The study was aimed to analyze Del phenotype of RhD (-) unrelated blood donors. RhD (-) was initially screened by routine serological test and confirmed by indirect antiglobulin test (IAT). Del phenotype was detected by hot-ether absorption-elution test. The results indicated that 106 RhD (-) samples were confirmed out of 38526 donors, and 28 cases were Del detected by hot-ether absorption-elution test. The incidence of Del in RhD (-) samples was 26.41%, The serological phenotypes of Del were Ccee (78.57%), CCee (14.29%) and CcEe (7.14%) respectively. In conclusion, the detection of Del by using hot-ether absorption-elution test is very important for reasonable application of RhD (-) blood. There is difference in Del phenotypes of populations in different regions of China and Japan.
Asian Continental Ancestry Group ; Blood Donors ; Humans ; Phenotype ; Rh-Hr Blood-Group System ; genetics ; immunology

OBJECTIVE: To explore the the effects of 2-Me, DTT, papain, pineapple protease and ZZAP on the antigenicity of JMH antigen of human red blood cells (RBC) surface. METHODS: Firstly, human RBC were treated with 2-Me, DTT, pineapple protease, papain and ZZAP reagents, respectively. The antigenicity of JMH antigen on human RBC surface was detected and analyzed by flow cytometry. RESULTS: Flow cytometric analysis found that compared with level before treatment, the antigenicity of JMH antigen on RBC surface was significantly reduced after 2-Me treatment, the positive rate of JMH antigen: 69.5%±4.5% vs 56.5%±3.4% (t=12.44, P＜0.01); fluorescence intensity: 4906±317 vs 3003±165 (t=11.84, P＜0.01). The antigenicity of JMH antigen on RBC surface significantly increased after DTT treatment, showing the positive rate of JMH antigen: 61.7%±3.8% vs 75.5±4.9% (t=16.57, P＜0.01), fluorescence intensity: 4044±294 vs 4854±319 (t=15.46, P＜0.01). However, both bromelain and papain could significantly reduce the antigenicity of JMH antigen on the RBC surface, Bromelain: the positive rate of JMH antigen: 62.2%±3.8% vs 8.8%±1.2% (t=26.44, P＜0.01), fluorescence intensity: 4263±273 vs 1444±212 (t=19.27, P＜0.01); Papain: the positive rate of JMH antigen: 62.8%±3.6% vs 8.8%±1.5% (t=21.38, P＜0.01), fluorescence intensity: 4389±284 vs 1458±230 (t=17.49, P＜0.01). The flow cytometric analysis revealed that ZZAP treatment significantly reduced the antigenicity of JMH antigen on the RBC surface, the positive rate of JMH antigen: 62.2%±4.4% vs 48.2%±4.1% (t=14.87, P＜0.01), fluorescence intensity: 4106±263 vs 2063±175 (t=17.49, P＜0.01). CONCLUSION The treatment with 2-Me can reduce the antigenicity of JMH antigen on human RBC surface. The antigenicity of JMH antigen on human RBC surface increased after DTT treatment. The antigenicity of JMH antigen on human RBC surface significantly reduces after the treatment with pineapple protease or papain. ZZAP treatment can reduce the antigenicity of JMH antigen on the RBC surface.
Blood Group Antigens ; Erythrocytes ; Flow Cytometry ; Humans ; Rh-Hr Blood-Group System

The Rh blood group system is one of the most complex and important systems known in humans. It has two homologous structure genes in tandem on 1p34.3-36.1 that encode Rh protein. The Rh protein is a membrane in red blood cell that has 12 transmembrane spans. Rh antigens have many variants; there are three genetic polymorphisms in the RhD-negative individual. The Rh blood group system is of great significance in clinical transfusion and hemolytic disease of the newborn (HDN). Rh PCR genotyping is used for prenatal diagnosis in fetus, but still it has some defects, and in this connection further knowledge about Rh system will be necessary to solve the problem.
Erythroblastosis, Fetal ; blood ; Humans ; Infant, Newborn ; Rh-Hr Blood-Group System ; blood ; genetics

The efficiency of cold storage red blood cells (CSRBC) or whole blood at -80 degrees C used in 27 Rh (D) negative patients during surgical operation was reported. The Rh (D) negative patients received the transfusion of CSRBC or whole blood stored at -80 degrees C for 180 to 360 days. The changes in the indexes, such as blood TB, DB, K+, Na+, BUN, Cr, urine protein (URPO), UOB, Hb, HCT, serum total protein, relative to hemolytic reaction and blood volume before and after transfusion were observed. The results showed that after transfusion of CSRBC or whole blood 27 cases were negative for urine protein and UOB, and the levels of BUN and Cr were normal (P > 0.05). Blood TB, DB, Hb, and HCT were increased, while pH, blood K+ and blood Na+ was normal with the difference being not significant before and after operation (P > 0.05). Plasma protein was decreased, but there was no significant difference before and after operation (P > 0.05). It was suggested that CSRBC or whole blood at -80 degrees C could be safely infused to the Rh(D) negative patients without side effects during the surgical operation.
Blood Preservation/*methods ; Blood Transfusion/*methods ; *Cryopreservation ; Erythrocytes ; Intraoperative Care ; *Rh-Hr Blood-Group System

OBJECTIVETo study the molecular mechanism and polymorphism of D gene of RhD negative and D variants among voluntary blood donors from Qingdao region.

METHODSFor 220 D-negative phenotype cases and 5 D variant cases confirmed by serological test, exons 1 to 10 of the RHD gene were detected by a PCR-SSP method. The samples which contain all or part of the exons were sequenced.

RESULTSAmong the 220 cases, 166 (75.45%) had complete absence of the RHD gene, while 54 (24.55%) had retained some or all of the 10 exons. Eight genotypes were identified, which included RHD 1227G>A in 28 cases (12.73%), RHD-CE- (2-9) -D in 19 cases (8.64%), RHD-CE- (3-7)-D in 1 case (0.45%), RHD 3G>A in 1 case (0.45%), RHD 711delC in 2 cases (0.91%), RHD 845G>A in 1 case (0.45%), RHD 1013T>C in 1 case (0.45%), and RHD 1227A/G in 1 case (0.45%). No mutation was found in all of the 10 exons. Two alleles were identified in the 5 cases of D variants, which included RHD 845G>A (4 cases) and RHD 697G>A (1 case).

CONCLUSIONAbsence of the whole RHD gene is common among RhD negative blood donors from Qingdao region, and there are rich genetic polymorphisms for this locus.

This study was purposed to establish the method of quantifying RhD antigen on red blood cells (RBC) by flow cytometry (FCM) and to explore the expression of D antigen on RBC of different RhD serotype. RhD(+) RBCs and RhD(-) RBCs were mixed in 1:1 ratio. Cells were stained by the indirect method (IgG anti-D as the first antibody, FITC-anti-IgG F(ab')2 as the second antibody), and the ratio of RhD(+) on RBCs was quantified by FCM. The optimal dosage of IgG anti-D was defined. Expression of RhD antigen on RBC of RhD(+), weak D, RhDel and RhD(-) type were detected by FCM. The results showed that optimal dilution of IgG anti-D monoclonal antibody was 1:4, 1x10(6) cells/50 microl. The percentage of D(+) RBC of RhD(+), weak D, RhDel and RhD(-) type were 96.8+/-2.97%, 79.5+/-9.88%, 47.8+/-11.43%, 3.7+/-2.96%, respectively. The mean fluorescence intensity (MFI) of RhD antigen expression of RhD(+), weak D, RhDel and RhD(-) type were 33.3+/-6.21 Dal, 18.6+/-5.39 Dal, 7.10+/-1.17 Dal, 0.79+/-0.55 Dal, respectively. In conclusion, there are significant differences of RhD antigen expressions among RBC of different RhD serotypes. The level of antigen on RhD(+) RBC is the highest and then weak D the next, while the level of antigen on RhDel RBC is the lowest level.
Blood Donors ; Erythrocytes ; immunology ; metabolism ; Flow Cytometry ; methods ; Humans ; Rh-Hr Blood-Group System ; immunology ; metabolism

OBJECTIVE: To investigate the molecular mechanism of one patient with abnormal serological phenotype in RhD and discuss the transfusion strategy. METHODS: The RhD variant sample was screened from a patient with IgM type anti-D antibody and further determined by three different sources of anti-D antibodies. Ten exons and the adjacent introns of the RHD gene were amplified, purified and sequenced. RhCE phenotypes and RHCE genotypes were detected. RESULTS: The patient with Rh variant showed abnormal results of serological tests. The RHD gene sequence analysis showed that the RHD*01W.01 with a variation (c.809T>G, p.Val270Gly) in exon 6 of the RHD gene was found in the patient. The RhCE phenotype was CcEe. The genotyping results of RHCE were consistent with the serological typing results. CONCLUSION The Rh variant of the patient is RHD*01W.01, these findings indicate that RhD variants should be analyzed by molecular assays for the sake of safe transfusion.
Alleles ; Blood Transfusion ; Exons ; Genotype ; Humans ; Phenotype ; Rh-Hr Blood-Group System/genetics*

OBJECTIVE: To analyze the RHD genotype of a blood donor with Del phenotype in Yunnan. METHODS: Rh serological phenotype was identified. RHD gene was detected by PCR-SSP typing, and its 10 exons were sequenced. Exon 9 was amplified for sequencing and analysis. RHD zygosity was detected. RESULTS: The Rh phenotype of this specimen was CcD^elee. Genomic DNA exhibited a 1 003 bp deletion spanning from intron 8, across exon 9 into intron 9. The deletion breakpoints occurred between two 7-bp short tandem repeat sequences. There was no variation in the sequences of the remaining exons. The Rh hybridization box test showed that there was one RHD negative allele. CONCLUSION This specimen is Del type caused by deletion of RHD exon 9.
Humans ; Blood Donors ; Rh-Hr Blood-Group System/genetics* ; China ; Phenotype ; Exons ; Genotype ; Alleles

OBJECTIVETo comprehend the Rh blood type distribution and the gene frequency in four main kinds of the minority nationalities in Guizhou, China, so as to supply a scientific foundation for guiding the prevention and cure against the Rh hemolysis illness.

METHODSThe irrelative individual blood of the four kinds of the minority nationalities (Miao, Buyi, Dong and Shui) in Guizhou was randomly collected in complete group style and the Rh blood type was identified and analyzed.

RESULTSTotally 15,992 minority nationality people were inspected and checked, and 4851 persons of Han nationality people were chosen as controls. Fifty-one persons were proven as Rh (D)negative individuals, in which, d gene frequency of Miao nationality population was 0.0474 and the Rh negative rate was 0.22%, d gene frequency of Buyi nationality population was 0.0602 and the Rh negative rate was 0.36%, d gene frequency of Dong nationality population was 0.0378 and the Rh negative rate was 0.14%, d gene frequency of Shui nationality population was 0.0307 and the Rh negative rate was 0.09%. d gene frequency of Han nationality population was 0.0574 and the Rh negative rate was 0.33%. In the four minority nationality populations, there was a common characteristic of the highest percentage of expressional type, CCDee type (52.47%-59.66%). At the same time, in the gene frequency of the four minority nationality populations, the CDe frequency was the highest: Miao nationality 0.7244, Buyi nationality 0.7389, Dong nationality 0.7410 and Shui nationality 0.7743.

CONCLUSIONThe Rh blood type distribution characteristic of the four minority nationality population, Miao, Dong, Buyi and Shui in Guizhou, is similar to that of the minority population in Southern China. The Rh negative rate of Miao, Dong and Shui nationality populations is lower than that of the Han nationality population(0.33%), only the Rh negative rate of Buiyi nationality population is closer or even higher than that of the Han nationality population. So that,the hemolysis illness frequency in the Rh newborn baby of Guizhou minority nationality population is not high.

The study was to investigate the characteristics of Rh blood group of Uygur nationality in Xinjiang. 1 230 blood samples of Uygur nationality were studied by Rh serological typing, modified antiglobulin test, chloroform/trichloroethylene absorption elution test, upstream, downstream and hybrid Rhesus boxes, 10 exons of D gene, RHD(psi) pseudogene. The results showed that the frequency of RHD negative was 5.8%, and no Del type was found. The further investigation of 72 samples of RhD (-) found that ccee (57.02%) and Ccee (29.08%) phenotype as well as RHD(-)/RHD(-) genotype (94.44%) and complete deletion type of D gene exon (91.12%) were all in high frequency, no RHD(psi) pseudugene was detected. In conclusion, the Rh blood group of Uygurs nationality in Xinjiang possesses both oriental and caucasian Rh characteristics, which enriches the diversity of blood types in chinesenation.
Asian Continental Ancestry Group ; genetics ; China ; ethnology ; Gene Deletion ; Humans ; Rh-Hr Blood-Group System ; genetics