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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。

It is showed that the mechanism of XSTM was association with inhibiting expression of cell adhesion molecules of endothelium and inflammatory cell infiltrating into vessel wall, promoting fibrinolytic activity,inhibiting platelet adhesion and aggregation, regulating angiotasis, protecting the normal endothelium structure and function, inhibiting collagen hyperplasia and maintaining elasticity of vascular wall.

实验结果表明:消栓通脉颗粒剂具有显著抑制细胞黏附分子——P-选择素的表达,抑制炎性细胞的浸润,促进纤溶活性,抑制血小板黏附、聚集,调节血管张力,保护血管内皮细胞功能,抑制胶原纤维增生,维持静脉壁弹性等治疗作用。

The in vitro effects of Gln49-PLA_2 were examined using the chick biventer cervicis nerve muscle preparation. Gln49-PLA_2 caused a dosedependent blockade of neuromuscular transmission. The time required to cause 50% blockade was about 150 min at the highest concentration 100μg/ml. This inhibition appeared to be presynaptic in origin as evidenced by the lack of effect of toxin on responses to exogenous acetylcholine in the non-stimulated BCp.

Gln49-PLA_2能够剂量依赖性地抑制颈二腹肌神经信号正常传导并影响肌肉收缩,在100μg/ml剂量下,肌肉收缩幅度在150分钟后达到50%的抑制率;同时,随着Gln49-PLA_2浓度的降低,肌肉收缩幅度的抑制效率也逐渐降低;实验证明Gln49-PLA_2对突触后膜乙酰胆碱受体敏感性无影响,对神经肌肉信号传导收缩的抑制作用位点在突触前膜。

Results: BIO-1211 aerosol at 1, 3 and 10 mg/ml significantly inhibited the changes in lung resistance and lung dynamic compliance after antigen challenge in the sensitized rats in a dose-dependent manner, BIO-1211 at 25, 50, 100 and 200μg/ml inhibited the fibronectin-induced neutrophil adhesion by 23.5%, 24.6%, 61.4% and 58.1%, respectively, and serum-induced adhesion by 29.9%, 35.9%, 35.3% and 15.4%, respectively. Inhalation of 10mg/ml BIO-1211 did not show any protection against histamine and actetylcholin-induced bronchoconstriction.

结果:给大鼠气雾吸入BIO-1211 (1、3、10mg/ml)呈剂量依赖抑制致敏大鼠抗原攻击引起RL的升高和Cdyn降低;BIO-1211(25、50、100、200μg/ml)明显抑制纤维黏连蛋白和血清诱导的中性粒细胞黏附反应,其抑制率分别为23.5%、24.6%、61.4%、58.1%和29.9%、35.9%、35.3%、15.4%;但BIO-1211 (10 mg/ml)给豚鼠气雾吸入不能抑制乙酰胆碱和组胺诱导的药物性哮喘。

The preliminary bioassary showed that at the concentration of 100μg/mL, the inhibitory rate of compounds Ⅴc to Fusarium graminearum was 81.3%;Ⅳe to Fusarium oxysporium was 83.4%;Ⅴe and Ⅴf to Cercospora sorghi were 79.7% and 72.4% respectively;Ⅳd to Fusarium graminearum and Fusarium oxysporium were 68.9% and 65.7% respectively.

初步生物测试结果表明:在质量浓度为100μgmL下,化合物Ⅴc对小麦赤霉病菌的抑制率达到了81.3%;化合物Ⅳe对马铃薯干腐病菌的抑制率达到了83.4%;化合物Ⅴe,Ⅴf对玉米弯孢病菌的抑制率也分别达到了79.7%和72.4%;化合物Ⅳd对小麦赤霉病菌和马铃薯干腐病菌的抑制率分别达到了68.9%和65.7%。

The main findings of the study are as follows:(1) Both N2 and P3 are the characteristic ERPs under nogo inhibition,but P3 amplitude is a good indicator of inhibition processing depth;(2) Through ERP waveforms separation from encoding of stimuli sequence,we verify that the capacity of visual working memory is about 4 units;(3) In the go/nogo sequence response, encoding is mainly the function of right hemisphere, but inhibition retrieval is concentrated on left frontal area and anterior cingulated cortex.

本研究获得了以下结论:(1)在nogo抑制条件下,N2和P3波是特征的抑制波,但P3的波幅是反应抑制加工深度的良好指标;(2)对刺激的顺序编码加工的ERP验证了工作记忆的处理单元的容量为4左右;(3)在Go/nogo序列反应中,编码加工体现为右脑优势,而抑制提取的加工负波集中在左脑前额和前角回区。

The study showed the recombinant wt/mCREG protein depressed the VSMC proliferation depending on dose and the optimal concentration was 400nM;2biologic function of CREG protein and the membrance receptor mechanism:①effect on VSMC migration: the wound healing experiment showed the OB2 cells migration was slower significantly after added wt/mCREG(400Nm) in supernatant. The HITASY cells migration were very slowly and no remarkable change. The gelatinase digestion and Western blot analysis showed the matrix metalloproteinase was decreased and TIMPs was increased;②effect on differentiation: after added wt/mCREG(400nM), the expression of myocardin, SMα-actin, MHC and caldesmin were increased and that of LM-1 and FN were decreased in OB2 cells. These effects were more significant when adding wtCREG.;③effect on VSMC proliferation: Cell cycle assay and BrDU stain showed: after added the wtCREG and mCREG protein, the ratio of cell in G0/G1 phase increased to 0.5773 and 0.5572 from 0.5308 respectively in OB2 group, which increased to 0.7369 and 0.7034 respectively from 0.6297 in HITASY group;3Role of M6P/IGF2R in CREG biologic function:①ELISA and co-immunoprecipitation showed the wt/mCREG binding to M6P/IGF2R directly.②antibody blocking test: when the anti-IGF2R was added to medium at the same time with wt/mCREG at different concentration(2μg/mL、4μg/mL、8μg/mL),the effects of CREG protein which depressing proliferation, migration, secretion and promoting differentiation were blocked, which had the positive correlation to the concentration of added anti body. The studies showed two combinant CREG promoted VSMC switch to differentiation phaenotype, at the same time, depress VSMC proliferation, migration and secreting extracellular matrix.

上述实验结果证实:两种重组CREG蛋白对VSMC增殖均有剂量依赖性的抑制作用,并且相同浓度的糖基化的CREG蛋白对细胞增殖的抑制效应更为显著,最佳效应浓度为400nM;2两种重组CREG蛋白添加后对HITASY和OB2细胞生物学行为的影响:①CREG蛋白对VSMC迁移的影响:刮伤实验发现,加入最佳效应浓度的wtCREG和mCREG蛋白24h后,OB2组迁移能力下降,HITASY组无明显变化;细胞外基质金属蛋白酶-2,9(Matrix metallo-proteinase 2,9,MMP2 ,9)明胶酶电泳检测和Western blot检测结果证实,两种CREG蛋白均可以使OB2细胞合成细胞外基质MMP2,9减少,而组织金属蛋白酶抑制物(Tissue Inhibitors of Metalloproteinases,TIMPs)增加;②CREG蛋白对VSMC分化的影响:加入400nM的wtCREG和mCREG蛋白12h后,OB2细胞myocardin、SMα-actin、MHC、caldesmin表达增加,LM-1、FN表达减少;③流式细胞仪分析细胞周期和BrDU染色分析证实,加入400nM的wtCREG和mCREG蛋白后,OB2组G0/G1期细胞由0.5308分别增加至0.5773和0.5572,HITASY组G0/G1期细胞由0.6297分别增加至0.7369和0.7034;3M6P/IGF2R在重组CREG蛋白的生物学功能中的调控作用:①免疫共沉淀和免疫荧光双染色分析结果显示,CREG蛋白与M6P/IGF2R存在直接结合;②应用抗体阻断实验:将不同浓度的anti- M6P/IGF2R(2、4、8μg /mL)与两种CREG蛋白同时加入培养液中,CREG蛋白抑制VSMC增值、迁移和合成细胞外基质、促进分化的效应减弱,而且与加入anti- M6P/IGF2R浓度正相关。

The antibacterial activities of most of fractions were similar to the pomegranate peel extract.(7) Respiratory inhibition experiment showed that the respiration of thallus was inhibited mainly through the hexose monophosphate pathway, which leaded to be short of NADPH, and induced metabolic function dysfunction. Growth curve assay showed that the pomegranate peel extract inhibited cytodieresis in logarithmic growth phase. After exposure to the pomegranate peel extract, bacterial protein was analyzed by SDS-PAGE. The results showed that protein synthesis was inhibited, especially for large molecular weight protein.

7呼吸抑制试验结果表明,石榴果皮提取物主要是通过抑制HMP途径,导致还原力NADPH缺乏,引起代谢功能障碍,达到抑菌的目的;加药组和正常组金黄色葡萄球菌生长曲线的对比结果表明,石榴果皮提取物能够抑制金黄色葡萄球菌对数生长期的菌体分裂;SDS-聚丙烯酰胺凝胶电泳分析表明,石榴果皮提取物能够抑制菌细胞蛋白合成,尤其对菌体内大分子量蛋白作用最明显。

In T cells,TGF-beta1 can inhibit their proliferation,inhibit the differentiation of Th1 and Th2,inhibit the differentiation of CTL,induce the generation of regulatory T cellsTregin B cells,TGF-beta1 can inhibit their proliferation,induce IgA class switch,it also can induce the apoptosis of immature and resting B cells;in NK cells,TGF-beta1 can inhibit the secretion of IFN-γand the cytolytic function of NK cells.

对T细胞,TGF-beta1可以抑制其增殖,抑制Th1和Th2细胞的分化,抑制CTL的分化,诱导调节性T细胞的产生;对B细胞,TGF-beta1可以抑制B细胞的增殖,诱导IgA的类别转换,还可以诱导不成熟和静息B细胞的凋亡。

Lagenarium was higher 60%(for example, Glycyrrhiza yralensis, Lycium barberum and Desmodium styracifolium) as the concentration 0.01g/ml. Especially, the G. yralensis showed the strongest inhibitory effect on it with inhibitory rate of 99.6%. The inhibitory rate of 12 PE against mycelium growth and 15 PE against appressorium formation of C.

供试提取物在质量浓度为0.01 g/ml条件下,甘草、枸杞、金钱草等11种植物的80%乙醇提取物对黄瓜炭疽病菌分生孢子萌发的抑制率大于60%,其中甘草的抑制作用最强,达99.6%;12种提取物对黄瓜炭疽病菌菌丝生长的抑制率大于50%;15种提取物对黄瓜炭疽病菌附着胞形成抑制率达50%。

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The labia have now been sutured together almost completely.The drains and the Foley catheter come out at the top.

此刻阴唇已经几乎完全的缝在一起了,排除多余淤血体液的管子和Foley导管从顶端冒出来。

To get the business done, I suggest we split the difference in price.

为了做成这笔生意,我建议我们在价格上大家各让一半。

After an hour and no pup, look for continued contractions and arching of the back with no pup as a sign of trouble.

一个小时后,并没有任何的PUP ,寻找继续收缩和拱的背面没有任何的PUP作为一个注册的麻烦。