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OBJECTIVE@#To detect mutation of LBR gene in a pedigree affected with Pelger-Huёt anomaly (PHA) and to explore its clinical characteristics.@*METHODS@#Genomic DNA was extracted from the pedigree and healthy controls. The 14 exons of the LBR gene were subjected to PCR amplification and Sanger sequencing. Suspected mutations were verified in other family members and 100 healthy controls. Polyphen-2 and SIFT software were used to predict the effect of the mutation, and Swiss-model software was used to simulate the protein structure.@*RESULTS@#Three patients were found to carry a c.893G>A mutation in exon 8 of the LBR gene, which resulted in substitution of the 298th amino acid residue glycine by glutamic acid (p.Gly298Glu). The same mutation was not found in healthy family members and 100 healthy controls. The mutation was predicted to be damaging. Bioinformatic simulation showed the mutation has altered the 3D structure of the LBR protein.@*CONCLUSION@#The c.893G>A (p.Gly298Glu) mutation in the LBR gene probably underlies the PHA in this pedigree and has enriched the spectrum of LBR gene mutations.
Subject(s)
Humans , Case-Control Studies , DNA Mutational Analysis , Exons , Mutation , Pedigree , Pelger-Huet Anomaly , Genetics , Polymerase Chain Reaction , Receptors, Cytoplasmic and Nuclear , GeneticsABSTRACT
Objective@#To detect mutation of LBR gene in a pedigree affected with Pelger-Huёt anomaly (PHA) and to explore its clinical characteristics.@*Methods@#Genomic DNA was extracted from the pedigree and healthy controls. The 14 exons of the LBR gene were subjected to PCR amplification and Sanger sequencing. Suspected mutations were verified in other family members and 100 healthy controls. Polyphen-2 and SIFT software were used to predict the effect of the mutation, and Swiss-model software was used to simulate the protein structure.@*Results@#Three patients were found to carry a c. 893G>A mutation in exon 8 of the LBR gene, which resulted in substitution of the 298th amino acid residue glycine by glutamic acid (p.Gly298Glu). The same mutation was not found in healthy family members and 100 healthy controls. The mutation was predicted to be damaging. Bioinformatic simulation showed the mutation has altered the 3D structure of the LBR protein.@*Conclusion@#The c. 893G>A (p.Gly298Glu) mutation in the LBR gene probably underlies the PHA in this pedigree and has enriched the spectrum of LBR gene mutations.
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Objective To establish genotyping methods for vitamin K epoxide reductase complex subunit 1 (VKORC1)and cytochrome P450 2C9(CYP2C9)based on pyrosequencing technique to detection of warfarin metabolizing enzyme related gene polymorphisms.Methods A total of 50 peripheral blood samples from healthy adults were collected and the whole blood genomic DNA was extracted.A set of biotin-labeled amplifi-cation primers and sequencing primers were designed respectively for three SNP sites:VKORC1 -1639 G>A,CYP2C9 430C> T and CYP2C9 1075A>C.After PCR amplification of the samples,pyrophosphoric acid se-quencing was conducted.And then the signal peaks form were combined to analyze and determine each sample genotype.Genotyping results were verified by Sanger sequencing,and the consistency of the two sequencing methods was compared.Results Genotypes of the three SNPs can be clearly determined according to the ba-ses and height of the signal peaks.Among the 50 samples,there were 41 AA and nine AG for VKORC1 -1639G>A,accounting for 82% and 12% respectively,and there were 45 *1/*1,five *1/*3 for CYP2C9, accounting for 90% and 10% respectively,no CYP2C9*2 allele detected.Genotype results detected by pyrose-quencing and Sanger sequencing were consistent with each other.Conclusion In SNP genotyping,Pyrose-quencing has the advantages of convenience,time-saving,cheap with accurate and reliable results,which can quickly determine the genotypes of CYP2C9 and VKORC1.
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Objective To investigate the methods and internal quality control ( IQC ) leucorrhea routine examinationin clinical laboratories of medical institutions in Guizhou Province.Methods In 2009, 97 clinical laboratories were randomly selected for the first investigation.At the same time, staffs in theinvestigated laboratories were educated on the importance of IQC.The second investigation of the same items was carried out in 2011 inthe same laboratories.The results of the two investigations were analyzed byChi-square test.Results 2009 and 2011 numbers of laboratories thoseonly used normal saline suspension method for leucorrhea examination were 17and 16 (χ2 =0.037, P >0.05 ) respectively, used bothnormal saline and 10%KOH suspension methodswere 16and 2(χ2 =12.003,P<0.01), used staining method were 64and 79(χ2 =5.488,P<0.05), both used suspension and staining methods were 60and 73(χ2 =4.041, P<0.05), used normal salinesuspension method combined with Wright stain and Gram staining methods were3and 28(χ2 =23.996,P<0.01) respectively.Numbers of Laboratoriespracticing IQC were 2and 88in 2009 and 2011 respectivly(χ2 =153.293,P <0.01).Conclusions Currently, the most common used method for leucorrhea routine examination is suspension.Through the investigations and education, the quality ofleucorrhea routine examination was improved in Guizhou Province.
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Objective To investigate the factors affecting platelet transfusion efficiency.Methods A total of 102 cases of leuke-mia patients were recruited in the study,whose platelet count were measured before platelet transfusion and 1,24 h after platelet transfusion,then corrected count increment(CCI)values were calculated.By using CCI combined with clinical manifestations,the ef-ficacy of platelet transfusion were evaluated.The platelet antibody were detected before platelet transfusion.Depending on whether there were platelet antibodies,complications,the number of times of platelet transfusion,the types of platelet,patients were grouped and their platelet transfusion efficiency and CCI values were compared.Results The total effective rate of platelet transfusions were 71.6%(73/102 ).Invalid transfusion group had higher platelet antibody positive rate (17.2%)than effective transfusion group (2.7%),the difference were statistically significant(P <0.05).Among the groups of different transfusion times,the tansfusion effi-ciency was statistically different(P <0.05).With the increase of the number of times of platelet transfusion,the platelet transfusion efficiency decreased.Comparison between different types of platelets showed different platelet transfusion efficiency,which was sta-tistically significant(P <0.05).1 h and 24 h CCI value,platelet antibodies and whether patients with complications were related(P<0.05).1 h and 24 h CCI values were both associated with platelet antibodies and complications(P <0.05).Conclusion Platelet antibodies,complications,times of platelet transfusion and types of platelet transfusion are affecting factors of the transfusion effica-cy in patients with leukemia.
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Objective To establish an external quality assessment ( EQA) system of genitourinary tract secretions routine testing in Guizhou Province and improve the overall testing level.Methods From 2009 to 2011, more than 50 clinical laboratories in different grade hospitals from Guizhou Province were enrolled as participating units every year.EQA was carried out twice a year.Each time, five slides of high quality Wright′s or Gram stain smear of the genitourinary tract secretions or photographs obtained from these smears were selected to send to the participating laboratories for testing, and the feedback results from each laboratory were analyzed.The qualification was judged by the coincidence rate equal to or more than 80%. The average coincidence rates of each time and each year were statistically analyzed by Chi-squared test. Results From 2009 to 2011, the number of EQA participating units increased from 55 to 96, with an average return rate of >80%.Coincidence rates <80%of the 6 EQA results in the 3 years were as follow:four times for coccobacteria (73.7%,77.8%,61.1%,77.1%), twice for bacillus (75.6%,79.3%) and coccobacillus (64.3%,52.1%), once for infusorian (79.7%), epithelial cells (76.1%), neutropenia (75.7%) and cleanliness (71.3%).There were six batches of 30 quality assessment controls (accounting for 20.0%) in the six EQAs had the coincidence rate of <80%.Eleven items of 30 quality assessment controls with 1 to 15 batches were unqualified ( average coincidence rate of<80%) respectively.The item with the highest total average coincidence rate was suspected gonococcus (94.2%), and two items with the lowest total average coincidence rates were coccus and coccobacillus ( 77.0%, 75.2%, respectively ) . Conclusions This EQA program carried out within a certain range of clinical laboratories achieved good results:participating units increased significantly;the total score of all the items showed an obviously upward trend;the quality awareness of clinical lab technicians has enhanced to a certain extent.In this study, EQA system of genitourinary tract secretion routine testing were preliminarily established in Guizhou province, which provided a reference model of internal quality control ( IQC ) and EQA for clinical laboratories and higher authorities, and will be bound to have a positive impact on improvement of the overall level of genitourinary tract secretion routine testing.
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Objective To investigate the association between the apolipoprotein E (apoE) gene polymorphism and the dose for warfarin individual maintenance. Methods The genotypes of 249 patients with warfarin treatment in maintenance doses were determined by PCR/DHPLC assay. The doses for warfarin maintenance were compared among patients with different genotypes. Results In the total of 249 patients, the frequencies of 2/ε2, ε2/ε3, ε2/ε4, ε3/ε3, ε3/ε4, ε4/ε4 genotype were 1.20%, 15.66%, 1.80%, 72.29%, 9.24%, 0.80%, respectively; the allele frequencies of ε2, ε3, ε4 were 9.44%, 84.74%, 5.82%, respectively. The warfarin dose of group ε2 (ε2/ε2, ε2/ε3) was (3.24±1.36) mg/d, slightly higher than that of group ε3 (ε3/ε3, 2.91±1.14 mg/d) or group ε4 [ε4/ε4, ε3/ε4, (2.98±1.05) mg/d], but the difference of the warfarin doses among the 3 groups did not reach statistical significance (F=1.848,P＞0.05). Conclusion ApoE polymorphism may be not a major genetic factor that influences the individual dose for warfarin maintenance.
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0.05).Compared to cultivation whose positive rate was 33.33%,the rates were obvious higher(P