Objective To observe the difference between the bone marrow and peripheral blood engraftment evidence after allogeneic haemopoietic stem cell transplantation(Allo-HSCT) using PCR-STR. Methods DNA from peripheral blood or bone marrow of donors and recipients in different phases were extracted,and 16 STR loci with high polymorphism were amplified by PCR.Separation of the PCR products and fluorescence detection were performed by ABIprism 3100 Genetic Analyzer with capillary electrophoresis.Results The 16 patients included in the study had different levels of engraftment.Twelve patients displayed complete chimerism,while 5 patients showed mixed chimerism.One patient was keeping continuance of remission.The decrease of donor DNA amounts in mixed chimerism was earlier in bone marrow than that in peripheral blood(P

Objective To study the polymorphism of human platelet antigens among unrelated Guangzhou blood donors with PCR-SSP,in order to provide basic data for population studies and clinical transfusion practice.Methods Blood samples from 706 unrelated blood donors in Guangzhou were genotyped for each of the HPA1—6,15 systems by PCR-SSP.Gene frequencies and genotype frequencies were analyzed by statistical methods.Results HPA-3 and-15 had the greatest heterozygosity with a gene frequency of 0.2918,0.4830,0.2252 for HPA-3a/a,HPA-3a/b,HPA-3b/b,and 0.2691,0.5170,0.2139 for HPA-15a/a,HPA-15a/b,HPA-15b/b.The a/a homozygosity was predominant in HPA-1,-2,-4,-5,with a frequency ranged from 0.9583 to 0.9993,while HPA b/b was not found among them.The frequency of HPA-lb and HPA-4b was very low,which was 0.0028 and 0.0007,respectively.In our study,HPA-1 frequency was significantly different from that of the north Chinese,English,and American Indian(P

Objective To establish eight human platelet antigen systems and HLA-Ⅰ antigen donor bank,and to determine the gene frequencies of human platelet antigen(HPA) and HLA-Ⅰin Guangzhou area.Methods A total of 805 blood samples from Chinese Han voluntary platelet donors were included in this study.PCR-SSP was used to detect single-nucleotide polymorphism in HPA systems.Luminex-SSO was used to detect the HLA-Ⅰantigens.Results The distribution of HPA 1,2,3,4,5,6,9,15 was in Hardy-Weiberg equilibrium among study subjects.Allele frequencies of 0.998 and 0.002 were observed for HPA 1a and 1b,0.952 and 0.048 for HPA 2a and 2b,0.553 and 0.447 for HPA 3a and 3b,0.999 and 0.001 for HPA 4a and 4b,0.976 and 0.024 for HPA 5a and 5b,0.982 and 0.018 for HPA 6a and 6b,1 and 0 for HPA 9a and 9b,0.518 and 0.481 for HPA 15a and 15b.The high frequency HLA-Ⅰ alleles were A*02,0.286;A*24,0.162;A*11,0.323;B*46,0.147;B*75,0.100;C*01,0.177;C*03,0.289;C*07,0.179.Conclusions This study confirmed the ethnic and territorial difference of HPA and HLA-Ⅰ.The establishment of HPA and HLA-Ⅰ matched plateletpheresis donor registry is helpful in the improvement in platelet transfusion.

AIM: To establish human lymphocyte cell line secreting monoclonal antibody against Rhesus(D) antigen and analyse the nucleotide and deduced amino acid sequences of a human monoclonal anti-D Fab fragment. METHODS: By using PCR method, the cDNA of human anti-(Rhesus D) antibody(lgM ?)Fab fragment was amplified from an Epstein- Barr-virus-transformed cell line. Cloning and subsequent sequence analysis of the Fab fragment was performed. The deduced amino acid sequence was compared and analysed with previously published sequences. RESULTS : A band of approximate 700 and 650 base pairs was amplified using lgM heavy chain primers and ? light chain primers, respectively. Sequence analysis indicated that the deduced amino acid sequences was in agreement with the characterization of the amino acid present in the human lg Fab fragment. CONCLUSION: The cloning and sequencing of a human anti-Rhesus (D) antibody Fab fragment cDNA will make benefits for production of recombinant anti-Rhesus (D) antibody and prevention of Rh haemolytic disease in the newborn.

The severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is the pathogen that causes novel coronavirus pneumonia. The SARS-CoV-2 mainly transmits through respiratory tract. However, RNA of this virus can be detected in blood samples of some infected cases. This paper herein reviewed the risk of transfusion transmission of SARS-CoV-2 and relevant preventive measures. The impact of SARS-CoV-2 endemic on blood supply and the corresponding strategy were also discussed in this article.

Objective Study on producing human monoclonal antibodies against Rhesus (D) antigen that was suitable for use as blood group typing reagent. Methods B lymphocytes from a Rh negative woman, which can produce anti D antibodies were transformed by Epstein Barr virus(EBV). Antibody secreting cells were enriched by RhD + group O erythrocytes and cloned by limited dilute method. By using one step emzymatic method on microplates, one thousand normal blood donors with a common Rh phenotype were tested with the supernatant of cell culture medium as well as a polyclonal human anti D and a commercial monoclonal anti D serum. Results Three human B lymphocyte lines secreting monoclonal antibodies to Rh (D) were established. One of them produced lgM antibody. The titer of the monoclonal antibodies was 64～128. Study on 1000 blood donors, the results did not show any discrepancy among the three different anti Rh(D) serum. Conclusion These monoclonal antibodies against D antigen could be used in Rh(D) typing.

AIM: To investigate the expression of B lymphocyte stimulator(BLyS) and CD38 molecules on peripheral blood lymphocytes of patients with systemic lupus erythematosus(SLE). METHODS: Twenty-two patients with SLE and fourteen healthy subjects entered the study. Isolated peripheral blood lymphocyte were stained for the lymphocyte surface markers BLyS, CD19, and CD38, and then was measured by flow cytometry(FACS). RESULTS: BLyS + lymphocytes, CD19 + lymphocytes, and CD19 +CD38 + lymphocytes were increased significantly in patients with SLE( P

AIM: To amplify from leader peptide region an d obtain human monoclonal anti-D variable region gene with high specificity and affinity, and analyze the nucleotide and deduced amino acid sequences.ME THODS: The total RNA was extracted from an Epstein-Barr-virus-transforme d cell line secreting monoclonal anti- (rhesus D) antibody. The leader region pri mers containing a ribosome recognition site were designed. By using PCR method, the cDNA of human anti-(rhesus D) antibody (IgM ?) variable region gene was amp lified. Cloning and subsequent sequence analysis of the variable region gene was performed. The deduced amino acid sequence was also compared and analyzed with previ ously published sequences.RESULTS: A band of approximate 440 and 410 base pairs were amplified using heavy chain primers and light chain primer s, respectively. Sequence analysis indicated that the deduced amino acid sequenc e w as in agreement with the characterization of the amino acid present in the human Ig variable region. CONCLUSION: The cloning and sequencing of a human anti- (Rhesus D) antibody variable region cDNA will make benefits for pro duction of recombinant anti-(Rhesus D) antibody and prevention of Rh haemolytic disease in newborns.

AIM: To clone and express a human monoclonal anti-D Fab fragment in E. coli and make benefits for the expression of the whole immunoglobulin molecules of anti-D. METHODS: The gene of anti-D Fab fragment was cloned into the phagemid vector pComb3. After analyzing by PCR and restriction site analysis, the recombinant was expressed in E. coli and the expressed protein was analyzed by SDS-PAGE and ELISA. RESULTS: The result of SDS-PAGE confirmed that E.coli expressed a 48 kD protein. The ELISA result demonstrated that the cell culture supernatant reacted with Rh+ group O human erythrocytes, but was not recognized by Rh-group O human erythrocytes. CONCLUSION: Expressed Fab fragment has the antigenic specificity for human erythrocytes.

Objective:To explore the association between interleukin(IL)-28B single nucleotide polymorphisms and natural outcome of hepatitis C virus.Methods:The IL-28B rs12979860 locus was genotyped in 266 HCV infected volunteer blood donors(107 spontaneous cleared and 159 chronic infection) and 97 healthy controls using Sanger sequencing assay.The difference in rs12979860 genotypes and allele frequencies between the six groups(107 spontaneous cleared and 159 chronic infection,266 HCV infection and 97 healthy controls,159 chronic infection and 97 healthy controls) were analyzed by statistics.Results:159 HCV chronic infection,107 spontaneous cleared and 97 healthy controls,were shown more CC genotype,accounting for 83.6%,95.3%and 86.6%,respectively, while the CT genotype accounted for 16.4%,4.7%and 13.4%respectively.No TT genotype was found.The CC/CT genotype was not significant difference between HCV infection and healthy controls,chronic infection and healthy controls(χ2=0.204,P=0.652;χ2=0.406,P=0.524),but between chronic infections and spontaneous clearance had statistically significant(χ2=8.474,P=0.004),the frequence of C allele in spontaneous cleared was higher than HCV chronic infection(χ2=7.949,P=0.005).Conclusion: The gene polymorphism of IL-28B rs12979860 is not related to HCV susceptibility,but there are differences in chronic infection and spontaneous cleared,showing the C allelic in favor of HCV spontaneous cleaed.