Objective: To analyze the changes in homologous immunity after RhCE-matched transfusion in positive patients with RhCE blood group antibodies, and to provide precise transfusion strategies for chronic anemia patients. Methods: Patients with chronic anemia in our hospital from January 2020 to March 2024 (continuously receiving blood transfusions for more than 6 months) were enrolled, and 63 cases of unexpected antibody screening positive and identified as RhCE blood group antibodies were selected as the research subjects. The changes in unexpected antibody yield rate after ABO and RhCcDEe isotype blood transfusion were observed. Patients with MNS, Kidd, or Lewis blood group antibodies were screened for corresponding negative donors using monoclonal antibodies for extended typing transfusion based on RhCcEe typing, and the changes in unexpected antibody yield rate after transfusion were observed. Blood group genotyping was performed when serological techniques failed to resolve discrepancies or detect abnormal antigen expression. Results: After RhCcDEe-matched transfusions, RhCE antibodies disappeared in 62 patients, while 1 patient developed anti-Ce. The latter did not develop blood type isotype immunity after receiving RhccEE donor blood. Among the 62 patients, 9 developed unexpected antibodies against other systems: anti-M (4 cases), anti-Mur (2), anti-S (1), anti-Jka (1), and anti-Lea (1). No additional alloimmunization occurred after extended antigen-matched transfusions. A patient with serologically weak e phenotype was genotyped as DCe/DcE, with gene sequencing revealing an 827C>A mutation in exon 6 of the RHCE gene, forming the RHCE 01.31 allele. Conclusion: Precise transfusion strategies incorporating RhCE, MNS, Kidd, and Lewis blood group antigen typing can reduce the probability of blood group homologous immunity. RhCE complex antibodies and RhCE variants pose difficulties for clinical RhCE typing transfusion, which can be addressed through cross-matching and genetic analysis.

Objective: To evaluate the feasibility of exempting Asian-type DEL pregnant women from anti-D monitoring and RhD immunoglobulin prophylaxis injections by comparing and analyzing the clinical incidence of anti-D alloimmunization between Asian-type DEL pregnant women and true RhD-negative pregnant women. Methods: A total of 165 pregnant women who were initially screened as RhD negative by the saline method and received medical treatment in our hospital from December 2022 to August 2024 were collected as the research subjects. Absorption and elution tests, DEL genotyping, and gene sequencing were used to divide the pregnant women into the Asian-type DEL group and the true negative group. After obtaining informed consent, the following clinical management plan was implemented for pregnant women with Asian-type DEL: exemption from routine anti-D antibody detection, exemption from RhD immunoglobulin prophylaxis, and transfusion of RhD-positive red blood cells. Blood samples of newborns were sent for examination of hemolytic disease of the fetus and newborn (HDFN). The routine management plan was implemented for true negative pregnant women. The incidence of alloimmunization and HDFN was comparatively analyzed between the two groups. Results: Among 165 initially screened RhD negative pregnant women, serological testing and genotyping confirmed 42 as Asian-type DEL, 9 as D variant, and 114 as true negative. Among 42 pregnant women with Asian-type DEL, 3 cases tested positive for HDFN due to receiving RhD immunoglobulin prophylaxis injection. The remaining 39 cases were exempted from anti-D testing after being fully informed of the risk, and did not receive RhD immunoglobulin prophylaxis. The HDFN tests were all negative. In the true negative group, anti-D antibodies were detected in 20 cases, of which 6 cases tested positive for HDFN. A pregnant woman with Asian -type DEL did not show RhD homologous immune response after receiving 2 units of RhD positive red blood cells. Statistical analysis revealed a significantly lower risk of anti-D alloimmunization in Asian-type DEL carriers compared to true D-negative pregnant women (P<0.05). Conclusion: Pregnant women with Asian-type DEL can be exempted from routine anti-D antibody testing and do not require routine RhD immunoglobulin prophylaxis injections.

Objective To study the feasibility of detecting fetus RhD type gene by Surface Plasmon Reso-nance(SPR)technology,and to establish a new rapid diagnosis method for fetus RhD type gene.Methods The different types of DNA corresponding RNA probes were fixed on the surface of SPR chip by using amino cou-pling methods,and optimize the chip analysis condition,and then using the RNase H enzyme hydrolysis,signal amplification detection,at last the detection conditions were determined.We use the RhD type gene exon 5,7 of RNA probe to test its corresponding DNA molecules,and analyse the specificity and sensitivity of SPR chip detection signal.Results The SPR technique for detecting the exon 5,7 of RhD blood type gene shows good sensitivity and specificity in all,SPR technology can specifically detect the Exon 5,7 of RhD blood type gene, and the sensitivity of for detecting RhD gene exon 5,7 is 100 pmol/L by SPR.Conclusion The SPR technolo-gy can quickly detect RhD gene accordingly,SPR technology is simple,rapid,reliable and label-free,w hich can provide a new way predicting fetal RhD type for RhD negative prenatal.

Objective To study the platelet antibody screening and crossing match by surface plasmon resonance(SPR) ,and to find a new way for platelet compatibility testing .Methods The corresponding universal platelet antigen was fixed on the SPR chip surface by the amino coupling method .Platelet antibody positive and negative control serum were analysed by SPR micro-ar-ray ,the stability ,sensitivity and specificity of this technique were discussed ,and compared with MAIPA assay .Finally we used the SPR technology to cross match for ten cases of the platelet antibody positive patients before infusion ,and to evaluate the effect of platelet infusion .Results For the SPR technology ,the stability ,sensitivity and specificity of platelet antibody detection were all better ,106 cases of the repeated platelet transfusion samples were tested by SPR assay and MAIPA method ,there was no signifi-cant difference between them(chi-square=0 .333 ,P>0 .05) ,the total consistency was 97 .2% .The 10 cases of platelet antibody positive patients were crossed match before platelet transfusion by SPR technology ,the good results of 8 cases of them were found by the clinical tracking evaluation ,1 h CCI>7 .5 ,24 h CCI>4 .5 .Conclusion SPR technology for screening platelet antibody mat-ches with MAIPA method in basic quality ,but SPR assay is simple ,rapid ,reliable and intuitive ,label free ,which can satisfy the re-quirements for clinical rapid detection of platelet antibody screening and crossing match .

BACKGROUND:As a main antigen of platelet, CD36 antigen is also known as platelet glycoprotein IV (GPIV). The mutation of CD36 gene may result in deficiency of the antigen.
OBJECTIVE:To identify a novel CD36 alele.
METHODS: DNA was isolated from peripheral blood sample, and 12 coding regions of CD36 gene were amplified by PCR. Sequencing-based typing was used to analyze the sequence of the target regions. The derived sequences were aligned with the standard sequence of NG_008192 in GenBank to identify the novel alele.
RESULTS AND CONCLUSION: 1142 T>G mutation was detected in exon 12 of CD36 gene of the proband, and the other regions were consistent with the standard sequence. No data or report about 1142 T>G was found in GenBank or National Center for Biotechnology Information (NCBI), and thus it was reported to GenBank and received by number KM275213. 1142 T>G results in amino acid 381 Leu>Ser of the CD36 protein. There is a big difference in hydrophilia and polarity of the two amino acids. Also the 381 amino acid locates in highly conserved region. Thus it is speculated that 1142 T>G may reduce or vanish the activity of the protein.

OBJECTIVETo determine the frequency of RHD1227A allele in Chinese Hans.

METHODSFor a total of 890403 ethnic Han blood donors, the D antigen was determined with a saline method and indirect antiglobulin test. The RHD1227A allele and number and type of zygosity of RHD gene were determined with PCR sequence specific primer (PCR-SSP). Allelic frequency was calculated through statistics.

RESULTSIn total 2385 donors were found to be Rh-negative, 108 individuals were found to be weakly positive for D antigen (including weak D and partial D phenotypes). The remaining 887 910 individuals were Rh-positive. Among the Rh-negative individuals, 516 were found with RHD1227A. Among these, 467 were RHD1227A/d and 49 were RHD1227A/RHD1227A. Two of 108 D antigen weak-positive individuals were found as RHD1227A/RHD+. In addition, 8 of 1073 random Rh-positive samples were found to be RHD1227A/RHD+. The allele frequency of RHD1227A in the population was calculated as 0.004 036. The figure should be 0.006 682 if calculated based on the detected rate of the allele in Rh-negative individuals, and 0.007 884 if calculated based on the reported average phenotype rate of DEL in Rh-negative individuals.

CONCLUSIONBy taking main influencing factors such as the RHD zygosity, the rate of RHD1227A and DEL phenotype may be determined. The allele frequency of RHD1227A in Chinese Hans is between 0.004 036 and 0.007 884.

Adolescent ; Adult ; Asian Continental Ancestry Group ; ethnology ; genetics ; China ; ethnology ; Exons ; Female ; Gene Frequency ; Genotype ; Humans ; Male ; Middle Aged ; Mutation, Missense ; Rh-Hr Blood-Group System ; genetics ; Young Adult

Objective To study the changes of the peripheral blood regulatory B cells and related cytokines in children patients with chronic idiopathic thrombocytopenic purpura (CITP),and to analyze their correlations with effective Th cells,and to discuss their roles in the pathogenesis of CITP.Methods The peripheral blood mononuclear cells (PBMC)were separated in 30 cases of CITP,then percentages of Th1 ,Th17,Th22 and Breg cells were detected by the flow cytometry(FCM).IFN-γ,IL-17,IL-22 and IL-10 levels in culture supernatant were measured by the ELISA method.Results Compared with the control group,the proportions of the peripheral blood Th1 ,Th17,Th22 subsets in CITP children were increased[(18.4±4.7)% vs.(10.82±4.4)%;(2.42±1.02 )% vs.(1 .23±0.42)%;(0.79±0.24)% vs.(0.26±0.11)%],the proportion of Breg cells was reduced significantly,the differ-ences had statistical significance[(0.83±0.21)% vs.(1 .89±0.12)%,P <0.05].In CITP children,the IFN-γ,IL-17,IL-22 levels in culture supernatant were higher than those in the control group[(278.2±121.2)pg/mL vs.(181.8 ±82.2)pg/mL;(214.8 ± 100.5)pg/mL vs.(161 .4±67.8)pg/mL;(122.16±22.2)pg/mL vs.(90.8±31.1)pg/mL],but the IL-10 level was lower than that in the healthy control group[(27.4±12.6)pg/mL vs.(46.1±16.2)pg/mL),differences between them had statistical signifi-cance(P <0.05).Besides,there was a positive correlation between the proportion of Breg cells and cytokine IL-10 level(r=0.617, P <0.05 ),however,there was a negative correlation between the proportion of Breg cells with Th1 ,Th17 and Th22 cells (P <0.05),between IL-10 level with IFN-γ,IL-17 and IL-22 levels(P <0.05).Conclusion The downregulation of Breg cells proportion may participate in the immune disorder mechanism of effective Th cells in CITP children,which can provide a new idea for the im-munoregulation therapy in CITP children.

The whole length of exon 11 of BRCA1 was sequenced (total 3427 bp) in 59 patients and 10 healthy female blood donors. To allow a rapid determination of the different BRCA1 alleles, a sequence-specific primer PCR method (PCR-SSP) was established and was applied to 57 additional female donors. Finally, the full-length coding region of BRCA1 was analyzed through reversed-transcriptase PCR (RT-PCR) and cDNA sequencing (total 5554 bp) in one donor with wild-type allele and 2 patients with one or two mutated alleles. By genomic DNA sequencing, 5 homozygous polymorphisms were observed in 18 patients: 2201C>T, 2430T>C, 2731C>T, 3232A>G and 3667A>G All of them were previously observed in Caucasians, Malay and Chinese, but for the first time the mutations were found in one allele (GenBank AY304547). Twenty-six patients and 4 donors were heterozygous at these 5 nucleotide positions. The remaining 15 patients and 6 donors showed a sequence identical with the standard BRCA1 gene. Combined the PCR-SSP results and in a summary, 6 of 67 (9.0 %) healthy individuals were homozygous for the mutated allele, whereas 18 of 59 (30.5 %) breast cancer patients were homozygous. A Chi-square test showed a significant correlation between homozygous mutated BRCA1 allele and breast cancer. The cDNA sequencing showed that 2 additional mutations, 4427T>C in exon 13 and 4956A>G in exon 16, were found. A new BRCA1 allele, which is BRCAI-2201T/2430C/2731T/3232G/3667G/4427C/4956G (GenBank AY751490), was found in Chinese. And the homozygote of this mutated allele may implicate a disease-association in Chinese.

The whole length of exon 11 of BRCA1 was sequenced (total 3427 bp) in 59 patients and 10 healthy female blood donors. To allow a rapid determination of the different BRCA1 alleles, a sequence-specific primer PCR method (PCR-SSP) was established and was applied to 57 additional female donors. Finally, the full-length coding region of BRCA1 was analyzed through reversed-transcriptase PCR (RT-PCR) and cDNA sequencing (total 5554 bp) in one donor with wild-type allele and 2 patients with one or two mutated alleles. By genomic DNA sequencing, 5 homozygous polymorphisms were observed in 18 patients: 2201C＞T, 2430T＞C, 2731C＞T, 3232A＞G and 3667A＞G. All of them were previously observed in Caucasians, Malay and Chinese, but for the first time the mutations were found in one allele (GenBank AY304547). Twenty-six patients and 4donors were heterozygous at these 5 nucleotide positions. The remaining 15 patients and 6 donors showed a sequence identical with the standard BRCA1 gene. Combined the PCR-SSP results and in a summary, 6 of 67 (9.0 %) healthy individuals were homozygous for the mutated allele, whereas 18 of 59 (30.5 %) breast cancer patients were homozygous. A Chi-square test showed a significant correlation between homozygous mutated BRCA1 allele and breast cancer. The cDNA sequencing showed that 2 additional mutations, 4427T＞C in exon 13 and 4956A＞G in exon 16, were found. A new BRCA1 allele, which is BRCA1-2201T/2430C/2731T/3232G/3667G/4427C/4956G (GenBank AY751490), was found in Chinese. And the homozygote of this mutated allele may implicate a disease-association in Chinese.