Objective To develop a method to produce leukoreduced platelet concentrates(LR-PCs) from pooled buffy coats in additive solution(a mixture of solutions for medical use).Methods LR-PCs were made from 6 pooled buffy coats(12 whole-blood units) and 220g additive solution,including 90% multiple electrolytes injection solution,8% ACD-A and 2% 50g/L NaHCO3 injection solution.After centrifugation,the PCs were leukoreduced with a filter and stored in a 600ml platelet storage bag.LR-PCs were prepared in a closed system.Results Routinely produced LR-PCs(n=30) contained(2.96?0.31)?1011 platelets with a volume of(270? 32)ml.The WBC and RBC count were(1.3?0.2)?106 and(5.8?1.1)?109 per unit,respectively.The CD62P expression was(22.5? 10.6)%.After 8-day storage,the in vitro quality of LR-PCs,including pH,hypotonic shock response(HSR) and CD62P expression were 7.14?0.04,(54.0?8.2)% and(45.7?13.8)%,respectively.Conclusion In terms of the in vitro quality of LR-PCs,the method of preparing LR-PCs from pooled buffy coats in mixing solutions is feasible.

Objective To investigate the protective effect and its potential molecular mechanism of Nec-1 on cytotoxicity induced by cyclosporine A.Methods MRTEpiC, glomerular endothelial cell MGEC and mesangial cell line MMC were co-administered with Nec-1 and cyclosporin A in mouse renal tubular epithelial cell line, and then MTT assay and soft agar clone formation assay were used to detect Cell growth curve changes, clonal formation ability.Apoptosis was detected by flow cytometry.The expression of cyclin D1, CDK4, CDK2, Cyclin E and apoptosis-related Caspase 3 were detected by Western blot.Results After cyclosporine A action, the cell growth ability was significantly decreased and the clone formation ability was significantly decreased(P<0.05).Cyclin D1, CDK4, CDK2 and Cyclin E were significantly increased(P<0.05), but the ratio of apoptosis and the expression of Caspase 3 did not change.Nec-1 has obvious protective effect on cytotoxicity induced by cyclosporine A, which can increase the cell growth ability and clone formation ability, and reduce the cell cycle-related proteins Cyclin D1, CDK4, CDK2, Cyclin E.Conclusion Nec-1 has cytotoxic effect on the glomeruli and renal tubular cells by up-regulating the cell cycle-related proteins Cyclin D1, CDK4, CDK2 and Cyclin E, while Nec-1 has protective effect.

OBJECTIVETo explore the molecular mechanism underlying the DEL phenotype among RhD negative ethnic Han individuals from Jiangsu, China.

METHODSThe DEL phenotype was determined by an adsorption elution test among 57 RhD negative blood donors. The Rh C, c, E, and e phenotypes were detected by a tube method. PCR with sequence-specific primering (PCR-SSP) assay was used to determine the RHCE genotypes. The RHD gene of the DEL individuals were amplified with polymerase chain reaction and subjected to Sanger sequencing analysis.

RESULTSAmong the 57 RhD negative donors, 10 (17.54%) were determined as having the DEL phenotype. The major RhCE phenotypes for DEL and RhD negative cases were RhCcee (80.0%) and Rhccee (61.7%), respectively. All RHD gene sequences of the 10 individuals have harbored a G>A mutation at position 1227 of exon 9.

CONCLUSIONA proportion of RhD negative individuals determined by routine serological method are actually DEL with RHD gene mutations. RHD *1227A is the most prevalent DEL genotype among ethnic Han Chinese from Jiangsu. Further research on the phenotype and underlying molecular mechanism of DEL is important for blood transfusion.

Alleles ; Asian Continental Ancestry Group ; ethnology ; genetics ; Base Sequence ; Blood Donors ; China ; ethnology ; Exons ; Genotype ; Humans ; Male ; Molecular Sequence Data ; Phenotype ; Polymorphism, Genetic ; Rh-Hr Blood-Group System ; genetics

OBJECTIVETo identify a rare subtype of the ABO blood group system and explore its molecular basis.

METHODSBased on a standard serological assay, ABO subtype and haplotype were analyzed through PCR amplification of the 7 exons and adjacent introns of the ABO gene and TA clone sequencing.

RESULTSForward typing showed a B type, while reverse typing demonstrated an extremely weak anti-B on routine gel analysis, which indicated a forward and reverse typing discrepancy. Absorption-elution testing confirmed that there was no A antigen on the surface of patient's red blood cells. Sequencing of the ABO gene showed a G>A exchange at position 523 in exon 7, which resulted in a Val to Met substitution at codon 175. Clone sequencing of the amplificons of the ABO gene showed an ABO* Bw14/O01 heterozygote genotype.

CONCLUSIONMolecular method is useful for the identification of ambiguous blood groups. A 523G>A substitution of the ABO gene resulting in a Bw14 subtype probably underlies the weak B phenotype noted in the patient.

ABO Blood-Group System ; genetics ; Exons ; Genotype ; Humans ; Male ; Middle Aged ; Phenotype ; Polymerase Chain Reaction

OBJECTIVE: To investigate the serological and molecular characteristics of a pedigree carrying an allele for ABO*BW.11 blood subgroup. METHODS: The ABO blood type of 9 pedigree members were determined by serological methods. Exons 6 and 7 of the ABO gene were amplified by PCR and directly sequenced. The patient and her father were also subjected to clone sequencing analysis. RESULTS: Serological tests demonstrated that the proband and her younger brother had an ABw subtype, whilst her father and two daughters had Bw subtype. Clone sequencing found that the exon 7 of the ABO gene of the proband had a T>C substitution at position 695, which was identified as a BW.11 allele compared with the reference sequence B.01. This BW.11 allele was also identified in the proband's father, brother and two daughters. Due to allelic competition, the A/BW.11 and BW.11/O alleles demonstrated significantly different phenotypes. CONCLUSION The c.695T>C substitution of the ABO gene may lead to allelic competition in the Bw11 subtype. Combined molecular and serological methods is helpful for precise blood grouping.
ABO Blood-Group System/genetics* ; Alleles ; Female ; Genotype ; Humans ; Male ; Pedigree ; Phenotype

ABO Blood-Group System ; genetics ; Exons ; genetics ; Female ; Genotype ; Humans ; Middle Aged