1.Establishment of mannitol-induced infusion phlebitis animal model
Linsheng SHEN ; Ping WANG ; Dengke XIAO ; Ruyan ZHANG ; Yuanyuan MIAO ; Yulu GUO
Chinese Journal of Practical Nursing 2014;30(23):9-11
Objective To explore the reasonable experimental parameters on establishment of rabbit model of infusion phlebitis induced by mannitol.Methods New Zealand rabbits were injected with 20% mannitol,then pathological lesion of rabbit auricular vein induced by different infusion velocity,different sampling time and sites were observed under microscope with vascular injury,inflammatory cell infiltration,frequency of thrombokinesis as indexes.Results The three indexes were the highest and the most obvious characteristics of infusion phlebitis were noted at the following experimental conditions:5.0 ml/min (infusion velocity),sampling time at 24h after administration and sampling site at 1cm region in front of the catheter tip.Conclusions Rabbit model of infusion phlebitis induced by mannitol can be set up more stable by using these parameters.
2.Study on SCARB1 mediated coagulation dysregulation in recipients after liver xenotransplantation
Xiao LI ; Hongchen JI ; Kefeng DOU ; Dengke PAN ; Hui CHEN ; Liang ZHOU ; Kaishan TAO ; Zhengcai LIU
Organ Transplantation 2017;8(2):115-120
To investigate the changes in the expression levels of scavenger receptor class B member 1 (SCARB 1) in the liver tissues before and after liver xenotransplantation and analyze the relationship between the variations in the SCARB1 expression and coagulation regulating dysfunction in the recipients.Methods The Wuzhishan miniature pig with α-1,3-galactosyltransferase gene-knockout(GTKO) was utilized as the donor and Macaca thibetana was chosen as the recipient.Heterotopic auxiliary liver xenotransplantation models were established.The liver tissue specimen was collected before and after liver xenotransplantation.Primary hepatocytes were extracted from the pig using collagenase digestion method.Human peripheral blood mononuclear cells were obtained by immunomagnetic bead sorting.These two types of cells were co-cultured and supplemented with human plasma to establish cell models with coagulation regulating dysfunction following liver xenotransplantation.Quantitative real-time reverse transcription polymerase chain reaction (qRT-PCR) and Western blot were performed to quantitatively measure and statistically compared the expression levels of messenger ribonucleic acid (mRNA) and protein of SCARB1 in the tissue and cell samples.At the cellular level,the expression of SCARB 1 was interfered by lentiviral vector.The coagulation time was detected to validate the effect upon coagulation function.Results The expression levels of SCARB1 mRNA and protein were significantly down-regulated after liver xenotransplantation (both P<0.05).In the cell models,the expression levels of SCARB1 mRNA and protein in the porcine hepatocytes co-cultured with human monocytes were significantly down-regulated compared with those in porcine hepatocytes without intervention (both P<0.05).Compared with the non-intervention group,the coagulation time was significantly prolonged after the expression of SCARB1 was interfered by lentiviral vector (P<0.05).Conclusions The down-regulated expression of SCARB1 in the liver graft is one of the main causes of mediating coagulation regulating dysfunction.Intervention of SCARB1 expression contributes to resolve the coagulation regulating dysfunction in the recipients after liver xenotransplantation.