1.Detection of HLA-B_(27) antigens by flow cytometry
Xianguo XU ; Xiaozhen HONG ; Ji HE
Chinese Journal of Blood Transfusion 1988;0(03):-
0.05) compared with the results of PCR ASP genotyping.The consistency between FCM and PCR ASP genotyping was 88% for positive results,100% for negative results,with a total of 95.9%.Conclusion Flow cytometric HLA B 27 typing is rapid and sensitive and can be used together with PCR technique for diagnosis of some diseases such as ankylosing spondylitis.
2.Molecular genetic analysis of Ael subgroup of the ABO blood group system
Xianguo XU ; Ji HE ; Xiaozhen HONG
Chinese Journal of Blood Transfusion 1988;0(02):-
Objective To understand the molecular genetic basis of Ael subgroup of ABO blood group system in the Han nationality.Method 2 Ael individuals were defined by standard blood group serological techniques,and genomic DNA was prepared for PCR SSP genotyping.Primers were designed and synthesized to amplify complete exon 6 and 7 including flanking intron sequence,and direct sequencing of gel purified PCR amplified fragments was performed using Bigdye Sequencing kit.Result A possibility of regarding the Ael allele as A2,B,O1 and O2 genes had been eliminated by the PCR SSP assay.According to the sequence analysis,Ael gene had 2 mutations of which one was a nucleotide substitution at position 532 in intron 5 (C to T),and the other was a single nucleotide(G) insertion at position 798 to 804 in exon 7 which alter the 86 amino acids sequence of the glycosyltransferase and furthermore extend the translated proteins by 37 amino acids compared with A1 allele.Conclusion The mutations of (789 804)G insertion and C(I 5/532)T substitution is the molecular genetic basis for Ael phenotype.
3.Research on molecular genetic basis for Jk(a-b-) phenotype
Faming ZHU ; Xianguo XU ; Xiaozhen HONG
Chinese Journal of Blood Transfusion 1988;0(04):-
Objective To investigate the molecular basis for Jk(a b ) phenotype.Methods Routine serologic testing for phenotype.Genomic DNA covering 4~11 exons and partial introns of JK gene was amplified by ploymerase chain reaction.The PCR products were excised and purified from agarose gels with a kit,then fragments were directly sequenced.Results G mutated to A in the 3'acceptor splice site of intron 5;A to G at 78 site from the 3'end of intron 3;C to T at 84 site from the 5'end of intron 8; A to G at 588 site of exons ( exon 7); G to A at 838 site of exons (exon 9).The splice site mutation (G→A) of intron 5 may cause the skipping of exon 6.Conclusion G to A mutation in the 3'acceptor splice site of intron 5 maybe one of the molecular basis for Jk(a-b-) phenotype
4.Identification the rare p phenotype in serological and molecular biological methods
Xiaozhen HONG ; Xianguo XU ; Faming ZHU
Chinese Journal of Blood Transfusion 1988;0(03):-
Objective To identify the p phenotype. Method P blood group system was identified using p phenotype cells,anti PP 1 P k antiserum,and direct DNA sequencing.Result and Conclusion Proband was typed as p, with rare anti PP 1 P k in the serum,family study suggested that inheritance was autosomal recessive.
5.Identification of the para-Bombay phenotype AB h m.
Xiaozhen HONG ; Xianguo XU ; Faming ZHU
Chinese Journal of Blood Transfusion 1988;0(01):-
Objective To identify para-Bombay phenotype AB h m. Method ABO and H phenotype were typed. Absorption and elution were performed. Saliva was tested by inhibitory reaction. Direct sequencing was performed and family study was done. Results Proband was typed as rare para-Bombay phenotype AB h mand anti-H was detected in his serum. Family study suggested that the inheritance was autosomal recessive. Conclusion Rare AB h m phenotype was identified and anti-H has been detected in his serum.
6.The clinical research about the relationship between serum levels of 8-hydroxy-deoxyguanosine (8-OHdG) of type 2 diabetes patients and its macrovascular complications
Xiaozhen YE ; Hong DU ; Jihai CHEN ; Huimei CHEN ; Jian WANG
Journal of Chinese Physician 2009;11(2):145-147
Objective To investigate the role of oxidative stress in type 2 diabetes patients with macrovascular complications. Methods Serum levels of 8-hydroxy-deoxygnan osine (8-OHdG) and hemoglobin Alc (HbAlc) in 32 cases of type 2 diabetic patients with macrovascular complications and 46 cases of patients without complications were determined. 8-OHdG was detected by enzyme linked immu-nosorbent assay (ELISA). Results The level of 8-OHdG (2.93±1.37) ng/mL in type 2 diabetic patients with macrovascular complica-tions was higher than those without complications, whose 8-OHdG level was (2.67±1.30)ng/mL, but the difference was not significant(P>0.05). Compared with all kinds of macrovascular complications in patients, the 8-OHdG levels were increased, but the difference were not significant (P>0.05). Compared with patients without complications, the duration and age of the patients with macrovascular compli-cations also increased significantly (P<0.05), but the HbAlc levels were not statistical difference between them (P>0.05). Spearman regression analysis showed that 8-OHdG level was positively correlated with the duration (r=0.33, P<0.05), while the age and HbAlc level had no obvious correlation (P>0.05). Conclusion The role of oxidative stress may not be the important reason in type 2 diabetes with macrovascular complications, but the duration and age of the patients may be closely related with the disease.
7.Nosocomial Infection Control in Local General Hospitals:Evaluation of Their Capabilities in Handling Public Health Emergencies
Jinlan HONG ; Huihui HE ; Xiaozhen QIU ; Jinfeng CHEN ; Chunmei CAI
Chinese Journal of Nosocomiology 2006;0(07):-
OBJECTIVE To evaluate the capabilities of nosocomial infection control in local general hospitals in handling public health emergencies,and to provide reliable data for future work.METHODS A random sampling questionnaire method was adopted to investigate how nosocomial infection control in local hospitals performed their functions and handled public health emergencies.RESULTS The 15 hospitals which were surveyed had all been equipped with computer network of directly reporting epidemic situations of infectious diseases.Four from 15 hospitals had full-time employees reporting epidemic situations,and 11 had part-time employees.Twelve hospitals established,according to standards,a department of infectious diseases or a department of pre-examination and sorting diagnosis.Seven hospitals did not have full-time staff of infection control till 2003.The rate of the staff's knowledge of nosocomial infection control was 73.7%.The medical wastes of the 15 hospitals were all disposed at the local medical waste disposal center.CONCLUSIONS Our city,in terms of nosocomial infection control,has acquired certain capabilities of handling public health emergencies.But the capabilities vary from hospital to hospital.Further improvement in some work is still needed.
8.Application of OMIM in medical genetics teaching and its reflection
Ya LI ; Kejian PAN ; Lan WANG ; Lang HE ; Hong LI ; Xiaozhen DAI ; Yuhan YANG
Chinese Journal of Medical Education Research 2013;(3):233-235
With the rapid development of medical genetics,online Medelian inheritance in man (OMIM) manifests a more and more important role in medical genetics teaching.Using the educational form combining ‘ classroom teaching,review writing and seminar’,‘ Query and use of OMIM ’was introduced into the education of medical genetics.Reality practice revealed that this educational practice maintained advanced and timely status of knowledge and deeply activated self-studying and independent thinking ability of students.
9.Detection of fetal RhCcEe genotype in maternal plasma
Ji HE ; Faming ZHU ; Xiaozhen HONG ; Jinhui LIU ; Shu CHEN ; Fei QIN ; Lixing YAN
Chinese Journal of Laboratory Medicine 2000;0(06):-
Objective To detect fetal RhCcEe genotype from fetal DNA in maternal plasma for noninvasive prenatal diagnosis.Methods DNA from maternal plasma sample was extracted by use of QIAamp DNA Kit. The existence of fetal DNA was confirmed by amplified fetal SRY gene. The fetal RhCcEe gene was amplified by polymerase chain reaction (PCR) from 30 pregnant maternal plasma. The results of fetal RhCcEe genotype were evaluated retrospectively by the serologic analysis of infant and pregnant woman RhCcEe phenotype.Results Among the 30 samples, 13 were the same phenotypes between mother and infant, 17 were different. When mother phenotypes were RhCC, cc, EE and ee homozygous, the deleted allele gene can be successfully amplified from mother plasma.Conclusion Noninvasive fetal RhCcEe genotyping is reliable. When the mother was homogyzous, genotyping the fetal CcEe alleles was very significant and useful for HDN (hemolysis disease of newborn) diagnosis and therapy.
10.Establishment and application of hnman platelet antigen genotyping with PCR sequencing-basod typing method
Xianguo XU ; Faming ZHU ; Ying LIU ; Xiaozhen HONG ; Kairong MA ; Xiaofei LAN ; Lixing YAN
Chinese Journal of Laboratory Medicine 2009;32(4):407-411
Objective To establish a PCR sequencing-based typing (PCR-SBT) method for simultaneous genotyping of human platelet antigen HPA-1 to HPA-16w.Methods All DNA polymorphism sites of HPA-1 to HPA-16w were obtained from the immuno polymorphism database.The specific primers were designed using Primer Premier 5.0 software to amplify nucleotide acid fragments encompassing each HPA polymorphism site.The primer sequence and PCR condition were optimized to obtain specific and single amplification product.The PCR product was purified and then sequenced to determine the HPA genotypes.Two standard DNA samples were detected using the HPA PCR-SBT method to examine the accuracy d this method.Sixteen reference samples (including 6 interference samples with HPA gene mutations) provided by 14th platelet immunology workshop of international society of blood transfusion (ISBT) in 2008 were also tested by this home-brew HPA PCR-SBT method.Results Total eleven pairs of primers were designed to amplify and sequence the sixteen HPA systems.The HPA genotypes of two standard samples were 1aa/2aa/3ab/4aa/5ab/6aa/7aa/8aa/9aa/10aa/11aa/12aa/13aa/14aa/15aa/16aa and 1aa/ 2aa/3aa/4aa/5aa/6aa/7aa/8aa/9aa/10aa/11aa/12aa/13aa/14aa/15aa/16aa,respectively.The 256 HPA genotypes of 16 reference samples were clear.128 genotypes among them were completely accordance with the results provided by ISBT report.Conclusions The PCR-SBT assay combining high-throughput DNA sequencer established in the study provides a simple,rapid and accurate method for HPA-1 to HPA-16w systems genotyping.The assay is suitable for routine clinical HPA genotyping and shows a broad prospect in further applications.