鲍曼不动杆菌菌毛在细菌生物膜形成中的作用

目的通过观察鲍曼不动杆菌菌毛,了解菌毛结构在生物被膜形成过程中的作用。方法以ICU的医院感染患者的腹腔手术后引流液、痰及呼吸机导管内壁附着物等为材料分离鉴定细菌,制备细菌的电镜标本,通过超微结构观察鲍曼不动杆菌菌体表面的菌毛与生物被膜形成的相关性。结果新分离的鲍曼不动杆菌菌体表面存在菌毛,菌毛与生物膜形成过程中的粘附有关。结论菌毛粘附是生物被膜形成的原因之一。     

乳酸锌和氟化亚锡对铜绿假单胞菌、鲍曼不动杆菌和变异链球菌生物被膜的抑制作用

乳酸锌(Zn lactate·3H_2O)和氟化亚锡(SnF_2)常作为牙膏中的活性物质添加剂用来预防龋齿及口腔生物被膜的形成。文中评估了Zn lactate·3H_2O和SnF_2对铜绿假单胞菌、鲍曼不动杆菌和变异链球菌生物被膜的作用。对铜绿假单胞菌PAO1生物被膜的抑制实验证实乳酸锌和氟化亚锡都具有抑制其生物被膜的功能,联用效果尤佳。乳酸锌通过干扰胞外多糖基质网的形成起作用,而氟化亚锡则可以明显降低生物被膜的生物量。更为重要的是,工作浓度的两种化合物联用几乎可以完全抑制3种实验菌株生物被膜的形成。     

黄酮化合物对耐药性鲍曼不动杆菌抑菌作用研究

为探索中药黄酮化合物对耐药性鲍曼不动杆菌的抑菌效果。研究通过微量肉汤稀释法,考察40种中药及天然来源的黄酮化合物对三株耐药性鲍曼不动杆菌菌株的体外抑菌作用,测试其最低抑菌浓度与最低杀菌浓度;采用结晶紫染色法进一步测定目标化合物对鲍曼不动杆菌生物被膜形成的能力;探索黄酮联合头孢哌酮/舒巴坦对鲍曼不动杆菌的抑菌效果。研究表明,黄酮单体对于鲍曼不动杆菌具有不同程度的体外抑菌作用。其中部分具有轻微作用,其最小抑菌浓度仅为0.5 mg/mL;黄芩素和汉黄芩苷具有显著的抑菌作用,其最小抑菌浓度分别为0.062 5和0.125 mg/mL;同时,黄芩素对鲍曼不动杆菌生物被膜形成具有明显的抑制作用。此外,黄芩素和头孢哌酮/舒巴坦联合作用于耐药鲍曼不动杆菌与单用黄芩素或头孢哌酮/舒巴坦比较,浓度有所降低。综上,黄芩素对耐药性鲍曼不动杆菌有较好的体外抑菌效果,这为治疗耐药性鲍曼不动杆菌引发的感染的治疗提供新思路和方向。     

鲍曼不动杆菌临床菌株生物膜形成能力的研究

目的对鲍曼不动杆菌临床菌株的生物膜形成能力进行对比研究,并分析生物膜形成的一些可能的影响因素。方法利用在聚苯乙烯板上构建生物膜的技术对51株全耐药的鲍曼不动杆菌的生物膜形成能力进行检测,同时对分离自下呼吸道和无菌体液的各20株鲍曼不动杆菌的生物膜形成能力进行研究,然后对新近报道的鲍曼不动杆菌生物膜形成相关基因abaI在所有菌株中的分布情况进行检测。结果 51株全耐药的鲍曼不动杆菌中35株(68.6%)可以形成生物膜,并且形成生物膜的能力较强。50%(10/20)分离自下呼吸道的鲍曼不动杆菌能够形成生物膜,20株无菌体液中分离的菌株仅有1株可以形成生物膜。78.0%(71/91)鲍曼不动杆菌中abaI基因扩增阳性。结论分离自下呼吸道的鲍曼不动杆菌临床菌株有较强的生物膜形成能力,abaI基因广泛存在于鲍曼不动杆菌临床菌株中。临床在治疗鲍曼不动杆菌感染的同时需要考虑其在感染部位形成生物膜的因素,可能在治疗的同时有必要加入一些对生物膜有穿透性的药物。     

鲍曼不动杆菌耐药持留菌的特征及Ⅱ型毒素-抗毒素系统的多样性

摘要:【目的】研究鲍曼不动杆菌持留菌的特征,分析鲍曼不动杆菌Ⅱ型毒素-抗毒素(TA)系统的多样性及分布,探讨TA系统与持留菌形成的潜在关联。【方法】采用分属于6 大类中的6种不同抗生素分别筛选的方法分离并定量鲍曼不动杆菌的持留菌;应用PSI-BLAST及TBLASTN程序分析721株鲍曼不动杆菌中的Ⅱ型TA系统;通过聚合酶链式反应(PCR)方法检测44株临床多重耐药(MDR)的鲍曼不动杆菌中5个Ⅱ型TA系统的分布。【结果】我们发现不同抗生素处理得到的鲍曼不动杆菌持留菌的数量并不相同;对多数抗生素而言,浓度越高,得到的持留菌数量越少;处于生长平台期的菌株中相比对数期含有更多的持留菌;多粘菌素B及妥布霉素均具有杀灭部分持留菌的能力;鲍曼不动杆菌所有菌株中均含有Ⅱ型TA系统,HTH/GNAT类型在菌株中分布最为广泛;所有700余株鲍曼不动杆菌基因组中共含有15个潜在的Ⅱ型TA系统;三类普遍存在于已知基因组中的明确功能的Ⅱ型TA系统在临床多重耐药菌株中同样分布广泛,且HigB/HigA的表达量在标准株的持留菌中显著增高。【结论】鲍曼不动杆菌的持留菌水平与菌株的生长状态、抗生素的种类及浓度密切相关。Ⅱ型TA系统在鲍曼不动杆菌已知基因组和临床多重耐药菌株中普遍存在,其中,HTH/GNAT、GP49/Cro(HigB/HigA)及DUF497/COG3514三种类型的Ⅱ型TA系统可能在鲍曼不动杆菌持留菌的形成中扮演重要角色。     

鲍曼不动杆菌的耐药机制研究进展

鲍曼不动杆菌是临床常见感染菌,耐药性日益增强。其多重耐药性与可长期存活性将导致菌膜的形成,而这种可以抵抗抗菌素治疗的菌膜有着复杂的机构,在复杂结构中起到形成菌膜、维持菌膜稳定性的两个重要组成部分为胞外多糖(EPS)和菌膜相关性蛋白(the biofilm-associated protein,Bap),促使鲍曼不动杆菌躲避宿主免疫系统的攻击,因此针对鲍曼不动杆菌的治疗也愈发困难。     

铜绿假单胞菌体外抑菌活性研究

目的 观察铜绿假单胞菌抗菌物质对鲍曼不动杆菌等细菌的体外抑菌效果.方法 用交叉条带实验方法检测了铜绿假单胞菌对鲍曼不动杆菌、耐甲氧西林表皮葡萄球菌和粪肠球菌的体外抑制活性.结果 铜绿假单胞菌对鲍曼不动杆菌、耐甲氧西林表皮葡萄球菌和粪肠球菌体外抑菌活性良好,10株铜绿假单胞菌中,有8株对鲍曼不动杆菌的抑制率均达到了100％.另外有8株对耐甲氧西林表皮葡萄球菌的抑菌率均为100％;有6株对粪肠球菌的抑菌率为100％.结论 铜绿假单胞菌对上述3种致病菌具有较强的抗菌活性,具有开发前景.     

鲍曼不动杆菌裂解性噬菌体LZ35的 分离及全基因组分析

摘要：目的 通过分离鉴定鲍曼不动杆菌噬菌体并进行遗传信息分析,为今后噬菌体用于治疗鲍曼不动杆菌引起的感染提供依据。方法 以鲍曼不动杆菌临床分离株为宿主菌,从医院污水中分离鲍曼不动杆菌噬菌体并进行纯化、电镜观察形态特征、提取噬菌体DNA,进行全基因组测序,分析全基因组的结构特征,比较基因组分析其进化关系。结果 分离到鲍曼不动杆菌裂解性噬菌体LZ35,电镜观察显示,该噬菌体属于有尾噬菌体目肌尾病毒科。基因组全长44 885 bp,G＋C含量为37.95%,含有83个开放阅读框,其中22个编码序列可预测其功能,61个编码序列为未知基因。噬菌体LZ35的基因组与鲍曼不动杆菌噬菌体IME-AB2和YMC-13-01-C62具有很高的同源性（分别为97%和99%）,与鲍曼不动杆菌噬菌体YMC11/12/R1215的进化关系最近。结论 以鲍曼不动杆菌临床分离株为宿主菌,分离到鲍曼不动杆菌裂解性噬菌体LZ35,明确了其形态和基因组特征,为防治噬菌体疗法奠定基础。     

重症监护病房鲍曼不动杆菌和铜绿假单胞菌的分布及耐药性监测

目的探讨医院重症监护病房(ICU)鲍曼不动杆菌及铜绿假单胞菌临床分离株的分布及耐药性特点。方法回顾性调查2006年7月至2010年6月ICU患者中分离出的鲍曼不动杆菌和铜绿假单胞菌的临床感染情况,对111株鲍曼不动杆菌和73株铜绿假单胞菌的药敏结果进行统计分析,所有数据采用WHONET 5.5软件进行分析。结果鲍曼不动杆菌及铜绿假单胞菌在呼吸道标本中检出率最高(76.1%),其次是CVP导管尖端(8.7%),鲍曼不动杆菌对临床常用抗菌药物耐药率85%的有8种,对亚胺培南的耐药率也达70.3%,耐药率最低的仅有头孢哌酮/舒巴坦(17.1%);铜绿假单胞菌对替卡西林、亚胺培南有较高的耐药率,分别为45.2%、41.1%,对其他抗菌药物耐药率均25%。结论下呼吸道是ICU患者鲍曼不动杆菌和铜绿假单胞菌感染的主要部位;ICU患者感染以鲍曼不动杆菌和铜绿假单胞菌为主,鲍曼不动杆菌较铜绿假单胞菌耐药及多重耐药性严重。     

从耐药鲍曼不动杆菌中发现β-内酰胺酶ADC基因和膜孔蛋白carO基因新的变异型

目的 调查一组耐药鲍曼不动杆菌中β-内酰胺酶基因和膜孔蛋白基因的存在和变异情况.方法 收集2010年1月至2010年12月浙江某医院ICU患者痰液标本中分离的多耐药和泛耐药鲍曼不动杆菌各10株,用分子鉴定法鉴定菌种,再用聚合酶链反应(PCR)及序列分析的方法分析34种β-内酰胺酶基因与膜孔蛋白carO基因.结果 本组20株耐药鲍曼不动杆菌共检出TEM-1型20株(100％)、OXA-23型10株(50.0％)、ADC-30型12株(60.0％)、ADC基因新的变异型ADC-60型8株(40.0％)(GenBank登录号:JQ692087).10株PDR菌均检出OXA-23型β-内酰胺酶基因,而10株MDR菌则均未检出OXA-23型β-内酰胺酶基因.20株耐药鲍曼不动杆菌膜孔蛋白carO基因均存在有义突变,19株测得序列相同,翻译成氨基酸序列后与鲍曼不动杆菌敏感株(SDF)比较,一致率为76.0％.8号株carO基因序列与其他19株测得DNA序列不同,第351位缺失了12个碱基并导致过早出现终止密码子.结论 本组鲍曼不动杆菌对β-内酰胺类药物耐药主要与产TEM、ADC、OXA-23和carO基因存在有义突变相关.OXA-23型β-内酰胺酶基因阳性是PDR菌耐碳青霉烯类药物的原因.ADC基因存在新变异型:ADC-60是国内外首次报道.     

嗜水气单胞菌生物被膜对其耐药性的影响

建立了嗜水气单胞菌(Aeromonas hydrophila, Ah)的生物被膜(Bacterial biofilm, BF) 体外形成模型, 并对11种抗菌药物对BF细菌和浮游（Freecell, FC）细菌的清除作用进行了研究。将Ah J1株在放有硅胶膜的TSB中培养7d,用银染法鉴定,发现可形成良好的BF。FC细菌对青霉素具有耐药性, 最低杀菌浓度(MBC)为256μg/mL;对蒽诺沙星和氟哌酸最敏感,MBC分别为003μg/mL和0.25μg/mL。氟苯尼考对BF细菌的清除能力最强,作用于BF细菌和FC细菌的MBC之比为2∶1;卡那霉素、青霉素、新霉素的MBC比值在32∶1以上。扫描电镜观察蒽诺沙星作用于FC及BF细菌前后的形态变化,并测定其杀菌曲线。发现4×MBC时可完全清除FC细菌,但不能完全清除BF细菌;在32×MBC时,4h内可完全清除FC细菌,而24h内完全清除BF细菌。结果表明形成BF的Ah对抗菌药物可形成强耐受性,其潜在影响应引起足够重视。     

High dietary aluminum affects the response of rats to silicon deprivation

Antagonistic interactions between silicon and aluminum occur in living organisms. Thus, an experiment was performed to ascertain whether high dietary aluminum would accentuate the signs of silicon deprivation in rats and conversely whether silicon deprivation would accentuate the response to high dietary aluminum. The experiment was factorially arranged with two variables: silicon as sodium metasilicate, 0 or 40 μg/g diet, and aluminum as aluminum citrate, 0 or 500 μg/g diet. After 9 wk, body weights and plasma urea nitrogen were higher and plasma concentrations of threonine, serine, glycine, cystine, and methionine were lower in silicon-adequate than silicon-deprived rats. High dietary aluminum significantly decreased plasma phenylalanine. An interaction between aluminum and silicon affected plasma triglyceride, cholesterol, and phosphorus concentrations. High dietary aluminum decreased these variables when silicon was absent from the diet, but increased them when silicon was present. Skull iron and silicon concentrations were decreased and iron and zinc concentrations in the femur were increased by the addition of 500 μg Al/g diet. High dietary aluminum decreased tibia density in silicon-adequate rats, but increased tibial density in silicon-deprived rats. The findings indicate that in rats, high dietary aluminum can affect the response to silicon deprivation and dietary silicon can affect the response to high dietary aluminum.     

Prior exercise training increases collateral-dependent blood flow in rats after acute femoral artery occlusion

We evaluated whether prior training would improve collateral blood flow (BF) to the calf muscles after acute-onset occlusion of the femoral artery. Exercise training was performed in the absence of any vascular occlusion. Adult male Sprague-Dawley rats ( approximately 325 g) were kept sedentary (n = 14), limited to cage activity, or exercise trained (n = 14) for 6 wk by treadmill running. Early in the day of measurement, animals were surgically prepared for BF determination, and the femoral arteries were occluded bilaterally. Four to five hours later, collateral BF was determined twice during treadmill running with the use of (141)Ce and (85)Sr microspheres: first, at a demanding speed and, second, after a brief rest and at a higher speed. The absence of any further increase in BF at the higher speed indicated that maximal collateral BF was measured. Prior training increased calf muscle BF by approximately 70% compared with sedentary animals; however, absolute BF remained below values previously observed in animals with a well-developed collateral vascular tree. Thus prior training appeared to optimize the use of the existing collateral circuit. This implies that altered vasoresponsiveness induced in normal nonoccluded vessels with exercise training serves to improve collateral BF to the periphery.     

一株洛菲不动杆菌对碳青霉烯类抗生素耐药机制的研究

目的研究洛菲不动杆菌对亚胺培南、美洛培南耐药的分子机制。方法K-B纸片琼脂扩散法检测洛菲不动杆菌B69对头孢他啶、头孢曲松、环丙沙星、阿米卡星的耐药性,琼脂对倍稀释法检测B69对亚胺培南、美洛培南的最低抑菌浓度;PCR扩增OXA、IMP、VIM型碳青霉烯酶基因,测序确定耐药基因型别;粗提酶水解亚胺培南纸片试验检测酶活性。结果洛菲不动杆菌B69具有多重耐药性;PCR扩增IMP基因阳性,经测序为IMP-8;粗提酶水解亚胺培南。结论产IMP-8型金属β-内酰胺酶是洛菲不动杆菌B69对碳青霉烯耐药的重要机制。     

鲍曼不动杆菌超广谱β-内酰胺酶检测及耐药性分析

为了解厦门中山医院鲍曼不动杆菌产超广谱β 内酰胺酶(ESBLs)情况及耐药现状,指导临床用药,采用美国临床实验室标准化委员会(NCCLS)推荐的纸片扩散法,测定 2002 ~2003年临床分离的 211株鲍曼不动杆菌产ESBLs的比例及其对头孢噻肟、阿米卡星等 13种抗菌药物耐药性特征。结果表明 211株鲍曼不动杆菌中ESBLs阳性 27株(占 12. 8% ),产ESBLs菌株对 13种抗菌药物的耐药率明显高于非产ESBLs株。鲍曼不动杆菌的产ESBLs株有较高的交叉耐药性,采用纸片确证实验检出ESBLs对于临床用药有一定的指导意义。     

Reliability of air displacement plethysmography

The purpose of this study was to examine the reliability of an air displacement plethysmography device (BOD POD) over trials performed on 3 different days. Subjects consisted of 24 healthy adults (8 men, 16 women), ages 18-38 years, with body weights 46.8-93.6 kg, body mass indexes of 19.1-30.1 kg x m(-2), and percentage body fats (BF) of 7.9-43.1%. Two estimates of BF were performed on 3 days. Paired t-tests revealed no significant within-day differences in body volume (BV), thoracic gas volume (V(TG)), body density (BD), and BF. Correlations between the two V(TG) measures on a day were r = 0.86 for day 1, r = 0.93 for day 2, and r = 0.96 for day 3. BF estimates within a day had high correlations of r = 0.98. Significant differences were found between days for measures of BV, V(TG), BD, and BF. These results indicate a high reliability for within-day estimates of BF and significant differences in between-day estimates of BF using air displacement plethysmography. Reliability of BF may be increased by requiring subjects to practice the procedure for V(TG) measurement.     

A mitogenic factor, released by stimulated human mononuclear cells and distinct from interleukin 2 (IL 2), B cell growth factor (BCGF), and interleukin 1 (IL 1)

Two lymphocyte mitogenic factors, interleukin 2 (IL 2) and blastogenic factor (BF), are generated concomitantly in human mixed lymphocyte cultures (MLC). The latter mitogenic factor is directly mitogenic for unstimulated lymphocytes, whereas the former mitogenic factor acts only on previously activated lymphocytes. Both factors had a m.w. range, as determined by gel filtration, of 18,000 to 30,000. Thus, these two factors were inseparable on the basis of m.w. size. However, BF and IL 2 were separable during ion exchange chromatography on the DEAE cellulose and phenyl-Sepharose chromatography. In addition, BF activity in the supernatants of MLC reached a maximum after day 5, whereas IL 2 activity peaked at day 3, thus distinguishing BF from IL 2 kinetically. These results clearly indicate that BF activity is mediated by molecules distinct from IL 2. The biochemical relationship between B cell growth factor (BCGF) and BF was also examined. Because BF was readily separable from BCGF by Con A-Sepharose chromatography, BF is distinguishable from BCGF. No augmentation of PHA-stimulated C3H mouse thymocyte proliferation was associated with the preparation of partially purified BF, demonstrating that BF and IL 1 are distinct molecules. Taken together, these results indicate that BF is clearly distinct from IL 2, BCGF, and IL 1. BF-containing MLC supernatants have direct mitogenic activity on both T and B cells. Both T and B cell blastogenic activities copurified during ammonium sulfate precipitation, gel filtration, DEAE cellulose ion exchange chromatography, and hydrophobic chromatography. Thus, these two activities appear to be biochemically inseparable. Monoclonal anti-Tac, that has been suggested to recognize the receptor for human IL 2, was highly inhibitory to the T cell response to the phenyl-Sepharose preparations of BF (IL 2-free). In contrast, this antibody had minimal or no effect on BF-induced B cell proliferation. However, when MLC supernatants were absorbed with a cloned IL 2-dependent T cell line, only IL 2 activity, but not BF activity, was removed, demonstrating that BF and IL 2 have different binding specificities. The precise mechanism(s) by which anti-Tac inhibits BF-induced proliferation of T cells is unknown at present. Additionally, during the course of these experiments, we observed that Con A-Sepharose chromatography could be used as a simple one-step method of separating BCGF from IL 2.     

机械通气患儿气管导管表面细菌生物膜形成及病原分析

目的观察儿科重症监护病房（PICU）机械通气（mechanical ventilation,MV）患儿气管导管（endotraeheal tube,mr）表面细菌生物膜（biofilm,BF）内细菌分布情况及BF形态学特征。方法以我院治疗的35例MV患儿为研究对象。收集第1次拔除或更换的ETT,经碘化丙啶（PI）和异硫氰酸荧光索标记刀豆蛋白A（FITC-ConA）染色后,激光共聚焦显微镜观察ETT-BF内细菌及胞外多糖（EPS）分布情况,并收集ETT表面和下呼吸道分泌物进行细菌分离、培养和鉴定。结果（1）35例MV患儿中,E1-r表面细菌培养阳性31例（88．57％）;17例EIT表面和下呼吸道分泌物同时分离出相同菌种,占ETT培养阳性的54．83％。在EIT-BF和下呼吸道分泌物中以金黄色葡萄球菌,肺炎克雷伯菌,大肠埃希菌,阴沟肠杆菌最常见。（2）CLSM观察可见,气管插管12h后,ETT-11表面已出现细菌黏附;48h时ETT表面可见大量短棒状或球状细菌黏附聚集,EPS较12h时显著增多,初步形成BF结构;72h左右细菌粘连成团块状,被EPS包裹,可见成熟BF结构形成;7d后细菌粘连成大片状,在其周围可见散在的微菌落。（3）35例MV患儿中,19例发生了呼吸机相关性肺炎（ventilated—associated pneumonia,VAP）。其中经口插管10例,经鼻插管9例。结论MV时细菌极易在ETT表面黏附,形成细菌BF。ETF表面细菌定植及BF形成与长时间MV患儿伴发呼吸机相关性肺炎之间可能存在一定相关性。     

Evaluation of an automated system for the detection of carbapenem resistant Acinetobacter baumannii and assessment of metallo-β-lactamase production using two different phenotyping methods

We tested 200 clinical Acinetobacter baumannii isolates against imipenem and meropenem using the methods of broth microdilution, disk diffusion, agar dilution, MicroScan/WalkAway automated system. Very major errors were mostly obtained between MicroScan/WA system and disk diffusion test. MicroScan/WA system generated unacceptable errors. Combined disk and modified Hodge tests used for detection of metallo-β-lactamase production were practical and useful.     

广州地区不动杆菌耐药调查

目的：了解广州地区不动杆菌属的耐药情况,指导临床合理用药。方法：对广州地区12家医院1998年至2001年从临床分离的436株不动杆菌属,严格按照NCCLS规定的标准用纸片扩散法进行药敏试验。结果：对12种抗生素的耐药监测发现,不动杆菌属对亚胺培南、头孢哌酮／舒巴坦、氨苄西林／舒巴坦、替卡西林／克拉维酸和哌拉西林／三唑巴坦耐药率分别为2．8％、5．6％、14．6％、14．6％和27．0％,对其他抗生素耐药率较高。结论：本地区不动杆菌耐药现象较普遍,应引起临床高度重视。