ABSTRACT
Drug-induced immune hemolytic anemia (DIIHA) is a rare cytopenia caused by damage to RBCs by drug-induced antibodies or non-immune protein adsorption (NIPA). The drugs associated with DIIHA and the mechanistic hypotheses that are thought to be involved have been controversial, with complex serological tests often required by specialized Immune Hematology laboratories for diagnosis. It is necessary to know the clinical manifestation and laboratory diagnosis of DIIHA in order to distinguish the immuno-hematological abnormality caused by drugs from other causes. How to improve the diagnostic ability of DIIHA and establish a scientific and reasonable idea of DIIHA serological examination is urgent to help clinical diagnosis and correct treatment.
ABSTRACT
【Objective】 To investigate the effect of ssDNA aptamer of RhD blood group antigen on erythrocyte toxicity. 【Methods】 Two full-length ssDNA aptamers(82 bp) of RhD blood group antigen were obtained by gene synthesis.Five samples of whole blood with EDTA anticoagulant were collected to prepare red blood cell suspensions (4×107/mL), which were split into 10 tubes(100 μL/tube), corresponding to 5 experimental groups and 5 controls.Two monospecific full-length ssDNA sequences (100 pmol/μL, 5μL each) were added into the experimental group, while the same amount of normal saline into the control.After treatment, the experimental group and the control were incubated for 60 min at 37℃.After washing, they were suspended in LISS solution and stored at 4℃.The experimental group and the control were set according to different time point during storage (1 h, 1 d, 3 d, 10 d and 17 d), with 5 tubes in each group.For erythrocytes in LISS suspension at different storage time, Annexin V labeled with FITC was used as a probe to label the phosphatidylserine (PS) content and Fluo-4 to label Ca2+ .The eversion of PS and the change of Ca2+ concentration in red blood cells in LISS suspensions were determined by flow cytometry. 【Results】 After incubation, all groups were examined under the light microscope.No agglutination occurred in the experimental group, while agglutination occurred in the control.Flow cytometry showed that the number of Annexin V-FITC staining cells of suspended erythrocytes at the same storage time-point was similar between the experimental group and the control, with no significant differences.In the experimental group, apoptosis rate of Annexin V cells at 10-day storage(6.06±1.38) was significantly higher than that at 1-hour storage(P<0.05), so as at 17-day storage(7.77±1.23) than 1-hour, 1-day and 3-day storage(P<0.05). The apoptosis rate of Fluo-4 AM cell in suspended RBCs at the same storage time-point was similar between the two groups(P>0.05). In the experimental group, the apoptosis rate of Fluo-4 AM cell at the 3-day, 10-day and 17-day storage was 20.84±4.16, 22.35±3.37 and 27.06±2.81, respectively(P<0.05). 【Conclusion】 ssDNA aptamer was not found to have any cytotoxic effects on red blood cells, and RhD ssDNA aptamer may be used as a material for the detection and preparation of universal blood.
ABSTRACT
Objective To analyze the change of phosphotidyl serine(PS) in apheresis platelets(plts) during storage process,and to evaluate the quality of the plts during the storage period.Methods The apoptosis ligand Fas(CD95) was used to induce plts as apoptosis model,and by which the method to detect plts PS by flow cytometry(FCM) was established.Subsequently the resting PS expressive rate and the induced PS expressive rate of 30 apheresis plts,which induced by CD95,were determined using the FCM method established in storage process.Results At the 1st,5th,7th,8th day of storage process,the PS expression of plts kept on increasing.The PS expressive rate of plts were 2.03%?0.91%,3.26%?1.86%,8.98%?4.60%,26.68%?21.28%,and 38.85%?26.43%,respectively,and there were significant differences among groups respectively(P0.05).Conclusion The PS expression of apheresis plts increase significantly with the prolongation of storage time;Fresh plts has the ability of anti-apoptosis;Apheresis plts could keep normal function until storage 7 d.
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Objective To gain purified platelets for further research of the immune function of platelet.Methods Platelets were purified by positive(MS) and negative(LD) magnetic cell sorting(MACS) separation.The percentage of activated platelets was detected by flow cytometry and nucleated cell clearance was evaluated by white cell count.Results The percentage of activated platelets before separation was(2.39?1.10)%,and increased to(2.56?1.08)% and(16.76?4.04)%,respectively,after MS and LD MACS.The clearances of nucleated cells after MACS MS and LD were(98.44?0.24)% and(98.47?0.18)%,respectively.The recovery rate of purified platelet after MS and LD was(76.50?1.49)% and(79.20?2.61)%.Conclusion MACS LD method was more suitable for the platelet purification.
ABSTRACT
Objective:To study on significance of ABO genotyping.Methods:To use the methods of polymerase chain reaction sequence specific primers(PCR SSP) for genotyping of ABO blood group and observation ABO gene polymorphism as well as identification of doubt sample.Results:The reliability of the genotyping for ABO blood group was proved by testing DNA samples previously known genotypes.The results of genotyping for 104 healthy and unrelated HAN individuals were correspond with serological phenotypes.The method of ABO genotyping was applied for clinical pre transfusion ABO typing,pre delivery fetal ABO typing,parenting test and subgrouptyping.Conclusion:The method of ABO genotyping may correct typing for doubt sample of ABO serological typing.