马拉

篇一：6种中草药对马拉色菌的影响

学科分类号（二级）18021

本科学生毕业论文（设计）

题目 6种中草药抗马拉色菌的体外药敏试验

姓 名 李繁梅

学 号 094120086

院、 系 生命科学学院

专 业 生物科学

指导教师 张 田

职称（学历）副教授（硕士研究生）

6种中草药抗马拉色菌的体外药敏试验

出生日期：1990年6月12日

出生地：云南省曲靖市会泽县

QQ：269382864

电子邮箱：269382864@qq.com

手机号码：15198771591

专业：生物科学

摘要..........................................2

关键词........................................2

材料、设备及仪器..............................2

方法..........................................3

结果..........................................4

讨论..........................................5

参考文献......................................6

英文题目、摘要及关键词........................7

摘要：马拉色菌是一种条件致病性真菌，它会引起花斑藓、马拉色菌毛囊炎、银屑病、脂溢性皮炎等。目的：是筛选有效抑制马拉色菌的中草药。方法：采用微量稀释法与贴片法进行体外药敏实验。 结果：6种中草药（侧柏叶、三颗针、黄芩、丁香、何首乌、胡黄连）中侧柏叶和三颗针对马拉色菌有较强的抑制作用；黄芩和丁香对马拉色菌有一定的抑制作用；何首乌和胡黄连对马拉色菌没有抑制作用。

关键词：中草药； 马拉色菌； 抑制作用； 抑菌圈

马拉色菌（Malassezia)是人类及温血动物的正常菌群。是一种条件性致病性真菌，常驻皮肤的表面，具有酵母态和菌丝态。研究表明，该菌属与花斑糠诊、马拉色菌毛囊炎、脂溢性皮炎、特应性皮炎及一些银屑病的发生、发展密切相关

[1]。到二十世纪中期，人们基于形态学、超微结构、生理生化特征以及分子生物学的研究，将马拉色菌分为厚皮马拉色菌（M.Pachydermatis)、合轴马拉色菌(M.sympodialis)、糠秕马拉色菌(M.furfur)、钝性马拉色菌(M.obtusa)、球形马拉色菌(M.globosa)、限制性马拉色菌(M.restricta)、斯洛菲马拉色菌

(M.slooffiae)等七种[2-3]。近年来，Sugita等利用巢式PCR及PCR限制性片段长度多态（RFLP）等又发现3个新菌种。

随着现在临床上所用的纯西药抗头皮屑药物如氟康唑、酮康唑等对此虽有一定的效果，但从高效低毒以及耐药性等方面的评价，没有一种药物能够令人满意。抗真菌中药的筛选研究已开展多年，相比传统的唑类药物具有不良反应小、来源广、价格低廉、较小出现耐药等优点。开展针对马拉色菌的体外药敏实验筛选中 药，能够指导临床医生合理用药已受到越来越多的关注。

1材料、设备及仪器

1.1菌株

马拉色菌预计（35株），取自正常人

1.2培养基

橄榄油培养基：蛋白胨10g、葡萄糖40ml、酵母浸膏0.1g、单硬脂酸甘油酯2.5g、吐温-80 （重庆川江化学试剂厂；PH（50g/L,25℃):5.0-7.0)2.0ml、橄榄油（olive oil（折光率：1.468-1.471；皂化值：175-195；酸价：1-3；碘值：79-88）以不饱和脂肪酸为主;中国科学院昆明植物研究所)40g、琼脂18g、水1L[8]。

1.3药品：

侧柏叶、三颗针、丁香、黄芩、何首乌、胡黄连。

1.4阳性对照药品

酮康唑（口服抗真菌药，片装，每片含酮康唑0.2g；生产厂家：西安杨森制药有限公司；产品批号：090115144；批准文号：国药准字H10930212.），储备液用二甲基亚砜溶液配制。基于以往研究结果，配制酮康唑储备液质量浓度为320mg/l，氟康唑储备液质量浓度为1280mg/l。使用以前用无菌水稀释10倍。

1.5设备及仪器

1.5.1设备

电子天平，超净工作台，恒温烘箱，显微镜，手提式压力蒸汽灭菌锅(生产厂家：浙江新丰医疗器械有限公司），超声震荡机(Ultrasonic Cleaner,型号：DAS300-GL2-16L)。

1.5.2仪器

锥形瓶，小烧杯，玻璃棒，注射器，小勺，培养皿，移液枪，载玻片，盖玻片，滴管，量筒，小药瓶（容量：5ml和10ml)，微孔过滤器，打孔器。

2.方法

2.1配制培养基（100ml）

具体步骤如下：

在每次配制培养基之前都要对培养皿进行灭菌处理；

2.1.1称量:

蛋白胨：1g ； 葡萄糖：4g ； 酵母菌膏：0.01g ；单硬脂酸甘油脂：0.25g； 吐温80：0.2ml ； 橄榄油：4ml ； 氯霉素：0.005g 蒸馏水：100ml

2.1.2定容到100ml

2.1.3 灭菌：高压121℃,40min灭菌。

2.1.4倒平板:培养皿规格：100mm；高度：20mm；生长面积：55cm2 。

2.2对头皮屑进行镜检

2.2.1 配制体积分数为10%的KOH溶液和50%的双翎墨水（昆明市墨水厂出品）溶液

2.2.2 头皮屑经体积分数为10%KOH溶液处理，再用体积分数为50%的翎墨墨水溶液染色后在光镜下观察。

2.3菌种的培养

2.3.1 将头皮屑拨至预先消毒的纸片上，淋洒少量体积分数为75%的酒精于头皮屑上，待酒精挥发干后用无菌接菌针拨落头皮屑接种到含氯霉素的抗性橄榄油培养基上。在34℃倒置培养了6天。

2.3.2 再次对培养出来的菌种进行鉴定

2.4菌种的传代培养

2.4.1 将菌种稀释为四个浓度梯度用移液枪接种到编好号的培养基中,用涂布器 涂布均匀,放在34℃的恒温箱中进行培养,培养3天后,再一次进行传代培养。

2.4.2菌种的保存：斜面划线法进行接种培养，培养两天后放在4℃的冰箱里进 行保存备用。

2.5药品的提取及灭菌

2.5.1超声提取

2.5.1.1称取药品：何首乌、黄芩、三颗针和侧柏叶的粉末各5g放入塑料袋中

每个塑料袋中加入50ml的蒸馏水，并密封。

2.5.1.2超声提取：①接通电源；②调模式（MOOE）调时间（SET TIME）：20min；

调温度（SET TEMP）：50℃ ③温度到50℃后，把药品放入超声提取机中，进行提取。

2.5.1.3过滤：滤网+烧杯进行过滤

2.5.1.4离心：药液放入离心管中，配平，离心，离心后取上清液，放入药瓶中。

2.5.1.5巴氏消毒法进行灭菌：超声提取机中进行，调温度为60℃，把药品放入

其中，时间为30min。

2.5.1.6阳性药物的制备：阳性药品酮康唑用二甲基亚砜稀释，初始浓度为

40mg/ml；最适初始浓度为0.032mg/ml.用微孔过滤（微孔滤膜的孔径是0.45um,规格是Φ25m/m批号：2012年4月8日，生产厂家：上海兴亚净化材料厂）的方法来进行灭菌。

2.5.2煮沸法

称取药品：何首乌、黄芩、三颗针和侧柏叶各2g放入塑料袋中每个塑料袋

中加入10ml的蒸馏水，并密封。用沸水煮15min,过滤、离心;巴氏消毒法进行灭菌。

2.6接种:贴片法

2.6.1配制培养基

2.6.2药液的稀释： 把药液用无菌水稀释10倍，即初始浓度为10mg/ml(1ml

药液+9ml无菌水）；把稀释用的小药瓶分别编号：1，2，…10；1号加入1000ul的药液，2～10加入500ul的无菌水，自第一孔吸500ul的药液加入2号小药瓶中，混匀，并依次递倍稀释，每个小药瓶中放入3片滤纸片(直径R为6mm)进行浸泡，用一个小药瓶装500ul的蒸馏水并放入14片滤纸片(直径R为6mm)浸泡，用一个小药瓶(容量：5ml)装500ul的菌液并放入14片滤纸片浸泡，

2.6.3涂布： 一种药设置3个平行对照组，每个培养基上都做好标记并编号；用稀释好的菌液进行涂布，在涂布时注意用具的灭菌。共重复5次。

2.6.4贴滤纸片：在相应的培养基上对应着编号进行贴片。

2.6.5 培养:34℃培养72h，每天观察菌体生长情况，以菌体生长对照生长是否

良好为标准。

2.6.6结果判读：肉眼观察到菌体生长对照孔菌落生长良好，有菌落生长为无抑制作用，无菌落生长为有抑制作用。用游标卡尺来测量抑菌圈的大小。

2.7质控方法：每次试验的同时，以两种方法进行敏感性测定。每次结果显示其MIC总是介于同一范围，且各药物每次测定的MIC值波动不超过一个稀释度，方认为实验操作准确无误。

3结果

3.1头皮屑镜检结果：看到一些蓝紫色的点。孢子壁厚、单级出芽；初步判断为

马拉色菌。

3.2对培养出来的菌种进行鉴定：培养出了两种真菌菌落。

3.2.1 肉眼观察：

篇二：修辞学 马拉碦什中的三段论运用

As You See It

With the finishing of the University Entrance Examination, I have remembered the last class of mine in senior high school again. Different from others days, the days we all addicted to the book, the exercise, the discussion. We were so silent that even we can hear the breath of each other, but the voice from the man, who always called antique by us, is my head teacher.

Out of the window, the rain was washing the leaves; the wind was sweeping the top of tree, the birds up and down was seeking their nest. Inner the window, everyone lower their head, I guessed that some of them were weeping with the last song when I walked through your window sung by a girl. Listened the voice, my heart like a stone fall into water-plop, I cannot help myself from crying. Suddenly, the sound was interrupted by a rushed voice out of the door, the bell rang, and we had known that we had to finish the class.

A study of logos, pathos and ethos applied in “Marrakech”

For Aristotle, logic deals with universal and verifiable truth, while rhetoric deals with probable truth, that is, opinion and beliefs that can be advanced with greater or lesser certainty. As to the latter one, it called artistic proofs-fall within the province of art of rhetoric. Aristotle maintained that artistic proofs rely on three means of persuasion: logos, pathos, and ethos. He thinks that a persuasive essay must own that three means-logos, pathos and ethos. Refers to the Marrakech, Orwell successfully depicts the poverty of the inhabitants of Marrakech by describing the various aspects of their life to try to present a clear picture of the poverty of the people. To attach the purpose, in rhetoric, Orwell has a good use of the logos, pathos and ethos.

Logos, or rational appeal. It uses logical language and the relationship between cause and effect to appeal to the audience’s reasoning or understanding. That is to say, in Logos, it always by theory, concept, quotation or historical facts, authority views and related law to achieve the goal. There are two basic approaches people often use for logical reasoning: deduction and induction. About this point, we can see Orwell shows six almost uelated scenes, except for the fact that they demonstrate the

“invisibility” of the people in Morocco, which are: the corpse scene, the gazelle-Arabian scene, the Jew-cigarette scene, the landscape scene, the donkey-woman scene, and the bird-soldier scene. Orwell in deductive way to organize the essay, by the different scenes described, the author try to appeal the understanding of audience about the hard life in Marrakech. All the scenes were saw by the author self, which are objective descriptions and enforce the authentic. And among the essays, the large amount of quotation of the conversation from local people. Besides, Orwell wrote the whole passage in intensive logical structure and language-the transition words like “as”, “but “and “when” used in the beginning of some paragraphs. All above show the full use of Logos in the essay.

Pathos, or emotional appeal. It always related to emotion and sympathy. It usually arouses the strong affective feelings of the audience or reader to accept something ueason. Pathos refers to any affection, such as love, hatred, anger, ambition, fear, pity, jealously and sympathy, etc. People expressed Pathos by the language content to acknowledge others’ identity or strong emotion. Effective appeals to the audience’s emotion depend upon the skillful, and often witty, handling of language, frequently accompanied by exposure and eloquence. Exposure evokes moral indignation by condemning the unjust reality or revealing the difference between how things should be and how they. Eloquence refers to language used powerfully and fluently to appeal to people’s nobler emotions, like the sense of honor, love of one’s country and hometown, desire to reach toward virtue. Eloquence is often used emotively to express one’s emotions and evoke the sympathy of the audience on solemn occasions or issues of great significance. In Marrakech, Orwell used both of exposure and eloquence to describes the scenes utmost. Orwell tried to denounce the evils of colonialism or imperialism. He mercilessly exposes the poverty, misery and degradation of the native people in the colonies. These people are not considered nor are they treated as human beings. The cruel treatment the donkey receives evokes a greater feeling of sympathy in the breasts of the white masters than the miserable fate of the brown human beings. For example, to present the miserable life of the old women, there is such sentence “Then for the first time I noticed the poor old earth-colored bodies, bodies reduced to bones and leathery skin, bent double under the crushing weight”. Maybe Orwell is hyperbole in this situation, but when reader was caught the sight by this expression, they must have a great appeal for the sympathy. Another description of the corpse scene must have made someone feel sick even fear, but feel sympathy in the end. And the severely life of colonialism was written in this

way “What does Morocco mean to a Frenchman? An orange grove or a job in Government service. Or to an Englishman? Camels, castles, palm trees, Foreign Legionnaires, brass trays, and bandits.” So many examples like this in the essay. Orwell shows the poverty and miserable lives in this way to the reader, catch the emotion of reader properly and finish his purpose perfectly. The last should owe the narrative person is “I”, which gives the reader a true feeling to emerge into the circumstances.

Generally speaking, Marrakech is an excellent narrative works. A good works like that the use of pathos must occupy large ground to conceived the readers.

Ethos, or ethical appeal. It laid the foundation for the people’s credit. It makes others believe the reality of the language through the hero’s attitudes and qualities. In modern rhetoric, ethos refers to the personal standing, academic authority, and moral qualities of the writer, used to guarantee or raise the writer’s credibility and acceptability in the eyes of the readers. In the essay, Orwell presents it mainly through the attitudes I think. The attitudes to travel the area, to talk with the local people, to observe all the scenes cautiously for make the essay more reliable. The Orwell spent several months in Marrakech, Morocco from September 1938 to March 1939. The essay Marrakech must have been written during their stay here. Certainly, people never get to Morocco willing to believe what Orwell said mostly owed to the personnel experience. In Marrakech, Orwell also denounced the colonist, to some extent; they are the killer to kill the happiness of this land. Another reason why Orwell should have the deep feelings about the circumstance is that he once served with the Indian Imperial Police in Burma. He must saw the bad treatment of the people in India from the colonist. So without doubt, the author could write the essay which has a great appeal for the readers.

To sum up, Orwell describes objectively the suffering and misery of the colonial people in Marrakech, yet he manages to show that he is outraged at the spectacle of misery. He succeeds in imparting this feeling to his readers, first, through the clever choice of the scenes he describes; second, through the appropriate use of words; third, through the tone in which he describes these scenes; and finally, by contrasting the indignation at the cruel handling of the donkey with the unconcern towards the fate of the human being. All above, Orwell has a good proportion of logos, pathos, and ethos, especially the use of pathos to arouse the emotion of readers.

篇三：加拿大马拉斯比纳国际高中世界排名

学校介绍

马拉斯比纳国际高中（MIHS）是加拿大公立马拉斯比纳大学的附属中学，它是一所由民间投资、公立大学实施管理的高中，也是加拿大唯一的一所大学附中。马大附中提供10年级（高一）、11年级（高二）和12年级（高三）教育并授予BC省高中毕业统一文凭。马大附中位于加拿大西部温哥华岛上的海滨小城那奈莫Nanaimo市，距西部大都市温哥华乘轮渡仅1个半小时（乘快速轮渡仅40分钟）。马大附中坐落在马拉斯比纳大学校园内，它既有自用的教学环境，又与大学共享图书馆、计算机房、学生餐厅、剧场等很多设施。

据360教育集团介绍，国际学生进入马大附中，每年共有5次入学时间可供任选。班级的规模限制在15人。学校采用英语教学，入学前对学生的英语水平进行评估，对于英语未达标的学生，学校会安排先提高英语，然后逐步加入高中的课程。

马大附中国际毕业生去向

近20%：到欧洲读大学。80%以上留在北美深造，其中：升学率100%

约70%毕业生选择自动升入马大本科（部分学生三、四年级转入北美其它院校）其它30%左右学生进入其它院校：

英属哥伦比亚大学多伦多大学维多利亚大学西蒙弗雷泽大学

截止目前，留学360更新海外名校录取86789枚，其中哈佛大学43人、耶鲁大学56人、 斯坦福大学43人、麻省理工学院25人、牛津大学38人、剑桥大学35人、多伦多大学 290人、麦吉尔大学353人、悉尼大学1874人、墨尔本大学1286人、澳洲国立大学 1100人、香港大学120人、新加坡国立大学150人、南洋理工大学227人、新西兰奥克兰大学1241人。

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