泵抑制剂对嗜麦芽寡养单胞菌临床分离株氟喹诺酮类敏感性的影响

目的 了解嗜麦芽寡养单胞菌临床分离株外排泵的分布以及不同泵抑制剂对该菌氟喹诺酮类敏感性的影响。方法 用琼脂二倍稀释法检测泵抑制剂存在时，嗜麦芽寡养单胞菌对环丙沙星、氧氟沙星和诺氟沙星的敏感性变化。结果 羰基氰氯苯腙(CCCP)和利舍平降低部分嗜麦芽寡养单胞菌株对氟喹诺酮的耐药性。维拉帕米无明显泵抑制作用。泵阳性株存在于耐药和非耐药菌株中，集中于耐药性较高的菌株。泵抑制剂对该菌外排氧氟沙星的抑制大于对诺氟沙星和环丙沙星的作用。结论 多重外排泵是嗜麦芽寡养单胞菌对氟喹诺酮类耐药的原因之一。泵抑制剂可部分逆转这种耐药性。     

氟喹诺酮类药物对嗜麦芽寡养单胞菌的抗菌活性及羰基氰氯苯腙对其的影响

目的　了解氟喹诺酮类药物 (FQNs)对嗜麦芽寡养单胞菌的抗菌活性及羰基羟基氰氯苯腙(CCCP)对其抗菌活性的影响。方法　采用琼脂二倍稀释法测定抗菌药物的最低抑菌浓度 (MIC) ,同时测定CCCP对 FQNs的 MIC值的影响。结果　 14 4株嗜麦芽寡养单胞菌对多种常用抗生素呈现多重耐药 ,但对SMZ/ TMP、替卡西林 /克拉维酸以及 FQNs的耐药率较低 ,尤其是新型 FQNs具有很高的抗菌活性 ,其中抗菌活性由高到低依次是加替沙星、司帕沙星、左氧氟沙星和环丙沙星。主动外排泵抑制剂 CCCP在体外能增强 FQNs的抗菌活性 ,主动外排机制不仅存在于 FQNs耐药菌株 ,而且也存在于 FQNs敏感菌株 ,但是CCCP对 4种 FQNs耐药菌株的影响更大。结论　嗜麦芽寡养单胞菌的感染经验上可选用 SMZ/ TMP或 FQNs治疗 ,对于严重感染可采用 SMZ/ TMP和替卡西林 /克拉维酸联合或用新型 FQNs治疗。主动外排泵抑制剂在体外的确能降低 FQNs对嗜麦芽寡养单胞菌的 MIC,若能保证安全性 ,可望用于临床克服细菌的耐药性。     

嗜麦芽寡养单胞菌临床株外排泵SmeDEF诱导表达的研究

目的了解嗜麦芽寡养单胞菌的临床株H407和A26在抗菌药物诱导下耐药性和SmeDEF外排泵表达的变化。方法比较有或无环丙沙星诱导时嗜麦芽寡养单胞菌H407和A26的MIC和EOP值,对在亚抑菌浓度环丙沙星中培育后的菌株smeD基因进行RT-PCR扩增,监测表达水平。结果对抗生素敏感的H407和A26泵抑制呈阳性或阴性反应。2株菌经环丙沙星过夜孵育后测得的对氯霉素和环丙沙星MIC值比正常非诱导时的MIC值高1~3个稀释度;加泵抑制剂CCCP后,2个诱导株的MIC值均有明显下降。H407和A26经环丙沙星诱导后氯霉素的EOP分别有4.8和近4倍的增长。2株菌在抗菌药物过夜和指数期添加生长时smeD的mRNA明显高于无抗菌药物培养株。结论2株嗜麦芽寡养单胞菌临床株在环丙沙星诱导下对部分抗菌药物的耐药性提高,与SmeDEF泵的诱导表达增加有关。     

氟喹诺酮类药物对嗜麦芽窄食单胞菌主动外排泵表型特征研究

目的 研究氟喹诺酮类药物(FQNS)对嗜麦芽窄食单胞菌的抗菌活性及氰氯苯腙(CCCP)对其主动外排泵表型特征的影响。方法　用琼脂二倍稀释法测定FQNS的最低抑菌浓度(MIC),测定CCCP对FQNS的MIC值的影响。结果　111株嗜麦芽窄食单胞菌对FQNS耐药率较低,尤以新型FQNS抗菌活性较高,其中抗菌活性由高到低依次是加替沙星、司帕沙星、左氧氟沙星和环丙沙星。主动外排泵抑制剂CCCP在体外能增强FQNS抗菌活性,主动外排机制即存在于FQNS耐药菌株,也存在于FQNS敏感菌株。但是CCCP对4种FQNS耐药菌株影响更大。结论　主动外排泵抑制剂在体外能降低FQNS对嗜麦芽窄食单胞菌的MIC,若能保证安全性,可能成为临床克服细菌耐药性的一种方法。     

4种氟喹诺酮类药物对嗜麦芽寡养单胞菌防耐药变异浓度的测定

目的为了解4种氟喹诺酮类药物对嗜麦芽寡养单胞菌的防耐药变异能力,以指导临床合理用药。方法采用标准琼脂二倍稀释法测定莫西沙星、左氧氟沙星、洛美沙星、环丙沙星对嗜麦芽寡养单胞菌的最低抑菌浓度;将总量为1.2×1010CFU的细菌接种于含不同浓度药物的琼脂平皿上,无菌落生长的最低药物浓度即为该药的防耐药变异浓度(MPC)。结果莫西沙星和左氧氟沙星对嗜麦芽寡养单胞菌的敏感率为100%,洛美沙星敏感率为64.5%,环丙沙星敏感率为9.7%。莫西沙星和左氧氟沙星MPC90为64mg/L,洛美沙星和环丙沙星MPC90为256m g L/。4种药物的M PC90/MIC90均为64。结论莫西沙星和左氧氟沙星对嗜麦芽寡养单胞菌具有高度的抗菌活性,但结合药代动力学参数,4种药物血药浓度均位于MIC和MPC之间,提示单药治疗易导致耐药突变菌株的富集生长,应联合用药以限制细菌耐药的发生。     

嗜麦芽寡养单胞菌感染及其耐药性分析

目的：了解嗜麦芽寡养单胞菌感染的临床及耐药性特点。方法：1997年7月－2000年6月从临床送检标本分离114株嗜麦芽寡养单胞菌，对其进行抗菌药物敏感试验和诱导型β－内酰胺酶测定。结果：114例嗜麦芽寡养单胞菌感染病例中，肺部感染为58．77％，泌尿系感染为25．44％。该菌对氨苄西林、羧苄西林、头孢唑林、头孢呋辛、头孢曲松、头孢他啶、庆大霉素、阿米卡星的耐药率均在60％以上，对亚胺培南的耐药率几乎为100％，但对替卡西林／克拉维酸、环丙沙星、SMZ／TMP、环丙沙星或替卡西林／克拉维酸。     

氟喹诺酮结构及药敏测定方法对嗜麦芽寡养单胞菌敏感性的影响

目的探讨不同结构氟喹诺酮及不同药敏测定方法对嗜麦芽寡养单胞菌敏感性的影响，以指导临床合理选择抗菌药物。方法采用标准琼脂二倍稀释法测定5种氟喹诺酮对92株嗜麦芽寡养单胞菌临床分离株及20株实验室环丙沙星耐药株的最低抑菌浓度（MIC）；采用Kirby Bauer纸片扩散法测定环丙沙星对92株嗜麦芽寡养单胞菌的敏感性。结果莫西沙星、加替沙星、左氧氟沙星、洛美沙星和环丙沙星对临床分离株的敏感率分别为94．6％、90．2％、87．0％、66．3％和32．6％；莫西沙星、加替沙星、左氧氟沙星和洛美沙星对实验室耐药株的敏感率分别为75％、60％、60％和25％。与琼脂法比较，纸片法测定的环丙沙星敏感率增加、耐药率降低，均有统计学差异（P〈0．01，n=92）。结论不同结构的氟喹诺酮对嗜麦芽寡养单胞菌的敏感性不同，环丙沙星敏感性不能代替所有氟喹诺酮的敏感性；纸片法测定的敏感性不能代替琼脂法测定的敏感性。     

志贺菌属外排泵AcrAB-TolC及其调控基因突变与氟喹诺酮耐药性的关系

目的 探讨临床分离耐氟喹诺酮志贺菌外排泵Acr AB-Tol C基因及其调控基因mar OR、acr R、sox S突变与耐药性的关系。方法 使用K-B纸片扩散法筛选临床耐药菌,测定加入泵抑制剂羰基氢氯苯腙(CCCP)后萘啶酸、左氧氟沙星、氧氟沙星、环丙沙星、诺氟沙星的最低抑菌浓度(MIC)的变化,PCR扩增外排泵基因acr A、acr B及其调控基因mar OR、acr R、sox S并测序。结果 159株志贺菌中共筛选出11株氟喹诺酮耐药菌株;加入质子泵抑制剂后2株耐药菌株对氟喹诺酮类抗菌药的MIC下降;7株耐药菌对氟喹诺酮类抗菌药的MIC不变;2株耐药菌对氟喹诺酮类抗菌药的MIC上升。基因测序发现氟喹诺酮耐药菌株外排泵Acr AB-Tol C调控基因mar OR第36、37、38、39位存在CATT碱基缺失。结论 泵抑制剂可部分抑制外排泵的活性。mar OR突变可能与志贺菌对氟喹诺酮类抗菌药耐药有关。     

2株嗜麦芽寡养单胞菌对β-内酰胺类耐药性的传递

目的了解嗜麦芽寡养单胞菌临床株对抗生素的耐药情况及耐药性的可传递性。方法琼脂纸片扩散法药物敏感试验检测嗜麦芽寡养单胞菌的抗生素耐药情况。接合传递实验获耐药接合株。结果临床菌株H417传递的抗生素耐药性是对头孢噻肟、头孢哌酮、羧苄西林、头孢他啶及庆大霉素；临床菌株A3573传递的耐药性有头孢噻肟、头孢他啶、羧苄西林、氨苄西林／克拉维酸和氨曲南。同时伴有质粒的传递。结论嗜麦芽寡养单胞菌临床株对β-内酰胺类及氨基糖苷类的耐药性可经质粒介导传递：     

嗜麦芽寡养单胞菌对黏菌素的耐药特征及体外联合抗菌活性研究

目的 研究黏菌素分别与氯霉素、左氧氟沙星、头孢他啶、米诺环素及复方磺胺甲噁唑联用对多重耐药嗜麦芽寡养单胞菌的体外抗菌活性。方法 收集2015—2018年分离自温州医科大学附属第一医院的431株嗜麦芽寡养单胞菌;采用微量肉汤稀释法检测黏菌素对嗜麦芽寡养单胞菌的最低抑菌浓度(minimal inhibitory concentration, MIC),计算逐年耐药率;通过棋盘法和微量肉汤稀释法检测黏菌素分别与氯霉素、左氧氟沙星、头孢他啶、米诺环素与复方磺胺甲噁唑联用及各自单用时对多重耐药嗜麦芽寡养单胞菌的MIC值,并通过计算部分抑菌浓度指数(FICI)评价联合抑菌效果。结果 2015—2018年间,我院嗜麦芽寡养单胞菌对黏菌素的耐药率呈现逐年上升趋势;黏菌素与氯霉素、左氧氟沙星及米诺环素联用后均表现为协同或相加作用;与头孢他啶及复方磺胺甲噁唑联用都存在无关作用,未发现拮抗作用。结论 黏菌素与氯霉素、左氧氟沙星、米诺环素联合对多重耐药嗜麦芽寡养单胞菌具有较好的体外抗菌活性,将对临床联合用药治疗多重耐药嗜麦芽寡养单胞菌的选择更具指导意义。     

Efficacy of gut lavage,hemodialysis, and hemoperfusion in the therapy of paraquat or diquat intoxication

Clinical and in vitro investigations were carried out to test the efficacy of gut lavage, hemodialysis, and hemoperfusion in the treatment of poisoning with paraquat or diquat. In a patient suffering from diquat intoxication 130 times more diquat was removed by gut lavage 30 h after ingestion than was removed by complete aspiration of the gastric contents.Determination of in vitro clearances for paraquat and diquat by hemodialysis showed that, at serum concentrations of 1–2 ppm, such as are frequently encountered in poisoning in man, toxicologically relevant quantities of herbicide cannot be removed from the body. At a concentration of 20 ppm, on the other hand, hemodialysis proved to be effective, the clearance being 70 ml/min at a blood flow rate of 100 ml/min. The efficacy of hemoperfusion with coated activated charcoal was on the whole better. Especially at concentrations around 1–2 ppm, the clearance values for hemoperfusion were some 5–7 times higher than those for hemodialysis.In a patient suffering from paraquat poisoning, both hemodialysis as well as hemoperfusion were carried out. The in vitro results could be confirmed: At serum concentrations of paraquat less than 1 ppm no clearance could be obtained by hemodialysis while by hemoperfusion with activated charcoal quite high clearance values were measured and the serum level dropped down to zero.

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Zusammenfassung Klinische Untersuchungen und Laboratoriumsversuche wurden durchgeführt, um die Wirksamkeit von Darmspülung, Hämodialyse und Hämoperfusion bei Paraquat- und Deiquat-Vergiftungen zu prüfen.Bei einem Patienten wurde 30 Std nach Deiquat-Aufnahme durch Darmspülung 130mal mehr Deiquat entfernt als durch vollständige Aspiration des Mageninhaltes. In vitro-Versuche ergaben, daß bei Blutserumkonzentrationen von 1–2 ppm, die bei Vergiftungen oft gemessen werden, durch Hämodialyse keine toxikologisch relevanten Paraquat- oder Deiquat-Mengen entfernt werden können. Dagegen erwies sich die Hämodialyse bei 20 ppm und einer Blutumlaufgeschwindigkeit von 100 ml/min mit einer Clearance von 70 ml/min als wirksam. Die Hämoperfusion mit beschicheter Aktivkohle war in diesen Versuchen aber eindeutig überlegen, denn insbesondere bei Konzentrationen um 1–2 ppm waren die Clearance-Werte 5–7mal höher als bei der Hämodialyse.Die in vitro-Ergebnisse wurden bei einem Patienten mit einer Paraquat-Vergiftung bestätigt: Bei Konzentrationen unter 1 ppm war die Hämodialyse wirkungslos, während durch Hämoperfusion relativ hohe Clearance-Werte erreicht wurden, so daß der Serumspiegel rasch unter die Nachweisgrenze abfiel.

     

In vitro studies on sterically stabilized liposomes (SL) as enzyme carriers in organophosphorus (OP) antagonism

This study describes a new approach for organophosphorous (OP) antidotal treatment by encapsulating an OP hydrolyzing enzyme, OPA anhydrolase (OPAA), within sterically stabilized liposomes. The recombinant OPAA enzyme was derived from Alteromonas strain JD6. It has broad substrate specificity to a wide range of OP compounds: DFP and the nerve agents, soman and sarin. Liposomes encapsulating OPAA (SL)* were made by mechanical dispersion method. Hydrolysis of DFP by (SL)* was measured by following an increase of fluoride ion concentration using a fluoride ion selective electrode. OPAA entrapped in the carrier liposomes rapidly hydrolyze DFP, with the rate of DFP hydrolysis directly proportional to the amount of (SL)* added to the solution. Liposomal carriers containing no enzyme did not hydrolyze DFP. The reaction was linear and the rate of hydrolysis was first order in the substrate. This enzyme carrier system serves as a biodegradable protective environment for the recombinant OP-metabolizing enzyme, OPAA, resulting in prolongation of enzymatic concentration in the body. These studies suggest that the protection of OP intoxication can be strikingly enhanced by adding OPAA encapsulated within (SL)* to pralidoxime and atropine.     

Aquatic Insects and Trace Metals: Bioavailability,Bioaccumulation, and Toxicity

Abstract

The uptake of metals from food and water sources by insects is thought to be additive. For a given metal, the proportions taken up from water and food will depend both on the bioavailable concentration of the metal associated with each source and the mechanism and rate by which the metal enters the insect. Attempts to correlate insect trace metal concentrations with the trophic level of insects should be made with a knowledge of the feeding relationships of the individual taxa concerned. Pathways for the uptake of essential metals, such as copper and zinc, exist at the cellular level, and other nonessential metals, such as cadmium, also appear to enter via these routes. Within cells, trace metals can be bound to proteins or stored in granules. The internal distribution of metals among body tissues is very heterogeneous, and distribution patterns tend to be both metal and taxon specific. Trace metals associated with insects can be both bound on the surface of their chitinous exoskeleton and incorporated into body tissues. The quantities of trace meals accumulated by an individual reflect the net balance between the rate of metal influx from both dissolved and particulate sources and the rate of metal efflux from the organism. The toxicity of metals has been demonstrated at all levels of biological organization: cell, tissue, individual, population, and community. Much of the literature pertaining to the toxic effects of metals on aquatic insects is based on laboratory observations and, as such, it is difficult to extrapolate the data to insects in nature. The few experimental studies in nature suggest that trace metal contaminants can affect both the distribution and the abundance of aquatic insects. Insects have a largely unexploited potential as biomonitors of metal contamination in nature. A better understanding of the physico-chemical and biological mechanisms mediating trace metal bioavailability and exchange will facilitate the development of general predictive models relating trace metal concentrations in insects to those in their environment. Such models will facilitate the use of insects as contaminant biomonitors.     

Alzheimer’s disease – current trends in basic and clinical research. Highlights from the 16th ECNP

Advances in the molecular biological knowledge of neuronal nicotinic acetylcholine receptors (nAChRs) have led to a growing interest by the pharmaceutical industry in the development of novel compounds that selectively modulate nAChR function. The ability of (-)-nicotine, an activator of nAChRs, to enhance attentional aspects of cognition in animals and humans, to exert neuroprotective and anxiolytic-like effects, and presumably to mediate the negative correlation between smoking and Alzheimer's (and Parkinson's) Disease, has focused interest on the potential therapeutic utility of modulators of nAChR function for treatment of some of the deficits associated with these progressive, neurodegenerative conditions. Numerous compounds are known which activate nAChRs and which might serve as lead compounds toward the development of such agents. The pharmacologic diversity of neuronal nAChR subtypes suggests the possibility of developing selective compounds which would have more favourable side-effect profiles than existing agents. This broader class of agents, collectively called cholinergic channel modulators (ChCMs), is anticipated to encompass compounds which would have more favourable side-effect profiles than existing agents, which generally exhibit low selectivity. This selectivity may be achieved by preferentially activating some subtypes of nAChRs (i.e., Cholinergic Channel Activators, ChCAs) or inhibiting the function of other subtypes (Cholinergic Channel Inhibitors, ChCIs). An overview of the biology of nAChRs and the rationale for the use of ChCMs for the treatment of dementia related to neurodegenerative diseases are presented, followed by a discussion of lead compounds and compounds under consideration for clinical evaluation.     

Steroidal sapogenin contents in some domestic plants

In order to find out the values of the steroid resources for the future use. the compositions and contents of steroidal sapogenins from 13 domestic plants have been investigated. As a result,Dioscorea nipponica, D. quinqueloba andSmilax china were found to have large amount of diosgenin. And pennogenin inTrillium kamtschaticum andParis verticillata, yuccagenin inAllium fistulosum, hecogenin inAgave americana and neochlorogenin inSolanum nigum were appeared to be major steroidal sapogenins.