[摘 要] 目的：探究银杏内酯B（GKB）调控蛋白激酶R样内质网激酶（PERK）/转录激活子4（ATF4）/C/EBP同源蛋白（CHOP）信号通路对肝癌细胞增殖、迁移、凋亡和上皮间质转化（EMT）的影响。方法：将人肝癌细胞MHCC-97H随机分为对照组、GKB组、GSK2656157（PERK抑制剂）组和GKB + GSK2656157组，以GKB和GSK2656157分别干预后，采用MTT法和EdU染色检测各组细胞的增殖活性及增殖率，划痕愈合实验、流式细胞术分别检测各组细胞的迁移及凋亡水平，WB法检测各组细胞中EMT和PERK/ATF4/CHOP信号通路相关蛋白的表达水平。构建MHCC-97H细胞裸鼠移植瘤模型，以同法分组及药物干预后测定各组移植瘤体积，采用免疫组化、TUNEL染色分别检测各组肿瘤细胞增殖、凋亡水平，WB法检测各组移植瘤组织中EMT和PERK/ATF4/CHOP信号通路相关蛋白的表达水平。结果：与对照组比较，GKB组细胞活性、增殖率、迁移率、移植瘤体积、Ki-67阳性细胞率、MMP2、N-cadherin与MMP9蛋白表达均显著降低（均P < 0.05），细胞凋亡率、TUNEL阳性细胞率、p-PERK/PERK与E-cadherin、ATF4、CHOP蛋白表达均显著升高（均P < 0.05）；GSK2656157组各指标变化与GKB组相反（均P < 0.05）。与GKB组比较，GKB + GSK2656157组细胞活性、增殖率、迁移率、移植瘤体积、Ki-67阳性细胞率、MMP2、N-cadherin与MMP9蛋白表达均显著升高（均P < 0.05），细胞凋亡率、TUNEL阳性细胞率、p-PERK/PERK与E-cadherin、ATF4、CHOP蛋白表达均显著降低（均P < 0.05）。结论：GKB可通过激活PERK/ATF4/CHOP信号通路抑制肝癌MHCC-97H细胞增殖、迁移和EMT并促进其凋亡。

[摘 要] 目的：探讨嗜酸性粒细胞（Eos）和其他循环血细胞和炎症指标在预测小细胞肺癌（SCLC）免疫治疗疗效和免疫相关不良反应（irAE）中的价值。方法:回顾分析2013年8月至2023年7月期间海军军医大学第一附属医院呼吸科收治的410例SCLC患者临床信息；在化疗或免疫治疗前，以及治疗后的3个周期，分别检测患者全血细胞计数和细胞因子等指标；记录irAE的发生时间、类型、分级，以及随访情况。结果: 接受化疗联合免疫检查点抑制剂（ICI）治疗（简称联合治疗）的患者116例，其中一线联合治疗患者91例、后线联合治疗25例。联合组患者的总有效率（ORR）为44.8%，疾病控制率（DCR）为90.5%，单用化疗（单化）组患者的ORR为38.4%，DCR为85.0%。联合组中位PFS为8.9（7.2～10.5）个月，中位OS为17.7（13.9～21.5）个月。将联合组与单化组行倾向得分匹配（PSM）法配对，对比二组治疗后3周期的绝对嗜酸性粒细胞计数（AEC）水平、相对嗜酸性粒细胞计数（REC）水平；计算历次复查Eos水平与基线Eos水平的比值（AECT1/0、AECT2/0、AECT3/0、RECT1/0、RECT2/0、RECT3/0），联合组的AECT3/0和RECT3/0显著高于单化组。单因素分析表明，基线AEC和REC的升高与较好的治疗至失败时间（TTF）和OS显著相关（P < 0.05）；治疗后Eos水平与基线水平的比例（AECT3/0、RECT3/0）与较好的PFS和TTF显著相关（P < 0.05）；RECT3/0升高同样与OS改善显著相关（P < 0.05）。多因素分析提示，AECT3/0 > 0.41与较好的PFS和TTF显著相关（P < 0.05）；当RECT3/0 > 0.32时，与较好的PFS、TTF、OS均显著相关（P < 0.05）；RECT3/0 > 0.27仅与较好的TTF、OS显著相关（P < 0.05）。亚组分析发现，缓解组的RECT3/0显著高于非缓解组（P < 0.05）。一线应用ICI与二线/后线应用ICI患者的PFS、TTF、OS无统计学差异，但一线应用ICI时ORR（50% vs 25%，P < 0.05）和DCR（93.48% vs 79.17%，P < 0.05）显著优于二线/后线。116例联合治疗患者发生43例irAE（35.34%）, 最常见的为免疫相关性皮炎8.62%；Ⅲ级以上的irAE共17例（14.66%）；因irAE停药10例（8.62%），死亡2例;发生irAE的患者PFS较未发生irAE的患者显著延长（P < 0.05），而TTF和OS无统计学差异。发生irAE患者的RECT3较未发生irAE患者显著升高（P < 0.05），AECT3/0 > 0.29的患者irAE发生率显著增高（P < 0.05）。结论: Eos是SCLC接受ICI治疗的保护性因素，通过监测AECT3/0、RECT3/0，并结合患者的临床病理生理特征、细胞因子和炎症标志物水平进行综合评估，可有效预测SCLC患者的免疫治疗疗效和irAE的发生。

[摘 要] 目的：探讨颗粒酶M（GZMM）对肾透明细胞癌（ccRCC）细胞增殖、侵袭、迁移和血管生成的影响及相关分子机制。方法: 采用TCGA数据库和免疫组化分析GZMM在ccRCC组织的表达及其与临床病理特征的相关性。采用CCK-8、Transwell、划痕愈合及血管形成实验检测GZMM对ccRCC细胞增殖、侵袭、迁移和血管生成的影响，采用WB法检测GZMM对VEGF/ERK信号通路的影响。结果: TCGA数据库和免疫组化分析表明，ccRCC组织中GZMM的表达升高（P < 0.01），且与Fuhrman分级和淋巴结转移有关联（均P < 0.05）。GZMM高表达的患者预后不良（P < 0.05），且与FcerⅠ介导的MAPK激活有关联（P < 0.001）。在ccRCC细胞中，干扰GZMM降低ccRCC细胞的增殖、侵袭和迁移能力，且抑制ERK信号通路（均P < 0.05）；过表达GZMM促进ccRCC细胞的增殖、侵袭和迁移能力，且激活ERK信号通路（均P < 0.01）。在HUVEC中，分泌型GZMM促进HUVEC的增殖、迁移、小管和血管形成的能力，且激活VEGF/ERK信号通路（均P < 0.05）。此外，U0126抑制p-ERK、MMP2和MMP9的表达（均P < 0.05），但不影响VEGFA和VEGFR2的表达。结论：ccRCC组织中GZMM呈高表达，且与其Fuhrman分级和淋巴结转移有关联，GZMM通过激活VEGF/ERK信号通路促进ccRCC血管生成和侵袭。

[摘 要] γδT细胞是一类表达γδTCR异源二聚体的特殊固有免疫T细胞。过去，缺乏对其全面系统的基础研究，在其发育、分化、增殖、活化、效应和耗竭等所有环节仍有很多问题尚不清楚。然而，因为成熟的γδT细胞优势定植于皮肤、消化道、呼吸道、生殖道等肿瘤高发的黏膜组织，能以MHC非限制性的方式直接识别和杀伤多种肿瘤细胞，在肿瘤免疫治疗领域具有不可替代的优势，近年来其应用异军突起，发展迅速，也因此反过来促进了基础研究的深入，取得了一些亮眼的进展。本文对2023年γδT细胞在肿瘤免疫治疗领域的重大进展进行述评，主要集中在γδT细胞肿瘤抗原识别机制、肿瘤微环境中γδT细胞的功能调控、γδT细胞抗肿瘤细胞毒活性的机制、新型基于γδT细胞的肿瘤免疫治疗协同增效策略四个方面，以期推动γδT细胞在肿瘤免疫治疗领域的进一步发展，为临床γδT细胞应用协同增效的策略提供新的思路。

Objective: The present study was aimed to explore the role and distinctive mechanism of SPIDR, the key regulatory protein of homologous recombination pathway, in progression of small cell lung cancer (SCLC). Methods: 60 SCLC specimens and 44 normal lung tissues were collected from the patients undergoing tumor resection and bronchoscopic puncture in Shanghai Pulmonary Hospital Affiliated to Tongji University from January 2013 to January 2015. The expression of SPIDR in clinical samples and NCIH446 (SCLC cell line) and MRC-5 (normal cell line) were assayed by Real-time PCR. The role of SPIDR in SCLC was investigated in vivo and in vitro by the expression of SPIDR were artificially modified in NCI-H446. Results: Smoking was significantly associated with the occurrence of SCLC (P<0.01). The expression of SPIDR mRNAin SCLC tissues was lower than that of normal lung tissues (P <0.01), and the SPIDR transcriptional and translational levels of NCI-H446 cells were also lower than that of MRC-5.Although there is no significant changes of cell growth rate and susceptibility to cisplatin and etoposide in the NCI-H446 cells overexpressing SPIDR. However, the volume of xenograft tumors of overexpressed SPIDR group decreased by 58.99% (P<0.01) and 61.84% (P<0.01) than that of the original NCI-H446 cells and the NCI-H446 cells transfected with vector (pMSCV) and the average tumor mass decreased by 61.70% (P<0.01) and 70.25% (P<0.01) respectively. When the fetal bovine serum content in the medium was reduced to 3%, the growth rate of NCI-H446 cells overexpressing SPIDR was 22.33% (P<0.01) and 20.24% (P<0.05) lower than that of the original NCIH446 cells and control group, the similar results were obtained from the 1% serum concentration experiment as well. Conclusion: The expression of SPIDR, the key regulatory protein in the DNAdouble strand break homologous recombination repair pathway, was significantly suppressed in SCLC tissues, which markedly accelerated the growth of NCI-H446 cells in vivo and reduced the reliance of NCIH446 cells to the serum. The detailed mechanism is worthy of further investigation.

[Abstract] Objective: To study the effect and possible mechanism of TGF-β2 on the invasion of glioma stem cells (GSCs). Methods: Tumor tissues of 8 patients with glioblastoma multiforme, who underwent resection at Department of Neurosurgery of the FirstAffiliated Hospital of China Medical University duringApril 2016 toApril 2017, were collected. The primary culture of glioma cells were conducted with trypsin digestion. Partial primary glioma cells were seeded into serum-free DMEM/F12 culture medium containing EGF, bFGF and B27 to obtain suspension of tumor spheres. Immunoflurenscent staining and differentiation assay were used to detect whether the tumor spheres were GSCs. TGF-β2 secretion ability of GSCs was determined by ELISAassay.After transfection of TGF-β2 siRNA, the invasion ability of glioma stem cells was determined by Transwell assay. Western blotting was used to examine the effect of TGF-β2 on expression of matrix metalloproteinases (MMP) in glioma stem cells. Results: The suspended tumor spheres were proved to be GSCs by immunofluorescent staining and differentiation assay; the tumor spheres expressed the marker of GSCs（CD133）and had the ability to multi-differentiate (glia and neuronal cells). Compared with the primary glioma cells, Glioma stem cells exerted significantly improved TGF-β2 secretion ability ([74.13±3.63] vs [46.13±2.61] pg/ml, P<0.05); and TGF-β2 silencing significantly reduced the invasion ability of glioma stem cells ([105.71±8.69] vs [63.67±5.93], P<0.05) and inhibited MMP-2 and MMP-9 expressions. Conclusion: TGF-β2 can promote the invasiveness of glioma stem cells through MMP-2 and MMP-9 pathway.