目的 探讨神经导航辅助神经内镜经鼻蝶入路切除垂体瘤的疗效。方法 将2014年10月—2018年4月我院接诊的20例垂体瘤患者纳入本研究,按照随机数字表法均分为观察组和对照组各10例,对照组患者行常规神经内镜下经鼻蝶入路垂体瘤切除术,观察组患者行神经导航辅助神经内镜下经鼻蝶入路垂体瘤切除术,比较两组患者手术及术后住院情况(包括手术时间、术中出血量及术后住院时间)、手术效果(包括显效率和总有效率)、手术前后血清内分泌指标变化情况(包括GH、PRL水平)以及术后并发症情况(即术后并发症发生率)。结果 观察组患者手术时间、术中出血量及术后住院时间均低于对照组,差异有统计学意义(P＜0.05);观察组显效率和总有效率均略高于对照组,但两组间手术效果并差异无统计学意义(P＞0.05);观察组患者治疗后血清生长激素(GH)、催乳素(PRL)水平均低于对照组,差异有统计学意义(P＜0.05);两组患者术后并发症发生率无统计学差异。结论 神经导航辅助神经内镜经鼻蝶入路切除垂体瘤疗效显著,手术时间短、术中出血少、术后恢复快,可明显改善患者内分泌指标,值得临床推荐。

Objective To evaluate the efficacy of resection for pituitary tumors through transsphenoidal approach with endoscopic neuronavigation assisted. Methods From October 2014 to April 2018, 10 patients with pituitary tumor were operated in our hospital with neuronavigation, which were set as the observation group. Meanwhile, 10 patients with pituitary tumor underwent surgical treating through transsphenoidal approach without neuronavigation were set as the control group. SPSS 19.0 were used for statistical analysis to compare the difference between the two groups, including the operation time, blood loss, hospitalization time, hormone level, clinical total efficiency, and complications in the two groups. Results The operation time, blood loss and hospitalization time of the observation group were less than that of the control group, and there was statistical significance (P<0.05). The efficiency rate and overall efficiency rate of the observation group were a little more than that of the control group, but there was no statistical significance (P＞0.05). The level of growth hormone (GH) and prolactin (PRL) of the observation group were less than that of the control group, and there was statistical significance (P<0.05). And there was no statistical significance between the incidence rate of postoperative complications of the two groups. Conclusion The efficacy of resection for pituitary tumors through transsphenoidal approach with endoscopic neuronavigation is significant, which may shorten the course, reduce the blood loss, quicken recovery of a patient from operation, and improve the hormone level. It is worthy to be recommended to clinical application.

目的 探索内源性神经干细胞在大鼠海马可溶性因子中的体外发育归宿及分化鉴定。方法 显微镜下分离Wistar大鼠海马组织放置于低温DMEM/12培养基,低温振荡2小时后高速离心(15000 g),获取实验所用海马组织可溶性因子。取材出生1天的Wistar乳鼠海马中的内源性神经干细胞(endogenous neural stem cells, ENSCs),将ENSCs分别于含海马可溶性因子终浓度为0(对照组)、50、100、200、400 μl/mL的无血清DMEM/F12培养基中培养6天并每日观察,使用免疫细胞化学、Western Blot印记技术比较各组ENSCs中Nestin、CD133的表达量;同时计量并比较各组ENSCs成球个数,以探索在模拟颅内微环境情况下,ENSCs发育、归宿及分化。进一步于最适宜的海马可溶性因子终浓度中分化神经球,对分化的细胞行神经元特异性蛋白入(如:β-tubullin III、MAP2)及胶质细胞特异性蛋白(如:GFAP、S100及p75 NGFR)免疫细胞化学检测。结果 大鼠ENSCs在培养基中呈单细胞漂浮生长,球形; ENSCs于海马可溶性因子各实验分组中培养第2天呈细胞球状态,对照组中无细胞球形成(与100 μl/mL组比较,P₁=0.00),100 μl/mL组与对照组比较有统计学意义(P₁=0.00<0.05);至第6天,在100 μl/mL组中的细胞球数量明显多于其余各组(P₁'=P₂'=P₃'=P₄'=0.00)。在免疫细胞化学检测中,100 μl/mL组中细胞球表达干细胞高亲和蛋白Nestin、CD133阳性,Western Blot免疫印迹检测其中Nestin、CD133蛋白高于对照组。进一步分化试验中,细胞球呈贴壁生长的单细胞状态、有突起伸出、长梭形,免疫细胞化学检测分化的细胞表达胶质细胞特异性蛋白GFAP、S100、p75NGFR阳性,但不表达神经元特异性蛋白β-tubullin III与MAP2。结论 大鼠ENSCs在终浓度为100 μl/mL的HSF作用下,可促进 ENSCs的增殖分裂;ENSCs在同样浓度下的HSF中可进一步分化为表达GFAP、S100、p75NGFR阳性的胶质样细胞;100 μl/mL的HSFS是ENSCs的一种生理性诱导剂或参与促进ENSCs增殖、分化及通过细胞替代或因子分泌等机制修复神经损伤。

Objective The aim of this study was to explore induction and differentiation of endogenous neural stem cells(ENSCs) in the hippocampus soluble factors(HSF) from the hippocampus of adult Wistar rats by mimicking an intracranial microenvironment. Methods After Wistar rats sacrificed, the hippocampus tissue was obtained in cold DMEM/F12. After centrigued and filtered, the HSF was stored at -20℃. The ENSCs was obtained from the hippocampus tissue of a neonate Wistar rat. Collected the tissue, digested and obtained the ENSCs. After we observed the morphology, the ENSCs were cultured in different concentration (0、50、100、200、400 μl/mL) of HSF for 6 days, and compared the expression of Nestin and CD133 by immunocytochemistry. Meanwhile,we compared the Nestin and CD133 protein by western blot. And then we explored the optimal concentration of HSF by the numbers of all groups on the second and sixth day. Furthermore, we did the differentiated experiment using the same concentration of HSF. Results The number of neurospheres in the 100 μg/mL group was significantly higher than those in the other groups on the 6th day. Immunofluorescence revealed that the neurospheres from ENSCs in the 100 μg/mL group more highly expressed nestin and CD133 than control. This result was confirmed by western blot analysis. The neurospheres can differentiate into glia-like cells in 100 μg/mL HSF and 1% FBS expressing GFAP, S100 and P75 NGFR by immunofluorescence. Conclusion These data indicated that HSF alone, mimicking a destination of ENSCs in vitro, could induce and differentiate neurospheres from ENSCs, as a new method to get NSCs and glia-like cells differentiated from ENCs to repair the diseases of center nervous system.