神经元特异性烯醇化酶(neuron-specific enolase, NSE)是糖酵解途径中一种重要的同工酶;特异性位于神经元和神经内分泌细胞中。小细胞肺癌(small cell lung cancer,SCLC)细胞浆内含有神经内分泌颗粒,具有神经内分泌分化的特征,为恶性程度高的神经内分泌系统肿瘤。因此,NSE是SCLC诊断中最敏感的肿瘤标志物,在SCLC的临床诊断、治疗、预后均有重要应用价值,科学合理联合检测肿瘤标记物,将能为临床诊疗工作提供有力的帮助。

目的 构建抑癌基因SEMA3B真核表达载体pcDNA3.1-SEMA3B,并检测其对肺癌A549细胞恶性生物学行为的影响。方法 应用PCR扩增SEMA3B全长cDNA片段,构建真核表达载体pcDNA3.1-SEMA3B。克隆PCR、双酶切法、基因测序验证过表达载体构建成功。将pcDNA3.1-SEMA3B真核表达载体和空载体pcDNA3.1分别转染入A549细胞中,应用qRT-PCR、Western blot检测SEMA3B mRNA、蛋白表达水平的变化;MTS法检测细胞增殖;流式细胞仪检测细胞凋亡、细胞周期;克隆形成实验检测细胞集落形成能力。结果 SEMA3B基因扩增片段与预测片段一致,克隆成功,且测序鉴定证实真核表达载体构建成功。转染pcDNA3.1-SEMA3B真核表达载体可上调SEMA3B mRNA、蛋白表达水平,且可抑制A549细胞的增殖,诱导凋细胞亡,细胞被阻滞在G1期,抑制细胞集落形成能力。结论 成功构建了SEMA3B基因真核表达载体,抑癌基因SEMA3B在肺癌恶性生物学进程中可能发挥重要作用。

Objective To construct the eukaryotic expression vector of the cancer suppressor gene, SEMA3B, and research the effects on malignant biological behavior of lung cancer A549 cells. Methods By reverse transcriptase-polymerase chain reaction (RT-PCR), the full length SEMA3B gene was amplified and then was inserted into pcDNA3.1. The recombinant plasmid pcDNA3.1-SEMA3B was confirmed correctly through double enzyme digestion and PCR identification, which was transfected into lung cancer A549 cells by lipid media transfection. The untransfected A549 and A549 transfected with pcDNA3.1 were used as controls. SMEA3B gene was detected by qRT-PCR and western blot. MTS assay, flow cytometry, and colony formation test were performed to evaluate the effect of overexpression of SEMA3B gene on A549 cell proliferation, apoptosis, cell cycle, and colony forming ability. Results The amplied fragment of SEMA3B gene by PCR was consistent with the anticipated result, the SEMA3B gene was cloned successfully. And the recombinant plasmid pcDNA3.1-SMEA3B was constructed successfully through gene sequence identification. After transfection of pcDNA3.1-SEMA3B, SEMA3B mRNA and protein expression levels were raised, and overexpression of SEMA3B gene in A549 cells significantly inhibited the proliferation of A549 cells, induced apoptotic cell death, blocked cell cycle in the G1 phase, and suppressed cell colony-forming ability. Conclusion The recombinant pcDNA3.1-SEMA3B is constructed successfully. SEMA3B gene can significantly inhibit the malignant biological behavior of lung cancer A549 cells.

       目的     探讨CT增强延迟扫描技术在非小细胞肺癌术前诊断中的应用价值。方法    对2021年5月—2024年5月商丘市第一人民医院收治的82例非小细胞肺癌手术治疗患者进行回顾性分析，将其分为观察组，另选取82例肺部良性肿瘤患者作为对照组，收集其术前CT增强延迟扫描结果，以术后病理诊断结果为金标准，分析CT增强延迟扫描技术在非小细胞肺癌术前诊断中的应用价值。并对比不同临床病理特征非小细胞肺癌患者CT增强延迟扫描的CT增强值，采用Spearman相关性分析法分析CT增强值与非小细胞肺癌病理特征的关系。结果  CT增强延迟扫描显示观察组患者分叶征（12.50% vs 53.57%）、内部空泡征数量（6.25% vs 39.29%）低于对照组（χ ² =26.560、24.680，P＜0.05），观察组患者边缘毛刺（56.25% vs 17.86%）、胸部凹陷征（59.38% vs 14.29%）、高于对照组（χ ² =43.330、64.600，P＜0.05）；82例非小细胞肺癌通过CT增强延迟扫描共确诊79例，CT增强延迟扫描诊断对非小细胞肺癌的准确率为96.34%（79/82），与病理诊断结果100.00%对比差异无统计学意义（χ ² =3.060，P=0.080）；82例非小细胞肺癌平均CT增强值为（39.14±7.31），不同性别、年龄、肿瘤最大直径、淋巴结浸润情况患者CT增强值对比差异无统计学意义（P＞0.05），不同病理类型［腺癌（43.75±7.15）vs 鳞癌（34.74±6.12）］、细胞分化程度［中、低分化（45.71±7.21）vs 高分化（32.81±5.11）］、临床分期［Ⅰ期（31.03±2.12）vs Ⅱ期（36.61±3.13）vs Ⅲa期（46.32±6.83）］患者、淋巴结转移［是（42.75±4.21）vs 否（35.77±8.13）］CT增强值对比差异有统计学意义（t/F=5.243、8.804、84.828、4.378，P＜0.05）；Spearman相关分析结果显示：病理类型、细胞分化程度、临床分期、淋巴结转移与非小细胞肺癌患者CT增强值呈正相关（r=0.431，P=0.021；r=0.511，P=0.009；r=0.586，P=0.005；r=0.579，P=0.008，P＜0.05）。结论  CT增强延迟扫描技术对非小细胞肺癌术前确诊具有重要价值，其诊断准确率与病理诊断并无显著差异，且可通过CT增强延迟扫描技术确定患者CT增强值，从而为非小细胞肺癌患者术后病理特征判断提供参考。

      Objective  To explore the application value of CT enhanced  delayed  scanning in  preoperative  diagnosis of non-small cell lung cancer（NSCLC）．Methods  A retrospective analysis was conducted on 82 patients with NSCLCwho underwent surgical treatment in a hospital from May 2021 to May 2024．They were included into  an  observation  group and another 82 patients with benign lung tumors were included in the control group．The  preoperative CT enhanced  delayed scanning results were collected，and the postoperative pathological diagnosis was used as the “gold standard” to analyze the application value of CT enhanced delayed scanning in the preoperative diagnosis of NSCLC．And the CT enhancement values of delayed CT scans in NSCLC patients with different clinical and pathological features were compared，and Spearman correlation analysis was used to analyze the relationship between CT enhancement values and pathological features of NSCLC．Results  CT enhanced delayed scanning showed that the number of lobular（12.50% vs 53.57%）and internal vacuolar signs（6.25% vs39.29%）in the observation group was significantly lower than that in the control group（χ ² =26.560，24.680，P＜0.05），while the edge spicules（56.25% vs 17.86%）and chest depression signs（59.38% vs 14.29%）in the observation group were significantly higher than that in the control group（χ ² =43.330，64.600，P＜0.05）．A total of 79 cases of 82 NSCLC were diagnosed by CT-enhanced delayed scan，and the accuracy of CT-enhanced delayed scan diagnosis for NSCLC was 96.34%（79/82），with no significant difference from the pathological diagnosis result of 100.00%（χ ² =3.060，P=0.080）．The average CT enhancement value of 82 NSCLC cases was（39.14±7.31）．There was no significant difference in CT enhancement values among patients of different genders，ages，maximum tumor diameter，and lymph node infiltration（P＞0.05）．Patients with different pathological types ［adenocarcinoma（43.75±7.15）vs squamous cell carcinoma（34.74±6.12）］，degree of cell differentiation ［moderate，and low differentiation（45.7±7.21）vs high differentiation（32.81±5.11）］，clinical stage ［I（31.03±2.12）vs II（36.61±3.13）vs IIIa（46.32±6.83）］ and lymph node metastasis ［yes（42.75±4.21），vs no（35.77±8.13）］ CT enhancement had significant difference（t/F=5.243，8.804，84.828，4.378，P＜0.05）．The Spearman correlation analysis results showed that pathological type，degree of cell differentiation，clinical stage，lymph node metastasis were positively correlated with CT enhancement values in NSCLC patients（r=0.431，P=0.021；r=0.511，P=0.009；r=0.586，P=0.005；r=0.579，P=0.008）．Conclusions  CT enhanced delayed scanning has important value in preoperative diagnosis of NSCLC．Its diagnostic accuracy is not significantly different from pathological diagnosis，and the CT enhanced value of patients can be determined through CT enhanced delayed scanning，providing reference for postoperative pathological feature judgment of NSCLC patients．

       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．

       目的 评价不同间变性淋巴瘤激酶（ALK）抑制剂联合安罗替尼治疗非小细胞肺癌（NSCLC）的疗效。方法 收集ALK突变阳性NSCLC患者的临床资料，筛选服用ALK抑制剂疗效不佳再加用安罗替尼的病例。根据不同的用药方案分为阿来替尼+安罗替尼，塞瑞替尼+安罗替尼和克唑替尼+安罗替尼三个组别。记录患者联合用药前最近一次的影像学检查结果，并以此为基线按Recist1.1评价疗效，以病情进展、患者死亡、停药、改变治疗方案为终点计算各组患者的无事件生存期（EFS），收集肿瘤标志物、血常规和肝功、心功能、肾功能生化检测等指标数据，统计分析患者联合用药前后各项指标的变化。结果 经筛选，共纳入49例患者的临床数据。阿来替尼+安罗替尼组有23例，疾病控制率（DCR）为86.96%；平均EFS为（10.8±3.6）个月，中位EFS为8.3个月；塞瑞替尼+安罗替尼组有14例，DCR为71.43%；平均EFS为（6.5±2.9）个月，中位EFS为5.6个月；克唑替尼+安罗替尼组有12列，DCR为66.67%；平均EFS为（7.7±3.2）个月，中位EFS为7.2个月。阿来替尼+安罗替尼组的平均EFS长于另外两组（P＜0.05）。各研究组肿瘤标志物仅有CyFra21-1在克唑替尼+安罗替尼组在联合用药后升高（P＜0.05），生化检测和血常规指标在用药前后差异无统计学意义（P＞0.05）。结论  ALK抑制剂与安罗替尼联用，疗效最好为阿来替尼，其次为塞瑞替尼，最后为克唑替尼。三种ALK抑制剂与安罗替尼联用后，均未导致心、肝、肾功能和血细胞损害。

      Objective  To evaluate the efficacy of different anaplastic lymphoma kinase（ALK）inhibitors combined with anlotinib in the treatment of non-small cell lung cancer（NSCLC）．Methods  Clinical data of drug resistant NSCLC patients with ALK positive mutation was collected who were treated with ALK inhibitors and anlotinib synchronously．According to different regimens，three groups were set，alectinib+anlotinib，ceritinib+anlotinib，and crizotinib+anlotinib．The latest imageological examination  results of the patient before the synchronous therapy was set as the baseline to evaluate the therapeutic effect according to Recist1.1．The event free survival（EFS）of each group was calculated with disease progression，patient death，treatment discontinuation and changing regimen as endpoints．Data of tumor markers，hematology test，liver function，cardiac function，renal function biochemical examination was collected and analyzed statistically before and after the combination therapy，with P＜0.05 as the statistically significant difference．Results  After screening，clinical data of 49 patients were collected．Twenty-three patients in the alectinib+anlotinib group，with a disease control rate（DCR） of 86.96%；mean EFS was（10.8±3.6）months，median EFS of 8.3 months；14 patients in the ceritinib+anlotinib group，with a DCR of 71.43%，mean EFS was（6.5±2.9）months，median EFS was 5.6 months；12 patients in the crizotinib+anlotinib group，with a DCR of 66.67%，mean EFS was（7.7±3.2）months，median EFS was 7.2 months．EFS of alectinib+anlotinib group was longer significantly than the other two groups（P＜0.05）．Only CyFra21-1，increased significantly after the combination of crizotinib and anlotinib（P＜0.05）．No statistically significant difference in biochemical test and hematology test before and after the treatment（P＞0.05）．Conclusions  The therapeutic effect of ALK inhibitors with anlotinib was ordered，alectinib being the most effective，followed by ceritinib and finally crizotinib．The combination of ALK inhibitors with anlotinib did not cause any abnormal results in the examination of heart，liver，kidney and blood cells．