Adverse reactions to blood transfusion: A retrospective analysis of 814 cases in Qingdao area from 2020 to 2021 / 中国输血杂志

【Objective】 To investigate the incidence and composition of adverse reactions to blood transfusion(ARBT) in Qingdao hospitals in recent years. 【Methods】 The "Statistical Table of Adverse Reactions to Blood Transfusion in Medical Institutions" issued by Qingdao Quality Control Center of Blood Transfusion throughout 2020 to 2021, involving 96 hospitals in the region, were collected and analyzed, including the number, proportion and types of ARBT, as well as the types of blood components transfused. 【Results】 From 2020 to 2021, 296 676 cases of blood transfusion in 96 hospitals occurred, and the incidence of ARBT was 0.27% (814/296 676), of which the incidence of ARBT involving plasma transfusion was 0.17% [accounting for 39.07% (318/814) of all transfusion reactions], involving platelet transfusion was 0.68% [31.08% (259/814)], involving erythrocyte transfusion was 0.11% [27.64% (225/814)] and cryoprecipitation transfusion 0.03% [1.47% (12/814)]. The types of ARBT were anaphylaxis 77.64% (632/814), fever 19.78% (161/814), transfusion-related dyspnea 1.47% (12/814), transfusion-related circulatory overload 0.37% (3/814), purpura 0.25% (2/814) and transfusion-related hypotension 0.25% (2/814), delayed hemolysis 0.12%(1/814) and acute hemolysis 0.12%(1/814), respectively. 【Conclusion】 In recent years, the incidence of ARBT in local medical institutions is lower than that of domestic general level, and the main reactions are anaphylaxis and fever following the transfusion of plasma or (and) platelets.The monitoring and control of ARBT should be strengthened in each hospital with accurate and timely report, and active preventive measures should be taken to control or reduce the incidence of ARBT effectively.

Nucleic acid technology(NAT) testing for blood screening: A comparative study on individual donation and minipool NAT test / 中国输血杂志

【Objective】 To compare the detection performance of Cobas s201, a minipool(MP) nucleic acid test(NAT) system, and Panther, a individual donation(ID) NAT system, in blood donor screening. 【Methods】 NAT was conducted on 126 359 blood samples, and initially reactive (IR) samples were either discriminated or resolved by ID testing.The non-discriminated reactive (NDR) samples implicated in Panther sysytem were subjected to ID-NAT by Cobas s201. Some non-repeatable reactivet(NRR) and repeatable reactive (RR) samples implicated in Cobas s201 system were subjected to ID-NAT by Panther. 【Results】 61 MP-IR cases were implicated in a total of 85 128 samples that detected by Cobas 201, and 29(0.34‰) were RR after resolved by ID testing. 74(1.79‰)IR samples were implicated in 41 231 samples that detected by Panther, and 22 (29.73%) were DR-HBV after discriminatory test. Among the NDR 28 samples detected by Panther multiplex system, 7 were positive by Cobas s201 single sample (PP1) whereas non-reactive in simulated MPs of six by Cobas 201.In 28 RR samples resolved by Cobas 201, 24 positive and 4 negative samples were retested by Panther. Among the 11 samples presenting inconsistent retest results by Panther and Cobas 201, 10 were anti-HBc positive, carrying low viral load HBV. 【Conclusion】 The NAT-yield by Panther was significantly higher than that by Cobas s201. Some samples with negative discriminatory results were OBI, and it is necessary to further track and verify the unidentified samples. Cobas s201 is more suitable for a wide array of MP-NAT testing while Panther sample loading, which is flexible and easy to operate, is more suitable for ID-NAT with medium sample size.

Construction and validation of eukaryotic expression vectors for human ABO subgroup genes / 中华医学遗传学杂志

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.

Identification of novel HLA alleles：HLA-A*24:233 and HLA-A*26:89 / 中国组织工程研究

BACKGROUND:The discovery of novel HLA aleles is accelerated by development of molecular biology and applications of numerous new technologies. These new findings not only rich HLA family, but also find a breakthrough point for the study of genetic superiority or disappearance of national gene. OBJECTIVE:To identify two novel HLA-A aleles and to analysis their nucleotide sequences. METHODS:The two samples from two volunteers of Chinese Marrow Donor Program were detected using PCR-SBT and GSSP methods. The HLA-A locus in the two samples were both abnormal genes, and the nucleotide sequence differences were analyzed. RESULTS AND CONCLUSION: The sequences of the samples were different from al aleles in the HLA databases. Sample 1 was found to be different from the closet matching alele A*24:02:01 in one nucleotide substitutions, 360 G > C in exon 3, resulting in amino acid changed from glutamine to histidine at codon 96; and sample 2 differed from A*26:01:01 in one nucleotide substitution, 97 T > C in exon 2, resulting in amino acid changed from tyrosine to histidine at codon 9. The novel aleles were identified and assigned the name HLA-A*24:233 and HLA-A*26:89 officialy by the WHO Nomenclature Committee.

Intracellular Ca2+is involved in survival, proliferation and differentiation of bone marrow-derived mesenchymal stem cells into hepatocytes / 中国组织工程研究

BACKGROUND:The mechanism of differentiation and proliferation of bone marrow-derived mesenchymal stem cells remains unclear. In addition, issues such as how signal pathways such as Ca2+and bone marrow-derived mesenchymal stem cellproliferation and differentiation signals form complex signal network remain poorly understood. OBJECTIVE:To investigate the effect of Ca2+in the induced differentiation of bone marrow-derived mesenchymal stem cells into hepatocytes. METHODS:Bone marrow-derived mesenchymal stem cells were isolated from rat bone marrow using whone bone marrow adherence method, purified, amplified, and induced with hepatocyte growth factor. [Ca2+]i in the directional differentiated bone marrow-derived mesenchymal stem cells and control bone marrow-derived mesenchymal stem cells were detected with flow cytometry. Bone marrow-derived mesenchymal stem cells induced with hepatocyte growth factor were mixed with nimodipine of different concentration, and cells were divided into three groups:hepatocyte growth factor+nimodipine 10 mg/L, 50 or 100 mg/L groups. cellgrowth was observed with inverted phase contrast microscope and alpha 1-antitrypsin expression of the cells was confirmed by immunocytochemistry. The calcineurin M and the activation of extracellular signal regulated kinase pathway was detected by reverse transcription-PCR and western blotting, respectively. RESULTS AND CONCLUSION:[Ca2+]i in the directional differentiated bone marrow-derived mesenchymal stem cells was higher than in the control group (P0.05). These findings indicate that Ca2+could participate in the differentiation of bone marrow-derived mesenchymal stem cells into hepatocytes incuded with cytokines, and also maintain the survival and proliferation of bone marrow-derived mesenchymal stem cells.

Detect and characterize platelet-reactive antibodies in patients who were refractory to platelet transfusions / 中国输血杂志

Objective To detect and determine the specificity of platelet-reactive antibodies in patients who were refractory to platelet transfusions.Methods Serum samples from 48 patients who were refractory to platelet transfusions were screened with MACE for platelet-reactive antibodies.Specificity of platelet alloantibodies was determined with PAK12 and MAIPA.Results Platelet-reactive antibodies were detected in the serum of 50% PTR patients(24/48).The incidence of HLA antibodies was 39.6%(19/48),accounting for 79.2% of serum with platelete alloantibodies.The HPA alloantibodies were found in 29.2%(14/48)serum,of which,64.3%(9/14)occurred together with anti-HLA.The following platelet-specific antibodies were identified:anti-HPA-3a(n=2),anti-HPA-5b(n=1),anti-HPA-5a(n=1),anti-HPA-2b(n=1),anti-HPA-4b(n=1).Of the 14 serum with HPA antibodies,78.6%(11/14)contained panreactive anibodies against platelet glycoprotein(GP)Ⅱb/Ⅲa,GPⅠa/Ⅱa,and/or GPⅠb/Ⅸ.Platelet-reactive antibodies were detected more in female(16/29)than in male(8/19)with a frequency of 55.2%,42.1%,respectively,but there was no statistical significant difference.Conclusion The platelet-specific antibody in PTR patients are not as rare as previous thought although alloantibodies are predominantly anti-HLA.Antibody specificities in Chinese PTR patients are different from those observed in Caucasians,in whom anti-HPA-5b and-1b are the most prevalent specificity.The most prevalent platelet-specific antibodies are anti-HPA-3 and anti-HPA-5 while anti-HPA-4b and anti-HPA-2b are also detected.

Genetic polymorphism of HPA-1 to-5,HPA-15 alloantigen system in Qingdao Han population / 中国输血杂志

Objective To study the polymorphism of human platelet antigen HPA-1 to HPA-5,and HPA-15 system in Qingdao Han population.Methods A total of 918 samples from regular voluntary platelet donors in Qingdao were genotyped for HPA-1 to-5 and HPA-15 by PCR-SSP.Results The gene frequencies of HPA-1a,-1b;HPA-2a,-2b;HPA-3a,-3b;HPA-4a,-4b;HPA-5a,-5b;HPA-15a,-15b were 0.9940,0.0060;0.9319,0.0681;0.5822,0.4178;0.9897,0.0104;0.9804,0.0196;0.4913,0.5087,respectively.Both a and b alleles were found in each of the 6 HPA systems,and a/a homozygosity was more common in HPA-1,-2,-4 and-5 systems.The HPA genotype frequencies followed Hardy-Weinberg principle.HPA-1 frequency of Qingdao people was significantly different from that of North China(P