ABSTRACT
<p><b>OBJECTIVE</b>To investigate the application value of the multiplex ligation-dependent probe amplification (MLPA) technique in diagnosis and prenatal diagnosis of chromosomes 13, 18, 21, X and Y aneuploidy.</p><p><b>METHODS</b>Forty-four cases including 30 peripheral blood samples, 10 fetal cord blood samples, and 4 amniotic fluid samples were collected in this study. DNA was isolated from the samples and detected by MLPA, followed by analyzing in ABI310 Genetic Analyzer. Analysis of copy number changes for chromosomes 13, 18, 21, X and Y was carried out with RH-MLPA-analysis software. The routine karyotype analyses were also done for all the samples.</p><p><b>RESULTS</b>Of 44 samples, the results of 42 by MLPA method was consistent with that by chromosome karyotyping. Only one case with trisomy 21 chimerism was failed to reach conclusion. In addition, one case of mark chromosome segment was identified as Y-chromosome segment by MLPA, while karyotyping failed to make judgment. The accurate rate of MLPA was 97.7% (43/44).</p><p><b>CONCLUSION</b>The MLPA technique can simultaneously detect dozens of different target sequences and their copy number changes in a single reaction. It showed high specificity, good reproducibility, was fast and high-throughput. The MLPA technique can be applied to diagnosis and prenatal diagnosis of the common chromosomal aneuploidy.</p>
Subject(s)
Female , Humans , Pregnancy , Amniotic Fluid , Chemistry , Aneuploidy , Chromosomes, Human, Pair 13 , DNA , Genetics , DNA Copy Number Variations , Down Syndrome , Diagnosis , Genetics , Fetal Blood , Chemistry , Nucleic Acid Amplification Techniques , Methods , Prenatal Diagnosis , Methods , Sensitivity and SpecificityABSTRACT
<p><b>OBJECTIVE</b>To detect the gene mutations of beta-myosin heavy chain gene (MYH7) in Chinese pedigrees with hypertrophic cardiomyopathy (HCM), and to analyze the correlation between the genotype and phenotype.</p><p><b>METHODS</b>Exons 3, 5, 7-9, 11-16 and 18-23 of the MYH7 gene were amplified with PCR in three Chinese pedigrees with HCM. The products were sequenced. Sequence alignment between the detected and the standard sequences was performed.</p><p><b>RESULTS</b>A missense mutation of Thr441Met in exon 14 was identified in a pedigree, which was not detected in the controls. Several synonymous mutations of MYH7 gene were detected in the three pedigrees.</p><p><b>CONCLUSION</b>The mutation of Thr441Met, located in the actin binding domain of the globular head, was first identified in Chinese. It probably caused HCM. HCM is a heterogeneous disease. Many factors are involved in the process of its occurrence and development.</p>
Subject(s)
Adult , Animals , Female , Humans , Male , Middle Aged , Amino Acid Sequence , Base Sequence , Cardiac Myosins , Cardiomyopathy, Hypertrophic , Genetics , DNA Mutational Analysis , Genotype , Molecular Sequence Data , Mutation , Myosin Heavy Chains , Chemistry , Genetics , Pedigree , PhenotypeABSTRACT
<p><b>OBJECTIVE</b>To improve the experimental method of DXS52 (St14) and apply it to genetic testing for hemophilia A (HA).</p><p><b>METHODS</b>PCR of DXS52 and agarose gel electrophoresis were performed for genetic testing in 61 non-inversion HA families. Linkage analysis of 7 loci within the FVIII gene including Bcl I, Hind III, Xba I, STR1, STR13, STR22 and STR24 were also carried out for the 61 families.</p><p><b>RESULTS</b>DXS52 can provide information in 43 out of 61 families and the diagnostic rate was 70.5%. Eight families can be diagnosed only by DXS52 locus, accounting for 13.1%. Two families were found to have recombination between DXS52 and FVIII.</p><p><b>CONCLUSION</b>The new experimental conditions can reach accurate and clear results in DXS52 genetic testing. This gene maker has high diagnostic rate, so it is an indispensable linkage analysis method in HA gene diagnosis. More caution should be paid when using the extragenic locus DXS52 to perform gene diagnosis because of its high recombinant rate with FVIII.</p>
Subject(s)
Female , Humans , Male , Chromosome Mapping , Methods , Chromosomes, Human, X , Factor VIII , Genetics , Genetic Linkage , Genetic Markers , Hemophilia A , Diagnosis , GeneticsABSTRACT
<p><b>OBJECTIVE</b>To detect gene mutations on beta-myosin heavy chain gene MYH7 in 3 Chinese families with hypertrophic cardiomyopathy (HCM), and to analyze the correlation between genotype and phenotype.</p><p><b>METHODS</b>A denaturing high-performance liquid chromatography (DHPLC) and sequencing mutation screening of the exons (exon3-23) coding for MYH7 gene were performed in 3 Chinese families with HCM.</p><p><b>RESULTS</b>In this study, we identified several mutations in MYH7. A mutation of Thr441Met previously reported in a patient with Laing distal myopathy was first identified in one Chinese pedigree.</p><p><b>CONCLUSION</b>This study illustrated the high frequency of mutation in MYH7 gene in Chinese HCM families. Different mutations and carriers of the MYH7 gene present phenotypic heterogeneity. Mutation screening and analysis in HCM family could therefore facilitate the early HCM diagnosis and would be helpful for the prediction, prevention and early treatment of HCM linked with MYH7 gene mutation.</p>
Subject(s)
Adolescent , Adult , Child , Female , Humans , Male , Middle Aged , Young Adult , Asian People , Genetics , Cardiac Myosins , Genetics , Cardiomyopathy, Hypertrophic, Familial , Genetics , Case-Control Studies , DNA Mutational Analysis , Exons , Genotype , Mutation , Myosin Heavy Chains , Genetics , Pedigree , PhenotypeABSTRACT
<p><b>OBJECTIVE</b>To research on the genetic polymorphism distributions of 15 short tandem repeat (STR) loci in Han race of North China and the genetic data of population genetics.</p><p><b>METHODS</b>The capillary electrophoresis and five-color fluorescent multi-amplifying were applied to detect the genotypes of 15 STR loci in 597 unrelated Chinese Han individuals.</p><p><b>RESULTS</b>No significant deviation from the Hardy-Weinberg Equilibrium was observed. High polymorphism was detected in the loci. Statistical analysis was carried out to obtain some parameters of forensic medicine. The heterozygosity of 15 loci was above 0.62. The values of discrimination power (DP) at 15 STRs ranged from 0.802 to 0.967. The values of excluding probability of paternity (EP) ranged from 0.320 to 0.697. The values of probability matching (Pm) ranged from 0.033 to 0.198. The fifteen loci showed an accumulated total discrimination power (TDP) more than 0.999999, a cumulative excluding probability of paternity (CEP) as 0.99999571, and total probability matching to be 8.93 x 10(-18).</p><p><b>CONCLUSION</b>The data indicated that detecting combined 15 STRs is sensitive and reliable, and can be used to forensic and individual identification cases in Chinese group.</p>
Subject(s)
Humans , Asian People , Genetics , China , Ethnology , Gene Frequency , Genetics, Population , Genotype , Microsatellite Repeats , Genetics , Polymorphism, Genetic , Tandem Repeat Sequences , GeneticsABSTRACT
To construct a safer and more efficient gene engineering Lactococcus Lactis for expressing phenylalaine ammonia lyase (PAL) which will be benefit for PKU therapy, pal cDNA of Parsly and synthesized sequence based on Lactococcus Lactis bias codons were recombined into two Lactococcus Lactis NICE systems. The activities of the expressed PAL were detected, and the effect of Lactococcus Lactis bias codons on the expression of exterior protein was analyzed. The results showed that the expression level of PAL was increased by using Lactococcus Lactis bias codons in both Lactococcus Lactis NICE systems. Through which several safer andmore efficient strains of the gene engineering Lactococcus Lactis were obtained.
Subject(s)
Cloning, Molecular , Codon , Genetics , Genetic Vectors , Genetics , Lactococcus lactis , Genetics , Metabolism , Phenylalanine Ammonia-Lyase , Genetics , Recombinant Proteins , Metabolism , Transformation, BacterialABSTRACT
<p><b>OBJECTIVE</b>To study the strengthening of fluorophlogopite-based dental machinable ceramic through developing a new type of calcium-mica-based ceramic.</p><p><b>METHODS</b>Based on the analysis of the crystal structure of the fluorophlogopite ceramic of Dicor MGC, the structure of a new type of calcium-mica-based ceramic was designed and the corresponding composition of the new material was experimented. And the new glass-ceramic was obtained through the treatment of glass preparation and nucleation. Then crystal content of the glass ceramic was analyzed by X-ray diffraction analysis (XRD) and the 3-point bending strength of the new ceramic was recorded.</p><p><b>RESULTS</b>A new type of calcium-mica-based glass ceramic was developed and had the 3-point bending strength of (210.2 +/- 14.7) MPa. Compared with the strength of fluorophlogopite-based dental machinable ceramic, such as Dicor MGC, which was reported in the range of 150 approximately 180 MPa within inclusive studies, the higher strength of the new ceramic was recorded.</p><p><b>CONCLUSIONS</b>The fluorophlogopite-based dental machinable ceramic could be reinforced through internal strengthening.</p>