Objective To investigate the impact of human leukocyte antigen(HLA)functional epitope mismatch(EM)on the efficacy of platelet transfusion in patients with hematological diseases.Methods HLA genotyping was performed on platelet donors and patients with hematological diseases who applied for platelet serological cross-matching and HLA antigen matching from June 2021 to June 2023 by PCR-SBT method.HLA platelet matching was based on the principle of CREG to se-lect donors for patients.HLA Matchmaker 4.0 software was used to analyze donor-recipient HLA EM information.The expres-sion level and gene distribution of related HLA functional epitope(Eplet)were searched from the international HLA Epitope registry website(www.Epregistry.com.br).Retrospective analysis was conducted on clinical platelet transfusion data to evalu-ate the impact of HLA EM on platelet transfusion effectiveness.Results Platelet transfusion efficacy showed no correlation with gender and age,but it was associated with platelet matching strategy.When the total number of HLA EMs was less than 20,a lower total number of donor-recipient HLA EMs resulted in higher platelet transfusion efficiency(χ2=19.311,P=0.001)and higher average value of 24 h corrected count increment(CCI)(F=7.737,P<0.001).The total number of donor-recipient HLA EMs had negative correlation with actual 24 h CCI(Rho=-0.322,P<0.001).Further statistical analysis re-vealed that 17 Eplets were related to the effectiveness of platelet transfusion.The locus distribution of 17 Eplets might be u-nique to HLA-A(17.6%)or-B(64.7%)or shared between HLA-A and-B(17.6%),and its expression may be high(58.8%)or intermediate(41.2%).Conclusion The total number of donor-recipient HLA EMs is a crucial factor influencing platelet transfusion effectiveness,and several HLA Eplets associated with this effectiveness have been identified.

OBJECTIVE: To explore the molecular basis of two individuals with weak D variant of the Rh blood type. METHODS: Routine serological testing was carried out to detect the D, C, c, E and e antigens of the Rh blood group. The D antigen was further detected with an indirect antiglobulin test. The presence of Rhesus box was detected by PCR to determine the homozygosity of the RHD gene. RESULTS: Both samples were determined as weak D phenotype by the indirect antiglobulin test. DNA sequencing revealed that case 1 harbored a heterozygous 208C>T variant in exon 2 and a heterozygous 1227G>A variant in exon 9; while case 2 harbored homozygous 779A>G variants of exon 5 of the RHD gene. Case 1 was determined as RHD+/RHD+, while case 2 was determined as RHD+/RHD-. The two samples were respectively named as weak D type 122 and weak D type 149 based on the rules of Rhesus Base Nomenclature. CONCLUSION D negative blood donors should subject to indirect antiglobulin testing and molecular analysis for safer transfusion.
Alleles ; Blood Donors ; Blood Grouping and Crossmatching ; Genotype ; Humans ; Molecular Biology ; Phenotype ; Rh-Hr Blood-Group System/genetics*

OBJECTIVE: To explore the molecular basis for an individual with para-Bombay phenotype of the H blood group. METHODS: Intron 5 to 3'-UTR of the ABO gene and exon 4 of the FUT1 gene were amplified with PCR and subjected to direct sequencing. Mutations of the FUT1 gene were identified by TOPO cloning sequencing. RESULTS: Direct sequencing showed that her ABO genotype was B101/O01. TOPO cloning sequencing found that this individual had three mutations of the FUT1 gene, including an heterozygous AG deletion (CAGAGAG→CAGAG) at position 547 to 552, and two C→T mutations at positions 35 (C35T) and 293 (C293T) on the other homologous chromosome. The two alleles comprised a new recombination of mutations c.35T>C and c.293C>T, and the sequence has been submitted to NCBI (No. MG597611). CONCLUSION A novel combination of FUT1 alleles with c.35 C>T and c.293C>T has been identified in an individual with para-Bombay phenotype.
ABO Blood-Group System ; Alleles ; Female ; Fucosyltransferases ; genetics ; Genotype ; Humans ; Phenotype

OBJECTIVE: To report on a novel weak D type identified in a Chinese individual. METHODS: Peripheral blood sample was collected for a voluntary blood donor with weakened expression of D antigen. Routine serological testing was carried out to determine the D, C, c, E and e antigens of the Rh blood group. A D-screening kit was used to analyze the RhD epitopes. The 10 exons and flanking intronic regions of the RHD gene were sequenced. The zygosity of RHD was determined with a sequence-specific primer PCR method. RESULTS: A novel RHD allele, RHD (1022T>A), was found in the subject with a weak D phenotype. Serological testing of the RhD epitopes has coined with the weak D phenotype. CONCLUSION A novel weak D allele has been identified in Chinese population.
Alleles ; Asian Continental Ancestry Group ; China ; Exons ; Genotype ; Humans ; Introns ; Rh-Hr Blood-Group System ; genetics

OBJECTIVE: To explore the molecular basis for an individual with Ax28 phenotype of the ABO subtype. METHODS: The ABO group antigens on red blood cells of the proband were identified by monoclonal antibodies. The ABO antibody in serum was detected by standard A, B, O cells. Exons 1 to 7 of the ABO gene were respectively amplified by PCR and directly sequenced. Amplicons for exons 5 to 7 were also sequenced after cloning. RESULTS: Weakened A antigen was detected on red blood cells from the proband. Both anti-A and anti-B antibodies were detected in the serum. Heterozygous 261G/del was detected in exon 6, while heterozygous 467C/T and 830T/C were detected in exon 7 by direct DNA sequencing. After cloning and sequencing, two alleles (O01 and Ax28) were obtained. Compared with A102, the sequence of Ax28 contained one nucleotide changes (T to C) at position 830, which resulted in amino acid change (Val to Ala) at position 277. CONCLUSION The novel mutation c.830T>C of the galactosaminyltransferase gene may give rise to the Ax28 phenotype.
ABO Blood-Group System ; genetics ; Alleles ; Amino Acid Substitution ; Exons ; Galactosyltransferases ; genetics ; Genotype ; Humans ; Phenotype ; Polymorphism, Single Nucleotide ; Sequence Deletion

Objective To analyze the effects of different therapies on patient survival,to explore the related prognostic factors in elderly patients with advanced gastric cancer,and to provide recommendations for the treatment of such patients.Methods Retrospective analysis was conducted on 146 elderly patients with advanced gastric cancer hospitalized from January 2009 to October 2013 in Yixing People's Hospital of Jiangsu Province.Detailed clinical data were recorded,and patients were followed up during the total survival time.Univariate analysis with the Log rank test and multivariate analysis with the COX proportional hazard model were utilized to examine the related prognostic factors in elderly patients with advanced gastric cancer.Results The 1-,2,3-year survival rates of t46 elderly patients with advanced gastric cancer were 33.6 ％,11.0 ％,and 2.1 ％,respectively,and the median survival time was 10.3 months.The Log-rank test showed that Karnofsky (KPS) score,differentiation degree,number of metastatic sites,malignant serous effusion,chemotherapy,and traditional Chinese medicine (TCM) were associated with the prognosis of elderly patients with advanced GC (all P＜0.05).Multivariate analysis by the COX proportional hazard model showed that KPS score (HR=1.575,95％ CI:1.094 2.267,P=0.015),differentiation degree (HR=0.499,95％CI:0.340-0.732,P＜0.001),malignant serous effusion (HR=0.516,95％ CI:0.356-0.748,P＜ 0.001),chemotherapy (HR=1.669,95％ CI:1.185 2.351,P=0.003),and TCM (HR=1.793,95％ CI:1.237-2.600,P=0.002) were independent factors related to the prognosis of elderly patients with advanced GC.Conclusions The prognosis of elderly patients with advanced gastric cancer is poor,especially for patients with a low KPS score,a poor differentiation degree,or malignant serous effusion.Chemotherapy and TCM can improve the prognosis.

OBJECTIVETo identify a novel Bx13 allele.

METHODSSerological characteristics was determined with standard serological methods. All of the seven exons and flanking regions of the ABO gene were analyzed with PCR and direct sequencing. The amplicon of exon 7 was also cloned and sequenced.

RESULTSThe individual was determined as with a rare Bx phenotype by serological tests. Direct DNA sequencing showed that the individual was heterozygous for the B/O01 allele, while there was a novel 893C>T mutation in the B101 allele, which has led to an amino acid substitution Ala298Val in the α,3-D-galactosyl-transferase. The mutation was not found among 100 randomly selected blood donors.

CONCLUSIONA novel Bx13 allele has been identified. Substitution of amino acid in the conserved region of the enzyme may reduce the activity of α,3-D-galactosyl-transferase.

Objective To analyze the effects of valproic acid(VPA),a histone deacetylase(HDAC)inhibitor,on macrophage polarization in?duced by paraquat(PQ)or lipopolysaccharide(LPS). Methods Mouse RAW264.7 cells were cultured at 37℃with 5%CO2,passaged,and then given one of the following treatments:(1)PQ;(2)PQ+VPA(classⅠandⅡa HDAC inhibitor);(3)PQ+apicidin(classⅠHDAC inhibitor);(4)PQ+MC1568(classⅡa HDAC inhibitor);(5)LPS;(6)LPS+VPA;(7)LPS+apicidin;(8)LPS+MC1568. The cells and culture supernatants were harvested after 8 h of treatment. RT?PCR,ELISA,and flow cytometry were conducted to assess the expression levels of macrophage phenotyp?ic markers. Results Both PQ and LPS skewed the macrophage functional polarity toward proinflammatory phenotype. VPA,apicidin,and MC1568 all inhibited PQ?and LPS?induced macrophages polarizing toward pro?inflammatory phenotype ,but the inhibitory effects were different in some ways. Conclusion VPA inhibits the proinflammatory function of macrophages induced by PQ and LPS ,but the effect of VPA on PQ?and LPS?induced macrophages has its own characteristics.

Objective To investigate whether the effect of transient high glucose on inflammatory factors expression could be continuous in rat glomerular mesangial cell,and its relation with histone methylation modification.Methods Rat glomerular mesangial cells (HBZY-l) were divided into three groups:the high glucose group (25.0 mmol/L glucose),the hypertonic group (MA,5.5 mmol/L glucose+ 19.5 mmol/L mannitol) and the normal-glucose control group (5.5 mmol/L glucose),which were cultured for 24 h respectively.All 3 groups were then changed with normal-glucose medium to culture for 24 h,48 h and 72 h.Their protein,mRNA and supernatant were harvested.The protein expressions of mono-methylation of H3 lysine 4 (H3K4mel) was measured by Western blotting,and the mRNA expressions of NF-κB subunit p65 and set7/9 were determined by real timequantitative PCR.The expression of monocyte chemoattractant protein 1 (MCP-1) and vascular cell adhesion molecule 1 (VCAM-1) were detected by enzyme-linked immunosorbent assay.Results (1)Compared with those in normal control group,the expressions of H3K4mel protein and set7/9 mRNA were first up-regulated in high glucose group,then gradually down-regulated in the following 48 h normal-glucose medium (as compared with those at 0 h,all P ＜ 0.05).At 72 h there was no statistic difference between high glucose group and normal control group (all P ＞ 0.05).(2) Compared with those in normal control group,the up-regulated p65 mRNA,VCAM-1 and MCP-1 sustained at least for 72 h in high glucose group.Conclusions Transient high glucose can induce persistent inflammatory factors expression in rat glomerular mesangial cells,which may via histone modification.

OBJECTIVETo explore the molecular basis of 4 cases with weak D variant of Rh blood type.

METHODSRoutine serological testing was applied to determine the D, C, c, E and e antigens of the Rh blood group. The D antigen was further detected with an indirect antiglobulin test. RHD zygosity was detected by sequence-specific primer PCR method. All exons and flanking intron regions of the RHD gene were sequenced.

RESULTSThe samples were determined as weak D phenotype by serological testing. DNA sequencing showed that the 4 cases were heterozygous for 17C>T mutation in exon 1, 29G>C mutation in exon 1, 1212C>A mutation in exon 9, and IVS4+5G>A mutation in intron 4 of the RHD gene, respectively. According to the rule of Rhesus Base Nomenclature, the 4 samples were respectively named as weak D type 31, weak D type 71, weak D type 72, and weak D type 82.

CONCLUSIONSerological and molecular testing for the weak D can facilitate in-depth understanding of its immunology and genetics, and provide guidance for clinical blood transfusion and prevention of hemolytic disease in newborns.