胞质的

Drug sensitive test and three-dimensional test220 strains of Pa were isolated from hospitalized patients between 2003 and 2007. K-B method was used to tested the susceptibility of 10 different antibiotics. IRPa was screened by testing the minimal inhibitory concentration of imipemem by using agar diluiion method.The susceptibility of these IRPa to the antibiotics was analysised. Three-dimensional test was used to identify the different kinds of beta lactamases from 220 strains of Pa.2.Carbarpenems hydrolytic enzyme genes and oprD2 gene were detectedamong the selected IRPa strains, PCR method was performed to detect carbapenemase genes which included GES、KPC、SPM、VIM、IMP、GIM gene and the oprD2 gene;Multiplex PCR were used to detect OXA genes and plasmid-mediated AmpC beta lactamase genes; The expression of the chromosomal AmpC beta lactamases and oprD2 genes in IRPa strains were analyzed by Real-time PCR.3.Identification and characterization of integronsIntegrase gene was detected by PCR, and the classification of integrons was performed by using restriction fragment length polymorphism.PCR was performed to detect the qacE△1-sull gene,and the gene cassetes which are located at variable region of integrons in the strains were detected to be positive.

方法1、药敏实验和三维实验收集2003～2007年临床分离的220株Pa，对这些菌株采用K-B法测定10种临床常用抗生素的药敏情况，同时采用琼脂稀释法检测亚胺培南的最低抑菌浓度(Minimal inhibitory concentration，MIC)，筛选出对亚胺培南耐药的铜绿假单胞菌，并分析其对其它抗生素的药物敏感率；采用三维实验的方法分析220株Pa产β内酰胺酶的类型。2、碳青霉烯类水解酶和oprD2蛋白的检测针对鉴定的IRPa菌株，采用普通PCR方法检测具有碳青霉烯水解作用的β内酰胺酶耐药基因(GES、KPC、SPM、VIM、IMP、GIM基因)和oprD2基因，采用多重PCR的方法检测OXA型基因和质粒携带的AmpC酶基因，用荧光定量RT-PCR方法检测oprD2蛋白基因表达情况；同时对产AmpC酶的Pa(25株，含IMP耐药和敏感株)用RT-PCR方法检测AmpC酶基因的表达量情况。3。

Results:①The amount of human colon carcinoma cell line SW480 treated by quercetin decreased. The morphology of partial SW480 cells was shrunk volume, integrated cell membrane, condensed cytoplasm, pyknotic chromatin, nuclear fragmentation. Apoptotic Corpuscles were found by electron microscope.②MTT colorimetric assay showed quercetin inhibited the growth of human colon carcinoma cell line SW480 in a time- and dose-dependent manner when the concentration of quercetin was 30、60、90μmol/L.③Flow cytometry analysis showed the cell cycle of SW480 cell was restricted in G1/S. G0/G1 phase rate increased and S phase rate decreased with increasing concentration of quercetin and time lasting.④ Zymogram analysis assay showed the secretion of matrix metalloproteinases in human colon carcinoma cell line SW480 treated by quercetin decreased. With increasing concentration of quercetin, the secretion of MMP-2 and MMP-9 decreased.⑤Immunohistochemistry method demonstrated the position expression of Cathepsin-D in SW480 cell was suppressed by quercetin in a time- and dose-dependent manner.

研究结果：经槲皮素处理的人结肠癌SW480细胞数量减少，部分细胞体积缩小，细胞膜完整，胞浆浓缩，核染色质固缩，细胞核碎裂，形成凋亡小体；MTT法检测显示当作用浓度为30μmol/L~90μmol/L时，槲皮素对人结肠癌SW480细胞的生长有抑制作用，其抑制作用随着作用浓度的增加和作用时间的延长而增强；流式细胞学发现槲皮素主要作用于人结肠癌SW480细胞周期的G1/S期，大部分细胞被阻断于S期，随药物浓度的升高和作用时间的延长，G0/G1期细胞比例逐渐增加，S期细胞比例逐渐减少；酶谱分析法检测显示不同浓度的槲皮素能够抑制人结肠癌SW480细胞分泌MMP-2及MMP-9，随浓度的升高，MMP-2及MMP-9的分泌量减少；免疫组织化学法显示不同浓度的槲皮素处理人结肠癌SW480细胞后，Cathepsin-D的表达随药物浓度的升高和作用时间的延长而降低。

I didn't watch TV last night, because it .

昨晚我没有看电视，因为电视机坏了。

Since this year, in a lot of villages of Beijing, TV of elevator liquid crystal was removed.

今年以来，在北京的很多小区里，电梯液晶电视被撤了下来。