ECP-treated lymphocytes of chronic graft-versus-host disease patients undergo apoptosis which involves both the Fas/FasL system and the Bcl-2 protein family

Chronic graft-versus-host disease (cGVHD) is a severe and frequent complication of allogenic bone marrow transplantation which is often treated with extracorporeal photochemotherapy (ECP) with a positive clinical outcome in patients resistant to conventional protocols. The mechanism of action of ECP has not been fully elucidated, although several authors have reported that it is able to induce apoptosis. Using samples obtained from ten cGVHD patients, we sought to determine whether lymphocytes treated with ECP underwent apoptosis and, above all, the mechanisms involved. Lymphocytes at four stages were isolated: immediately before ECP, from the last buffy coat collected, after UV irradiation prior to reinfusion, and the day after ECP. When cultured for 48 h, lymphocytes treated with ECP underwent accelerated apoptosis (tested as annexin V binding cells and as intracellular histone-associated DNA fragments) in comparison with lymphocytes from the other samples. This enhanced programmed cell death could not be prevented by IL-2. Immediately after isolation, there was no difference in Bcl-2 or bax expression among the four different samples, or in Fas and FasL mRNA. However, when cultured, lymphocytes treated with ECP showed a rapid downregulation of Bcl-2, an upregulation of bax with an increased bax/Bcl-2 ratio, a decrease in bcl-2 mRNA and an increase in Fas. No changes were detectable in lymphocytes from the other samples. IL-2 and TNF- production was not significantly different among lymphocytes from the four samples. In conclusion, in patients affected by cGVHD, ECP induced apoptosis of lymphocytes with the involvement of both the Fas/FasL system and the Bcl-2 protein family.F.L.P. and M.F. contributed equally to this work.     

Treatment of polymorphous light eruption

Summary This study was designed in order to prove on a large-scale basis the efficacy of oral photochemotherapy (PUVA) in the prevention of polymorphous light eruption (PLE), to work out indication criteria for PUVA treatment of this disease, and to establish a simple method based on anamnestic data to differentiate UVA from UVB induced PLE. The results obtained in 106 PLE patients (85 UVA-, 21 UVB-induced) demonstrate that: 1) time consuming phototesting for determination of the disease's action spectrum is unnecessary for practical purposes; 2) PUVA-induced tanning under routine conditions represents a potent prophylaxis even in severe cases of PLE; and 3) topical sunscreens (sunblockers) in the majority of cases are sufficient to protect UVB-promoted PLE, but fail in UVA-induced disease. PUVA thus seems the treatment of choice only in UVA-mediated PLE, the action spectrum of the disease at least in most cases being easily discovered from certain anamnestic data.     

5-氨基酮戊酸光动力疗法治疗寻常痤疮的临床观察

目的观察5-氨基酮戊酸光动力疗法（ALA-PDT）治疗寻常痤疮的安全性及有效性。方法将90例中重度痤疮患者随机单盲分为两组：治疗组45例,给予ALA-PDT治疗,1次/10 d,共治疗2～4次;对照组45例,口服异维A酸胶囊,每次10 mg,2次/d,共治疗40 d。在治疗的第10、20、30及40天对两组患者进行疗效判断和比较,同时,记录治疗过程中出现的不良反应。结果治疗组患者经过2～4次治疗后,总有效率为97.8%;对照组治疗结束时总有效率为77.8%,治疗组疗效明显优于对照组（P〈0.01）。另外,治疗组复发程度明显低于对照组,且病情控制时间明显延长。治疗组的所有皮损,包括粉刺、丘疹、脓疱、囊肿、结节等均比对照组明显减少。结论 ALA-PDT治疗中重度痤疮的疗效明显优于口服异维A酸胶囊,是一种简单、高效、不良反应轻微的治疗中重度寻常痤疮的新方法。     

Benzochloroporphyrin Derivative Induced Cytotoxicity and Inhibition of Tumor Recurrence During Photodynamic Therapy for Osteosarcoma

Photodynamic therapy (PDT) is a promising cancer treatment modality that uses dye-sensitized photooxidationof biologic matter in target tissue. This study explored effects of the photosensitizer BCPD-17 during PDT forosteosarcoma. LM-8 osteosarcoma cells were treated with BCPD-17 and cell viability after laser irradiationwas assessed in vitro with the 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide assay. The effectsof BCPD-17 during PDT recurrence were then examined on tumor-bearing mice in vivo. BCPD-17 had dosedependentcytotoxic effects on LM-8 osteosarcoma cells after laser irradiation which also had energy-dependenteffects on the cells. The rate of local recurrence was reduced when marginal resection of mice tumors was followedby BCPD-17-mediated PDT. Our results indicated BCPD-17-mediated PDT in combination with marginalresection of tumors is a potentially new effective treatment for osteosarcoma.     

5-氨基酮戊酸光动力疗法预防尖锐湿疣复发的临床研究

目的对尖锐湿疣治疗后局部检测人乳头瘤病毒（HPV）阳性患者行光动力治疗,观察其清除病毒、预防复发的作用。方法尖锐湿疣患者经CO2激光或冷冻治疗祛除疣体,并在创面愈合后,在原皮损及周围2cm处行HPV检测阳性者60例,随机分两组,治疗组采用ALA-PDT治疗,每周1次,连续3次,随后每1～2周随访观察,连续3个月;对照组不用任何药物,每1～2周随访观察,连续3月。ALA-PDT疗程结束后治疗组及对照组患者于原病变部位取材,再次行荧光PCR检测HPV DNA。观察两组治疗区域病毒清除及疣体复发情况。结果治疗组病毒清除率80%,对照组病毒清除率30%,治疗组和对照组之间清除率差异有统计学意义（χ2=15.152,P=0.000）。治疗组在原疣体部位及照射区域复发4例,而对照组17例复发,复发率分别为13.33%和56.67%,治疗组和对照组复发率之间差异有统计学意义（χ2=12.381,P=0.000）。结论 ALA-PDT对尖锐湿疣的复发和控制HPV的潜伏感染都十分重要。     

半剂量PDT治疗45岁以上中心性浆液性脉络膜视网膜病变疗效观察

目的：观察半剂量维替泊芬光动力疗法(photodynamic therapy,PDT)治疗45岁以上中心性浆液性脉络膜视网膜病变(central serous chorioretinophathy,CSC)的有效性。

方法：收集2012-01/2016-12在我院就诊的45岁以上CSC患者30例32眼,平均年龄54.34±8.58岁; 病程1wk～48mo,中位数6mo。所有患眼均采用国际标准视力表行最佳矫正矫正视力(best corrected visual acuity,BCVA)检查,并转换为最小分辨角对数(LogMAR)视力; 采用光学相干断层扫描(optical coherence tomography,OCT)测量黄斑中心凹视网膜厚度(central macular thickness,CMT)及记录视网膜下液(subretinal fluids absorption,SRF)吸收情况; 采用荧光素钠血管造影(fluorescence fundus angiography,FFA)观察患眼脉络膜血管渗漏形态。所有患眼均行半剂量维替泊芬PDT,治疗后1、3、6mo随访观察BCVA和黄斑区OCT情况。对比分析治疗前后患眼BCVA、CMT及SRF吸收变化情况。

结果：所有患者经PDT治疗后1、3、6mo治愈率分别为44%、63%、75%,差异具有统计学意义(χ²=6.621,P=0.037)。将所有患者按病程分为两组,A组患者病程<6mo,B组病程≥6mo,两组患者治疗前BCVA分别为0.6149±0.4117、0.8167±0.4370; 治疗前CMT分别为409.47±129.422、395.82±153.756μm。经治疗后两组间BCVA值差异无统计学意义(F=0.303,P=0.823),但具有时间差异性(F=32.837,P<0.001)。A组患者的BCVA治疗后1mo与治疗前比较,差异具有统计学意义(t=3.347,P=0.005); 治疗后1、3mo比较,差异有统计学意义(t=4.768,P<0.001); 治疗后3、6mo相比,差异无统计学意义(t=2.024,P=0.063)。B组患者的BCVA治疗后1、3、6mo与治疗前两两比较,差异均具有统计学意义(P<0.05)。治疗后两组间患者CMT差异无统计学意义(F=0.064,P=0.978),但具有时间差异性(F=26.447,P<0.001),A组患者的CMT治疗后1、3、6mo与治疗前两两比较,差异均具有统计学意义(P<0.05); B组患者的CMT治疗后1、3、6mo与治疗前两两比较,差异均具有统计学意义(P<0.05)。

结论：半剂量PDT疗法可有效改善45岁以上CSC患者的视力,对视网膜下液的吸收仍有较好疗效。     

T-cell responses in photoimmune therapy

The extracorporeal inactivation of a lymphocyte rich buffy coat suspension with ultraviolet A light and 8 methoxypsoralen can lead to dramatic clinical improvements following reinfusion of the damaged cells. This therapy is reviewed in the context of the disease it is most commonly used for: cutaneous lymphoma. Studies with cutaneous lymphoma patients have shown an active immune response against purified tumor cells. In addition a mouse model for an impact of therapy on a T-cell lymphoma has demonstrated results that parallel those from clinical studies in humans. The impact of photoimmune therapy on in vivo and in vitro T-cell responses to cutaneous lymphoma is discussed.     

Treatment of cutaneous T-cell lymphoma with extracorporeal photochemotherapy

Background Cutaneous T-cell lymphoma (CTCL) and its leukemic erythrodermic form (Sézary syndrome) are malignancies cies of CD4+ T lymphocytes. Extracorporeal photochemotherapy (ECP) selectively affects autoreactive as well as malignant T lymphocytes. The efficacy of ECP depends strongly upon adequate serum/buffy coat levels of the photosensitizer 8-methoxypsoralen (8-MOP). The resorption of orally applied 8-MOP vanes inter- and intraindividually within a broad range, meaning that adequate therapeutic drug levels cannot always he achieved. Therefore, since July 1994 we have exclusively used a liquid 8-MOP preparation which is added directly into the buffy coat fraction of the ECP circuit, resulting in constant high drug levels of approximately 190 ng/ml. Twelve CTCL-patients (six with Sézary syndrome, six with mycosis fungoides received between six and 25 ECP treatments. Some of them had undergone previous therapy without success. Results All patients with Sézary syndrome declared a distinct reduction in intensity of pruritus. Three patients who received liquid 8-MOP extracorporeally showed a partial remission on the basis of skin scores. Of the patients receiving 8-MOP orally, two remained clinically unchanged and one showed a progression of the disease. In these cases, subtherapeutic 8-MOP plasma levels were often found. Of the six mycosis fungoides patients one achieved complete and two partial remission: another two patients showed minor response. These five patients were treated with liquid 8-MOP. One patient showed no change in skin lesions; he had received 8-MOP orally and achieved subtherapeutic photosensitizer plasma levels. Conclusion Our treatment protocols confirm the beneficial effects of ECP on CTCL at any stage, but it seems that adequate ECP efficiency is ensured only when an 8-MOP solution is applied extracorporeally.