B7-H4及Fas、FasL与宫颈癌免疫逃逸机制的关系

目的:检测免疫共刺激分子B7-H4及Fas、FasL在不同宫颈组织中的表达，研究三者的异常表达与宫颈癌免疫逃逸机制的关系。方法:选择陕西省肿瘤医院病理科2014-2018年的石蜡包埋标本162例，采用免疫组织化学方法检测20例正常宫颈、30例宫颈上皮内瘤变(CIN)和112例I期-IV期宫颈癌组织中B7-H4及Fas、FasL的表达水平。结果:B7-H4在正常宫颈组、CIN组、宫颈癌组的阳性表达率分别为0、20.0%、47.32%，两两相比，具有显著性差异（P<0.05）。在112例宫颈癌病例中FIGO临床分期I期至IV期各期别宫颈癌组织B7-H4的阳表达率分别为16.67%、43.18%、70.83%、85.71%，各组间相比差异具有统计学意义。正常对照组Fas的阳性表达率是85.00％，CIN组43.33％，宫颈癌组41.96％，正常组与CIN、正常组与宫颈癌组相比差异均具有统计学意义（P<0.05），CIN组与宫颈癌组相比，无明显差异（P>0.05）。FasL的阳性表达率在正常组为20.00%，CIN组为50.00%，宫颈癌组为56.25%，正常组与CIN、宫颈癌组比较，差异具有统计学意义(P<0.05)，CIN组与宫颈癌组比较无显著差异;B7-H4阳性宫颈癌组Fas阳性表达率28.30%明显较B7-H4阴性宫颈癌组Fas阳性表达率54.24%低，差异具有统计学意义（P<0.05）；而B7-H4阳性宫颈癌组FasL阳性表达率79.25%明显高于B7-H4阴性宫颈癌组FasL阳性表达率35.59%，差异具有统计学意义（P<0.05）。结论:负性免疫共刺激分子B7-H4和Fas、FasL凋亡蛋白在宫颈癌的发生、进展中起到了重要作用，其协同作用可能为宫颈癌细胞逃逸机体免疫的机制之一。     

N-乙酰-5-羟色胺对视网膜缺血-再灌注损伤大鼠视网膜 Fas、FasL 蛋白表达的影响

目的　探讨N-乙酰-5-羟色胺（N-acetylserotonin，NAS）对视网膜缺血-再灌注损伤（retina ischemia-reperfusion injury，RIRI）大鼠视网膜Fas、FasL蛋白表达的影响。方法　取健康成年Sprague Dawley大鼠54只，将大鼠随机分为正常组（6只）、RIRI组（24只）与NAS组（24只）；采用高眼压法建立大鼠RIRI模型，依据造模后不同时间点将RIRI组与NAS组大鼠又分为6 h、12 h、24 h及72 h四个亚组。NAS组于造模前30 min腹腔注射NAS（5 mg·kg^-1），RIRI组腹腔注射等剂量的生理盐水。通过HE染色在光学显微镜下观察各组大鼠视网膜形态学变化，并记录各组大鼠视网膜厚度及视网膜神经节细胞数，采用免疫组织化学染色法检测NAS对RIRI大鼠视网膜Fas、FasL蛋白表达的影响。结果　HE染色显示，正常组大鼠视网膜各层细胞分界清晰，形态正常，神经细胞排列整齐；RIRI组大鼠再灌注后6 h视网膜各层出现水肿，以神经节细胞层及内核层较显著，神经节细胞数较正常组减少；随后视网膜水肿进一步加重，神经节细胞继续减少；NAS组大鼠在再灌注后6 h、12 h、24 h 视网膜水肿程度较 RIRI组轻，NAS组在再灌注后72 h视网膜厚度较 RIRI组厚，NAS组各时间点神经节细胞数均较 RIRI组多，差异均有统计学意义（均为P＜0.05）。免疫组织化学染色显示，正常组几乎未见 Fas⁺细胞。再灌注后6 h，RIRI组视网膜神经节细胞及内核层开始出现少量 Fas⁺细胞；再灌注后12 h，RIRI组视网膜 Fas⁺细胞表达逐渐增多；再灌注后24 h视网膜Fas⁺细胞数达到高峰，棕色阳性染色细胞分布在视网膜神经节细胞层、内丛状层、内核层及神经纤维层；再灌注后 72 h 视网膜 Fas⁺细胞较再灌注后 24 h 减少。NAS组在再灌注后6 h、12 h、24 h、72 h 视网膜 Fas⁺细胞数均较 RIRI组各时间点减少，再灌注后24 h，Fas⁺细胞数达较高水平，随后下降，差异均有统计学意义（均为P＜0.05）。正常组视网膜可见 FasL 全层低表达。RIRI组再灌注后 6 h，视网膜神经节细胞层和神经纤维层存在少量 FasL⁺细胞；再灌注后12 h FasL蛋白表达逐渐增多；再灌注后24 h FasL⁺细胞数达高峰，可见深棕色的细胞膜及细胞质染色细胞分布在视网膜神经节细胞层、内丛状层、内核层及神经纤维层；再灌注后72 h FasL蛋白的阳性表达逐渐减少。NAS组再灌注后6 h、12 h、24 h、72 h 视网膜FasL⁺细胞数均少于 RIRI组各时间点阳性细胞数，差异均有统计学意义（均为P＜0.05）。结论　NAS可通过抑制RIRI大鼠视网膜细胞Fas、FasL蛋白的表达，减轻缺血再灌注对大鼠视网膜细胞造成的损伤。     

Edaravone inhibits procaspase-3 denitrosylation and activation through FasL-Trx2 pathway in KA-induced seizure

Previous studies have demonstrated that excessive free radicals play an essential role in the initiation and progression of epilepsy and that a novel exogenous free radical scavenger edaravone (Ed) exerts some neuroprotective effects on seizure-induced neuronal damage. The purpose of this study was to elucidate the possible molecular mechanisms of Ed associated with procaspase-3 denitrosylation and activation through the FasL-Trx2 pathway in seizures rats. In this study, we investigated the effects of Ed on the regulation of the combination of Fas ligand/Fas receptor and the major components of the death-inducing signaling complex (DISC) in the hippocampus of kainic acid (KA)-treated Sprague Dawley (SD) rats. Treatment with Ed can attenuate the increased expression of FasL induced by KA and prevent procaspase-3 denitrosylation and activation via suppression of the FasL-Trx2 signaling pathway, which alleviates the neuronal damage in seizures. These results provide experimental evidence that Ed functions by preventing the denitrosylation and activation of procaspase-3 and that Ed acts as a therapeutic option for epilepsy.     

Responsiveness to i.v. immunoglobulin therapy in patients with toxic epidermal necrolysis: A novel pharmaco-immunogenetic concept

Toxic epidermal necrolysis (TEN) represents a rare drug-induced autoimmune reaction with delayed-type hypersensitivity that initiates the process of developing massive keratinocyte apoptosis, dominantly in the dermoepidermal junction. Although the etiopathophysiology has not yet been fully elucidated, the binding of Fas ligand (FasL, CD95L) to the Fas receptor (CD95) was shown to play a key role in the induction of apoptosis in this syndrome. The knowledge of the role of immunoglobulin G (IgG) in inhibition of Fas-mediated apoptosis contributed to the introduction of i.v. Ig (IVIg) in the therapy of TEN patients. Despite great enthusiasm for this therapy at the end of the 1990s, subsequent studies in various populations and meta-analyses could not unequivocally confirm the efficacy of the IVIg-based treatment concept. Today, therefore, we are faced with the dilemmas of how to adjust therapy of TEN patients most effectively, which patients could benefit from IVIg therapy and what dose of the preparation should be administrated. The ground-breaking question is: do the host genetic profiles influence the responsiveness and side-effects of IVIg therapy in TEN patients? Based on recent pharmacological, immunological and genetic findings, we suggest that the variability of IVIg therapy outcomes in TEN patients may be related to functional variants in Fas, FasL and Fc-γ receptor genes. This novel concept could lead to improved quality of care for patients with TEN, facilitating personalized therapy to reduce mortality.     

口腔鳞癌患者手术前后血清可溶性Fas的变化

目的 研究可溶性Fas在口腔鳞癌患者手术前后血清中的浓度变化.方法 选择口腔鳞癌患者25例为鳞癌组,健康志愿者11例为对照组.采用双抗体夹心ELISA法检测口腔鳞癌患者术前和术后第7天以及对照组健康者血清中可溶性Fas的浓度.结果 血清中可溶性Fas的浓度鳞癌组术前为(8.20±1.17) ng/L、术后为(6.45±1.23) ng/L,对照组为(5.62±1.22) ng/L.鳞癌组术前可溶性Fas浓度高于对照组,差异有统计学意义(P＜0.05);鳞癌组手术前后可溶性Fas浓度差异有统计学意义(P＜0.05).结论 血清可溶性Fas水平测定对估计口腔癌的病情变化可能有一定的临床意义.     

Mechanism of Fas-mediated cell death and its enhancement by TNF-alpha in human salivary gland adenocarcinoma cell line HSG

Fas-mediated cell death in a human salivary gland adenocarcinoma cell line (HSG) was induced by treatment of the cells with agonistic anti-Fas antibody (CH-11), and this cell death was enhanced by pretreatment with tumor necrosis factor alpha (TNF-alpha). The mode of cell death was apoptosis, because it was accompanied by caspase activation and the cleavage of poly(ADP-ribose) polymerase. The TNF-alpha treatment of the cells increased the expression of Fas, which was accompanied by the activation of nuclear factor kappaB (NFkappaB). These results suggest that the enhancement of the apoptosis caused by TNF-alpha resulted from increased sensitivity of the HSG cells to CH-11-mediated apoptosis due to induction of Fas protein by TNF-alpha via the activation of NFkappaB. In order to elucidate the apoptosis signaling pathway, we examined the effect of various caspase inhibitors on the apoptosis induced by CH-11. Fas-mediated apoptosis of HSG cells was slightly inhibited by the caspase-9 inhibitor although it was mainly inhibited by that for caspase-8. Based on this finding, we consider CH-11-induced apoptosis in HSG cells to be mainly mediated by the type I death signaling pathway that is caused by a caspase cascade initiated by the activation of caspase-8 at the death-inducing signaling complex (DISC).     

口腔鳞状细胞癌Fas基因表达与细胞凋亡关系的研究

目的　探讨Fas基因mRNA及蛋白表达水平与人口腔鳞状细胞癌细胞凋亡的关系。方法　应用Northern 杂交、双参数流式术(TUNEL法)检测人口腔鳞状细胞癌中Fas mRNA及蛋白的表达,鳞癌细胞的细胞周期与凋亡水平。结果　5例正常口腔黏膜标本中均有Fas mRNA及蛋白的表达。在口腔鳞癌组织中,Fas mRNA及蛋白的表达与口腔鳞癌组织病理学分级有关(P<0·005),与口腔鳞癌组织分化程度和凋亡指数呈正相关。结论　口腔鳞癌中 Fas基因的表达与组织病理学分级及细胞凋亡之间关系密切。     

Fas转染及抗体处理对舌鳞癌细胞Tca8113移植瘤形成和增殖的影响

目的　研究Fas转染和抗Fas单克隆抗体对舌鳞癌细胞系Tca8113裸鼠体内移植瘤形成和增殖的影响,探讨相关机制。方法　脂质体法以真核表达重组质粒pBK-fas转染舌鳞癌细胞系Tca8113。部分转染细胞行抗Fas单克隆抗体处理。未转染细胞、Fas转染细胞、Fas转染抗体处理细胞分别接种裸鼠皮下。观测移植瘤生长,绘制生长曲线。RT-PCR检测肿瘤细胞Fas mRNA表达。流式细胞术(FCM)检测肿瘤细胞凋亡、增殖及Fas蛋白表达。结果　Fas转染和抗体处理延迟肿瘤形成,抑制肿瘤增殖。Fas转染上调移植瘤Fas mRNA表达,提高Fas蛋白表达强度和凋亡指数,但不影响Fas蛋白阳性表达率和增殖指数。抗体处理不影响Fas mRNA和蛋白表达,但可提高凋亡指数,降低增殖指数。结论　Fas转染抑瘤效应是上调Fas转录和表达、促进凋亡的结果。抗Fas单克隆抗体抑瘤效应则与激活凋亡和抑制增殖有关。     

外周血可溶性Fas蛋白及其配体、髓过氧化物酶水平对重症肺炎患儿预后不良的预测价值

目的观察重症肺炎患儿外周血可溶性Fas蛋白（sFas）、可溶性Fas蛋白配体（sFasL）和髓过氧化物酶（MPO）水平变化,并探讨三者对重症肺炎预后不良的预测价值。 方法选取四川中医药高等专科学校绵阳富临医院2016年2月至2020年5月收治的182例重症肺炎、196例轻症肺炎患儿和178例健康儿童,分别为重症组、轻症组和对照组;重症组患儿再根据预后分为预后不良组（29例）和预后良好组（153例）。采用单因素方差分析比较重症组、轻症组治疗前和对照组外周血sFas、sFasL和MPO水平;采用单因素和多因素Logistic回归分析重症组患儿预后不良的影响因素,采用受试者工作特征（ROC）曲线评价外周血sFas、sFasL和MPO水平以及联合指标预测重症组患儿预后不良的价值。 结果三组研究对象的性别、年龄和体重,重症组与轻症组患儿病原微生物分布、肺炎分期差异均无统计学意义（P均> 0.05）。重症组患儿治疗前外周血sFas、sFasL和MPO水平分别为（104.63 ± 19.75）ng/L、（1 062.36 ± 179.85）ng/L和（1 020.26 ± 59.71）U/L,轻症组患儿分别为（80.52 ± 13.66）ng/L、（703.57 ± 127.66）ng/L和（796.75 ± 43.02）U/L,对照组儿童分别为（58.78 ± 10.16）ng/L、（577.83 ± 121.22）ng/L和（632.59 ± 38.71）U/L;重症组和轻症组患儿以上3个指标水平均高于对照组（sFas：重症组 vs.对照组：t = 27.605、P < 0.001;轻症组vs.对照组：t = 17.322、P < 0.001;sFasL：重症组 vs.对照组：t = 29.908、P < 0.001,轻症组vs.对照组：t = 9.744、P < 0.001;MPO：重症组 vs.对照组：t = 71.920、P < 0.001;轻症组vs.对照组：t = 38.647、P < 0.001）,重症组患儿以上3个指标水平均显著高于轻症组（t = 13.885、22.488、41.973,P均< 0.001）。重症组患儿预后不良发生率为15.93%（29/182）。预后不良组患儿双重/多重感染占比（χ² = 12.081、P = 0.001）、多肺叶感染占比（χ² = 32.378、P < 0.001）和外周血白细胞计数（WBC）（t = 6.432、P < 0.001）、中性粒细胞百分比（N%）（t = 3.658、P = 0.001）、C-反应蛋白（CRP）（t = 19.415、P < 0.001）、降钙素原（PCT）（t = 26.101、P < 0.001）、sFas（t = 13.717、P < 0.001）、sFasL（t = 5.357、P < 0.001）和MPO（t = 5.435,P < 0.001）水平均显著高于预后良好组患儿;多因素Logistic回归分析显示以上指标均为重症组患儿预后不良的危险因素,差异均有统计学意义（OR = 5.969、95%CI：4.857~6.304、P = 0.029,OR = 7.485、95%CI：6.785~8.126、P = 0.014,OR = 5.332、95%CI：4.593~5.567、P = 0.010,OR = 4.959、95%CI：4.246~5.337、P = 0.015,OR = 5.143、95%CI：4.879~5.695、P = 0.003,OR = 6.126、95%CI：5.630~6.558、P = 0.008,OR = 8.325、95%CI：6.452~9.902、P = 0.005,OR = 8.469、95%CI：7.879~8.653、P = 0.001,OR = 9.132、95%CI：8.882~9.594,P = 0.003）。外周血sFas、sFasL和MPO水平预测重症组预后不良的Cut-off值分别为125.07 ng/L、1 171.21 ng/L和1 053.04 U/L;sFas、sFasL和MPO以及3个指标联合预测的曲线下面积（AUC）分别为0.875、0.890、0.897和0.955,3个指标联合预测AUC均显著高于sFas、sFasL、MPO水平单独预测,差异均有统计学意义（Z = 5.693、P = 0.005,Z = 5.192、P = 0.007,Z = 4.982、P = 0.009）。 结论重症肺炎患儿外周血sFas、sFasL和MPO水平均偏高,且在预后不良重症患儿中水平均更高,其联合应用可预测患儿不良预后。