| 1. The clinical analysis of port wine-stains (PWS) treated by hematoporphyrin monomethyl ether-photodynamic therapy (HMME-PDT) Objective:To evaluate the efficacy and side effects of PWS treated by HMME-PDT. Methods:Fifty five patients entered the study.5 mg/kg HMME was slowly and continuously intravenous injected for 20 minutes,10 minutes after the HMME started to inject, the PWS lesion was irradiated by continuous KTP 532 nm laser for 20 minutes (80-100mW/cm2,7cm spot size). Results:The response rate was 74.5%. The excellent response rate of the pink lesion was higher than that of the purple lesion(x2=12.432,P＜0.001). The response rate of the female was higher than that of the male (x2=9.319,P=0.002). The excellent response rate of the lesion which received 2 PDT-treatments was higher than that of the lesion which received only 1 treatment. (x2=7.922, P=0.005). The degree of crusting after the second treatment was less than that after the first treatment (P＜0.001).21 patients (38.2%) had hyperpigmentation and one patient (1.8%) had hypopigmentation. no long-term side effects were found. Conclusions:The HMME-PDT on PWS was efficacy and safety with a higher clearance rate of lesions.2. The relationship of lesion-response and efficacy of HMME-PDT on PWS Background:HMME-PDT was successfully used on PWS treatment, but always companied with some seriously pain and crusting several days later. Objective:To study the possibility of the relationship between the pain response, crusting and the efficacy of HMME-PDT on PWS. Methods:Twenty six PWS patients were involved in the study and received 1 treatment.5mg/kg HMME was slowly and continuously intravenous injected, then the PWS lesion was irradiated by continuous KTP 532 nm laser for 20 min. The patients were asked to report when the pain emerged and how serious it was. The severity of crusting was also observed after treatment. Two months after treatment, the efficacy was judged based on the pre-and post-photographs. The relationship between efficacy and pain-response, crusting was analyzed. Results:PWS treated by HMME-PDT always companied with severe pain, and the degree of pain aggravated gradually when the treatment was going on(r=0.848, P＜0.001), and the pain alleviated immediately when the irradiation stopped. But we failed to found any relationship between the degree of pain, crusting and the efficacy. Conclusion:Neither the extent of pain nor the crusting could influence the efficacy of HMME-PDT on PWS.3. The factors influencing HMME-PDT on the vascular endothelial cell (EC-304) in vitro.Objective:To observe the possible relationship between the death of EC-304 after HMME-PDT and the HMME-incubating-time, the HMME-incubating-concentration, and the laser energy density. Methods:The EC-304 was incubated with different incubating-time and different incubating-concentration of HMME, and then irradiated by continuous KTP 532 nm Laser with different energy density. After PDT, The cell was judged by trypan blue and then counted under microscope. Results:After PDT, no effect on the EC-304 was found after only 1 min incubating time with HMME comparing with the control group (P>0.05). Elongating the HMME-incubating time (5-30 minutes) would aggravating the EC-304 death by PDT (P＜0.05), but the PDT effect of all the groups on EC-304 was almost the same (P>0.05). Comparing with the control group, HMME-PDT resulted in significant EC-304 death (P＜0.05), but on the same laser fluency (energy density), the effect was almost the same in different groups even with different irradiating time (120-300 seconds) or the different irradiance (25.5-63.7mW/cm2). In some range, the higher of the HMME dosage it was, the fewer of the EC-304 survived (r=-0.890, P＜0.01). Conclusions:All of the HMME-incubating-time, HMME concentration and the laser energy density were very important for HMME-PDT on EC-304.4. The effect of PDT on TNF-αsecreting by EC-304Objective:To observe the effect of HMME-PDT on TNF-αsecreting. Methods:EC-304 was incubated in 6-well plates. Four groups were designed: HMME-PDT group, HMME control group, laser control group, blank control group. The supernatant of experimental groups were harvested at 12 hours,24 hours, 48 hours after PDT respectively. The supernatant of the blank control group was also harvested at the same time after exchanging the culture solution. The output of TNF-αwere measured with ELISA. Results:In the different harvesting point, the TNF-αoutput of the groups were different. In generally, the output of the 3 control groups were the similar (P=0.72), but the HMME-PDT group shown significant higher output of TNF-αcomparing with any of the other control groups (P＜0.05). Conclusions:HMME-PDT can promote the EC-304 to secrete TNF-a.
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