桉树脑对HaCaT细胞水通道蛋白3和透明质酸表达的影响

目的:探讨桉树脑(Cin)对人永生化角质形成细胞(HaCaT)水通道蛋白(AQP)3及透明质酸(HA)转录和翻译的影响。方法:以倍他米松(BT)为对照,不同浓度Cin(0.01%、0.001%、0.0001%和0.00001%)分别孵育HaCaT细胞36 h后,采用实时荧光定量聚合酶链式反应(q-PCR)检测AQP3 mRNA表达水平,免疫印迹法(western blot)分析AQP3蛋白表达,以及酶联免疫吸附试验(ELISA)检测细胞外基质中HA的含量。结果:与空白对照组相比,qPCR结果示0.01%和0.001%Cin均显著增加了AQP3基因的转录(P0.05);而10μmol/L BT明显下调AQP3基因转录(P0.05);0.01%和0.001%Cin分别与10μmol/L BT共同孵育HaCaT细胞后,均可逆转10μmol/L BT引起的AQP3 m RNA表达下调(P0.05)。Western blot结果表明,与空白对照组比较,0.01%Cin组和0.001%Cin组AQP3蛋白表达均显著增加(P0.05);10μmol/L BT组AQP3蛋白表达明显下调(P0.05)。与空白对照组比较,0.01%Cin+10μmol/L BT组和0.001%Cin+10μmol/L BT组AQP3蛋白表达均下降(P0.05);与10μmol/L BT组比较,AQP3蛋白表达均明显提高(P0.05)。ELISA结果表明,与空白对照组比较,10μmol/L BT组HA表达水平下调(P0.05);与10μmol/L BT组比较,0.01%Cin+10μmol/L BT组和0.001%Cin+10μmol/L BT组HA表达水平均上调(P0.05),但均低于空白对照组(P0.05)。结论:Cin可不同程度逆转BT对角质形成细胞AQP3和HA的抑制作用。     

白芍总苷对HaCaT细胞表达白细胞介素18的影响及相关信号通路的研究北大核心CSCD

目的 观察白芍总苷（TGP）对角质形成细胞（KC）表达白细胞介素（IL）-18的影响,并初步探讨ERK1/2、JNK1/2信号通路在其中的作用.方法 将部分HaCaT细胞分为3个组,即对照组加入0.031％二甲基亚砜的细胞培养液,TGP组分别加入6种不同浓度的TGP（0.5、2.5、12.5、62.5、125.0、312.5 mg/L）,抑制剂组分别加入10μmol/L ERKl/2抑制剂PD98059和JNK1/2抑制剂SP600125预处理2h后,再加入125 mg/LTGP.细胞继续培养48 h.实时反转录（RT）-PCR方法和ELISA方法检测HaCaT细胞IL-18的表达.部分HaCaT细胞分为两组,一组用125 mg/L TGP分别处理15,30,60 min,另一组分别用10μmol/L ERKl/2抑制剂PD98059和JNK1/2抑制剂SP600125预处理后再加入125 mg/L TGP分别处理15,30,60 min.免疫印迹技术观察两组HaCaT细胞ERK1/2、JNK1/2磷酸化水平.结果 TGP在低浓度（0.5、2.5 mg/L）时对HaCaT细胞IL-18mRNA和蛋白的表达有促进作用,62.5～125.0 mg/L时可抑制IL-18 mRNA和蛋白的表达,125 mg/L TGP抑制作用最强.TGP（125 mg/L）作用15 min后磷酸化ERKl/2蛋白表达达到高峰,表达水平为0.448±0.018,与对照组（0.204±0.005）比较,差异有统计学意义（P＜0.01）;30 min后表达水平降低至0.213±0.005,60 min后为0.217±0.005,与对照组相比差异无统计学意义（均P＞0.05）.PD98059预处理组磷酸化ERK1/2表达水平为0.237±0.010,与单独125 mg/L TGP给药组相比,差异有统计学意义（P＜0.01）.125 mg/L TGP对JNK蛋白的磷酸化作用不明显,各组相比差异无统计学意义（P＞0.05）.结论 TGP可抑制IL-18 mRNA和蛋白的表达,ERK1/2信号途径可能介导这一抑制作用.     

金黄色葡萄球菌肽聚糖对HaCaT细胞Toll样受体2和4的影响

目的观察金黄色葡萄球菌胞壁成分肽聚糖对HaCaT细胞Toll样受体(TLR)2和TLR4表达的影响。方法用不同浓度的肽聚糖(10μg/mL、30μg/mL、100μg/mL)处理培养的HaCaT细胞,用RT-PCR检测TLR2 mRNA和TLR4 mRNA含量变化,用流式细胞术检测HaCaT细胞表面TLR2和TLR4蛋白的表达。结果30μg/mL肽聚糖处理组HaCaT细胞的TLR2 mRNA和TLR4 mRNA相对表达水平分别为0.826±0.06和0.801±0.04,明显高于未处理的对照组的TLR2 mRNA(0.433±0.04,P<0.05)和TLR4 mRNA(0.351±0.05,P<0.01)。各浓度的肽聚糖处理HaCaT细胞24h,及100μg/mL浓度的肽聚糖处理HaCaT细胞12h、24h、36h和48h时,及100μg/mL肽聚糖处理24h后,HaCaT细胞表面TLR2和TLR4蛋白表达量均最高。结论肽聚糖可以上调HaCaT细胞TLR2和TLR4的mRNA和蛋白的表达。     

表皮生长因子对HaCaT细胞miR-21/PCD4的表达研究

目的 探讨表皮生长因子(EGF)对角质形成细胞(HaCaT细胞)miR-21/PDCD4表达的调控作用.方法 聚合酶链反应(PCR)法检测不同浓度EGF(0、5、15、25 ng/mL)对HaCaT细胞miR-21表达的影响;蛋白免疫印迹法检测不同浓度EGF(0、5、15、25 ng/mL)对HaCaT细胞PDCD4蛋白表达的影响.结果 EGF浓度依赖性促进HaCaT细胞miR-21的表达,与对照组比较,5 ng/mL EGF组、15 ng/mL EGF组、25 ng/mL EGF组HaCaT细胞miR-21表达分别为1.46±0.11、2.44±0.05、3.19±0.08(P＜0.05);EGF浓度依赖性抑制HaCaT细胞HaCaT细胞PDCD4表达,跟对照组比较,25 ng/mL EGF组能显著抑制HaCaT细胞PDCD4表达.结论 EGF促进HaCaT细胞miR-21的表达,抑制HaCaT细胞PDCD4的表达水平.     

白芍总苷对角质形成细胞增殖及血管内皮生长因子和白介素-23表达的影响

目的 研究白芍总苷（TGP）对角质形成细胞增殖和分泌血管内皮生长因子（VEGF）和白介素（IL）-23的影响,探讨可能涉及的信号转导通路。方法 不同浓度TGP作用于体外培养的HaCaT细胞株,噻唑蓝（MTT）法观察TGP对HaCaT细胞增殖活性的影响。将HaCaT细胞分为三组,即对照组不加任何刺激因素,TGP组分别加入6种不同浓度的TGP,SB203580组在加入10 mol/L SB203580预处理2 h后加入125 mg/L TGP。实时定量PCR（RT-PCR）方法和ELISA方法检测TGP对HaCaT细胞VEGF和IL-23表达的影响;免疫印迹技术观察TGP作用于HaCaT细胞后p38的磷酸化及SB203580对p38磷酸化的影响。结果 TGP在低浓度（0.5、2.5 mg/L）时对HaCaT细胞增殖有促进作用,浓度≥12.5 mg/L时反而对细胞的增殖有抑制作用,至125 mg/L时抑制作用最强。TGP在低浓度（0.5、2.5 mg/L）时对HaCaT细胞VEGF和IL-23 mRNA和蛋白的表达有促进作用,12.5 ～ 125 mg/L时内可抑制HaCaT细胞VEGF mRNA和蛋白的表达,62.5 ～ 125 mg/L时可抑制IL-23 mRNA和蛋白的表达。TGP可时间依赖性地诱导HaCaT细胞p38的磷酸化,磷酸化p38 蛋白于125 mg/L TGP作用5 min后达到高峰,表达水平为0.3314 ± 0.0245,10 min后减弱至0.2173 ± 0.0189,但均高于对照组水平;30 min后表达水平降为0.1664 ± 0.0201;SB203580可减弱其作用,SB203580预处理组磷酸化p-p38 表达水平为0.1529 ± 0.0147。结论 TGP可抑制HaCaT细胞的增殖及VEGF和IL-23 mRNA和蛋白的表达,p38MAPK信号途径可能介导其抑制作用。     

葡萄籽提取物原花青素对UVA照射HaCaT细胞表达AQP3、Caspase-14和BH mRNA的影响

目的通过研究UVA照射前后及不同浓度葡萄籽提取物原花青素(grape-seed proanthocyanins extract,GSPE)干预的HaCaT细胞表达水通道蛋白3(AQP3)、半胱氨酸蛋白酶14(Caspase-14)和博来霉素水解酶(BH) mRNA的变化,探索GSPE是否能够修复长波紫外线引起的皮肤屏障损伤。方法实验分为五个组:空白对照组、单纯光照组(选取UVA的能量密度为25 J/cm~2)、照光联合不同浓度GSPE干预组(10、50、100μg/mL),采用RT-PCR法分别检测各组AQP3、Caspase-14和BH mRNA的表达量。结果 25 J/cm~2的UVA可以提高AQP3 mRNA表达量。GSPE药物浓度大于10μg/mL时,随药物浓度增加,可以逐渐降低光照后高表达的基因水平,并呈浓度依赖性。UVA可下调BH和Caspase-14 mRNA表达量,而GSPE可以提高光照后降低的基因表达量,且呈浓度依赖性,逐渐使其恢复正常。结论合适浓度的GSPE可以调节UVA对HaCaT细胞AQP3、Caspase-14和BH mRNA表达的影响,并使其趋于正常,推测其具有修复长波紫外线引起的皮肤屏障功能损伤的作用。     

角质形成细胞生长因子受体反义寡核苷酸对HaCaT细胞增殖的影响

目的 研究角质形成细胞生长因子受体反义寡核苷酸(KGFR ASODN)对角质形成细胞生长因子(KGF)促HaCaT细胞增殖效应的抑制作用。方法 HaCaT细胞体外培养,采用绘制生长曲线、MTT实验、集落形成实验研究KGFR ASODN对KGF促增殖效应的抑制作用。采用流式细胞仪研究KGFR ASODN对KGFR表达的影响。结果 反义序列1和反义序列2均可抑制HaCaT细胞的生长速度(P<0.05)。高于0.25μg/mL质量浓度反义序列1和反义序列2均可抑制KGF介导的HaCaT细胞增殖(P<0.05)。高于0.25μg/mL质量浓度反义序列1和反义序列2都可抑制KGF诱导的HaCaT细胞集落形成(P<0.05)。HaCaT细胞存在KGFR基础表达,高于0.25μg/mL质量浓度反义序列1和反义序列2均可抑制HaCaT细胞KGFR表达(P<0.05),抑制率呈浓度依赖性,正义序列核苷酸(S-ODN)对KGFR表达无抑制作用(P>0.05)。结论 KGFR ASODN可有效抑制KGFR表达,进而抑制KGF介导的HaCaT细胞过度增殖。     

表皮生长因子对HaCaT细胞miR-21/PDCD4的表达研究

目的探讨表皮生长因子(EGF)对角质形成细胞(HaCaT细胞)miR-21/PDCD4表达的调控作用。方法聚合酶链反应(PCR)法检测不同浓度EGF(0、5、15、25 ng/mL)对HaCaT细胞miR-21表达的影响;蛋白免疫印迹法检测不同浓度EGF(0、5、15、25 ng/mL)对HaCaT细胞PDCD4蛋白表达的影响。结果 EGF浓度依赖性促进HaCaT细胞miR-21的表达,与对照组比较,5 ng/mL EGF组、15 ng/mL EGF组、25 ng/mL EGF组HaCaT细胞miR-21表达分别为1.46±0.11、2.44±0.05、3.19±0.08(P0.05);EGF浓度依赖性抑制HaCaT细胞HaCaT细胞PDCD4表达,跟对照组比较,25 ng/mL EGF组能显著抑制HaCaT细胞PDCD4表达。结论 EGF促进HaCaT细胞miR-21的表达,抑制HaCaT细胞PDCD4的表达水平。     

Dermatology in private practice in France in 2000

OBJECTIVES: Data regarding French dermatological practice are scarce. Our objective was to identify the skin disorders most commonly diagnosed by office-based dermatologists. We also documented the severity of these skin disorders, as reflected by the repercussions on patient's everyday life, and the way physicians managed patients. DESIGN: We carried out a one-day survey of visits to a randomly selected sample of 900 French office-based dermatologists. The randomization was stratified according to the five French different dialing area codes. RESULTS: Office-based dermatologists saw 6411 patients with 7839 skin disorders during the survey. The daily number of visits to French dermatologists was estimated at 47 000 and the annual number between 12 and 14 millions. Office-based dermatologists mostly managed warts, acne, nevus, dermatitis, malignancies and pre-malignancies, fungal infection and psoriasis. Repercussions on patients'everyday life were assessed by physicians as important or very important in 28 p. 100 of cases. Half of the patients received topical treatment, 20.5 p. 100 a systemic drug and 40 p. 100 a minor surgical procedure (including cryotherapy). CONCLUSION: Although dermatologists frequently see benign skin disorders such as warts or nevus, more severe diseases represent an important part of their activity.     

Decrease in neutrophils observed in vivo in psoriatics after PUVA therapy

Summary In 56 patients leucocyte and differential counts were done before and at weekly intervals during PUVA treatment of chronic recalcitrant psoriasis. A statistical significant (P<0.01) decrease in the percentage of neutrophils was observed during the first week of the PUVA therapy. This observation could be closely related to the clinical clearing of psoriasis (P=0.02).The effect of PUVA therapy in psoriasis may be due to a decrease in the number of immunocompetent neutrophils demonstrated in psoriatic lesions.     

37例麻风患者的家庭集聚性调查分析

目的:明确麻风家庭感染者的发病情况。方法:利用全国麻风病防治信息管理系统(LEPMIS)数据库,描述性分析麻风家庭感染者的发病情况。结果:符合条件的共17户家庭37例麻风患者,其中多菌型患者33例,少菌型4例;密切接触5年内发病4例(10.81%),5~10年发病者11例(29.73%),超过10年发病者13例(35.14%),不明时间者9例(24.32%);其中因父母子女关系感染17例(45.95%)、祖孙关系5例(13.51%)、兄弟关系2例(5.41%)。结论:多菌型麻风是家庭传染者的主要传染源。     

Progress in Malassezia Research in Korea

Yeasts of the genus Malassezia are part of the normal flora of human skin. However, they are also associated with various skin diseases. Since the introduction of Malassezia to the Korean Dermatologic Society two decades ago, remarkable progress has been made in our knowledge of this genus. In this paper, we review recent developments in Malassezia research, including taxonomy and methods for species identification, recent genome analyses, Malassezia species distribution in healthy conditions and in specific skin diseases, trials investigating the mechanisms underlying Malassezia-related diseases, as well as therapeutic options. This review will enhance our understanding of Malassezia yeasts and related skin diseases in Korea.     

Hard cornification in reptilian epidermis in comparison to cornification in mammalian epidermis

The structure of reptilian hard (beta)-keratins, their nucleotide and amino acid sequence, and the organization of their genes are presented. These 13-19 kDa proteins are basic, rich in glycine, proline and serine, and different from cytokeratins. Their mRNAs are expressed in beta-cells. The central part of beta-keratins (this region has been previously termed 'core-box' and is peculiar of all sauropsid proteins) is composed of two beta-folded regions and shows a high identity with avian beta-keratins. This central part present in all beta-keratins, including feather keratins, is the site of polymerization to build the framework of beta-keratin filaments. Beta-keratins appear cytokeratin-associated proteins. Their central region might have originated in an ancestral glycine-rich protein present in stem reptiles from which beta-keratins evolved and diversified into reptiles and birds. Stem reptiles of the Carboniferous period might have possessed glycine-rich proteins derived from exons/domains corresponding to the variable, glycine-rich region of cytokeratins. Beta-keratins might have derived from a gene coding for small glycine-rich keratin-associated proteins. The glycine-rich regions evolved differently in the lineage leading to modern reptiles and birds versus that leading to mammals. In the reptilian lineage some amino acid regions produced by point mutations and amino acid changes might have given rise to originate the central beta-pleated region. The latter allowed the formation of filamentous proteins (beta-keratins) associated with intermediate filament keratins and replaced them in beta-keratin cells. In the mammalian lineage no beta-pleated region was generated in their matrix proteins, the glycine-rich keratin-associated proteins. The latter evolved as glycine-tyrosine-rich, sulphur-rich, and ultra-sulphur-rich proteins that are used for building hairs, horns and nails.     

Experimental approaches to lymphocyte migration in dermatology in vitro and in vivo

Lymphocyte trafficking through the dermal compartment is part of the physiological surveillance process of the adaptive immune system. On the other hand, persistent or recurrent lymphocyte infiltrates are hallmarks of both types of chronic inflammatory skin diseases, Th1-type such as psoriasis or Th2/allergic-type like atopic dermatitis. A better understanding of the mechanisms underlying lymphocyte movements is one of the key prerequisites for developing more effective therapies. In this review, we introduce a range of simple-to-sophisticated experimental in vitro and in vivo approaches to analyze lymphocyte migration. These methods start from static in vitro adhesion and chemotaxis assays, include dynamic endothelial flow chamber, intravital dual photon, and transcutaneous live-video microscopy, and finally encompass specific genetically deficient or engineered animal models. Discussing pros and cons of these assay systems hopefully generates both state-of-the-art knowledge about the factors involved in most common chronic skin diseases as well as an improved understanding of the limitations and chances of new biologic pharmaceuticals that are currently introduced into clinical practice.     

Dermatoses in cement workers in southern Taiwan

Construction workers are known to have occupational dermatoses. The prevalence of such dermatoses was unknown in Taiwanese construction workers. The objective of this study was to determine the work exposure, prevalence of skin manifestations, and sensitivity to common contact allergens in cement workers of southern Taiwan. A total of 1147 current regular cement workers were telephone-interviewed about skin problems during the past 12 months, work exposure, and personal protection. Among those interviewed, 166 were examined and patch tested with common contact allergens. A high % of cement workers reported skin problems in the past 12 months. More men (13.9%) reported skin problems possibly related to work than women (5.4%). Prevalence was associated with lower use of gloves, duration of work as cement worker, and more time in jobs involving direct manual handling of cement, especially tiling. A high % of dermatitis was noted in the 166 workers examined, which correlated with reported skin problems. On patch testing, construction workers had a high frequency of sensitivity to chromate. Sensitivity to chromate or cobalt was associated with reported skin problems, or dorsal hand dermatitis on examination. These workers'dermatitis was under-diagnosed and inadequately managed. It is concluded that cement workers in southern Taiwan had a high prevalence of skin problems related to cement use. Protective measures, work practice, and physician education should be improved to prevent or manage such problems.