表皮葡萄球菌生物被膜对红霉素渗透性的研究

为探讨表皮葡萄球菌生物被膜对红霉素的渗透性,我们采用生物被膜抗生素渗透模型检测Staphylcoccus epidermidis 1457、1457-msrA和临床分离株S68生物被膜不同时间点红霉素的渗透率,并用吖啶橙染色激光共聚焦显微镜观察生物被膜内细菌RNA/DNA的相对含量;扫描电子显微镜观察膜内细菌的密度.红霉素作用36 h后,1457、1457-msrA、S68的渗透率分别为0.93,0.55和0.4;1457渗透地较快,8 h后渗透率即达到0.58,而1457msrA和$68相对较为缓慢,24 h后分别为0.499和0.31:吖啶橙染色可见红霉素作用下膜内菌RNA和DNA的相对比例减小,生长速率下降;扫描电子显微镜观察可见生物被膜红霉素作用后空气面的细菌数与琼脂面相比均较少,细胞碎片相对较多,而对照组(无抗生素作用)琼脂面和空气面的细菌密度和分布较均匀.可见红霉素可渗透入表葡茵生物被膜,但不能完全杀死膜内细菌;膜内细菌在生物被膜环境中生长速率下降,有助于降低细菌对红霉素的敏感性.     

抗菌肽17BIPHE2对金黄色葡萄球菌生物被膜的抑制作用

【目的】研究抗菌肽17BIPHE2单独使用及联合抗生素对金黄色葡萄球菌(Staphylococcus aureus)生物被膜的抑制作用。【方法】采用刚果红平板测试法和结晶紫染色评估受试菌形成生物被膜的能力;微量肉汤稀释法和琼脂平板测试法测定金黄色葡萄球菌最小抑菌浓度(MIC)和最小杀菌浓度(MBC);利用抑制金黄色葡萄球菌黏附实验和生物被膜形成抑制实验观察17BIPHE2单独使用及联合抗生素对生物被膜黏附阶段和形成阶段的影响;通过扫描电子显微镜(SEM)观察17BIPHE2单独使用及联合抗生素对成熟生物被膜的清除作用。【结果】17BIPHE2的MIC为8μmol/L,1/2×MIC就可以有效抑制浮游菌的生长。单独使用17BIPHE2在细菌黏附阶段抑制率为40%,在生物被膜形成阶段抑制率达到35%。17BIPHE2联合抗生素使用较单独使用抗生素其抑制率均有所下降。生物被膜成熟阶段17BIPHE2于1/4×MIC浓度即可促进生物被膜崩解,1×MIC生物被膜崩解同时细菌黏附量有所下降,联合万古霉素促进生物被膜崩解同时细菌胞质大量外泄。【结论】抗菌肽17BIPHE2具有良好的抑制金黄色葡萄球菌生物被膜作用,联合抗生素其抗生物被膜作用进一步提高。这将为治疗由金黄色葡萄球菌生物被膜引起的相关感染提供了一个新思路。     

苦参碱、鱼腥草素钠及其与红霉素联用对表皮葡萄球菌生物被膜渗透作用的研究

目的比较苦参碱、鱼腥草素钠及其与红霉素联用对表皮葡萄球菌生物被膜的渗透性,为中药及中西药联用治疗表皮葡萄球菌引起的相关感染提供依据。方法在体外建立SE-BF(表皮葡萄球菌生物被膜模型),PB(生物被膜渗透模型);观察并测量含药滤纸片抑菌圈直径(每个时间点取样3个),计算苦参碱、鱼腥草素钠及其与红霉素联用对PB的渗透率。结果苦参碱、鱼腥草素钠、苦参碱与红霉素及鱼腥草素钠与红霉素联用对Se738的36h渗透率分别达到93.94%、96.97%、89.34%和93.53%;苦参碱、鱼腥草素钠、苦参碱与红霉素及鱼腥草素钠与红霉素联用对ATCC 35984的36h渗透率分别达到82.83%、88.55%、91.47%和96.76%。结论苦参碱、鱼腥草素钠及其与红霉素联用,对表皮葡萄球菌生物被膜均有较强的渗透性。     

铜绿假单胞菌生物被膜对抗菌药物作用影响的实验研究

细菌生物被膜形成在难治性感染中起重要作用,本实验在体外进行铜绿假单胞菌生物被膜与抗菌药物作用的研究。铜绿假单胞菌在生理盐水特氟隆系统孵育6d,可在特氟隆表面上形成细菌生物被膜(Biofilm),经扫描电镜证实,它包裹在微菌落表面。应用环丙氟哌酸,甲红霉素,罗红霉素,穿心莲作用于生物被膜细菌并观察结果。发现与2倍MIC环丙氟哌酸作用4h后,悬浮细菌存活率降至002%,但生物被膜细菌存活率则为41%,当10μg/mL甲红霉素,12μg/mL罗红霉素,005g/mL穿心莲分别与环丙氟哌酸联合应用时,生物膜细菌存活率分别降至02%,05%和27%。结果表明生物被膜细菌对环丙氟哌酸的抵抗力较悬浮细菌明显增强,而环丙氟哌酸分别与甲红霉素,罗红霉素,穿心莲联合应用则有明显抑菌作用。推测此种联合用药方式可能提高对难治性感染的治疗效果     

铁基纳米酶对鼠伤寒沙门菌生物被膜的影响

运用结晶紫染色定量法、生物被膜形态观察、生物被膜干重称量法、活菌定量计数法和细菌内活性氧检测法,评估氧化铁纳米酶和硫化铁纳米酶对鼠伤寒沙门菌生物被膜的影响及其机制.结果显示:鼠伤寒沙门菌S025株与这两类铁基纳米酶共孵育48h后,其生物被膜结晶紫染色吸光度值(A)、生物被膜厚度、生物被膜干重和活菌数量与未处理组相比均显著下降,活性氧水平显著上升,其中硫化铁纳米酶效果优于四氧化三铁纳米酶;在生物被膜形成后,加入铁基纳米酶处理0.5h、2h和12h,生物被膜结晶紫染色A值、生物被膜厚度、生物被膜干重和活菌数量与未处理组相比均显著下降,活性氧水平显著上升,硫化铁纳米酶效果同样优于四氧化三铁纳米酶.以上结果表明,铁基纳米酶通过调控鼠伤寒沙门菌胞内活性氧水平,不仅可以预防该菌的生物被膜形成,而且可以破坏已形成的生物被膜,本研究将有助于预防和治疗鼠伤寒沙门菌生物被膜引起的相关疾病.     

黄芩素抑制金黄色葡萄球菌生物被膜的形成

细菌生物被膜的形成是导致细菌耐药和引起持续性感染的主要原因之一。本文通过检测黄芩素对金黄色葡萄球菌26112菌株（Staphylococcus aureus 26112,SA26112）多糖细胞间黏附素（polysaccharide intercellular adhesion, PIA）的合成和胞外DNA（extracellular DNA,eDNA）释放量的影响,及其对icaA和cidA基因表达量的影响,探讨黄芩素对金黄色葡萄菌生物被膜形成的抑制作用及其机制。结果显示,黄芩素能抑制SA26112生物被膜的形成,其抑杀SA26112的最低抑菌浓度和最低杀菌浓度均为0.04 mg/mL。0.16 mg/mL黄芩素和256 μg/mL环丙沙星单独作用时,均不能杀死其成熟生物被膜内的SA26112细菌,而当二者联用时则可杀死成熟生物被膜内的细菌。黄芩素能显著抑制SA26112菌株PIA的合成、eDNA的释放量及icaA和cidA基因的相对表达量。其中,0.04 mg/mL黄芩素作用SA26112菌株24 h,与对照组相比,eDNA的释放量减少97%,icaA和cidA基因的相对表达量分别减少62%和41%。上述结果表明,黄芩素能抑制SA26112菌株生物被膜的形成,其作用机制可通过降低icaA和cidA的基因表达量,进而影响PIA的合成和eDNA的释放,来抑制金黄色葡萄球菌生物被膜的形成。     

噬菌体及其裂解酶对细菌生物被膜作用的研究进展

细菌形成的生物被膜,可保护细菌不易被抗生素杀死,这给临床上相应疾病的治疗及医疗器械的消毒带来极大困难。研究表明,噬菌体及其裂解酶对生物被膜有降解作用。噬菌体能清除细菌在有生物活性或无生物活性的介质表面形成的生物被膜。此外,噬菌体裂解酶比如LySMP、肽酶CHAPk、细胞壁溶解酶CWHs等能清除特定的生物被膜,这可能与裂解酶直接溶菌和裂解细菌细胞外基质有关。同时,与抗生素、钴离子、氯等物质联合使用时,噬菌体对生物被膜的清除作用会更强。本文从噬菌体、噬菌体编码的裂解酶、以及它们联合其他物质对细菌生物被膜的作用进行综述,并对其实际应用做了展望。     

十二烷基苯磺酸钠对引发内置物感染表皮葡萄球菌生物被膜的作用

【背景】随着医用内置物的广泛使用,由表皮葡萄球菌生物被膜导致的医院获得性感染不断增多,目前鲜见关于表面活性剂针对表皮葡萄球菌生物被膜作用的报道。【目的】通过研究阴离子型表面活性剂十二烷基苯磺酸钠(sodium dodecyl benzene sulfonate,SDBS)分别对ATCC 35984 (产膜表皮葡萄球菌标准株)生物被膜的清除、生物被膜内细菌代谢和形成生物被膜的关键物质多糖胞间黏附素(polysaccharide intercellular adhesion,PIA)产生的影响,为临床使用SDBS防治由表皮葡萄球菌生物被膜引起的相关感染提供可靠的理论及实践依据。【方法】利用XTT减低法,评价SDBS对ATCC 35984已形成生物被膜的清除效率及对生物被膜内细菌代谢的影响;激光共聚焦显微镜观察SDBS对生物被膜作用的效果;采用刚果红培养基观察SDBS对PIA产生的影响。【结果】浓度为256、128、64、32、16 mg/L的SDBS在作用6、12、24 h时,对ATCC 35984的生物被膜均有显著的清除效率(P0.01);浓度为32 mg/L时对生物被膜内细菌的代谢有显著抑制作用(P0.05),并随作用浓度的增加而增强;激光共聚焦显微镜观察显示256、128、64 mg/L的SDBS对生物被膜的清除效率较为理想,SDBS浓度为64、32 mg/L时对PIA的形成无明显抑制作用。【结论】SDBS对表皮葡萄球菌生物被膜内细菌的代谢有显著抑制作用,对生物被膜形态结构有显著破坏作用。     

植物提取物及其活性成分抑制细菌生物被膜的研究进展北大核心CSCD

大量研究报道生物被膜细菌对抗生素的耐药性是浮游菌的10–1 000倍,据报道细菌生物被膜是80%以上细菌感染的罪魁祸首,对医疗保健领域构成了严峻的挑战。植物提取物及其活性成分对细菌生物被膜有明显的抑制作用,包括减少生物被膜量、生物被膜活菌数以及清除已经成熟的生物被膜等。该文对这些有效的植物提取物及其活性成分进行了总结,并分析了其抗细菌生物被膜的作用机制。旨在为防治细菌生物被膜感染的植物类药物的开发提供参考。     

波罗的海希瓦氏菌AI-2/LuxS对生物被膜和致腐的调控

【目的】波罗的海希瓦氏菌(Shewanella baltica)是冷藏海产鱼类的特定腐败菌。研究群体感应信号AI-2/Lux S对鱼源S.baltica生物被膜和致腐的调控作用。【方法】扩增SB11分离株的lux S基因,用自杀性质粒构建lux S基因缺失株,通过结晶紫染色、珠涡流法、显微镜观察和HPLC,比较分析野生株与缺失株△lux S在4°C和28°C下生物被膜形成、粘附能力、泳动性和致腐产物的差异。【结果】S.baltica SB11中扩增获得lux S基因,生物信息学分析显示Lux S蛋白由169个氨基酸构成,含有保守的His-Thr-Leu-Glu-His(HTLEH)模体和关键氨基酸位点,蛋白三维空间结构与其他细菌相似。与野生株相比,△lux S缺失株上清荧光信号散失,但不影响生长,生物被膜形成期和成熟期的含量显著低于野生株,在4°C培养96 h和28°C培养24 h被膜分别减少20.1%和27.9%。缺失株在不锈钢片的粘附能力明显减弱,其中在4°C培养72 h和28°C培养24 h后的粘附量比野生株分别减少6.48%和6.57%。荧光显微镜观察发现,野生株能快速粘附于玻璃片,聚集形成大量生物被膜,而△lux S仅形成平坦稀疏的被膜,粘附细菌降低,CLSM证实野生株和△lux S的成熟被膜厚度分别为68.95μm和36.44μm。并且,△lux S株在4°C和28°C下泳动性均显著强于野生株。然而,野生株和△lux S株三甲胺和腐胺积累无差异。【结论】鱼源S.baltica中Lux S蛋白保守,AI-2/Lux S参与被膜、粘附能力及泳动性等多种生物被膜形成相关的调控作用,然而不是该菌致腐能力的功能性群体感应信号。     

Recent advances in the study of Kaposi's sarcoma-associated herpesvirus replication and pathogenesis

It has now been over twenty years since a novel herpesviral genome was identified in Kaposi's sarcoma biopsies. Since then, the cumulative research effort by molecular biologists, virologists, clinicians, and epidemiologists alike has led to the extensive characterization of this tumor virus, Kaposi's sarcoma-associated herpesvirus(KSHV; also known as human herpesvirus 8(HHV-8)), and its associated diseases. Here we review the current knowledge of KSHV biology and pathogenesis, with a particular emphasis on new and exciting advances in the field of epigenetics. We also discuss the development and practicality of various cell culture and animal model systems to study KSHV replication and pathogenesis.     

A Unique Protease Cleavage Site Predicted in the Spike Protein of the Novel Pneumonia Coronavirus (2019-nCoV) Potentially Related to Viral Transmissibility

正Dear Editor,In December 2019, a novel human coronavirus caused an epidemic of severe pneumonia(Coronavirus Disease 2019,COVID-19) in Wuhan, Hubei, China(Wu et al. 2020; Zhu et al. 2020). So far, this virus has spread to all areas of China and even to other countries. The epidemic has caused 67,102 confirmed infections with 1526 fatal cases     

Curcuminoids as inhibitors of thioredoxin reductase: A receptor based pharmacophore study with distance mapping of the active site

Curcumin is the yellow pigment of turmeric that interacts irreversibly forming an adduct with thioredoxin reductase (TrxR), an enzyme responsible for redox control of cell and defence against oxidative stress. Docking at both the active sites of TrxR was performed to compare the potency of three naturally occurring curcuminoids, namely curcumin, demethoxy curcumin and bis-demethoxy curcumin. Results show that active sites of TrxR occur at the junction of E and F chains. Volume and area of both cavities is predicted. It has been concluded by distance mapping of the most active conformations that Se atom of catalytic residue SeCYS498, is at a distance of 3.56 from C13 of demethoxy curcumin at the E chain active site, whereas C13 carbon atom forms adduct with Se atom of SeCys 498. We report that at least one methoxy group in curcuminoids is necessary for interation with catalytic residues of thioredoxin. Pharmacophore of both active sites of the TrxR receptor for curcumin and demethoxy curcumin molecules has been drawn and proposed for design and synthesis of most probable potent antiproliferative synthetic drugs.     

The structure, reproduction and taxonomy of Vidalia and Osmundaria (Rhodophyta, Rhodomelaceae)

Comprises species occurring mostly in subtidal habitats in tropical, subtropical and warm-temperate areas of the world. An analysis of the type species, V. spiralis (Sonder) Lamouroux ex J. Agardh, a species from Australia, establishes basic characters for distinguishing species in the genus. These characters are (1) branching patterns of thalli, (2) flat blades that may be spiralled on their axis, (3) width of the blade, (4) primary or secondary derivation of sterile and fertile branchlets and (5) position of sterile and fertile branchlets on the thalli. Application of the latter two characters provides an important basic method for separation of species into three major groups. Osmundaria , a genus known only in southern Australia, was studied in relation to Vidalia , and its separation from the Vidalia assemblage is not accepted. Species of Vidalia therefore are transferred to the older genus name, Osmundaria. Two new species, Osmundaria papenfussii and Osmundaria oliveae are described from Natal. Confusion in the usage of the epithet, Vidalia fimbriala Brown ex Turner has been clarified, and Vidalia gregaria Falkenberg, described as an epiphyte on Osmundaria pro/ifera Lamouroux, is revealed to be young branches of the host, Osmundaria prolifera.     

New or rare chromosome counts in Artemisia L. (Asteraceae, Anthemideae) and related genera from Kazakhstan

Fifteen chromosome counts of six Artemisia taxa and one species of each of the genera Brachanthemum, Hippolytia, Kaschgaria, Lepidolopsis and Turaniphytum are reported from Kazakhstan. Three of them are new reports, two are not consistent with previous counts and the remainder are confirmations of very scarce (one to four) earlier records. All the populations studied have the same basic chromosome number, x = 9, with ploidy levels ranging from 2x to 6x. Some correlations between ploidy level, morphological characters and distribution are noted.