本文選題：鮮紅斑痣 + 光動力��； 參考：《北京協(xié)和醫(yī)學院》2011年博士論文

【摘要】：第一部分光動力、脈沖染料激光及強脈沖光治療鮮紅斑痣的臨床療效觀察 目的評價和比較光動力(PDT)、脈沖染料激光(PDL)和強脈沖光(IPL)治療鮮紅斑痣的臨床療效。方法PDT以血卟啉單甲醚為光敏劑,532nm連續(xù)激光為光源,PDL為595nm激光,IPL根據(jù)不同皮損選擇560nm,590nm或640nm的濾光片,回顧性總結(jié)和分析不同性別、皮損類型及皮損部位與三種治療的療效關(guān)系。結(jié)果患者總數(shù)共130例,其中PDT患者35例,PDL患者56例,IPL患者39例。三種治療組在性別、皮損類型及治療部位的構(gòu)成比上無統(tǒng)計學差異(p分別為0.904、0.929及0.987)。總體療效PDT最高,其次為PDL,最后為IPL,三者的顯效率分別為為54.3%、33.9%和20.5%；不同性別之間三種治療的療效無顯著差異(p分別為0.225、0.821和0.145)；PDT組粉紅型療效明顯優(yōu)于紫紅型(p=0.021)和增厚型(p=0.026),紫紅型和增厚型之間療效無差異(p=0.068)；PDL組及IPL組三種皮損類型之間療效均無差異(PDL組p分別為0.226、0.400和0.946；IPL組p分別為0.803、0.095和0.069)；PDT和PDL組中頸部療效優(yōu)于面部(p分別為0.002和0.004),IPL組的療效無差異(p=0.097)。結(jié)論PDT是治療鮮紅斑痣安全有效的方法,總體療效優(yōu)于PDL和IPL。 第二部分血管內(nèi)皮細胞和角質(zhì)形成細胞對血卟啉單甲醚吸收特性的研究 目的觀察和比較血管內(nèi)皮細胞(ECV304)和角質(zhì)形成細胞(HaCaT)對光敏劑血卟啉單甲醚(HMME)的吸收特性。方法取對數(shù)生長期的ECV304和HaCaT分別與50、100、150、200、250μg/ml的HMME孵育,孵育時間為16h；將150μg/ml的HMME分別與上述兩種細胞孵育,孵育時間分別為15min、30min、lh、3h、8h、12h、24h。激光掃描共聚焦顯微鏡檢測熒光強度。結(jié)果濃度依賴性結(jié)果顯示ECV304的平均熒光強度分別為74.00、125.57、135.24、141.99、132.09；HaCaT的平均熒光強度分別為93.88、102.45、112.59、108.23、10,1.70。時間依賴性結(jié)果顯示ECV304的平均熒光強度分別為95.07、103.97、105.96、108.99、112.93、115.36、122.91；HaCaT的平均熒光強度分別為104.25、106.60、108.72、113.75、117.66、114.90、118.14。結(jié)論ECV304和HaCaT對HMME的吸收在一定濃度范圍和時間范圍內(nèi)均呈孵育濃度和孵育時間依賴性。 第三部分血卟啉單甲醚在血管內(nèi)皮細胞和角質(zhì)形成細胞中的定位及光動力治療靶點的研究 目的探討血卟啉單甲醚(HMME)在血管內(nèi)皮細胞系ECV304和角質(zhì)形成細胞系HaCaT的亞細胞定位及光動力治療的作用靶點。方法HMME與ECV304和HaCaT分別孵育1h和18h,熒光探針分別標記線粒體、核膜、細胞膜和過氧化物酶體,觀察不同時間點下HMME與細胞器的結(jié)合率；HMME與ECV304和HaCaT分別孵育20h,給予532nm連續(xù)激光照射,比較照光前后上述細胞器中HMME的熒光強度變化。結(jié)果HMME與細胞器的結(jié)合率實驗結(jié)果顯示,ECV3041h組線粒體、核膜、細胞膜、過氧化物酶體HMME的結(jié)合率分別為1.18、0.72、0.95、0.68,18h組分別為1.35、0.83、0.73、0.91; HaCaTlh組分別為1.09、0.66、0.92、0.77,18h組分別為1.13、0.86、0.72、1.10。HMME在細胞器中的漂白率實驗結(jié)果顯示,ECV304分別為18.22%、10.77%、7.44%、8.56%, HaCaT分別為11.90%、5.02%、3.82%、8.90%。結(jié)論ECV304 HMME主要定位于線粒體,HaCaT定位于的線粒體和過氧化物酶體；ECV304光動力的主要作用靶點可能為線粒體,而HaCaT的主要作用靶點可能為線粒體和過氧化物酶體。
[Abstract]:The first part is photodynamic therapy, pulsed dye laser and intense pulsed light in the treatment of port wine stains.
Objective to evaluate and compare the clinical efficacy of photodynamic (PDT), pulsed dye laser (PDL) and strong pulse light (IPL) in the treatment of nevus fresh red spot. Methods PDT was used as a photosensitizer with hematoporphyrin monomethyl ether, 532nm continuous laser as light source, PDL as 595nm laser, IPL based on different skin lesions to select 560nm, 590nm or 640nm filters, and retrospective summary and analysis of different sex, The relationship between the type of skin lesion and the site of skin lesion and the curative effect of three kinds of treatment. The total number of patients was 130 cases, including 35 cases of PDT, 56 cases of PDL and 39 cases of IPL. There was no statistical difference between the three treatment groups in the sex, type of skin lesion and the position of treatment (P respectively 0.904,0.929 and 0.987). The overall effect was the highest, followed by PDL, and finally I. PL, the effective rates of the three were 54.3%, 33.9% and 20.5%, and there was no significant difference between the three treatments (P 0.225,0.821 and 0.145 respectively). The efficacy of the PDT group was obviously superior to the purple red type (p=0.021) and the thickening type (p=0.026), and there was no difference between the purple and the thickening types (p=0.068), and three types of skin lesions in PDL and IPL groups. There was no difference in the effect between the types (group PDL, P, 0.226,0.400 and 0.946, P in group IPL, 0.803,0.095 and 0.069), and in group PDT and PDL, the effect of the neck was better than that of the face (P 0.002 and 0.004 respectively), and there was no difference in the efficacy of IPL group (p=0.097). Conclusion PDT is an effective method for the treatment of nevus of fresh erythema.
The second part is about the absorption characteristics of vascular endothelial cells and keratinocytes to hematoporphyrin monomethyl ether.
Objective To observe and compare the absorption characteristics of vascular endothelial cells (ECV304) and keratinocyte (HaCaT) on the photosensitizer, hematoporphyrin monomethyl ether (HMME). Methods the logarithmic growth period ECV304 and HaCaT were incubated with HMME of 50100150200250 mu g/ml respectively, and the incubation time was 16h, and the HMME of the 150 micron /ml was incubated with the above two cells respectively. The fluorescence intensity was detected by 15min, 30min, LH, 3h, 8h, 12h, 24h. laser scanning confocal microscopy. The result of concentration dependence showed that the average fluorescence intensity of ECV304 was 74.00125.57135.24141.99132.09, and the average fluorescence intensity of HaCaT was 93.88102.45112.59108.23,10,1.70. time dependent results showing ECV304. The average fluorescence intensity is 95.07103.97105.96108.99112.93115.36122.91, and the average fluorescence intensity of HaCaT is 104.25106.60108.72113.75117.66114.90118.14. conclusion ECV304 and HaCaT HMME absorption in a certain concentration range and time range are both incubation concentration and incubation time dependence.
The third part is the localization of hematoporphyrin monomethyl ether in vascular endothelial cells and keratinocytes and the target of photodynamic therapy.
Objective to investigate the subcellular localization and photodynamic target of hematoporphyrin monomethyl ether (HMME) in vascular endothelial cell line ECV304 and keratinocyte line HaCaT. Methods HMME and ECV304 and HaCaT were incubated with 1H and 18h respectively. The fluorescent probes labeled mitochondria, nuclear membrane, cell membrane and peroxisome respectively, and observed HMME at different time points. The binding rate of organelles; HMME and ECV304 and HaCaT were incubated for 20h respectively. 532nm continuous laser irradiation was given, and the fluorescence intensity of HMME in the above-mentioned organelles before and after illumination was compared. Results the binding rate of HMME and organelles showed that the binding rate of mitochondria, nuclear membrane, cell membrane and peroxisome HMME in ECV3041h group was 1.18,0.72,0., respectively 1.18,0.72,0.. The 95,0.68,18h group was 1.35,0.83,0.73,0.91, and the results of the bleaching rate of 1.13,0.86,0.72,1.10.HMME in the group 1.09,0.66,0.92,0.77,18h group respectively showed that the ECV304 was 18.22%, 10.77%, 7.44%, 8.56%, and HaCaT were 11.90%, 5.02%, 3.82% respectively, and 8.90%. concluded that ECV304 HMME was mainly located in the mitochondria, HaCaT determination. Mitochondria and peroxisomes are located; the main target of ECV304 photodynamic may be mitochondria, and the main target of HaCaT may be mitochondria and peroxisomes.
【學位授予單位】：北京協(xié)和醫(yī)學院
【學位級別】：博士
【學位授予年份】：2011
【分類號】：R758.51

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