ABSTRACT
Objective To compare the transfection efficiency of cationic polyethylenimine(PEI)with Lipofectamine 2000TM by using the plasmid DNA encoding vascular endothelial cell growth factor (VEGF165 )gene in human embryonic kidney cell line 293T.Methods PEI of different N/P ratio and Lipofectamine 2000TM were used to deliver the vector containing VEGF165 to 293T cells,respectively.Green fluorescent protein(GFP)gene was inserted into the vector as a report gene.Evaluation of cytoactive was performed by CCK-8 assay 24 h after transfection.The cells were observed by fluorescent microscope and the presence of VEGF165 in cell supernatant was detected by ELISA 48 h after transfection.The transfection efficiency was calculated and com-pared.Results Similar cytoactive and best transfection efficiency could be obtained when N/P ratio was 9,the transfection efficien-cy was around 70%.Furthermore,the presence of VEGF165 increased significantly after transfection(P <0.05),but there was no significant difference between the two groups in which different transfection methods were adopted.Conclusion PEI as a novel oli-gofectamine reagent could mediate more efficient transfection compared with lipofectamine.It also has low cell-toxicity and low price and could be an ideal vector in gene delivery technology.
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Objective To analyze the relevance between the point-of-care testing (POCT) and routine test in BNP testing .Meth-ods The whole blood samples or plasma samples from 40 inpatients were detected brain natriuretic peptide (BNP) by the Alere Triage? MeterPro fluorescence immunoassay analyzer (POCT ) or the Beckman Coulter Access ?2 chemiluminescence analyzer (routine test) ,respectively .The acquired data were subjected to comparative analysis according to the CLCS EP 9-A2 .Results The linear regression of the BNP content in the blood samples detected by POCT and the routine test was good ,the correlation coeffi-cient(r) was 0 .999 7 .Conclusion POCT and the routine test have good correlation in BNP testing .POCT for BNP testing has higher reliability and is applicable for clinical detection .
ABSTRACT
<p><b>OBJECTIVE</b>To study the damage of mitochondrial function of sperm induced by reactive oxygen species (ROS), and the protection of melatonin (MLT) against the damage.</p><p><b>METHODS</b>Spermatozoa of normal physiological function selected from semen samples by Percoll gradient centrifugation technique were used as normal sperm models in the present study. Reactive oxygen species were generated by hypoxanthine xanthine oxidase system, and in the presence (or absence) of MLT (6 mmol/L), incubated with normal sperm models for 30 and 60 minutes. After incubation, the activity of succinate dehydrogenase (SDH) in mitochondria of spermatozoa was assessed by histochemical method, and spermatozoa were labeled with specific fluorescent probe of Rhodamine 123 to measure mitochondrial membrane potential (MMP) by flow cytometry.</p><p><b>RESULTS</b>After normal spermatozoa were incubated with ROS, MMP of spermatozoa significantly decreased, and the activity of SDH almost decreased to zero. However, MLT had effect on reducing the damage of the mitochondrial function of sperm induced by ROS.</p><p><b>CONCLUSION</b>ROS can damage the mitochondrial function of sperm by affecting MMP of spermatozoa and the activity of SDH. MLT can protect sperm mitochondria from the damage induced by ROS through its effective antioxidative potential.</p>
Subject(s)
Humans , Male , Flow Cytometry , Melatonin , Pharmacology , Membrane Potentials , Mitochondria , Physiology , Reactive Oxygen Species , Pharmacology , Spermatozoa , Physiology , Succinate Dehydrogenase , MetabolismABSTRACT
Increased generation of ROS causes the lipid oxidation of the membrane of spermatozoa, but antioxidant vitamins play an important role in reproduction and help clear away ROS and protect the sperm membrane from lipid oxidation. This review focused on the effect of antioxidant vitamins on male reproduction and in the treatment of male infertility.