无毛小鼠接种申克孢子丝菌检测抗申克孢子丝菌细胞外蛋白酶抗体研究

通过申克孢子丝菌接种无毛小鼠（Ｈａｉｒｌｅｓｍｉｃｅ）来检测两种细胞外蛋白酶的血清抗体。在小鼠皮内注射申克孢子丝菌并形成结节，镜下示真菌消失，并在每周通过酶联免疫吸附试验（ＥＩＡ）检测蛋白酶Ⅰ和Ⅱ的血清抗体。结果显示抗体滴度与实验小鼠孢子丝菌病肉眼和镜下观察的变化趋势相一致。本试验证明了申克孢子丝菌在体内能产生蛋白酶Ⅰ和Ⅱ，而且蛋白酶的抗体可用于孢子丝菌病的血清学诊断。     

无毛小鼠接种申克孢子丝菌检测抗申克孢子丝菌细胞是白酶…

通过申克孢子丝菌接种无毛小鼠（Ｈａｉｒｌｅｓｓ ｍｉｃｅ）来检测两种细胞外蛋白酶的血清抗体。在小鼠皮内注射申克孢子丝菌并形成结节，镜下示真菌消失，并在每周通过酶联免疫吸附试验（ＥＩＡ）检测蛋白Ⅰ和Ⅱ的血清抗体。结果显示抗体滴度与实验小鼠孢子丝菌病肉眼和镜下观察的变化趋势相一致。本试验证明了申克孢子丝菌在体内能产生蛋白酶Ⅰ和Ⅱ，而且蛋白酶的抗体可用于孢子丝菌病的血清学诊断。     

申克孢子丝菌病3例报告

我科在1998年～2001年发现3例孢子丝菌病,现报告如下。1 病例报告 例1,患者,男,45岁,1998年7月就诊。病史6年,发病前是农民,有外伤史,现为个体户,广东籍。右踝部一鹤鹑蛋大紫红色呈疣状增生样结节,近半年来破溃、糜烂、与皮下粘连。近2个月     

阴囊孢子丝菌病1例及申克孢子丝菌基因型检测

孢子丝菌病是由申克孢子丝菌引发的一种皮肤、皮下组织和附近淋巴管的慢性感染性疾病,多发于四肢等容易暴露受伤的部位.本文报道1例发生于阴囊的固定型皮肤孢子丝菌病,并对申克孢子丝菌进行了形态学和分子生物学水平的鉴定.     

申克孢子丝菌的分子生物学研究进展

申克孢子丝菌是孢子丝菌病的致病菌，以双相致病真菌。发生孢子丝菌病是由于宿主体内酵母细胞增殖所致，同时还与宿主防御机制有关。对其基因分型、部分基因的鉴定、分子诊断方法及致病机理的分子基础等方面的研究进展进行了综述。     

申克孢子丝菌的分子生物学研究进展

申克孢子丝菌是孢子丝菌病的致病菌 ,为双相致病真菌。发生孢子丝菌病是由于宿主体内酵母细胞增殖所致 ,同时还与宿主防御机制有关。对其基因分型、部分基因的鉴定、分子诊断方法及致病机理的分子基础等方面的研究进展进行了综述     

申克孢子丝菌的体外耐热实验观察

申克孢子丝菌是孢子丝菌病的病原菌,该菌为双相性真菌,本实验通过不同的温度刺激体外培养的酵母型孢子丝菌,观察其形态结构的变化、生长繁殖规律及最低温度的致死率,进一步探讨孢子丝菌病温热疗法的作用机理,以对孢子丝菌病的治疗提供新的方法。一、资料和方法筛选1例固定型孢子丝菌病患者皮损处组织块及渗出液,用沙堡培养基27℃培养一周,将此菌落接种在脑心葡萄糖血琼脂培养基(BHI)上,36℃培养一周,以此菌株做为本次实验的实验菌株。     

申克孢子丝菌基因型与临床表现型关系研究

目的了解申克孢子丝菌的基因学特征，探索其基因型特征与临床表现型的关系。方法收集不同临床型别孢子丝菌病的申克孢子丝菌分离株并提取DNA，再用随机扩增DNA多态性方法（RAPD）进行PCR扩增。结果①共选用10个随机引物进行扩增，筛选出4个具有较好扩增片段的引物。②不同菌株同-引物扩增均见-共有DNA片段。③不同临床型孢子丝菌病的申克孢子丝菌分离株带型有差异。结论随机扩增DNA多态性研究发现申克孢子丝菌具有一定的种内差异，其DNA带型与菌株的临床型有关。     

皮肤淋巴管型孢子丝菌病1例

患者女,57岁。右拇指、腕部及前臂结节伴瘙痒和疼痛3月。右前臂皮损组织病理示:化脓性肉芽肿性改变。真菌培养见申克孢子丝菌生长。诊断:皮肤淋巴管型孢子丝菌病。     

申克孢子丝菌TPS基因生物信息学分析

目的:分析和预测申克孢子丝菌海藻糖磷酸合成酶(TPS)基因及其编码蛋白的生物信息学特征及功能。方法:利用生物信息学软件和网站分析预测申克孢子丝菌TPS基因及其编码蛋白的结构和功能特征。结果:申克孢子丝菌TPS基因全长为2 864 bp,编码905个氨基酸。Target P和MotifScan预测该蛋白定位于胞浆中,具有糖基转移酶、海藻糖磷酸酶两大功能域及丰富的修饰位点,其中磷酸化位点尤为丰富,包含cAMP依赖性磷酸激酶、蛋白激酶C(PKC)和酪氨酸激酶(CK2)磷酸化位点。TPS蛋白是亲水性蛋白,无跨膜结构,未发现信号肽。结论:预测申克孢子丝菌TPS基因编码蛋白位于胞浆中,具有糖基转移酶、海藻糖磷酸酶两大功能域及丰富的修饰位点的结构功能特点,可行相应小分子抑制剂筛选以开发新型抗真菌药物。     

小鼠实验性孢子丝菌病病理学研究——正常和免疫损害小鼠白细胞功能观察

通过小鼠模型研究机体对申克孢子丝菌的防御机理。正常小鼠在接种申克孢子丝菌后 ,伴随着 PMNs的浸润 ,真菌发生裂解并逐渐消失 ;而裸鼠接种菌体后 ,在早期防御过程中因 PMNs杀伤活性的缺乏导致真菌繁殖扩散 ,直至动物死亡。     

Effects of a sunscreen formulation on albino hairless mice: a morphological approach

The purpose of the study was to evaluate the effects of a sunscreen formulation on the skin of albino hairless mice subjected to simulated solar light (SSL) in terms of morphological changes. Young adult albino hairless mice HRS/J (n = 36) were used as an experimental model for determining skin photoaging changes. Mice were irradiated with SSL, and the sunscreen (estimated SPF 30, PF-UVA) was obtained from the Pharmacy College/UFRJ, Brazil. The animals were divided into four groups: non-treated (G1), radiation only (G2), sunscreen-treated (G3) and vehicle + radiation (G4). Animals from groups G2, G3 and G4 were irradiated weekly (5 weeks), with no immobilization. One week after the final exposure, the dorsal skin was observed using a dermatoscopic camera. Biopsies were analyzed in order to quantify neovascularization and to evaluate histological aspects of the skin. Neovascularization was also evaluated with immunohistochemical reactions for the Von Willebrand factor. Animals from G2 displayed classical morphological changes denoting skin photoaging: thickening of the epidermis, increased dermal cellularity, follicular keratosis, sebaceous gland hyperplasia, and angiogenesis. Animals from groups G3 and G1 displayed similar morphological profiles, without these changes. Animals from group G4 showed more morphological changes than group G2, emphasizing the relative importance of the putative photosensitizing components present in the vehicle formulation. The extent of the morphological skin changes suggested that the sunscreen formulation was effective against SSL, and showed the importance of assessing the phototoxicity of vehicle formulations.     

Effects of near-infrared radiation on the epidermal proliferation and cutaneous immune function in mice

While ultraviolet radiation alters various cutaneous cell functions, little is known about the photobiological effects of infrared radiation (IR) on the skin except its local thermal effect. This study demonstrated that single exposure of mouse skin to near IR (0.7-1.3 μm) reversibly suppressed the proliferating activity of the epidermis, the density of Langerhans cells, and the ability of skin to induce contact hypersensitivity reaction. During the exposure, the ear surface temperature was elevated from a mean of 27 to 31.2°C. The results suggest that near IR can modulate the epidermal proliferation and part of the skin immune system, with a mild thermal effect.     

Effects of all-trans retinoic acid on UVB-irradiated and non-irradiated hairless mouse skin.

The effects of all-trans retinoic acid (t-RA) on photodamaged and normal non-irradiated skin were examined in hairless mice (Skh:HR-1). After being exposed to increasing doses of UVB for 10 weeks (total dose = 1.4 J/cm2), the animals were then treated with 0.1% t-RA in ethanol (50 microliters, five times per week) for another 10 weeks. Several animals (the follow-up group) were further observed after the termination of the t-RA treatment to investigate if the t-RA effect was reversible. Wrinkle effacement induced by t-RA was compared with three other parameters: a) de novo collagen synthesis, b) width of the dermal repair zone, and c) epidermal thickening. Interestingly, t-RA did not stimulate collagen synthesis in animals not exposed to UVB. In the irradiated animals, the time course of wrinkle reduction correlated with the stimulation of collagen synthesis. After a synchronous initial lag phase of 4-6 weeks, the wrinkling decreased from the maximum grade of 4 to a mean grade of 1.3, whereas collagen synthesis was enhanced to 245% of the control at week 10 of t-RA treatment. In contrast, a similar lag phase was not observed for either the appearance of the dermal repair zone or epidermal thickening. In the follow-up group, upon termination of t-RA treatment, collagen synthesis returned to the control level. Wrinkle effacement and thickening of the dermal repair zone, however, did not regress, suggesting the anti-photoaging effect of t-RA was not reversible over this time frame. The correlation between the length of the lag phases for collagen synthesis and wrinkle reduction points to the possibility that collagen plays an important role in tRA-induced wrinkle effacement. Both parameters are thus important endpoints for investigating the mechanism of RA-induced repair of photodamaged skin.     

The effects of topical alpha-hydroxyacids on the normal skin barrier of hairless mice

BACKGROUND: alpha-hydroxyacids (AHA), such as glycolic acid and lactic acid, have recently been used in cosmetic and dermatological formulations. However, the mechanisms of action of these substances have not been well documented. OBJECTIVES: This study was done to investigate the effects of AHA on the skin barrier of hairless mice and to clarify the contribution of AHA to the formation and secretion of the lamellar bodies (LB), which are known to be the critical structure for barrier function in the epidermis. METHODS: 5% Lactic acid and 5% glycolic acid were applied to normal skin of the mice daily for 14 days. RESULTS: Changes in transepidermal water loss (TEWL), capacitance and electron microscopic findings of the epidermis of hairless mice were compared with those in which only the vehicle was applied. CONCLUSIONS: There were no significant differences in TEWL, capacitance or epidermal thickness between the epidermis of the mice to which AHA or vehicle only had been applied. On electron micrographs, the normal epidermis to which AHA had been applied showed an increase in the number and secretion of LB and a decrease in the number of stratum corneum (SC) layers in comparison with the epidermis to which the vehicle only had been applied. The lipid layers of the SC intercellular spaces and calcium gradient in both the epidermis with application of AHA and that with vehicle only were normal. These results suggest that AHA, in low concentration (5%), may improve the skin barrier in hairless mice by inducing enhanced desquamation, and by increasing the number and secretion of LB without increasing TEWL.     

Effects of indomethacin and dexamethasone on mechanical scratching-induced cutaneous barrier disruption in mice

Effects of indomethacin and dexamethasone on recovery of cutaneous barrier disruption induced by mechanical scratching were examined. Cutaneous barrier was disrupted by scratching using a stainless-steel wire brush (mechanical scratching) and compared to cutaneous application of acetone/ether (1:1) mixture (AE) and tape-stripping. Increase of transepidermal water loss (TEWL), as an indicator of a broken skin barrier, and recovery period for mechanical scratching were higher and longer than those for AE treatment and tape-stripping and we also confirmed the severity of skin damage in a histological study. Topical application of moisturizers showed a temporal effect, rapidly decreased TEWL on mechanical scratching- or AE treatment-induced cutaneous barrier disruption, and gradually increased base levels from 4 to 12 h after treatment. Topical application of indomethacin or dexamethasone prolonged the recovery period for the cutaneous barrier, and concomitant use further worsened the status of the barrier. Additionally, we examined the effects of prostaglandins (PGs) and inflammatory cytokine on mechanical scratching-induced cutaneous barrier disruption pretreated with indomethacin and dexamethasone. As a results, PGD2 and interleukin (IL)-1beta significantly accelerated the recovery of cutaneous barrier disruption by mechanical scratching but such was not the case with PGE2, IL-1alpha, and tumor necrosis factor-alpha treatment. These results suggest that indomethacin and dexamethasone prolonged the recovery period caused by inhibition of PGD2 and IL-1beta. Mechanical scratching-induced cutaneous barrier disruption may be a useful method for evaluating means of recovery from skin damage.     

扇贝多肽对UVB损伤无毛小鼠皮肤结构及其抗氧化剂含量的影响

目的 :　本文探讨扇贝多肽 (PCF)对中波紫外线 (UVB)照射下小鼠皮肤的氧化损伤有无保护作用。方法 :　建立UVB(辐照强度为 5 .15× 10 - 2 J cm2 × 30天 )对无毛小鼠皮肤氧化损伤模型。昆明种无毛小鼠 ,随机分为双蒸水未照射组和UVB模型组 (双蒸水对照组、5 %PCF组、2 0 %PCF组、10 %维生素C组 )。光镜下测表皮平均厚度及成纤维细胞数目 ;电镜观察皮肤组织超微结构。酶法测定皮肤匀浆上清液中抗氧化酶 (谷胱甘肽GSH -Px、超氧化物歧化酶SOD、)的活性和丙二醛 (MDA)的含量及总抗氧化能力(T -AOC)。结果 :　①UVB损伤的对照组小鼠皮肤光镜下可见表皮变薄、成纤维细胞数量减少 ;电镜下表皮细胞胞质内可见空泡形成 ,真皮的成纤维细胞内可见囊泡状扩张的滑面内质网、粗面内质网等细胞器减少。②PCF能增加表皮的平均厚度和皮肤成纤维细胞的数量 ;超微结构显示PCF组表皮细胞结构正常 ,成纤维细胞的粗面内质网明显增多 ,与维生素C的抗氧化作用一致。③PCF可提高UVB辐射损伤小鼠皮肤组织的总抗氧化能力、SOD活性 ,降低MDA含量 ;与维生素C的抗氧化作用一致。结论 :　扇贝多肽与VitC一样具有抗UVB氧化损伤的作用。其作用机制可能与扇贝多肽提高抗氧化酶含量、清除自由基有关     

Influence of culturing temperature and proteinase inhibitors on the spontaneously occurring changes in the organ culture of psoriatic skin.

Skin explants from involved psoriatic lesions showed dissociation of keratinocytes, dermal-epidermal separation and degenerative changes such as cytoplasmic swelling of subcorneal prickled cells within 24 h after culture initiation at 37 degrees C in the absence of fetal bovine serum (FBS). These histological changes developed time dependently, while normal skin explants did not exhibit such phenomena. Some of the uninvolved psoriatic skin explants showed only degenerative change 48 to 72 h after culture initiation at 37 degrees C. To determine the nature of these spontaneously occurring changes in psoriatic skin explants and then to approach the pathogenesis of psoriasis, the effects of FBS, various proteinase inhibitors (PIs) and culturing temperature (37, 31, 24 degrees C) were examined in skin organ culture of normal and involved psoriatic skin. At 37 degrees C, only serine PIs (5 or 10 mg/ml of soybean trypsin inhibitor (SBTI), 1000 KIU/ml of aprotinin, or 2 mg/ml of camostat mesilate in the medium) or FBS (20% in the medium) could suppress the occurrence of dissociation of keratinocytes and dermal-epidermal separation but not the degenerative change in involved psoriatic skin explants, while other types of PIs did not exhibit any such inhibition. When the culturing temperature was reduced from 37 degrees C to 31 or 24 degrees C, the formation of dissociation of keratinocytes and dermal-epidermal separation was almost non-existent and only moderate degenerative change was observed. The addition of FBS or serine PIs to the culture at 31 degrees C revealed the formation of very weak degenerative change.(ABSTRACT TRUNCATED AT 250 WORDS)     

Effect of infrared radiation A on photoaged hairless mice harboring eumelanin and pheomelanin in the epidermis

Infrared radiation A (IRA) is absorbed by melanin and generates heat. Therefore, the effect of IRA could be well analyzed using skin, which contains melanin in the epidermis. Hairless mice harboring epidermal melanocytes that produce eumelanin, pheomelanin, or non‐melanin were generated by backcrossing K14‐stem cell factor mice, recessive yellow mice, and then albino hairless mice. High‐dose IRA was irradiated over 18 weeks after the establishment of photoaged mice by irradiation with ultraviolet B (UVB) three times a week for 14 weeks. Tumor formation was assessed every week. The formation of cyclobutane pyrimidine dimer and apoptotic cells by the irradiation of IRA and UVB was evaluated. Repetitive irradiation of IRA did not promote tumor formation in all types of mice. Pre‐irradiation of IRA to UVB, but not post‐irradiation, accelerated the elimination of cyclobutane pyrimidine dimers and enhanced apoptosis; these effects were most obvious in eumelanin‐producing mice. Real‐time polymerase chain reaction analysis showed downregulation of FLICE (cellular caspase 8)‐like inhibitory protein and B‐cell lymphoma‐extra large and upregulation of Bcl‐2‐associated X protein by UVB, but further enhancement of these molecules by pre‐irradiation of IRA was not observed. These results indicate that IRA does not confer the promotion of UVB‐induced carcinogenesis in photoaged mice harboring epidermal melanocytes and that photochemical reaction between IRA and melanin might be involved in the induction of apoptosis and the elimination of cyclobutane pyrimidine dimers by UVB. The enhancement of apoptosis by pre‐irradiation of IRA to UVB might be induced by mechanisms other than the modification of the mRNA expression of FLICE (cellular caspase 8)‐like inhibitory protein, B‐cell lymphoma‐extra large, and Bcl‐2‐associated X.