目的 研究靶向NKG2A抑制剂抗头颈部鳞状细胞癌（HNSCC）的作用。方法 应用GEO和TCGA数据库分析NKG2A及其配体HLA-E单细胞表达情况、与患者预后以及免疫微环境的相关性。构建HNSCC皮下抑制瘤模型,流式细胞技术检测化学治疗（化疗）对免疫检测点NKG2A表达的影响。动物实验验证NKG2A抑制剂以及NKG2A抑制剂联合多西他赛化疗的抗肿瘤作用。结果 NKG2A（KLRC1）主要表达在NK细胞,少量表达在T淋巴细胞。HNSCC肿瘤高表达NKG2A/HLA-E（P<0.01）,与患者不良预后密切相关;肿瘤微环境中NKG2A/HLA-E与多个免疫细胞浸润以及免疫检测点表达密切相关（P<0.01）。动物实验显示化疗能上调T、B淋巴细胞表达免疫检查点NKG2A的表达水平（P<0.01）;化疗的基础上联合NKG2A抑制剂能更有效地介导抗肿瘤作用（P=0.013）。结论 化疗基础上联合NKG2A抑制剂能更有效地介导抗肿瘤作用,为探索HNSCC临床新策略提供实验和理论基础。

Objective To investigate the anti-tumor effects of NKG2A inhibitor on head and neck squamous cell carcinoma（HNSCC）．Methods The single-cell expression of NKG2A ,its ligand HLA-E and their correlations with patient prognosis and immune microenvironment were analyzed in GEO and TCGA databases．The subcutaneous tumor model of HNSCC was constructed,and the effects of chemotherapy on the expression of NKG2A on T and B lymphocytes were detected by flow cytometry．Animal experiments were used to confirmed the anti-tumor effects of NKG2A inhibitor and NKG2A inhibitors combined with docetaxel．Results NKG2A（KLRC1）was mainly expressed in NK cells,and a small amount was expressed in T lymphocytes．The high expression of NKG2A/HLA-E in HNSCC tumors（P<0.01）were closely related to poor prognosis．NKG2A/HLA-E in tumor microenvironment were closely related to the infiltration of multiple immune cells and the expression of immune checkpoints（P<0.01）．Animal experiments showed that chemotherapy could up-regulate the expression of NKG2A in T and B lymphocytes（P<0.01）．Chemotherapy in combination with NKG2A inhibitor could mediate more effective antitumor effects in HNSCC（P=0.013）．Conclusions Combined with NKG2A inhibitor on the basis of chemotherapy can mediate more effective anti-tumor effects,and this study may provide experimental and theoretical basis for exploring new clinical strategies of HNSCC．

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 检测外周血循环肿瘤细胞(circulating tumor cell, CTC)在原发性肝癌患者中的表达情况,并探讨CTC动态变化及其相对于甲胎蛋白(Alpha fetoprotein AFP)对原发性癌患者术后复发转移的预测作用。方法 收集原发性肝癌患者134例,肝脏良性病变患者72例,检测外周血 CTC 数目,同时检测AFP的表达水平,分析 CTC 与 AFP 的相关性。然后在134名原发性肝癌患者中筛选出成功行肝癌根治术的患者,共86例,检测这86名患者术前、术后外周血CTC和AFP,分析CTC和AFP对原发性肝癌术后复发转移的评估价值。结果 原发性肝癌患者外周血CTC阳性率高于肝脏良性病变患者,差异有统计学意义(P＜0.05);原发性肝癌患者CTC水平与AFP水平、淋巴结转移、肿瘤结节多少有关,与年龄、性别、肿瘤直径、分化程度、肝硬化有无、TNM分期无关;原发性肝癌患者CTC和AFP生存分析显示,原发性肝癌根治术后早期复发转移与CTC和AFP密切相关;CTC较阳性对术后复发转移具有更好的诊断价值,二者联合对复发转移预测价值最高。结论 CTC可以做为一个比传统肿瘤标志物更好的对原发性肝癌术后复发转移进行监测的指标,与肿瘤标志物联合检测预测价值更高。

Objective To detect the expression of peripheral blood circulating tumor cells CTC in patients with primary liver cancer and to explore the dynamic changes of CTC and its predictive effect on postoperative recurrence and metastasis of primary cancer. Methods The number of CTC in peripheral blood was measured in 134 patients with primary liver cancer and 72 patients with benign liver disease, the expression of AFP was detected, and the correlation between CTC and AFP was analyzed. Then 86 patients with primary liver cancer were selected from 134 patients with primary liver cancer who underwent radical hepatectomy. The values of CTC and AFP in evaluating recurrence and metastasis of primary liver cancer before and after operation were analyzed by CTC and AFP, in peripheral blood of these 86 patients. Results The positive rates of CTC in peripheral blood of patients with primary liver cancer were higher than that of patients with benign liver disease(P< 0.05). The levels of CTC in patients with primary liver cancer were related to AFP level, lymph node metastasis and the number of tumor nodules, but not to age, sex, tumor diameter, differentiation degree, liver cirrhosis and TNM stage. The survival analysis of CTC and AFP in patients with primary liver cancer showed that the early recurrence and metastasis of primary liver cancer after radical resection were closely related to the positive rate of CTC and AFP, and the positive rate of CTC was more effective than that of AFP positive in the diagnosis of recurrence and metastasis after operation, and the combination of the two had the highest predictive value for recurrence and metastasis. Conclusion CTC may be used as a better index to monitor postoperative recurrence and metastasis of primary liver cancer than traditional tumor markers. The combined detection prediction value of tumor markers is higher.

目的 探讨自噬激活剂和自噬抑制剂分别对鼻咽癌细胞CNE2放疗敏感性的影响。方法 利用RNA干扰技术使atg5基因沉默,构建自噬抑制细胞模型后,与雷帕霉素、氯喹分别处理的两组细胞一起,每天以X射线5Gy照射细胞,连续8天观察各组细胞的生长状况,并设置对照组。以MTT法及克隆集落形成法检测其细胞活力,用流式细胞仪分析其细胞周期。结果 与对照组相比,其他三组细胞存活率、克隆形成率、照射后存活率均显著降低(P<0.05);细胞周期检测除对照组外其他三组细胞集中在G0/G1期,其他两个时期比G0/G1期相对较少。结论 自噬抑制剂与激活剂和atg5沉默均能为CNE2放疗增敏,然而自噬激活剂的增敏效果好于其他,为增敏放疗提供实验依据,开辟新的放疗增敏途径。

Objective This study aimed to investigate the autophagy activators and inhibitors effects in nasopharyngeal CNE2 cells radiotherapy sensitization. Methods Atg5 gene silencing by RNA interference technology, two groups of cell autophagy inhibition were built by rapamycin and chloroquine respectively. Then 5Gy x-ray irradiation of cells was taken every day, after 8 days in a row in each group of cell growth and setting a control group. The cell viability was clonaled colony formation by MTT method assay and cell cycle by flow cytometry analysis. Results The three cell group survival rate, colony-forming rate and survival after irradiation were significantly lower (P<0.05) than the control group. Detection of cell cycle in addition to control three other groups concentrated in the G0/G1 period.That of two other periods was relatively fewer than that of the G0/G1 period. Conclusion Autophagy inhibitors, activators and atg5 silence improved the radio-sensitization to CNE2. The autophagy activator group improving the sensitivity was better than the others.This study provided evidence to sensitive radiotherapy, explored a new promising radiosensitization ways.

目的 分析乳腺癌细胞中Snail与MTDH基因的作用,明确Snail是否通过结合于MTDH的启动子区域促进乳腺癌转移。方法 克隆、转染Snail基因至乳腺癌细胞,观察过表达Snail的乳腺癌细胞中MTDHmRNA及蛋白表达的变化;再使用免疫共沉淀法检测Snail与MTDH基因的共作用。结果 转染Snail基因进入乳腺癌MDA-MB-435细胞后,转染组、空白组和对照组中MTDHmRNA的表达水平分别为1.61±0.22、1.02±0.18、0.99±0.20,转染组高于空白组和对照组,差异有统计学意义(P<0.05),而后两组表达无差异(P>0.05);Westren blot检测结果显示,Snail可促进MTDH蛋白的表达;免疫共沉淀显示,Snail与MTDH在细胞内存在相互结合作用。结论 Snail在乳腺癌细胞中可通过结合于MTDH基因的启动子区域,促进MTDHmRNA转录及相关蛋白的表达,从而导致乳腺癌转移。

Objective To investigate the function of Snail to MTDH gene in breast cancer cells. Methods We observed the changement of MTDHmRNA and protein expression in breast cancer cell line MDA-MB-435 after transfected with Snail gene. Then, we used co-immunoprecipitation to determine the domain of Snail and MTDH binding in vitro. Results After transfected with Snail gene into MDA-MB-435 cell, the expression levels of MTDHmRNA in transfection group, blank group and control group were 1.61±0.22,1.02±0.18,0.99±0.20. The level of transfection group was significantly higher than the other groups(P< 0.05). Western blot showed that the expression of MTDH protein can be promoted by Snail. Co-immunoprecipitation showed that Snail and MTDH are binding interactions in breast cancer cell line MDA-MB-43. Conclusion Snail can promote transcription and expression of MTDH in breast cancer cells by binding to the promoter region of the MTDH gene resulting in metastasis of breast cancer.

       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．