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Objective To investigate the effects of training of Bobath position transfer technique for nurses.Methods Thirty nurses were recruited in the study using convenience sampling and received training of Bobath transfer technique.Five variables were evaluated before and after training:transfer skills,transfer intensity,sense of comfort,and sense of comfort and sense of security from simulated patients.Results Nurses' transfer skills and sense of comfort were higher after training (P＜0.05).Transfer intensity was lower than before (P＜0.05).Sense of comfort and sense of security from simulated patients were improved after the training (P＜0.05).Conclusion Training of Bobath transfer technique can improve nurses' transfer skills and promote sense of comfort and sense of security from simulated patients.
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<p><b>OBJECTIVE</b>To explore the pathogenesis of a child with growth retardation, liver damage and congenital heart disease.</p><p><b>METHODS</b>G-banded chromosomal karyotyping, high-throughput next-generation sequencing (HT-NGS)and fluorescence in situ hybridization(FISH) were used to characterize the structural chromosomal aberration.</p><p><b>RESULTS</b>The child was found to have a karyotype of 46, XX, t(1;2) (q25;q21), t(7;20) (q21;p13). HT-NGS has detected a microdeletion at 2q21.3 and 7q21.11, respectively, which were verified by FISH.</p><p><b>CONCLUSION</b>Combined cytogenetic and molecular analysis can detect chromosome micrdeletions more precisely. The abnormalities of the child may be attributed to heterozygous deletion of ZEB2, ABCB4 and SEMA3A genes.</p>
Subject(s)
Female , Humans , Infant , Chromosome Aberrations , Chromosome Banding , Methods , Heart Defects, Congenital , Genetics , Intellectual Disability , Genetics , Karyotyping , Methods , Liver Diseases , GeneticsABSTRACT
<p><b>OBJECTIVE</b>To observe the effect of rosiglitazone on the content of cholesterol and expressions of Acy-coenzyme A: cholesterol acyltransferase 1 (ACAT-1) and scavenger receptor class B type I (SR-BI) in RAW264.7 macrophage-derived foam cells and explore the anti-atherosclerotic mechanism of rosiglitazone.</p><p><b>METHODS</b>RAW264.7 macrophages were incubated with oxidized low-density lipoproteins (ox-LDL) or with both ox-LDL and rosiglitazone (5, 10, or 20 µmol/L). Oil red O staining was used to observe the formation of foam cells, and cholesterol oxidase was used to determine the content of cellular cholesterol contents. Western blotting was used observe the expressions of ACAT-1 and SR-BI in RAW264.7 foam cells.</p><p><b>RESULTS</b>Compared with the control cells, RAW264.7 macrophage-derived foam cells showed significantly increased contents of total cholesterol and free cholesterol (P<0.01) and ACAT-1 expressions (P<0.05) with mildly increased SR-BI expression (P>0.05). Rosiglitazone treatments significantly lowered the contents of total cholesterol and free cholesterol (P<0.05), decreased the expression of ACAT-1 (P<0.05), and increased SR-BI expression (P<0.05) in the foam cells in a dose-dependent manner.</p><p><b>CONCLUSION</b>Rosiglitazone can decrease the contents of total and free cholesterol, down-regulate ACAT-1 expression and up-regulate SR-BI expression in the foam cells produce the anti-atherosclerotic effect.</p>
Subject(s)
Humans , Acetyl-CoA C-Acetyltransferase , Metabolism , Cell Line , Cholesterol , Metabolism , Foam Cells , Cell Biology , Metabolism , Scavenger Receptors, Class B , Metabolism , Thiazolidinediones , PharmacologyABSTRACT
Objective To clone ORF of DcR3 gene and insert it into eukaryotic expression vector and express it in COS-7 cells. Methods Encoding sequence of human DcR3 gene was cloned by PCR and sequenced. The sequenced ORF was cloned into eukaryotic expression vector pAAV-IRES-hrGFP to construct recombinant plasmid. COS-7 cells were transfected with recombinant plasmid by lipofectamine2000. Expression of recombinant DcR3 gene was verified by Western blotting and confocal microscopy. Results A 1 000-bp gene segment was obtained by PCR and inserted into pAAV-IRES-hrGFP to construct recombinant plasmid. The gene segment was proved to be encoding sequence of human DcR3 gene by sequencing. DcR3 expression in COS-7 cells was verified by Western blotting and confocal microscopy. Conclusion DcR3 gene was successfully cloned and expressed in COS-7 cells.
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Objective To clone the full length cDNA of human LIGHT and to construct the recombinant eukaryotic expression plasmid pCI neo LIGHT for the stable expression on 293T cells. Methods Human LIGHT cDNA was cloned from a normal human activated T cell cDNA library phAD.CAD by PCR. After sequencing, LIGHT cDNA was inserted into plasmid pCI neo for the construction of the eukaryotic expression. The recombinant was transfected into 293T cells by electroporation. The expression of LIGHT on the surface of 293T cells was detected by flow cytometry after screening with G418. Results Sequencing confirmed that ORF of LIGHT gene was intact and right. Restrictive enzyme digestion proved that LIGHT gene was inserted into the recombinant plasmid of LIGHT pCI neo correctly. FACS analysis revealed that about 78.69% 293T cells expressed LIGHT protein on the cell surfaces at 3 months after screening with G418. Conclusion LIGHT gene has been cloned successfully, and a 293T cell line expressing LIGHT protein on its membrane surface has been obtained.
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Objective To construct eukaryotic expression plasmid of hCD137 Fc gene and express hCD137 Fc fusion protein with high biological activity. Methods PCR technique was employed to clone the human CD137 cDNA from a normal human activated T cells cDNA library, and then clone its extramembrane encoding region. The extramembrane sequence together with human IgG1 Fc cDNA were inserted into the eukaryotic expression plasmid pcDNA3. CD137 Fc gene was expressed transiently in 293T cells, and then CD137 Fc protein was purified by recombinated protein A affinity chromatography column. At last, the MW, purity and antigenicity of CD137 Fc were identified by sandwich ELISA, SDS PAGE and Western blotting, respectively. Results The ORF of CD137 Fc gene was coincident with what we expected. ELISA and SDS PAGE confirmed protein expression in 293T cells. Western blotting proved the antigenicity of the purified CD137 Fc protein. Conclusion We obtained a purified recombinated CD137 Fc protein with biological activity and the expected MW. This lays the foundation for further studies of CD137 such as its role in immune homeostasis and other biological functions.
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Objective To explore a new simpler method for the preparation of recombinant alpha virus as a novel vaccine at the DNA level. Methods Plasmids expressing ? gal protein and helper plasmids were transfected into BHK cells. Virus in culture supernatant of the transfected BHK cells were collected and purified and used to infect BHK cells in vitro to identify the expression of target gene and the titre of the recombinant virus. Results Recombinant virus with high titre, prepared by this method, could be expressed well in mammalian cells in vitro . Conclusion High titre recombinant alpha virus can be produced at the DNA level and this method can be applied for vaccine preparation and gene therapy.
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Objective:To express the GPI-anchored CD55 and recombinant transmembrane form CD55 molecules on mouse fibroblasts cell line NIH3T3, and compare their inhibitory function of complement lysis.Methods:In previous study,had constructed the transmembrane form CD55 (CD55-TM) cDNA by linking the extracellular portion of CD55 to the transmembrane and cytoplasmic domains of MCP, and then subcloned into retroviral vector pLXSN. In this experiment,had transfected recombinant CD55-TM and GPI anchored CD55 into PA317 packaging cell to generate stable virus-producing cell lines. And then, mouse fibroblasts cell line NIH3T3 was infected with the virus containing CD55, recombinant CD55-TM or pLXSN alone. The expression of these molecules on NIH3T3 cells was detected by FACS analysis. Complement killing assay was carried out using MTT colorimetric method.Results:FACS analysis showed that both CD55 and its TM version were expressed stably on NIH3T3 cells. Both kinds of CD55 molecules can efficiently protect the NIH3T3 cells from complement mediated lysis and there is no significant difference between them. Conclusion:These data showed that both GPI-anchored CD55 and its TM version can normally expressed on NIH3T3 cells and both can protect the NIH3T3 cells from human complement mediated lysis. These results confirmed the feasibility of TM CD55 based gene therapy could be used for PNH, and also provided an excellent model for the study of signal transduction mechanisms mediated by GPI-anchored protein.