ANAE染色和抗Thy1-SPA菌体花环双标记法的建立及其用于检测小鼠T淋巴细胞的研究

目的建立 ANAE染色联合抗 Thy1SPA菌体花环双标记法 ,并试图利用此法证明抗θ抗体对 ANAE阳性淋巴细胞的特异作用。方法利用双标记法对正常小鼠胸腺及脾细胞进行检测 ,观察抗 Thy1SPA抗体对 ANAE阳性细胞结合的专一性。结果双标记法中 ,抗 Thy1SPA菌体花环阳性同 ANAE阳性一样 ,能很好反映 6 15小鼠胸腺或脾脏 T细胞数 (P>0 .0 5 ) ;胸腺和脾脏细胞 Thy1+ - ANAE+ 率 ,即双标记阳性率略低于总的 ANAE+ 或总抗θ菌体花环 + 数 ,但 3者之间有明显的相关性。结论成功建立了 ANAE-抗 Thy1SPA菌体花环法 ,并首次用此法证明抗θ血清对

三种百合科植物抗真菌实验

采用百合科植物──大蒜、大葱、韭菜捣碎制成的膏体进行体外抗常见皮肤丝状菌的测定。以药物敏感试验法作为考核该植物的抑制皮肤浅部真菌的作用。结果表明，上述植物对常见皮肤真菌──红色癣菌和断发癣菌都有明显的作用，并将此作用与市售常用的抗真菌外用药进行了比较，提示这些植物药效成份的提取可作为一种新的治疗脚癣抗真菌的药物。

部分水稻杂交组合的抗稻瘟病性研究

:1 998～ 1 999年通过对苗期混合接种鉴定、抗菌系谱测定、田间自然诱发鉴定等不同方法 ,对部分水稻杂交组合进行了抗稻瘟病性研究。结果表明 ,虽然部分品种在苗期抗性鉴定时表现为抗性 ,但在抗菌系谱测定中所有品种都存在致病力不同 ,数量不等的侵染菌系 ,而田间自然诱发鉴定表明了在苗期抗性鉴定中表现抗性的品种成株期对穗颈瘟仍表现感病。为此 ,我们提出一定要注意品种的合理布局和搭配种植 ,避免在一个地区连续种植同一品种 ,以尽可能延长品种的使用年限。同时必须结合感病生育期的气候条件 。

减毒沙门菌介导防御素-3乳腺表达及对乳腺炎主要病原菌的抗菌效果

探讨减毒沙门菌介导的人HNP-3原前肽和前肽基因乳腺表达及其表达产物对牛乳腺炎主要病原菌的抗菌效果。构建人HNP-3原前肽和前肽基因的乳腺表达载体pEGFP-WAP-prepro-HNP-3和pEGFP-WAP-pro-HNP-3,电转化导入减毒沙门菌ΔcrpC78-1,构建重组菌ΔcrpC78-1（pEGFP-WAP-prepro-HNP-3）和ΔcrpC78-1（pEGFPWAP-pro-HNP-3）,重组菌分点注射到怀孕后20~22d母兔腹部乳腺分布区,收集兔分娩后不同时间的泌乳,Western-blot和荧光检测乳中HNP-3融合蛋白表达情况,ELISA检测乳中HNP-3融合蛋白的表达水平,常规方法分离乳腺炎主要病原菌,活菌计数法检测泌乳中HNP-3融合蛋白抑菌效果。结果显示,乳汁中分别在40 000和35 000处有prepro-HNP-3和pro-HNP-3融合蛋白阳性杂交信号,且在488nm处有黄绿色荧光;母兔在分娩后第5dHNP-3融合蛋白表达量最高,分别可达584μg/L和470μg/L,及至第25天表达量仍可达到277μg/L和213μg/L。活菌计数法结果表明分娩后第5d含prepro-HNP-3融合蛋白兔乳对牛乳腺炎主要病原菌大肠埃希菌、溶血巴氏杆菌、无乳链球菌、乳房链球菌、金黄色葡萄球菌和表皮葡萄球菌的抑菌率分别为92.0%、54.8%、46.4%、48.8%、86.9%和74.9%;含pro-HNP-3融合蛋白乳的抑菌率分别为84.9%、46.5%、40.6%、40.4%、71.1%和58.4%。结果表明,减毒沙门菌能够介导人HNP-3融合蛋白基因在乳腺上皮细胞中定位表达,表达的preproHNP-3和pro-HNP-3具有较强的生物学活性,对牛乳腺炎主要病原菌具有明显的抑制作用,为乳腺炎的生物防控及生物源性抗菌肽的研究提供了参考。

Noninvasive molecular analysis of Helicobacter pylori : Is it time for tailored first-line therapy?

The main problem of Helicobacter pylori(H. pylori) infection management is linked to antibiotic resistances. This phenomenon has grown in the last decade, inducing a dramatic decline in conventional regimen effectiveness. The causes of resistance are point mutations in bacterial DNA, which interfere with antibiotic mechanism of action, especially clarithromycin and levofloxacin. Therefore, international guidelines have recently discouraged their use in areas with a relevant resistance percentage, suggesting first-line schedules with expected high eradication rates, i.e., bismuth containing or non-bismuth quadruple therapies. These regimens require the daily assumption of a large number of tablets. Consequently, a complete adherence is expected only in subjects who may be motivated by the presence of major disorders. However, an incomplete adherence to antibiotic therapies may lead to resistance onset, since sub-inhibitory concentrations could stimulate the selection of resistant mutants. Of note, a recent meta-analysis suggests that susceptibility tests may be more useful for the choice of first than second-line or rescue treatment. Additionally, susceptibility guided therapy has been demonstrated to be highly effective and superior to empiric treatments by both meta-analyses and recent clinical studies. Conventional susceptibility test is represented by culture and antibiogram. However, the method is not available everywhere mainly for methodology-related factors and fails to detect hetero-resistances. Polymerase chain reaction(PCR)-based, culture-free techniques on gastric biopsy samples are accurate in finding even minimal traces of genotypic resistant strains and hetero-resistant status by the identification of specific point mutations. The need for an invasive endoscopic procedure has been the most important limit to their spread. A further step has, moreover, been the detection of point mutations in bacterial DNA fecal samples. Few studies on clarithromycin susceptibility have shown an overall high sensitivity and specificity when compared with culture or PCR on gastric biopsies. On these bases, two commercial tests are now available although they have shown some controversial findings. A novel PCR method showed a full concordance between tissue and stool results in a preliminary experience. In conclusion, despite poor validation, there is increasing evidence of a potential availability of noninvasive investigations able to detect H. pylori resistances to antibiotics. These kinds of analysis are currently at a very early phase of development and caution should be paid about their clinical application. Only further studies aimed to evaluate their sensitivity and specificity will afford novel data for solid considerations. Nevertheless, noninvasive molecular tests may improve patient compliance, time/cost of infection management and therapeutic outcome. Moreover, the potential risk of a future increase of resistance to quadruple regimens as a consequence of their use on large scale and incomplete patient adherence could be avoided.

Determination of the Content of Total Alkaloids in Different Solvent Extracts from Climbing Groundsel Herb Produced in Guizhou Province and Their Antibacterial Activity

[Objective] This study aimed to compare the contents of total alkaloids in different solvent extracts from climbing groundsel herb produced in Guizhou Province and their antibacterial activity. [Method] The content of total alkaloids was measured by UV spectrophotometry, and Oxford cups were used to investigate the antibacterial activity of each solvent extract. [Result] There was a good linear correlation between the absorbance measured by the UV spectrophotometer and the content of total al- kaloids within the concentration of 0.011 0-0.054 8 mg/ml at 207 nm, and the re- gression equation was Y=23.654X＋0.021, R=0. 999 7 and the average recovery rate was 99.2%. The contents of total alkaloids in 60% ethanol extract, 95% ethanol and water ethanol were 38.71, 52.25 and 60.50 mg/g, respectively. The 60% ethanol ex- tract had strong antibacterial activity against Streptococcus pneumoniae, Staphylo- coccus aureus and Escherichia coil; the water extractive had stronger antibacterial activity against Escherichia colr, 95% ethanol extract had weak antibacterial activity, with no inhibition of Pseudomonas aeruginosa. [Conclusion] This method is accurate, simple, with good repeatability, which can be used for the determination of alkaloids content of the climbing groundsel herb; there is no positive correlation between the content of total alkaloids in different solvent extracts of climbing groundsel herb and their antibacterial activity.

Determination of fluorinated quinolone antibacterials by ion chromatography with fluorescence detection

For preparing fluorinated quinolone antibiotic medicine locally used in stomatology, simultaneous determination of norfloxacin, ciprofloxacin, and enoxacin was carried out by multiphase ion chromatography with fluorescence detection. Quinolone antibiotics were separated by Dionex OmniPac PAX-500 column with an eluent of 15 mmol/L H2SO4 and 35% methanol （v/v） at a flow-rate of 1.0 ml/min and detected with fluorescence with excitation and emission wave lengths of 347 ran and 420 ran respectively. The detection limits （S/N=3） of norfloxacin, ciprofloxacin and enoxacin were 50, 105 and 80 ng/ml respectively. The relative standard deviations of retention time, peak area and peak height were less than 1.1% and good linear relationship resulted. The developed method was applied to pharmaceutical formulations and biological fluids.

Determination of Antimicrobial Activity of Methyl Cellulose Films Enriched with Some Essential Oils on Listeria innocua

Essential oils are effective antimicrobials on important some pathogenic bacteria and can be added packaging materials due to absorb various surfaces. In this study, the aim is to determinate of antimicrobial effects of methyl cellulose films with containing various proportions thyme, rosemary, coriander, basil and pimento berry oils （1%, 2%, 3% and 4% g/cc） against important bacteria as a surface contaminant Listeria innocua by in vitro. For this purpose, 1 cm diameter discs were cut from methyl cellulose films containing essential oils and inhibition zone diameter which were measured by the agar diffusion method. As the results of these analyses, all prepared active edible methyl cellulose films were antimicrobial effects on Listeria innocua. The most powerful antimicrobial effect achieved with methyl cellulose film including coriander oil on Listeria innocua. And also the weakest antimicrobial effect achieved with methyl cellulose film including rosemary oil on Listeria. All results were significant by statistically （p 〈 0.01） and all analyses were replicated in three times.