应用噬菌体控制假单胞菌的研究进展

假单胞菌属细菌中的部分菌株是动植物和人类的致病菌，也是食品的腐败菌，会导致动植物和人类的高死亡率和严重的食品安全事故。噬菌体作为一种细菌性病毒，特异性高、自我增殖快且无副作用。这些特性使其成为了解决细菌性问题的新思路和新方法。本文就利用噬菌体防控假单胞菌属致病菌和腐败菌在食品、水产、植物、医疗等方面的研究和应用进行综述，以便为后期的深入研究提供一定的参考。     

Two-dimensional gel electrophoresis (2D-GE) image analysis based on CellProfiler: Pseudomonas aeruginosa AG1 as model

Two-dimensional gel electrophoresis (2D-GE) is an indispensable technique for the study of proteomes of biological systems, providing an assessment of changes in protein abundance under various experimental conditions. However, due to the complexity of 2D-GE gels, there is no systematic, automatic, and reproducible protocol for image analysis and specific implementations are required for each context. In addition, practically all available solutions are commercial, which implies high cost and little flexibility to modulate the parameters of the algorithms. Using the bacterial strain, Pseudomonas aeruginosaAG1 as a model, we obtained images from 2D-GE of periplasmic protein profiles when the strain was exposed to multiple conditions, including antibiotics. Then, we proceeded to implement and evaluate an image analysis protocol with open-source software, CellProfiler. First, a preprocessing step included a bUnwarpJ-Image pipeline for aligning 2D-GE images. Then, using CellProfiler, we standardized two pipelines for spots identification. Total spots recognition was achieved using segmentation by intensity, whose performance was evaluated when compared with a reference protocol. In a second pipeline with the same program, differential identification of spots was addressed when comparing pairs of protein profiles. Due to the characteristics of the programs used, our workflow can automatically analyze a large number of images and it is parallelizable, which is an advantage with respect to other implementations. Finally, we compared six experimental conditions of bacterial strain in the presence or absence of antibiotics, determining protein profiles relationships by applying clustering algorithms PCA (Principal Components Analysis) and HC (Hierarchical Clustering).     

铜绿假单胞菌注射液对肺癌A549细胞自噬及PI3K/Akt通路的影响

目的探究铜绿假单胞菌注射液(PA-MSHA)对肺癌A549细胞自噬及磷脂酰肌醇-3激酶(PI3K)/蛋白激酶B(Akt)通路的影响。方法体外培养人肺癌A549细胞,随机分为空白对照组、LY294002组(加入20μmol/L LY294002),PA-MSHA干预组(加入0.5×10^9/mL、1.0×10^9/mL、2.0×10^9/mL PA-MSHA)。干预培养48 h后,MTT法检测各组细胞活性,平板克隆形成实验检测各组细胞增殖能力,透射电子显微镜观察各组细胞自噬情况,Western blot检测各组细胞中自噬相关蛋白p62、LC3Ⅰ、LC3Ⅱ及PI3K/Akt通路相关蛋白PI3K、Akt、p-PI3K、p-Akt表达。结果与空白对照组相比,LY294002组、0.5×10^9/mL、1.0×10^9/mL、2.0×10^9/mL PA-MSHA组细胞增殖抑制率、LC3Ⅱ/LC3Ⅰ蛋白表达显著升高(P<0.05),自噬体个数增加,细胞克隆形成率、p62、p-PI3K/PI3K、p-Akt/Akt蛋白表达显著降低(P<0.05);与LY294002组相比,0.5×10^9/mL、1.0×10^9/mL PA-MSHA组细胞增殖抑制率、LC3Ⅱ/LC3Ⅰ蛋白表达显著降低(P<0.05),自噬体个数减少,细胞克隆形成率、p62、p-PI3K/PI3K、p-Akt/Akt蛋白表达显著升高(P<0.05);与0.5×10^9/mL、1.0×10^9/mL PA-MSHA组相比,2.0×10^9/mL PA-MSHA组细胞增殖抑制率、LC3Ⅱ/LC3Ⅰ蛋白表达显著升高(P<0.05),自噬体个数增加,细胞克隆形成率、p62、p-PI3K/PI3K、p-Akt/Akt蛋白表达显著降低(P<0.05)。结论PA-MSHA可能通过调控PI3K/Akt信号通路,促进肺癌A549细胞自噬,抑制细胞增殖。     

免疫调节剂对亚临床型生殖器疱疹豚鼠模型排毒及IL-2和IL-4水平的影响

目的观察匹多莫德联合铜绿假单胞菌注射液治疗亚临床型生殖器疱疹(genital herpes,GH)豚鼠模型的排毒及机体免疫的情况,借以揭示人类亚临床型GH患者联合服用免疫调节剂后的疗效。方法以人单纯疱疹病毒(HSV)-2Sav感染从未感染过HSV的雌性中华彩豚,建成GH豚鼠模型,待其症状好转后,通过聚合酶链反应(PCR)检测豚鼠外生殖器及生殖道拭子HSV-DNA的阳性率,酶联免疫吸附试验(ELISA)测出其血清白细胞介素(IL)-2、IL-4的浓度,再分别与空白对照组及用药组比较。结果亚临床型GH豚鼠模型仍有HSV排泌,且其IL-2浓度显著低于空白对照组(P0.01),而IL-4浓度高于空白对照组(P0.05);用药组的豚鼠HSV-DNA阳性率明显减少,而其血清IL-2浓度高于亚临床GH豚鼠模型组,IL-4的水平低于亚临床GH豚鼠模型组,差异有统计学意义(P0.05)。结论亚临床型GH豚鼠模型确实存在HSV排泌,同时其血清中细胞因子的分泌也发生了异常,匹多莫德联合铜绿假单胞菌注射液治疗后不仅能减少HSV排泌,还能改善机体免疫失衡状态。     

常规处理方案联合铜绿假单胞菌制剂治疗乳腺癌疗效的Meta分析

目的:系统性评价铜绿假单胞菌制剂应用于乳腺癌治疗中的效果。方法:检索国内外数据库,检索时间为各数据库建库至2014年11月。纳入在乳腺癌治疗中应用铜绿假单胞菌制剂的随机对照试验(RCT),筛选文献、提取数据和文献质量评价后,用Rev Man 5.1.2软件进行Meta分析。结果:共纳入9篇RCT研究,Meta分析结果显示,常规处理方案联合铜绿假单胞菌制剂明显升高患者完全缓解率(OR=2.22)、总有效率(OR=6.73)、患者Karnofsky评分(MD=9.79)及IL-2水平(SMD=1.56),明显降低患者疾病稳定率(OR=0.36)、疾病进展率(OR=0.18)以及缩短患者术后积液消失时间(MD=-7.52)。结论:联合铜绿假单胞菌制剂可明显提高乳腺癌患者放化疗效果,但该结论仍需要高质量的研究进一步证实。