1.Isolation and purification of effective components in Plumula Nelumbinis by macroporous adsorption resins
Jixiang ZHANG ; Dongran FENG ; Lailiang OU
Chinese Traditional and Herbal Drugs 1994;0(12):-
Objective To investigate the adsorption function of macroporous adsorption resins for the separation and purification of the effective components in Plumula Nelumbinis. Methods AKS-W, AB-8, and H214 resins were systematically studied for their adsorption capability, and the contents of the liensinine, isoliensinine, and neferine were determined by HPLC. Results AKS-W Resin had a preferable adsorptive and separateive effect for the effective components in Plumula Nelumbinis. The products containing 10.2% liensinine, 6.7% isoliensinine, and 11.9% neferine could be obtained. Conclusion Macro-porous-type and non-polar adsorption resin AKS-W is an ideal adsorbent for the extraction of the effective components in Plumula Nelumbinis.
3.Genetic modification of bone marrow mesenchymal stem cells with human CXCR4 gene and migration in vitro.
Yue ZHANG ; Lailiang OU ; Zhaokang CHENG ; Xiaohua JIA ; Nianfa GAO ; Deling KONG
Journal of Biomedical Engineering 2009;26(3):595-600
This study was amied to construct CXCR4 gene modified bone marrow mesenchymal stem cells (MSCs), and investigate the effect of CXCR4 expression on MSCs migration. The retrovirus vector pMSCV-CXCR4-IRES-GFP that expresses human CXCR4 gene was cloned,the MSCs were transduced by the virus, and the expression of OXCR4 was analyzed by FACS, RT-PCR and immunofluorescence staining. The migration assay was performed using Transwell method in the presence of SDF-1. FACS results showed that 46% of the transduced MSCs were CXCR4 positive, and 57% were GFP positive. The expression of CXCR4 in MSCs was also confirmed by RT-PCR and immunostaining. The migration of MSCs was induced by SDF-1 and was strongly dependent on CXCR4 expression. The concentration of SDF-1 had effect on the migration and the transmigration rate of CXCR4 modified; the amount of MSCs was 5-fold higher than that of untransduced MSCs when the optimal concentration rose to 50 ng/ml. These data indicate that SDF-1/CXCR4 plays an important role in MSCs migration ,and the CXCR4 genetic modification approach could be applied to enhance cell homing, and engraftment in MSCs therapy.
Bone Marrow Cells
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cytology
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Cell Movement
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Cells, Cultured
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Chemokine CXCL12
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pharmacology
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Genetic Vectors
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genetics
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Humans
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Mesenchymal Stromal Cells
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cytology
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metabolism
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Receptors, CXCR4
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biosynthesis
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genetics
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Retroviridae
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genetics
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metabolism
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Transduction, Genetic
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Transfection
4.SDF-1/CXCR4 axis modulates bone marrow mesenchymal stem cell apoptosis, migration and cytokine secretion.
Xiaolei LIU ; Biyan DUAN ; Zhaokang CHENG ; Xiaohua JIA ; Lina MAO ; Hao FU ; Yongzhe CHE ; Lailiang OU ; Lin LIU ; Deling KONG
Protein & Cell 2011;2(10):845-854
Bone marrow mesenchymal stem cells (MSCs) are considered as a promising cell source to treat the acute myocardial infarction. However, over 90% of the stem cells usually die in the first three days of transplantation. Survival potential, migration ability and paracrine capacity have been considered as the most important three factors for cell transplantation in the ischemic cardiac treatment. We hypothesized that stromal-derived factor-1 (SDF-1)/CXCR4 axis plays a critical role in the regulation of these processes. In this study, apoptosis was induced by exposure of MSCs to H(2)O(2) for 2 h. After re-oxygenation, the SDF-1 pretreated MSCs demonstrated a significant increase in survival and proliferation. SDF-1 pretreatment also enhanced the migration and increased the secretion of pro-survival and angiogenic cytokines including basic fibroblast growth factor and vascular endothelial growth factor. Western blot and RT-PCR demonstrated that SDF-1 pretreatment significantly activated the pro-survival Akt and Erk signaling pathways and up-regulated Bcl-2/Bax ratio. These protective effects were partially inhibited by AMD3100, an antagonist of CXCR4.We conclude that the SDF-1/CXCR4 axis is critical for MSC survival, migration and cytokine secretion.
Animals
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Apoptosis
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Apoptosis Regulatory Proteins
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genetics
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metabolism
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Bone Marrow Cells
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metabolism
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physiology
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Cell Hypoxia
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Cell Movement
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Chemokine CXCL12
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genetics
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pharmacology
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physiology
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Cytokines
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metabolism
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Gene Expression
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L-Lactate Dehydrogenase
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metabolism
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MAP Kinase Signaling System
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Male
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Mesenchymal Stem Cells
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metabolism
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physiology
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Proto-Oncogene Proteins c-akt
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metabolism
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Rats
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Rats, Sprague-Dawley
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Receptors, CXCR4
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metabolism