关于三株新血清型菌株补充绿脓杆菌国际抗原分型系统(IATS)的建议

作者在研制绿脓杆菌(PA)国际抗原分型系统(IATS)的基础上,收集了600余株绿脓杆菌,经生物学特性鉴定,交叉凝集试验和凝集素吸收试验,筛选出 CMCC-10117、PA-87、NO.446、PA-26、PS-1135株菌株(其中 NO.446、PA-26和 PS-113 3株为同型)系 IATS 不能包括的新血清型菌株。根据其抗原特异性从中选取了3株加入 IATS 中,使 IATS 由17个血清型发展为20个血清型。以新选出的MCC-10117作为 IATS-18型,PA-87作为 IATS-19型,NO.446作为 IATS-20型,经用468株临床分离的绿脓杆菌分型考核,分型率为99.4%。     

白土密相输送技术的试验研究及工业应用(Ⅲ)白土输送系统的现场改造及工业应用效果(续完)

在试验装置优化实验数据结果的基础上,为工业装置的白土输送系统提出改造方案,包括下料松动形式、密相输送白土操作参数、防堵自保系统、增力器、粉尘回收及尾气净化.生产实践证明效果良好.     

壳牌粉煤气化工艺磨煤与干燥系统(CMD)的分析建模及应用

分析壳牌煤气化工艺磨煤与干燥系统(Coal Milling and Drying System,简称CMD)的特点和煤质特性对系统的相关参数的影响;对CMD系统的主要控制参数的计算方法进行阐述,采用工艺模拟软件Pro/II建立CMD系统的热量和物料平衡计算模型,结合示范项目的运行情况对模型的计算准确性进行了比较分析。     

纤维素三(3,5-二甲基苯基氨基甲酸酯)液相色谱固定相的制备及手性拆分应用

采用涂渍的方法,合成了纤维素三（3,5-二甲基苯基氨基甲竣酯）HPLC固定相,并自制手性柱,在高效液相色谱正相色谱条件下,理论塔板数达到59005m^-1,对称因子为0．95。该柱成功地分离了醇类、羧酸类、胺类、氨基酸类手性化合物以及外消旋体药物,走现出良好的手性分离性能,是目前最有效的液相色谱手性分离柱之一。     

新显色剂N-烯丙基-N′-(氨基对苯磺酸钠)硫脲与银显色反应的研究及应用

在氨性介质中 ,Trition X-1 0 0存在下 ,N-烯丙基 -N′-(氨基对苯磺酸钠 )硫脲 ( ASATu)能与银发生反应 ,生成一个配合比为 1∶ 1的茶色水溶性配合物 ,其最大吸收波长在 4 1 0 nm。研究了此反应的适宜条件 ,并建立了一个测定银的新方法。该方法已用于阳极泥和地质样品中银的测定     

Utilization of response surface methodology to optimize the culture media for the production of rhamnolipids by Pseudomonas aeruginosa AT10

Pseudomonas aeruginosa AT10 produced a mixture of surface‐active rhamnolipids when cultivated on mineral medium with waste free fatty acids as carbon source. The development of the production process to an industrial scale included the design of the culture medium. A 2⁴ full factorial, central composite rotational design and response surface modelling method (RSM) was used to enhance rhamnolipid production by Pseudomonas aeruginosa AT10. The components that are critical for the process medium were the carbon source, the nitrogen source (NaNO₃), the phosphate content (K₂ HPO₄/KH₂PO₄ 2:1) and the iron content (FeSO₄·7H₂O). Two responses were measured, biomass and rhamnolipid production. The maximum biomass obtained was 12.06 g dm^?3 DCW, when the medium contained 50 g dm^?3 carbon source, 9 g dm^?3 NaNO₃, 7 g dm^?3 phosphate and 13.7 mg dm^?3 FeSO₄·7H₂O. The maximum concentration of rhamnolipid, 18.7 g dm^?3, was attained in medium that contained 50 g dm^?3 carbon source, 4.6 g dm^?3 NaNO₃, 1 g dm^?3 phosphate and 7.4 mg dm^?3 FeSO₄·7H₂O. © 2002 Society of Chemical Industry     

铜绿假单胞菌3型O-特异性多糖-破伤风类毒素结合疫苗的制备及其免疫特性分析

目的研制铜绿假单胞菌3型O-特异性多糖(O-specific polysaccharide,O-SP)-破伤风类毒素(Tetanus toxoid,TT)结合疫苗,并分析其免疫特性。方法采用热酚水法提取铜绿假单胞菌国际抗原分型系统(International Antigen Typing System,IATS)3型菌株的脂多糖(Lipopolysacchride,LPS),去除类脂A后纯化O-SP,用1-氰基-4-二甲胺基吡啶四氟硼酸盐(CDAP)活化O-SP,己二酸二肼(ADH)作为连接臂,在碳二亚胺(EDAC)作用下,与TT结合制备O-SP-TT结合疫苗,检测其理化性质,并分析结合疫苗的免疫原性和免疫保护性。结果 O-SP收获量相当于水解LPS量的30%,其蛋白质和核酸含量均低于1.0%,O-SP衍化度为3.83%,O-SP与TT的结合率约为40%,多糖蛋白质比(w/w)为1∶2.18。O-SP-TT结合疫苗可刺激小鼠产生高效价的IgG抗体,与O-SP组相比,差异有统计学意义(P0.05);结合疫苗免疫小鼠能保护5 LD50和10 LD50活菌的攻击,小鼠免疫O-SP-TT后兔抗血清对3 LD50和5 LD50活菌攻击具有良好的保护作用。结论制备的铜绿假单胞菌3型O-SP-TT结合疫苗具有良好的免疫原性和免疫保护性,该疫苗有望成为防治IATS 3型铜绿假单胞菌感染的有效疫苗。     

Strategies to Tackle Antimicrobial Resistance: The Example of Escherichia coli and Pseudomonas aeruginosa

Traditional antimicrobial treatments consist of drugs which target different essential functions in pathogens. Nevertheless, bacteria continue to evolve new mechanisms to evade this drug-mediated killing with surprising speed on the deployment of each new drug and antibiotic worldwide, a phenomenon called antimicrobial resistance (AMR). Nowadays, AMR represents a critical health threat, for which new medical interventions are urgently needed. By 2050, it is estimated that the leading cause of death will be through untreatable AMR pathogens. Although antibiotics remain a first-line treatment, non-antibiotic therapies such as prophylactic vaccines and therapeutic monoclonal antibodies (mAbs) are increasingly interesting alternatives to limit the spread of such antibiotic resistant microorganisms. For the discovery of new vaccines and mAbs, the search for effective antigens that are able to raise protective immune responses is a challenging undertaking. In this context, outer membrane vesicles (OMV) represent a promising approach, as they recapitulate the complete antigen repertoire that occurs on the surface of Gram-negative bacteria. In this review, we present Escherichia coli and Pseudomonas aeruginosa as specific examples of key AMR threats caused by Gram-negative bacteria and we discuss the current status of mAbs and vaccine approaches under development as well as how knowledge on OMV could benefit antigen discovery strategies.     

绿脓杆菌及其鞭毛蛋白联合rhIL-12体外对慢性乙型肝炎患者细胞免疫功能的影响

目的探讨绿脓杆菌及其鞭毛蛋白联合rhIL-12体外对慢性乙型肝炎患者细胞免疫功能的影响,为rhIL-12作为佐剂应用于临床提供理论依据。方法取健康人及慢性乙型肝炎患者外周血单个核细胞(PBMCs),分别与绿脓杆菌、鞭毛蛋白、rhIL-12、绿脓杆菌+rhIL-12、鞭毛蛋白+rhIL-12孵育,ELISA法检测细胞培养上清中IFNγ的含量。结果绿脓杆菌和鞭毛蛋白组只诱导产生低水平的IFNγ,rhIL-12组诱导产生IFNγ的水平较阴性对照组显著提高;而绿脓杆菌+rhIL-12组、鞭毛蛋白+rhIL-12组诱导产生IFNγ的水平显著高于绿脓杆菌、鞭毛蛋白和rhIL-12组,差异均有统计学意义,且rhIL-12的作用呈剂量依赖性。结论绿脓杆菌及其鞭毛蛋白联合rhIL-12体外能显著提高健康人和慢性乙型肝炎患者PBMCs产生IFNγ的水平,增强其细胞免疫应答。     

Pseudomonas aeruginosa DnaK Stimulates the Production of Pentraxin 3 via TLR4-Dependent NF-κB and ERK Signaling Pathways

Microbe-derived factors trigger innate immune responses through the production of inflammatory mediators, including pentraxin 3 (PTX3). PTX3 is a soluble pattern recognition molecule that stimulates the clearance of clinically important bacterial pathogens such as Pseudomonas aeruginosa. However, the P. aeruginosa factors responsible for the production of PTX3 have not been elucidated. In this study, we found that P. aeruginosa DnaK, a homolog of heat shock protein 70, induced PTX3 production. Induction was mediated by intracellular signals transmitted through the Toll-like receptor 4 (TLR4) signaling pathway. Following receptor engagement, the stimulatory signals were relayed initially through the nuclear factor kappa B (NF-κB) signaling pathway and subsequently by extracellular signal-regulated kinases (ERK), which are mitogen-activated protein kinases. However, ERK activation was negatively controlled by NF-κB, implying the existence of negative crosstalk between the NF-κB and the ERK pathways. These data suggest that P. aeruginosa DnaK acts as a pathogen-associated molecular pattern to trigger modulation of host defense responses via production of PTX3.     

Isolation and Identification of Phenanthrene Degrading Bacteria from the Soil around Oil Company of Andimeshk and Investigation of Their Growth Kinetics

Polycyclic aromatic hydrocarbons are particularly important due to large distribution in the environment, high toxicity and their carcinogen and mutagen properties. The purpose of this study was isolation and identification of phenanthrene degrading bacteria from the soil around Oil Company of Andimeshk (Iran) and investigation of their growth kinetics. Sampling from three stations was done at two seasons, spring and summer. Phenanthrene degrading bacteria were isolated from soil using enrichment method. Bacterial identification was performed by biochemical tests and 16S rRNA gene sequencing. Minimum inhibitory concentrations (MIC) were determined at various concentrations of phenanthrene. Bacterial biodegradation rate was determined using HPLC analysis. Finally, the growth kinetics of resistant bacteria was determined with culturing at concentrations of 0.5–0.8 g/l of phenanthrene. According to biochemical and molecular tests Pseudomonas aeruginosa strain SBL, Bacillus cereus strain Z4B-11, Staphylococcus epidermidis, and Micrococcus luteus were identified as phenanthrene degrading bacteria. The results showed that P. aeruginosa SBL and B. cereus Z4B-11 with the greatest amount of MIC are the most phenanthrene resistant bacteria, respectively. These two strains degraded 70% and 50% of phenanthrene after one week of incubation, respectively. The most growth in different concentrations of phenanthrene belonged to P. aeruginosa SBL and B. cereus Z4B-11 while the least growth belonged to S. epidermidis and M. luteus, respectively. It could be concluded that two new strains SBL and Z4B-11 which were isolated in the soil around Oil Company of Andimeshk have relatively high potential to be used for bioremediation of phenanthrene.     

Whole Genome Sequencing and Tn5-Insertion Mutagenesis of Pseudomonas taiwanensis CMS to Probe Its Antagonistic Activity Against Rice Bacterial Blight Disease

The Gram-negative bacterium Pseudomonas taiwanensis is a novel bacterium that uses shrimp shell waste as its sole sources of carbon and nitrogen. It is a versatile bacterium with potential for use in biological control, with activities including toxicity toward insects, fungi, and the rice pathogen Xanthomonas oryzae pv.oryzae (Xoo). In this study, the complete 5.08-Mb genome sequence of P. taiwanensis CMS was determined by a combination of NGS/Sanger sequencing and optical mapping. Comparison of optical maps of seven Pseudomonas species showed that P. taiwanensis is most closely related to P. putida KT 2400. We screened a total of 11,646 individual Tn5-transponson tagged strains to identify genes that are involved in the production and regulation of the iron-chelator pyoverdine in P. taiwanensis, which is a key anti-Xoo factor. Our results indicated that the two-component system (TCS) EnvZ/OmpR plays a positive regulatory role in the production of pyoverdine, whereas the sigma factor RpoS functions as a repressor. The knowledge of the molecular basis of the regulation of pyoverdine by P. taiwanensis provided herein will be useful for its development for use in biological control, including as an anti-Xoo agent.     

国际录井作业防H_2S应急预案的编制与应用

录井现场的应急制度主要包括应急程序的编制和应急演练。结合国际录井作业经验,以防H2S应急预案为例,讨论了预案编制的目的、方法、步骤,介绍了预案的演练和实际应用,最后提出了预案更新和完善的重要性。     

Production of a polyclonal antibody to the VP26 nucleocapsid protein of white spot syndrome virus (wssv) and its use as a biosensor

White spot syndrome virus (WSSV) is a major cause of high mortality in cultured shrimp all over the world. VP26 is one of the structural proteins of WSSV that is assumed to assist in recognizing its host and assists the viral nucleocapsid to move toward the nucleus of the host cell. The objective of this work was to produce a polyclonal antibody against VP26 and use it as a biosensor. The recombinant VP26 protein (rVP26) was produced in E. coli (BL21), purified and used for immunizing rabbits to obtain a polyclonal antibody. Western blot analysis confirmed that the antiserum had a specific immunoreactivity to the VP26 of WSSV. This VP26 antiserum was immobilized onto a gold electrode for use as the sensing surface to detect WSSV under a flow injection system. The impedance change in the presence of VP26 was monitored in real time. The sensitivity of the biosensor was in the linear range of 160–160000 copies of WSSV, indicating that it is good and sensitive for analysis of WSSV. The specificity of the biosensor was supported by the observation that no impedance change was detected even at high concentrations when using Yellow Head Virus (YHV). This biosensor may be applied to monitor the amount of WSSV in water during shrimp cultivation.