The Title Of Thesis The Relationship Of Hcy-related Gene CBS And CystainC Polymorphism With Ischemic Stroke

BackgroundIschemic cerebral vascular diseases(ICVD) is a group of a sudden onset disease of cerebral blood circulation disorders, To short-term or persistent focal cerebral ischemia, hypoxia, causing the softening and necrosis of brain tissue, Clinical manifested as focal neurological deficits, or associated with disturbance of consciousness. It mainly includes four types:1.Transient ischemic attack 2.Artery thrombosis cerebral infarction 3. Embolic cerebral infarction 4. Lacunar infarction. The definite etiopathogenisis of ICVD are as follow:vessel wall diseases (hypertensive arteriosclerosis, atherosclerosis), heart disease and hemodynamics, blood components and blood rheology changes, and various types of embolus, brain vascular compression, injury, spasm, etc. whereas hypertension, diabetes, heart disease, hyperlipidemia, smoking and other risk factors of stroke were more likely leads to occurrence of the ICVD.In recent years, the etiology of ICVD and pathogenesis research has made great progress, high-Hyperhomocysteinemia is considered as another new independent risk factor of ICVD, The main factor that lead to a higher Hcy levels is a deficiency of the metabolic related enzymes, coenzyme vitamin. There are more researchs about CBS (844ins68 and G919A), MTHFR (C677T and A1298C), MTRR (A66G), MS (D919G) and other metabolic enzymes, and we found that there may be some correlations between the genetic polymorphism and the ICVD, may be one of the pathogenesis of ICVD. CBS T27796C gene mutation may also lead to changes of plasma Hcy levels, but there are few relevant research about the relationship between the T27796C gene mutation and ICVD.In recent years, the cysteine protease inhibitor (Cystatin C, Cys C)attached more attention by people,and trere are a great number of research about the relationship between Cys C and ICVD. Olsson T's research showed that,the number of cerebral infarction are more likely to be found in the Cys C gene knockout mouse model, it may be due to Cys C on the diminished role of protease inhibitors in Cys C gene knockout mice, so Cys C may be a protective factor of cerebral infarction. Atherosclerosis (AS) of this chronic inflammatory process has been considered as an important risk factor of ICVD. Research has found that the lack of Cys C may be one of the pathogenesis of AS, thereby indirectly promote the occurrence of ICVD. There are studies that Cys C can cause lesions of the protein amyloid deposition on blood vessel wall, consider Cys C may be related to cerebral infarction. Foreign research shows that elevated Cys C levels has direct relevance with ischemic and hemorrhagic stroke, Cys C is a risk of cerebrovascular events and a mortality predictors. Whether there is a relationship between Cys C gene Rs2897119 (-177C/T) polymorphism, blood hcy levels and ICVD, such research is still few.In this study, to be adopted case-control study to explore the relationships between the T27796C and Cys C Rs2897119(-177C/T) with ICVD, this gene locus polymorphism analysis using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) Technology, in order to further explore the relationships of the two gene locus with occurrence of ICVD.ObjectTo investigate the frequencies of CBS T27796C, Cys C Rs2897119 (-177C/T) gene polymorphisms, the relationships between gene polymorphisms and ICVD which came from Han nationality of He Nan province in China.Methods1. Study object:A case-control research was performed on 185 ICVD patients and 111 collator. All subjects came from Han nationality of He Nan province in China. All patients fulfilled the diagnostic criteria of ICVD proposed by the fourth nationwide cerebrovascular diseases academic conference, and final diagnosis by head CT and (or) MRI.185 ICVD patients and 111 collator were randomly selected from outpatients or inpatients in Neurology Department of the first Affiliated Hospital of Zhengzhou University from December 2008 to April 2009. None of the control subjects had a recorded personal or familial history of ICVD and clinical vessel incident occurred. The age and sex of the two groups were matched. Both groups were excluded cerebral hemorrhage, subarachnoid hemorrhage and other hemorrhagic cerebrovascular disease, non-Parkinson's disease, dementia history, no thyroid disease, no liver and kidney disease, without taking folic acid, vitamin B12, Madopar and other drug history.2. Peripheral venous blood samples were collected and anticoagulationed. with EDTA, saved into the refrigerator (4℃) and the whole genomic DNA was extracted from white blood cells.3. PCR was used to amplify the aim genetic sequence.The PCR products were separated on 2% agarose gel stained with 0.5% ethidium bromide.4. The PCR products were analyzed by restriction fragment length polymorphi-sm (RFLP) technology.5. All statistical analyses were performed using SPSS 16.0 statistical analysis software. Genotype distribution underwent the Hardy-Weinberg (HW) equilibrium calculation; the two groups are presentative of the overall population. Theχ2 test was used to compare genotype and allelic frequencies between the ICVD and control populations, significant level isα=0.05.Results1. There were significant difference of plasm tHcy levels existed in ICVD group compared with the control group (t=6.532, P<0.05).2. There were significant difference of genotype of CBS T27796C existed in ICVD group compared with the control group (P<0.05), and there were also several significant difference of allelic frequency of CBS T27796C existed in ICVD group compared with the control group (P<0.05). 3. There were not significant difference of genotype and allelic frequency of CBS T27796C existed in ICVD group compared with the control group (P>0.05).4. There were not significant difference of genotype and allelic frequency of CystainC Rs2897119 (-177C/T) existed in ICVD group compared with the control group (P>0.05).Conclusions1. Hyperhomocysteinemia is a significance influential factor about ICVD occurred.2. The polymorphisms loci may be located in CBS T27796C and CystainC Rs2897119, including T/T, C/C homozygote, C/T heterozygote.3. CBS T27796C polymorphism may be associated with ICVD genetic susceptibility, to a certain extent can change the incidence of ICVD risk, so T27796C gene polymorphism may be a genetic factors about ICVD.4.CysC gene rs2897119 (-177C/T) polymorphism loci may not occur ICVD genetic factors.