Objective To investigate the allele and specificity distribution situation of HLA-B15 group and HLA-B40 group antigens among Han population in Guangxi area and to explore their possible influence on transplantation donors selection in clinic.Methods The blood samples of 1 644 Han donors in Guangxi region were performed the HLA-B genotyping by PCR-SBT,the frequencies of each allele were calculated by the direct computing method.The antigen specificity of various alleles were analyzed,then the gene frequencies of HLA-B15 and HLA-B40 groups were compared with those from other populations.Results The gene frequency at HLA-B locus in 1 644 Han persons was inconsistent with the Hardy-Weinberg equilibrium(P<0.05).Fourteen alleles in HLA-B15 group were detected out,which belonged to 5 kinds of antigen specificity.In the HLA-B40 group,6 alleles were detected out,which belonged to two kinds of antigen specificity.Conclusion The antigen polymorphism of HLA-B15 and HLA-B40 groups among Han population in Guangxi area is close to that in southern Chinese Han populations,but which still keeps its characteristics of Guangxi area.

Objective To investigate the ambiguity results distribution of HLA-A,B and DRB1 gene sequence-base typing in Guangxi population and to propose the way to resolve.Methods HLA-A,B and DRB1 genes of 1 000 donors in the Guangxi branch bank of China'bone marrow bank were genotyped by PCR-SBT,and then the ambiguity results distribution of the three loci was analyzed.The typing ambiguities resultswere resolved by high-resolution polymerase chain reaction-sequence-specific primers(PCR-SSP) and group specific sequencing primer(GSSP) methods,respectively.Results Among 1 000 samples,at least 1 locus in HLA-A,B and DRB1 genes in 96.7％ samples appeared the ambiguity results,in which the proportions of HLA-A,B and DRB1 loci appearing ambiguity results were 65.7 ％,58.8 ％ and 77.2 ％ respectively.For the samples of detected ambiguity results,single using the GSSP method could resolve the ambiguity typing results of 87.37％ HLA-A,93.54％ HLA-B and 60.49％ HLA-DRB1,using high-resolution PCR-SSP could resolve the ambiguity typing results of 12.63 ％ HLA-A,4.76 ％ HLA-B and 15.29 ％ HLA-DRB1,and the rest 1.70 ％ HLA-B and 24.22 ％ HLA-DRB1 ambiguity results were resolved by both GSSP and high-resolution PCR-SSPs method.Conclusion GSSP and high-resolution PCR-SSPs methods have high abilities to solve HLA ambiguity results both locate inside and outside the sequencing region,respectively.GSSP and high-resolution PCR-SSPs methods are supplement for each other,which can effectively resolve the problem of ambiguity results in high resolution HLA typing.

OBJECTIVETo report on a novel human leukocyte antigen (HLA) allele.

METHODSPolymerase chain reaction-sequence based typing was used for routine HLA typing. For one sample, the result of B locus typing showed mismatch of one base with B*46:01:01, B*15:25:01 at locus 384. The group specific sequencing primers, which target at B*46 and B*15, were used to confirm the difference between the novel allele and the highest homologous allele.

RESULTSThe sequencing results showed that the highest homologous allele to the novel allele was B*46:01:01. The two sequences only differed for position 384 within the exon 3 (384G>T), which resulted in a codon change (GGG>GGT), though the amino acid sequence of the novel allele at position 104 was still Glycine (G). Investigation of the family showed that the novel allele was inherited from the father.

CONCLUSIONThe novel HLA-B allele, discovered in ethnic Zhuangs from Guangxi, has been designated as HLA-B *46:01:18 by the World Health Organization (WHO) HLA Nomenclature Committee.

Adult ; Alleles ; Asian Continental Ancestry Group ; genetics ; Base Sequence ; China ; Exons ; Female ; HLA-B Antigens ; genetics ; Humans ; Male ; Molecular Sequence Data ; Young Adult

In this study, the mature peptide sequence of a pectin lyase gene A was amplified from Aspergillus niger strain EIM-6 by using RT-PCR reverse transcription technique. The cloned gene was then inserted into a Pichia pastoris expression vector pPIC9k to produce the recombinant expression plasmid pPIC9K-pelA. By using electric shocks, we successfully transformed the recombinant pPIC9K-pelA into Pichia pastoris GS115. The activity of the engineered strain reached to 2.3 U/mL after induction with the final concentration of 1.5% methanol. SDS-PAGE analysis revealed that the pPIC9K-pelA transformant had an additional protein band of approximately 38 kD, which was not present in the control. There were no significant differences between the recombinant and native pectin lyase with regard to their hydrolysis activities.
Aspergillus niger ; enzymology ; genetics ; Electroporation ; Pichia ; genetics ; metabolism ; Polysaccharide-Lyases ; biosynthesis ; genetics ; Recombinant Proteins ; biosynthesis ; genetics ; Reverse Transcriptase Polymerase Chain Reaction

【Objective】 To investigate the correlation and consistency between the parameters of thromboelastography(TEG) and routine coagulation tests, and to evaluate the application value of the two methods in heparin anticoagulation monitoring and coagulation function monitoring in patients receiving extracorporeal membrane oxygenation(ECMO) therapy. 【Methods】 A total of 138 patients who recieved ECMO in the Department of Critical Care Medicine of the People′s Hospital of Guangxi Zhuang Autonomous Region from October 2021 to December 2022 were selected. A total of 317 pairs of ordinary TEG and heparinase-modified TEG(hmTEG) parameters measured simultaneously were analyzed for correlation and consistency with activated partial thromboplastin time(APTT), fibrinogen(Fib), and platelet count(Plt), and the parameters tested when ECMO was established and 24 hours after ECMO operation were compared. 【Results】 The correlation coefficient between R values and APTT of hmTEG(r=0.441, P<0.05) was lower than that of ordinary TEG(r=0.547, P<0.05). The parameters α-Angle and K value of ordinary TEG were not correlated with Fib(P>0.05), while as for hmTEG, the correlation was 0.359(P<0.05) and -0.343(P<0.05), respectively. The correlation between MA value of hmTEG and Plt was 0.456(P<0.05), which was much lower than its correlation with Fib(r=0.715, P<0.05). APTT and hmTEG had moderate agreement in judging the anticoagulant effect of UFH(P<0.05). Plt at 24 hours after ECMO was significantly lower than that at establishment of ECMO(P<0.05). Fib, APTT and hmTEG parameters were not significantly different between the two groups(P>0.05). 【Conclusion】 The parameters of hmTEG can better reflect the real level of coagulation factors in patients receiving ECMO. The results of hmTEG and APTT are complementary to assess whether heparin in ECMO patients is overdosed, and hmTEG has unique advantages. Routine coagulation tests and TEG cannot replace each other, and the combination of them can achieve better anticoagulation and coagulation management.