大鼠脊髓冲击伤模型制作方法的改良探索

目的:探索一种制作稳定性强、重复性好的脊髓损伤模型的简易方法。方法:应用简易重物坠落打击装置建立大鼠脊髓中度损伤组(5g×8cm)、重度损伤组(5g×16cm)模型和假手术组,每组大鼠均n=10。通过运动功能评分法(BBB)评分、苏木精-伊红染色观察脊髓损伤后两组大鼠功能及病理的变化特点,以评价该制作方法的可靠性。结果:BBB评分显示两组大鼠后肢运动功能均有不同程度的恢复,但中度损伤大鼠的恢复明显优于重度损伤大鼠(P<0.05)。组织病理学观察显示脊髓损伤后脊髓结构紊乱,有胶质瘢痕及空洞形成,重度损伤组脊髓空洞面积明显大于中度组(P<0.05)。结论:该方法制作的大鼠脊髓模型能将不同打击力度造成的损伤区分开,并且模型的行为学与病理学结果相吻合,说明此模型具有良好的稳定性、重复性和一致性,适合脊髓损伤实验研究的应用。     

老年缺血性脑卒中患者血清sFas和sFasL水平变化及临床意义

目的探讨老年缺血性脑卒中患者血清可溶性细胞凋亡因子(s Fas)及其配体(s Fas L)水平变化与病情发展的关系及临床意义。方法选取2011年6月至2013年6月来该院进行治疗的老年缺血性脑卒中患者63例作为观察组,采用双抗体夹心酶联免疫吸附试验(ELISA)法对患者不同病情发展阶段的血清s Fas和s Fas L水平进行测定,同时以20例老年健康体检者作为对照组。结果观察组血清s Fas和s Fas L水平显著高于对照组(P<0.05);患者在发病24 h后血清s Fas和s Fas L的表达开始升高,且在48 h达到高峰,并于1 w后开始下降,以后逐渐接近正常水平;患者病情越重,血清s Fas和s Fas L的表达越高,重度脑卒中患者血清s Fas和s Fas L的表达均高于中度和轻度患者;中度脑卒中患者均高于轻度患者(均P<0.05)。s Fas和s Fas L表达水平与脑卒中病情呈正相关(P<0.05)。结论 s Fas、s Fas L水平能反映老年缺血性脑卒中患者的早期病情发展,对临床病情判断及治疗方案的制定有一定的指导作用。     

降钙素原与血清和肽素及C-反应蛋白联合检测在脑出血肺部感染患者早期诊断中的价值和意义

目的探讨降钙素原、血清和肽素以及C-反应蛋白联合检测在脑出血并发肺部感染患者早期诊断中的临床价值。方法选取2016年1月-2017年2月医院就诊并确诊为脑出血患者98例,按是否合并肺部感染分为非感染组62例和感染组36例,采用酶联免疫吸附法(ELISA)测定两组患者血清和肽素及血清降钙素原水平;采取免疫荧光干式定量分析法,测定患者血清C-反应蛋白水平,比较两组患者发病后6h、24h、7d血清和肽素、降钙素原、C-反应蛋白水平,并比较不同指标的敏感度和特异度。结果感染组发病后6h、24h、7d血清和肽素、降钙素原、C-反应蛋白水平均高于非感染组(P0.05);将感染组发病后6h、24h、7d不同时间阶段血清和肽素、降钙素原、C-反应蛋白指标作为基础数据绘制ROC曲线,血清和肽素的敏感度、特异度分别为72.22%、83.33%,降钙素原的敏感度、特异度分别为88.89%、75.00%,C-反应蛋白敏感度、特异度分别为86.11%、77.78%,三个指标联合检测的敏感度、特异度分别为88.89%、94.44%。结论采取血清和肽素、降钙素原和C-反应蛋白联合检测的方式,可对脑出血并发肺部感染疾病进行早期诊断和鉴别,可明确指导肺部感染疾病选择合适的抗菌药物治疗。     

BMSC定向分化为神经干样细胞后移植改善脊髓损伤大鼠的神经功能

目的 研究骨髓间充质干细胞(BMSC)定向分化为神经干样细胞后进行移植,改善脊髓损伤大鼠神经功能的作用及其机制,探讨适宜的移植时间.方法 取培养至第3代的BMSC,将其定向分化为神经干样细胞,并采用免疫荧光染色法进行鉴定.移植前用5溴2脱氧尿嘧啶核苷(BrdU)标记细胞,移植时将其缓慢输注到损伤部位.实验分3组,移植Ⅰ组和移植Ⅱ组分别于脊髓损伤后1和2周进行移植,对照组操作同移植Ⅰ组,仅将移植用细胞悬液改为等量的生理盐水.移植后1~6周,对各组大鼠进行运动功能评分,采用荧光免疫化学染色检测移植细胞的存活、分化及神经纤维再生的情况.采用HE染色观察脊髓损伤区的病理学改变.结果 BMSC诱导培养3 d后,神经巢蛋白呈阳性表达,将诱导后的细胞球继续培养,可见细胞均有不同程度的神经丝蛋白(NF200)及胶质纤维酸性蛋白(GFAP)阳性表达.移植后两移植组大鼠的运动功能评分均显著高于对照组(P＜0.05),移植Ⅰ组尤为明显.2个移植组均有大量的5溴2脱氧尿嘧啶核苷(BrdU)和神经巢蛋白双阳性细胞及部分BrdU和NF200双阳性细胞填充于脊髓损伤区,同时可见明显的神经纤维再生,脊髓损伤处的空洞面积明显小于对照组(P＜0.05),而移植I组神经功能的改善较移植Ⅱ组更加明显.结论 BMSC可定向诱导、分化为神经干样细胞,将其移植后可有效改善脊髓损伤大鼠的神经功能,脊髓损伤1周后的移植效果优于损伤2周后移植.

Abstract：

Objective To study the effect and mechanism of the neurological function recovery in rats with spinal cord injury (SCI) rats after the transplantation of neural stem cells which are directly differentiated from bone marrow mesenchymal stem cells (BMSC), and to investigate the suitable engraftment time.Methods The BMSC at 3rd passage were differentiated into neural stem cells (NSC), and immunofluorescence staining was used to identify the NSC. The oriented-induced cells were labeled with Brdu 3 days before they were transplanted, and they were slowly injected into the injured site of the SCI rats. The SCI rats were randomly divided into group Ⅰ (transplantation at first week postinjury), group Ⅱ (transplantation at 2nd week postinjury) and control group (the operation was the same as group Ⅰ, but the cell suspension was replaced by the equal volume of normal saline). The BBB scores after transplantation were recorded. The distribution and differentiation of transplanted cells were observed by using immunofluorescence staining with antibodies against Brdu combined with Nestion and Brdu combined with NF200. NF200 immunofluorescence staining was used to show the regeneration of nerve fibers. The pathological changes of the injured site were observed by HE staining.Results The nestin expression was positive after the BMSC were differentiated for 3 days, and if the induced spherical cells were cultured continuously, the different levels of NF200 and GFAP were found. BBB scores in group Ⅰ and group Ⅱ were significantly higher than in control group (P<0.05), especially in group Ⅰ. The immunofluorescence showed that a large number of Brdu and Nestin double-positive cells and some Brdu and NF200 double-positive cells filled the injured site and linked the two sides of the injured area in group Ⅰ and group Ⅱ, and the lesion area of the spinal cord was reduced as compared with control group (P<0.05). More importantly, further reduction in lesion area and improvement in neurological function were observed in group Ⅰ.Conclusion The BMSC can be differentiated into NSC. The transplantation of the NSC could effectively promote the nerve function recovery after SCI, and the effect of transplantation at first week postinjury was better than at 2nd week postinjury.