| Leukoariosis (LA) is a common neuroimaging change in old people, it's basic pathologic feature is white matter diffuse demyelination, is a clinical Syndromes caused by many factors, mainly presents as cognitive deficits, depression, lower limbs disability, and so on. The mechanisms for LA are not fully understood, but it's relationship with cerebral ischemia has been fixed. Atherosclerosis (AS) is an important factor of LA. Now with the development of Genetics, some scholars think genetic mutation and genetic polymorphism are independent important factors of LA. Apolipoprotein E (ApoE) plays an important role in lipoprotein metabolism, exon 4 of ApoE gene has three allele, namely ε2, ε3, ε4. Results of some researchs have showed that ε4 is a significant risk factor for AS, so maybe the existence and quantity of ε4 also have certain related to LA. In order to examine the hypothesis, we tested the ApoE genotype and blood fat of 58 patients with LA and their 1: 1 controls, analysed the correlation between ε4 allele and LA, Furthermore evaluated the association of LA with other vascular risk factors.Subjects and methods1:1 case-control study, the patient group and the control group were setted. LA patients and their sex, age and educational condition matched controls were recruited from Department of CT and Department of neurology in Huai he hospital affiliated toHenan University, all had spiral CT picture which photoed within 1 month and diagnosed by specified persons. LA patients were defined according to such radiological features on CT: one or more >5mm either patchy or diffuse hypedensity areas around periventricular or subcortices (centrum semiovale), the CT values in LA areas are lower 5-10Hu than those in normal white matter areas.excluding patients with multiple sclerosis, suprarenal gland white matter malnutrition, ect.The survey included physical finding, vascular risk factors, MMSE score and Hachinski ischaemia condition. Subjects were drawn 3ml blood before breakfast, 2 ml was used for separating serum to test blood fat by enzyme methods, another 1 ml was put into EDTA-K2 tube for extracting DNA.Test of ApoE genetic polymorphism by RELP-PCR method:(1)Extract: template DNA was extrated from subjects' fresh blood or frozen blood using Whole Blood DNA Purification System.(2)Amplify:Primers P1 5'-ACAGAATTCGCCCCGGCCTGGTACAC-3' P2 5'-TAAGCTTGGCACGGCTGTCCAAGGGA-3'All reaction system was 50μl, including lμg/20μl template DNA10μ1, 10mM/0.5ml dNTPs 5μl, 26.25pmol/μ1 P1 0.4μ1, 26.88pmol/μ1 P2 0.4μ1, 5U/μ1 TaqDNA polymerase 0.5μ1, PH 8.3 10% buffer 5μ1, 10% Dimethysulphoxidc (DMSO) 3μ1,25mM和MgCl23μ1.The reaction conditions were: heated at 95℃ for 5min, cycled 30 times at 95℃ for 1min, 60℃ for Imin, 70℃ for 1 min, followed by 70℃ for 7 min.(3)Cleave: the reaction system was 30μ1, including amplified products 20μl, Hhal endonuclease 5μl, 37℃ 4.5h.(4)Polyacrylamidedel electrophoresis, dyed by silver nitrate, then to determine the subjects' genotype.Risk factors analysis used single-factor analysis, multiple-factors conditional logistical regression. ApoE genotype and allele frequency analysis used frequencycounting method, x2 test. Measurement datas were recorded as x±s, t test, all tests were completed in SPSS10.0Results58 cases of LA and their 1: 1 controls were included in the study, both patient and control had same nation, sex. there were not different in age and education condition.56 cases in patient group concomitanted with stroke, 48.3% were lacunar infarction, 41.4% were cortical, subcortical infarction.Among 58 cases of LA, 19 cases (32.76%) were diagnosed as frontal lobe area LA, 4 cases (6.90%) as parietal or occipital lobe LA, 17 cases (29.31%) as around periventricular areas LA, 10 cases (17.24%) as semiovale area LA, 8 cases (13.79%) as two areas or more LA.The score of MMSE in LA group was lower than that in control group, P<0.001. the number of patients with cognitive deficits in LA group was h...
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