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Objective To study the polymorphisms of short tandem repeat (STR) loci in intron 1, 24, 13 and 22 (STR 1,24, 13, 22) of factor Ⅷ (FⅧ) gene in Chinese population, and establish single tube multiple fluorescent PCR method for rapid diagnosis of haemophilina A(HA). Methods Four STRs from genomie DNA of 220 females without blood relationship were amplified in a single tube using quadri-fluorescence PCR. Capillary electrophoresis was analyzed in ABI PRISM 310 Genetic Analyzer. DNA sequencing was used to assay the number of dinueleotide repeats. Gene diagnosis were performed in 96 HA families. Results It was observed that 7, 9, 10 and 7 different alleles were found in STR1, 24, 13 and 22, respectively. The PIC (polymorphism information contents) were 0. 3789, 0. 4055, 0. 5239 and 0. 4713 in STR1,24, 13 and 22, respectively, and the HR (heterozygesity rate) were 34. 55% (76/220), 38. 18% (84/220), 49. 55% (109/220) and 43.64% (96/220). In 96 HA families, the diagnosis rate of STR1, 24, 13 and 22 were 38. 54% (37/96), 38. 54% (37/96), 54. 17% (52/96), 42. 71% (41/96), respectively. Whereas it achieved 79. 17% (76/96) when combining the four STR markers. Conciusion The single tube multiple fluorescent PCR of four STR loci is an effective, simple, quick method for linkage analysis and gene diagnosis of haemophilia A.
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Objective To test whether multiplex ligation-dependent probe amplification(MLPA)could be used for the prenatal detection of the most common aneuploidies of chromosomes 13,18,21,X,and Y.Methods 34 cases including 22 blood samples(12 with trisomy 21,1 with monosomy X,one male witll extra Y and 8 healthy persons),4 cord blood samples with Down syndrome and 8 amniotic fluid samples ( 1 with trisomy 21 and 7 normal fetuses)were recruited into this study.All samples were confirmed by karvotype analysis. DNA was extracted from blood and amniotic lysate was incubated with proteinase K.MLPA was used to determine the relative copy numbers.Results The resuhs were available within 48 h and were concordant with karyotype analysis in all but one case of amniotic fluid that was suggested to be triploid sample 69,XXY by MLPA or contaminated by maternal blood.This sample actually was found containing a number of red blood cells after centfifugation in test. In total,the concordance rate with clinical characteristics was 97.1%.The Ratio values of 13,18,21,X in normal samples were approaching 1.0 except chromosome Y having slightly higher variation in relative copy number.The difference of ratio means between the normal and trisomy 21 samples was statistically significant by one-way ANOVA(F=298.906.P=0.000).Conclusion Computer assisted MLPA with high sensitivity is a rapid,simple,automatic and reliable method for detection of common chromosomal aneuploidies.
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Objective To explore the relationship between MTHFR gene polymorphism and ~diabetic cardiovascular disease. Methods The MTHFR C677T and MTHFR A1298C gene polymorphisms were detectd by PCR-RFLP to compare MTHFR C677T and MTHFR A1298C genotype distribution and allele frequencies among three groups. Enrolled were 84 healthy individuals (NC) and 158 T2DM subjects who were classified into two groups, i.e. non-diabetes complications (NDC) and ~diabetic cardiovascular disease (DC). Results Patients with diabetic cardiovascular disease had higher frequencies of MTHFR C677T mutation genotype and T allele than control and non-diabetes complications subjects, and had lower folate levels (P0.05). A logistic regression analysis demonstrated that MTHFR C677T and age were the independent predictors of DC. Conclution There is a strong correlation between the polymorphism of MTHFR C677T and diabetic cardiovascular disease.
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<p><b>OBJECTIVE</b>In terms of the problems of now available dental machinable ceramics, a new type of calcium-mica glass-ceramic, PMC-I ceramic, was developed, and its machinability was compared with that of Vita MKII quantitatively. Moreover, the relationship between the strength and the machinability of PMC-I ceramic was studied.</p><p><b>METHODS</b>Samples of PMC-I ceramic were divided into four groups according to their nucleation procedures. 600-seconds drilling tests were conducted with high-speed steel tools (Phi = 2.3 mm) to measure the drilling depths of Vita MKII ceramic and PMC-I ceramic, while constant drilling speed of 600 rpm and constant axial load of 39.2 N were used. And the 3-point bending strength of the four groups of PMC-I ceramic were recorded.</p><p><b>RESULTS</b>Drilling depth of Vita MKII was 0.71 mm, while the depths of the four groups of PMC-I ceramic were 0.88 mm, 1.40 mm, 0.40 mm and 0.90 mm, respectively. Group B of PMC-I ceramic showed the largest depth of 1.40 mm and was statistically different from other groups and Vita MKII. And the strength of the four groups of PMC-I ceramic were 137.7, 210.2, 118.0 and 106.0 MPa, respectively.</p><p><b>CONCLUSION</b>The machinability of the new developed dental machinable ceramic of PMC-I could meet the need of the clinic.</p>
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Dental Porcelain , Reference Standards , Dental Stress Analysis , Hardness Tests , Tensile StrengthABSTRACT
Objective To discuss the enrichment method of fetal DNA from maternal plasma on size-fractionated separation.Methods Antecubital venous blood (5 ml from each pregnant woman) was collected in EDTA-containing tubes.DNA was extracted from plasma and white blood cells, separately.The DNA was fractionated by agarose gel electrophoresis.Three sections of