沈阳地区铜绿假单胞菌药物敏感性测定及质粒图谱分析

目的 测定铜绿假单胞菌对22种药物的敏感性,帮助临床选择用药。并对分离株进行质粒图谱分析以了解耐药菌株的流行情况。方法 药物敏感性实验采用纸片琼脂扩散法,质粒指纹图谱分析采用碱变性法提取质粒DNA,限制性内切酶切割后进行凝胶电泳分析。结果 铜绿假单胞菌对头胞哌酮、氧派酸、丁胺卡那霉素、壮观霉素、多粘菌毒和头孢三嗪的敏感率在84％－100％之间。所有菌株对其他16种抗性素均有不同程度的耐药。质粒DNA图谱分析显示,12株被检测菌株中有11株含有质粒DNA,其中8株含有23kb质粒DNA。结论 铜绿假单胞菌对头胞哌铜、氟派酸、丁胺卡那霉素、壮观霉素、多粘菌素敏感;多数耐药菌株含23kb质粒DNA。     

嗜麦芽假单胞菌酪氨酸酶基因在类产碱假单胞菌中的转移与表达

类产碱假单胞菌 (Pseudomonaspseudoalcaligenes)是从盐城市农村粪池中采集的自然病死蝇蛆体内分离的具有显著杀蛆作用的细菌 ,野外使用易受阳光中紫外线的影响而失活。黑色素具有很强的抗辐射作用。将构建的含有嗜麦芽假单胞菌酪氨酸酶基因的质粒pWSY导入类产碱假单胞菌体内 ,使后者获得了稳定产生黑色素的能力。Southern杂交实验证实酪氨酸酶基因来源于嗜麦芽假单胞菌。SDS PAGE电泳显示该重组子体内额外表达了一分子量约为 1 8kD的蛋白 ,该蛋白很可能就是重组子表达的酪氨酸酶。经测定 ,重组子抗辐射作用明显增强 ,有效杀蛆时间显著延长 ,对畜、禽安全     

ICU感染嗜麦芽窄食假单胞菌分布与耐药性分析

目的了解ICU感染嗜麦芽窄食假单胞菌的分布与耐药性,比较本院ICU与非ICU嗜麦芽窄食假单胞菌的耐药性,为临床合理应用抗菌药物提供科学依据。方法对2011年6月-2014年6月临床标本中分离的嗜麦芽窄食假单胞菌120株采用K-B纸片法进行药敏试验,应用SPSS 20.0对数据进行统计分析。结果嗜麦芽窄食假单胞菌主要来源于呼吸道标本(痰),占69.17%。临床分布以ICU多见,占30.00%。在20种抗菌药物中有14种抗菌药物耐药率50%,仅氧氟沙星、复方磺胺甲唑、米诺环素、左氧氟沙星等有较低耐药率,耐药率分别为0.00%、16.67%、1.67%、9.17%;头孢噻肟、阿莫西林/克拉维酸、亚胺培南、头孢哌酮/舒巴坦、美罗培南等12种抗菌药物ICU与非ICU耐药率分别为100.00%,66.67%;97.22%,75.00%;100.00%,40.48%;97.22%,33.33%;100.00%,79.76%;差异有统计学意义(P0.05)。结论临床感染嗜麦芽窄食假单胞菌对常用抗菌药物耐药严重,ICU嗜麦芽窄食假单胞菌耐药率明显高于非ICU,临床应高度重视,控制感染。     

嗜麦芽假单胞菌对藻胶的降解作用

许多学者认为嗜麦芽假单胞菌(Pseudomonas maltophilia)是一种与医院感染有关的机会致病菌,但它广泛地存在于自然界,而它在其中的作用仍不清楚。研究嗜麦芽假单胞菌对藻胶的降解作用,不仅为了阐明该菌的某些生态学特性,尤其是它在自然界物质循环中的作用,有理论上的意义,而且在生产实践     

嗜麦芽假单胞菌的碱性蛋白酶基因在大肠杆菌中的克隆与表达

用pUCl8质粒作为载体,将嗜麦芽假单胞菌(Psedeomonas maltophilia P27)的碱性蛋白酶基因克隆到大肠杆菌(E．Coli TGI)中,得到3株能分泌碱性蛋白酶的阳性克隆G1,G2和G3。其中G3所分泌的碱性蛋白酶活性最高,大约是出发菌株的3—4倍,对3株阳性克隆所含的重组质粒psJl,psJ2和psJ3进行限制酶酶切分析表明,酶活最高的阳性克隆G3所含的重组质粒psJ3的插入片段最小,大约是2．8kb;其它两株的重组质粒pSJl和pSJ2含有同样大小的插入片段,约为5．5kb。     

GL－7－ACA酰化酶基因克隆及在大肠杆菌中的表达

本文报道了利用BamHⅠ酶解的载体pBR322 DNA与Sau3A部分酶解的假单胞菌Ps．130染色体DNA片段构建重组质粒,并用³²P标记的寡核苷酸探针从3205个含有外源片段的重组质粒中检测得到7株阳性克隆株。进一步经¹²⁵Ⅰ标记的抗体／抗原放射免疫反应、GL－7－ACA酰化酶活力测定以及酶反应产物的纸谱色层分析,由上述7株阳性株中鉴定出3株能在大肠杆菌中表达酰化酶活力的基因克隆株。对该3株菌的重组质粒——pMR 5、pMR 6和pMR-7-DNA的初步凝胶电泳分析表明,pMR 5和pMR 7质粒中插入片段的分子量为6．8kb,质粒pMR 6则带有5．7kb的外源片段。实验还比较了重组质粒pMR 5在8株不同的大肠杆菌宿主菌中,GL－7－ACA酰化酶的表达水平。     

酿酒酵母两种不同嗜杀菌株的比较研究

以酿酒酵母两种不同类型的嗜杀菌株SK4（K1型）和ERRI（K2型）为材料,分析了不同嗜杀酵母的嗜杀特性,两株嗜杀酵母具有相互杀死作用,其嗜杀活性与菌体生长有关。SK4和ERRI的嗜杀质粒的比较表明：M1－dsRNA质粒和M2-dsRNA质粒分子量分别为1．7kb和1．5kb,两株菌的L－dsRNA质粒均为4．0kb。用高温和紫外线处理嗜杀酵母,嗜杀活性随之消失,消除菌中的M－dsRNA质粒也相应消失,嗜杀活性的消除率随菌株和消除剂的不同而变化。实验证明两株菌产生的毒素蛋白的最适嗜杀作用条件不同,最适pH和温度分别为4．8、16℃和4．0、22℃,但两种毒素蛋白均对在对数生长期的敏感细胞作用最显著。     

马齿苋提取液对铜绿假单胞菌R质粒消除作用的研究

据报道,铜绿假单胞菌R质粒携带者达叨．24％［1］,这是该菌对多种抗生素多重耐药的重要物质基础、用人工方法使R质粒从宿主菌中消除,使其恢复对抗生素的敏感性,是临床治疗多重耐药菌感染的新思路。本实验用马齿觅提取液做消除剂,对铜绿假单胞菌P29株R质粒进行体内、体外消除试验,现报告如下。1材料与方法1．1材料1．1．1菌种铜绿假单胞菌P29株（本室保存）,从长春市烧伤医院患者分离并鉴定。1．1．2马齿觉提取液马齿觅购手吉林大药房,由我校药物研究室李平亚副教授鉴定。按文献［2］方法提取。1．1．3试剂GM、KM、SM、TC均为…     

2007年至2009年3种非发酵革兰阴性杆菌耐药性分析

目的研究自临床分离的铜绿假单胞菌、鲍曼不动杆菌、嗜麦芽寡养单胞菌等3种非发酵革兰阴性杆菌的耐药性及其变迁,为合理使用抗菌药物提供依据。方法临床标本按常规方法分离培养,用生物梅里埃API分析系统鉴定细菌种类,根据CLSI规定的标准,采用纸片扩散法进行体外药敏测定,WHONET软件进行统计分析。结果 2007年至2009年共分离到铜绿假单胞菌310株、鲍曼不动杆菌280株和嗜麦芽寡养单胞菌86株。其中亚胺培南不敏感的铜绿假单胞菌为32.5%,亚胺培南不敏感的铜绿假单胞菌对其他药物的耐药率也明显高于亚胺培南敏感菌株;鲍曼不动杆菌对大部分抗生素的耐药率都高于60%,对亚胺培南、美罗培南3年总的敏感率为51.6%和48.2%;嗜麦芽寡养单胞菌对米诺环素的敏感率为93.8%。比较3年耐药率的变迁,2009年部分抗菌药物耐药性增长的势头得到遏制。结论非发酵菌是引起院内感染的重要病原菌,对多种抗菌药物都有较高的耐药性,合理使用抗生素,加强医院感染的控制对减少细菌耐药性的产生有非常重要的作用。     

质粒pNK866的拷贝数测定和限制酶图谱

假单胞菌(Pseudomonds)降解质粒的基因组织和表达调控的研究,以及以这些质粒为基础的高效降解环境污染物菌株和产生精细化工产品菌株的构建,是当前一个极为活跃的研究领域.但是,由于假单胞菌基因在大肠杆菌中的表达水平非常低,可用的假单胞菌载体又比较少,使这方面的研究遇到不少困难.目前使用较多的是来自质粒RSF1010的载体,但由于分子量较大,使用不方便,因此构建新的假单胞菌载体是该领域研究的一项重要内容.我们从弯曲假单胞菌(P.geniculata)AS 1.866中分离到质粒pNK866.本文报道了该质粒的拷贝数和限制酶图谱,为假单胞菌新载体的构建奠定了基础.     

六种藓类植物茎的比较解剖学研究

通过对6种藓类植物,即褶叶青藓(Brachythecium salebrosum(Web.et Mohr.)B.S.G.)、湿地匐灯藓(Plagiomnium acutum(Lindb.)Kop.)、侧枝匐灯藓(Plagiomnium maximoviczii(Lindb.)Kop.)、大凤尾藓(Fissidensnobilis Griff.)、大羽藓(Thuidium cymbifolium(Doz.et Molk.)B.S.G.)和大灰藓(Hypnum plumaeforme Wils.)嫩茎和老茎的石蜡切片和显微观察发现,同一藓类植株的嫩茎和老茎,茎结构稳定,不同种藓类植物茎横切面具有不同特征.植物体茎横切面形状、表层细胞的层数、细胞大小和细胞壁厚薄、皮层细胞大小和形状、中轴的有无以及比例等特征可以作为藓类植物的分科分类依据之一.     

Cause of Senescence of Nine Sorts of Flowers

The levels of endogenous phytohormones and respiratory rate in nine sorts of flowers such as Cymbidium faberi Rolfe, Nopalxochia ackermannii Kunth and others were investigated both at full bloom and senescence and meanwhile the effect of exogenous phytohormones on prolonging the blossoms and promoting ethylene production were tested. There is a high content of endogenous ethylene in all the long-lived flowere, about 3–16 folds higer than the short-lived ones. There is a high level of ABA at full blooming flowers of short-lived flowers, in which there is no or only some cytokinins in it, but the ratio of CTK (6BA+zeatin)/ABA is smaller(l.7). The endogenous ABA reached a much higher level at senescence in all nine sorts of flowers, so it is reasonable to consider that it is ABA which plays an important role of regulation in controlling flower's senescence. There is a much higher level of GA3 and zeatin in the long-lived flowers which is not demonstrated in the shortlived ones. The respiratory rate is one of the factors controtling the longevity of flowers, but it does not play a decided role. Application of 6BA and zeatin prolongs distinctly orchid’s longevity, however exogenous IAA through the promotive action on ethylene production, evidently extends the longevity of the flowers of the Nopalxochia ackermannii Kunth.     

龙胆科药用植物化学成分的研究现状

龙胆科植物在我国的分布范围很广,且多数为药用植物,其多数种属的药用植物,至今其化学成分尚未被系统研究。综述了目前龙胆科药用植物的化学成分的研究现状及一般提取方法,对近年来发现的环烯醚萜及裂环烯醚萜类化合物进行了总结,为本科药用植物的更深入研究提供了参考。     

The effects of glutamine of the maintenance of embryogenic cultures of Cryptomeria japonica

Summary Embryogenic tissues of sugi (Cryptomeria japonica) were induced on a modified Campbell and Durzan (CD) medium containing 1 μM 2,4-dichlorophenoxyacetic acid (2,4-D) and 600 mg l⁻¹ glutamine, and subcultured in the medium of the same composition for over 1 yr. This resulted in a mixed culture of embryogenic and non-embryogenic cells. When embryogenic cells were isolated and cultured independently, their capacity to form embryogenic aggregates was lost. Thus, the non-embryogenic cells present within a mixed culture system were essential to the formation of embryogenic aggregates. When embryogenic tissues were isolated and cultured independently on a high glutamine-containing (2400 mg l⁻¹) medium, dry weights and endogenous levels of glutamine increased, and the tissue could generate a large number of embryogenic aggregates. Amino acid analysis of embryogenic and non-embryogenic cells from the maintenance culture indicated a higher level of glutamine was present in the latter. The high endogenous level of glutamine in the non-embryogenic portion of mixed cell masses may be the supplier of glutamine for maintaining the embryogenic property of the tissues.     

Specificity of action of piperidylcholine derivatives as substrates of cholinesterases of various origin

The review deals with study of enzymologic properties of a novel highly specific acetylcholinesterase substrate, N-(β-acetoxyethyl) piperidinium iodomethylate (“piperidylcholine”), and its 30 derivatives that were tested as effectors of cholinesterases of mammals and various species of Pacific squids. It was proven for the first time that responsible for specificity of action was structure of cyclic ammonium grouping of the alcohol part of molecule of the ester substrate. Analysis of specificity is performed based on enzymatic hydrolysis parameters—activity of catalytic center of cholinesterases and bimolecular constant of the reaction rate that are determined at optimal and low substrate concentrations. Among the specially synthesized group of thioester compounds there is revealed one more highly specific acetylcholinesterase substrate—N-(β-acetoxyethyl) piperidinium.     

真菌类遗传学分析的知识结构教学

本文以认知结构理论为指导,讨论了真菌类遗传分析与高等动植物遗传分析的内在联系,认为利用这种内在联系进行教学可收到好的效果并说明了作者的具体教学过程。 Abstract:In the paper, the relationship between genetic analysis of Fungi and genetic analysis of high animal and plant was discussed.A good results were obtained when we adopted this method in the teaching.     

The mechanism of hatching of eggs of Haemonchus contortus

Fluid collected from hatching eggs of Haemonchus contortus contained a lipase which hydrolysed 2-naphthyl laurate (about 0·7 μmol naphthol freed /h/10⁶ eggs). The fluid also hydrolysed l-leucinamide (about 2·3 μmol leucine freed/h/10⁶ eggs). The fluid when added to normal or heated eggs caused ‘hatching’. ‘Hatching’ also occurred in exsheathing fluid from infective juveniles and in a preparation of pancreatic lipase containing leucine aminopeptidase. A purified mammalian leucine aminopeptidase in combination with several different lipases did not attack egg shells.The ‘spontaneous’ hatching of eggs of H. contortus was strongly inhibited by 1,10-phenanthroline, 10^?3M, and this inhibition was reversed by Zn²⁺. However, the inhibition of ‘hatching’ of eggs in externally applied hatching fluid, or the hydrolysis of leucinamide in hatching fluid was generally less marked.     

Photoregulation of seed germination of wild-type and of an aurea-mutant of tomato

Seed germination of an aurea mutant of tomato ( Lycopersicon esculentum Mill.) is promoted by continuous irradiation with red, far-red or long-wavelength far-red (758 nm) light as well as by cyclic irradiations (5 min red or 5 min far-red/25 min darkness). Far-red light applied immediately after each red does not change the germination behaviour. Seed germination of the isogenic wild-type, cv. UC-105, is promoted by continuous and cyclic red light while it is inhibited by continuous and cyclic far-red light and by continious 758 nm irradiation. Far-red irradiation reverses almost completely the promoting effect of red light. The promoting effect (in the aurea mutant) and the inhibitory effect (in the wild-type) of continuous far-red light do not show photon fluence rate dependency above 20 nmol m⁻² s⁻¹. It is concluded that phytochrome controls tomato seed germination throgh low energy responses in both the wild type and the au mutant. The promoting effect of continuous and cyclic far-red light in the au mutant can be attributed to a greater sensitivity to P_fr.