白色念珠菌分泌型天冬氨酸蛋白酶在口腔扁平苔藓患者唾液中的表达

研究白色念珠菌分泌型天冬氨酸蛋白酶(SAP)家族在扁平苔藓患者唾液中的表达,进而探讨白色念珠菌病与扁平苔藓的关系。选取白色念珠菌携带者、白色念珠菌病患者、携带白色念珠菌的扁平苔藓患者、伴有白色念珠菌病的扁平苔藓患者各20例,念珠菌培养阴性的扁平苔藓患者10例。从唾液标本中直接提取RNA,进行RT-PCR。白色念珠菌携带者唾液中有SAP2、SAP4-6、SAP7的不同表达,携带白色念珠菌的口腔扁平苔藓患者唾液中有SAP2、SAP4-6、SAP7、SAP8、SAP9的不同表达,所有携带者中均未表现SAP1、SAP3和SAP10的阳性扩增。白色念珠菌患病者及伴有白色念珠菌病的口腔扁平苔藓患者唾液中有SAP1-10的不同表达。念珠菌培养阴性的扁平苔藓患者均无SAP1-10的阳性扩增。SAP9与扁平苔藓密切相关。     

三联疗法治疗糜烂型扁平苔藓的疗效观察

目的:探讨肿痛安联合曲安奈德、高压氧治疗糜烂型扁平苔藓(OLP)的临床疗效.方法:69例糜烂型口腔扁平苔藓患者分成肿痛安联合曲安奈德治疗组和对照曲安奈德两组;在治疗组采用肿痛安口服加曲安奈德注射于粘膜下层治疗糜烂型OLP,同时配合高压氧治疗.而对照组采用曲安奈德局部注射治疗.结果:治疗组中有效例数为34例,总有效率为92.1%;对照组中有效例数23例,总有效率为74.1%,治疗组与对照组比较,差异具有显著性(p<0.05).结论:采用肿痛安口服联合曲安奈德注射于粘膜下层治疗糜烂型OLP配合高压氧的"三联疗法"疗效确切,副作用小,值得推广.     

家蝇抗菌肽AMP-17对白色念珠菌菌丝的抑制作用

【背景】AMP-17是从微生物诱导的家蝇转录组数据库筛选到的一条特异性高表达基因,采用原核表达体系获得其重组蛋白并证实了具有显著的抗菌效果,特别是对白色念珠菌具有较强的抗菌活性。【目的】研究抗菌肽AMP-17对白色念珠菌菌丝的抑制作用。【方法】采用微量液体稀释法测定AMP-17对11株白色念珠菌的最小抑菌浓度(minimal inhibitory concentration,MIC);根据对AMP-17的敏感程度选取3株绘制生长曲线;通过光学显微镜观察并计数经AMP-17作用后白色念珠菌芽生孢子生成率及芽管形成率;倒置荧光显微镜观察白色念珠菌酵母相向菌丝相转化及以菌丝相为起点AMP-17促进菌丝相转化为酵母相的情况。【结果】AMP-17对支气管肺泡灌洗液分离株16105的MIC为10μg/mL,对粪便分离株16214的MIC为40μg/mL,对其余9株白色念珠菌的MIC均为20μg/mL;白色念珠菌经不同浓度的AMP-17作用后,各时间点的芽生孢子生成率均显著低于对照组,尤其是40μg/mL的AMP-17组,芽生孢子生成率仅15%,显著低于阳性药物氟康唑;各实验组芽管形成率显著低于对照...     

白芍总苷对糜烂型口腔扁平苔藓外周血TNF-α，IFN-γ及IL-10水平的影响

目的:研究白芍总苷对糜烂型口腔扁平苔藓外周血肿瘤坏死因子(TNF-α)、γ-干扰素(IFN-γ)及白细胞介素-10(IL-10)水平的影响。方法:选取2014年7月至2015年7月我院接诊的糜烂型口腔扁平苔藓患者80例作为本次研究对象。对照组患者采用低浓度他克莫司治疗,观察组采用白芍总苷治疗,观察两组患者治疗前后血清TNF-α、IFN-γ及IL-10水平的变化、糜烂面积、疼痛评分及近期治疗疗效。结果:治疗后,观察组血清TNF-α、IL-10水平显著低于对照组,血清IFN-γ水平明显高于对照组[(2.16±0.61)μg/m L vs(3.04±0.80)μg/m L、(258.93±5.72)ng/L vs(273.41±6.03)ng/L、(319.27±53.46)ng/L vs(290.95±51.03)ng/L(P0.05),糜烂面积、疼痛评分均显著低于对照组[(0.10±0.03)cm2 vs(0.51±0.10)cm~2、(1.01±0.30)分vs(3.20±0.78)分](P0.05),近期治疗疗效优于对照组95.00%(38/40)vs75.00%(30/40)(P0.05)。结论:白芍总苷治疗糜烂型口腔扁平苔藓的效果显著,可能与其调节血清TNF-α、IFN-γ及IL-10的水平有关。     

口腔扁平苔藓者白色念珠菌检出率及分离株的磷脂酶活性研究

目的:研究健康人和口腔扁平苔藓(oral lichen planus,OLP)患者的口腔白色念珠菌检出率及分离株的磷脂酶活性.方法:采用含漱液浓缩培养法及白色念珠菌微生物学鉴定法,考察健康对照组、糜烂型和非糜烂型OLP患者的白色念珠菌检出率;应用蛋黄培养基沉淀环面积测量法,比较来自于健康人(26株)、糜烂型OLP患者(62株)以及非糜烂型OLP患者(24株)共112株白色念珠菌的磷脂酶活性大小.结果:糜烂型OLP患者组的口腔白色念珠菌检出率高于健康对照组和非糜烂型扁平苔藓患者组;糜烂型和非糜烂型OLP患者组的白色念珠菌分离株的磷脂酶活性高于健康对照组.结论:两种类型的OLP患者组的白色念珠菌分离株与健康对照组相比,具有不同的毒性特征,白色念珠菌与OLP(尤其是糜烂型)发生发展可能相关.     

硫酸羟氯喹口服辅助治疗糜烂型口腔扁平苔癣的效果及对AT-Ⅲ、ZAG的影响

摘要 目的：探讨硫酸羟氯喹口服辅助治疗糜烂型口腔扁平苔癣的效果及对人抗凝血酶Ⅲ（AT-Ⅲ）、锌α2糖蛋白（ZAG）的影响。方法：选择2018年12月-2021年1月在我院接受治疗的115例糜烂型口腔扁平苔癣患者,采用随机数表法分为治疗组（n=58）和对照组（n=57）。对照组给复方倍他米松治疗,治疗组在对照组的基础上联合硫酸羟氯喹口服治疗。比较两组临床疗效、AT-Ⅲ、ZAG、白细胞介素4（IL-4）、白细胞介素17（IL-17）、干扰素γ（IFN-γ）、糜烂面积、疼痛评分水平变化情况及不良反应发生情况。结果：治疗后,两组总有效率比较差异显著（P<0.05）;治疗前,治疗组和对照组血清AT-Ⅲ、ZAG比较无显著差异;治疗后,治疗组和对照组血清AT-Ⅲ、ZAG均随着时间的推移而降低,且治疗组均低于对照组,差异显著（P<0.05）;治疗前,治疗组和对照组实验室指标水平比较无显著差异;治疗后,治疗组和对照组IL-4、IL-17均随着时间的推移而降低,且治疗组均低于对照组,IFN-γ均随着时间的推移而升高,且治疗组均高于对照组,差异显著（P<0.05）;治疗前,治疗组和对照组糜烂面积、疼痛评分比较无显著差异;治疗后,治疗组和对照组糜烂面积、疼痛评分均随着时间的推移而降低,且治疗组均低于对照组,差异显著（P<0.05）;两组不良反应总发生率为3.45%、8.77%,无显著差异（P>0.05）。结论：在糜烂型口腔扁平苔癣中应用硫酸羟氯喹口服辅助治疗疗效显著,可有效改善患者AT-Ⅲ、ZAG水平。     

Th1型和Th2型细胞因子在口腔扁平苔藓患者外周血中的表达及临床意义

目的:研究口腔扁平苔藓(OLP1)患者Th1型和Th2型细胞因子的表达及临床意义。方法:选取2013年1月至2014年3月于我院就诊的21例充血糜烂型及14例光滑型OLP患者为研究对象,18例正常人为对照组,采用密度梯度离心法对各组外周血单个核细胞进行分离,酶联免疫吸附剂测定(ELISA)法对各组外周血单个核细胞中的IL-4和IFN-gamma的表达进行检测,逆转录-聚合酶链反应法对各组血清中IL-4 m RNA和IFN-gamma m RNA的表达进行检测。结果:与正常对照组相比,OLP患者IL-4m RNA及蛋白的表达均增高,而IFN-gamma m RNA及蛋白的表达则降低,差异均有显著统计学意义(均P0.01)。充血糜烂型及光滑型OLP患者组间比较发现,IL-4 m RNA和IFN-gamma m RNA及蛋白的表达差异无统计学意义(P0.05)。结论:OLP发病机制与Th1与Th2的表达失衡有关,为临床治疗提供参考。     

饲料中添加抗菌肽Surfactin对吉富罗非鱼肠道健康的影响

为研究饲料中添加抗菌肽Surfactin对吉富罗非鱼(GIFT, Oreochromis niloticus)肠道健康指标的影响, 实验将平均体重为(12.010.03) g/尾的320尾吉富罗非鱼, 随机分为4组, 每组4个重复, 每个重复20尾鱼, 分别投喂抗菌肽Surfactin添加水平为0(对照组)、50、100和200 mg/kg的试验饲料7周。结果显示: 与对照组相比, 吉富罗非鱼饲料中添加抗菌肽Surfactin可使肠道皱襞高度显著增加(P0.05), 肌层厚度无显著变化(P0.05); 大肠杆菌数量显著降低(P0.05), 乳酸杆菌数量显著增加(P0.05), 细菌总数无显著变化(P0.05); 显著提高肠道总抗氧化能力水平、谷胱甘肽过氧化物酶活性和超氧化物歧化酶活性(200 mg/kg抗菌肽添加组除外)(P0.05), 显著降低肠道丙二醛水平(P0.05), 对过氧化氢酶活性无显著影响(P0.05); 吉富罗非鱼肠道健康指标以50 mg/kg抗菌肽Surfactin添加组最佳。试验表明, 吉富罗非鱼饲料中适量添加抗菌肽Surfactin可增加肠道皱襞高度、调节肠道菌群和提高肠道抗氧化能力而改善肠道健康状态。     

改良型抗菌肽的研究进展

长期滥用抗生素导致了耐药菌株“超级细菌”的出现,增加了动物、人类健康和环境污染风险．寻找抗生素替代品正成为全球研究热点,抗菌肽因其高效抗菌效果和不同于抗生素的独特作用机制引起了各国研究者的关注,并进行了相关研究．然而抗菌肽的安全性、稳定性、生产成本等问题限制了其生产与应用．为了克服这些不利因素,研究者们对抗菌肽进行了多种方式的改造,产生了模拟型、同源型、杂合型、轭合型、稳定型和固位型等改良型抗菌肽,并有望在畜牧业、食品业、医药业等领域得到广泛的应用．本文主要综述了这些改良型抗菌肽近年来的研究进展．     

抗菌肽Cecropin B对水稻植株生长的影响

ＣｅｃｒｏｐｉｎＢ是抗菌肽家族中的一员,具有较广的杀菌谱,因此被应用于植物基因工程领域提高植物的抗病性。我们取转ＣｅｃｒｏｐｉｎＢ基因水稻（Ｏｒｙｚａｓａｔｉｖａ）品种京引１１９（ＪＹ１１９）及其后代不同株系,于１９９７年夏,直播于本所田间同室隔离区,每株系１００株。分蘖末期接种白叶枯病菌９６１２（浙江农科院植保所提供）,接种浓度约１０９ＣＰＵ,每株用手术剪接种４～５叶。１０ｄ后测定每株白叶枯病菌侵染长度。随后用农药叶青双可湿性粉剂０．２％（Ｗ／Ｖ）喷施,将白叶枯病菌杀死。待植株生长恢复后,于齐…     

Onychomycosis Incidence in Type 2 Diabetes Mellitus Patients

The onychomycosis incidence was determined in 250 type 2 diabetes mellitus (T2DM) patients who were registered at the Internal Medicine Service from a Mexico city General Hospital throughout a year (January-December 2006). Out of the total of studied T2DM patients, 93 (37.2%) showed ungual dystrophy and from these, in 75.3% a fungal etiology was corroborated. Out of 70 patients, 34 were men and 36 women, with an average of 63.5 years. Correlation between T2DM evolution time and onychomycosis was significant (P < 0.01). Distal-lateral subungual and total dystrophic onychomycosis were the most frequent clinical types (55.1% and 33.7%, respectively). Fifty-eight fungal isolates were obtained; 48.6% corresponded to dermatophytes, Trichophyton rubrum being the first species (37.1%). All these strains corresponded to two morphological varieties: "yellow" and typical downy. From the yeast-like isolates, 12 corresponded to Candida spp., firstly C. albicans and C. parapsilosis; three to Cryptococcus spp. (C. albidus, C. uniguttulatus and C. laurentii); two Trichosporon asahii; and only one to Pichia ohmeri. Six non-dermatophytic molds were isolated: two Chrysosporium keratinophylus, two Scopulariopsis brevicaulis, one Aspergillus fumigatus, and one Acremonium sp. The fungal mixture corresponded to T. mentagrophytes with C. guilliermondii; T. mentagrophytes with C. glabrata; T. rubrum with C. glabrata; T. rubrum with P. ohmeri.     

The white-phase-specific gene<Emphasis Type="Italic"> WH11</Emphasis> is not required for white-opaque switching in<Emphasis Type="Italic"> Candida albicans</Emphasis>

Phenotypic switching between white and opaque cells is important for adaptation to different host environments and for mating in the opportunistic fungal pathogen Candida albicans. Genes that are specifically activated in one of the two cell types are likely to be important for their phenotypic characteristics. The WH11 gene is a white-phase-specific gene that has been suggested to be involved in the maintenance of the white-phase phenotype. To elucidate the role of WH11 in white-opaque switching, we constructed mutants of the C. albicans strain WO-1 in which the WH11 gene was deleted. The wh11 mutants were still able to form both white and opaque cells whose cellular and colony phenotypes were indistinguishable from those of the wild type. Deletion of WH11 also did not affect the activation and deactivation of the white-phase-specific WH11 promoter and the opaque-phase-specific OP4 and SAP1 promoters in the appropriate cell type. Finally, switching from the white to the opaque phase and vice versa occurred with the same frequency in wild-type and wh11 mutants. Therefore, the WH11 gene is not required for phenotypic switching, and its protein product seems to have other roles in white cells, which are dispensable after the switch to the opaque phase.Communicated by E. Cerdá-Olmedo     

抗植物病原真菌紫叶小檗内生真菌的筛选和鉴定

对紫叶小檗根、茎、叶和果实中分离纯化得到的28株内生真菌进行液体培养,培养物烘干研磨后用丙酮浸提,测定各提取物对5种供试植物病原真菌的抑菌活性,对抗菌谱较广的菌株进行分子鉴定,并分别测定其胞内、胞外代谢产物的抑菌活性.结果显示:(1)紫叶小檗25株内生真菌中有5株菌(R3、S4、L6、F2、F6)的抗菌谱较广,其中,茎中的内生菌S4抑菌活性最强,对5种植物病原真菌的抑菌活性均大于90％,对马铃薯干腐病菌的抑菌率最高达93.06％.(2)经鉴定S4为子囊菌门的Para phaeos phaeria属真菌.(3)S4菌丝体丙酮提取液对马铃薯干腐病菌的抑菌率随其浓度(加入体积)的增加而增大,呈正相关线性关系(y=15.334x+14.618,r=0.99),而S4发酵液对马铃薯干腐病菌的抑菌率随其浓度(加入体积)的增加而减小,呈负相关线性关系(y=-20.196x+64.984,r=0.99),即在较高浓度时促进病原菌生长.研究表明,紫叶小檗内生真菌S4菌株的抑菌活性成分主要为胞内代谢产物.     

Phospholipase and Proteinase Activities of Clinical Isolates of Candida from Immunocompromised Patients

Sixty-one isolates of Candida recovered from HIV seropositive and cancer patients were studied for elaboration of putative virulence determinants – phospholipase (PL) and secreted aspartyl proteinase (Sap). Forty two (68.85%) isolates examined were PL producers and 51 (83.6%) were positive for Sap. 57.37% (35/61) isolates produced both enzymes. Enzymatic activity was more pronounced in Candida albicans with 100% PL and 94.1% Sap activity. In contrast, non-C. albicans species demonstrated only 29.6% PL and 70.3% Sap activity, indicating interplay of other virulence determinants in these yeasts in colonization and disease.     

New species and a new combination in the <Emphasis Type="Italic">Hyphopichia</Emphasis> and <Emphasis Type="Italic">Yarrowia</Emphasis> yeast clades

Three new species of Candida and a new combination in the genus Hyphopichia are proposed from phylogenetic analysis of nucleotide divergence in domains D1/D2 of the large subunit (26S) rDNA. The new taxa and their type strains are the following: Candida bentonensis sp. nov. (NRRL YB-2364, CBS 9994), Candida hispaniensis sp. nov. (NRRL Y-5580, CBS 9996), Candida pseudorhagii sp. nov. (NRRL YB-2076, CBS 9998) and Hyphopichia heimii comb. nov. (NRRL Y-7502, CBS 6139), basionym Pichia heimii Pignal. Phylogenetic analysis placed C. pseudorhagii and H. heimii in the Hyphopichia clade whereas C. bentonensis and C. hispaniensis are members of the Yarrowia clade.     

天蚕素A-马盖宁杂合肽对耐甲氧西林金黄色葡萄球菌生理代谢影响的研究

研究天蚕素A-马盖宁杂合肽作用耐甲氧西林金黄色葡萄球菌(MRSA)后对细胞生理代谢的影响。利用全波长酶标仪测定杂合肽作用MRSA后对DNA、RNA、总蛋白、β-半乳糖苷酶、碱性磷酸酶的影响,利用溶氧电极测定杂合肽对细胞呼吸作用的影响,通过生物发光分析仪检测杂合肽作用细胞后ATP生产的变化。结果显示,杂合肽作用MRSA后对细胞DNA、RNA、总蛋白的合成能力均出现抑制作用。β-半乳糖苷酶、碱性磷酸酶的表达活性同样受到了明显的抑制。杂合肽作用细胞后呼吸作用明显下降,ATP生产能力受到抑制。杂合肽作用细胞后抑制了胞内部分生物大分子的合成能力、胞内酶的表达活性及细胞的能量代谢功能,通过胞内机制发挥了抑菌作用。     

Comparison of the Influence of Carbon Substrates on the Fibrolytic Activities of Anaerobic Rumen Fungi

Cellulolytic and xylanolytic activities among genera of anaerobic fungi when grown on glucose, xylan and the cellulosic substrates, filter paper and Avicel were compared. All the fungi had basal extracellular fibrolytic activities that could be enhanced by growth on xylan or the cellulosic substrates. However,Piromyces communisstrain 22 andNeocallimastix patriciarumstrain 27 had substantially greater levels of fibrolytic activity thanOrpinomyces joyoniistrain 19-2 orNeocallimastix frontalisstrain RE1. Zymogram analysis suggested both structural and regulatory differences amongst the enzyme systems of the fungi. Numerous and varied enzyme bands were evidenced for all the fungi, with substantial substrate influences seen in the xylanase activities. Most commonly the smaller molecular weight bands, found exclusively extracellularly, appeared under the greatest regulatory control. Endoglucanase activities ofP. communisandO. joyoniidemonstrated similar regulatory control, while those of the twoNeocallimastixstrains did not appear to exert such control. These results suggest that while the enzymatic activities are functionally similar, there are likely significant variations in the enzyme systems of the anaerobic fungi.     

Pseudomonas aeruginosa, Candida albicans, and device-related nosocomial infections: implications, trends, and potential approaches for control

For many years, device-associated infections and particularly device-associated nosocomial infections have been of considerable concern. Recently, this concern was heightened as a result of increased antibiotic resistance among the common causal agents of nosocomial infections, the appearance of new strains which are intrinsically resistant to the antibiotics of choice, and the emerging understanding of the role biofilms may play in device-associated infections and the development of increased antibiotic resistance. Pseudomonas aeruginosa and Candida albicans are consistently identified as some of the more important agents of nosocomial infections. In light of the recent information regarding device-associated nosocomial infections, understanding the nature of P. aeruginosa and C. albicans infections is increasingly important. These two microorganisms demonstrate: (1) an ability to form biofilms on the majority of devices employed currently, (2) increased resistance/tolerance to antibiotics when associated with biofilms, (3) documented infections noted for virtually all indwelling devices, (4) opportunistic pathogenicity, and (5) persistence in the hospital environment. To these five demonstrated characteristics, two additional areas of interest are emerging: (a) the as yet unclear relationship of these two microorganisms to those species of highly resistant Pseudomonas spp and Candida spp that are of increasing concern with device-related infections, and (b) the recent research showing the dynamic interaction of P. aeruginosa and C. albicans in patients with cystic fibrosis. An understanding of these two opportunistic pathogens in the context of their ecosystems/biofilms also has significant potential for the development of novel and effective approaches for the control and treatment of device-associated infections.     

串联抗菌肽Cecropin B基因在莱茵衣藻中的表达及其抗菌活性分析

为了应对各种抗生素在水产养殖业所带来的副作用,我们在本文中尝试利用微藻对一种抗菌肽进行表达的可行性研究．根据莱茵衣藻核基因组偏爱密码子对抗菌肽Cecropin B基因进行改造,并将4个经改造的Cecropin B基因依次串联起来,中间加上莱茵衣藻的自剪切连接肽序列LWMRFA,人工合成总长度为522 bp的串联Cecropin B基因．将串联Cecropin B基因克隆到含hsp70-RBCS2启动子和RBCS2终止子的pH105载体上,再与携带ble筛选基因的表达框架连接,构建重组表达载体pCB124．采用玻璃珠转化法将载体pCB124导入莱茵衣藻cc-849中,筛选得到能表达抗菌肽Cecropin B的转基因衣藻．经过6个月的保持培养后,进一步对转基因藻细胞提取液进行抗菌活性分析,发现转基因藻具有明显的抑制革兰氏阴性菌(大肠杆菌)和革兰氏阳性菌(枯草芽孢杆菌和溶壁微球菌)生长的特征．这一结果为具有抗菌活性的饵料藻的生产和应用提供了新的途径．