皮肤良恶性病变DNA原位定量AgNORs的图象分析

应用彩色病理图象分析仪对６１例皮肤良性恶性病变进行ＤＮＡ原位定量及ＡｇＮＯＲｓ的图象分析，结果表明，皮肤癌的ＤＮＡ倍体多呈出Ｃ５，〉Ｃ５，ＡｇＮＯＲｓ的图象分析示３种变量即ＧＮ（颗粒数目），ＧＡ（颗粒面积）ＧＶ（颗粒的变异系数）意义较大，在良恶性皮肤病中差异非常显著（Ｐ〈０．０１）。研究表明，ＤＮＡ原位定量与ＡｇＮＯＲｓ的图象分析对判断皮肤良恶性病变，皮肤癌的鉴别诊断及癌前病变的预测有重要意义。     

特应性皮炎急、慢性皮肤炎症时IgGFc受体的表达

特应性皮炎 (ＡＤ)是一种以血清中总的和抗原特异性的ＩｇＥ和ＩｇＧ4水平增高为特征的变应性皮肤病 ,在其炎症处有大量Ｔ细胞、树突状细胞 (ＤＣｓ)和巨噬细胞浸润 ,这些细胞在ＡＤ局部炎症反应的调节中发挥着重要作用 ,ＩｇＧ受体 (ＦｃγＲ)参与了ＤＣｓ和巨噬细胞功能。作者以特应性皮肤斑贴试验(ＡＰＴ)阳性处皮肤作为ＡＤ急性炎症模型 ,慢性皮损作为慢性炎症模型对ＡＤ急、慢性变态反应时人皮肤中不同的ＤＣｓ和巨噬细胞亚群ＦｃγＲ细胞的表达进行了体内研究 ,另外由于ＡＰＴ的发生机制可能是特异性或非特异性炎症 ,因此作者对经刺激…     

皮肤鳞癌细胞中Ag－NORs计数与DNA含量的关系

皮肤鳞癌细胞中Ａｇ－ＮＯＲｓ计数与ＤＮＡ含量的关系余春艳，关鹏举，苏勤，谢德豪第四军医大学唐都医院皮肤科（邮政编码７１００３８）本文对１０例皮肤鳞状细胞癌（以下简称鳞癌）病人进行Ｆｅｕｌｇｅｎ细胞光度测定法和核仁组成区相关嗜银蛋白（Ａｇ－ＮＯＲ）计数...     

不同波长紫外线照射后皮肤颜色的变化

目的 观察不同波长紫外线照射皮肤后，颜色变化的过程。方法 用双倍剂量最小持续性黑化量和最小红斑量对10例Ⅲ型受试者皮肤进行照射，通过临床评分、扫描反射比分光光度仪和窄谱反射分光光度计三种方法对照射后的皮肤进行14天的评价和测定。结果 UVB照射后，a*值和红斑指数（EI值）在照射后6 h急剧增加，照射后2天达到高峰；L*值在照射后1天出现急剧降低；ITA°在第7天显著降低；黑素指数（MI值）在照射后2天内有逆向的降低趋势，直到照射后7天才有显著增高。在UVA照射下，a*值和EI值改变不明显；L*值在照射后6 h出现显著降低；ITA°在第14天达到最低值； MI值仅照射后1天有显著增高。结论 UVA和UVB照射后的皮肤颜色改变在时间动力学和反应程度方面有明显区别。a*值和EI值是评价照射后日晒伤较为敏感而准确的参数，而ITA°和MI值是评价晒斑较好的参数。     

银屑病患者皮损表皮生长因子及其受体的检测

用抗表皮生长因子（ＥＧＦ）多克隆抗体、抗表皮生长因子受体（ＥＧＦＲ）单克隆抗体及ＡＢＣ免 疫酶标技术，对２０例寻常型银屑病患者皮损及１０例正常人皮肤进行观察。结果表明：①正常人皮肤及 银屑病非皮损区皮肤ＥＧＦ及ＥＧＦＲ主要分布在表皮基底细胞层及基底层上部。②银屑病进行期皮损 ＥＧＦ及ＥＧＦＲ分布于表皮各层，表皮中、上层含量明显升高。③经有效治疗消退期皮损ＥＧＦ及ＥＧＦＲ 从角质层开始消退，分布趋于正常。提示ＥＧＦ及ＥＧＦＲ对银屑病皮损角肮细胞过度增殖及异常分化起 重要作用。     

原发性皮肤T细胞淋巴瘤的T细胞受体基因重排分析

目的：评价Ｓｏｕｔｈｅｒｎ印迹分析（ＳＢＡ）和聚合酶链反应（ＰＣＲ）检测原发性皮肤Ｔ细胞淋巴瘤（ＰＣＴＣＬ）Ｔ细胞受体（ＴＣＲ）基因重排（ＧＲ）的意义。方法：以ＰＣＲ扩增ＴＣＲγ的结合Ⅴ（可变区）－Ｊ（结合区）序列（ＴＣＲγＰＣＲ）和ＳＢＡ分析ＴＣＲβ链基因（ＴＣＲβＳＢＡ）检测克隆性ＧＲ。结果：蕈样肉芽肿（ＭＦ）：ＴＣＲγＰＣＲ和ＴＣＲβＳＢＡ检测６例ⅡＡ期和７例ⅡＢ期皮损标本的ＧＲ分别为５例和４例以及６例和５例，外周血分别有４例、２例和５例、３例示ＧＲ；而７例ⅠＡ期和１０例ⅠＢ期的ＴＣＲγＧＲ和ＴＣＲβＧＲ皮肤组织为４例、１例和７例、１例，外周血为３例、阴性和４例、１例。１例ＭＦⅡＡ表现为皮病性淋巴结病患者的淋巴结中证实有ＧＲ。疑诊ＭＦ：１１例患者的皮损和外周血标本经ＴＣＲγＰＣＲ检测５例皮肤和３例外周血见ＧＲ。非蕈样肉芽肿、Ｓéｚａｒｙ综合征的ＰＣＴＣＬ：ＰＣＲ和ＳＢＡ显示ＴＣＲＧＲ分别为皮肤组织占９例／１０例和６例／８例，外周血占９例／１０例和６例／１１例。Ｓéｚａｒｙ综合征和淋巴瘤样丘疹病：２例Ｓéｚａｒｙ综合征外周血和其中１例皮肤标本同时见ＴＣＲγＧＲ和ＴＣＲβＧＲ；２例淋巴瘤样丘疹病的皮肤标本     

银屑病皮损c—erbB—1及c—erbB—2原癌基因表达及其意义

用抗ｃ－ｅｒｂＢ－１（ＥＧＦＲ）及ｅｒｂＢ－２（Ｎｅｕ蛋白）原癌基因表达蛋白单抗及免疫组化技术，观察２０例奶悄病上骸１０例正常人皮肤。结果表明：（１）在正常人皮肤表皮中，ＥＧＦＲ基因表达蛋主要分布在基底细胞层，Ｎｅｕ蛋白表达蛋白主要分布在棘层上部及颗粒细胞层，（２）银屑病活动性皮损表地及颗粒层ＥＧＦＲ表达增强，而Ｎｅｕ蛋白表皮两种原癌基因表达异常对皮损形成可能起一定作用。     

银屑病皮损c－erbB－1及c－erbB－2原癌基因表达及其意义

用抗ｃ－ｅｒｂＢ－１（ＥＧＦＲ）及ｃ－ｅｒｂＢ－２（Ｎｅｕ蛋白）原癌基因表达蛋白单抗及免疫组化技术，观察２０例银屑病皮损及１０例正常人皮肤。结果表明：（１）在正常人皮肤表皮中，ＥＧＦＲ基因表达蛋白主要分布在基底细胞层，Ｎｅｕ蛋白表达蛋白主要分布在棘层上部及颗粒细胞层。（２）银屑病活动性皮损表皮棘层及颗粒层ＥＧＦＲ表达增强，而Ｎｅｕ蛋白表达减弱。银屑病皮损表皮两种原癌基因表达异常对皮损形成可能起一定作用。     

皮脂腺及皮下脂肪疾病

２００００４１９ 寻常痤疮患者血清硒值及血浆ＧＳＨ－ＰＸ活力测定／符梅（南通医学院附院皮肤科）／／中国皮肤性病学杂志．－１９９９，１３（３）．－１４６ ４０例，病程１个月～１０年。并以６０例健康成人作为对照。采用原子吸收分光光度计法测定。结果表明，患者组血清硒和ＧＳＨ－ＰＸ均较对照组为低，以脓疱为主的患者其血清硒值较正常对照下降更著。作者结合文献对上述结果进行了讨论。参２（刘彤）２００００４２０ 微生物学检查对寻常痤疮治疗的指导意义／吴剑波（北京医大三院皮肤科）…／／中国皮肤性病学杂志．－１９９９…     

窄波UVB诱导正常皮肤色素斑的研究

目的 观测窄波UVB诱导正常人腹部皮肤色素沉着斑的颜色变化指标。方法 采用窄波UVB治疗仪，以1.5倍的最小红斑量照射30例志愿者的腹部皮肤，形成人工色素沉着斑。在照射前、照射后1，2，4，6，8周，分别用 CM-2600d分光光度计测量局部皮肤的L*、a*、b*、△L、△a、△b、 △E值，以SIAscopy仪检测色素斑处血红蛋白、胶原蛋白、黑素总量，并捕获相应图像。同时，用数码相机拍摄志愿者腹部照片。结果 照射后局部皮肤呈淡灰色，4 h后出现局部潮红，1周后色素沉着最明显，2例受试者照射部位出现细小脱屑，所有受试者均无水疱发生。CM-2600d分光光度计测量照射部皮肤，结果显示，照射后1周，L*下降到最低，a*值升至最高，b*变化不明显。1周后L*值开始升高，而a*值下降，直到照射后8周，各自接近照射前水平。SIAscopy仪检测提示，照射后1周，皮肤血红蛋白增加最明显，以后迅速下降，低于照射前水平。而总黑素含量和胶原蛋白照射后持续缓慢增加，第6周达最高。结论 窄波UVB可诱导正常皮肤色素斑形成，通过观察皮肤色素的变化规律，客观地评估美白产品的功效。     

豚鼠多次皮肤刺激试验结果分析

将２种用途、３种剂型，共３２份化妆品进行多次皮肤刺激试验的结果进行半定量分析，发现多次皮肤刺激指数等分分级同组织病理检查积分等分分级呈直线正相关（ ｒ＝０． ９９７， Ｐ＜０． ０１）。提示可以通过多次皮肤刺激指数分级和组织病理检查积分分级来衡量化妆品刺激性的大小，也可作为化妆品质量优劣的评价标准之一。另外，我们提出不同用途、不同剂型化妆品的刺激性存在明显差异（ Ｐ <０． ０５），故其评价标准应有所区别。     

Value of the cutaneous basophil hypersensitivity (CBH) response for distinguishing weak contact sensitization from irritation reactions in the guinea pig

Numerous studies of the histology of allergic contact dermatitis reactions to potent allergens in guinea pigs and humans have indicated that there is significant tissue infiltration with basophilic leukocytes. In this study we determined whether this histologic finding could be of value in distinguishing weak sensitization reactions from primary irritation, thereby aiding in the predictive identification of weak or moderate contact allergens. Guinea pigs were sensitized by the Buehler test method. Skin reactions were graded 24, 48, and 72 h post-challenge with duplicate patch sites biopsied at the 24- or 72-h grading timepoints. The biopsies were fixed, embedded in glycol methacrylate, thin sectioned, and Giemsa stained. The number of basophils per 400 leukocytes were counted along the upper dermis just below the dermal/epidermal junction. Challenge patch sites from animals sensitized to a relatively low dose of the strong contact allergen, oxazolone, were compared with patch sites from animals challenged only with a strong irritant, sodium lauryl sulfate (SLS). Compared to normal skin (7.5 +/- 1.0 basophils/400 leukocytes +/- SEM) only the oxazolone patch sites showed significant basophil infiltration (36.8 +/- 6.5), despite the fact that the skin reactions to the low oxazolone challenge dose were relatively weak. SLS patch sites showed no basophil infiltration above normal skin levels (4.8 +/- 0.9). Subsequent blinded studies compared weak/moderate presumptive sensitization reactions (as defined by accepted visual skin grading criteria) to various chemicals (citronellal, vanillin, cinnamic aldehyde, and ethylenediamine) to primary irritation reactions to the same chemicals. In each case, low-challenge-dose sensitization sites on previously treated (induced) animals showed mean basophil infiltration (range, 11.9-69.2 basophils/400 leukocytes) significantly greater than higher-dose irritant reactions (range, 1.6-13.3). The range for normal skin was 0.2-10.2 and the range for strong patch reactions to higher concentrations of oxazolone was 59.8-209.3. These data strongly indicate that light-microscopic quantitation of the CBH response can be used to distinguish relatively weak to moderate contact sensitization reactions from primary irritation reactions to the same chemicals.     

Evaluation of modifications of the traditional patch test in assessing the chemical irritation potential of feminine hygiene products

Background/aims: The first main objective of the work presented in this paper was to investigate ways of optimizing the current arm patch test protocol by (1) increasing the sensitivity of the test in order to evaluate more effectively the products that are inherently non‐irritating, and/or (2) reducing the costs of these types of studies by shortening the protocol. The second main objective was to use the results of these studies and the results of the parallel studies conducted using the behind‐the‐knee method to better understand the contribution of mechanical irritation to the skin effects produced by these types of products. In addition, we were interested in continuing the evaluation of sensory effects and their relationship to objective measures of irritation. Methods: Test materials were prepared from three, currently marketed feminine protection pads. Wet and dry samples were applied to the upper arm using the standard 24‐h patch test. Applications were repeated daily for 4 consecutive days. The test sites were scored for irritation prior to the first patch application, and 30–60 min after removal of each patch. Some test sites were treated by tape stripping the skin prior to the initial patch application. In addition, in one experiment, panelists were asked to keep a daily diary describing any sensory skin effects they noticed at each test site. Results: All protocol variations ([intact skin/dry samples], [compromised skin/dry samples], [intact skin/wet samples], and [compromised skin/wet samples]) gave similar results for the products tested. When compared to the behind‐the‐knee test method, the standard upper arm patch test gave consistently lower levels of irritation when the test sites were scored shortly after patch removal, even though the sample application was longer (24 vs. 6 h) in the standard patch test. The higher level of irritation in the behind‐the‐knee method was likely due to mechanical irritation. The sensory skin effects did not appear to be related to a particular test product or a particular protocol variation. However, the mean irritation scores at those sites where a sensory effect was reported were higher than the mean irritation scores at those sites were no sensory effects were reported. Conclusions:

• ? All four protocol variations of the standard upper arm patch test can be used to assess the inherent chemical irritant properties of feminine protection products.
• ? For these products, which are inherently non‐irritating, tape stripping and/or applying wet samples does not increase the sensitivity of the patch test method.
• ? Differences in irritation potential were apparent after one to three 24‐h applications. Therefore, the standard patch test protocol can be shortened to three applications without compromising our ability to detect differences in the chemical irritation produced by the test materials.
• ? The patch test can be used to evaluate effectively the inherent chemical irritation potential of these types of products. However, this method is not suitable for testing the mechanical irritation due to friction that occurs during product use.
• ? There is no relationship between specific test conditions, i.e., compromised skin and/or testing wet samples and reports of perceived sensory reactions.
• ? However, there seems to be a clear relationship between sensory reactions and objective irritation scores.

     

Comparison of objective and sensory skin irritations of several cosmetic preservatives

There are many cosmetic ingredients, such as preservatives and fragrances, known to elicit adverse effects. The aim of this study was to investigate the side-effects of cosmetic preservatives, by evaluating objective and subjective skin irritation. The method comprised of 2 parts. In part 1, we tried to compare 24-hr patch test results with the sensory irritation potential of several preservatives. In part 2, skin cumulative irritation test for 21 days and sensory irritation test were performed to compare various combinations of preservatives in 4 types of formulations. Our data showed that methylparaben, ethylparaben, propylparaben, butylparaben, phenoxyethanol (PE) and chlorphenesin (CPN) have similar objective skin irritation potential at the minimal inhibitory concentration of each preservative, but CPN has higher potential than other preservatives in subjective irritation. Sensory irritation of preservatives changed according to formulation type, and PE combined with CPN highly increased irritation. There was correlation between antimicrobial activity and skin objective irritation but not sensory irritation. Influence on skin sensory irritation varies with the combination of preservatives. Therefore, for the development of new preservatives and cosmetics, it is important to evaluate skin sensory irritation of preservatives used in cosmetic products according to the type of formulations.     

A two-center study of the development of acute irritation responses to fatty acids.

BACKGROUND: A human 4-hour patch test has been developed for evaluating the acute irritation potential of chemicals. This method was developed for comparative irritation assessments. Although skin irritation responses in human subjects can be quite variable, this test method has proven robust in both intra- and interlaboratory tests. However, the previous interlaboratory studies were not optimal in that slightly differing protocols were used and the studies were not controlled for time of year or source of test chemicals. As a result, some variation in acute irritation responses were seen that might have been reduced somewhat had these variables been controlled to a greater extent. OBJECTIVE: The purpose of the current study was to examine interlaboratory reproducibility of the 4-hour patch test when conducted under as identical as set of test conditions as possible. METHODS: Two laboratories conducted a direct comparison study of the acute irritation potential of three structurally related, undiluted fatty acids (octanoic acid, decanoic acid, and dodecanoic acid) in comparison to a benchmark positive control chemical (20% sodium dodecyl sulfate [SDS]). The studies were run within a 4-month period using the same commercial source of test chemicals. Test subjects were treated with each chemical under occluded patch conditions for gradually increasing exposure duration up to 4 hours. The results were then evaluated in terms of total cumulative incidence of positive responses and time response patterns. RESULTS: Using statistical comparisons of the proportion of the subjects with a positive irritant reaction to each substance, the rank order of irritation potential was decanoic acid >/= octanoic acid > SDS > and dodecanoic acid. The statistical comparisons and the time-response patterns for each chemical were nearly identical at the two laboratories. There were also very similar, and intriguing, variations in the interchemical response patterns seen in the two studies. CONCLUSION: When conducted under as controlled a set of test conditions as was reasonably possible, this acute irritation protocol shows remarkably high consistency across independent test laboratories, further supporting its continued development and acceptance as a valid and more predictive tool for assessing skin irritation potential.     

Quantitative evaluation of patch test reactions: a comparison between visual grading and erythema index image analysis

Background/aims: The interpretation of patch test reactions may vary between examiners. As test results are graded, an issue also arises when differing degrees of erythema are placed in the same grade. The purpose of this study was to quantitatively evaluate the degree of erythema in patch tests using image analysis and to study the usefulness of this method by comparing it with visual grading. Methods: A total of 121 Japanese patients were patch tested with various materials. At 48 h, digital photographs of the patch test areas were taken, in addition to a visual evaluation by dermatologists. Digital images of the areas were converted to erythema index (EI) images using image processing and both EI and ΔEI (the difference between the patch test site and adjacent normal skin) values of the patch test sites were compared with the corresponding visual grades. Results: An excellent linear correlation (r=0.95) was found between ΔEI and visual grades, although EI also significantly correlated with visual grades. There were significant differences (P<0.0001–0.05) between the mean ΔEI values of any two adjacent visual grades. Conclusion: ΔEI values derived from image processing appear to be suitable for the quantitative evaluation of erythema in patch tests. This method may be helpful in overcoming the subjectiveness of visual evaluation and for training non‐experts in patch testing.     

Topical isotretinoin in Darier's disease

Topical 0·05% isotretinoin (Isotrex ® gel) was used to treat a test patch of skin in 11 patients with Darier's disease. Hyperkeratosis and papules improved in six patients after treatment for 3 months. Erythema, burning and irritation were common adverse effects, and these were severe in three patients, one of whom stopped treatment. Patients with mild Darier's disease may find topical isotretinoin helpful, but it is likely that most patients with widespread disease will require treatment with systemic retinoids.     

Evaluation of skin susceptibility to irritancy by routine patch testing with sodium lauryl sulfate

Irritant patch testing with sodium lauryl sulfate (SLS) will become more and more a routine test determining skin susceptibility in men. Recently, it has been shown that for practical reasons, irritant SLS patch testing can take place on the back simultaneously with a routine allergic patch test to other contact allergens. However, SLS patch testing has mostly been performed on the forearm in studying experimental skin irritation so far. The aim of this study was to determine whether there is a relationship in skin response to aqueous SLS (0.125%; 0.25%; 0.5% and 1.0%) between the forearm and the back assessed by visual scoring and measurement of transepidermal water loss (TEWL). We found a pronounced reaction of the forearm compared to the back. TEWL values as well as visual scores correlated well with SLS concentration. There was also a high correlation in visual scoring between the forearm and the back. Based on test sensitivity and specificity we suggest a 48 hrs patch test for routine screening with 0.5% SLS on the forearm evaluated by TEWL measurement or visual scoring 24 hrs after patch removal. A mild erythema (scored as < or =1) is considered to be normal. If for practical reasons, the SLS patch is placed on the back simultaneously with the allergic patch test, 0.5% SLS may be sufficient, too. TEWL measurement so far provides a reliable method and will certainly be necessary for experimental studies on irritant skin reactions, particularly when different SLS concentrations are used. After a 48 hrs patch test with SLS 0.5% TEWL measurement should be performed at 72 hrs. A value of < or =31.6 g/m(2)hr seems to follow the normal distribution.     

Evaluation of human skin irritation by carboxylic acids, alcohols, esters and aldehydes, with nitrocellulose-replica method and closed patch testing

Closed patch testing and the nitrocellulose replica method are performed as useful clinical methods for the evaluation of human skin irritation by cosmetics and topical medicaments. Comparison of the sensitivity between microscopic scoring by nitrocellulose-replica method and visual scoring by closed patch test in the detection of skin irritation, however, has not been well studied with statistical analysis. Here, we evaluated human skin irritation by carboxylic acids. Alcohols, esters and aldehydes, with different chain length (C8-C18), using both methods. The results of closed patch testing showed that, although the score of skin irritation for carboxylic acids (C8, C12), alcohols (C8) and aldehydes (C8), tested at a concentration of 0.5 m -2.0 m . significantly increased with increasing concentration of the test compounds, ester compounds scarcely caused any irritation on the surface of the skin occluded. In addition, an increase of carbon chain length in the test compounds made it impossible to detect skin irritation. In contrast, the nitrocellulose-replica method could evaluate skin reactions against very weak irritants that gave no macroscopic alterations on the skin surface in the closed patch test. However, the scoring system is somewhat subjective and should be improved to make the analysis more objective.     

Patch tests versus use tests in skin irritation risk assessment

In the risk assessment of the ability of a substance or preparation to cause skin irritation, a patch test involving human volunteers is often seen as providing definitive information. However, the skin exposure conditions found in an occluded patch may be far removed from those in a particular use situation, not least for such reasons as the high dose per unit area, the duration of exposure and the maceration of the skin. Whilst these factors may enhance sensitivity in the identification of intrinsic hazard, they may do little to ensure accurate safety evaluation for real use. In this paper, we report data from a series of studies with an unmarketed facial skin cosmetic product. Whilst the product was unexpectedly highly irritating in a standard patch test in 30 volunteers, subsequent use tests, including a 6× daily open application to the elbow for 3 weeks and 2× daily application in a half-face test lasting 3 to 4 weeks and involving 52 volunteers failed to show any evidence of skin irritation. It is concluded that the most meaningful results for skin irritation risk assessment are likely to come from studies which involve relevant patterns of exposure.