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We aim to screen and identify the key potential trans-factors during hemoglobin switch. We firstly analyzed differential expression of mRNAs in erythroid induction cultures of CD34+ cells derived from normal umbilical cord blood, adult bone marrow, and bone marrow of a heterocellular hereditary persistence of fetal hemoglobin patient. We identified ZF (HCF-binding transcription factor, Zhangfei) and SH3GLB1(SH3-domain GRB2-like endophilin B1) that had differential expression in the above three cultures. Furthermore, we confirmed the different expression of the above two genes by quantitative real-time PCR.

为鉴别影响珠蛋白基因表达和开关的新的重要调节因子，我们首先分析了人脐带血、正常成人骨髓和异细胞型胎儿血红蛋白持续存在综合症患者骨髓细胞红系诱导培养物中基因表达的差异，发现ZF(HCF-binding transcription factor，Zhangfei)和SH3GLB1（SH3-domain GRB2-like endophilin B1）在这三种不同来源的红细胞内具有显著的表达差异，并通过实时定量PCR方法验证了差异的真实性。

After the NK gene from recombinant plasmid pUC19-NK was cloned on expression vector pBV220, it was transformed into host cell E. coli HB101. Recombinant plasmid pBV220-NK was extracted for identifying by analysis of restriction enzymes, PCR and sequencing. Studies on recombinant strain's growth and expression showed that heterogene have no distinct effect on host cell. The recombinant plasmid have excellent segregational stability but low structural stability. The results of kinetics of pBV220-NK expression in E.

将重组质粒pUC19-NK上的纳豆激酶基因成功克隆到表达载体pBV220上，并将之转入大肠杆菌HB101中，得到转纳豆激酶基因工程菌，提取重组质粒经单酶切、双酶切及PCR分析验证后，对重组菌的生长及表达进行一系列研究，结果表明：外源纳豆激酶基因的导入对宿主的生长没有明显的影响：重组质粒的稳定性实验表明：该质粒具有良好的分离稳定性，而结构稳定性较差。

E studied expression levels of WAVE1 in six leukemia cell lines (the human Jurkat T leukemia cells, U937 histiocytic lymphoma cells, Burkitts lymphoma cell Raji, acute promyelocytic leukemia cell HL-60, chronic myelogenous leukemia cell K562 and K562/A02) by Western blotting analysis. Levels of WAVE1 expression were high in all six human leukemia cancer cell lines. In contrast, the constitutive expression levels of WAVE1 were noticeably lower in normal human peripheral blood mononuclear cells, or non-blood cancer cell-lines, including human lung A549 cancer cells, Hela cervical cancer cells, MG-63 osteosarcoma cells, CNE2 nasopharyngeal carcinoma cells, human umbilical vein endothelial cell, QSG7701 hepatocarcinoma cells, and WI-38 lung fibroblastoma cells.

AVE1在人类血液肿瘤细胞系(人T细胞白血病细胞系Jurkat、人组织细胞淋巴瘤细胞系U937、人Burkitts淋巴瘤细胞系Raji、人原髓细胞白血病细胞系HL-60、人慢性髓原白血病细胞系K562和K562／A02)中高表达，在非血液肿瘤细胞系(人肺腺癌细胞系A549、人宫颈癌细胞系Hela、人成骨肉瘤细胞系MG-63、人鼻咽癌细胞系CNE2、人脐静脉内皮细胞系HUVEC、人肺成纤维细胞系WI-38、人肝上皮细胞系QSG7701)以及正常人外周血单个核细胞中低表达或不表达。

PartⅡThe mechanism elucidation about effects ofα-(2,3)/(2,6)sialic acid on the Cx43gap-junction functions.(1) Westernblotting experiment showed that the decrease of sialic acid didn\'t changethe Cx43 expression and its phospholation level.(2) Westernblotting experiment showed sialidase didn,t change the ZO-1 expression,IP and confocal experiment showed sialidase improved the interaction of Cx43 andZO-1.(3) Westernblotting experiment showed sialidase didn\'t change N-cadherin expression,IP and confocal experiment showed sialidase promoted the complex formation ofCx43 and N-Cadherin.(4)Sialidase could increase the ERK1/2 phospholation level,and enchanedintercellular homotypic adhesion,Immunofluorometric assay showed sialidase couldpromote the N-cadherin cluster on the membrance.

第二部分：α—(2，3)／(2，6)唾液酸对肿瘤细胞CX43间隙连接功能影响的机制研究1、Westernblotting结果表明降低细胞膜表面唾液酸并不改变Cx43的表达及其磷酸化水平：但Cx43连接斑形成增多；2、Westernblotting结果表明唾液酸酶作用后细胞ZO—1表达没有改变，IP及免疫荧光共定位结果表明唾液酸酶作用后促进了Cx43与ZO-1的结合；3、Westernblotting结果表明唾液酸酶作用后细胞N-cadherin表达没有改变，IP及免疫荧光共定位结果表明唾液酸酶作用后促进了Cx43与N-Cadherin复合物的形成；4、唾液酸酶作用后细胞内ERK1／2磷酸化水平明显增加，细胞间同质粘附增加，以及免疫荧光表明唾液酸降低后可促进N-cadherin的膜成簇。

Results are as followed:1 Exposure of HELF cells to BP caused c-Jun activation,and increased the activity of MAPK,PI-3K,p53 and cyclin D1 pathway.2 BP-induced c-Jun activation was inhibited by dominant negative mutants of extracellular signal-regulated protein kinase or c-Jun NH_2-terminal kinase,but not by p38,impling that JNK and ERK pathways medicate c-Jun activation induced by BP.3 Overexpression of dominant-negative mutants PI-3K and Akt potently blocked phosphorylations of c-Jun and ERK,but not JNK in response to BP,suggesting that PI-3K/Akt pathway positively regulates BP-induced c-Jun activation through ERK.4 Inhibition of p53 by its chemical or molecular inhibitor markedly increased the phosphorylation levels of c-Jun,Akt and ERK upon BP stimulation,indicating that p53 negatively medicates BP-induced c-Jun activation through PI-3K/Akt/ERK pathway.5 The cell lines expressed TAM67 exhibits no significant affecting normal cell growth properties.6 TAM67 was able to significantly block G_1-S transition and subsequent cell proliferation,suggesting that c-Jun is essential for cell cycle alternations elicited by BP.7 Overexpression of TAM67 impaired BP-induced cyclin D1 activation,decreasing expression of E2F1 and pRb,indicating that c-Jun participates in the modulation of BP-induced activation of cyclin D1/pRb/E2F1 pathway.8 Stably expression of TAM67 led to the increases in the expression levels of p53 and p21,elevating phosphorylation level of p53,clearly indicating that c-Jun regulates p53/p21 pathway activation induced by BRCollectively,PI3K/Akt/ERK pathway mediated BP-induced c-Jun activation through p53-dependent mechanism.

结果显示：1BP刺激细胞可促进c-Jun活化，并伴随着MAPK、PI-3K、p53和cyclinD1通路各组成成分的活性增强。2利用MAPK通路的显性失活突变体分别阻断细胞外信号调节激酶和c-Jun氨基末端激酶活性，均可明显抑制BP诱导的c-Jun活化，但阻断p38活性对BP引起的c-Jun活化无明显影响，提示JNK和ERK通路参与调控BP诱导的c-Jun活化。3过表达PI-3K和Akt的显性失活突变体也可显著抑制BP诱导的c-Jun活化，并降低磷酸化ERK的表达水平，但对磷酸化JNK的表达水平无明显影响，说明PI-3K/Akt通路通过ERK正性调控了BP诱导的c-Jun活化。4p53的化学/分子抑制剂能使BP作用的细胞内c-Jun活性明显增加，并同时诱导Akt和ERK的磷酸化水平的升高，表明p53可通过PI-3K/Akt/ERK通路对BP诱导的c-Jun活化进行负性调控。5随后观察转染细胞的生长情况，发现TAM67对细胞正常生长和形态无明显影响。6稳定表达TAM67可有效抑制BP诱导的S期细胞数的增加，提示c-Jun在BP致细胞周期改变的过程中发挥了重要作用。7TAM67过表达能够抑制BP诱导的cyclin D1活化，降低磷酸化Rb以及E2F1蛋白表达水平，表明c-Jun参与调控BP诱导的cyclin D1/Rb/E2F1通路的活化。8过表达TAM67可使BP刺激的细胞中p53、p21总蛋白以及p53磷酸化的表达水平明显升高，可见c-Jun也参与调控BP诱导的p53/p21通路活化。

At present, the research about Heregulin-αand itsreceptors ErbB2 and ErbB3 was only the expression of them in mRNA and the impact ofHeregulin-αon the proliferation of mammary epithelial cells and milk proteins expression inculture. There were no results about the expression of Heregulin-αand its receptors in protein andthe impacts of Heregulin-αon the development, lactation and involution of mammary gland. It wasalso not clear about the signal transduction pathway of Heregulin-αin mammary gland.

目前对于乳腺中Heregulin-α及其受体ErbB2和ErbB3的研究仅限于其mRNA水平的表达以及Heregulin-α对培养的乳腺上皮细胞增殖和乳蛋白表达的影响，尚缺乏对乳腺中Heregulin-α及其受体在蛋白质水平上的表达变化规律以及Heregulin-α对乳腺发育、泌乳及退化的作用研究，正常乳腺中Heregulin-α的信号转导途径也不完全清楚。

Observation with transmission electron microscope showed that interalveolar septa were widened with increased amount of collagen in the MP infected rats while there were no obvious abnormalities in the other two groups.(2) Positive expression of bFGF mRNA were found in alveolar walls of MP infected rats. No expression of bFGF mRNA was found in control animals. In the rats infected with MP but treated with erythromycin positive expression of bFGF mRNA was found to be scarcely distributed in alveolar walls.

2感染组动物肺泡壁见明显的bFGF mRNA阳性表达颗粒，对照组基本未见bFGF mRNA阳性表达，感染加红霉素治疗组仅见散在分布的少许bFGF mRNA阳性表达；定量分析结果表明，感染组bFGF mRNA阳性表达的积分吸光度为41.32±10.44，与对照组(0.30±0.13)和感染加红霉素治疗组(6.03±2.41)比较，差异有显著性( P ＜0.01)。

The expression of MYP gene in gonad showed little difference between female and male. MYP gene expression was decreased rapidly in the gonad of S. intermedius at different stages, and slowly in hybrids. The comparative quantities of MYP expression in the gonads of S.

结果表明，MYP基因在海胆雌雄个体生殖腺中转录水平上的表达差异不明显；MYP基因在中间球海胆生殖腺发育的4个时期的表达呈快速下降的趋势，而在杂交海胆呈缓慢下降的趋势。

Despite evidence implicating prostaglandins as critical factor in male reproductive functions, little is known about expression and function of COX-2 in the male reproductive system. By using immunohistochemistry, RT-PCR and Western-blot, the content of this research included:(1) The expression and localization of COX-2 in mature rat and human testis were examined.(2) We investigated the changes of spermatogenesis and testosterone levels in testis after by using the COX-2 specific inhibitor-rofecoxib at 2, 4 and 8 weeks.(3) To explore the relationship between the intratesticular level of testosterone and expression of COX-2 by completely destroying the Leydig cells of mature male rats with injection of a single I.

本课题应用免疫组织化学、RT-PCR和Western-blot等方法检测以下内容：(1)在mRNA和蛋白水平检测正常成年男性及大鼠睾丸组织中COX-2的表达及定位情况；(2)实验组大鼠持续给予COX-2酶的特异性抑制剂Rofecoxib，分别于用药后2周、4周和8周检测大鼠的精子生成以及睾丸组织内睾酮浓度的变化；(3)另一组实验大鼠给予特异性Leydig细胞灭活剂EDS，在特定时间内抑制睾丸组织内合成的睾酮，于用药后2周和4周，采用免疫组织化学及RT-PCR方法观察大鼠睾丸组织内睾酮浓度变化对COX-2合成的影响。

The expression of PTEN protein was detected massively or lamellarly only innuclei. IOD represents the expression of it when the expression PTEN and MGMTprotein was investigated by confocal laser scanning microscope.

在激光共聚焦显微镜观察PTEN和MGMT蛋白在食管鳞癌组织中的表达时，用积分光密度值表示PTEN和MGMT蛋白表达的量。

In contrast to the ubiquitous rising-sun-with-rays military flag of the Japanese, Chinese banners and ensigns feature a range of designs.

与遍地都是的太阳军旗不同，中国人的旗帜和徽章设计得各式各样。

From their small corner of Feng's Guangzhou headquarters -- a jumble of pink leashes, squeezable rubber steaks, and plastic doggy Santas for Fido's stocking -- Soleil's designers come up with at least five new products a month.

从Feng 设在广州总部的产品展示柜台上可以看到，Soleil的设计师每月至少设计出5件新产品。