Objective Study on the distribution of auto‐antibodies against platelet in adults and children with primary immune thrombocytopenia(ITP) .Methods Platelet auto‐antibodies were detected in 83 ITP patients and 58 non‐ITP patients by enzyme linked immunosorbent assay (PAKAUTO kit) .Anti‐GP Ⅱ b/Ⅲ a ,GP Ⅰ b/Ⅸ and GP Ⅰ a/Ⅱ a auto‐antibodies were analyzed for 46 a‐dults and 37 children with ITP .Results The positive rate of autoantibody was 66 .27% in ITP patients ,which was much higher than that of non‐ITP patients ,the difference was significant(P < 0 .05) .As for ITP patients ,female ITP patients in adult group were 28 ,while in children group were 24 ,the difference was not significant(P > 0 .05) ,but the morbidity of ITP in females was higher than that of males in both of the two groups ,the difference was significant(P< 0 .05) .There was no significant difference on the distribution of platelet auto‐antibodies in adult and children group(P> 0 .05) ,the main of them was resistance against GP Ⅱ b/Ⅲ a and GP Ⅰ b/Ⅸ antibodies .Conclusion The detection of platelet auto‐antibodies could distinguish ITP and non‐ITP patients ,and GP Ⅰ a/Ⅱ a has an important reference value about therapy .

Objective To organize the 14th ISBT Platelet Immunology Workshop and Cooperative Research Project,and proficiency evaluation on the techniques and quality level of platelet alloantiboby analysis will be taken effect for the 42 platelet immunology laboratories around the world. Methods Organized and confirmed by Nanning Institute of Transfusion Medicine,9 quality control samples contained the Human Platelet Antigen (HPA) specificity antibody have been supplied to all the participated laboratories in this project. The participants could use the In-House technology or/and market kits to test these quality control samples. The forms and sheets have been provided for recording of results. Results There were 36 laboratories from 23 countries participated in this Cooperative Research Project,and 35 laboratories have reported their results. The appraised consistent rate of the 9 quality control samples ranges from 20% to 97.14%,the HPA antibody specificities have been showed as anti HPA-1a,anti HPA-1b,anti HPA-3a,anti HPA-3a,anti HPA-3b,anti HPA-5b,anti HPA-5a,anti GPIV and anti HPA-5b+15b,respectively. Among the appraised consistent rate,the anti HPA-3b were the lowest and the anti HPA-5b and anti HPA-5a was the highest. Over 10 technologies of platelet alloantiboby analysis were used by the laboratories. Conclusion This international cooperative research project has successfully made the proficiency evaluation and report on the techniques and quality level of platelet alloantiboby analysis for the 35 international platelet immunology laboratories.

Objective To establish the platelet antigen panel cells and to apply them in the detection and identification of platelet alloantibody.Methods Human platelet antigen(HPA)1-16 genotyping from 1 500 un-related blood donors in Nanning area were performed by the polymerase chain reaction-sequence specific primers(PCR-SSP)technique,platelet antigen cells with O blood type were chosen to establish the panel cells of platelet antigen.The phenotype of the panel cells were verified by the reference sera from the 14th platelet immunology workshop of the International Society of Blood Transfusion(ISBT).And then these panel cells were used in clinic to detect the platelet alloantibody and the samples from the 14th and 15th platelet immunology workshop of ISBT.Re-sults Six platelet cells with consistent phenotype and genotypes and covering the HPA 1-5 and 1 5 systems were selected to estab-lish the platelet panel cells and successfully applied them in the clinical and scientific sample detection and identification.Conclusion Platelet antigen panel cells are established successfully,which provides the experimental basis for the diagnosis and research of platelet allogenic abnormal immunity diseases.

Objective To investigate the distribution situation of ABO and Rh blood groups among Zhuang population in Nanning area to guide the scientific voluntary blood donor recruitment and rational storage of blood in this area.Methods 2 052 blood samples from Zhuang population in Nanning area were performed the identification of ABO and Rh blood groups.Whether the gene distributions conforming to the Hardy-Weinberg equilibrium principle was assessed by using the Chi-square test.The data was compared with the distribution characteristics in other areas or other minorities.Results The distribution characteristics of ABO blood groups in Nanning area were O > B> A > AB.Blood group O was highest (46.78%)and blood group AB was lowest (4.34%).The gene frequencies of A,B and O were 0.131 9,0.181 5 and 0.686 6,respectively.The blood groups distribution con-formed to the Hardy-Weinberg equilibrium principle;the RhD positive proportion(RhD+ )in Zhuang population was 99.90%,while the RhD negative (RhD- )proportion was 0.10%,which was lower than that in Han population.No phenotypes of ccdE,CcdE, CCdee,CCdE,CCDEE and ccDee were observed in this study.Conclusion The distribution of the ABO and Rh blood groups among Zhuang population in Nanning still maintains their own exclusive characteristics.The extremely low proportion of RhD- will bring about serious difficulties to the patients with long term blood transfusion.

OBJECTIVETo explore the molecular basis for a CD36 deficiency individual and distribution of CD36 gene mutation in Guangxi population.

METHODSA female individual was studied. CD36 phenotype was detected by monoclonal antibody immobilization of platelet antigens assay (MAIPA) and flow cytometry (FCM). The coding regions of the CD36 gene were sequenced. A DNA-based polymerase chain reaction-sequence specific primer (PCR-SSP) assay was used to verify the identified mutation. Cell lines expressing the mutant and wild-type CD36[CD36(MT) and CD36(WT)] were established, with the expression of CD36 determined by Western blotting. The distribution of CD36 gene mutation was investigated among 1010 unrelated individuals with the PCR-SSP assay.

RESULTSBoth MAIPA and FCM assays showed that the patient had type II CD36 deficiency. DNA sequencing showed that she has carried a heterozygous mutation T538C (Trp180Arg) in the exon 6 of CD36. Sequencing of cDNA clone confirmed that there was a nucleotide substitution at position 538 (538T>C). Western blotting also confirmed that the CD36 did not express on the CD36(MT) cell line that expressed the 538C mutant, but did express on the CD36(WT) cell line. The novel CD36 mutation T538C was further verified with 100% concordance of genotyping results by DNA-based PCR-SSP assay and 1010 unrelated individuals. No CD36 538C allele was detected among the 1010 individuals.

CONCLUSIONThis study has identified a novel CD36 mutation T538C(Trp180Arg)(GenBank: HM217022.1), and established a genotyping method for the novel sequence-specific primer PCR. The novel mutation is rare in Guangxi and can cause type II CD36 deficiency.

Base Sequence ; Blood Platelet Disorders ; genetics ; Blood Platelets ; cytology ; metabolism ; Blotting, Western ; CD36 Antigens ; genetics ; metabolism ; Cells, Cultured ; DNA Mutational Analysis ; DNA Primers ; genetics ; Exons ; genetics ; Female ; Flow Cytometry ; Fluorescent Antibody Technique ; Genetic Diseases, Inborn ; genetics ; Genotype ; Genotyping Techniques ; methods ; Humans ; Middle Aged ; Monocytes ; cytology ; metabolism ; Mutation, Missense ; Polymerase Chain Reaction ; methods