鼠星形胶质细胞重编程为诱导多潜能干细胞的研究

目的研究星形胶质细胞重编程为诱导多潜能(iPS)干细胞的可行性,为进一步研究iPS细胞诱导技术奠定基础。方法用分别含Oct4,Sox2,Klf4和c-myc因子的4种逆转录病毒载体病毒颗粒感染星形胶质细胞,获得iPS细胞并进行鉴定。结果感染后第28天具有胚胎干细胞形态的克隆出现,诱导效率为(0.015±0.005)%,且碱性磷酸酶(AP)染色及SSEA-1和Oct4免疫荧光染色均显阳性。结论星形胶质细胞可以重编程为iPS细胞,进一步证明了iPS细胞诱导技术的普遍适用性。     

利用激光散斑成像技术观察尤瑞克林对脑梗死大鼠脑血流的影响

目的 利用激光散斑成像技术研究尤瑞克林对大鼠脑梗死后局部脑血流的影响.方法 成年雄性SD大鼠24只,线栓法制备大鼠永久性大脑中动脉梗死模型.激光散斑成像系统观测缺血半球皮质及大脑中动脉供血区血流,2,3,5-三苯基氯化四氮唑(TTC)染色法测定脑梗死体积,并进行神经功能评分.结果 皮质及大脑中动脉供血区血流在大剂量组第1天及第2天给药后均有明显改善,部分大脑皮质血管增粗,血流速度加快,小剂量组及生理盐水组无明显变化,脑缺血48 h后,大、小剂量尤瑞克林组及生理盐水组的梗死体积分别为10.14%±3.02%,25.99%±3.90%,27.10%±3.32%,大剂量组与生理盐水组比较差异有统计学意义(F=61.14,P<0.01),小剂量组与生理盐水组比较差异无统计学意义.缺血后4 h,大剂量组神经功能损伤明显改善,小剂量组及生理盐水组无明显改变,36 h各组间的神经功能评分差异无统计学意义.结论 尤瑞克林可以减少大鼠局灶性脑缺血后梗死体积,延缓神经功能损伤,其作用可能与促进侧支循环的开放,增加大脑皮质和缺血区血流有关.     

尼莫地平对大面积脑梗死病人脑组织血供、NIHSS及Fugl-Meyer评分的影响

目的研究尼莫地平对大面积脑梗死病人的脑组织血供、美国国立卫生研究院卒中量表(NIHSS)及Fugl-Meyer评分的影响。方法选取我院2015年8月—2016年10月收治的大面积脑梗死病人96例,采用随机数字法将其分为对照组和观察组,每组48例。两组病人均接受脑梗死常规治疗,在此基础上,对照组病人接受依达拉奉治疗,观察组病人接受依达拉奉联合尼莫地平治疗,均连续治疗14d。比较两组病人的治疗总有效率和不良反应发生率,同时比较治疗前后血气指标、NIHSS及Fugl-Meyer评分。结果观察组治疗总有效率(91.67%)明显高于对照组(72.92%),差异有统计学意义(χ2=5.79,P=0.01);治疗后,观察组的氧合血红蛋白(HbO2)、血氧饱和度(SpO2)、总血红蛋白(HbT)明显高于对照组(P0.01),NIHSS评分明显低于对照组(t=4.14,P=0.00),Fugl-Meyer评分明显高于对照组(t=-7.93,P=0.00),而还原血红蛋白(Hb)比较差异无统计学意义(P0.05);两组病人的不良反应发生率比较差异无统计学意义(P0.05)。结论尼莫地平治疗大面积脑梗死的临床疗效显著,能够明显改善病人脑组织血流灌注,促进神经功能康复。     

Proliferation and differentiation of neural stem cells co-cultured with cerebral microvascular endothelial cells after oxygen-glucose deprivation

Various stem cells, including neural stem cells (NSCs), have been extensively studied in stroke models, but how to increase neuronal differentiation rate of NSCs remains unresolved, particu- larly in a damaged environment. The purpose of this study was to investigate the effects of cerebral mi- crovascular endothelial cells (CMECs) on the neurogenesis of NSCs with or without oxygen-glucose deprivation (OGD). The NSCs acquired from primary culture were immunostained to prove cell purity. Survival and proliferation of NSCs were determined after the co-culture with CMECs for 7 days. After removing the CMECs, NSCs were randomly divided into two groups as follows:OGD and non-OGD groups. Both groups were maintained in differentiation culture for 4 days to evaluate the differentiation rate. Mouse embryo fibroblast (MEF) cells co-cultured with NSCs served as control group. NSCs co-cultured with CMECs had an increase in size (on the 7th day:89.80±26.12μm vs. 73.08±15.01 μm, P<0.001) (n=12) and number [on the 7th day:6.33±5.61/high power objective (HP) vs. 2.23±1.61/HP, P<0.001] (n=12) as compared with those co-cultured with MEF cells. After further differentiation culture for 4 days, NSCs co-cultured with CMECs had an increase in neuronal differentiation rate in OGD and non-OGD groups, but not in the control group (15.16% and 16.07% vs. 8.81%; both P<0.001) (n=6). This study provided evidence that OGD could not alter the effects of CMECs in promoting the neuronal differentiation potential of NSCs. These findings may have important implications for the development of new cell therapies for cerebral vascular diseases.