Anti-PD1 antibody and not anti-LAG-3 antibody improves the antitumor effect of photodynamic therapy for treating metastatic breast cancer

Photodynamic therapy(PDT)has limited effects in treating metastatic breast cancer.Immune checkpoints can deplete the function of immune cells;however,the expression of immune checkpoints after PDT is unclear.This study investigates whether the limited e±cacy of PDT is due to upregulated immune checkpoints and tries to combine the PDT and immune checkpoint inhibitor to observe the e±cacy.A metastatic breast cancer model was treated by PDT mediated by hematoporphyrin derivatives(HpD-PDT).The anti-tumor effect of HpD-PDT was observed,as well as CD4þT,CD8þT and calreticulin(CRT)by immunohistochemistry and immunofluorescence.Immune checkpoints on T cells were analyzed byflow cytometry after HpD-PDT.When combining PDT with immune checkpoint inhibitors,the antitumor effect and immune effect were assessed.For HpD-PDT at 100 mW/cm2 and 40,60 and 80 J/cm2,primary tumors were suppressed and CD4þT,CD8þT and CRT were elevated;however,distant tumors couldn't be inhibited and survival could not be prolonged.Immune checkpoints on T cells,especially PD1 and LAG-3 after HpD-PDT,were upregulated,which may explain the reason for the limited HpD-PDT effect.After PDT combined with anti-PD1 antibody,but not with anti-LAG-3 antibody,both the primary and distant tumors were signi-cantly inhibited and the survival time was prolonged,additionally,CD4þT,CD8þT,IFN-þCD4þT and TNF-þCD4þT cells were signi-cantly increased compared with HpD-PDT.HpD-PDT could not combat metastatic breast cancer.PD1 and LAG-3 were upregulated after HpD-PDT.Anti-PD1 antibody,but not anti-LAG-3 antibody,could augment the antitumor effect of HpD-PDT for treating metastatic breast cancer.

Efficacy of 1/3-Dose Verteporfin Photodynamic Therapy for Subacute Central Serous Chorioretinopathy

Purpose: To study the safety and efficacy of 1/3-dose verteporfin photodynamic therapy (PDT) for subacute central serous chorioretinopathy (CSC). Methods: In this case series, 59 eyes (59 patients) diagnosed with subacute CSC in Shenyang the 4th hospital from January 2014 to December 2015 were treated with 1/3-dose verteporfin PDT and followed up for at least 1 year. The symptoms and the diagnosed history were more than 3 months but shorter than 6 months. The central foveal thickness (CFT), neuroretinal thickness (NRT), height of subfoveal retinal fluid (SRF), and subfoveal choroidal thickness (SCT) were observed at baseline and after treated at 1, 2, 3, 6 and 12 months with EDI-OCT, Best-corrected visual acuity ( BCVA) was also studied at the same time. Results: After 1, 2, 3 and 6 months of 1/3-dose verteporfin PDT treatment, the BCVA improved significantly (P 0.05). The height of SRF changed significantly. There was no retinal pigment epithelium atrophy and choroidal neovascularization (CNV) in all cases after more than 12 months follow-up. Conclusion: Treatment of 1/3 dose verteporfin PDT could safely and effectively reduce expansion of choroidal vessel and choroidal choriocapillary, promoting absorbance of subretinal fluid for subacute CSC. 1/3-dose verteporfin PDT may be an alternative method to treat the subacute CSC.

应用1/3剂量光动力学疗法治疗急性CSC

目的:观察1/3剂量光动力学疗法(photodynamic therapy,PDT)治疗急性中心性浆液性脉络膜视网膜病变(centralserous chorioretinopathy,CSC)的短期疗效。方法:选取急性CSC患者26例26眼,行单次1/3量维替泊芬(2mg/m2)进行PDT治疗,术前和术后1,4,12wk进行最佳矫正视力和OCT的检测,且于术前、术后4wk和12wk进行眼底荧光素血管造影(FFA),吲哚青绿血管造影(ICG)检查,观察治疗的有效性和安全性。结果:术后1wk,26眼中有20眼(77%)视网膜下积液吸收,其余6眼(23%)视网膜下液部分吸收;术后4wk,26眼视网膜下积液全部吸收,22眼荧光渗漏完全消失,脉络膜血管高渗透性消失;术后12wk,26眼均病情平稳,无复发。治疗后1wk,最佳矫正视力从术前平均0.41升高至0.80。随访期间26眼均未见任何不良反应。结论:1/3剂量PDT治疗急性CSC短期内安全有效。

血卟啉对人胰腺癌细胞Panc-1的光动力作用及机制

目的:研究光敏剂血卟啉的光动力作用对人胰腺癌细胞Panc-1的体外杀伤效应及其主要机制。方法:将光敏剂浓度和光照剂量2个因素按不同水平分组,以CCK-8实验的吸光度值为检测指标并转换为细胞存活率,研究这2个因素对光动力作用的影响及其规律。在此基础上,先后以荧光显微镜、流式细胞仪、Westernblotting测定这一过程中细胞内活性氧的水平、细胞坏死及凋亡比率以及caspase-3蛋白的表达,探讨光动力作用的主要机制。结果:随着光敏剂浓度和光照剂量的增加,光动力治疗(PDT)后Panc-1细胞存活率相应下降,并分别在光敏剂浓度10 mg/L、光照剂量20 J/cm2达到最大杀伤效应。光敏剂浓度和光照剂量之间对降低细胞存活率有协同作用。单独给予光敏剂和光照均不对细胞存活率产生影响。PDT作用可使Panc-1细胞内产生活性氧,并随光敏剂浓度和光照剂量的增加而增多。PDT后细胞出现凋亡和坏死,二者比例随光敏剂浓度和光照剂量的增加而增加,但始终表现为凋亡率>坏死率。PDT使细胞内caspase-3表达增强,并随光敏剂浓度和光照剂量的增加而增加。结论:血卟啉光动力治疗对人胰腺癌细胞株Panc-1具有明确的杀伤作用,但是光敏剂和激光照射本身并不具有独立的杀伤效应。光敏剂浓度和光照剂量2个影响因素在一定范围内与PDT效应之间呈正相关的关系,二者之间存在协同作用。PDT的作用机制主要在于其产生的各种活性氧成分,通过各种途径激活caspase-3,最终导致细胞凋亡。

A型肉毒毒素联合光动力疗法在增生性瘢痕中的应用效果分析

目的 观察A型肉毒毒素(BTXA)联合光动力疗法(PDT)治疗增生性瘢痕(HS)的临床效果方法 选取2018年5月至2020年5月南阳市第二人民医院收治的70例HS患者作为研究对象,按照不同治疗方法将其分为研究组(35例)与对照组(35例),研究组患者采用BTXA联合PDT治疗,对照组患者单纯采用BTXA治疗,对比两组患者温哥华瘢痕量表(VSS)评分、磷酸化信号转导与转录激活因子3 (p-STAT3)及早期生长反应因子-1 (Egr-1)表达水平、临床疗效以及不良反应发生情况结果 治疗4个月,研究组患者VSS评分及p-STAT3、Egr-1表达水平均明显低于对照组(t=4.665、10.583、11.549,P<0.001);研究组患者中显效19例、有效11例、无效5例,明显优于对照组患者的显效12例、有效10例、无效13例(Z=-2.129,P=0.033);研究组患者不良反应发生率为20.00%,与对照组患者的不良反应发生率11.43%无明显差异(χ^(2)=0.971,P=0.324)。结论 BTXA联合PDT治疗HS,可降低瘢痕组织中p-STAT3、Egr-1的表达水平,有效促进瘢痕组织消退,改善瘢痕症状,提高治疗效果,且安全性较高,值得临床推广应用。

低剂量5-氨基酮戊酸光动力疗法对HaCaT细胞皮肤屏障相关蛋白表达的影响

目的通过研究低剂量5-氨基酮戊酸(5-aminolevulinic acid,ALA)光动力疗法(photodynamic therapy,PDT)对HaCaT细胞中皮肤屏障相关蛋白表达的影响,探讨低剂量光动力改善皮肤屏障功能的潜在机制。方法以0、0.01、0.05、0.1、0.5及1 mmol/L ALA孵育HaCaT细胞后分别予以0、0.5、1.85、3.72及10 J/cm^(2)的(635±3)nm红光照射,CCK-8检测细胞存活率,摸索低剂量ALA-PDT促进HaCaT细胞活力的最佳ALA浓度及照光能量参数。将细胞分为对照组、光照组、ALA组、低剂量PDT组,分别予以相应处理后,用实时荧光定量PCR、Western blot实验检测细胞中丝聚合蛋白(filaggrin,FLG)、内皮蛋白(involucrin,IVL)、水通道蛋白3(aquaporin 3,AQP3)、紧密连接蛋白-1(claudin-1,CLDN1)、闭锁蛋白(occludin,OCLN)mRNA及蛋白表达。结果不同ALA浓度及不同照光能量处理HaCaT细胞的结果显示,在0.1 mmol/L ALA及3.72 J/cm^(2)照光能量的条件下,细胞的活力最佳。以该低剂量ALA-PDT处理HaCaT细胞后,FLG、IVL、AQP3、CLDN1及OCLN mRNA的相对表达量分别为10.25±6.34、5.10±2.74、4.89±1.33、8.53±6.43、4.06±2.01,对照组为1.01±0.01、1.02±0.02、1.01±0.01、1.01±0.00、1.02±0.01,光照组为2.64±1.04、2.29±1.49、2.49±1.87、4.51±4.41、1.57±1.12,ALA组为3.03±2.35、1.78±0.90、1.55±1.13、3.72±5.88、1.39±0.89,其中低剂量PDT组中FLG及AQP3表达明显升高,差异有统计学意义(P<0.05)。Wes-tern blot显示低剂量PDT组HaCaT细胞中FLG、IVL、AQP3、CLDN1蛋白表达水平均明显高于对照组、光照组及ALA组(P<0.05),OCLN的表达在几组中差异无统计学意义(P>0.05)。结论0.1 mmol/L ALA联合3.72 J/cm^(2)的照光能量为低剂量ALA-PDT对HaCaT细胞的最适ALA浓度及照光能量;低剂量ALA-PDT可能通过提高部分皮肤屏障相关蛋白的表达起到改善皮肤屏障的作用。