甘草苷对黄褐斑小鼠模型SOD,MDA,NO含量及黑素瘤A375细胞增殖的影响

<正>60只昆明种雌性小鼠,按体重随机分为空白对照组、模型组、甘草苷(LQ)低剂量组(15mg/kg.d),LQ中剂量组(30mg/kg.d),LQ高剂量组(60mg/kg.d)和维生素C组。空白对照组肌肉注射生理盐水,其余各组肌肉注射黄体酮加紫外线照射法致黄褐斑小鼠模型。各组分别按剂量灌胃给药30d后,检测小鼠血清和皮肤中超氧化物歧化酶(SOD)、丙二醛(MDA)活性和一氧化氮(NO)含量。用四甲基偶氮唑蓝比色法(MTT)法     

乌司他丁对内毒素休克大鼠器官功能的保护作用

目的 探讨蛋白酶抑制剂对于内毒素所致大鼠休克时器官功能的保护作用及其可能的机制.方法 健康雄性Wistar大鼠共48只,分为两部分,每一部分24只,分为三组.其中第一部分设生理盐水对照组(C组,n=6);内毒素组(E组,n=10),尾静脉注射内毒素5 mg/kg(lipopolysaccharide,LPS O55B5);蛋白酶抑制剂组(U组,n=8),尾静脉给与LPS 5 mg/kg,同时分别给予腹腔注射蛋白酶抑制剂乌司他丁10万U/kg.第二部分三组给药方法相同,不取组织和血标本,仅观察2 h血压及24 h死亡率.第一部分三组动物2 h采集血、尿标本及肺组织,分别测定肺组织湿/干重比值及髓过氧化物酶(MPO)、脂质过氧化代谢产物水平(丙二醛MDA和共轭二烯C-diense),测定动脉血气、血乳酸(Lactic acid)和内皮素(ET)水平、血浆肌酐(SCr)及尿N-乙酰氨基葡萄糖苷酶(NAG)水平.结果 U组血压低于C组[(99.16±6.91 vs 120.5±3.41)mmHg](P＜0.001)而高于E组[(99.16±6.91 vs 75±4.16)mmHg](P＜0.001);血气分析示PaO2 U组高于E组[(112.8±4.83 vs 82.8±4.03)mmHg](P＜0.01);肺湿/干重U组明显低于E组(5.0±0.05vs5.41±0.06)(P＜0.001);肺组织MPO活性,U组明显低于E组[(119.40±11.32 vs 152.90±8.41)u/g.w.w](P＜0.001),肺组织脂质过氧化物水平U组较E组明显降低[(MDA 51.26±5.56 vs 73.95±4.62;C-diense 7.59±0.84 vs 10.96±0.81)nmol/g.w.w](P＜0.01);尿NAG与血浆肌酐水平U组明显低于E组分别为[(122.56±14.33 vs 337.67±16.52)IU/L及(56.21±4.77 vs 115.67±5.95)μmol/L](P＜0.001);血浆乳酸水平U组较E组降低[(4.03±0.11vs 5.90±0.14)mmol/L](P＜0.001);血浆内皮素水平U组较E组降低[(633.27±93.27 vs 948.23±103.45)pg/ml](P＜0.05),U组大鼠死亡率有下降趋势.结论 蛋白酶抑制剂乌司他丁对内毒素所致休克大鼠的多器官功能损害具有一定的保护作用.     

青海黑果枸杞原花青素对小鼠光老化皮肤的保护作用

目的研究黑果枸杞原花青素对紫外线照射(UV)引起的小鼠皮肤光老化的保护作用。方法 SPF级昆明小鼠50只随机分成5组:正常对照组、模型组、阳性对照组(维生素E组)、原花青素低剂量组、原花青素高剂量组。除了正常对照组,其余组每日颈背部皮下注射5%D-半乳糖(10 ml/kg),同时进行UV[长波紫外线(UVA)+中波紫外线(UVB)]照射。造模第11天开始,正常对照组、模型组每日给予生理盐水灌胃,原花青素低剂量组、原花青素高剂量组每日按剂量[50 mg/(kg·d)和100 mg/(kg·d)]灌胃黑果枸杞原花青素,阳性对照组每只给予维生素E滴剂0.5 ml,连续用药30 d。肉眼观察各组小鼠皮肤外观及皮下血管变化;以酶生化法测定皮肤组织活性氧(ROS)、过氧化氢酶(CAT)活力,酶联免疫吸附试验(ELISA)测定皮肤组织Bax、Bcl-2及血清中白细胞介素(IL)-1、IL-6、肿瘤坏死因子(TNF)-α的含量。结果与正常对照组相比,模型组皮肤组织中Bax、IL-1、IL-6、TNF-α含量显著上升,ROS、CAT活力,Bcl-2的含量下降(P0.05);与模型组比较,黑果枸杞原花青素低、高剂量组皮肤组织中Bax、IL-1、IL-6、TNF-α含量的表达水平显著下降,ROS、CAT活力,Bcl-2含量上升,差异具有统计学意义(P0.05)。模型组皮肤颜色变暗、粗糙肥厚,出现皱纹,并伴有脱屑;黑果枸杞原花青素组皮肤外观较模型组明显改善。结论黑果枸杞原花青素对UV引起的小鼠皮肤光老化具有保护作用,其机制可能与黑果枸杞原花青素抑制细胞内活性氧产生,降低线粒体通透性,抑制细胞色素C释放,抑制细胞凋亡,减轻炎症反应有关。     

与黑素结合的羟氯喹对长波紫外线诱导HaCaT细胞死亡的光保护机制研究

目的:观察与黑素结合的羟氯喹(HCQ)对细胞内活性氧基(ROS)的清除以及对长波紫外线(UVA)诱导人角质形成细胞(HaCaT细胞)凋亡和坏死的保护.方法:自一株产黑素的嗜麦芽假单胞菌AT18的培养液中分离纯化获得细菌衍生黑素,在体外与不同浓度HCQ结合后处理培养的HaCaT细胞,随后给予半数致死剂量(30 J/cm2)的UVA照射.照射12 h后,以四甲基偶氮唑蓝(MTT)比色法测定细胞存活率;碘化丙啶(PI)染色结合流式细胞仪检测细胞凋亡率;采用二氯荧光素二酯(DCFH-DA)标记法测定细胞内ROS水平.结果:低浓度(10、50 μmol/L)HCQ与黑素结合后能明显保护HaCaT细胞抵抗半数致死剂量(30 J/cm2)UVA照射,尤其是50 μmol/L HCQ 50 mg/L黑素处理组细胞存活率较单纯黑素和单纯HCQ组显著增高(P＜0.05);相反,用高浓度(250 μmol/L)HCQ与黑素结合处理细胞,细胞存活率较单纯黑素和单纯HCQ组减低.与单纯黑素和单纯HCQ组相比,10 μmol/L HCQ与黑素结合还能显著提高细胞内ROS的清除能力和抑制UVA'诱导HaCaT细胞凋亡.结论:低浓度HCQ与黑素结合能协同增强黑素对细胞内ROS的清除,抑制UVA诱导的皮肤细胞凋亡,这些作用很可能关系到治疗皮肤型红斑狼疮时抗疟药的抗光敏机制.     

乌司他丁对急性心肌梗死心肺复苏术后多脏器功能保护作用的临床研究

目的探讨乌司他丁对急性心肌梗死患者心肺复苏术（CPR）后多脏器功能的保护作用。方法对ICU急性心肌梗死CPR后的50例危重患者，随机分为乌司他丁干预组（25例）和综合治疗对照组（25例），对两组的临床和实验室指标进行观察比较。结果两组患者的年龄和CPR后的APACHE-Ⅱ评分比较，差异无统计学意义（P〉0．05），有可比性。干预组机械通气时间为（257．81±97．65）h，显著短于对照组[（425．33±81．57）h，P=0．000]；因发生多器官功能衰竭而死亡患者干预组6／25（24％）例，对照组15／25（60％）例，两组间比较差异均有统计学意义（P〈0．05）；干预组治疗第1天的肌酸激酶同工酶值为（185．13±13．36）U／L，与对照组（183．12±12．57）U／L比较，差异无统计学意义（P=0．5863）；干预组治疗第3和第5天的肌酸激酶同工酶值为（110.56±12．14）U／L和（39．73±10．37）U/L.均显著低于对照组（167．35±11．49）U／L和（73．52±10．18）U/L.P〈0．05）；治疗前后MODS评分结果显示对照组无明显改善，干预组明显降低，治疗第5天干预组MODS评分为（1．59±1．36），明显低于对照组（2．96±0．23，P〈0．05）。结论乌司他丁对急性心肌梗死患者能改善其心肌酶学和降低心律失常的发生率，乌司他丁对急性心肌梗死CPR后多脏器功能有保护作用。     

细菌衍生黑素对长波紫外线诱导人皮肤成纤维细胞凋亡和坏死的保护

目的:探讨细菌衍生黑素(b-melanin)对长波紫外线(UVA)诱导人成纤维细胞凋亡和坏死产生的光保护作用,为今后将其用作皮肤光保护剂提供依据.方法:正常人成纤维细胞(NL-FB)和着色性干皮病患者成纤维细胞(XP-FB)经UVA照射12 h后,以四甲基偶氮唑蓝(MTT)法检测细胞存活率,Hoechst33258染色法观察早期凋亡细胞核形态学变化.二氯荧光素二酯(DCFH-DA)标记法测定细胞内活性氧基(ROS)水平.结果:UVA照射诱导细胞的半数致死剂量,XP-FB大约为30 J/cm2,而NL-FB＞40 J/cm2.为了观察不同浓度(0,25,50,100,200,400,和800 μg/mL)的b-melanin是否对细胞存在光保护作用,给予半数致死剂量(30 J/cm2)UVA照射后,经100～400 μg/mL b-melanin处理的XP-FB的细胞存活率均较未处理组明显增高(P＜0.01),而NL-FB的细胞存活率变化不明显.细胞内ROS测定结果显示100、400 μg/mL的b-melanin能明显清除UVA诱导产生的ROS.100 μg/mL b-melanin即能阻止非致死剂量(16 J/cm2)UVA照射诱导的早期凋亡细胞核改变.结论:b-melanin能对UVA诱导人成纤维细胞凋亡和坏死提供有效的光保护,这一作用很可能关系到b-melanin对ROS的清除.本研究还首次提出核酸切除修复机制缺陷的XP-FB是一敏感的可用于测试UVA光损伤作用的体外细胞模型.     

自拟固胎煎对复发性流产小鼠外周血IL-6、IL-10、IL-17及TGF-β1的影响

目的观察自拟固胎煎对复发性流产模型小鼠外周血白细胞介素-6(IL-6)、白细胞介素-10(IL-10)、白细胞介素-17(IL-17)、转化生长因子-β1(TGF-β1)表达及对妊娠结局的影响。方法选取SPF级CBA/J雌鼠(n=48)、BALB/C雄鼠(n=4)、DBA/2雄鼠(n=20)作为研究对象。采用CBA/J雌鼠×BALB/c雄鼠建立正常妊娠模型,以CBA/J雌鼠×DBA/2雄鼠建立复发性流产模型,复发性流产模型孕鼠随机分为中药组、地屈孕酮组和复发性流产组,且将中药组分为高剂量亚组、中剂量亚组、低剂量亚组,正常妊娠组和复发性流产组给予生理盐水,中药高剂量亚组、中剂量亚组、低剂量亚组依次给予28.5g/kg、14.25g/kg、7.125g/kg自拟固胎煎汤剂,地屈孕酮组给予3.0mg/kg地屈孕酮溶液,连续灌胃2周,每天1次。给药结束后处死并解剖小鼠,观察子宫胚胎数量并计算胚胎丢失率,酶联免疫吸附试验(ELISA)法检测外周血IL-6、IL-10、IL-17及TGF-β1的表达水平。结果与复发性流产组比较,中药中剂量亚组、高剂量亚组和地屈孕酮组胚胎丢失率均显著降低,差异具有统计学意义(P0.05);与复发性流产组比较,中药中剂量亚组、高剂量亚组和地屈孕酮组的IL-6和IL-17的表达均显著降低,IL-10和TGF-β1的表达均显著升高,差异具有统计学意义(P0.05)。结论自拟固胎煎可能通过上调IL-10、TGF-β1的表达水平,下调IL-6、IL-17的表达水平,诱导免疫耐受,减少胚胎丢失率,从而改善妊娠结局。     

姜黄素基于Toll样受体-核因子-κB信号通路对子宫内膜炎大鼠炎性反应的影响

目的 分析姜黄素基于Toll样受体-核因子-κB(TLRs-NF-κB)信号通路对子宫内膜炎大鼠炎性反应的影响。方法 选取43只SPF级SD雌性大鼠,其中10只大鼠作为空白组,另外33只大鼠建立子宫内膜炎模型,将建模成功的30只大鼠分为模型组、低剂量组、高剂量组,各10只。空白组大鼠、模型组大鼠用同等剂量的无菌生理盐水灌胃,低剂量组大鼠给予50 mg/kg姜黄素灌胃,高剂量组大鼠给予100 mg/kg姜黄素灌胃。比较各组的氧化应激反应指标、炎性因子水平及TLRs-NF-κB信号通路蛋白表达量。结果 与空白组相比,模型组、低剂量组、高剂量组的丙二醛(MDA)、白介素-2(IL-2)、白介素-8(IL-8)、白介素-1(IL-1)水平及TLR4、TLR2、NF-κB p65表达量升高,超氧化物歧化酶(SOD)、环氧合酶-2(COX-2)水平降低,差异具有统计学意义(P<0.05);与模型组相比,低剂量组和高剂量组的MDA、IL-2、IL-8、IL-1水平及TLR4、TLR2、NF-κB p65表达量降低,SOD、COX-2水平升高,差异具有统计学意义(P<0.05);与低剂量组相比,高剂量组的MDA、IL-2、IL-8、IL-1水平及TLR4、TLR2、NF-κB p65表达量降低,SOD、COX-2水平升高,差异具有统计学意义(P<0.05)。结论 姜黄素可抑制子宫内膜炎大鼠机体内炎性因子表达,调控子宫内环境平衡,其机制可能与抑制TLRs-NF-κB信号通路、减少蛋白凋亡有关。     

长波紫外线照射对人皮肤成纤维细胞表达一氧化氮合成酶和一氧化氮的影响

目的 通过检测UVA照射后人皮肤成纤维细胞的细胞形态及诱导型一氧化氮合成酶(iNOS) 和一氧化氮(NO)表达,初步探讨UVA引起成纤维细胞光损伤的发病机制.方法 以1J/cm2,5J/cm2和10J/cm2 UVA分别照射人成纤维细胞后24h,用逆转录聚合酶链反应 (RT-PCR )和化学细胞免疫方法检测iNOSmRNA和蛋白表达情况,Griess 法检测NO的含量,MTT试验检测细胞增殖能力,倒置显微镜和HE染色观察细胞形态变化.结果 10J/cm2UVA照射人成纤维细胞后24h出现皱缩最为明显,其细胞增殖能力(MTT)(OD值,0.2472±0.0328)与其他三组比较明显减退(P均<0.05),且iNOSmRNA(1.0568±0.0778)和iNOS蛋白表达强于1J/cm2和5J/cm2(P均<0.05),同时NO的含量(51.11±2.2760)nmol/mL高于1J/cm2的(32.44±3.1620)nmol/mL和5J/cm2的(41.32±2.4460)nmol/mL(P均<0.05).结论 人成纤维细胞iNOS和NO的表达与UVA照射呈剂量依赖.成纤维细胞的光损伤与iNOS和NO产生有关.     

红色毛癣菌病患者血清NO与iNOS表达的实验研究

目的: 测定红色毛癣菌病患者血清中一氧化氮(NO)、诱生型一氧化氮合酶(iNOS)的表达.方法: 应用硝酸还原酶法和分光光度法检测30例手足癣,35例甲真菌病患者和20名正常对照者血清中NO、iNOS的水平.结果: 与正常对照组相比较,红色毛癣菌病患者外周血NO及iNOS血清水平明显降低(P＜0.01).手足癣组与甲真菌病组之间NO及iNOS相互比较,差异无统计学意义(P＞0.05).结论: 红色毛癣菌病患者机体的免疫防御机制不能有效发挥其抗真菌效应,这可能是导致感染易反复和病程迁延的原因之一.     

Occupational allergic contact dermatitis caused by decorative plants

12 cases of occupational allergic contact dermatitis caused by decorative plants were diagnosed in a 14-year period. The patients were middle-aged, and their average exposure time was 13 years. The plant families and plants causing occupational contact dermatitis were Compositae (5 patients: chrysanthemum, elecampane, gerbera, feverfew), Alstroemeriaceae (5 patients, Alstroemeria ), Liliaceae (4 patients; tulip, hyacinth). Amaryllidaceae (2 patients: narcissus) and Caryophyllaceae (2 patients; carnation, cauzeflower). The known chemical allergens causing dermatitis were tuliposide-A and sesquiterepene lactones, such as alantolactones and parthenolide, in the Liliaceae and Compositae families. 7 of the 12 patients were able to continue their work; 5 were not because of severe relapses of skin symptoms. The plant allergen and extract series currently available are of great help in the diagnosis.     

Deutliche Einschränkung der Lebensqualität bei Patienten mit Pemphigus vulgaris: Ergebnisse der deutschen Bullous Skin Disease (BSD)‐Studiengruppe

Background: Pemphigus vulgaris is a potentially life‐threatening autoimmune disorder of the skin and mucous membranes characterized by antibodies against epidermal adhesion molecules. Clinically characteristic are painful chronic blisters or erosions of mucous membranes and skin. There are no published studies on the impact o this disease on quality of life. Patients and methods: This registration was performed within the scope of the German BSD (Bullous Skin Disease) study group, from November 1997 until January 2002. A total of 36 patients with the first diagnosis of pemphigus vulgaris were registered at the university hospitals of Dresden, Erlangen, Kiel, Mannheim, München and Würzburg. Thirty of the 36 (83 %) patients participated in the quality of life questionnaire utilizing the German version of ‘Dermatology Life Quality Index’ (DLQI) provided by A. Y. Finlay. The DLQI varies from 0 to 30 with an increased DLQI score indicating a decrease in quality of quality. Results: The overall DLQI total score of 10 ± 6,7 in the investigated pemphigus patients was significantly increased in comparison to other skin diseases. Conclusions: These results suggest that the DLQI can be a very useful additional outcome criteria for clinical studies with pemphigus vulgaris and in the treatment of these patients.     

Mutations in mevalonate pathway genes in patients with familial or sporadic porokeratosis

Porokeratosis comprises heterogeneous keratinization disorders that are characterized by one or more atrophic patches surrounded by a ridge‐like cornoid lamella. In this study, we evaluated seven families affected by porokeratosis and five sporadic patients of the disease in a Chinese population. We performed Sanger sequencing of exons and flanking intron–exon boundaries of mevalonate pathway genes (MVD, MVK, PMVK and FDPS) and of SLC17A9. In five familial and three sporadic patients, we detected six variations, including four novel mutations (MVD c.1A>G; p.Met1?, c.916G>A; p.Ala306Thr, c.1013+1G>A, and PMVK c.65A>G; p.Lys22Arg) and two recurrent mutations (MVD c.746T>C; p.Phe249Ser, and MVK c.1028T>C; p.Leu343Pro). We then applied I‐TASSER and iGEMDOCK to assess these variants for probable functional impacts. The findings of this study extend the mutation spectrum of porokeratosis and provide further evidence for the genetic basis of this disease.     

[Translated article] Pruritus in Dermatology: Part 1—General Concepts and Pruritogens

Pruritus is the most common symptom of dermatologic and systemic diseases. The diagnosis of pruritus is clinical, although additional tests may be necessary to identify or confirm the cause. Translational medicine has led to the discovery of new mediators of itch, or pruritogens, as well as new receptors. Knowing how to properly recognize the main pathway that mediates itch in each patient is the key to successful treatment. Although the histaminergic pathway predominates in conditions like urticaria or drug-induced pruritus, it is the nonhistaminergic pathway that predominates in nearly all other skin diseases covered in this review. Part 1 of this 2-part review discusses the classification of pruritus, additional testing, the pathophysiology of itch and the pruritogens implicated (including cytokines and other molecules), and central sensitization to itch.     

Anästhesieverfahren in der Dermatologie

Zusammenfassung Die Verfahren der Lokalanästhesie sind integraler Bestandteil der operativen Dermatologie. Sie gewährleisten eine effiziente und sichere Analgesie in umschriebenen Haut- und Weichteilregionen und ermöglichen, einen sonst schmerzhaften diagnostischen oder therapeutischen Eingriff bei erhaltenem Bewusstsein zu tolerieren. Einzelne Methoden der Applikation sind "konkurrenzlos", wie die topische Applikation von EMLA^® oder die Kryoanästhesie, andere bieten alternative Optionen zur Allgemeinanästhesie. Die Tumeszenzlokalanästhesie wurde—jenseits der kosmetischen Liposuktion—zu einer effizienten Anästhesieform für größere Operationen bei Tumoren der Haut, plastische Rekonstruktionen und in der Phlebochirurgie weiterentwickelt. Die Wahl des Verfahrens im Einzelfall wird vom Alter, der Kooperationsfähigkeit und der Komorbidität des Patienten bestimmt. Für Infiltrationsanästhesien werden heute vorwiegend Lokalanästhetika vom Amidtyp eingesetzt. Fundierte Kenntnisse über die Anatomie der sensiblen Nerven sind Voraussetzung für erfolgreiche operationsfeldnahe periphere Blockaden. Wenn die Wirkungsweise der Lokalanästhetika, ihre toxischen Effekte und potenzielle Arzneimittelinteraktionen bei ihrem Metabolismus in der Praxis beachtet werden, dann ist das Risiko von Komplikationen relativ gering. Es sollte dennoch nicht unterschätzt, und adäquate Notfallmaßnahmen im Operationsteam sollten regelmäßig trainiert werden.     

Leprosy in a University Hospital in Southern Brazil

BACKGROUND

Leprosy is an infectious disease that may lead to irreversible nerve damage, compromising patient''s quality of life and leading to loss of working years.

OBJECTIVES

To evaluate the epidemiological profile of patients followed at a University Hospital.

MATERIALS AND METHODS

This is a retrospective observational study, based on a review of medical records. We studied the clinical and epidemiological features of patients with leprosy monitored at the Hospital de Clínicas of the Federal University of Paraná between January 2005 and January 2010.

RESULTS

The mean age was 47.51, while 35.94% of patients were aged 41-60. The male:female rate was 1.8:1. The most prevalent occupations were: retired, students or rural workers. Patients came mainly from Curitiba or nearby areas, but there were also patients from the countryside. The mean diagnostic delay was 24.57 months. Multibacillary forms prevailed, with the lepromatous variety being the most common, closely followed by the borderline type. Neural enlargement was found in more than 50% of the patients and 48.44% of them developed reactional states. Hemolysis was the most commonly detected drug side effect. Initial functional evaluation was possible in 70% of patients, 55% of whom had disabilities upon diagnosis. The most prevalent associated disease was hypertension.

CONCLUSIONS

This study showed an important diagnostic delay and a high rate of sequelae in this specific population. Brazil is one of the few remaining countries that has not yet eradicated leprosy and it is important to improve health policies in order to prevent sequelae and achieve eradication.     

Metástasis cutáneas de tumores sólidos. Estudio descriptivo retrospectivo

Introduction

Cutaneous metastases are a rare event compared with other metastases. Their incidence is estimated between 0.7% and 9% of patients with cancer. Their presence always indicates a disseminated malignant disease and could also be the first manifestation of a tumor, appearing simultaneously or after a diagnosis of cancer.

Objectives

To describe the epidemiological and clinical characteristics of cutaneous metastases in our hospital.

Material and method

We performed an observational retrospective study. We included all patients with cutaneous metastases diagnosed in the Dermatology Service of our hospital during a 7-year period. Patients with metastases from sarcomas, melanomas and hematologic malignances were excluded.

Results

We studied 102 patients with cutaneous metastases. The primary tumor was identified in 87 patients (85%). The most frequent tumors were breast cancer in women (48%) and lung cancer in men (11%). Cutaneous metastases were the first sign of malignancy in 12 patients (11%). The predominant clinical form was the nodular form. The most common localization of the metastases was the thoracic region. The most common histologic diagnosis was adenocarcinoma. The mean survival after the development of metastases was 22.1 months.

Conclusion

There is an association between the frequency of cutaneous metastases and the most frequent malignancies in each sex.Our results were similar to those of other studies regarding age, sex distribution, the predominant clinical form, location, and histological diagnosis of cutaneous metastases.