伊犁黑蜂蜂胶对不同生长状态白假丝酵母菌抑菌作用的体外研究

目的探究伊犁黑蜂蜂胶对白假丝酵母菌游离状态及其生物膜状态的影响。方法采用NCCLS M27-A2酵母微量稀释法结合平板法测定蜂胶对游离状态下白假丝酵母菌的最小抑菌浓度(MIC)和最低杀菌浓度(MBC);玻片法在体外构建白假丝酵母菌生物膜模型,采用MTT法检测蜂胶对白假丝酵母菌生物膜状态下的最低生物膜清除浓度(MBEC)及抑制作用;激光共聚焦显微镜(CLSM)观察不同浓度蜂胶醇提取物对同一时间白假丝酵母菌生物膜的抑菌效果,并进行红蓝荧光染色定量分析。结果伊犁黑蜂蜂胶对浮游白假丝酵母菌的MIC为4mg/mL,MBC为8mg/mL;伊犁黑蜂蜂胶对生物膜状态下白假丝酵母菌的MBEC为16mg/mL,随着药物浓度的增加抑菌性逐渐增大,差异具有统计学意义(P0.05);CLSM观察,伊犁黑蜂蜂胶作用于白假丝酵母菌生物膜后可使生物膜内活菌比例降低,生物膜活性减弱,生物膜内实验菌死亡率随蜂胶浓度增加呈上升趋势,与阴性对照组比较差异有统计学意义(P0.05)。结论伊犁黑蜂蜂胶在体外对不同状态下的白假丝酵母菌均起到了明显的抑制作用。     

龙血素A对白色假丝酵母菌毒力因子ALS3、SAP4作用的体外研究

目的研究龙血素A对白色假丝酵母菌生物膜的抑菌作用,并初步研究龙血素A是否通过影响或干扰白色假丝酵母菌毒力因子的转录和表达实现的。方法建立白色假丝酵母菌生物膜体外模型,MTT法计算龙血素A对白色假丝酵母菌生物膜的抑制率;激光共聚焦观察龙血素A对不同时间白色假丝酵母菌生物膜的抑菌效果;RT-PCR技术检测药物作用下白色假丝酵母菌毒力因子表达水平的变化。结果随着药物浓度的增加,龙血素A对白色假丝酵母菌生物膜的抑菌作用增强。激光共聚焦观察显示龙血素A使白色假丝酵母菌生物膜内活菌死菌比例下降。RT-PCR结果表明龙血素A抑制了毒力因ALS3、SAP4的表达。结论龙血素A对白色假丝酵母菌生物膜有抑制作用,并在毒力因子ALS3、SAP4的表达过程中起到明显的抑制作用。     

乳杆菌发酵上清对耐药白假丝酵母生物膜的作用

目的研究德氏乳杆菌(Lactobacillus delbrueckii,L.delbrueckii)和发酵乳杆菌(Lactobacillus fermentum,L.fermentum)发酵上清液对3株耐氟康唑白假丝酵母临床分离株CA3、CA6、CA8生物膜形成和分散的作用。方法通过MIC试验,确认3株白假丝酵母临床分离株对氟康唑耐药;用96孔板构建体外白假丝酵母生物膜;用棋盘法分别检测L.delbrueckii、L.fermentum发酵上清液与氟康唑联用对3株白假丝酵母的作用;用XTT法对生物膜形成量进行定量分析;检测L.delbrueckii、L.fermentum发酵上清液与氟康唑联用对白假丝酵母时间-生长曲线的影响;显微镜拍照检测L.delbrueckii、L.fermentum发酵上清液单独和氟康唑联用对白假丝酵母生物膜形态的影响;平板培养法检测L.fermentum发酵上清液对白假丝酵母黏附作用的影响。结果 CA3、CA6、CA8三株临床分离菌株对氟康唑耐药,MIC浓度均为8μg/mL。L.delbrueckii与L.fermentum发酵上清液与氟康唑联用对3株耐药型白假丝酵母的生物膜形成与分散均未表现出协同作用,且发酵上清液与氟康唑联用效果不如发酵上清液单独处理效果好,L.fermentum发酵上清液对生物膜的分散作用较L.delbrueckii发酵上清液强。L.fermentum发酵上清液抑制白假丝酵母的初始黏附。结论 L.delbrueckii、L.fermentum发酵上清液单独应用时均对耐氟康唑白假丝酵母生物膜的形成与分散有干预作用,与氟康唑联用时未表现出协同作用,抑制耐药白假丝酵母生物膜的作用可能与抑制菌丝形成和起始黏附有关。     

假鹰爪鲜花挥发油成分研究

分别利用顶空固相微萃取法(Head Space-Solid phase microextraction,HS-SPME)和水蒸汽蒸馏法提取了黄色假鹰爪鲜花的挥发油成分,用气-质联用技术(GC-MS)对挥发油成分进行了分析,对比了两种提取方法得到的黄色假鹰爪鲜花挥发油成分的异同.顶空固相微萃取法得到的黄色假鹰爪鲜花挥发油成分中芳樟醇(46.19%),香茅醇(3.97%),反-香叶醇(4.21%),香叶酸甲酯(26.64%),(2Z)-3,7-二甲基-2,6-辛二烯酸乙酯(3.40%)五种物质的含量较高,其中三种醇类物质占总挥发油的54.37%;两种酯类物质占总挥发油的30.04%.水蒸汽蒸馏法得到的黄色假鹰爪鲜花挥发油成分中β-石竹烯(31.76%),α-石竹烯(8.63%),大根香叶烯D(4.65%),匙叶桉油烯醇(4.44%),α-石竹烯氧化物(12.98%)五种物质的含量较高.另外,通过对比发现不同生长期假鹰爪鲜花虽然香味差别较大,但是由水蒸汽蒸馏油得到的挥发油成分的种类及含量基本相同.     

溶菌酶对微小小单胞菌生物膜的影响

目的测定溶菌酶对微小小单胞菌及其生物膜的抑菌作用,并测出最小抑菌浓度(MIC)、最小杀菌浓度(MBC)和抑菌率。方法采用对倍稀释的方法,测定溶菌酶对微小小单胞菌的MIC、MBC;在96孔板中体外建立微小小单胞菌生物膜模型,采用MTT法检测溶菌酶对微小小单胞菌生物膜的影响;在六孔板中建立生物膜模型,使用激光共聚焦显微镜(CLSM)观察不同浓度溶菌酶对微小小单胞菌生物膜作用后的变化。结果溶菌酶对微小小单胞菌的MIC为0.0195 mg/mL,MBC为0.3125 mg/mL;CLSM观察结果显示,溶菌酶对微小小单胞菌生物膜的抑制作用随着浓度的增加而增强。结论溶菌酶对微小小单胞菌及其生物膜的生长和活性均具有抑制作用。     

丹皮酚对变形链球菌生物膜影响的体外研究

目的测定丹皮酚对变形链球菌的抑菌作用;共聚焦显微镜观察丹皮酚对变形链球菌生物膜结构和活性的影响。方法梯度法测定丹皮酚对变形链球菌的MIC(最小抑菌浓度)、MBC(最小杀菌浓度);体外构建变形链球菌生物膜模型,共聚焦显微镜观察不同浓度丹皮酚对变形链球菌生物膜作用后形态结构的影响并进行红绿荧光定量分析其活性变化。结果丹皮酚对变形链球菌的MIC为6.25mg/mL,MBC为25mg/mL;激光共聚焦显微镜观察丹皮酚对变形链球菌生物膜作用后其生物膜结构变稀疏,细菌链变短,生物膜活性也随丹皮酚浓度的提高而逐渐降低。结论丹皮酚对变形链球菌和变形链球菌生物膜结构及其活性均具抑制作用。     

棘托竹荪挥发油化学成分及抑菌作用的研究*

利用水蒸汽蒸馏法提取棘托竹荪的挥发油,得油率为0.45%。.应用气相色谱—质谱联用系统首次对其挥发油的化学成分进行研究。以FFAP柱分离出36个峰,用质谱法鉴定出28个成分,其主要成分为13-甲基-环氧十四烷-2-酮 (23.53%)、亚油酸(17.56%)、芹子烯(12.37%)、棕榈酸(8.20%)、9-十六碳烯酸(7.84%)、(-)-Lepidozenal(7.82%)等, 占总挥发油的97.76%。对挥发油进行抑菌试验,其结果为:桔黄青霉、啤酒酵母最敏感, 黑根霉、黑曲霉次之,白色假丝酵母、金色葡萄球菌,大肠杆菌稍差。     

大蒜素抗变异链球菌的致龋效果

目的研究大蒜素对口腔变异链球菌生长及其菌斑生物膜粘附的抑制作用。方法二倍稀释法梯度稀释测最小抑菌浓度(minimum inhibitory concentration,MIC),将MIC以上2个梯度浓度对应的培养物涂布于BHI培养基上进行次代培养获得最低杀菌浓度(minimum bactericidal concentration,MBC);酶标仪测A值观察不同浓度大蒜素抑菌效应;抑制产酸试验观察抑制细菌产酸效应;结晶紫法研究亚抑菌浓度提取物对变异链球菌粘附能力及生物膜总量的影响;采用激光共聚焦荧光显微镜(laser scanning confocal microscopy,LSCM)观察常态牙菌斑生物膜生长过程中及药物处理后牙菌斑生物膜中死菌和活菌的构成,研究其对牙菌斑生物膜结构和活性的影响。结果抑菌试验中,得到大蒜素MIC为12.8 mg/L,MBC为25.8 mg/L。MIC及亚抑菌浓度抑菌试验显示均有一定的抑菌性,抑制率为2.17%～67.12%,并且抑菌性与浓度梯度成正相关。产酸试验显示24 h内大蒜素明显抑制细菌产酸(P0.01),细菌粘附试验结果显示大蒜素在MIC时生物膜的生成速度最慢,生物膜的总量最低(P0.01)。共聚焦荧光显微镜可见大蒜素组随药物浓度增加,菌斑生物膜较薄,绿色的活菌及团块明显减少,抑制生物膜的生长。结论大蒜素对变异链球菌生长、产酸与粘附有一定抑制作用。     

妇科住院患者酵母样真菌感染的菌株分布与药敏研究

目的对妇科住院患者阴道分泌物酵母样真菌隐性感染的菌群分布与药敏结果进行回顾性分析,为临床围手术期合理使用抗真菌药物提供依据。方法妇科住院患者5813例阴道分泌物中分离到396株酵母样真菌,采用API-20Aux和ATBFUNGUS进行鉴定和药敏试验。结果396株酵母样真菌中,分离率最高的前3位是白色假丝酵母菌（59.6%）、光滑假丝酵母菌（21.2%）和接合假丝酵母菌（6.6%）。酵母样真菌对5-氟胞嘧啶（5-FC）、两性霉素B（AMB）和制霉菌素（NYS）敏感率最高,达90.7%、92.7%和96.2%,咪康唑（MIC）、益康唑（ECO）和酮康唑（KET）分别为74.2%、61.6%和69.4%。结论住院妇科患者阴道分泌物酵母样真菌隐性感染主要以白色念珠菌和光滑念珠菌为主,感染的酵母样真菌对5-FC、AMB和NYS的药物敏感性较高,对唑类抗真菌药则有不同程度的耐药率。     

金诺芬对表皮葡萄球菌及其生物膜的影响

目的研究金诺芬对表皮葡萄球菌及其生物膜的作用。方法利用微量稀释法药敏试验检测金诺芬对表皮葡萄球菌浮游菌生长的影响,并利用结晶紫染色和激光共聚焦显微镜观察金诺芬对表皮葡萄球菌生物膜形成的影响。结果金诺芬对表皮葡萄球菌的MIC和MBC分别为0.125～0.250μg/mL和2.000～4.000μg/mL。同时,4μg/mL金诺芬还能显著抑制表皮葡萄球菌标准菌株和临床菌株生物膜的形成(P0.05)。通过激光共聚焦显微镜观察发现金诺芬能有效抑制生物膜的形成,降低生物膜的聚集,并增加死亡细菌的比例。结论金诺芬能显著抑制表皮葡萄球菌浮游菌的增殖和生物膜的形成。     

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.     

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.     

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.     

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.     

Interconnection of parameters of regulation system of lipid peroxidation and of morphophysiological parameters of mouse liver

A complex analysis of seasonal fluctuations of the mean group parameters of the system of regulation of lipid peroxidation has been performed in liver of Balb/c mice. Association of lipid characteristics and morphophysiological parameters is studied in the Balb/c mouse liver. An inter-connection is revealed between the liver index and the amount of lysoforms of phospholipids, the scale and character of the interconnection differing essentially depending on proportion of phos-phatidylcholine in mouse liver phospholipids.     

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。     

Physiological characteristics of various species of strains of carboxydobacteria

Seven strains of aerobic carbon monoxide-oxidizing bacteria (carboxydebacteria) when growing on CO as sole source of carbon and energy had doubling times which ranged from 12–42 h. The activity profiles obtained after discontinuous sucrose density gradient centrifugation indicated that the CO-oxidizing enzymes are soluble and the hydrogenases are membrane-bound in all strains examined. The CO-oxidizing enzymes of Pseudomonas carboxydohydrogena, Pseudomonas carboxydoflava, Comamonas compransoris, and the so far unidentified strains OM2, OM3, and OM4 had a molecular weight of 230,000; that of Achromobacter carboxydus amounted to 170,000. The molecular weights of the CO-oxidizing and H₂-oxidizing enzymes turned out to be identical. The cell sonicates were shown to catalyze the oxidation of both CO and H₂ with methylene blue, thionine, phenazine methosulfate, toluylene blue, dichlorophenolindophenol, cytochrome c or ferricyanide as electron acceptors. Methyl viologen, benzyl viologen, FAD⁺, FMN⁺, and NAD(P)⁺ were not reduced. The spectrum of electron acceptors was identical for all strains tested. Neither free formate, hydrogen nor oxygen gas were involved in the CO-oxidation reaction. Methylene blue was reduced by CO at a 1:1 molar ratio. The results indicate that CO-oxidation by carboxydobacteria is catalyzed by identical or similar enzymes and that the reaction obeys the equation CO+H₂OCO₂+2H⁺+2e^- as previously shown for Pseudomonas carboxydovorans.Dedicated to Otto Kandler remembering almost three decades of enjoyable cooperation     

Modulation of allostery of pyruvate kinase by shifting of an ensemble of microstates

Since the introduction of the concepts of allostery about four decades ago, much advancement has been made in elucidating the structure-function correlation in allostery. However, there are still a number of issues that remain unresolved. In this review we used mammalian pyruvate kinase (PK) as a model system to understand the role of protein dynamics in modulating cooperativity. PK has a triosephosphate isomerase (TIM)(α/β)₈ barrel structural motif. PK is an ideal system to address basic questions regarding regulatory mechanisms about this common (α/β)₈ structural motif. The simplest model accounting for all of the solution thermodynamic and kinetic data on ligand-enzyme interactions involves two conformational states, inactive E^T and active E^R. These conformational states are represented by domain movements. Further studies provide the first evidence for a differential effect of ligand binding on the dynamics of the structural elements, not major secondary structural changes. These data are consistent with our model that allosteric regulation of PK is the consequence of perturbation of the distribution of an ensemble of states in which the inactive E^T and active E^R represent the two extreme end states. Sequence differences and ligands can modulate the distribution of states leading to alterations of functions. The future work includes: defining the network of functionally connected residues; elucidating the chemical principles governing the sequence differences which affect functions; and probing the nature of mutations on the stability of the secondary structural elements, which in turn modulate allostery.