染

Wtp53 has obvious antiblastic effect towards Y79 cells, and the apoptosis rate of Y79 cells was the highest after transfection mediated by ultrasound-microbubble.

结论wtp53基因对RB瘤细胞具有较明显的抑制生长的作用；质粒+微泡+超声辐照，质粒+脂质体转染，以及质粒+超声辐照，均可促进Y79细胞的凋亡，但超声微泡介导的转染引起的凋亡率高于脂质体转染，而二者的凋亡率又明显高于质粒+超声转染。

Three RNAi target sites (named as Abil1, Abil2, Abil3) targeting the E3B1 (NCBI: NM-024397), an identified target sites and positive control GAPDH-A were selected. The pGenesil-1 eukaryotic expression vectors with the neoR mark and GFP green fluorescent mark were selected to construct E3B1 RNAi plasmid. The effect of RNAi targeting various sites on E3B1 genes expression was evaluated by testing the transfection efficiency with fluorescence microscope and western blot. The most effective siRNA in inhibiting E3B1 genes were screened. The most effective siRNA in inhibiting E3B1 genes screened and used to transfect the neuronal cells. After that, the inhibitor of axon growth was added and the axon growth was observed. Result: The siRNA expressing plasmids targeting the E3B1 were constructed successfully.

选择针对E3B1(NCBI： NM-024397)RNAi的3个靶位点Abil1、Abil2、Abil3和1个不针对任何mRNA的RNAi靶位点以及甘油醛－3-磷酸脱氢酶，用带有neoR选择标志和GFP绿色荧光标志的真核表达载体pGenesil-1构建E3B1 RNAi质粒，分别转染培养的神经元，在荧光显微镜下观测转染率，经G418筛选得到单一的转染细胞，并用Western blot法检测各转染组神经元E3B1蛋白的表达情况，选出具有最佳抑制效应的siRNA转染神经元，加入轴突生长抑制物，观测轴突生长情况。

Inaddition,we extract total RNA from cultural U251 cell of humanglioma,make coding gene extron amplification of TfR through ReverseTranscription-Polymerase Chain Reaction,construct expressionplasmid of pCDNA3.1-TfR after digestion of enzyme andconjunction,do transfection to U251 cell by liposome afteridentification,cause excess expression,detect the efficiency of transientexpression of TfR protein by pEGFPN1-TfR, sieve positive cell ofpCDNA3.1-TfR through G418, choose positive cell clone, detect theexpression of TfR in anteroposterior period through Western blot,FCM,cell immunochemistry and mRNA through RT-PCR, identify the effect of transfection.

另外，培养人脑胶质瘤U251细胞，进行细胞总RNA抽提，运用RT-PCR方法进行TfR基因的完整编码区外显子的扩增，经过酶切、连接后构建pCDNA3.1-TfR的表达质粒，鉴定正确后按脂质体转染方法转染人胶质瘤U251细胞，使其过量表达，通过pEGFPN1-TfR检测其瞬时转染的效率；用G418对稳定表达pCDNA3.1-TfR的细胞进行筛选，挑选阳性细胞克隆，运用Western blot、流式细胞检测技术和细胞免疫化学以及mRNA半定量的方法检测TfR在转染前后的表达水平，鉴定转染的效果。

The stable clones are further identified by RT-PCR and Western blot; 6 MTT assay is used to investigate the effect of ZNRD1 on the cell growth of cells (AGS, SGC7901, MKN28, NIH3T3, GES-1); 7 Soft agar assay is used to investigate the effect of ZNRD1 on the clonality of cells (AGS, MKN28); 8 Nude mice assay is used to investigate the effect of ZNRD1 on the cell growth of gastric cancer cells (AGS, MKN28); 9 Flow cytometry is used to investigate the effect of ZNRD1 on the cell cycle distribution of cells (AGS, MKN28, NIH3T3, GES-1); 10 Flow cytometry is used to investigate the effect of ZNRD1 on the cell apoptosis of cells (AGS, MKN28, NIH3T3); 11 MTT assay is used to investigate the effect of ZNRD1 on the drug sensitivity of cancer cells (SGC7901, SGC7901/VCR, HL-60, HL-60/VCR) in vitro; 12 SRCA is used to investigate the effect of ZNRD1 on the drug sensitivity of gastric cancer cells (SGC7901, SGC7901/VCR) in vivo; 13 Flow cytometry is used to investigate the effect of ZNRD1 on adriamycin accumulation of cancer cells (SGC7901, SGC7901/VCR, HL-60, HL-60/VCR); 14 Transmission electron microscope is used to investigate the effect of ZNRD1 on the sensitivity of SGC7901 cells towards drug-induced apoptosis; 15 Flow cytometry and DNA ladder assay are used to investigate the effect of ZNRD1 on the sensitivity of cells (SGC7901, SGC7901/VCR, HL-60/VCR) towards drug-induced apoptosis; 16 Microarray is used to investigate the profiling of ZNRD1-responsive genes in gastric cancer cells (AGS, MKN28, SGC7901, SGC7901/VCR); 17 RT-PCR and Western blot are used to identify the results of microarray; 18 Reporter gene assay is used to investigate the effect of ZNRD1 on the transcriptional activity of cyclin D1; 19 Reporter gene assay is used to investigate the effect of ZNRD1 on the transcriptional activity of MDR1; 20 Kinase assay is used to investigate the effect of ZNRD1 on the activity of cyclin E-CDK2 kinase; 21 The antisensenucleic acids of p21 is used to inhibit the expression of p21, and flow cytometry is used to investigate the effect of p21 on ZNRD1-induced cell cycle arrest in gastric cancer cells; 22 The antisensenucleic acids of p27 is used to inhibit the expression of p27, and flow cytometry is used to investigate the effect of p27 on ZNRD1-induced cell cycle arrest in gastric cancer cells; 23 Liposome is used to up-regulate the expression of Skp2, and flow cytometry is used to investigate the effect of Skp2 on ZNRD1-induced cell cycle arrest in gastric cancer cells; 24 Western blot is used to investigate the effect of ZNRD1 on the stability of Skp2 and p27 in gastric cancer cells; 25 MVD assay is used to investigate the effect of ZNRD1 on the angiopoietic activity of gastric cancer cells; 26 ELISA is used to investigate the effect of ZNRD1 on the expression of VEGF165 in gastric cancer cells; 27 The roles of DARPP-32 in MDR of gastric cancer cells are investigated using gene transfection, MTT assay, SRCA, flow cytometry and DNA ladder assay.

应用杂交瘤技术制备ZNRD1的首个单克隆抗体；2）利用RT-PCR、Western blot和免疫组化检测ZNRD1在胃癌组织、胃炎组织、正常胃上皮组织、胃癌细胞和正常胃组织上皮细胞中的表达；3）构建ZNRD1的小干扰RNA载体，并测序鉴定；4）利用脂质体将ZNRD1的真核表达载体及其空载体转染胃癌细胞（AGS、SGC7901、MKN28）和小鼠成纤维细胞（NIH3T3），G418筛选后进行鉴定；5）利用脂质体将ZNRD1的小干扰RNA载体及其空载体转染药敏胃癌细胞（SGC7901）、正常胃组织上皮细胞（GES-1）、对长春新碱耐药的胃癌细胞（SGC7901/VCR）、药敏白血病细胞（HL-60）、对长春新碱耐药的白血病细胞（HL-60/VCR），G418筛选后进行鉴定；6）利用MTT实验检测ZNRD1高/低表达对细胞（AGS、SGC7901、MKN28、NIH3T3、GES-1）生长的影响；7）通过软琼脂克隆形成实验检测上调ZNRD1对AGS、MKN28细胞克隆形成能力的影响；8）通过裸鼠成瘤实验检测上调ZNRD1对AGS、MKN28细胞体内成瘤性的影响；9）通过流式细胞仪分析ZNRD1高/低表达对细胞（AGS、MKN28、NIH3T3、GES-1）的细胞周期的影响；10）通过流式细胞仪分析上调ZNRD1对细胞（AGS、MKN28、NIH3T3）的凋亡的影响；11）通过MTT实验检测ZNRD1高/低表达对细胞（SGC7901、SGC7901/VCR、HL-60、HL-60/VCR）体外药物敏感性的影响；12）通过肾包膜下移植法检测ZNRD1高/低表达对细胞（SGC7901、SGC7901/VCR）体内药物敏感性的影响；13）通过流式细胞仪分析ZNRD1高/低表达对细胞（SGC7901、SGC7901/VCR、HL-60、HL-60/VCR）内阿霉素蓄积和泵出的影响；14）通过透射电镜检测上调ZNRD1对SGC7901细胞凋亡敏感性的影响；15）通过流式细胞仪和DNA梯度试验检测ZNRD1高/低表达对细胞（SGC7901、SGC7901/VCR、HL-60）凋亡敏感性的影响；16）通过基因芯片检测ZNRD1高/低表达对胃癌细胞内基因表达谱的影响；17）利用RT-PCR、Western blot对基因芯片的结果进行鉴定；18）利用报告基因实验检测ZNRD1对cyclin D1的启动子活性的调节作用；19）利用报告基因实验检测ZNRD1高/低表达对MDR1的启动子活性的调节作用；20）利用激酶试验检测ZNRD1对cyclin E-CDK2 激酶活力的影响；21）利用反义核酸技术抑制p21的表达；通过流式细胞仪检测抑制p21对ZNRD1介导的细胞周期阻滞的影响；22）利用反义核酸技术抑制p27的表达；通过流式细胞仪检测抑制p27对ZNRD1介导的细胞周期阻滞的影响；23）利用脂质体转染法上调Skp2的表达；通过流式细胞仪检测上调Skp2对ZNRD1介导的细胞周期阻滞的影响；24）利用Western blot检测ZNRD1对p27和Skp2的蛋白稳定性的影响；25）利用微血管密度实验检测ZNRD1对AGS、MKN28细胞裸鼠移植瘤微血管形成的影响；26）利用ELISA检测ZNRD1对AGS、MKN28细胞培养上清和移植瘤匀浆中VEGF165含量的影响；27）利用脂质体转染法、MTT实验、肾包膜下移植法、流式细胞仪和DNA梯度试验检测新耐药相关分子DARPP-32对细胞（SGC7901、SGC7901/VCR、对阿霉素耐药的胃癌细胞SGC7901/ADR）多药耐药表型的影响；利用脂质体转染法和MTT实验检测下调ZNRD1对DARPP-32介导的胃癌多药耐药的调控作用。

1,After transfected the mutated mtDNA of colorectal carcinoma,the mtDNA D-loop region of the transfected cells displays new mutation points.2,The external source pieces of the mutated mtDNA can integrate to nuclear genome after transfection.3,There's no differences in apoptosis between combinations after transfected the mutation of mtDNA in NIH3T3 and LST cells.4,The mutated mtDNA may affect the action mechanism of occurrence and development in colorectal carcinoma through affecting its mtDNA mutation or integrating exogenetic mtDNA to its nuclear which may cause the abnormal expression of oncogene or anti-oncogene.

（1）转染突变的大肠癌细胞mtDNA后转染细胞的mtDNA均可发生多处的突变位点。（2）通过转染后突变的外源性的mtDNA可以整合到核基因组内。（3）突变的mtDNA转染LST细胞及NIH3T3细胞后，不影响转染细胞的凋亡改变。（4）mtDNA的突变可能通过影响体细胞mtDNA的突变和通过外源性mtDNA在核内的整合从而影响癌基因或抑癌基因的表达异常，从而参与肿瘤的发生发展。

Single factor method were compared to the concentration of dyes, dye bath pH, temperature, Dyeing time to do a series of gradient experiment, the two groups reached on experiments with suitable for dyeing process conditions,such as the best dye pH, suitable dye concentration, Dyeing appropriate time,temperature values of humic acid and fulvic acid were dyeing wool and silk, and which the general situation has deep understanding.

采用单因素对比法分别对染料的浓度、染液pH值、染色温度、染色时间等做了系列梯度实验，得出了两组上染实验的最佳染液pH值、染料合适浓度、适当染色时间、适宜上染温度等工艺条件值，对腐植酸和黄腐酸分别上染羊毛线及真丝绸的大体情况有了很深的了解。

In this paper, The influence of heat set and dyeing on the structure of PTT fiber was investigated by FT-IR, DSC, critical dissolution time. The influence of heat set on the dyeability of PTT fiber was discussed, too. Meanwhile, many experiments on the dyeing performance of PET fiber were carried out. The effect of different dyeing temperature on the uptake and visual color depth value and dyeing characteristics of different dyes with temperature increasing were discussed, and the influence of temperature on the dyeing depth were also evaluated when dyeing for PTT and PET fiber in the same dye bath condition. Especially the kinetics and thermodynamics of dyeing with disperse dyes were mainly studied.

目前，我国对PTT染色性能方面研究报道还少，本文用傅立叶红外光谱、差热分析、临界溶解时间等分析方法研究了热定型和染色对PTT纤维结构的影响，探讨了热定型对PTT纤维可染性的影响，同时进行了多项PTT纤维的基本染色性能试验，探讨了不同染色温度对上染百分率及表观色深值的影响、不同染料的升温上染特性、不同染料的移染性能和染深性能以及PTT和PET纤维同浴染色时染色温度对它们的染色深度的影响。

On the raw fur process, Zhonghui can tan the fur of mink, blue fox, raccoon, rabbit, sheep goat and so on. On the dyeing, Zhonghui can dye single color, double color, mixed color, snowtop, print pattern, tie-dye, and so on. On Bleaching and dyeing, we can golden mink, bleach raccoon, and other kinds of products.

原皮加工方面，中辉鞣制水貂、蓝狐、貉子、兔皮、绵羊皮、山羊皮等皮种；染制单色、双色、多色以及草上霜、印花、扎花、吊染、型染等各种产品；漂染包括水貂漂金、貉子漂金、漂白以及其他漂染产品。

Our company owns factories of stained wash water for Knitted pieces and denim wash water for jeans, which enables it to offer kinds of external processing businesses, such as direct dyeing, acid dyeing, hanging dyeing, Knot dyeing, fermentation washing, etc. With its excellent work and unique style, the company's solid-color fastness has reached the standards of International Laboratory STS export testing.

公司属下针织件染洗水厂、牛仔洗水厂，为客户提供直接染、酸性染、碧纹染、吊染、扎染、酵洗、砂洗、雪洗、酵磨洗、转进洗、石磨、炒雪花、手工扫喷等对外加工洗染业务，洗水效果好，人衣合一，风格独特，其固色牢度已达到国际化验所STS出口测试标准。

The transient transfection efficiency was measured by flow cytometry. The stable transfected cells were screened by G418. Results Electrotransfection had the highest transient transfection efficiency (56.8%), with the most masc clones of stably transfection and the least time taken to form big masc clones (20 days). Cationic polymer mediated transient transfection had the lowest efficiency (1.7%), with the least masc clones and the longest time taken to form the big masc clones (30 days). Cationic liposome transfection had a transient transfection efficiency of 39.9% and took 25 days to form the big masc clones.

结果 电转染有最高的瞬时转染效率(56.8%)，稳定转染得到的阳性克隆最多，且从细胞筛选到扩增至得到阳性大克隆所用的时间也最少（约20d）；用阳离子聚合物瞬时转染率低(1.7%)，稳定筛选同样可以得到阳性大克隆，但是数量很少且从细胞筛选到扩增至阳性大克隆所用的时间也最多（约30d）；阳离子脂质体转染效果居中，瞬时转染率为39.9%，从细胞筛选到扩增至得到阳性大克隆所用的时间约25d。

Since historical times,England ,where the early inhabitants were Celts, has been conquered three times .

从有历史以来，英国，在此地早期居住的是凯尔特人，已经被征服了三次。

Bluetooth OBEX File Transfer Enables the sending and receiving of files on your phone via Bluetooth.

蓝牙OBEX文件移动允许经过蓝牙传送和接受文件。。。。