目的探讨脑卒中患者血清同型半胱氨酸(homocysteine,Hcy)及神经元特异性烯醇化酶(neuron specific enolase,NSE)对其癫痫发作的影响。方法研究对象为2016年1月~2019年6月期间到保定市第四中心医院就诊的脑卒中患者,其中并发癫痫患者160...目的探讨脑卒中患者血清同型半胱氨酸(homocysteine,Hcy)及神经元特异性烯醇化酶(neuron specific enolase,NSE)对其癫痫发作的影响。方法研究对象为2016年1月~2019年6月期间到保定市第四中心医院就诊的脑卒中患者,其中并发癫痫患者160例,纳入癫痫组,随机抽取同期未并发癫痫患者160例,纳入对照组。采用ELISA(酶联免疫吸附试验)法检测所有患者血清中Hcy和NSE的表达水平。采用单因素考察癫痫发作影响因素;采用Pearson相关分析Hcy和NSE与癫痫患者卒中评分的关系;选取单因素中具有统计学差异的关键因素进行多因素logistic回归;绘制癫痫发作的ROC曲线,并计算曲线下面积(AUC)。结果癫痫组和对照组的年龄(≥80岁)、卒中类型(梗死/出血)、脑血管疾病家族史、高脂血症、同型半胱氨酸尿症、肺部感染、NIHSS评分(≥25分),BI评分≥55分)及病灶范围9个参数差异均有统计学意义(χ^(2)=5.472~18.315,均P<0.01);癫痫组的血清Hcy为51.34±7.48μmol/L,高于对照组的31.85±5.04μmol/L,经t检验具有显著差别(t=27.333,P<0.001);癫痫组的血清NSE为62.47±8.11ng/ml,高于对照组的35.40±5.43ng/ml,经t检验,差异有统计学意义(t=35.083,P<0.001)。Pearson相关分析显示,血清Hcy水平(r=0.314,P<0.001),NSE(r=0.352,P<0.001)与脑卒中NIHSS评分呈正相关,而血清Hcy水平(r=-0.338,P<0.001),NSE(r=-0.257,P<0.001)与BI评分呈负相关。Logistic回归分析可知,NIHSS评分(≥25分)(OR=3.781,95%CI 1.441~9.919),BI评分(<55分)(OR=3.546,95%CI 1.220~10.305)、大的病灶范围(OR=2.996,95%CI 1.137~7.738)、血清Hcy(OR=4.014,95%CI 1.590~10.134)及血清NSE(OR=4.325,95%CI 1.776~10.530)是影响脑卒中患者癫痫发作的独立危险因素。ROC曲线分析结果显示:血清Hcy的ROC曲线下面积为0.820,敏感度和特异度分别为79.2%和70.3%,血清NSE的ROC曲线下面积为0.873,预测的敏感度和特异度分别为83.7%和84.0%。结论脑卒中患者血清Hcy和NSE与其癫痫发展开更多
To develop a rabbit corneal endothelial (RCE) cell line, in vitro culture of RCE cells was initiated from Oryctolagus curiculus corneas and a novel RCE cell line was established in this study. To initiate the primary ...To develop a rabbit corneal endothelial (RCE) cell line, in vitro culture of RCE cells was initiated from Oryctolagus curiculus corneas and a novel RCE cell line was established in this study. To initiate the primary culture of RCE cells, corneas from rabbit eyes were sliced and attached into glutin-coated wells with endothelial cell surface down. After being cultured at a time-gradient interval from 48 to 6 h, the corneal slices were detached and reattached into new wells, respectively. Cells in the wells containing only a pure population of RCE cells were collected and cultured in 20% FBS-DMEM/F12 medium con- taining chondroitin sulfate, ocular extract, epidermal growth factor (EGF), basic fibroblast growth factor (bFGF), carboxymethyl-chitosan, N-acetylglucosamine hydrochloride, glucosamine hydrochloride, culture medium of rabbit corneal stromal cells and oxidation-degradation products of chondroitin sul- fate at 37℃, 5% CO2. The cultured RCE cells, in quadrangle and polygonal shapes, proliferated to con- fluence 3 weeks later. During the subsequent subculture, the shape of RCE cells changed gradually from polygonal to more fibroblastic. A novel RCE cell line, growing at a steady rate, with a population doubling time of 53.8 h, has been established and subcultured to passage 67. Chromosome analysis showed that the RCE cells exhibited chromosomal aneuploidy with the modal chromosome number of 44. The results of immuno-cytochemical staining with neuron specific enolase (NSE) confirmed that the RCE cells were in neuroectodermal origin. Combined with the results of vascular endothelial growth factor (VEGF) treatment and endothelial cell morphology recovery, it can be concluded that the cell line established here is an RCE cell line. This RCE cell line may serve as a useful tool in theoretical re- searches of mammalian corneal endothelial cells, and may also have potential application in artificial corneal endothelium development.展开更多
基金863 High-Tech Research and Development Program of China (Grant No. 2001AA625050)
文摘To develop a rabbit corneal endothelial (RCE) cell line, in vitro culture of RCE cells was initiated from Oryctolagus curiculus corneas and a novel RCE cell line was established in this study. To initiate the primary culture of RCE cells, corneas from rabbit eyes were sliced and attached into glutin-coated wells with endothelial cell surface down. After being cultured at a time-gradient interval from 48 to 6 h, the corneal slices were detached and reattached into new wells, respectively. Cells in the wells containing only a pure population of RCE cells were collected and cultured in 20% FBS-DMEM/F12 medium con- taining chondroitin sulfate, ocular extract, epidermal growth factor (EGF), basic fibroblast growth factor (bFGF), carboxymethyl-chitosan, N-acetylglucosamine hydrochloride, glucosamine hydrochloride, culture medium of rabbit corneal stromal cells and oxidation-degradation products of chondroitin sul- fate at 37℃, 5% CO2. The cultured RCE cells, in quadrangle and polygonal shapes, proliferated to con- fluence 3 weeks later. During the subsequent subculture, the shape of RCE cells changed gradually from polygonal to more fibroblastic. A novel RCE cell line, growing at a steady rate, with a population doubling time of 53.8 h, has been established and subcultured to passage 67. Chromosome analysis showed that the RCE cells exhibited chromosomal aneuploidy with the modal chromosome number of 44. The results of immuno-cytochemical staining with neuron specific enolase (NSE) confirmed that the RCE cells were in neuroectodermal origin. Combined with the results of vascular endothelial growth factor (VEGF) treatment and endothelial cell morphology recovery, it can be concluded that the cell line established here is an RCE cell line. This RCE cell line may serve as a useful tool in theoretical re- searches of mammalian corneal endothelial cells, and may also have potential application in artificial corneal endothelium development.
