Relationship between Ag-NORs and T Lymphocyte Subsets in Children with Epilepsy

Abstract

With the acceleration of the pace of life, the number of children with epilepsy is increasing. This study was to investigate the relationship between Ag-NORs and T lymphocyte subsets in children with epilepsy. In the morning, 3ml of elbow vein blood was collected from epileptic children on an empty stomach. After standing at room temperature for 0.3-1 hour, the serum was centrifuged (2000 rpm, 4 minutes). 3 ml of mixed venous blood was collected in EDTA-K2 vacuum container. The sample was taken out within 5 hours after staining. 30 μ l of ltcite reagent and 30 μ l of anticoagulant were put into TruCount tube and mixed evenly. Then flow cytometry was used for detection, and facscomp software was used for automatic inspection and calibration. In the same way, the automatic analysis software muitiset was used to obtain 20000 white blood cells, count 8000-10000 lymphocytes automatically, and calculate CD4 + T lymphocytes, CD8 + T lymphocytes and CD4 + / CD8 + ratio. Ag-NORs detection: under aseptic conditions, 0.5ml anticoagulant blood was added into the medium flask containing rpm-1640 and incubated in the incubator at 37 ºC for 72 hours. After mixing the cell suspension at room temperature, put it into a 10ml glass tube, heat it and dry it. Wait until the temperature of the water solution tank rises to 80-90 ºC, and then put aluminum plate on it. The staining was placed under the microscope on the stage, and the image was adjusted to make the image analysis program effective. The ratio of nuclear area to nuclear silver staining area of 30 lymphocytes was counted. This ratio reflects the content of Ag-NORs in the nucleolar forming region of T lymphocytes. The ratio of Ag-NORs area to nuclear area was 0.32 ± 0.03, 0.38 ± 0.03 and 0.46 ± 0.03, respectively. This study is helpful to provide guidance for the treatment of epilepsy in children.