Combined polymorphisms in genes encoding the inflammasome components NLRP3 and CARD8 confer risk of Ischemic Stroke in men
Description
1. Research subjects The patients enrolled in this study, who were unrelated Chinese, included 234 cases with IS. The patients were hospitalized and treated at the Neurology Department, Nanxishan Hospital of Guangxi of China, from 2015 to 2017. One hundred fifteen controls, who visited the Medical Examination Center of the same hospital from 2015 to 2017, were also enrolled. 2. Risk factor assessment Demographic characteristics and vascular risk factors, including a history of smoking, hypertension, diabetes, dyslipidemia, and/or ischemic heart disease, were collected from the patients’ medical records or a structured questionnaire from controls. In this study, hypertension was defined as high blood pressure (BP; systolic BP ≥ 140 mm Hg or diastolic BP ≥ 90 mm Hg). Diabetes was defined as an elevated fasting plasma glucose level (≥126 mg/dl). Dyslipidemia was defined by a diagnosis of the disease. Ischemic heart disease was defined as a history of myocardial infarction, unstable angina, coronary intervention or bypass surgery. 3. Inclusion criteria Patients were enrolled if they met the following criteria: (a) diagnosis of stroke as defined by WHO (rapidly developing clinical symptoms and/or signs of focal cerebral dysfunction with no other apparent cause and symptoms lasting for more than 24h); (b) an age of 18 to 85 years old; (c) NCCT or MRI results consistent with IS; (d) a Chinese ethnicity (subjects and his/her ancestors [2 generations] were from mainland China); and(e) no history of trauma, brain tumor/metastases, known single-gene stroke disorder, central nervous system vasculitis, intracerebral hemorrhage, subarachnoid hemorrhage, transient ischemic attack, cardioembolic stroke or autoimmune/inflammation-related disorders. The controls met all the inclusion criteria, except that they were stroke-free as determined by the questionnaire for verifying stroke-free status (QVSS). 4. Genotyping Genomic DNA was extracted from EDTA-anticoagulated blood specimens using a standard method. The PCR primers were designed according to the gene sequences in GenBank.Genotyping was performed in a LightCycler 480 II System using the TaqMan SNP Genotyping Kit.LightCycler480 Software 1.5 was used to analyze the results. Ten percent of the samples were directly sequenced to confirm the genotyping results. 5. Assignment description: 5.1 Genotype assignment:Wild homozygous-1;Heterozygous-2;Mutant homozygous-3. 5.1.1 NLRP3rs10754558: CC-1; CG-2; GG-3. 5.1.2 CARD8rs2043211: AA-1; AT-2; TT-3. 5.2 Gender assignment: Male-1;Female-2 5.3 Group assignment: Control group-0; case group-1. 5.4 Native place: Crowd in southern China-1;Crowd in northern China-2. 5.5 Ethnic assignment:Han nationality-1;zhuang nationality-2;yao nationality-3. 5.6 Assignment of other information: 5.6.1 Nonsmoking-0;smoking-1 5.6.2 No hypertension-0;hypertension-1 5.6.3 No diabetes-0;diabetes-1 5.6.4 No coronary heart disease-0;coronary heart disease-1 5.6.5 No dyslipidemia-0;dyslipidemia-1