北京地区实验动物绿脓杆菌分离株MLVA分型

目的通过多位点可变数目串联重复序列分析（multiple-locus variable-number tandem-repeat analysis,MLVA）分型方法,研究北京地区实验动物绿脓杆菌分离株基因型和分布情况。方法选择13个可变数目重复序列（variable-number tandem-repeat,VNTR）位点,对实验动物及设施中检测出的141株绿脓杆菌的基因组DNA进行重复序列扩增,所得指纹图谱使用BioNumerics软件进行聚类分析,绘制系统发育树和最小生成树（minimum spanning tree,MST）。结果所采用的13个VNTR位点能够对全部分离株进行有效分型。141株绿脓杆菌主要被分为了3个基因群,56个基因型。各群所占比例分别为A群82.3%,B群占12.8%,C群占5.0%,辛普森多样性指数为0.763。同一区域内相邻实验动物单位的绿脓杆菌分离株同源关系较远。结论 MLVA方法对绿脓杆菌具有很好的分型能力,能够有效的追踪绿脓杆菌的来源。北京地区实验动物中绿脓杆菌分离株基因型多态性丰富,但无地域性同源关系。     

哈尔滨市112株绿脓杆菌的血清学分型和耐药性测定

对从临床分离的112株绿脓杆菌进行系统鉴定后,血清学分型表明:6、2和3型分别占32.14％、15.18％、15.18％,为主要流行型,共占总分离株的62.50％。耐药性测定结果为:对10种抗生素5耐以上者占69.6％。其中对多粘菌素、妥布霉素、丁胺卡那霉素三种抗生素最为敏感,敏感率分别为100％、70.6％、86.5％。     

"绿脓杆菌-MSHA菌毛株菌苗"对医院感染及肿瘤的治疗研究

目的：报导使用牟希亚,郭雁群发明的“绿脓杆菌-MSHA菌毛株菌苗”,全面调查与激活人体的免疫保护状态,从而消除了癌症患者的免疫低下状态,不仅对防治严重致死性条件病原体医院感染有良好作用。同时对增加癌症患者的CR率和降低其PD率也发生着重要影响。如经“菌苗”参与治疗后,肺癌的条件病原体感染率由单纯化疗的40.89%降至15.91%,恶性淋巴瘤的感染率由37.21%下降至17.78%;临床CR（完全缓解率）分别从6.67%升至25.00和从44.19%升至66.87%;临床PD（疾病恶化）率则从15.56%降至4.55%和自11.63%降至0。急性白血病的感染率从94.12%降至58.82%。免疫状态也明显向健康状态转变。     

VITEK－IMSGNS系列药敏卡对~（162）株绿脓杆菌的药敏分析

本文就临床标本中分离的１６２株绿脓杆菌,用ＶＩＴＥＫ－ＩＭＳ全自动微生物系统的ＧＮＳ－ＰＡ等四种药敏卡,共测定２３种抗生素药敏试验．该菌对亚胺硫霉素的敏感度（敏感和中度敏感）最高９７．７８％,其次是哌拉西林、替卡西林、丁胺卡那霉素、妥布霉素、环丙氟哌酸、头孢他啶和头孢哌酮,复方新诺明等八种抗生素耐药率均在９０％以上。分析该菌对１０种抗生素药敏结果发现,几种常用抗绿脓杆菌药物的最小抑菌浓度（ＭＩＣ）值普遍增高,并结合绿脓杆菌生物微膜形成和该菌对β－内酰胺类抗生素抗性机制作了讨论。     

O139霍乱弧菌检测方法的研究─诊断血清的制备

本文以Ｏ１３９死菌免疫健康家兔,经吸收去除非特异性凝集素制成的特异性诊断血清,专供诊断霍乱弧菌Ｏ１３９之用。采用玻片凝集试验对５株Ｏ１３９菌株及１２６株肠道菌进行验证,在敏感性和特异性方面均获得满意的结果。     

链球菌溶血素“O”纯化的研究

乙型链球菌３２１７２ 株接种于含有月示蛋白胨的牛肉水培养基中培养。得到含有溶血素“Ｏ”的培养液，其溶血效价２５６ ＩＵ／ｍ ｌ。比活为１１．６ ＩＵ／ｍ ｇ 蛋白。经过超滤除去９５％ 的非目的蛋白，所得超滤截留液的溶血效价为２０４８ ＩＵ／ｍ ｌ，比活为３３５．７ ＩＵ／ｍ ｇ 蛋白。在此基础上用离子交换柱层析纯化，所得活性峰溶血效价为８１９２ ＩＵ／ｍ ｌ，比活为４３１１．５ ＩＵ／ｍ ｇ 蛋白。通过以上两步的分级纯化，溶血素“Ｏ”的比活提高了３７１ 倍。此纯化蛋白在ＳＤＳ－ＰＡＧＥ电泳中呈现一条染色带。     

关于“临夏鸵鸟（Struthio linxiaensis）”分类位置的再讨论

对“临夏鸵鸟Struthio linxiaensis Hou et al．,2005”骨架和蛋壳化石的重新观察表明,其骨架和蛋壳化石的一些特征有悖于鸵鸟（Struthio）的特征,因此将“临夏鸵鸟”建立一新属：东方鸵鸟Orientornis gen．nov．以区别于Struthio,并修订了属的特征;修订后的鸵鸟科化石包括：Namornis,Diamantornis,Struthio,Orientornis gen．nov．4属。     

“三伏贴”对慢性阻塞性肺病系统炎症的影响

目的：观察“三伏贴”对稳定期慢性阻塞性肺疾病（COPD）患者系统炎症的影响,并评价其有效性。方法：收集稳定期慢性阻塞性肺疾病患者60 例,按随机数字表随机分为“三伏贴”治疗组与安慰剂治疗组,每组30 例;在常规西医治疗的基础上,分别给予“三伏贴”和安慰剂贴敷治疗,分别在入伏、初伏、中伏及末伏进行4 次穴位贴敷,完成1 疗程治疗者共56 例。观察治疗前、1 个疗程结束时、治疗后3 个月、治疗后6 个月血清炎症因子的变化。应用酶联免疫吸附试验（ELISA）检测血清白细胞介素-8（IL-8）,免疫比浊法测定血清CRP,比较治疗前后两组间的炎症因子变化趋势,同时比较治疗前后两组间CAT 评分（COPD assessment test,CAT）、呼吸困难评分（MRC）及生存质量评分。结果：“三伏贴”组、安慰剂组血清IL-8、CRP随时间变化趋势不同;其中试验组血清炎症因子随时间呈逐渐下降趋势,但治疗后6 个月时炎症因子略有升高,而安慰剂组炎症因子水平呈逐渐升高趋势,治疗前试验组与安慰剂组炎症因子水平差异无统计学意义,治疗3个月后,两组患者IL-8、CRP水平差异即有统计学意义（P〈0.05）。试验组患者治疗后CAT 评分、MRC 评分较治疗前下降,生存质量评分升高,差异有统计学意义（P〈0.05）,而安慰剂组患者在治疗后CAT 评分、MRC评分较治疗前升高,差异有统计学意义（P〈0.05）,生存质量评分下降,差异有统计学意义（P〈0.05）。同一时间点,试验组较安慰剂组CAT 评分、MRC评分下降,生存质量评分明显升高,差异有统计学意义（P〈0.05）。结论：冬病夏治穴位敷帖可以改善COPD 患者生存质量,缓解症状,其作用机理可能与降低系统炎症相关因子有关。     

“8455”小麦植株化学成分与麦蚜（长管蚜,二叉蚜）种群消长的关系

通过对两种肥力条件下的“８４５５”小麦植株化学成分与麦长管蚜和麦二叉蚜种群消长关系的研究与分析,结果表明可溶性糖、含钾量、含水量是影响麦二叉蚜种群消长的主要因子,胱氨酸是影响麦长管蚜种群消长的主要因子．     

Pyocine Typing of Clinical Strains of Pseudomonas aeruginosa

A total of 954 clinical isolates of Pseudomonas aeruginosa were typed by their ability to produce pyocines. The strains of Pseudomonas were isolated from urines, bloods, sputa, stools, and miscellaneous infectious exudates or tissue of patients of the Mayo Clinic and four associated hospitals. About 80% of the typable strains could be grouped into three major pyocine types: A (30.9%), B (34.8%), and D (14.1%). These large groups could be divided into subtypes by using additional indicator strains. There was no significant difference in the distribution of types by either institutional or specimen source, except that urine specimens yielded the highest percentage of one type. By this procedure, 93% of all isolates could be typed. Repeated typing of serially transferred strains indicated that the procedure has a high degree of reliability. Several strains exhibited extreme fluctuation in inhibition pattern. The procedure is a simple and reliable method to monitor the patterns of nosocomial infections due to P. aeruginosa.     

Serological Typing of 31 Achromogenic and 40 Melanogenic Pseudomonas aeruginosa Strains

Thirty-one achromogenic and 40 melanogenic Pseudomonas aeruginosa strains were studied with 10 monovalent typing sera (3). Twenty-one of the achromogenic (67.7%) and seven of the melanogenic (17.5%) strains were agglutinated by one of the 10 typing sera. Ten achromogenic and 33 melanogenic strains were not agglutinated by any of the 10 typing sera. As far as this set of antisera is concerned, the typability of achromogenic and melanogenic P. aeruginosa strains appears to be much lower than that of the chromogenic, nonmelanogenic strains of the species reported previously.     

Characterization of EO-1 Strains (Pseudomonas kingii) Isolated from Clinical Specimens and the Hospital Environment

Ten strains of the pseudomonad designated EO-1 and named Pseudomonas kingii, which were isolated from clinical specimens and the hospital environment, are described.     

Epidemiology of Pseudomonas aeruginosa in a Burns Hospital: Surveillance by a Combined Typing System

For 3 months, 259 cultures of Pseudomonas aeruginosa isolated from nonpatient environmental sources and 262 cultures from 16 infected patients in the Intensive Care Unit (ICU) of Shriners Burns Hospital were typed by a combined system with a high degree of reliability. Sinks were major sources of environmental contamination. Serotypes 1 and 2 were the predominant types found in patients, and they were most prevalent among typable strains from sinks. Strain designations were made on the basis of similarities in data from serological and phage typing. All nontypable strains were typed by pyocin production. Two infected patients carried different strains of P. aeruginosa that remained the same type for 45 days, even though their beds in ICU were approximately 6 feet apart. Cross-contamination from patient to patient and spread of infection by nursing personnel were eliminated as major modes of transmission because nasopharyngeal swabs, hair samples, and hands of nursing staff were consistently negative. Splashing of water from contaminated sinks to fomites was suggested as a possible mode of transfer for this infectious agent.     

Comparisons of the Genomes of New Giant Phages Isolated from Environmental Pseudomonas aeruginosa Strains of Different Regions

To study the genome diversity of bacteriophages from geographically distant natural populations, new giant KZ-like Pseudomonas aeruginosa phages isolated in two different regions were compared with earlier known phages of three species (KZ, Lin68, EL). A broad spectrum of lytic activity was demonstrated for all KZ-like phages. Phages of the KZ species proved to be common in natural populations of various regions, while EL- and Lin68-related phages were extremely rare. Most KZ-related phages had unique DNA restriction patterns, but the differences between these were only minor, and the genomes did not contain nonhomologous fragments. The spectrum of capsid polypeptides proved to be conserved in each species, and was proposed as a character necessary and sufficient for express classification of phages with an accuracy of species. Phages isolated in different geographical regions showed no substantial difference. Some phages only slightly differing in DNA restriction pattern from KZ may be used to study the origin of KZ genes coding for orthologs of proteins of unrelated species (other phages, pathogenic bacteria, eukaryotes).     

绿脓杆菌分子分型方法研究进展

绿脓杆菌是一种常见的人畜共患机会致病菌,广泛存在于自然界,是造成实验动物污染和医院内感染的重要病原菌之一。分子分型方法是病原菌流行病学分析的重要手段,对于确定感染来源和途径,检测交叉污染和流行菌株方面非常有效。本文主要对绿脓杆菌分子分型方法的研究进展进行综述。     

Biofilm Filtrates of Pseudomonas aeruginosa Strains Isolated from Cystic Fibrosis Patients Inhibit Preformed Aspergillus fumigatus Biofilms via Apoptosis

Pseudomonas aeruginosa (Pa) and Aspergillus fumigatus (Af) colonize cystic fibrosis (CF) patient airways. Pa culture filtrates inhibit Af biofilms, and Pa non-CF, mucoid (Muc-CF) and nonmucoid CF (NMuc-CF) isolates form an ascending inhibitory hierarchy. We hypothesized this activity is mediated through apoptosis induction. One Af and three Pa (non-CF, Muc-CF, NMuc-CF) reference isolates were studied. Af biofilm was formed in 96 well plates for 16 h ± Pa biofilm filtrates. After 24 h, apoptosis was characterized by viability dye DiBAc, reactive oxygen species (ROS) generation, mitochondrial membrane depolarization, DNA fragmentation and metacaspase activity. Muc-CF and NMuc-CF filtrates inhibited and damaged Af biofilm (p<0.0001). Intracellular ROS levels were elevated (p<0.001) in NMuc-CF-treated Af biofilms (3.7- fold) compared to treatment with filtrates from Muc-CF- (2.5- fold) or non-CF Pa (1.7- fold). Depolarization of mitochondrial potential was greater upon exposure to NMuc-CF (2.4-fold) compared to Muc-CF (1.8-fold) or non-CF (1.25-fold) (p<0.0001) filtrates. Exposure to filtrates resulted in more DNA fragmentation in Af biofilm, compared to control, mediated by metacaspase activation. In conclusion, filtrates from CF-Pa isolates were more inhibitory against Af biofilms than from non-CF. The apoptotic effect involves mitochondrial membrane damage associated with metacaspase activation.     

Typing of Pseudomonas aeruginosa strains in Turkish cystic fibrosis patients

The majority of cystic fibrosis (CF) patients suffer from chronic respiratory infection with the opportunistic bacterial pathogen Pseudomonas aeruginosa. The virulence of P. aeruginosa is associated with the presence of various extracellular factors, like alginate, elastase, alkaline protease which contribute tissue destruction and assist bacterial invasion. Virulence factor production of P. aeruginosa strains isolated from 46 CF patients followed in two cities in Turkey was detected. Strains were compared genotypically by arbitrarily primed PCR. Antimicrobial susceptibilities to 12 antibiotics were determined by broth microdilution method. Evaluation of virulence factor results revealed that 95.8% of the strains were alginate, 71.7% elastase and 52.1% alkaline protease producers. AP-PCR analysis revealed 35 genotypes indicated almost a complete discrepancy among the strains. The most effective drugs were penems and quinolones. Among aminoglycosides amikacin was the most effective one and a high level resistance to beta lactams was observed. Alginate is the most important virulence factor in the chronic colonisation of CF patients with P. aeruginosa. No evidence for cross infection between patients and for relationship between phenotypes and genotypes of the strains was found.