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Background and Objectives@#Adipose tissue is a source of mesenchymal stem cells, which have the potential to differentiate into various types of cells. Adipose-derived stem cells (ADSCs) are now recognized as an accessible, abundant, and reliable stem cells suitable for tissue engineering and regenerative medicine applications. However, few literatures gave a comprehensive report on the capacities of ADSCs harvested from different sites. Especially, the capacities of ADSCs from aged mice remained unclear. In this study, we investigated several main capacities of brown adipose derived stem cells (B-ADSCs) and white adipose derived stem cells (W-ADSCs) from both young and aged mice. @*Methods@#and Results: When isolated from young mice, B-ADSCs showed a stronger proliferation rate and higher osteogenic, adipogenic and myocardial differentiation ability than W-ADSCs. Carboxy fluorescein diacetate succinimidyl ester (CFSE) labeling test suggested no significant difference in immunosuppression capacity between B-ADSCs and W-ADSCs. Similarly, no difference between these two were found in several immune related molecules, such as programmed death-ligand 1 (PD-L1), intercellular cell adhesion molecule (ICAM-1), vascular cell adhesion molecule (VCAM-1), inducible nitric oxide synthase (iNOS), tumour necrosis factor-α (TNF-α), interleukin 10 (IL10), and suppressor of cytokine signaling 1 (socs1). When isolated from aged mice, B-ADSCs also showed a stronger proliferation rate and higher osteogenic, adipogenic and myocardial differentiation ability than W-ADSCs; however, it demonstrated an attenuated immunosuppression capacity compared to W-ADSCs. @*Conclusions@#In summary, our data showed that ADSCs’ characteristics were tissue source dependent and changed with age. It provided evidence for choosing the right tissue-specific ADSCs for clinical application and fundamental research.
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<p><b>OBJECTIVE</b>To investigate the effect of microRNA-20a(MiR-20a) on osteogenic differentiation of mouse C3H/10T1/2 cells and its regulatory mechanism.</p><p><b>METHODS</b>Osteogenic differentiation of C3H/10T1/2 cells were identified by ALP staining and qRT-PCR. MiR-20a mimics and CKIP-1 siRNA were transfected into C3H/10T1/2 cells respectively with lipo3000. The expression of osteoblast marker genes, miR-20a and CKIP-1 were quantitatively assessed by qRT-PCR.</p><p><b>RESULTS</b>miR-20a expression was up-regulated during osteoblast differentiation of C3H/10T1/2 cells. Overexpression of miR-20a promoted osteogenic differentiation. Furthermore, miR-20a inhibited the expression of bone formation negative regulator CKIP-1. Additionally, CKIP-1 knockdown promoted osteogenic differentiation.</p><p><b>CONCLUSION</b>MiR-20a promotes osteogenic differentiation of C3H/10T1/2 cells possibly through inhibiting the expression of CKIP-1.</p>
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<strong>Objective</strong> To determine the mRNA and protein levels of urokinase plasminogen activator receptors (uPAR) in bone marrow fluid and bone marrow tissue from multiple myeloma (MM) patients and assess association of uPAR level with prognosis of MM. <strong>Methods</strong> uPAR levels in bone marrow fluid of 22 MM patients at the stable and progressive stages and 18 iron deficiency anemia patients with normal bone marrow (control) were examined by ELISA. Furthermore, uPAR expression in bone marrow tissue was investigated by RT-PCR and Western blot, respectively. The distribution of uPAR in MM cells was examined using immunofluorescence staining. The pathological changes in different stages of MM patients were studied by HE staining. <strong>Results</strong> uPAR level in bone marrow fluid of MM patients (1.52±0.32 μg/ml) was found to be higher than that in the control group (0.98±0.15 μg/ml). Interestingly, uPAR protein (0.686±0.075 vs. 0.372±0.043, P<0.05) and mRNA (2.51±0.46 vs. 4.46±1.15, P<0.05) expression levels of MM patients at the progressive stage were significantly higher than those at the stable stage. The expression of uPAR in MM bone marrow was confirmed by immunofluorescence staining. Moreover, HE staining revealed a great increased number of nucleated cells and severe impairment of hematopoietic function in the bone marrow of patients with progressive-stage myeloma. <strong>Conclusion</strong> Our study reveals that uPAR expression is positively correlated with the development and progress of MM.
Subject(s)
Adult , Aged , Female , Humans , Male , Middle Aged , Bone Marrow , Chemistry , Disease Progression , Fluorescent Antibody Technique , Multiple Myeloma , Chemistry , Pathology , Receptors, Urokinase Plasminogen ActivatorABSTRACT
Objective To determine the mRNA and protein levels of urokinase plasminogen activator receptors (uPAR) in bone marrow fluid and bone marrow tissue from multiple myeloma (MM) patients and assess association of uPAR level with prognosis of MM. Methods uPAR levels in bone marrow fluid of 22 MM patients at the stable and progressive stages and 18 iron deficiency anemia patients with normal bone marrow (control) were examined by ELISA. Furthermore, uPAR expression in bone marrow tissue was investigated by RT-PCR and Western blot, respectively. The distribution of uPAR in MM cells was examined using immunofluorescence staining. The pathological changes in different stages of MM patients were studied by HE staining. Results uPAR level in bone marrow fluid of MM patients (1.52±0.32 μg/ml) was found to be higher than that in the control group (0.98±0.15 μg/ml). Interestingly, uPAR protein (0.686±0.075 vs. 0.372±0.043, P<0.05) and mRNA (2.51±0.46 vs. 4.46±1.15, P<0.05) expression levels of MM patients at the progressive stage were significantly higher than those at the stable stage. The expression of uPAR in MM bone marrow was confirmed by immunofluorescence staining. Moreover, HE staining revealed a great increased number of nucleated cells and severe impairment of hematopoietic function in the bone marrow of patients with progressive-stage myeloma. Conclusion Our study reveals that uPAR expression is positively correlated with the development and progress of MM.
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Foot-and-mouth disease virus (FMDV) is the aetiological angent of a highly contagious viral disease. The complete gene encoding the structural protein of FMDV (P1) was subcloned into expression vector pGEX-KG, resulting in the fusion expression plasmid pKG-P1. After transformed into E. coli BL21(DE3) and induced by IPTG, the results of SDS-PAGE showed that the GST-P1 fusion protein was expressed in high level. The molecular weight of the fusion protein wa 110kD and the expressed products were soluble. Western-blotting was performed to confirm that the expressed fusion protein could specifically react with antiserum against FMDV. The fusion proteins were further purified by GST purification kit and an indirect ELISA (P1-ELISA) based on the purified proteins was developed. Comparison between P1-ELISA and the standard indirect haemagglutinin assay showed the two methods had 87 per cent agreement by detecting 864 serum samples, indicating the purified P1 protein was specific as the antigen of indirect P1-ELISA.
Subject(s)
Capsid Proteins , Genetics , Allergy and Immunology , Escherichia coli , Genetics , Metabolism , Foot-and-Mouth Disease Virus , Genetics , Recombinant Fusion Proteins , Genetics , Allergy and ImmunologyABSTRACT
<p><b>OBJECTIVE</b>To compare the effect of Astragulus injection (AGI) and interleukin-2 (IL-2) in enhancing anti-tumor metastasis action of dendrite cells (DCs) based vaccine.</p><p><b>METHODS</b>C57BL/6 mice's myelogenic DCs were prepared and pre-sensitized by Mut1, a MHC class I-restricted tumor antigen polypeptide of Lewis lung cancer. Then the DCs were used to treat mice with metastatic lung cancer in combination with AGI or IL-2. Change of proportion of T-lymphocyte cell subsets in splenic cell was analyzed by flow cytometry, and the serum contents of IL-2 and IL-4 of the tumor bearing mice were detected by ELISA.</p><p><b>RESULTS</b>After being treated with tumor antigen polypeptide sensitized DCs plus AGI or IL-2, the tubercle of lung cancer decreased, proportion of subsets CD4+T and CD8+T in mice's splenic cell increased, and serum IL-2/IL-4 ratio also increased obviously. During the observed period, the tumor developing rate in the immune mice treated with DCs combined treatment, either with IL-2 or with AGI, was lower than that in mice treated with DCs alone.</p><p><b>CONCLUSION</b>Both AGI and IL-2 can enhance the anti-tumor metastasis action of DCs, effectively promote the immune response of tumor bearing host, therefore have obviously inhibitory effect on lung cancer metastasis in vivo. Their immune protective function in normal animals is even more evident.</p>