1.Serological and molecular study of a novel B(A) allele with multiple missense mutations.
Bin HAN ; Shaoqiang ZHANG ; Yuli ZHU ; Zhihui FENG ; Shutao PANG
Chinese Journal of Medical Genetics 2021;38(1):20-22
OBJECTIVE:
To explore the molecular basis for an individual suspected as AwB subtype through DNA sequencing.
METHODS:
ABO serology was carried out with the standard tube method. To identify the ABO gene haplotype, the amplicons of exon 7 were cloned and sequenced.
RESULTS:
Serological results showed that the forward typing was AwB and the reverse typing was B. Sequencing analysis revealed that the sample has contained an O01 allele in addition with c.297A>G, c.657C>T, c.796C>A, c.803G>C, c.930G>A variants as compared with the A101 allele.
CONCLUSION
Through sequencing analysis, the sample with an AwB subtype by serological testing was identified as a novel B(A) phenotype, which was unreported previously.
ABO Blood-Group System/genetics*
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Alleles
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Base Sequence
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Exons/genetics*
;
Humans
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Mutation, Missense
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Phenotype
2.Construction and validation of eukaryotic expression vectors for human ABO subgroup genes.
Zhihui FENG ; Bin HU ; Tongxian WANG ; Shuxian JIAO ; Shutao PANG
Chinese Journal of Medical Genetics 2018;35(1):64-68
OBJECTIVE To construct eukaryotic expression vectors for human ABO and subgroup genes A101, B101, CisAB01, Ael05, B(A)04 and Bw03, and validate their expression in vitro. METHODS Total RNA was isolated from individuals with the A101 and B101 subgroups. cDNA of A101 and B101 was synthesized by reverse transcription and amplified with specific primers. Subgroup genes CisAB01, Ael05, B(A)04 and Bw03 were then amplified with PCR for site-directed mutagenesis. Fragments of the ABO genes were directionally linked to pcDNA3.1 positive-eukaryotie expression vectors. After antibiotic screening, the sequences were analyzed. The vectors were transfected into Hela cells, and the expression of target proteins was detected by Western blotting and immunofluorescence assay. RESULTS Sanger sequencing has confirmed that pDNA3.1-A101, pDNA3.1-CisAB01, pDNA3.1-Ael05, pDNA3.1-B101, pDNA3.1- B(A)04, pDNA3.1-Bw03 positive-eukaryotic expression vector were successfully constructed. The results of Western blotting and immunofluorescence revealed clear presence of the expressed proteins. CONCLUSION Eukaryotic expression vectors for ABO subgroup genes were successfully constructed and worked well in Hela cells in vitro, which can facilitate further study of the ABO blood group proteins.
3.Comparative analysis of blood components distribution in 24 domestic prefecture-level blood stations
Cheng PENG ; Guanlin HU ; Li LI ; Zhenxing WANG ; Jinghan ZHANG ; Yugen CHENG ; Liping HUANG ; Qiuhong MUO ; Yang LIU ; Wenzhi WANG ; Haining WANG ; Hao LI ; Youhua SHEN ; Xiaojuan YANG ; Guoqian YANG ; Ling WU ; Feng YAN ; Ning LI ; Jing LIU ; Lin BAO ; Mengshang ZHANG ; Jing CUI ; Zhujun FU ; Helong GUO ; Shutao PANG
Chinese Journal of Blood Transfusion 2022;35(9):942-946
【Objective】 To understand the current situation of blood components distribution in domestic prefecture-level blood stations through analyzing the components distribution data of 24 prefecture-level blood stations in China. 【Methods】 The data of components distribution of 24 blood stations from 2017 to 2020 as well as the data of blood deployment of 24 blood stations from 2019 to 2020 were collected and analyzed. 【Results】 From 2017 to 2020, positive annual growth in red blood cells, plasma and cryoprecipitate was observed in 22, 19 and 15 out of the 24 blood stations, and the annual growth median rate of above three components was 5.24%, 3.80% and 3.25%, respectively. Among the 24 prefecture-level blood stations, 23 carried out the preparation of cryoprecipitate. 【Conclusion】 The distribution of red blood cells, cryoprecipitate and plasma in prefecture-level blood stations is increasing year by year. However, there is a overstock of plasma, and most blood stations need blood employment.