美白祛斑药物对强脉冲光作用下的鼠皮黑色素的影响

观察美白祛斑药物对强脉冲光(intensive pulse light, IPL)作用下的鼠皮黑色素的影响,探讨IPL作用下,皮肤的黑色素的增生和美白药物之间的作用关系,以期探寻美白药物在减少光子嫩肤术并发症的作用机制.以小白鼠作为研究对象,用IPL在一定的波长及能量密度照射活体小鼠皮肤, 在照射后15 min涂抹低浓度的维A酸霜,分别在照射后的4 h、5 d,用显微镜观察涂霜剂、没涂霜剂的皮肤黑色素及其周边组织的变化情况;在照射后的1 d用多光子显微镜观察涂霜剂、没涂霜剂皮肤组织的变化.讨论了美白祛斑药物在光子嫩肤过程中抑制黑色素生成的过程,为如何减少光子嫩肤术并发症提供参考和借鉴.     

基于OCT技术的鼠皮肤激光热损伤修复过程的监测

采用Er:YAG激光(波长为2 940 nm,能量密度为:2.5 J/cm2单光斑,扫描次数为4)照射活体小白鼠皮肤,利用光学相干层析成像(optical coherence tomography,OCT)技术在活体小鼠上观察其皮肤组织在激光作用之前及作用之后光热损伤修复的整个过程,得到了激光光热作用下引起损伤的皮肤组织在此过程中皮肤光学特性参数的变化情况,发现皮肤修复过程中光学参数有显著差异,并分析了这些差异引起的原因,以揭示激光美容中并发症主要因素。     

血卟啉单甲醚对鸡冠皮肤光动力学效应的研究

目的　本文采用组织形态学观察方法 ,比较血卟啉单甲醚 (ＨＭＭＥ)与ＨｐＤ对鸡冠表皮层、真皮浅层毛细血管网和真皮深层组织的光敏损伤特点及影响因素。材料与方法　成年莱亨鸡 2 58只 ,静脉注射ＨＭＭＥ或ＨｐＤ 1～ 2 0ｍｇ ｋｇ ,于给药后即刻～ 4小时对鸡冠进行激光照射 ,波长为 4 88.0～ 514.5ｎｍ、510 .6ｎｍ、578.2ｎｍ或 6 2 7.8ｎｍ ,功率密度为 50～ 150ｍＷ ｃｍ2 ,能量密度为 30～ 2 70Ｊ ｃｍ2 ,于照射后即刻、1、3、7、14和 2 8天取材 ,进行大体、光镜和透射电镜观察。结果　ＰＤＴ作用区的红色鸡冠完全变白 ,乳头层毛细血…     

国产Nd:YAP激光组织热损伤效应的比较研究

目的:通过比较1 341 nm Nd:YAP与临床常用激光器脉冲Nd:YAG和连续Nd:YAG对皮肤、肌肉、静脉和肝脏组织的热损伤效应,了解Nd:YAP激光的作用特点,为其临床应用提供实验依据。方法:以大鼠作为实验对象,分为皮肤、肌肉组和静脉、肝脏组。皮肤、肌肉组:Nd:YAP激光、连续和脉冲Nd:YAG选择能量密度1000 J/cm2、2 000 J/cm2,静脉、肝脏组采用Nd:YAP激光和连续Nd:YAG两种激光器,能量密度选择500 J/cm2和1 000 J/cm2,对组织进行非接触式点状照射。照射过程中观察组织变化和损伤直径大小,并于照射后即刻、3天取材做病理切片,光镜下观察病理变化,比较相同能量密度下三种激光器对组织的热损伤和损伤修复情况。结果:皮肤、肌肉组:Nd:YAP激光在1 000 J/cm2条件下对组织的损伤以热凝固效应为主,2 000 J/cm2条件下组织发生明显气化、有一定的切割作用。相同能量密度下,脉冲Nd:YAP激光气化作用较两种Nd:YAG激光显著,而凝固损伤范围较连续Nd:YAG浅,较脉冲Nd:YAG深。肝脏、静脉组:Nd:YAP激光对肝脏的热凝固效应与Nd:YAG激光近似,气化作用明显;Nd:YAP激光对血管及其周围组织的凝固及气化作用均较连续Nd:YAG激光明显。结论:Nd:YAP激光具有良好的凝固与气化作用。     

蓝光对浮游状态和生物膜内铜绿假单胞菌的抗菌作用

目的：探讨450 nm-470 nm可见光（蓝光）是否具有杀灭浮游状态和生物膜内铜绿假单胞菌的作用。方法：分别采用不同能量密度的蓝光照射浮游状态铜绿假单胞菌,与红光对照组、空白对照组相比,将照射后细菌采用平板涂板法评价蓝光杀菌效果;制作铜绿假单胞菌生物膜模型,16 J/cm2能量密度蓝光照射后通过激光共聚焦显微镜和扫描电子显微镜观察生物膜内细菌存活情况以及生物膜结构变化。结果：与空白对照组相比,2 J/cm2及以上能量密度组蓝光照射后,细菌数目明显减少,杀菌率明显增加（P〈0.05）,并呈剂量效应关系;16 J/cm2能量密度光照后生物膜内细菌死亡数较空白对照组明显增加且生物膜结构变稀疏。结论：450 nm-470 nm可见光（蓝光）具有高效杀灭浮游状态和生物膜内铜绿假单胞菌的作用。     

SD大鼠急性放射性皮炎模型的建立

目的:建立安全的SD大鼠急性放射性皮炎模型。方法:将28只雄性SD大鼠随机分为7组,分别为空白对照组,电子线照射组(60 Gy,45 Gy,30 Gy),X线照射组(45 Gy,30 Gy,15 Gy),每组4只。选择臀背部皮肤,去毛后照射。放疗后第一天起开始观察动物皮肤表现,采用Douglas and Fowler评分方法记录每只动物皮炎情况,并定期测量动物体重,观察动物一般情况并记录死亡情况。于放疗后第28天处死动物,取照射区域皮肤行HE染色及免疫组化染色(CD3,CD11c,CD68,IV型胶原),以通过光镜分析射线照射后皮肤组织变化情况、真皮层内炎症细胞浸润类型及胶原形成情况。结果:至放疗后第28天X线照射组动物出现大量死亡,电子线照射组动物均存活,电子线各组均出现不同程度的放射性皮炎反应,镜下可见局部组织不同程度的表皮层坏死、炎症细胞浸润、毛囊及附属器减少等表现,以电子线照射60 Gy及45 Gy组表现明显。免疫组化结果显示放射线照射可使真皮层内以CD68为表面标志的巨噬细胞浸润增加,并促进以IV型胶原为标志胶原细胞形成。结论:电子线60 Gy及45 Gy照射SD大鼠臀背部皮肤可建立一种安全有效的急性放射性皮炎动物模型,其临床表现及病理表现可用于实验研究。     

Q开关激光爆破技术对豚鼠皮肤色素的影响

目的研究Q开关激光爆破术对豚鼠黑素细胞的影响及照射周边组织的变化,为临床治疗皮肤色素病变提供实验依据。方法用Q开关-YAG激光分别照射豚鼠黑色毛区及棕色毛区(波长分别用1064nm和530nm,光斑直径2mm),实验动物20只,随机分四组,分别于照射后间隔7d、10d、14d取材,照射前取材作对照,10%甲醛固定,冰冻切片,分别用HE和DOPA反应显示黑素细胞。结果照射后皮肤黑色素颗粒逐渐减少至消失,照射后30d黑毛区与棕毛区肉眼见照射区皮肤变白,毛也变白,HE染色皮肤、毛囊及毛未见黑色素颗粒,DOPA反应表皮黑色素细胞、毛囊和毛均呈阴性反应,部分豚鼠棕毛区毛及毛囊见黄色色素颗粒。结论波长1064nm和532nm Q-YAG激光对豚鼠皮肤黑色素细胞和黑色素颗粒的破坏效果显著;但对棕色色素清除效果较差。波长1064nm Q-YAG激光对豚鼠皮肤黑色素消减与照射次数有关,与照射间隔时间长短无关。     

小鼠皮肤及其毛囊早期发育的组织学观察

目的探讨小鼠皮肤及其毛囊的早期发育规律。方法采用常规石蜡切片和H-E染色技术,观察昆明系小鼠出生前后皮肤及其毛囊的形态发育。结果(1)孕龄16 d胎鼠的皮肤表面形成凹凸不平的深褶皱,但在生后3 d~5 d不仅皱褶的数量减少,而且凹陷变浅;(2)胎鼠孕龄16 d至19 d,其皮肤的表皮、真皮及皮肤总厚度呈现平稳增厚。但是,出生后,其表皮、真皮和皮肤总厚度急剧降低;在生后第1天至第9天,表皮呈现平稳增厚,而真皮则在生后快速厚度,第7天达到最高值(1861.50μm);(3)孕龄16 d的胎鼠皮肤中可观察到初级毛囊,至生后第7天其密度呈现平稳增长;与其相比,次级毛囊从18 d胎鼠开始出现,其密度增长非常迅速,出生后第7天达到1257.14/mm;毛囊的总密度与次级毛囊呈现相似的变化趋势。出生第7天后,由于毛囊的数量急剧增加,无法观察初级毛囊和次级毛囊的变化规律;(4)初级毛囊和次级毛囊的长度与深度变化在出生前后的相对缓慢,与其相比在第3天以后至第7天呈现迅速变化趋势。结论小鼠皮肤及其毛囊的生长性发育发生在胎儿晚期和生后的早期,而其周期性变化可能从出生后的第9天以后开始出现;在孕期16 d至生后第7天可能是检测毛囊特异性基因表达的最佳期。     

微等离子束对豚鼠皮肤超微结构分析和羟脯氨酸测定

目的观察微等离子束对豚鼠皮肤胶原组织作用效应的组织学和超微结构变化及羟脯氨酸含量测定,探讨微等离子束的作用机理。方法选择15只豚鼠,每只豚鼠背部划分为实验侧和空白对照侧2个等分区域,给予60W/10 kJ微等离子束照射,于作用后即刻、1周后和1月后分别切取实验侧及空白对照部位皮肤行组织病理维多利亚-立春红染色,透射电镜分析和羟脯氨酸检测试剂盒进行含量测定。结果 60 W/10 kJ即刻表现为表皮局灶性出现点阵状改变,部分表皮出现汽化缺失或者坏死变性,真皮浅层胶原组织出现点阵化表现和明显均质化;特殊染色显示微等离子束主要影响真皮胶原纤维,形成局灶性胶原纤维凝集和变性。1周后皮肤浅层胶原组织结构逐渐致密,排列有序,有少量组织细胞。1月后皮肤浅层胶原组织明显增厚,胶原纤维增粗并排列致密,弹力纤维呈局灶性增粗。透射电镜显示微等离子束作用后表皮细胞较完整,细胞间结构正常,但真皮胶原丧失正常结构,细胞结构消失,大量细胞凋亡明显,1月后仍可见少量细胞凋亡的表现但胶原结构逐渐恢复,浅层胶原纤维排列明显致密。羟脯氨酸测定显示微等离子束作用1周后羟脯氨酸含量要高于作用之前,但是差异性不具有统计学意义（P〉0.05）;1月后羟脯氨酸含量要明显高于作用前,差异性具有统计学意义（P〈0.05）。结论微等离子束对豚鼠皮肤胶原组织作用有明显的刺激效应,其主要靶组织为真皮胶原组织,可以明显促进皮肤新生胶原的增生。     

不同能量激光照射后中华大蟾蜍早期胚胎发育及其蛋白质谱变化的研究

应用YAG倍频激光器选择不同能量密度激光微束对中华大蟾蜍4细胞期胚胎进行照射,观察胚胎的生长、发育状况,统计出膜率、畸胎率和死亡率。实验结果表明：低能量激光照射明显地提高了胚胎的孵化速度,且使畸变率降低;而高能量激光对胚胎发育有明显抑制作用,而且使畸变率增高。不同能量密度激光微束照射中华大蟾蜍早囊胚,0．5h后制备样品进行双向电泳,凝胶成像后用PDQuest软件进行分析。结果显示激光照射对蛋白质点数及其分布有影响,影响程度与照射激光密度有关。结论：激光照射可以影响胚胎发育和蛋白质的合成。     

The revascularization of pedicle skin flaps in pigs: a functional and morphologic study

Functional and morphologic changes occurring during the revascularization of pedicle flaps have been investigated in the skin of pigs. The skin flaps, 16 cm long by 4 cm wide, were based on a row of segmental vessels arising from the internal mammary artery. Comparative measurements were made in flapped and normal skin. The inherent blood supply in the pedicle of the flap was unable to maintain the whole of the flap in a viable state. Flap viability was ascertained at surgery by the use of the intravital dye Disulphine blue. Injections of the dye after surgery gave a less accurate prediction of viability than when dye was injected prior to surgery. Revascularization between the flap and surrounding skin was evident 3 to 4 days postoperatively at the distal, most hypoxic part of the viable flap. The whole flap had a collateral vascular supply 7 to 10 days after surgery. Isotope clearance studies showed that the greatest functional changes occurred in the distal third of the viable flap, where, after initially slowing, the clearance rate became faster than in normal skin (day 5). Potassium extraction studies indicated similar changes. However, an increase in the red-cell volume on day 1 suggested that vascular shunting was occurring. The results of the morphologic studies indicated a correlation between the number of blood vessels per unit area, the thickness of the dermis, and the recorded functional changes. Seven days after surgery, when isotope clearance rates were very rapid, there was a significant increase in the vascular density and dermal thickness.     

Wound healing in X-irradiated mammalian skin.

Certain parameters of wound healing were investigated in mouse skin given 950 rads and 3325 rads of X-irradiation at various times relative to wounding. Increased inflammation, delayed dermal regeneration, and delayed contraction were noted in all irradiated groups. The activation of surrounding hair follicles, a process that usually accompanies wounding in mouse skin, occurred earlier, over a shorter elapsed time, and over a greater area of skin in animals irradiated prior to wounding than in the controls or in those irradiated and wounded simultaneously. Epidermal mitotic activity in wounds made at the time of irradiation was initially depressed but recovered by the second postoperative day. Wounds in pre-irradiated animals gave an unexpected result. They responded with an immediate burst of mitotic activity without the usual 24-hr lag that was seen in controls. In the pre-irradiated specimens a substantial number of cells appeared to die after dividing.     

Effect of fibroblast implants on wound healing of irradiated skin: assay of wound strength and quantitative immunohistology of collagen

The role of dermal fibroblasts in the expression of radiation-induced damage to the skin was studied. Fibroblasts from neonatal mice were cultured, harvested, and injected into full-depth surgical incisions in the dorsal area of mouse skin, which had been previously locally irradiated by 18 Gy X rays. As a control, cells irradiated with a dose of 20 Gy were also injected. The effect of radiation and fibroblast implants on the gain of skin wound strength was assayed. In an additional experiment freshly isolated cells were implanted. Two weeks following wounding the irradiated skin had reached only about a third of the strength of unirradiated skin. A significant increase of wound strength in irradiated skin was observed when 1.5-2 x 10(6) cultured fibroblasts or freshly isolated fibroblasts were injected into the 20-mm-long wound bed. Irradiated cells had significantly less effect. This suggests that implanting isolated syngeneic cells may "rescue" wounds from the effect of prior irradiation. Semiquantitative immunohistology of types I and III collagen was performed in parallel using a video image digitizing system. Levels of both types I and III collagen were altered in the dermis and the wound tissues in irradiated skin, but the implant of cultured fibroblasts did not affect notably the total levels and the disposition of the two collagen isotypes.     

Chromosomal abnormalities and spleenocyte production in laboratory mouse males after exposure to stress chemosignals

Activity of antibody producing spleenocytes and chromosome stability in bone marrow cells from laboratory mouse males of CBA strain after exposure to different chemosignals excreted by stressed or irradiated syngeneic donors was studied. It has been shown that the exposure of the recipient males to volatiles from donor males (stressed by swimming) decreases quantity of antibody-producing cells in 1, 3 and 10 days after the treatment. The same exposure increased the chromosome aberrations level in dividing bone marrow cells from CBA recipients in 1 day after the treatment. Similar changes were observed in 24 h after exposure to volatiles of irradiated donors or to synthetic mouse pheromone, 2,5-dimethylpyrazine. Possible mechanisms of chemosignals effect on the immune system are discussed.     

Effects of gamma-radiation on the differentiation of mouse melanocytes in the hair follicles

Pregnant mice were whole-body irradiated with a single acute dose of gamma-rays (60Co) to investigate the effect of gamma-radiation on embryonic melanoblasts. The effect was studied by scoring changes in the differentiation of melanocytes in the hair follicles of mice heterozygous for the recessive coat color mutation pink-eyed dilution (p). Abnormal round melanocytes were found in the hair matrix and the dermal papilla of F1 offspring 3.5 days after birth. However, these round melanocytes possessed a melanin deposition of similar intensity to normal hair follicular melanocytes. The frequency of the abnormal hair follicles increased in a dose-dependent manner. Moreover, higher frequencies were found in the animals irradiated at earlier stages of embryonic development. These results indicate that gamma-radiation affects dendritogenesis and the location of mouse melanocytes in the hair follicles, with greater effects seen at the earlier stages of development.     

Reparative regeneration of rat skin under influence of hollow cathode lamp (HCL) with manganese and copper line spectrum emission

The influence of local light exposure by a hollow cathode lamp with a typical manganese and copper (HCL-Mn, Cu) line emission spectrum on the posttraumatic regeneration of rat skin has been investigated. We performed a comparative analysis of the morphology and differentiation potential of rat skin on the 15th and 24th days after a full-thickness skin wound was made to the dorsum. The injured area was irradiated for 30 s every day for 2 weeks. Fifteen days after the loss of the scab, reepithelialization and the recovery of hair growth were monitored (visual observations), unlike in the control rats, which still had their scabs at 24 days. Histological analysis revealed that, as differs from the nonirradiated control group, upon exposure to HCL-Mn, Cu resulted in an increased number of hair follicles and sebaceous glands, as well as a decreased number of blood vessels and a horizontal orientation of collagen fibers. Immunohistochemical assay with antibodies to dendritic cell marker OX-62 revealed the highest cell number of dermal dendritic cells 15 days after exposure; at 24 days, their number was decreased. In the control group, the number of dermal dendritic cells was significantly lower. Immunohistochemical assay with pan-keratin antibodies 15 days after the surgery revealed a high number of cells that express different types of keratins distributed in most parts of the epidermis in control animals, whereas, in the experimental group, this number was significantly lower and concentrated closer to the external part of the epidermis. The number of keratin 19-positive cells in experimental animals was higher 15, rather than 24, days after the surgery as compared to control rats. Thus, manganese and copper spectrum emissions stimulate innate immunity; accelerate the recovery of the derma, skin epithelium, and other skin derivates; and facilitates wound healing.     

Safety and toxicological evaluation of a novel niacin-bound chromium (III) complex

Chromium is an essential trace element required for normal protein, fat and carbohydrate metabolism. It also helps in energy production and increasing lean body mass. Niacin-bound chromium (NBC) is a unique form of bioavailable chromium that promotes healthy lipid profile. This study was focused on determining the broad spectrum safety of NBC. Acute oral, acute dermal, primary dermal irritation and primary eye irritation toxicities of NBC were evaluated. Ames bacterial reverse mutation assay, mouse lymphoma test and a dose-dependent 90-day subchronic toxicity were also conducted. In safety studies, the acute oral LD(50) of NBC was found to be greater then 5000 mg/kg in both male and female Sprague-Dawley rats. No changes in body weight or adverse effects were observed following necropsy. The acute dermal LD(50) of NBC was found to be >2000 mg/kg. The primary skin irritation test was conducted with NBC on New Zealand Albino rabbits. NBC was classified as slightly irritating. The primary eye irritation test was conducted with NBC on rabbits. NBC was classified as practically non-irritating to the eye. NBC did not induce mutagenic effects in the bacterial reverse mutation test in five Salmonella typhimurium strains (TA1535, TA98, TA100, TA97a and TA102), either with or without metabolic activation. Similarly, NBC did not induce mutagenic effects in the mammalian cell gene mutation test in L5178Y mouse lymphoma cells TK (+/-), either with or without metabolic activation. A dose-dependent 90-day subchronic toxicity study demonstrated no significant changes in selected organ weights individually and as percentages of body and brain weights. NBC supplementation did not cause changes in hepatic lipid peroxidation or DNA fragmentation after 30, 60 or 90 days of treatment. Hematology, clinical chemistry and histopathological evaluations did not show any adverse effects in all organs tested. Taken together, the above results indicate a broad spectrum of safety for NBC.     

A comparative analysis of deferoxamine treatment modalities for dermal radiation-induced fibrosis

The iron chelator, deferoxamine (DFO), has been shown to potentially improve dermal radiation-induced fibrosis (RIF) in mice through increased angiogenesis and reduced oxidative damage. This preclinical study evaluated the efficacy of two DFO administration modalities, transdermal delivery and direct injection, as well as temporal treatment strategies in relation to radiation therapy to address collateral soft tissue fibrosis. The dorsum of CD-1 nude mice received 30 Gy radiation, and DFO (3 mg) was administered daily via patch or injection. Treatment regimens were prophylactic, during acute recovery, post-recovery, or continuously throughout the experiment (n = 5 per condition). Measures included ROS-detection, histology, biomechanics and vascularity changes. Compared with irradiated control skin, DFO treatment decreased oxidative damage, dermal thickness and collagen content, and increased skin elasticity and vascularity. Metrics of improvement in irradiated skin were most pronounced with continuous transdermal delivery of DFO. In summary, DFO administration reduces dermal fibrosis induced by radiation. Although both treatment modalities were efficacious, the transdermal delivery showed greater effect than injection for each temporal treatment strategy. Interestingly, the continuous patch group was more similar to normal skin than to irradiated control skin by most measures, highlighting a promising approach to address detrimental collateral soft tissue injury following radiation therapy.