非节段型白癜风患者皮损单细胞转录图谱的初步研究

【摘要】 目的 采用单细胞RNA测序技术鉴定和区分白癜风皮损真表皮细胞亚群，并研究它们之间的关系。方法 2019年9月在杭州市第三人民医院皮肤科门诊收集2例健康人（无免疫及系统性疾病）正常皮肤和2例非节段型稳定期白癜风患者皮损样本，采用10 × Genomics单细胞RNA-Seq技术检测，对所有样本的11 000个细胞进行单细胞转录组测序。通过Seurat软件分析、筛选和统计细胞亚群。结果 对2例正常皮肤基因表达的聚类分析发现包括角质形成细胞、成纤维细胞、神经及黑素细胞、内皮细胞、组织干细胞和以树突细胞及T细胞为主的免疫细胞群。2例白癜风皮损中分化和数量异常的有成纤维细胞和4类角质形成细胞亚群，其中，成纤维细胞占比为0，低于正常皮肤（0.4%），角质形成细胞亚群5、6、10、12占比（8.03%、7.36%、3.52%、0.91%）均显著高于正常皮肤（4.47%、3.53%、2.69%、0.28%，均P＜0.01）。上述亚群的角质形成细胞处于细胞分化的末端，同时还具有极显著且特异的标记基因，分析显示，标记基因主要与细胞间相互作用和细胞稳态密切有关，GO和KEGG分析显示，角质形成细胞亚群5、6主要与细胞间连接和细胞黏附及细胞骨架功能相关，角质形成细胞亚群10与细胞稳态紧密相关。结论 国内首次报道通过单细胞测序手段研究白癜风皮损的转录表达谱，初步发现4群数量及功能差异的角质形成细胞，提示角质形成细胞亚群的异常分化与功能异常可能影响白癜风的发生发展。     

Th17细胞在非节段型白癜风发病机制中的作用

自身免疫调节异常是非节段型白癜风发病的主要机制,Th17细胞作为新发现的CD4+T辅助性T细胞（Th细胞）亚群,可打破自身免疫耐受,导致自身免疫性黑素细胞破坏,在白癜风发病中起重要作用。本文就Th17细胞在白癜风发病机制中所起作用加以综述。     

毫火针联合308nm准分子光治疗面部稳定期非节段型白癜风的疗效观察

目的:观察毫火针联合308 nm准分子光治疗面部稳定期非节段型白癜风的疗效及安全性。方法:68例面部稳定期非节段型白癜风患者共68处白斑(面部白斑最长处直径为3～6 cm),每一处白斑沿最长直径平均分成两半,一半设为治疗侧,先进行毫火针治疗,10 min后,整个白斑同时照射308 nm准分子光;未行毫火针治疗的另一半设为对照侧。308 nm准分子光每周照射2次,毫火针每2周1次,连续治疗观察12周,治疗结束2周后评价疗效。结果:治疗侧与对照侧总有效率分别为97.06%(66/68)、79.41%(54/68),差异有统计学意义(x~2=10.20,P0.05)。治疗侧与对照侧治疗次数(以308nm准分子光治疗次数计算)分别为(7.21±1.25)次、(10.71±2.11)次,差异有统计学意义(t=11.77,P0.05)。所有患者在治疗过程中均未出现严重不良反应。结论:毫火针联合308 nm准分子光治疗面部稳定期非节段型白癜风有效而且安全,值得临床推广。     

节段型白癜风患者同侧出现白发1例

患者女,32岁.因右面部色素脱失斑3年于2008年11月于我科就诊.患者于2005年1月右面部出现色素脱失斑,未引起患者重视,缓慢逐渐增多,并出现右眼睫毛以及右侧胡须变白.患者在出现色素脱失斑前皮肤无瘙痒,也无外伤.     

节段型白癜风特殊性、辨治体会及验案举隅

白癜风是一种常见难治性皮肤顽疾，诸多白癜风患者体质辨识证候难循，而节段型白癜风发病部位、年龄、诱因等皆有其特殊性，中医证候、辨证治疗较非节段型更具特点。本文探讨节段型白癜风的特殊性及中医辨治体会，并报告1例典型病例，强调四诊合参、体察入微的重要性。     

白癜风125例临床分析及与HLA-DRB1相关性研究

分析中国北方汉族白癜风的临床特征及与HLA-DRB1等位基因的相关性,探讨白癜风的病因及发病机制.登记125例白癜风门诊患者的临床资料,利用聚合酶链反应-序列特异引物(PCR-SSP)技术对125例白癜风患者和263例健康人进行等位基因检测,对各项指标进行统计分析.结果,白癜风初发年龄男性与女性无显著差异;家族史阳性患者发病年龄早于家族史阴性患者;伴有自身免疫性疾病平均初发年龄大于不伴自身免疫疾病的白癜风患者;白癜风与HLA-DRB1*070X及HLA-DRB1*1201等位基因有相关性.①有家族史和无家族史白癜风患者在遗传背景上存在差异,HLA-DRB1*070X可能为家族史阳性患者的易感基因.②提示了节段型与非节段型白癜风的发病机制有所不同,泛发性及散发性与自身免疫相关明显.③HLA-DRB1*070X及HLA-DRB1*1201可能是中国北方汉族寻常型白癜风的易感基因或与易感基因相连锁.     

得宝松穴位封闭联合甲氧沙林等药物治疗节段型、限局型白癜风

目的　探索节段型、限局型白癜风新的治疗方案。方法　随机将 60例节段型、限局型白癜风分为两组。A组患者采用内服甲氧沙林片、薄芝片 ,外用甲氧沙林酊剂的治疗方案 ;B组患者采用A组患者所用药物联合得宝松穴位封闭的治疗方案。结果　A组、B组的总有效率分别为 44.44%、75.86% ,二者差异具有显著性 (χ2 =5.79,P <0 .0 5) ;A组、B组的光毒反应发生率分别为 88.3 3 %、73 .3 3 % ,二者差异无显著性 (χ2 =0 .88,P >0 .0 5)。结论　得宝松穴位封闭联合甲氧沙林片 (酊 )、薄芝片是一种治疗节段型、限局型白癜风新的、合理的、值得推广的方案     

寻常型银屑病并发白癜风2例

银屑病和白癜风均为较常见的与自身免疫有关的皮肤疾病。而其合并发生的具体机制目前仍不明了。本文报道2例银屑病伴发白癜风，并对国内外近年来银屑病并发白癜风的文献报道进行了复习，从而进一步探究二者合并发病的可能机制。     

体外培养节段型白癜风样无色素痣皮损黑素细胞及其临床意义

【摘要】 目的 评价负压吸疱表皮黑素细胞培养对节段型白癜风样无色素痣的辅助诊断价值。方法 收集2019年6月至2020年3月于杭州市第三人民医院皮肤科依据Coupe标准临床诊断的8例节段型白癜风样无色素痣患者,进行伍德灯、反射式共聚焦显微镜（RCM）检查,308 nm准分子激光试验,负压吸疱获取表皮黑素细胞并培养,记录结果。结果 8例患者中,6例皮损伍德灯下可见荧光,4例RCM下可见色素环完整性缺失,5例经308 nm准分子激光照射试验后未见复色反应。8例患者体外培养的皮损黑素细胞硫酸亚铁染色均阳性,经消化离心后沉淀呈黄白色,电镜下黑素细胞胞质内可见Ⅰ~ Ⅲ期黑素小体;皮损对侧同一解剖部位正常皮肤黑素细胞沉淀则呈黑色,电镜下黑素细胞胞质内可见Ⅰ~ Ⅳ期黑素小体。结论 负压吸疱表皮黑素细胞培养有助于诊断节段型白癜风样无色素痣。     

节段型白癜风皮损区角朊细胞表达碱性成纤维细胞生长因子的观察

近年体外研究发现角朊细胞(ＫＣ)能产生多种影响黑素细胞(ＭＣ)增殖、黑素形成、形态、分化及其存活的细胞因子[1],其中以碱性成纤维细胞生长因子(ｂＦＧＦ)、内皮素1(ＥＴ1)和干细胞因子(ＳＣＦ)最为重要。Ｈａｌａｂａｎ等的体外研究证实[2],来源于ＫＣ的ｂＦＧＦ是正常ＭＣ的自然生长因子。白癜风的基本病理改变是表皮内的ＭＣ减少或消失,其发生的确切原因仍不清楚。了解ＫＣ所分泌的细胞因子ｂＦＧＦ在不同病期节段型白癜风皮损处ＫＣ的表达状况,为判断ＭＣ自体移植治疗白癜风后白斑区ＭＣ的转归提供一定的实验依据以及进一步认识该病的…     

Autoantibodies against tyrosine hydroxylase in patients with non-segmental (generalised) vitiligo

Vitiligo is an acquired idiopathic hypomelanotic skin disorder characterised by depigmented macules because of loss of cutaneous melanocytes. Although the exact cause of vitiligo remains obscure, evidence suggests that autoimmunity plays a role in the pathogenesis of the disease. Previously, tyrosine hydroxylase (TH) was identified as a putative autoantigen in vitiligo using phage-display technology. In this study, the prevalence of TH antibodies in patients with vitiligo was investigated. A radioimmunoassay (RIA) was used to detect TH antibodies in sera from patients with either non-segmental vitiligo (n=79), segmental vitiligo (n=8) or other autoimmune diseases without concomitant vitiligo (n=91). Sera from healthy individuals (n=28) were also tested. Patients with segmental vitiligo, healthy controls and patients with other autoimmune diseases without concomitant vitiligo were all negative for TH antibody reactivity. Of 79 patients with non-segmental vitiligo, 18 (23%) were positive for TH antibodies in the RIA, and a significant increase in the prevalence of TH antibodies in patients with non-segmental vitiligo was evident when compared with controls (P=0.003). TH antibody prevalence was also significantly elevated in patients with active vitiligo compared to those with stable disease (P=0.009). Overall, the results indicate that TH is an antibody target in non-segmental but not in segmental vitiligo and that TH antibodies appear to be more frequent in patients with active vitiligo.     

系统糖皮质激素治疗进展期白癜风的疗效及相关因素研究

目的:探讨系统糖皮质激素（简称激素）治疗白癜风疾病活动度评分（VIDA）≥ 2分的进展期患者的疗效及相关因素。方法:2018年6月至2019年6月于杭州市第三人民医院皮肤科,收集未经系统性激素治疗、VIDA ≥ 2分、皮损面积<1%的进展期白癜风患者272例,分析皮损面积和型别、VIDA评分、是否具有诱发因素及特殊临床...     

原位末端标记法对白癜风皮肤凋亡细胞的检测

目的：探讨白癜风表皮细胞凋亡与细胞坏死组织形态特点。方法：应用原位末端标记（ＩＳＥＬ）方法。结果：细胞凋亡基本特征是ＤＮＡ断裂。结论：ＤＮＡ断裂是细胞凋亡的主要标志，此可区别于坏死细胞。     

Increased micronucleus frequency in phytohaemagglutinin-stimulated blood cells of patients with vitiligo

Background Vitiligo is a relatively common, acquired pigmentary disorder characterized by areas of depigmented skin resulting from loss of melanocytes in the epidermis. Although several hypotheses have been proposed for the aetiology and pathogenesis of vitiligo, the cause of vitiligo remains unclear. Objective To evaluate spontaneous micronucleus (MN) frequency using the cytokinesis block MN assay to determine damages at the DNA or chromosome level in phytohaemagglutinin (PHA)‐stimulated blood cells of patients with vitiligo and healthy control subjects. Methods Peripheral blood samples were obtained and cultured from 21 patients with vitiligo (mean age: 21.48 ± 9.78 years) and 21 age‐ and sex‐matched healthy control subjects (mean age: 21.52 ± 9.80 years). MN values were scored in binucleated cells obtained from whole‐blood cultures of patients and control subjects. Results MN frequencies (mean ± SD) in PHA‐stimulated blood cells of patients with vitiligo and control subjects were 0.94 ± 0.58 and 0.58 ± 0.32, respectively. Compared with control subjects, MN frequencies of patients with vitiligo were found significantly higher than those of the control subjects (P = 0.012). Conclusion Our results indicate unexpectedly some chromosomal/DNA damage in whole‐blood cultures of patients with vitiligo. We do not know, however, if these chromosome/DNA instabilities observed in the cells of vitiligo patients resulted from the cause or from the consequences of the disorder.     

寻常型进行期白癜风患者白斑区角质形成细胞凋亡的检测

目的探讨进行期白癜风患者白斑区角质形成细胞是否出现凋亡及其凋亡相关蛋白的表达。方法标本分别取自15例进行期白癜风患者白斑中心区及15例正常人对照，均取材于胸腹部。采用末端脱氧核苷酸转移酶介导的dUTP缺口末端标记技术（TUNEL）检测角质形成细胞的凋亡状况，同时采用免疫组化法检测角质形成细胞表达凋亡相关蛋白Bax、Bcl-2和Caspase3的情况。结果TUNEL结果显示：15例白癜风患者白斑区均可见角质形成细胞存在不同程度凋亡，主要位于表皮中层及下层，密集或散在分布；15例正常人对照中仅有1例表皮可见10％极为散在的凋亡的角质形成细胞，两组比较差异有统计学意义（P〈0．01）。免疫组化结果显示：7例白癜风患者白斑区可检测到部分角质形成细胞存在促凋亡相关蛋白Bax的弱表达，而对照组仅有1例TUNEL阳性者可检出Bax的弱表达，即白癜风皮损区角质形成细胞Bax表达较正常对照显著升高，差异有统计学意义（P〈0．01）。所有标本中角质形成细胞均未检测到Bcl-2和Caspase3的表达。结论进行期白癜风白斑区角质形成细胞促凋亡蛋白Bax表达升高，其凋亡增加。     

白癜风患者215例临床分析

目的：总结白癜风患者的临床特点和发病诱因.方法：对确诊的白癜风患者进行问卷设计和调查,用SPSS 11.5软件包进行统计分析.结果：资料完整的215例白癜风患者中男98例,女117例;平均发病年龄18.76±7.43岁;最常见的受累部位为上肢（54.42%）;伴发疾病中以甲状腺疾病最常见（5.58%）;有家族史者占13.02%,有家族史患者发病年龄为14.6±4.31岁,无家族史为25.09±5.45岁（P〈0.001）,两者间有显著性差异;春夏季发病最常见;25.58%的患者可发现诱因,其中常见的诱因包括精神因素 （43.64%）和皮肤损伤（56.36%）.结论：本地区白癜风好发于青少年,有家族史患者发病年龄早于无家族史患者,上肢为最常见的好发部位,可能与精神因素和皮肤外伤、日晒伤有关.     

Melanocyte detachment after skin friction in non lesional skin of patients with generalized vitiligo

BACKGROUND: In vitiligo, melanocytes are gradually lost in depigmented macules of the skin. The disappearance of melanocytes has, however, not been clearly observed and consequently the aetiology of the disease (autoimmune, neural, cytotoxic) is still elusive. The starting point of vitiligo macules is frequently determined by local conditions such as wounds and excoriations, but may also follow minor traumas such as pressure or repeated friction. This prominent feature is often neglected. OBJECTIVES: To clarify the biological consequences of repeated friction on the attachment and survival of melanocytes in non lesional vitiligo skin. METHODS: Light reproducible skin friction was performed for 4 min on the volar forearm of 18 patients with extensive vitiligo and five controls with normal healthy skin. Biopsies from the test area and control skin were taken at 1, 4, 24 and 48 h following friction. Serial sections were examined with standard light microscopy, transmission electron microscopy, histochemistry and immunohistochemistry (dihydroxyphenylalanine, HMB-45, E-cadherin and an early apoptosis marker, M30 cytoDEATH antibody). RESULTS: The observation of sections at 1 and 48 h after friction on vitiligo skin and at all time points in controls revealed no changes. In contrast, in vitiligo skin at 4 and 24 h after friction, several melanocytes had undergone detachment and were found in various suprabasal positions, including the stratum spinosum, granular layer, and within and outside the stratum corneum. CONCLUSIONS: Detachment and transepidermal elimination of melanocytes following minor mechanical trauma in non lesional vitiligo skin is probably the cause of depigmentation occurring in the isomorphic response (Koebner phenomenon). We propose that transepidermal elimination of melanocytes in vitiligo should be regarded as a possible mechanism of chronic loss of pigment cells, perhaps previously damaged by another process.