上皮细胞

Results:The results show that the survival rate of mammary epithelial cells were decreasing gradually with the increasing of the time of TGF-β1 treatment, and the survival rates of 24 h and 48 h groups were significantly lower than that of control group (p.05); the LDH activity of 6 h、12 h、24 h and 48 h groups were significantly higher than that of control group (p.05); the DNA degradation arisesed in 12 h, 24 h, and 48 h groups' mammary epithelial cells; the activity of Caspase-3 reach to a peak at 24 h, which was significantly higher than that of control group (p.05); the acinus of epithelial cells appears to break off and collapse with the increasing of treatment time, but the leydig structure keeps integrate;...

结果表明：1、5、10 ng/mL的TGF-β1均能抑制乳腺上皮细胞的增殖，且随浓度增加抑制作用加大，呈现剂量依赖性，其中10 ng/mL TGF-β1组与对照组差异显著（P.05）；10 ng/mL TGF-β1作用于乳腺上皮细胞24 h，随作用时间延长细胞凋亡率逐渐增加，6 h、12 h和24 h组与对照组相比差异极显著（P.01）；LDH活力也不断升高，6 h、12 h和24 h组与对照组差异显著（P.05）；DNA随 TGF-β1作用时间的延长发生不同程度降解；TGF-β1作用2 h~6 h细胞大量表达HSP70，6 h达到高峰，而后下降，2~12 h的表达量均显著高于对照组（P.05）。结论：TGF-β1可以抑制奶牛乳腺上皮细胞的增殖，具有浓度依赖性。10 ng/mLTGF-β1。。。

Was presented in 12 cases (24%) by bacterial isolation. By in-situ hybridization, PCV2 signals were chiefly distributed in interstitial and necrotic lesions, and the positive signals could be found in macrophages-like cells and necrotic debris. PRRSV signals were mainly located at interstitial lesions and alveolar wall, and the positive signals could be found in macrophages-like cells and epithelial cells of alveoli. Swine influenza virus could be found in interstitial and necrotizing lesions, and the signals could be found in macrophages-like cells, and epithelial cells of terminal bronchiole and alveoli.

另运用原位杂交法对PCV2、PRRSV及猪流行性感冒病毒进行检测，结果发现PCV2主要分布於PNP的间质及坏死病灶区，并能於肺泡及终末细支气管上皮细胞、巨噬细胞与坏死细胞碎片皆可见有病毒核酸；PRRSV则多分布於间质病灶区及肺泡壁处，能於巨噬细胞与肺泡上皮细胞见到病毒核酸；SIV则存在於间质及坏死病灶区，且能於肺泡及终末细支气管上皮细胞与巨噬细胞见有病毒核酸。

Quantitative chromatics analysis of large bowel tumorous adenoma and adenocarci-noma was made by the methods of computer image processing.

本文通过对大肠腺瘤和腺癌组织进行计算机图像定量分析，表明大肠腺瘤和腺癌的腺上皮细胞以正常杯状细胞和柱状细胞的色度学特征分布为基准可划分为两大类：杯状细胞性腺上皮和柱状细胞性腺上皮。

Results ERα immunoreactivity was detected in the nuclei of epithelial cells lining lobules and ducts.The positive cells located in the inner layer of the two epithelial layers in the lobules and intralobular ducts,but in the interlobular ducts the positive cells located in the outer layer.

结果：ERα在小叶上皮细胞和导管上皮细胞的细胞核内检测到，在小叶和小叶内导管分布于上皮细胞的内层，在小叶间导管分布于上皮细胞的外层。

A portion of ductal cells and individual serous acinic cells of the minor salivary gland also expressed Fas not only in membranous but also in cytoplasmic pattern.

结果发现，在正常鼻咽粘膜表面及隐窝的假复层纤毛柱状上皮细胞胞膜、部分小涎腺的导管上皮和个别浆液性腺泡上皮细胞的胞膜和胞浆Fas阳性表达；鳞状化生上皮的表层细胞胞膜有Fas表达，但基底细胞未见Fas表达；浸润的淋巴细胞也不表达Fas.45.9%（17/37）的鼻咽非角化性癌Fas阳性，但只有不足10%的癌细胞胞膜微弱表达Fas。

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。

PART 2:The molecular mechanisms of Nontypeable Haemophilus influenzae induced inflammatory response in respiratory epithelial cells Objective:Nontypeable Haemophilus influenzaewas traditionally considered as an extracellular pathogen in respiratory infections.In this study,we established a model of infected cells to confirm NTHI was also a facultatively intracellular pathogen.Mitogen activated protein kinasesare the important signal transduction pathway in mediating inflammatory response induced by NTHI in respiratory epithelium.In this study,we elucidate the impact of NTHI,and its SCF and EP on the expression of MAPK and subsequent proinflammatory mediators.

第二部分：NTHI致呼吸道上皮细胞炎症反应的分子机制研究目的：通过建立呼吸道上皮细胞株的体外NTHI感染模型，确认NTHI在细胞内的存活，观察NTHI及其组分(可溶性胞质成分SCF、外膜蛋白EP)对呼吸道上皮细胞MAPK信号传导通路和后续的前炎症因子表达的影响，并应用特异性ERK-MAPK抑制剂U0126、p38 MAPK抑制剂SB203580特异性阻断下游MAPK信号传导通路，观察它们对呼吸道上皮细胞分泌前炎症因子表达的影响。

The adhesion test showed that small intestinal epithelium cells of 30-35-day-old postweaning piglets with both genotype M307~ and M307~ could adhere to the standard E.coli strain express F18ab fimbriae, the recombinant E.coli express F18ac rE.coli1534 and the recombinant pnirBMisL-fedF E.coli displaying FedF subunit on the surface of E.coli, but small intestinal epithelium cells with genotype M307~ and 3-day-old piglet could not adhere to the three kinds of bacteria descried above, the latter with good adherence capability of 987P fimbrial E.coli.

9仔猪小肠上皮细胞的黏附试验结果显示，30～35日龄M307~基因型和M307~基因型断奶仔猪的小肠上皮细胞均能与表达F18ab菌毛标准菌株107／86、诱导表达F18ac菌毛的重组大肠杆菌rE.coli1534及诱导菌体表面展示表达F18黏附亚单位FedF的重组大肠杆菌pnirBMisL-fedF发生黏附作用，而同日龄M307~基因型断奶仔猪的小肠上皮细胞均不能与上述三株大肠杆菌发生黏附作用。3日龄易感仔猪小肠上皮细胞也不能黏附上述三株大肠杆菌，但可以很好地黏附表达于987P菌毛的大肠杆菌。

Primary culture of goat mammary gland cells could be derived by outgrowth of migrating cells from fragment of tissue. Fibroblast and epithelial cells could be pured according to their different sensibility to trypsin. Complex cultures treated by 0.25% trypsin in Hanks' for 5~7min at 37℃, the dispersed cells were mainly fibroblasts,then the rest treated by 0.15% trypsin-0.02% EDTA in Hanks'for 5~8min at 37℃, the majority cells harvested were mammary epithelial cells.

用组织块培养法可获得良好的山羊乳腺细胞原代培养物；培养的山羊乳腺成纤维细胞比上皮细胞对胰蛋白酶更敏感，据此可在传代过程中将二者分离纯化，获得纯细胞系；混合培养物先用0.25%胰蛋白酶在37℃消化5~7min 所收集的细胞主要为成纤维细胞；再加0.15%胰蛋白酶-0.02鞹A 消化液在37℃继续消化5~6min 所回收的细胞绝大多数为上皮细胞，经过2~3 代，即可得到纯化的乳腺上皮细胞系。

Primary culture of goat mammary gland cells could be derived by outgrowth of migrating cells from fragment of tissue. Fibroblast and epithelial cells could be pured according to their different sensibility to trypsin. Complex cultures treated by 0.25% trypsin in Hanks' for 5~7min at 37℃, the dispersed cells were mainly fibroblasts,then the rest treated by 0.15% trypsin-0.02% EDTA in Hanks'for 5~8min at 37℃, the majority cells harvested were mammary epithelial cells.

用组织块培养法可获得良好的山羊乳腺细胞原代培养物；培养的山羊乳腺成纤维细胞比上皮细胞对胰蛋白酶更敏感，据此可在传代过程中将二者分离纯化，获得纯细胞系；混合培养物先用0.25%胰蛋白酶在37℃消化5~7min 所收集的细胞主要为成纤维细胞；再加0.15%胰蛋白酶-0.02%EDTA 消化液在37℃继续消化5~6min 所回收的细胞绝大多数为上皮细胞，经过2~3 代，即可得到纯化的乳腺上皮细胞系。

It has been put forward that there exists single Ball point and double Ball points on the symmetrical connecting-rod curves of equilateral mechanisms.

从鲍尔点的形成原理出发，分析对称连杆曲线上鲍尔点的产生条件，提出等边机构的对称连杆曲线上有单鲍尔点和双鲍尔点。

The factory affiliated to the Group primarily manufactures multiple-purpose pincers, baking kits, knives, scissors, kitchenware, gardening tools and beauty care kits as well as other hardware tools, the annual production value of which reaches US$ 30 million dollars.

集团所属工厂主要生产多用钳、烤具、刀具、剪刀、厨具、花园工具、美容套等五金产品，年生产总值3000万美元，产品价廉物美、选料上乘、质量保证，深受国内外客户的青睐