Paeoniflorin Promotes Angiogenesis in A Vascular Insufficiency Model of Zebrafish in vivo and in Human Umbilical Vein Endothelial Cells in vitro / 中国结合医学杂志

<p><b>OBJECTIVE</b>To investigate the pro-angiogenic effects of paeoniflorin (PF) in a vascular insufficiency model of zebrafish and in human umbilical vein endothelial cells (HUVECs).</p><p><b>METHODS</b>In vivo, the pro-angiogenic effects of PF were tested in a vascular insufficiency model in the Tg(fli-1:EGFP)y1 transgenic zebrafish. The 24 h post fertilization (hpf) embryos were pretreated with vascular endothelial growth factor (VEGF) receptor tyrosine kinase inhibitor II (VRI) for 3 h to establish the vascular insufficiency model and then post-treated with PF for 24 h. The formation of intersegmental vessels (ISVs) was observed with a fluorescence microscope. The mRNA expression of fms-like tyrosine kinase-1 (flt-1), kinase insert domain receptor (kdr), kinase insert domain receptor like (kdrl) and von Willebrand factor (vWF) were analyzed by real-time polymerase chain reaction (PCR). In vitro, the pro-angiogenic effects of PF were observed in HUVECs in which cell proliferation, migration and tube formation were assessed.</p><p><b>RESULTS</b>PF (6.25-100 μmol/L) could rescue VRI-induced blood vessel loss in zebrafish and PF (25-100 μmol/L), thereby restoring the mRNA expressions of flt-1, kdr, kdrl and vWF, which were down-regulated by VRI treatment. In addition, PF (0.001-0.03 μmol/L) could promote the proliferation of HUVECs while PF stimulated HUVECs migration at 1.0-10 μmol/L and tube formation at 0.3 μmol/L.</p><p><b>CONCLUSION</b>PF could promote angiogenesis in a vascular insufficiency model of zebrafish in vivo and in HUVECs in vitro.</p>

Preparation of egg yolk antibodies against human isomaltase and determination of their biological activities / 医学研究生学报

Objective Antibody drugs are one of the hot topics in biomedical research.This study aims to develop egg yolk antibodies (IgYs) against human isomaltase and determine their biological activities.Methods The purified recombinant isomaltase protein was used as an antigen to immunize egg-laying hens in combination with complete Freund adjuvant (CFA).Anti-isomahase IgYs were extracted by water dilution-sodium sulfate extraction assay and further analyzed for their purity,specificity,titer and stability by SDS-PAGE,Western blot and ELISA respectively,and their inhibitory effect on human alpha-glycosidase enzymes was evaluated by the PNPG method.Results Anti-isomaltase IgYs were obtained,with a titer of 1 ∶ 12800,capable of specifically binding human isomaltase,and with a good thermal stability,acid/alkali stability and pepsin resistance.Conclusion Anti-human isomaltase IgYs were successfully prepared,which may provide an experimental ground for further investigation of oral antihyperglycemic agents for type Ⅱ diabetes mellitus.