetic lower limb ischemia as well as animal experiments. OBJECTIVE: To compare the intravenous and local injection of umbilical cord blood progenitor cells effects on diabetic lower limb ischemia. METHODS: Wistar male rats were divided into five groups: ①Diabetic rats with tail intravenous EPC transplantation following ligation of bilateral hind femoral artery (diabetes+ligation+intravenous injection, DLV); ②Diabetic rats with local EPC transplantation into the right hind limb following ligation of bilateral hind femoral artery (diabetes+ligation+intramuscular injection, DLM); ③Normal rats with tail intravenous EPC transplantation following ligation of bilateral hind femoral artery (normal+ligation+intravenous injection, NLV); ④Diabetic rats with ligation of bilateral hind femoral artery (diabetes+ligation+control, DLC); ⑤Diabetic rats without transplantation and ligation (diabetes+control, DC). EPCs were traced with green fluorescence, the capillary number among muscle fibers was measured using hematoxylin-eosin staining, and RT-PCR detection was performed for vascular endothelial growth factor (VEGF) mRNA level. RESULTS AND CONCLUSION: Gastrocnemius ulcers and ischemia in the Right hind limb was obvious improved in the DLV group as compared with DLM group, but there was no significant difference. The expression of fluorescence in the DLV and DLM groups was similar. The capillary number was higher in the DLV group than the DLM group, but there was no significant difference. VEGF mRNA levels had no difference between the DLV and DLM groups (P > 0.05). The findings indicate that the therapeutic effects of umbilical cord blood progenitor cells via tail intravenous and local injection are similar. Related Articles | Metrics

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Olfactory ensheathing cell transplantation for treatment of sciatic nerve injury Yang Shuo, Hou Zhan-jiang, Xia Hao-chen, Fan Chang-dong 2011, 15 (19):  3503-3506.  doi: 10.3969/j.issn.1673-8225.2011.19.019 Abstract ( 69 )   PDF (1243KB) ( 385 )   Save BACKGROUND: Studies have demonstrated that olfactory ensheathing cells (OECs) are conducive to the survival of neurons and promote axonal regeneration. OBJECTIVE: To study the feasibility and effect of local injection of rat OECs to treat peripheral nerve injury. METHODS: Rat OECs were isolated and cultured in vitro. 1.0 cm sciatic nerve segment was cut from 40 adult female SD rats undergoing allograft nerve transplantation, and then the rats were randomly divided into two groups. OECs group: local injection of OECs; saline group: local injection of sterile saline. After 3 months, somatosensory evoked potentials (SEP) and motor evoked potentials (MEP) were detected and electrophysiological recovery of nerve was observed under light microscopy and electron microscopy. RESULTS AND CONCLUSION: The electron microscopy found that in the damage zone, OECs group had more nerve fibers significantly than saline group (P < 0.01); In OECs group, SEP and MEP latency and amplitude was significantly better than that in saline group (P < 0.01). it is shown that local injection of OECs can better restore function after peripheral nerve injury. Related Articles | Metrics

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Hyperbaric oxygen combined with neural stem cells transplantation for rat spinal cord injury Shi Rui-cheng, Feng Shi-jun, Wang Zhi-gang 2011, 15 (19):  3507-3512.  doi: 10.3969/j.issn.1673-8225.2011.19.020 Abstract ( 71 )   PDF (1421KB) ( 356 )   Save BACKGROUND: Neural stem cells (NSCs) transplantation is widely used in the repair of spinal cord injury (SCI). OBJECTIVE: To investigate the effect of NSCs transplantation plus hyperbaric oxygen (HBO) on repair of rat SCI.   METHODS: Sixty female rats were divided randomly into three groups: SCI group; NSCs transplantation group; NSCs transplantation+HBO group. Four weeks post-transplantation, histopathology examination and BrdU staining were performed. Eight weeks post-transplantation, horseradish peroxidase (HRP) nerve trace and transmission electron microscope testing and somatosensory evoked potential (SEP) testing were performed. At 1, 2, 4, 6, 8 weeks post-injury, all animals were evaluated for the hind limb behavior with Basso-Beattle-Bresnahan (BBB) score and inclined plane test.  RESULTS AND CONCLUSION: HRP-labeled neurofibra and BrdU positive cells were found in the spinal cord of the NSCs transplantation+HBO group, some in NSCs transplantation group but few in the SCI group spinal cord injury. SEP testing in the SCI group surpassed that in the NSCs transplantation group (P < 0.05). There was significant difference between spinal cord injury group and NSCs transplantation+ HBO group (P < 0.01). By transmission electron microscope, nerve fibers and myelinated nerve fibers were found in the central transverse plane in the NSCs transplantation group and NSCs transplantation+HBO group. The outcome of functional recovery in the NSCs transplantation+HBO group surpassed that in the NSCs transplantation group. There was significant difference between the NSCs transplantation and NSCs transplantation+ HBO groups (P < 0.05). The outcome of the SCI group was worse than that of the other groups. There was significant difference between the NSCs transplantation group, NSCs transplantation+ HBO group and SCI group (P < 0.05). NSCs transplantation can promote the functional recovery of rat hind limbs after SCI, and its combination with HBO has a synergistic effect. Related Articles | Metrics

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Transfusion support therapy for patients undergoing hematopoietic stem cell transplantation Song Kui, Qiu Da-fa, Guo Zi-wen, He Hui-qing, Jian Li, Huang Gui-nian, Huang Gui-ying, Liu An-li, Xu Xiao-jun 2011, 15 (19):  3513-3517.  doi: 10.3969/j.issn.1673-8225.2011.19.021 Abstract ( 65 )   PDF (1170KB) ( 274 )   Save BACKGROUND: Transfusion of blood products is often necessary for patients undergoing hematopoietic stem cell transplantation (HSCT). The need for red cell and platelet transfusion may vary significantly depending on the type of transplantation and underlying disease.OBJECTIVE: To evaluate the need and volume of transfusions in patients undergoing HSCT in Zhongshan Hospital of Sun Yat-sen University. METHODS: Retrospective data were collected on all patients under¬going autologous and allogeneic transplantation in our hospital during January 2004 to June 2010, regarding indications for transplantation, type of HSCT, CD34+ stem cell dose, total number of red cell and platelet units transfused, cost, and times to achieve freedom from transfusion (FFT) and engraftment for white blood cell and platelet. The trigger for red cell transfusion was hemoglobin lower than 70 g/L. Platelet count less than 20×109/L was the trigger for platelet transfusion. We evaluated the needs, doses as well as cost for red cell and platelet transfusion for patients undergoing HSCT. Also, overall and disease free survival were analyzed as clinical outcomes. RESULTS AND CONCLUSION: A total of 14 (93%) out of 15 patients undergoing autologous HSCT and 35 (90%) out of 39 patients with allogeneic HSCT exhibited total hematopoietic engraftment and FFT. Time to achieve FFT (median; range) for red blood cell units for autologous HSCT (14.6, 0-62) was significantly shorter compared with allogeneic HSCT (11.8, 0-108.5). Number of red blood cell units (median; range) transfused were significantly less in patients undergoing autologous HSCT (5.8, 0-35.5) compared to patients undergoing allogeneic HSCT (11.8, 0-108.5). The median cost of RBC transfusion was significantly higher in patients undergoing allogeneic HSCT compared to patients undergoing autologous HSCT．Transfusion of blood products is an expensive but integral part of HSCT, the more transfusion requirements for allogeneic HSCT than for patients undergoing autologous HSCT. FFT is a desirable long-term goal of successful HSCT. Related Articles | Metrics

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Effect of arsenic trioxide on multi-drug resistance of HL60/ADR cells Huang Ming, Meng Fan-yi, Cai Yan-xia, Wang Zhi-xiang, Xia Li-hua, Chen Jia-bin 2011, 15 (19):  3518-3521.  doi: 10.3969/j.issn.1673-8225.2011.19.022 Abstract ( 112 )   PDF (1173KB) ( 344 )   Save BACKGROUND: The treatment of arsenic trioxide (As2O3) on acute promyelocytic leukemia (APL) has acquired good effect. OBJECTIVE: To investigate the effect and mechanism of As2O3 on reversing multidrug resistance of HL60/ADR cell line. METHODS: HL60/ADR cells were divided into control group and As2O3 group. Cells of control group were incubated with nun-drug, those of As2O3 group incubated with 48-hour IC50 As2O3. RESULTS AND CONCLUSION: As2O3 could enhance the intracellular accumulation of DNR and decrease the expression of MRP1 in HL60/ADR cells (P < 0.05). At the same time, the early and late apoptosis rate of HL60/ADR cells both increased significant with time-dependently, which was higher than that in the control group (P < 0.01). As2O3 could also elevate the expression of bax, caspase-3, caspase-9, PARP, and decelerate the expression of Bcl-2, NF-κB p65, p-p65. Related Articles | Metrics

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Effect of pH levels in the survival and proliferation of adipose derived mesenchymal stem cells Wu Kun, Li Fang, Ma Jian, Li Jin-zhen 2011, 15 (19):  3522-3524.  doi: 10.3969/j.issn.1673-8225.2011.19.023 Abstract ( 95 )   PDF (775KB) ( 363 )   Save BACKGROUND: Decreased pH value has a great effect on adipose derived mesenchymal stem cells (ADSCs) in the degenerated intervertebral disc. OBJECTIVE: To observe the survival and proliferation of cultured ADSCs under different pH conditions.   METHODS: ADSCs were isolated from fat tissue of rabbits and cultured under four different pH conditions (pH 7.4, pH 7.1, pH 6.8, pH 6.5). Five days later, typan blue staining was used to detect cell viability and MTT method was applied to draw cell growth curve. RESULTS: The cell viability in the pH 6.5 group was the lowest compared with that in the other three groups (P < 0.05). It is indicated that pH 6.5 can inhibit the viability and proliferation of ADSCs. Related Articles | Metrics

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Cryopreservation of hematopoietic stem cells with ladder-style freezing from low temperature refrigerator to liquid nitrogen method versus programmed cooling method Zhao Ren-bin, Zhang Ai-ling, Lu Jie, Yang Ze-feng, Yang Tong-hua 2011, 15 (19):  3525-3527.  doi: 10.3969/j.issn.1673-8225.2011.19.024 Abstract ( 132 )   PDF (497KB) ( 618 )   Save BACKGROUND: During the cryopreservation of hematopoietic stem cells, there are many influential factors, such as cooling rate, storage temperature, and cryoprotectant combination. There is a controversy in the cryopreservation methods. OBJECTIVE: To investigate the differences between -80 ℃ ladder-style freezing from low temperature refrigerator to liquid nitrogen method and programmed cooling method in preserving peripheral blood stem cells (PBSCs). METHODS: Collected PBSCs were divided into two groups, and respectively cryopreserved by using -80 ℃ ladder-style freezing from low temperature refrigerator to liquid nitrogen method and programmed cooling method. RESULTS AND CONCLUSION: There was no significant difference between these two frozen samples of PBSCs after recovery in the aspects of trypan blue exclusion rate, recovery, apoptosis rate and mortality (P > 0.05). These findings indicate that the -80 ℃ ladder-style freezing from low temperature refrigerator to liquid nitrogen method using the cryoprotectant containing 5% dimethyl sulfoxide, 4% albumin and 3% hydroxyethyl starch can yield the same effect as same as the programmed cooling method using 10% DMSO as cryoprotectant in the aspect of PBSCs cryopreservation, and the former one is more convenient and more suitable for clinical application. Related Articles | Metrics

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Application of cold enzyme digestion in isolating and culturing Schwann cells from newborn rats Mei Yu-feng, Huang Min, Yan Yu-hua 2011, 15 (19):  3528-3530.  doi: 10.3969/j.issn.1673-8225.2011.19.025 Abstract ( 80 )   PDF (503KB) ( 452 )   Save BACKGROUND: Currently, conventional enzymatic digestion methods are commonly used to isolate and culture Schwann cells. Because trypsinase has toxic effects on Schwann cells in 30 minutes at 37 ℃, the amount and purity of Schwann cells can have a certain impact. OBJECTIVE: To improve a novel method to rapidly isolate and culture a great amount of Schwann cells. METHODS: Schwann cells were isolated from newborn rat sciatic nerve digested with trypsinase at 4 ℃, 6 to 18 hours, and then cultured for some time. MTT was used to draw cell growth curve. Hematoxylin-eosin staining and immunohistochemistry were used to observe cell morphology, and positive cells were counted. RESULTS AND CONCLUSION: The quantity of Schwann cells was over 1×106 and the purity of Schwann cells was over 90% assessed by anti-s100 protein immunohistochemistry stainer. From the morphology of cultured cells and cells growth curve, the cold enzyme digestion method was simply and effective, the purified cells had good biologic characteristics. In conclusion, the novel method can satisfy the clinical need such as isolating and culturing time, the quantity and purity of cells and the bioactivity of Schwann cells. Related Articles | Metrics

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The development of device for rotary three-dimensional cells culture  Zhao Ling-xi, Ding Hao, Xu Meng, Shang Kun, Shen Li-xing 2011, 15 (19):  3531-3533.  doi: 10.3969/j.issn.1673-8225.2011.19.026 Abstract ( 117 )   PDF (531KB) ( 310 )   Save BACKGROUND: It is widely used that vitro culturing endothelial cells to study atherosclerosis mechanism, and the key issues are if the monolayer endothelial cells could resist blood attack and how to achieve cells growth well-distributed. The development of this device is aim to solve these problems. OBJECTIVE: To develop a device that could supply a three-dimensional condition for cells to realize growth stick to elastic silicon chamber in order to improve growth uneven. METHODS: It Weintroduced structure and functions of the device from controlled unit, mechanical unit and cells culture unit, and then used it to carry out an umbilical vein endothelial cells culture experiment. RESULTS AND CONCLUSION: Rotary three-dimensional cells culture device could provide condition for cells metabolism and growth sticking to elastic silicon chamber well-distributed. Related Articles | Metrics

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Effect of olfactory ensheathing cells transplantation on the expression of transforming growth factor beta in the motor cortex of rats subjected to spinal cord transection Li Yun, Peng Chun, Wang Ting-hua 2011, 15 (19):  3534-3537.  doi: 10.3969/j.issn.1673-8225.2011.19.027 Abstract ( 92 )   PDF (574KB) ( 381 )   Save BACKGROUND: Many studies have showed that olfactory ensheathing cells (OECs) transplantation can result in significant functional improvements in rats subjected to spinal cord injury, but the underlying molecular mechanisms is not unknown. OBJECTIVE: To observe the effect of OECs transplantation on the expression of transforming growth factor beta (TGF-β) mRNA in the motor cortex of the rats subjected to spinal cord transection. METHODS: OECs isolated from transgenic mice expressing GFP and cultured by enzymatic digestion were made into cell suspension. Model of spinal cord transection (SCT) was established by cut transversely Sprague-Dawley (SD) rats T9 segment. After modeling, the rats were divided into sham-operation group, operation group and OECs transplantation group. RT-PCR was employed to detect gene expression of TGF-β. β-actin was used as control. RESULTS AND CONCLUSION: RT-PCR results showed that compared with sham-operation group, the gene expression of TGF-β in the motor cortex increased significantly at 3 days postoperatively in the operation group (P < 0.05), and it returned to the level of sham-operation group at 7, 14, 21, and 28 days after modeling. Following OECs transplanatation, the expression of TGF-β in motor cortex at 21 days postoperatively showed a significant decrease compared with the operation group (P < 0.05). The findings indicate that spinal cord transection results in the up-regulation of the expression of TGF-β mRNA in the motor cortex in early period. OECs engrafted can decrease the expression of TGF-β mRNA in the later time so as to promote the recovery of spinal cord injury. Related Articles | Metrics

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Culture and purification of olfactory ensheathing cells from rats using cytarabine combined with nerve growth factor Liu Xiao-yang, Sun Jian-min, Cui Xin-gang, Jiang Zhen-song 2011, 15 (19):  3538-3541.  doi: 10.3969/j.issn.1673-8225.2011.19.028 Abstract ( 124 )   PDF (1548KB) ( 274 )   Save BACKGROUND: Because of the continual capcity of myelination, olfactory ensheathing cells (OECs) are preferred by numerous researchers. The first step is to gain a great quantity of purified OECs simply and economically. The following provides a feasible method to culture OECs. OBJECTIVE: To explore a simple and pragmatic method to obtain sufficient OECs from neonatal rats by using cytarabine combined with nerve growth factor (NGF). METHODS: After different attachment, DMEM/F12 culture solution including 10 mg/L cytarabine was used to culture OECs for 48 hours. Then, OECs were cultured in DMEM/F12 solution with 25 μg/L NGF and 1% fetal bovine serum. The conditions and growth degree of OECs were observed and P75 immunocytochemistry was used to estimate the cell purity. RESULTS AND CONCLUSION: OECs were positive after P75 immunocytochemistry. They appeared to be bipolar or tripolar cells and their processes formed a network in vitro. The purity of OECs in good conditions reached about 95% on 7 days and 90% on 9 days. The method of using cytarabine combined with NGF can culture and purify OECs simply and effectively. Related Articles | Metrics

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Three-dimensional reconstruction and display of embryoid body-like structure Shen Xiao-tao, Zheng Xin, Zhong Jia-yong, Zhang Zhi-peng, Li Zhen, Cai Dong-qing 2011, 15 (19):  3542-3545.  doi: 10.3969/j.issn.1673-8225.2011.19.029 Abstract ( 67 )   PDF (1093KB) ( 285 )   Save BACKGROUND: Stem cells have been widely studied and are considered as important cells for cell therapy since stem cells are capable of proliferation and differentiation. It is believed that three-dimensional (3D) reconstruction and display of stem cells and tissues is important to better understand the mechanisms of proliferation and differentiation of stem cells. OBJECTIVE: To reveal and analyze embryoid body-like tissue structure using 3D-reconstruction technique. METHODS: Differentiated embryoid body-like tissue structure was used as a model. Immunofluorescent staining combined with Z-axis consecutive image capture and 3D-reconstruction were used to reveal and analyze differentiated embryoid body-like cardiac myocyte tissue. RESULTS AND CONCLUSION: Methods used in this study were able to reveal 3D-reconstruction of embryoid body-like tissue structure. Reconstructed 3D model could be rotated at any angle. After iso-surface processing under certain color channel, spatial relationship of cardiac troponin I positive tissue structure and nucleus could be displayed on related color channel or at any angle dimension. It was found that the nuclei were included in the differentiated embryoid body-like cardiac myocyte tissue. The results of present study documented that differentiated embryoid body-like cardiac myocyte tissue was able to reveal and analyze using the 3D-reconstruction technique. Related Articles | Metrics

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Distribution of Telocytes in the rat heart Liu Juan-juan, Shen Xiao-tao, Zheng Xin, Li Zhen, Wang Jian, Qi Xu-feng, Cai Dong-qing 2011, 15 (19):  3546-3548.  doi: 10.3969/j.issn.1673-8225.2011.19.030 Abstract ( 229 )   PDF (712KB) ( 545 )   Save BACKGROUND: Telocytes in the heart are possible to act as linker and supporting cells between cardiac myocytes and endogenous cardiac stem cells, and play an important role in myocardial regeneration and ventricular remodeling after myocardial infarction (MI). However, the distribution of Telocytes in the heart and their functional role are still unclear. OBJECTIVE: To investigate the distribution of Telocytes in the rat heart. METHODS: The continuous frozen sections were conducted using female SD rat heart. The representation sections from the base, the medium, and the atrium-atria of the heart were selected for immunofluorescence staining with c-kit antibody. The c-kit positive and unique morphology of Telocytes were applied as standard to identify the distribution of Telocytes in the rat heart. RESULTS AND CONCLUSION: The cell density of Telocytes in the base part (26 cells/mm2) and the atrium-atria part (20 cells/mm2) was significantly higher than that in the medium part (9 cells/mm2). In addition, the cell density of Telocytes in the subepicardium (the base: 18 cells/mm2; the medium: 5 cells/mm2; the atrium-atria: 13 cells/mm2) was significantly higher than that in the endocardium (the base: 7 cells/mm2; the medium: 4 cells/mm2; the atrium-atria part: 7 cells/mm2). The above-mentioned results suggested that the distribution of Telocytes in the heart is different. The pattern of Telocytes distribution in the heart may be related to the different distribution of cardiac myocytes and endogenous cardiac stem cells. Related Articles | Metrics

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Biological characteristics and multilineage differentiation of superparamagnetic iron oxide nanoparticles labeled adipose tissue-derived stem cells from pigs and in vitro 3.0T MR imaging Wang Ji-fei, Jiang Li, Yang You-you, Wang Li-qin, Cai Hua-song, Zhao Jing, Li Zi-ping, Yang Jian-yong 2011, 15 (19):  3549-3554.  doi: 10.3969/j.issn.1673-8225.2011.19.031 Abstract ( 115 )   PDF (1808KB) ( 504 )   Save BACKGROUND: Adipose tissue-derived stem cells (ADSCs) are promising for clinical application in cardiovascular cellular therapy. In vivo MR imaging of magnetically labeled ADSCs to track the survival and migration of implanted cells is important for clinical application and scientific research. However, to carefully evaluate the safety of any labeling method and investigate the characteristics of in vitro MR imaging is required before any clinical application. OBJECTIVE: To label porcine ADSCs with superparamagnetic iron oxide nanoparticles (SPIO) combined with poly-l-lysine (PLL), and to explore the effects of magnetic cell labeling on cellular biological characteristics, multilineage differentiation potential, and the characteristics of in vitro 3.0T MR imaging. METHODS: ADSCs were isolated from the subcutaneous adipose tissue of pigs and expanded in culture medium. ADSCs were incubated with the SPIO-PLL complex for 48 hours. Cell viability, proliferation and differentiation capacity to cardiomyocytes, osteogenic and adipogenic lineage between labeled and unlabeled cells were compared. Different concentrations of labeled ADSCs and the control were scanned at a clinical 3.0T MR system by using T1WI, T2WI and T2*WI sequences. RESULTS AND CONCLUSION: Numerous intracytoplasmic iron particles stained with Prussian blue were detected in labeled cells with labeling efficiency of nearly 100%. Both labeled and unlabeled cells were able to differentiate into cardiomyocytes, adipocytes and osteocytes. The change in MR signal intensity increased at T1WI, T2WI and T2*WI along with elevation of the labeled cells concentration. The biggest and smallest change in signal intensity were observed at T2*WI and T1WI respectively. There were significant differences among the change in signal intensity at T1WI, T2WI and T2*WI for each cell concentration (P < 0.01). At least 1×106/L SPIO labeled ADSCs could be detected by 3.0T MR. Our study indicated that ADSCs are more promising cells for clinical application in repairing damaged myocardium. ADSCs can be effectively labeled by SPIO-PLL complex without affecting the cell viability, proliferation and multilineage differentiation ability. Compared with T1WI and T2WI, T2*WI is the most sensitive sequence for MR tracking of SPIO labeled ADSCs. 3.0T MR is more sensitive for cellular tracking based on the advantage of higher magnetic field strength. Related Articles | Metrics

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Stem cell characteristics of human amniotic epithelial cells Jin Ling, Chen Jian, Wu Jing, Xu Jin-tang, Zhou Qing, Liu Xiao-yong 2011, 15 (19):  3555-3558.  doi: 10.3969/j.issn.1673-8225.2011.19.032 Abstract ( 160 )   PDF (847KB) ( 409 )   Save BACKGROUND: Studies have found that human amniotic epithelial cells (hAECs) have a multiple differentiation capacity similar as pluripotent stem cells or embryonic stem cells. It is indicated that hAECs may be a new kinds of seed cells in tissue engineering reconstruction in the further. OBJECTIVE: To culture the hAECs in vitro and to investigate stem cell characteristics of cultured hAECs. METHODS: The human amniotic tissue was delivered from full term caesarean, and the highly purified hAECs were obtained from enzyme digestion method and differently sticky method. The hAECs were seeded in DMEM/F12 medium obtaining 10% fetal bovine serum for the first time and serial subcultivation. The immunofluorescence and flow cytometry were used to detect the expression of OCT-4 of stem cell factor and Stem cell surface molecule marker CD29, CD34, CD44, CD45, CD105 in passaged hAECs. RESULTS AND CONCLUSION: The cells in vitro looked like slabstone that epithelial cells specified. The results of immunofluorescence showed the expression of OCT-4 of stem cell factor in hAECs was positive. Detection of stem cell surface molecule marker by the flow cytometry showed the expression of CD29, CD34 on hAECs were positive, and the expression of CD44, CD45, CD105 on hAECs were negative. The findings show that the hAECs have some characteristics of embryonic stem cells, and may be a new kind of seed cells for the future tissue engineering reconstitution. Related Articles | Metrics

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Directed differentiation of ectomesenchymal cells isolated from the first branchial arch into adipocytes Hou Jing-qiu, Yan Zheng-bin, Yang Xu 2011, 15 (19):  3559-3562.  doi: 10.3969/j.issn.1673-8225.2011.19.033 Abstract ( 96 )   PDF (1210KB) ( 319 )   Save BACKGROUND: During the dentofacial development, the first branchial arch, as a transitional structure, plays an important role. OBJECTIVE: To investigate the self-renewal of ectomesenchymal cells (EMCs) isolated from the first branchial arch as well as the feasibility of inducing EMCs into adipocytes. METHODS: The activity of proliferation and self-renewal of EMCs were detected by nuclei labeled with BrdU and fluorescence in situ hybridization for telomerase. Monolayer cultures of EMCs were passaged 3 times and then transferred to adipogenic media. The level of differentiation was evaluated by histological examination and the expression of tissue-specific genes was analyzed by reverse transcription-polymerase chain reaction technique. RESULTS AND CONCLUSION: Nuclei labeled with BrdU and fluorescence in situ hybridization for telomerase were positive. After being cultured in adipogenesis-inducing medium, EMCs responded by the accumulation of lipid vacuoles and the gene expression of LPL. The findings indicate that EMCs have a high activity of proliferation and self-renewal, and they are capable of differentiation into adipocytes in vitro. Related Articles | Metrics

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Effect of pilose antler polypeptides on chondrogenic phenotype differentiation of bone marrow-derived mesenchymal stem cells in vitro Xiu Zhong-biao, Lin Jian-hua, Wu Zhao-yang, Wang Ri-xiong 2011, 15 (19):  3563-3566.  doi: 10.3969/j.issn.1673-8225.2011.19.034 Abstract ( 83 )   PDF (411KB) ( 327 )   Save BACKGROUND: Pilose antler polypeptides (PAP) have been proved to promote the proliferation of condrocytes cultured in vitro and expressions of glycosaminoglycan (GAG), type Ⅱ collagen and Aggrecan protein in the extracellular matrix. OBJECTIVE: To investigate the feasibility of chondrogenic phenotype differentiation of rabbit bone marrow-derived mensenchymal stem cells (BMSCs) in a defined medium and then to study the effect of PAP on chondrogenic differentiation of BMSCs in vitro. METHODS: The third passage BMSCs from rabbits were randomly divided into control group cultured in ordinary medium, induced group cultured in defined medium, and PAP group cultured in defined medium containing 10 mg/L PAP. An equal volume of articular chondrocytes were selected from rabbits as articular cartilage group. The cellular morphological and functional characteristics were observed after 1, 2, 3 weeks in centrifuge tubes by histological, biochemical and reverse transcription-polymerase chain reaction (RT-PCR) technique. RESULTS AND CONCLUSION: Cell masses in the control group gradually crumbled after 2 weeks, and hematoxylin-eosin staining could not be done. Cell masses in the induced and PAP groups were semitransparent, but slightly contracted. A part of these cells were round or oval with a high density distribution at the surface. The content of GAG and mRNA expression of type Ⅱ collagen in the induced and PAP groups were increased with culture time, and higher than those in the control group at different time points (P < 0.05). The content of GAG and mRNA expression of type Ⅱ collagen in the PAP group were higher than those in the induced group, but lower than those in the articular cartilage group (P < 0.05). The results indicated that BMSCs can differentiate into chondrogenic phenotype in the defined medium, and PAP can significantly enhance chondrogenic phenotype differentiation of BMSCs. But the quality of cultured cartilage tissue is poorer than that of the articular cartilage. Related Articles | Metrics

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Effects of glucose concentration on differentiation of rat pancreatic duct stem cells Chen Hao-qiang, Shi Guang-jun, Xu Ping, Wu Xiao-ping 2011, 15 (19):  3567-3570.  doi: 10.3969/j.issn.1673-8225.2011.19.035 Abstract ( 99 )   PDF (339KB) ( 375 )   Save BACKGROUND: Glucose is an important factor on differentiation of pancreatic duct stem cells, it relates to the quantity and secretion function of insulin-producing cells after differentiation. OBJECTIVE: To compare the insulin secretion capacity of the differentiated rat pancreatic stem cells induced by various glucose concentrations. METHODS: Rat stem cells were isolated and purified from pancreatic duct cells using collagenase V and Ficoll-400. These stem cells were randomly divided into 10 groups. Every group was induced to culture, proliferate, differentiate and form insulin- producing cells in vitro. The differentiation of all groups was performed in medium with different concentrations of glucose. The immunofluorescence staining was used to identify the pancreatic duct stem cells. The electrochemical luminescence method was used to detect the insulin release from stem cell differentiated islets. RESULTS AND CONCLUSION: The stimulation index of glucose 20.6, 25.6, 30.6 mmol/L groups was higher than that in other groups (P < 0.05), but there was no difference between each two groups among these three groups (P > 0.05). The insulin releasing of glucose 15.6, 20.6, 25.6 groups was higher than that in other groups (P < 0.05), but there was no difference between each two groups among these three groups (P > 0.05). The best insulin secretion capacity of insulin-producing cells can be gained by controlling concentration of glucose as 20.6-25.6 mmol/L when pancreatic duct stem cells differentiated into insulin-producing cells. Related Articles | Metrics

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Migration mechanism of bone marrow mesenchymal stem cells Xu Shi-hong, Xie Xing-wen 2011, 15 (19):  3573-3576.  doi: 10.3969/j.issn.1673-8225.2011.19.037 Abstract ( 77 )   PDF (625KB) ( 391 )   Save BACKGROUND: Based on the multiple differentiation potential, bone marrow mesenchymal stem cells (BMSCs) migrating to the injury site greatly promote patient’s rehabilitation. OBJECTIVE: To investigate the mechanism of cytokines and inflammatory factors to promote the migration of BMSCs based on analysis of related literature. METHODS: The first author retrieved China National knowledge Infrastructure and PubMed datebases for concering articles .The key words were “BMSCs, chemotactic factor, migration” in Chinese and English. A total of 68 articles were retrieved primarily by computer, and 23 articles were finally included for review according to inclusion criteria.  RESULTS AND CONCLUSION: BMSCs have a strong differentiation potential and play a huge assistance role in the clinical treatment. At the same time, because BMSCs are easy and simple to harvest and culture in vitro, studies concerning the proliferation, differentiation and migration of BMSCs are hot points. The intervention function of related cell factors urges BMSCs to strength migration. Related Articles | Metrics

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Current understanding into the biological characteristics of mesenchymal stem cells Lin Zhi-xiang, Xu Hao 2011, 15 (19):  3577-3580.  doi: 10.3969/j.issn.1673-8225.2011.19.038 Abstract ( 104 )   PDF (569KB) ( 445 )   Save BACKGROUND: Mesenchymal stem cells (MSCs) have been widely used in clinical studies and treatment. Numerous studies have reported beneficial effects of MSCs in tissue repairing as well as immunomodulatory function reported recently. But it is unclear that how MSCs exert the effects. OBJECTIVE: To discuss the immunomodulatory mechanism of MSCs. METHODS: The PubMed database was researched using the computer for articles published from 2000 to 2010 using key words “mesenchymal stem cells, tissue repairing, homing, immunomodulatory” in English. Inclusion criteria: articles relevant to the machnism of MSCs to exert homing and immunomodulatory effects after tissue injury. Exclusion criteria: repetitive studies or irrelevant articles. RESULTS AND CONCLUSION: Totally 50 articles were included in the result analysis. MSCs effects on the repair of tissue injury have been confirmed, and the immune function of MSCs has been reported recently. The mechanism is pleiotropic and redundant, mediated by multiple factors glands - receptor reaction and related immune cells which are involved in this process. The mechanisms of MSCs are still unclear, further researches are needed to confirm and improve, so as to further promote clinical research and treatment. Related Articles | Metrics

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Effect of stem cells and strategy of traditional Chinese medicine in cardiac regenerative medicine  Yao Kui-wu, Wang Jie, Zhang Liang-deng 2011, 15 (19):  3581-3585.  doi: 10.3969/j.issn.1673-8225.2011.19.039 Abstract ( 98 )   PDF (703KB) ( 444 )   Save BACKGROUND: Traditional Chinese medicine (TCM) has its unique understanding on cardiac regeneration, but there is preliminary exploration stage of stem cells and cardiac regenerative medicine intervention in TCM. OBJECTIVE: To analyze and summarize correlative research development of stem cells and cardiac regeneration, and preliminarily discuss how to intervene cardiac regeneration in TCM. METHODS: PubMed database was searched with the key words “stem cell, cardiac regenerative medicine” and China National Knowledge Infrastructure with the key words “stem cell, TCM” for literatures published between 2000 and 2010, no limitation in language. We analyzed and summarized the literatures evidence that met criterions. Finally, 43 articles were included. RESULTS AND CONCLUSION: The analysis demonstrated that the study of use of stem cells in cardiac regenerative medicine has been expanded widely, including embryonic stem cells, bone marrow stem cells, adult cardiac stem cells, et al, which indicate the ability to regenerate myocardium and application perspectives. The understanding on cardiac regeneration in TCM is contained in yin and yang concept, the law of mother and son in five elements and vital essence theory. TCM needs to collate the understanding of cardiac regenerative medicine in TCM theory systematically, TCM compounds, value active components of TCM targeting to stem cells and to enhance researches on mechanisms of effective TCM. Related Articles | Metrics

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Progress of adipose-derived stem cells in orthopedics research Chen Lei-jie, Wang Wei, Dang Xiao-qian, Wang Kun-zheng 2011, 15 (19):  3586-3589.  doi: 10.3969/j.issn.1673-8225.2011.19.040 Abstract ( 120 )   PDF (673KB) ( 359 )   Save BACKGROUND: Adipose-derived stem cells (ADSCs) have become increasingly popular since it was discovered, which support a new way to treat orthopedic diseases. OBJECTIVE: To review the biological characterization and progress of ADSCs in orthopedic basic research, and provide powerful theoretical basis for further basic researches. METHODS: The first author retrieved ISI Web of knowledge database (http://www.isiknowledge.com) and PubMed database (http://www.ncbi.nlm.nih.gov/PubMed) for articles concerning the biological characterization of ADSCs and restoration of orthopedic diseases extensively published from 2001 to 2010. The key words were “adipose-derived stem cell，orthopaedic”. RESULTS AND CONCLUSIONS: ADSCs have similar characterization and multipotential differentiation as bone marrow-derived stem cells. Meanwhile, ADSCs reveal excellent results in defect repair of bone, cartilage, intervertebral disk, tendon and ligament. Furthermore, because of the abundance and ease of harvest, ADSCs can support adequate seed cells for cell engineering, tissue engineering and medicine researching. Therefore, ADSCs have manifested broad prospects in therapy of orthopedic diseases, which deserve a further investigation. Related Articles | Metrics

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Research progress on induced efficiency and methods of induced pluripotent stem cells Zhang En-huan, Hu ZHi-xing, Li Ma-lin 2011, 15 (19):  3590-3593.  doi: 10.3969/j.issn.1673-8225.2011.19.041 Abstract ( 132 )   PDF (529KB) ( 589 )   Save BACKGROUND: Induced pluripotent stem cells (iPS) is a type of pluripotent stem cell artificially derived from human or animal adult somatic cells which were transfected with certain pluripotency genes. At present, the researches on induced pluripotent stem cells have made great progresses. OBJECTIVE: To review the research progress in the conversion efficiency and optimization of induction methods in iPS cells. METHODS: A computer-based online search of Medline database (http://www.ncbi.nlm.nih.gov/Pubmed) from January 2006 to July 2010 was performed with the key words of “induced pluripotent stem cells” in English. The literatures were limited to English language. Totally 1 831 literatures were retrieved, 57 documents of which were involved for summarization. RESULTS AND CONCLUSION: The researches on iPS cells have a great and far-reaching impact on life science, which makes it become a hot issue in stem cell field. And some rapid progresses have been made in iPS research such as cellular reprogramming, the improvement of efficiency of induction, optimization of the induction methods, the enlargement of cell origin and differentiation of functional cells. These researches would lay a foundation for clinical cell replacement therapy of iPS. Related Articles | Metrics

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Traditional Chinese medicine promotes proliferation and differentiation of mesenchymal stem cells Wang Yan-chun, Qu Jing, Han Ya-ling 2011, 15 (19):  3594-3597.  doi: 10.3969/j.issn.1673-8225.2011.19.042 Abstract ( 163 )   PDF (575KB) ( 719 )   Save BACKGROUND: Mesenchymal stem cells are the hotspot in regenerative medicine research, and traditional Chinese medicine (TCM) can improve the efficacy of mesenchymal stem cells therapy. OBJECTIVE: To review the correlation between TCM and stem cells, and to further explore their development trend. METHODS: A computer retrieve of CNKI and PubMed databases using the key words of which were “traditional Chinese medicine” or “herbal medicine” or “Botanical Drugs” and “mesenchymal stem cells” in Chinese and English was performed. RESULTS AND CONCLUSION: Totally 35 articles were included. The results showed that many Chinese herbs and compound play a role in improving the proliferation and differentiation of mesenchymal stem cells, which is safe and easy to clinical application. In addition, Chinese herbs have a great potential to survive in mesenchymal stem cells. Related Articles | Metrics

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Evidence-based treatment for incomplete spinal cord injury induced paraplegia in 1 case  Pan Ping, Yang Qi, Liang Qing-le 2011, 15 (19):  3598-3601.  doi: 10.3969/j.issn.1673-8225.2011.19.043 Abstract ( 142 )   PDF (506KB) ( 434 )   Save BACKGROUND: It is difficult to cure spinal cord injuries (SCI) in the world. The injured neuron can not renew in SCI. The treatment is different in every hospital, but there is not effective evidence based medicine (EBM) to support it. OBJECTIVE: To choice the best effective treatment target and method depending on the method of EBM for one SCI patient. METHODS: We searched the Cochran library, Pubmed, and Embase databases after fully acquiring the ordinary circumstances about the patient. The key words were “spinal cord injuries; pharmacotherapy; long-term; rehabilitation; prognosis-assessment; cell transplantation therapy; Prevention-of-complications; Meta-analysis; human; RCT; systematic review”. Twenty-seven articles were included in the result analysis. RESULTS AND CONCLUSION: The most important aim was to complications prevention and rehabilitation treatment, especially to promote the regeneration and function rehabilitation of the spinal cord as possible as we could. After half a year, the patient’s situation was stable and self-defecation, muscle strength of the upper limb was restored and lower limb spasticity relieved. The patient could self-stand. It is proved that EBM is proper to the situation of the patient. The method of EBM can make good results in clinic when it is selected as the treatment to SCI patients. Related Articles | Metrics

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Influences of different cryoprotectants by ladder-style freezing from low temperature refrigerator to liquid nitrogen on cryopreservation of hemopoietic stem cells Yang Tong-hua, Zhao Ren-bin, Jiang Ya-xian, Ouyang Hong-mei, Zhang Ai-ling, Hu Peng, Liu Jian-qiong, Wen Yan 2011, 15 (19):  3602-3606.  doi: 10.3969/j.issn.1673-8225.2011.19.044 Abstract ( 125 )   PDF (643KB) ( 295 )   Save BACKGROUND: Uncontrolled-rate freezing in -80 ℃ refrigerator is convenient, while controlled-rate freezing in -196 ℃ liquid nitrogen is reliable and long-term, the combination of the two can simplify the process and has been successfully used in clinics. OBJECTIVE: To explore the influences of different cryoprotectants by ladder-style freezing from -80 ℃ low temperature refrigerator to liquid nitrogen on the cryopreservation of hemopoietic stem cells. METHODS: The experiments were divided into four groups: 10% dimethyl sulfoxide (DMSO) group, 5% DMSO combined with 3% hydroxyethyl starch group, 5% DMSO combined with 0.25 mol/L trehalose group, 5% DMSO combined with 3% hydroxyethyl starch and 0.25 mol/L trehalose group. Peripheral hemopoietic stem cells were cryopreserved by ladder-style freezing from -80 ℃ low temperature refrigerator to liquid nitrogen. The ultrastructural changes were examined by transmission electron microscopy, the expressions of Annexin-V, PI and Caspase-3 were detected by flow cytometry. RESULTS AND CONCLUSION: There was no significant difference in survival rate, apoptotic rate and necrotic rates of the cryopreserved cells in the four groups (P > 0.05). The ultrastructural changes had no significant difference under the transmission electron microscopy. The viability was more than 90% in frozen-thawed mononuclear cell colonies, and the apoptosis was roughly 50% in the frozen-thawed CD45+ cell population, which contained many mature cells. Of hemopoietic stem cells, early stage cells have greater resistance to damage of cryopreservation than late stage cells. It is concluded that the addition of hydroxyethyl starch or trehalose into DMSO exhibits no synergistic protective effect on the cryopreservation of hemopoietic stem cells. Related Articles | Metrics

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Effects of electromagnetic field activated-ERK signaling pathway on proliferation and osteogenic differentiation of rat bone marrow mesenchymal stem cells Yang Guo-hua, Wu Hua, Zhao Dong-ming 2011, 15 (19):  3607-3610.  doi: 10.3969/j.issn.1673-8225.2011.19.045 Abstract ( 80 )   PDF (347KB) ( 433 )   Save BACKGROUND: It has been demonstrated that electromagnetic field (EMF) can adjust proliferation and differentiation of bone marrow mesenchymal stem cells, but the specific mechanism is not clear. OBJECTIVE: To investigate the effects of EMF-activated ERK1/2 pathway on proliferation and osteogenic differentiation of rat bone marrow mesenchymal stem cells. METHODS: The 3rd passage of rat bone marrow mesenchymal stem cells were received EMF treatment (15 Hz, 1 mT, sine wave), 20 μmol/L PD98059 + EMF treatment, or only PD98059 treatment. Simultaneously, a normal control group was established. Western blotting was applied to detect the activation of ERK signal pathway after EMF exposure. MTT assay was used to determine the activation of proliferation of cells. And alkaline phosphatase (ALP) activity in cells was detected by an ALP kit. RESULTS AND CONCLUSION: The ERK1/2 phosphorylation, proliferation and ALP activity of rat bone marrow mesenchymal stem cells were remarkably increased after exposure to EMF (P < 0.01). PD98059 could effectively block the increasing of ERK1/2 phosphorylation and cell proliferation (P < 0.01), but elevate ALP activity in a certain level (P < 0.01). EMF stimulation can fast activate ERK1/2 signal pathway and then promote the proliferation of rat bone marrow mesenchymal stem cells, however, ERK1/2 signal pathway activation has a less effect on osteogenic differentiation of bone marrow mesenchymal stem cells. Related Articles | Metrics