目的 研究核磁共振（MR）引导的海马保护技术应用于小细胞肺癌全脑放射治疗（放疗）的效果。方法 对确定行全脑放疗的30例小细胞肺癌脑转移患者,行常规放疗CT定位后以定位体位行全头颅MR平扫,将计算机断层扫描（CT）和MR的T1加权像在Monaco 5.1计划系统上进行精准融合,勾画全脑放疗及海马区域,在海马区域三维方向上分别外扩5、15 mm作为海马与计划靶区之间的剂量跌落,每一例患者在Monaco 5.1计划系统上按照不保护海马组织以及外扩5、15 mm进行保护设计3个容积旋转调强技术（VMAT）放疗计划,观察海马组织的平均及最大放疗剂量。结果 增加保护海马组织之后,3个放疗计划的D100均≥95%,每例的3个放疗计划间D100比较差异无统计学意义（P>0.05）;设置外扩5、15 mm的剂量跌落区后,左、右海马的平均剂量、最大剂量均明显降低,而且3个放疗计划的海马平均剂量、最大剂量之间对比差异有统计学意义。结论 小细胞肺癌脑转移患者进行全脑放疗时,利用MR引导的海马保护技术并设置外扩15 mm的剂量跌落区,能够显著降低海马的剂量,达到保护目的。
Objective To explore the application of MR guided hippocampal avoidant whole brain radiotherapy（WBRT）for small cell lung cancer（SCLC）．Methods Thirty SCLC patients with brain metastases who underwent WBRT were enrdled．After routine CT localization was performed,and a head MR was performed in a the same position．T1 weighted images of MR and CT images were accurately fused on the Monaco 5.1 planning system．The entire brain tissue and hippocampus region were delineated． The dose drop areas between the hippocampus and the planned target area were expanded 5mm and 15mm in the three-dimensional direction of the hippocampus,respectively．Three volumetric modulated arc therapy（VMAT）radiotherapy plans were designed for each patient on the Monaco 5.1 planning system based on whether the hippocampal tissue was avoid．The average and maximum doses of hippocampal tissue were observed．Results After the avoidance of hippocampal tissue,the D100 of the three radiotherapy plans reached ≥95%,and there was no significant difference in D100 between the three radiotherapy plans in each case．After setting dose drop areas of 5mm and 15mm for external expansion,the average and maximum doses of the left and right hippocampus were significantly reduced,and there was a significant difference in the comparison between the average and maximum doses in the hippocampus of the three radiotherapy plans．Conclusions MR guided hippocampal avoidant technology and the setting of a 15 mm dose drop area can significantly reduce the dose to the hippocampus in patients with SCLC undergo whole brain radiotherapy．
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.
目的 研究核磁共振（MR）引导的海马保护技术应用于小细胞肺癌全脑放射治疗（放疗）的效果。方法 对确定行全脑放疗的30例小细胞肺癌脑转移患者，行常规放疗CT定位后以定位体位行全头颅MR平扫，将计算机断层扫描（CT）和MR的T1加权像在Monaco 5.1计划系统上进行精准融合，勾画全脑放疗及海马区域，在海马区域三维方向上分别外扩5、15 mm作为海马与计划靶区之间的剂量跌落，每一例患者在Monaco 5.1计划系统上按照不保护海马组织以及外扩5、15 mm进行保护设计3个容积旋转调强技术（VMAT）放疗计划，观察海马组织的平均及最大放疗剂量。结果 增加保护海马组织之后，3个放疗计划的D100均≥95%，每例的3个放疗计划间D100比较差异无统计学意义（P＞0.05）；设置外扩5、15 mm的剂量跌落区后，左、右海马的平均剂量、最大剂量均明显降低，而且3个放疗计划的海马平均剂量、最大剂量之间对比差异有统计学意义。结论 小细胞肺癌脑转移患者进行全脑放疗时，利用MR引导的海马保护技术并设置外扩15 mm的剂量跌落区，能够显著降低海马的剂量，达到保护目的。
Objective To explore the application of MR guided hippocampal avoidant whole brain radiotherapy（WBRT）for small cell lung cancer（SCLC）．Methods Thirty SCLC patients with brain metastases who underwent WBRT were enrdled．After routine CT localization was performed，and a head MR was performed in a the same position．T1 weighted images of MR and CT images were accurately fused on the Monaco 5.1 planning system．The entire brain tissue and hippocampus region were delineated．The dose drop areas between the hippocampusand the planned target area were expanded 5mm and 15mm in the three-dimensional direction of the hippocampus，respectively．Three volumetric modulated arc therapy（VMAT）radiotherapy plans were designed for each patient on the Monaco 5.1 planning system based on whether the hippocampal tissue was avoid．The average and maximum doses of hippocampal tissue were observed．Results After the avoidance of hippocampal tissue，the D100 of the three radiotherapy plans reached ≥95%，and there was no significant difference in D100 between the three radiotherapy plans in each case．After setting dose drop areas of 5mm and 15mm for external expansion，the average and maximum doses of the left and right hippocampus were significantly reduced，and there was a significant difference in the comparison between the average and maximum doses in the hippocampus of the three radiotherapy plans．Conclusions MR guided hippocampal avoidant technology and the setting of a 15 mm dose drop area can significantly reduce the dose to the hippocampus in patients with SCLC undergo whole brain radiotherapy．
