Antibacterial activity of Lawsonia inermis Linn(Henna) against Pseudomonas aeruginosa

Objective:To investigate the antibacterial activity of henna(Lawsonia inermis Linn) obtained from different regions of Oman against a wide array of micro-organisms.Methods:fresh henna samples were obtained from different regions of Oman as leaves and seeds,100 g fresh and dry leaves and SO g of fresh and dry seeds were separately soaked in 500 mL of ethanol for three days,respectively,with frequent agitation.The mixture was filtered,and the crude extract was collected.The crude extract was then heated,at 48 ℃ in a water bath to evaporate its liquid content.The dry crude henna extract was then tested for its antibacterial activity using well-diffusion antibiotic susceptibility technique.Henna extracts were investigated for their antibacterial activity at different concentrations against a wide array of different micro-organisms including a laboratory standard bacterial strain of Pseudomonas aeruginosa(NCTC 10662)(A aeruginosa) and eleven fresh clinical isolates of P.aeruginosa obtained from patients attending the Sultan Qaboos University Hospital(SQUH).2-Hydroxy-p-Nathoqinone-Tech(2-HPNT, MW=174.16,C_(10)H_40_3) was included as control(at 50%concentration) along with the henna samples tested.Results:Henna samples demonstrated antibacterial activity against all isolates but the highest susceptibility was against P.aeruginosa with henna samples obtained from Al-sharqyia region.Conclusions:Omani henna from Al-sharqyia region demonstrates high in vitro anti-P. aeruginosa activity compared with many henna samples from different regions of Oman.

Mutational and Phylogenetic Analysis of <i>nfxB</i>Gene in Multidrug-Resistant Clinical Isolates of <i>Pseudomonas aeruginosa</i>Hyperexpressing MexCD-OprJ Efflux Pump

The present study focused on MexCD-OprJ efflux pump and its regulatory gene nfxB in multidrug resistant (MDR) clinical isolates of Pseudomonas aeruginosa collected from Kerala, South India. Semi-quantitative reverse transcription-PCR technique was employed to detect hyperexpression of the efflux pump gene, mexD. Amplicons from nfxB gene of isolates hyperexpressing the efflux pump were sequenced for mutational and phylogenetic analysis. Among 29 isolates of MDR P. aeruginosa, increased mexD transcription was detected in 10.3% of the isolates when compared with P. aeruginosa reference strain, PAO (MTCC-3541). Various synonymous and non-synonymous mutations in nfxB regulatory gene sequences were detected. Notably, mutations detected in the strains designate Pa6 and Pa7 have been found to be novel and are hitherto unreported in GenBank data base. The genetic divergence and homogeneity of the nfxB regulatory gene sequences of mexCD-oprJ operon were clearly apparent in the phylogram generated employing similar sequences retrieved from the public database.

Biodegradation of Petroleum Hydrocarbon by Pseudomonas Aeruginosa

A bacterial strain of the genus Pseudomonas aeruginosa was inoculated into a hydrocarbon culture medium and incubated for a definite period of time. The ability of the bacterial strain to biodegrade a hydrocarbon, viz. n-hexadecane, was evaluated through determining the hexadecane concentration in the inoculated culture medium on a gas chromatograph (GC). The effect of pH value on the degrading ability of the bacterial isolate and the impact of temperature on microbial growth were also explored. Test results showed that Pseudomonas aeruginosa was markedly effective in biodegrading n-hexadecane. Furthermore, the ability of Pseudomonas aeruginosa to biodegrade n-hexadecane was different at various pH values. Pseudomonas aeruginosa provided excellent degrading ability at a pH value of 7.0. The microbial cells of Pseudomonas aeruginosa increased with an increasing incubation duration at temperatures ranging from 28 ℃ to 35 ℃, and an exponential phase of microbial growth was observed.

Characterization of Cr(Ⅵ) resistance and reduction by Pseudomonas aeruginosa

The experiments were conducted to evaluate the Cr(Ⅵ)resistance and reduction by Pseudomonas aeruginosa.After this bacterium tolerated 40 mg/L Cr(Ⅵ),the growth of cells was observed.The bacterial growth was obviously lower than the controls over 24 h and the binary cell fission was observed in cell morphology by scanning electron microscope.P.aeruginosa was found to be able to reduce Cr(Ⅵ)although Cr(Ⅵ)had toxic effects on the cells.The results demonstrate that Cr(Ⅵ)is reduced from 40 mg/L to about 18 mg/L in 72 h.The value of pH drops from 7.02 to around 5.65 after 72 h.A significant increase in the value of redox potential occurs during Cr(Ⅵ)reduction and Cr(Ⅵ)reduction can be observed over a range of redox potential from+3 mV to+91 mV.Both of SO4 2-and NO3 -have no effect on Cr(Ⅵ)reduction.The presence of Zn 2+has a notable inhibitory effect on Cr(Ⅵ) reduction while Cu 2+ substantially stimulates Cr(Ⅵ)reduction.In the presence of Zn 2+ ,Cr(Ⅵ)decreases from 40 mg/L to only 26-27 mg/L,whereas Cr(Ⅵ)drops to 1-2 mg/L after 48 h in the presence of Cu2 +.

Other Possible Causes of a Well-Publicized Outbreak of <i>Pseudomonas aeruginosa</i>Following Arthroscopy in Texas

Background: Seven patients at a hospital in Houston, TX, were diagnosed during a two-week period in 2009 with joint space infection of pansusceptible P. aeruginosa following arthroscopic procedures of the knee or shoulder. Tosh et al. (2011), who investigated and published the principal report discussing this bacterial outbreak, conclude that its most likely cause was the improper reprocessing of certain reusable, physically-complex, heat-stable arthroscopic instruments used during these arthroscopic procedures. These reusable instruments reportedly remained contaminated with remnant tissue, despite diligent efforts by the hospital to clean their internal structures. This retained bioburden presumably shielded the outbreak’s strain of embedded P. aeruginosa from contact with the pressurized steam, reportedly resulting in ineffective sterilization of these arthroscopic instruments and bacterial transmission. Objectives: First, to clarify which specific sterilization methods, in addition to steam sterilization, Methodist Hospital employed to process its reusable arthroscopic instrumentation at the time of its outbreak, in 2009;second, to evaluate Tosh et al.’s (2011) conclusion that ineffective steam sterilization due to inadequate cleaning was the most likely cause of this hospital’s outbreak;third, to consider whether any other hitherto unrecognized factors could have plausibly contributed to this outbreak;and, fourth, to assess whether any additional recommendations might be warranted to prevent disease transmission following arthroscopic procedures. Methods: The medical literature was reviewed;some of the principles of quality assurance, engineering and a root-cause analysis were employed;and Tosh et al.’s (2011) findings and conclusions were reviewed and compared with those of other published reports that evaluated the risk of disease transmission associated with the steam sterilization of physically-complex, heat-stable, soiled surgical instruments. Results and Conclusion: Reports documenting outbreaks of P. aeruginosa or another vegetative bacterium associated with the steam sterilization of inadequately cleaned surgical or arthroscopic instruments are scant. This finding—coupled with a number of published studies demonstrating the effective steam sterilization of complex instruments contaminated with vegetative bacteria mixed with organic debris, or, in one published series of tests, with resistant bacterial endospores coated with hydraulic fluid—raises for discussion whether Methodist Hospital’s outbreak might have been due to one or more factors other than, or in addition to, that which Tosh et al. (2011) conclude was its most likely cause. An example of such a factor not ruled out by Tosh et al. (2011) findings would be the re-contamination of the implicated arthroscopic instruments after sterilization. The specific methods that Methodist Hospital employed at the time of its outbreak to sterilize some of its arthroscopic instrumentation remain unclear. A number of additional recommendations are provided to prevent disease transmission following arthroscopic procedures.

Effect of Antibiotic Consumption on Resistance of <i>Pseudomonas aeruginosa</i>Isolated from Lebanese Patients with Emphasis on MBL Production

The relationship between antibiotic consumption and resistance has been widely evaluated. Pseudomonas aeruginosa is one of the most important opportunistic pathogens in the nosocomial setting, and its resistance to antibiotics is increaseing. Production of metallo-β-lactamases (MBLs) is currently the most fearful resistance mechanism due to the potential of dissemination. This study aimed to evaluate the correlation between antibiotic consumption (expressed in DDD/100 bed days) and resistance (expressed in % of isolates and patients) in different time periods for P. aeruginosa between 2006 and 2009 at Saint George Hospital University Medical Center (SGH-UMC), Beirut. Pearson correlation coefficients (r) were calculated and linear regression was performed. Detection of MBL-producing Imipenem resistant P. aeruginosa (IRPA) isolates between 2008 and 2009 was performed using three MBL screening methods: MBL Etest?, Imipenem/EDTA combined disk test and EDTA disk potentiation with four cephalosporins. The modified Hodge test was also performed. From 2006 till 2009, there was a trend of increasing resistance of P. aeruginosa to all antibiotics, and the highest % of resistance was for Ofloxacin. Concerning resistance expressed by isolates, high correlation coefficients resulted among Imipenem, Ciprofloxacin and Tazobactam consumption and resistance to these agents in the same year correlation;Ceftazidime and Ofloxacin consumption and resistance in the next year correlation;Gentamicin and Ofloxacin consumption and the change in resistance (ΔR). Concerning resistance expressed by patients, results were similar except for Ceftazidime and Ofloxacin correlation in the next year correlation. In MBL screening, three isolates gave accordance among 4 methods which showed a positive result. The correlation between antibiotic consumption and resistance is highly dependent on the kind of antibiotic, the organism and the time of correlation. Various MBL screening phenotypic methods on one isolate can increase accuracy and eliminate false positive and negative results.

Antibiotic sensitivity pattern of bacterial pathogens in the intensive care unit of Fatmawati Hospital,Indonesia

Objective:To evaluate the sensitivity pattern of bacterial pathogens in the intensive care unit(ICU) of a tertiary care of Falmawati Hospital Jakarta Indonesia.Methods:A cross sectional retrospective study of bacterial pathogen was carried out on a total of 722 patients that were admitted to the ICU of Fatmawati Hospital Jakarta Indonesia during January 2009 to March 2010. All bacteria were identified by standard microbiologic methods,and(heir antibiotic susceptibility testing was performed using disk diffusion method.Results:Specimens were collected from 385 patients who were given antimicrobial treatment,of which 249(64.68%) were cultured positive and 136(35.32%) were negative.The most predominant isolate was Pseudomonas aeruginosa(P.aeruginosa)(26.5%) followed by Klebsiella pneumoniae(K.pneumoniae)(15.3%) and Staphylococcus epidermidis(14.9%).P.aeruginosa isolates showed high rate of resistance to cephalexin(95.3%),cefotaxime(64.1%),and ceftriaxone(60.9%).Amikacin was the most effective(84.4%) antibiotic against P.aeruginosa followed by imipenem(81.2%),and meropenem(75.0%).K.pneumoniae showed resistance to cephalexin(86.5%),ceftriaxone(75.7%),ceftazidime(73.0%),cefpirome(73.0%) and cefotaxime(67.9%),respectively.Conclusions:Most bacteria isolated from ICU of Fatmawati Hospital Jakarta Indonesia were resistant to the third generation of cephalosporins,and quinolone antibiotics.Regular surveillance of antibiotic susceptibility pallerns is very important for setting orders to guide the clinician in choosing empirical or directed therapy of infected patients.

Characterization and Identification of Two Opportunistic Human Bacterial Pathogens in Rice

Burkholderia cepacia （Bc） and Pseudomonas aeruginosa （Pa） are both biocontrol agents in agriculture and opportunistic human pathogens in hospitals. Effective management and utilization practice is needed to understand their characteristics and distribution in rice. During the last decade, the two opportunistic human pathogens were detected in 631 samples of rice seed and 117 samples of rice plant in plain, highland and mountainous rice growing areas of China. Bc and Pa were primarily differentiated by common bacteriological characteristics and pathogenic tests and then identified into species by Biolog and FAME tests. However, the genotypes of Bc still could not be distinguished. It has been noted that the Bc and Pa mainly existed in rice root with the highest distribution frequency in plain areas （ 6.1% and 16.1%） and lowest in the mountainous areas （1.0% and 7.8%）.

An Impact of Different Silicone Breast Implants on the Bacterial Attachment and Growth

Bacterial biofilms have been implicated with breast implant complications including capsular contracture, double-capsule formation, and breast implant-associated anaplastic large cell lymphoma. However, the relationship between implant surface texture and microbial biofilm formation is insufficiently evaluated. In the present study, we examined the antimicrobial activities of different types of silicone breast implant. The growth of bacterial including Staphylococcus aureus, Staphylococcus epidermidis, and Pseudomonas aeruginosa was compared using implants with various surface textures, including Hans Smooth, Hans SmoothFine, Allergan Smooth, Eurosilicone Smooth, Eurosilicone Texture, Sebbin Smooth, Sebbin Micro, Sebbin Texture, and Motiva Smooth. Microbial investigation revealed the increased growth of S. aureus on breast implants after 48 h, except Eurosilicone Smooth, Eurosilicone Texture, Hans SmoothFine and Sebbin Smooth material. At 48 hours, there was no major difference between the S. aureus attachment on smooth and textured implants. The results of S. epidermis attachment on the implant after 48 h showed that their growth decreased on surfaces of Motiva Smooth, Sebbin Smooth, and Eurosilicone Smooth. These results indicated that S. epidermis was unable to survive on these breast implants. Eventually, P. aeruginosa count had showed decrease of bacterial count after 48 hours compared to 24 hours in most of the implants except for Eurosilicone Texture, Sebbin Smooth and Sebbin Micro, where the count of P. aeruginosa slightly increased. This indicated that P. aeruginosa was unable to exist on the smooth surfaces. Our results show that the in vitro assay revealed no significant difference between smooth and textured surfaces and showed variable interactions and needed further molecular analysis to assess their adherence nature.

2020—2022年某医院主要细菌耐药情况分析

目的通过分析福建中医药大学附属第二人民医院2020年1月—2022年12月常见细菌的耐药性,了解医院的细菌耐药情况,为临床提供合理使用抗菌药物的参考依据。方法通过回顾性分析方法对2020年1月—2022年12月福建中医药大学附属第二人民医院患者标本分离的病原菌株进行统计,从病原菌标本分布、病原菌种类及耐药等方面进行分析。结果2020—2022年分离的菌株共计12009株,呼吸道标本(包括痰、肺泡灌洗液)最多。前5位为大肠埃希菌2139株(17.81%)、肺炎克雷伯菌1342株(11.17%)、铜绿假单胞菌1094株(9.11%)、金黄色葡萄球菌为主793株(6.60%)、鲍曼不动杆菌541株(4.50%)。耐碳青霉烯类肺炎克雷伯菌(Carbapenemresistant K.pneumoniae,CR-KP)和耐碳青霉烯类铜绿假单胞菌(Carbapenemresistant Pseudomonas aeruginosa,CR-PA)2022年比2020年明显升高,差异有统计学意义(P<0.01)。其余3种耐甲氧西林金黄色葡萄球菌(Methicillin-resistant S.aureus,MRSA)、耐碳青霉烯类大肠埃希菌(Carbapenemresistant Escherichia coli,CR-EC)、耐碳青霉烯类鲍曼不动杆菌(Carbapenemresistant Acinetobacter baumannii,CRAB)3年间检出率差异无统计学意义(P>0.05)。鲍曼不动杆菌对大多数常用抗菌药物耐药率>50%。结论福建中医药大学附属第二人民医院主要以革兰阴性杆菌为主,鲍曼不动杆菌对大多数常用抗菌药物耐药率>50%,CR-KP(耐碳青霉烯类肺炎克雷伯菌)和CR-PA(耐碳青霉烯类铜绿假单胞菌)的耐药率呈增高趋势,建议临床密切关注病原菌耐药情况的变化,减少多重耐药菌的产生。

Effect of Qiguiyin Decoction(芪归银方) on Multidrug-Resistant Pseudomonas aeruginosa Infection in Rats

Objective： To investigate the effect of Qiguiyin Decoction （芪归银方, QGYD） on multidrug-resistant Pseudomonas aeruginosa infection in Sprague-Dawley （SD） rats. Methods： A pseudomonal infection model in SD rats was established by injecting multidrug-resistant P. aeruginosa intraperitoneally. Infected rats were randomized into four groups treated with Pure water, QGYD, ceftazidime, or combined QGYD and ceftazidime. Blood samples were obtained from the abdominal aorta. Serum was then collected and analyzed by peptide array for immune responsiveness to multidrug-resistant beta-lactamase proteins, including Verona integronencoded metailo-beta-lactamase 1 （VIM-l）, Sao Paulo metallo-beta-lactamase 1 （SPM-1）, and Temoniera （TEMs）. Blood levels of interleukin-113 （IL-113）, interleukin-4 （IL-4）, and interferon-γ （IFN-γ） were assessed by enzyme-linked immunosorbent assay. Results： QGYD enhanced antibody reactivity against VIM-1 [epitopes 7-11 and 36-40] and TEM-1 [epitopes 26-27, 52-55, and 66-70]. QGYD treatment restored the compromised antibody reactivity against VIM-1 [epitopes 53-54 and 56-58] and SPM-1 [epitopes 16-19 and 82-85] following pseudomonal infection. Serum levels of IL-113 and Thl/＇l-h2 in the rats were significantly elevated following pseudomonal infection （P〈0.05 or P〈0.01）. In contrast, QGYD and combination QGYD and ceftazidime treatment restored the elevated serum IL-1β and Thl/-rh2 levels to normal （P〉0.05）. Conclusions： QGYD improves the immune response to pseudomonal infection in rats by stimulating the production of protective antibodies against drug-resistant proteins VIM-1, SPM-1, and TEM-1. In addition, it protects the immune system and maintains immune responsiveness by restoring IL-1β and Thl/Th2 levels.

麻杏石甘汤对铜绿假单胞菌肺炎大鼠细菌清除效应及中性粒细胞浸润和炎症因子的影响

[目的]观察麻杏石甘汤对铜绿假单胞菌(PA)肺炎大鼠细菌清除效应及炎症因子和中性粒细胞(PMNs)浸润的影响。[方法]选取雄性SD大鼠120只,随机分为5组(空白组、模型组、西药组、中药组、中西医结合组),每组24只,采用气管滴注法,建立肺炎模型,空白组和模型组予生理盐水灌胃,西药组肌注头孢他啶,中药组予麻杏石甘汤浓缩煎剂灌胃,中西医结合组采用肌注头孢他啶联合麻杏石甘汤浓缩煎剂灌胃,分4个时点(造模后0、12、24、72 h)处死大鼠,每组每个时点处死6只,取肺组织及腹主动脉血,观察肺组织病理改变、检测肺组织匀浆细菌计数、髓过氧化物酶(MPO)表达、外周血PMNs、血清肿瘤坏死因子-α(TNF-α)、白细胞介素-8(IL-8)、白细胞介素-10(IL-10)水平。[结果]1)造模后各组组内前后时点比较,模型组和药物干预组(西药组、中药组和中西医结合组)肺部炎症均逐渐改善,肺组织细菌计数均逐渐下降,差异具有统计学意义(P<0.05);72 h时点,药物干预组与模型组比较,肺部炎症均明显改善,肺组织细菌计数均降低,差异具有统计学意义(P<0.05)。2)外周血PMNs计数,模型组和西药组呈持续上升趋势,中药组和中西医结合组呈先上升后下降趋势,不同时点组间比较,差异无统计学意义(P>0.05)。3)肺组织MPO表达量,模型组呈先上升后下降趋势,药物干预组均呈下降趋势;72 h时点,药物干预组MPO表达量低于模型组,差异有统计学意义(P<0.05)。4)炎症因子比较,造模后72 h内,模型组和药物干预组血清IL-10水平均呈上升趋势,72 h时点,中药组和中西医结合组IL-10水平均高于模型组,差异有统计学意义(P<0.05);造模后0、12、24 h,模型组和药物干预组血清TNF-α、IL-8水平均呈上升趋势,24 h后,中药组和中西医结合组TNF-α呈下降趋势,药物干预组IL-8均呈下降趋势,72 h时点,药物干预组TNF-α水平较模型组均降低,差异具有统计学意义(P<0.05)。[结论]麻杏石甘汤可促进机体清除PA,减少肺组织PMNs浸润,调节血清炎症因子TNF-α、IL-8、IL-10水平,减轻肺部炎症损伤。

铜绿假单胞菌致病毒力基因的表达与耐药性相关分析

目的:探讨铜绿假单胞菌(PA)Ⅲ型分泌系统(T3SS)4种致病毒力基因exoT、exoY、exoS、exoU的表达情况及其与耐药性的相关性。方法:收集安徽医科大学附属安庆第一人民医院2022年9月~2023年9月临床检出的不同标本来源的PA 144株,使用Vitek2-Compact仪器进行细菌鉴定并分析药敏结果,PCR检测T3SS系统致病毒力基因exoT、exoY、exoS、exoU携带表达情况,分析不同致病毒力基因型组合与细菌耐药性之间的关系。结果:144株PA携带exoT(144/144)和exoY(144/144)基因的阳性率为100%,exoS基因占84.72%(122/144),exoU基因占20.14%(29/144)。总共检出4种致病毒力基因型组合,以exoT+/exoY+/exoS+/exoU-(79.17%,114/144)基因型组合为主,其次是exoT+/exoY+/exoS-/exoU+(14.58%,21/144)。药敏结果显示exoT+/exoY+/exoS-/exoU+基因型菌株对常见抗菌药物如妥布霉素、头孢他啶、左旋氧氟沙星、环丙沙星、哌拉西林、替卡西林/克拉维酸、美洛培南、粘菌素耐药率高于exoT+/exoY+/exoS+/exoU-基因型(P<0.05)。结论:本研究检出T3SS致病毒力基因以exoT+/exoY+/exoS+/exoU-型为主,不同毒力基因型组合的PA对常见抗生素的耐药性有差异。

多重耐药假单胞菌属细菌对碳青霉烯类和氨基糖苷类抗生素耐药的分子机制研究

【目的】探讨多重耐药假单胞菌属细菌对碳青霉烯类和氨基糖苷类抗生素耐药的分子机制。【方法】选2022年1月至12月于本院检验科收到的来自该医院不同病区的6株多重耐药恶臭假单胞菌和铜绿假单胞菌。采用Vitek-2compact全自动鉴定仪和16s rDNA序列引物鉴定并核实细菌种属。E-test药敏试条法测定菌株对临床抗生素抑菌浓度。PCR扩增和序列对比明确氨基糖苷类和碳青霉烯酶类甲基化酶的基因亚型。【结果】6株假单胞菌属细菌中有4株铜绿假单胞菌、2株恶臭假单胞菌,均为Carba NP阳性,且产碳青霉烯酶;菌株对青霉素类、碳青霉烯类、氨基糖苷类、头孢菌素类、四环素类抗生素均显示为耐药性。6株菌株中,仅SY456为氨曲类耐药,最小抑菌浓度值(MIC值)为92μg/mL,而其他菌株均为氨曲类耐药中介或敏感。Carba NP法筛查出6株菌均产耐药B类碳青酶烯酶。经与overlap PCR序列比对显示,6株菌株携带armA基因和blaIMP-45基因,3株恶臭假单胞菌SY47、SY153和SY434之间的脉冲场凝胶电泳(PFGE)带型差异显著。【结论】本院存在同时携带armA和blaIMP-45的多重耐药恶臭假单胞菌和铜绿假单胞菌,临床医师需根据耐药菌株类型调整用药策略。

世界卫生组织《2024年细菌类重点病原体目录》

抗菌药物耐药性是全球公共卫生面临的主要挑战之一,2019年约有495万相关死亡病例,高于低收入和中等收入国家的死亡人数比例。抗菌药物耐药性在很大程度上由错误和过度使用抗微生物药物造成。世界卫生组织(WHO)于2024年5月17日发布了其更新的《2024年细菌类重点病原体目录》。

基于网络药理学及细胞实验探讨扶正透邪解毒方治疗铜绿假单胞菌肺炎作用机制

目的:筛选扶正透邪解毒方抑制铜绿假单胞菌肺炎的中药活性化合物,并通过体外细胞实验研究扶正透邪解毒方治疗铜绿假单胞菌肺炎的作用机制。方法:在GeneCard、OMIM等数据库筛选铜绿假单胞菌肺炎疾病靶点;明确扶正透邪解毒方活性成分与铜绿假单胞菌肺炎的共同潜在靶标;利用String数据库和Cytoscape软件构建蛋白质-蛋白质相互作用(PPI)网络,利用DAVID数据库进行基因本体(GO)功能及京都基因与基因组百科全书(KEGG)通路富集分析;体外细胞实验中采用CD4^(+)T细胞与多重耐药铜绿假单胞菌共培养,以扶正透邪解毒方含药血清干预后,采用RT-PCR检测CD4^(+)T细胞中肿瘤蛋白P53(TP53)mRNA、丝氨酸/苏氨酸蛋白激酶1(AKT1)mRNA、白介素-6(IL-6)mRNA、白介素-10(IL-10)mRNA、白介素-1β(IL-1β)mRNA、肿瘤坏死因子-α(TNF-α)mRNA表达,采用Western blotting检测CD4^(+)T细胞中IL-6、TNF-α、AKT1、IL-1β、TP53、IL-10蛋白表达。结果:筛选出226种潜在活性成分及93个潜在靶点;TP53、AKT1、IL-6、IL-10、IL-1β、TNF-α等是扶正透邪解毒方治疗铜绿假单胞菌肺炎的关键靶点;扶正透邪解毒方主要通过炎症因子、趋化因子、免疫调控等信号通路来发挥治疗铜绿假单胞菌肺炎的作用。扶正透邪解毒方含药血清能降低CD4^(+)T细胞IL-6 mRNA、TNF-αmRNA、AKT1 mRNA、IL-1βmRNA表达(P<0.01),促进CD4+T细胞TP53 mRNA、IL-10 mRNA表达(P<0.01);扶正透邪解毒方含药血清能降低CD4^(+)T细胞IL-6、TNF-α、AKT1、IL-1β蛋白表达(P<0.05),促进CD4^(+)T细胞TP53、IL-10蛋白表达(P<0.05)。结论:扶正透邪解毒方可能是通过调控TNF-α、AKT1、TP53、IL-10等关键因子的表达抑制肺部感染而治疗铜绿假单胞菌肺炎。

河南某肿瘤医院临床分离菌分布及耐药性分析

目的:分析河南某肿瘤医院临床分离菌的分布及耐药性,为临床合理用药提供依据。方法:采用WHONET 5.6软件,分析2019年1月~2021年12月期间该院临床分离的所有细菌分布情况及耐药性。结果:共收集非重复菌株14769株,其中革兰阴性菌9863株(66.8%),革兰阳性菌4906株(33.2%)。标本以呼吸道(4295株,29.1%)、无菌体液(4093株,27.7%)和血液(2576株,17.4%)为主,科室分布以血液科(3134株,21.2%)、胃肠外科(1737株,11.8%)、肝胆外科(1703株,11.5%)、骨科(956株,6.5%)和重症医学病房(ICU,808株,5.5%)为主。耐甲氧西林金黄色葡萄球菌(MRSA)和耐甲氧西林凝固酶阴性葡萄球菌(MRCNS)3年的检出率依次为35.5%(175/493)、36.4%(176/483)、41.2%(206/500)和78.8%(246/312)、65.6%(231/352)、61.7%(227/368)。屎肠球菌对多数抗菌药物的耐药率均高于粪肠球菌。大肠埃希菌对碳青霉烯类仍高度敏感,3年的耐药率均低于5.0%。肺炎克雷伯菌对亚胺培南和美罗培南3年的耐药率呈下降趋势。铜绿假单胞菌对大多数抗菌药物的耐药率均低于鲍曼不动杆菌。ICU和血液科多重耐药菌的检出率较高。结论:该院2019年1月~2021年12月期间肿瘤患者临床分离菌株以革兰阴性杆菌为主,多重耐药菌除MRSA和CRPA有所上升外,其他均呈下降趋势,应继续加强医院感染防控,重视细菌耐药监测。

黏液型铜绿假单胞菌76株对常用抗菌药耐药性检测及毒力基因分布

目的探讨76株黏液型铜绿假单胞菌(PA)对常用抗菌药物耐药性检测及毒力基因分布。方法76株黏液型PA均分离于2017年1月至2021年12月浙江省宁海县中医医院住院的患者标本,全自动微生物分析仪鉴定菌株,采用纸片扩散法检测黏液型PA菌株对亚胺培南(IPM)、阿米卡星(AK)、妥布霉素(TOB)、庆大霉素(CN)、头孢吡肟(FEP)、哌拉西林/他唑巴坦(TZP)、左氧氟沙星(LEV)、头孢他啶(CAZ)、环丙沙星(CIP)和美罗培南(MEM)的敏感度,聚合酶链反应(PCR)法检测菌株Ⅲ型分泌系统(T3SS)效应蛋白基因exoS、exoT、exoU、exoY的分布。分析76株黏液型PA的来源及对常用抗菌药物耐药性分析、T3SS毒力基因的分布特征;比较ExoT^(+)/ExoY^(+)/ExoS^(+)/ExoU^(-)及ExoT^(+)/ExoY^(+)/ExoS^(-)/ExoU^(+)基因型与常用抗菌药物的关系。结果76株黏液型PA来源以痰液最多(68%);黏液型PA对AK、TOB、CN、TZP、LEV、CAZ的耐药率较低,对IPM、MEM、FEP和CIP的耐药率较高;黏液型PA在48 h对常见抗菌药物的耐药率为7%~14%,与24 h药敏结果(0~9%)差异有统计学意义(P<0.05);PA分离株中ExoT毒力基因的阳性率最高(100%),其次为ExoY(92%)、ExoS(70%),ExoU(26%)最低;T3SS毒力基因分型中,ExoT^(+)/ExoY^(+)/ExoS^(+)/ExoU^(-)(67%)和ExoT^(+)/ExoY^(+)/ExoS^(-)/ExoU^(+)占比最高(22%);ExoT^(+)/ExoY^(+)/ExoS^(+)/ExoU^(-)基因型对LEV、IPM、CIP的耐药率分别为37%、22%和41%,低于ExoT^(+)/ExoY^(+)/ExoS^(-)/ExoU^(+)基因型的100%、76%和100%(P<0.05);ExoT^(+)/ExoY^(+)/ExoS^(+)/ExoU^(-)基因型对FEP、TZP的耐药率为84%、78%,高于ExoT^(+)/ExoY^(+)/ExoS^(-)/ExoU^(+)基因型的35%、41%(P<0.05)。结论黏液型PA对碳青霉烯类抗菌药物IPM、MEM耐药率较高,T3SS毒力基因ExoT、ExoY在黏液型PA中普遍存在,不同毒力基因型组合与黏液型PA对常见抗菌药物的耐药率有关。

抗菌涂料抗菌性能测定方法的扩展与分析

为更好分析不同抗菌涂料对多种细菌的抗菌效果,先利用梯度稀释法确定四种细菌菌液的适宜测试浓度,消除菌液丰度的影响,然后按《抗菌涂料(漆膜)抗菌性测定法和抗菌效果》(GB/T 21866—2008)与《食品卫生微生物学检验菌落总数测定》(GB/T 4789.2—2016)等标准方法,测定了硅酸盐和竹炭两种类型抗菌涂料对大肠杆菌、金黄色葡萄球菌、肺炎克雷伯氏菌、铜绿假单胞菌的抗菌性能及其耐久性.结果表明,菌悬液制备过程中菌落的适宜计数稀释梯度均为10^(-5)和10^(-6);在24 h后两种抗菌涂料对4种细菌的抗菌率和抗菌耐久性分别达到99.35%和98.18%以上,对比非抗菌涂料,抗菌性能达到Ⅰ级标准.两种抗菌涂料均适用于医院、幼儿园等公共卫生条件要求较高的场所.

对饮用水中疑似铜绿假单胞菌的复核鉴定

铜绿假单胞菌是一种不允许在饮用水中检出的条件致病菌,本文对10株分离自饮用水样品的菌株进行了菌落形态观察及生化反应确证,并使用HK-MID细菌鉴定系统、VITEK 2全自动微生物鉴定系统和16S rRNA基因测序等多种手段进行进一步鉴定。鉴定结果表明,部分菌株并非铜绿假单胞菌,但均为条件致病菌,从食品安全的角度考虑,不应出现在饮用水、矿泉水中。本文除了探讨以往的研究普遍关注的检验方法的准确性与时效性,还对这些菌株进行了分析,为相关管理部门进行质量监督、制定标准提供了研究依据和借鉴。