烧伤创面铜绿假单胞菌的分离和耐药谱分析

目的　探讨烧伤创面铜绿假单胞菌耐药谱的变迁及诱导型 β 内酰胺酶的产生。 方法对笔者医院 1996年 6月～ 2 0 0 0年 6月鉴定的 4 5 2株烧伤创面铜绿假单胞菌 ,用VITEK AMS全自动细菌鉴定及药敏系统、E test浓度梯度法做药敏试验 ,以K B法为基础作诱导酶的检测。　结果　 4年中临床常用的头孢菌素及亚胺培南的耐药率均有提高 ;其中以头孢哌酮 /舒巴坦的耐药率最低 ,但 4年中耐药率的变化差异也有显著性意义 (P <0 .0 5 ) ,亚胺培南的耐药率在 2 0 %～ 4 0 %之间。 12 0株野生型铜绿假单胞菌中 ,以亚胺培南为诱导剂 ,检出产生诱导酶者为 72 .5 %。　结论　烧伤创面铜绿假单胞菌耐药谱的分析和诱导酶的检测 ,可以及时监控和调整抗生素的使用 ,对避免铜绿假单胞菌在医院内流行具有重要意义     

院内不同时间段铜绿假单胞菌的耐药性分析

目的：分析本院内不同时间段分离的铜绿假单胞菌对各种抗菌药物的耐药性变化,为临床合理使用抗菌药物提供依据。方法对2012年10月至2012年12月（2012年第4季度）分离的142株铜绿假单胞菌、2013年1月至2013年3月（2013年第1季度）分离的129株铜绿假单胞菌、2013年4月至2013年6月（2013年第2季度）分离的74株铜绿假单胞菌、2013年7月至2013年9月（2013年第3季度）分离的140株铜绿假单胞菌,用WalkAway 96 PLUS NC50药敏板检测菌株对亚胺培南等12种抗菌药物的耐药性,并对检测结果分别进行分析。结果2012年第4季度分离的142株铜绿假单胞菌对替卡西林/克拉维酸、氨曲南和亚胺培南的耐药率分别为35.2%（50/140）、31.0%（44/142）和30.3%（43/142）,对哌拉西林、左旋氧氟沙星、环丙沙星、哌拉西林/他唑巴坦、头孢他啶、妥布霉素、庆大霉素、头孢吡肟和阿米卡星的耐药率为9.2%～26.8%。2013年第1季度分离的129株铜绿假单胞菌对替卡西林/克拉维酸、氨曲南、亚胺培南、左旋氧氟沙星为33.3%～44.4%,对环丙沙星、哌拉西林、头孢他啶、头孢吡肟、哌拉西林/他唑巴坦、庆大霉素、妥布霉素和阿米卡星的耐药率为9.3%～27.9%。2013年第2季度分离的74株铜绿假单胞菌对亚胺培南、替卡西林/克拉维酸、氨曲南、环丙沙星、左旋氧氟沙星和哌拉西林的耐药率为32.8%～47.3%,对妥布霉素、庆大霉素、头孢他啶、头孢吡肟、哌拉西林/他唑巴坦和阿米卡星的耐药率为17.6%～27.0%。2013年第三季度分离的140株铜绿假单胞菌以上12种抗菌药物的耐药率为2.9%～26.4%。不同阶段分离的菌株80%以上都来自痰液。结论院内不同季度分离的铜绿假单胞菌对临床常用抗菌药物的耐药率都存在一定的差异,2013年第1季度和第2季度分离的铜绿假单胞菌的耐药性显著高于2013年第3季度和2012年第4季度分离的铜?     

ICU铜绿假单胞菌耐药性调查

铜绿假单胞菌是医院感染的主要病原菌 ,该菌具有天然耐药性 ,增长迅速。为动态观察该菌的耐药性 ,2 0 0 0年 1月至 2 0 0 2年 2月采集ＩＣＵ病人呼吸道分离的 2 33株标本进行了耐药性调查。1　方法1 1　标本留取方法指导病人清晨先用朵贝尔液漱口 ,再用清水漱口后 ,用力咳出气管深处痰液入无菌培养盒中 (避免混入唾液及鼻腔分泌物 )送检。1 2　试验方法细菌鉴定用常规法 ,药敏试验采用纸片扩散法[1] 。纸片选择、试验方法、质量控制与判定标准按美国 1999年颁布的临床实验室标准委员会 (ＮＣ ＣＬＳ)标准。所用 14种药敏纸片是 :复方新诺明…     

假单胞菌JW12发酵液的脂肪酶稳定性

通过对假单胞菌（Ｐｓｅｕｄｏｍｏｎａｓｓｐ．）ＪＷ１２发酵液脂肪酶稳定性的研究，表明了发酵液脂肪的稳定性主要受蛋白酶的影响。当用吐温８０作为碳源时，通过对发酵培养基组成的调整，达到降低蛋白酶的目的，提高发酵液脂肪酶酶活。     

假单胞菌JW12脂肪酶的补料分批发酵

研究了假单胞菌ＪＷ１２脂肪酶在２Ｌ和２５Ｌ容积发酵罐的补料分批发酵工艺．通过调整碳源补加速率，控制产酶期发酵液ＰＨ在８．２左右，能有效提高脂肪酶的酶活和表观生产率．在２５Ｌ标准发酵罐中，连续补加吐温－８０，最高脂肪酶酶活为１２９．２μｍｏｌ／（ｍｉｎ·ｍＬ），表观生产率为１５．９１     

沼泽红假单胞菌的培养和生产番茄红素的优化控制

围绕沼泽红假单胞菌的高效培养和生产番茄红素的优化控制条件进行研究,结果表明,乙酸钠、双乙酸钠、柠檬酸三钠或丙酸钠均可以作为唯一有机碳源支持沼泽红假单胞菌的生长,培养5 d后细胞浓度均可达到6×109/mL以上;如果采用乙酸钠作为碳源,添加0.1 g/L山梨酸钾和10 g/L氯化钠可以促进沼泽红假单胞菌的生长,培养5 d细胞浓度高达8×109/mL以上.进一步研究发现以丙酸钠为惟一有机碳源时,培养出的沼泽红假单胞菌细胞番茄红素质量分数最高达6.66 g/kg,比生长于乙酸钠的提高了77.8%.     

铜绿假单胞菌感染在临床上耐药性的分析

目的:了解铜绿假单胞菌感染在临床上的耐药性及临床分布特点,为临床合理使用抗生素提供理论依据.方法:从本院各科室收集的标本中分离出98株铜绿假单胞菌,同时进行抗生素敏感试验.结果:铜绿假单胞菌在15种临床常用抗生素中,表现为多重耐药,其中3种抗生素的耐药率达到80％以上,而临床上以呼吸道感染为主,在各科室分布中以重症监护病房最高.结论:加强对呼吸道患者的防护,合理使用抗生素,对感染患者采取积极的防治措施,最大限度地降低铜绿假单胞菌的感染.     

2009至2011年铜绿假单胞菌的临床分布及耐药分析

目的：分析2009至2011年本院临床分离出来的铜绿假单胞菌耐药性的变迁,为临床合理的给药方案提供参考。方法采用肉汤稀释法对临床分离出的2781株病原菌中的252株铜绿假单胞菌对12种抗菌药物进行药敏分析。结果临床上分离铜绿假单胞菌标本主要为痰和支气管灌洗液下呼吸道标本,占62.3%（157/252）。铜绿假单胞菌感染发生率主要分布在ICU 27.8%（70/252）和感染科19.8%（50/252）。铜绿假单胞菌对大多抗菌药物的耐药性逐年增加,尤其亚胺培南和美洛培南已经达到41%（30/74）和35%（22/74）,敏感性最高的为多粘菌素B和氨基糖苷类阿米卡星。结论铜绿假单胞菌感染有上升趋势,对常用抗生素耐药率增加,需引起临床重视。加强其耐药监测可以为临床提供治疗依据。     

假单胞菌脂肪酶非水相催化性质

以丁酸与丁醇的酯化为模型反应，研究了假单胞菌脂肪酶的非水相催化性质。有机相中酶的催化作用需要一定的水分，最适水含量因溶剂的不同而有所差异。通常，极性强的溶剂其最适水含量高；当以水活度衡量时，各溶剂间的差别消失，最适水活度均在０．５～０．６之间。有机溶剂中酶的热稳定性较水溶液中明显提高，在环己烷中８０℃保温６ｈ酶活力仍保留约８０％；连续使用５次，酶活损失不到１０％．在环己烷中酶的最适作用温度为５０℃，比水溶液中略高；最适作用ｐＨ为９．０，与水溶液中相近。     

重症监护病房铜绿假单胞菌感染的危险因素和耐药性分析

目的 研究重症监护病房(icu)铜绿假单胞菌感染的发生率、危险因素及耐药情况.方法 收集171例脓毒症患者,分离培养铜绿假单胞菌并进行药敏试验.采用spss10.0统计学软件对可能的危险因素进行logistic回归分析.结果 171例脓毒症患者中,发生铜绿假单胞菌感染37例,分离培养出 45株铜绿假单胞菌.多因素logistic回归分析显示,近期抗菌药物使用史(or=4.291,95%ci:1.727～10.662)、住icu时间(or=1.117,95%ci:1.058～1.181)、机械通气(or=3.400,95%ci:1.348～8.579)以及中心静脉导管的使用(or=3.339,95%ci:1.322～8.434)为铜绿假单胞菌感染的独立危险因素.药敏试验显示铜绿假单胞菌对头孢寨肟的耐药率最高(68.9%),18株(40%)菌株表现为多重耐药.结论 铜绿假单胞菌足icu常见的病原菌,具有多重耐药性.加强抗菌药物的合理使用、严格执行各种有创导管的无菌操作是减少铜绿假单胞菌耐约的重要措施. abstract： objective to investigate the incidence, risk factors and drug-resistance of pseudomonas aeruginosa infection in intensive care unit (icu). methods totally 171 patients with sepsis admitted in icu were enrolled. pathogenic bacteria culture and antimicrobial susceptibility tests were performed. spss10. 0 software was used for logistic regression analysis of the risk factors. results pseudomonas aeruginosa infection was confirmed in 37 patients, and 45 strains of pseudomonas aeruginosa were isolated. logistic regression revealed that recent antibiotics use ( or = 4. 291 , 95% ci: 1. 727-10. 662) , length of icu stay (or = 1.117, 95% ci: 1.058-1. 181) , mechanical ventilation (or = 3.400, 95% ci: 1.348-8.579) and central venous catheterization (or =3. 339, 95% ci: 1.322-8.434) were independent risk factors of pseudomonas aeruginosa infection. the resistance rate of cefotaxime was the highest (68.9%) and 18 strains (40%) were multidrug-resistant. conclusions pseudomonas aeruginosa infection is common in icu and it is usually multidrug resistant. the rational use of antibiotics and aseptic technique of invasive catheterization are important for the prevention of pseudomonas aeruginosa infection.     

鼠李糖脂分批式发酵动力学模型

根据3.7 L瑞士Bioengineer KLF2000型发酵罐分批发酵的实验数据,利用GraphPad Prism软件对假单胞菌BS-03产鼠李糖脂生物表面活性剂的发酵动力学模型进行非线性拟合,说明Logistic模型和L-P模型能较好的描述假单胞菌BS-03发酵过程中的菌体生长、鼠李糖脂合成和限制性基质的消耗动力学。     

萘降解细菌的分离及其降解和转座基因的分子检测

用富集培养法，从工业废水和活性污泥中分离到9个高效降解萘的细菌菌株（ND7～ND15）．对菌株ND7、ND8、ND9和ND10所进行的16SrDNA序列分析表明，它们都属于假单胞菌属（Pseudomonas）．PCR实验结果表明，上述4个菌株都含有蔡降解基因nahAc、nahG、nahH和catA，ND7和ND8菌株还含有萘降解基因nahU.DNA杂交实验结果表明，上述9个萘降解菌株都含有转座酶基因tnpA1和解体酶基因tnpR．这些结果表明，萘降解细菌的降解基因和转座基因具有高度的保守性．酶学实验证明，ND7、ND8、ND9和ND10菌株都具有儿茶酚1，2-双加氧酶活力和儿萘酚2，3-双加氧酶活力，但在不同菌株中这两种酶的比活力有明显不同．     

乳酸杆菌肽聚糖的分离鉴定及其免疫活性

研究了乳酸杆菌细胞壁肽聚糖的分离、鉴定及其对肽聚糖生物活性的影响.实验获得了SDS处理粗细胞壁的适宜条件为SDS质量浓度8g/dL,处理时间为沸水浴10min.SDS结合胰蛋白酶和TCA处理,可有效去除乳酸杆菌中的非共价结合蛋白质及共价结合蛋白质.经化学分析鉴定,所提取的乳酸杆菌成分为肽聚糖,肽聚糖中丙氨酸、天冬氨酸、谷氨酸和赖氨酸浓度分别为1.181,0.943,0.770和0.456mmol/g,是肽聚糖中的组分氨基酸.SDS结合TCA处理,方能较有效地去除DNA.TCA处理是去除细胞壁磷壁酸、脂磷壁酸的有效方法.此外,小白鼠腹腔巨噬细胞的吞噬实验、C3b受体实验以及血清溶菌酶活力实验证实,所分离纯化肽聚糖的免疫学活性未受影响.     

Host range and geographical distribution of Babesia sp. Mymensingh

Bovine babesiosis represents a serious threat to the cattle industry in the tropics and subtropics. Although several Babesia species infect cattle, only B. bovis, B. bigemina and B. divergens are known to cause clinical babesiosis. However, our recent study demonstrated that the newly discovered Babesia sp. Mymensingh might be a virulent species capable of causing clinical babesiosis in cattle. The objective of this study was to determine the host range and geographical distribution of Babesia sp. Mymensingh on a global scale. A total of 2,860 archived DNA samples from 2,263 cattle in Sri Lanka (n = 672), the Philippines (n = 408), Vietnam (n = 460), Uganda (n = 409), Brazil (n = 164) and Argentina (n = 150); 419 buffalo in Sri Lanka (n = 327) and Vietnam (n = 92); and 127 goats and 51 sheep in Vietnam were screened using a Babesia sp. Mymensingh‐specific PCR assay. Babesia sp. Mymensingh infection was detected in cattle, buffalo, sheep and goats. Cattle of all countries surveyed in this study except Brazil were found to be infected with Babesia sp. Mymensingh. The highest positive rates were recorded in cattle from the Philippines (11.3%) and Vietnam (9.6%), followed by Argentina (4.7%), Sri Lanka (1.5%) and Uganda (1.0%). Buffalo were found to be infected with this parasite in Sri Lanka (1.2%) and Vietnam (10.9%). Unexpectedly, Babesia sp. Mymensingh was also detected in sheep (2.0%) and goats (1.3%) from Vietnam. These findings were confirmed by PCR amplicon sequencing. In conclusion, our present findings indicate that Babesia sp. Mymensingh, which infects cattle, buffalo, sheep and goats, is endemic in Asia, Africa and South America.     

Effect of Staphylococcus epidermidis on adherence of Pseudomonas aeruginosa and Proteus mirabilis to polymethyl methacrylate (PMMA) and gentamicin-containing PMMA

The goals of this study were to determine 1) effect of Staphylococcus epidermidis adherence and biofilm production on adherence of the opportunistic pathogens Proteus mirabilis and Pseudomonas aeruginosa to polymethyl methacrylate (PMMA); 2) if the biofilm killed by autoclaving altered adherence of other organisms; 3) if adherence of S. epidermidis to gentamicin-containing PMMA altered adherence of the opportunistic pathogens P. mirabilis and P. aeruginosa to gentamicin-containing PMMA. Results show that biofilms formed by S. epidermidis, whether alive or dead, significantly increased adherence of Pseudomonas. Adherence of Proteus was significantly increased on dead biofilms and increased, but not significantly (p = less than 0.1), on live ones. Greatest adherence seen in the study was to autoclaved biofilms. Significant adherence of Proteus and Pseudomonas was found on gentamicin-containing PMMA specimens, which were preincubated with S. epidermidis for formation on the biofilm. These results indicate that a biofilm is formed on PMMA-gentamicin specimens and this may impair the ability of gentamicin to kill other organisms.     

Severity and predicted outcome of postoperative Pseudomonas aeruginosa infections

The severity and predicted outcome of postoperative Pseudomonas aeruginosa (P. aeruginosa) infections (PPAI) was evaluated using a severity scoring system based on a simplification and modification of the APACHE II system. A total of 86 patients in whom P. aeruginosa was isolated from various sources were examined. PPAI developed in 50 patients, resulting in an overall mortality rate of 24%. An increased severity score (SS) correlated with an increased risk of developing PPAI. Thus, PPAI developed in 33% of the patients with an SS of 0–1, in 66.7% of those with an SS of 2–3, and in 100% of those with an SS of 6 or higher. Moreover, the mortality rate of the patients with an initial score of 6 or higher was 50%. The mean (±SD) initial severity score was 5.4±2.9 for survivors and 2.9±2.6 for nonsurvivors (P<0.01). In the patients who subsequently died, the SS remained high throughout the clinical course despite therapy, whereas in the survivors the SS decreased progressively, reflecting a favorable clinical course. These results suggest that our severity scoring system was useful for predicting outcome and monitoring the response of PPAI to therapy.     

枯草杆菌Bacillus sp F26产过氧化氢酶的发酵条件

从内蒙古呼伦贝尔草原的盐碱湖中分离到的一株低度嗜盐嗜碱细菌Bacillus sp F26，能积累高水平过氧化氢酶（CAT）。对Bacillus sp F26发酵产过氧化氢酶的环境与营养条件的研究结果表明，其积累高水平过氧化氢酶的适宜环境条件为：温度37℃，种龄20-22h，接种量5％，装液量50mL／（250mL的摇瓶）。适宜发酵培养基组成（g／L）为：葡萄糖15，牛肉膏10，玉米浆10，酵母膏5，磷酸二氢钾1，氯化镁0．2，氯化钠50，碳酸钠10。采用上述条件进行摇瓶分批发酵实验，发酵20h，过氧化氢酶酶活达到16．32U／mL，细胞干重为4．12g／L。进一步研究发现，在对数生长后期（16h）添加2mmol／L的H2O2可以明显刺激产酶，在5L的发酵罐上进一步以指数速率方式流加H2O2，由于该流加方式可降低H2O2对细胞的毒害作用，过氧化氢酶酶活达到29．89U／mL，与分批发酵相比提高了92．8％。     

小鼠透明带结合蛋白sp38在大肠杆菌中的融合表达和纯化

目的　表达及纯化小鼠透明带结合蛋白sp3 8。方法　从BALB/C小鼠睾丸组织提取总RNA ,利用RT PCR技术扩增sp3 8基因编码区序列 ,并将其重组到含麦芽糖结合蛋白 (MBP)的融合蛋白原核表达载体 pMAL c2x中 ,在大肠杆菌中进行表达。结果　序列测定表明克隆获得的sp3 8全长与基因库所登录的序列一致。经IPTG诱导表达出的MBP sp3 8融合蛋白分子量约 88kD ,经Westernblot分析证实。经直链淀粉琼脂糖凝胶 (amyloseresin)亲和层析和SephadexG 10 0分子筛层析纯化后 ,MBP sp3 8纯度达 90 %。 结论　成功地获得sp3 8蛋白 ,为进一步研究其生物学性能和用于免疫避孕的可能性打下了基础     

Effects of pulsatile perfusion pressure and storage on hearts preserved for 24 hours under hypothermia, for transplantation.

Canine hearts preserved for 24 hours under hypothermic pulsatile perfusion at a systolic pressure of 25 mm Hg had better perfusion and transplantation survival results than hearts perfused at 50 or 80 mm Hg. Also, hearts perfused at a systolic pressure of 25 mm Hg did better than simple hypothermically stored hearts or fresh allografts. These findings indicate that hearts are adequately perfused for 24 hours under hypothermia for transplantation at a systolic pressure of 25 mm Hg.     

Management of a multidrug-resistant Pseudomonas aeruginosa infected total knee arthroplasty using colistin. A case report and review of the literature

Multidrug-resistant infections present a serious clinical and therapeutical problem. Colistin is an old-used polymyxin with rather poor pharmacokinetic profile and a remarkable nephrotoxicity. However, the emergence of multidrug-resistant bacteria has recently led to the increased use of colistin as a potentially available therapy. This article presents a 75-year-old diabetic woman with an early onset total knee arthroplasty infection by a multidrug-resistant Pseudomonas aeruginosa bacterial isolate that was managed successfully with surgical removal of the knee prosthesis, antibiotic impregnated cement and intravenous administration of colistin for 6 weeks, and second stage revision knee surgery. Two years later, laboratory and imaging studies showed no evidence of recurrence of infection.