Ibute to ACI phenotype. Association studies between polymorphisms of TAFI and stroke have been reported for many different origin populations, but with conflicting results [17-19]. Akatsu et al. found no statistical correlation between TAFI Ala147Thr and Thr325Ile polymorphisms and cerebral infarction [18]. Ladenvall et al. also did not find any association between genetic variants of TAFI and overall ischemic stroke, but they did find an increased risk between TAFI polymorphisms and stroke subtypes [20]. More recently, Kozian et al. found that the Ile/Ile genotype at PubMed ID:https://www.ncbi.nlm.nih.gov/pubmed/13485127 position 325 of TAFI is associated with the incidence of stroke and the age at onset of first stroke [19]. Furthermore, the incidence of stroke is higher in TAFI Ile325Ile patients with predisposing risk factors for thrombotic events, such as diabetes mellitus, myocardial infarction or hypertension [19]. These lines of evidence suggest a role for TAFI polymorphisms in the pathogenesis of stroke. However, the association between TAFI polymorphisms and ACI has not yet been determined. Therefore, the present study aims Methyl 2-((4-nitro-1h-pyrazol-1-yl)methyl)benzoate to evaluate the existence of an association between four polymorphisms (-2345 2G/1G, ?690 A/G, ?38 A/G and +1583 A/T) of TAFI and ACI in a case-controlled study of a Han Chinese population.gender, cerebrovascular disease risk factors and previous and ongoing medication or systemic therapy. A total of 184 matched controls (107 males and 77 females) were recruited from the Health Examination Center of the Affiliated Hospital of Guangdong Medical College during the same time period. These control patients were comparable to the ACI subjects in age, sex, and race. The same exclusion criteria were used as above. Written informed consent was obtained from each participant PubMed ID:https://www.ncbi.nlm.nih.gov/pubmed/10572343 prior to enrollment in the study. The study was approved by the ethics committee of Guangdong Medical College and was conducted according to the principles of the Declaration of Helsinki.Genotyping of TAFI polymorphismsGenomic DNA was extracted from peripheral blood using the EZ-10 Spin Column Whole Blood Genomic DNA Isolation Kit (Sangon Biotech? Shanghai, China) according to the manufacturer's instructions. Four TAFI SNPs (-2345 2G/1G, ?690 A/G, ?38 A/G and +1583 A/T) were selected according to previous findings [15]. Genotyping of the TAFI gene for the presence of ?38 A/G and +1583 A/T mutations was performed by polymerase chain reaction (PCR) and restriction fragment length polymorphism (RFLP); TAFI ?345 2G/1G and ?1690 A/G mutations were identified by allele-specific polymerase chain reaction (AS-PCR). The primers used are listed in Table 1. The PCR and RFLP analysis wereTable 1 Methods and primers used in genotyping of TAFI -438 A/G, +1583 A/T, -1690 A/G and -2345 2G/1G polymorphismsGene polymorphisms TAFI -438 A/G TAFI +1583 A/T TAFI -1690 A/G Analysis PCR-RFLP Primers Forward:ACCAATGGGAATGTAGG Reverse:CCTTGCCTCTGACTTTT PCR-RFLP Forward:CTTGGCATGTCATTAGG Reverse:TGCGGCATTTGTTGACAMethodsStudy populationAS-PCRCommon Forward 2,2,3,3-Tetrafluoropropyl N,N'-diethylcarbamimidothioate trifluoromethanesulfonate a:CACCTGTA GACTTTTGC Special Forward a1:TTAACTATTT TGACTGTTTA Special Forward a2:TTAACTATTT TGACTGTTTG Common Reverse b: CACTGAAG GAGAGAAAGThe study recruited 225 patients with ACI (145 males and 80 females) from the Department of Neurology at the Affiliated Hospital of Guangdong Medical College in China between 2011 and 2013. Patients' diagnoses were verified with either computed tomography (CT) or magnetic resonance imaging (MRI) and were cliniconeuropathologically confirm.