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Design one primer pairs for EphB2 receptor and one primer pairs for GAPD as internal control. The RT-PCR was used to analyze the expression of EphB2 receptor tyrosine kinase in gastric cancer, colon cancer, lung cancer, liver cancer, kidney cancer, spleen cancer and abdominal wall cancer. The results were analyzed using Bio-RAD UV transilluminator with Quantity One software system.

采用RT-PCR方法，分别检测胃癌、结肠癌、肺癌、肝癌、肾癌、脾癌、腹壁癌7种肿瘤组织及相应邻近正常组织中EphB2受体的表达，应用Bio-RAD凝胶成像系统及在Quantity One软件辅助下分析测定表达结果。

We use 0.5% CMC to preserve the activity of Metarhizium anisopliae at room temperature. Then we carry on the RT-PCR semi-quantitative examination of the trehalase. The fresh Metarhizium anisopliae not preserved do control.

对用0.5%的羧甲基纤维素钠室温保存五个周的绿僵菌，进行海藻糖的半定量RT-PCR检测，用没有经过处理的固体培养基上培养14天的绿僵菌做对照。

We use 0.5% CMC to preserve the activity of Metarhizium anisopliae at room temperature. Then we carry on the RT-PCR semi-quantitative examination of the trehalase. The fresh Metarhizium anisopliae not preserved do control.

对用0.5%的羧甲基纤维素钠室温保存五个周的绿僵菌，进行海藻糖酶的半定量RT-PCR检测，用没有经过处理的固体培养基上培养14天的绿僵菌做对照。

In our research, the neutral trehalase gene in M. anisopliae was successfully cloned by PCR, RT-PCR, RACE and panhandle PCR, and submitted to GenBank, accession number are AY557613 and AY557612.

本研究利用PCR、RT-PCR、RACE、panhandle PCR 等方法成功地克隆了金龟子绿僵菌CQMa102 中性海藻糖酶基因，并登录NCBI 的GenBank，登录号为：AY557613， AY557612。

Six rats of each group were randomly sacrificed on day 7,14,28 and 42 respectively.The histological changes of lung tissue were studied by HE and Masson's trichrome staining.Hydroxyproline level in lung tissue was measured by digestion method.Protein and mRNA expression of transforming growth factor-β1（TGF-β1） were assayed by immunohistochemistry and RT-PCR respectively.

各组分别于造模后第7、14、28及42 d随机处死6只，取肺组织行HE、Masson染色评价肺组织病理变化，消化法测定肺组织羟脯氨酸含量，分别采用RT-PCR和免疫组化法测定肺组织转化生长因子β1（TGF-β1）的mRNA和蛋白表达。

Measure normal early pregnancy choria tissue and JAR cell by RT-PCR, LH/hCG receptor mRNA expression on both pregnancy choria tissue and JAR cell which is consistent with that be measured in ovarian granulosa cells, indicate that JAR cell and normal choria trophocyte presence LH/hCG receptor, hCG is one of the autocrine hormone.

二、用RT-PCR法可测到正常妊娠早孕绒毛组织、JAR细胞的LH／hCG受体mRNA表达，与正常卵巢颗粒细胞所表达的LH／hCG受体mRNA一致，提示JAR细胞与正常绒毛滋养细胞存在LH／hCG受体，hCG为其自分泌激素之一。

The model is as follows: WhereE = equity Rt = net annual cash flow in period t It = interest on mortgage in period t At = amortization of mortgage in period t Tt = income tax in period t Pn = selling price at end of period t = n GT = capital gains tax payable upon sale of property in year t = n UM = unamortized mortgage in year t = n n = investment horizon, in years from the present r = internal rate of returnThis equation can be solved by an iterative procedure of successive approxi mations to "r".

模范如下是：地方E=公平Rt=网年报现金流转在朝派时期t它=权益右手击球员的左后方场地抵押在朝派时期t阿特=amortization 的抵押在朝派时期t Tt=所得税在朝派时期t Pn=卖出价格阿特时期末t=n GT=首府gains税收可以支付在上面销售的财产在朝派年t=n嗯=分期偿还抵押在朝派年t=n n=投资视界在几年中从指向r=内部器官回报率这个等式罐被解决在附近一重复的过程的连续approximations向&r&。

It was found that after differentiation the cells expressed the endocrine markers insulin 1, glucagons, somatostatin by RT-PCR, however, these endocrine markers cannot be found in uninduced cells.

通过免疫细胞化学染色可以检测到在ICCs中存在有胰岛素、胰高血糖素、生长抑素和pdx-1阳性的细胞，通过RT-PCR可以在mRNA水平检测到胰岛素、胰高血糖素、生长抑素、pdx-1、ABCG2、葡萄糖转运蛋白等的表达。

Furthermore, preliminary work also performed to examine whether PI3K/AKT signal transduction pathway was activated in the process of refractory leukemia development. Materials and methods An immortalized human bone marrow stromal cell line, HS-5, was introduced to establish a bi-phase culture system for the cultivation of B-lineage precursor leukemia cells. ELISA and RT-PCR were used to investigate the expression of VEGF and its receptors in the leukemia cell lines and primary childhood leukemia cells in different treated groups. Flow cytometory method and immunofluorescent staining were employed to examine the apoptosis signals both in the VP16 treated and untreated leukemia cells. Western blot was utilized to explore the PI3K/AKT activated status in the drug induced or uninduced leukemia cells and lymphocytes from healthy donors.

材料和方法使用来源于人类骨髓基质细胞的细胞株HS-5作为滋养层细胞进行急性淋巴细胞性白血病细胞的体外培养，通过细胞生物学和免疫学方法评估培养体系并鉴定出难治性白血病细胞克隆；以ELISA和RT-PCR方法检测急性白血病细胞株和患儿白血病细胞VEGF及其受体的表达，了解不同治疗阶段VEGF及其受体的表达状况，并结合临床指标进行分析，明确VEGF及其受体在白血病发生过程中的作用；流式细胞仪和免疫荧光染色法对正常健康儿童、初发白血病患儿、复发白血病患儿及缓解后患儿进行凋亡因子检测和分析，初步阐明难治性白血病抗凋亡形成的原因；蛋白印记分析检测PI3K/AKT信号传导通路在健康儿童、初发白血病和复发白血病患儿的表达，初步了解难治性白血病形成的分子生物学机制。

Chicken IL-15(ChIL-15), Which was discovered in 2001, plays a main role in activating NK cells and CD8+ memory T cells, and may therefore have potential as a vaccine adjuvant.According to published ChIL-15 gene sequence, a pair of primers were designed and synthesized to clone ChIL-15 cDNA from ConA-activated chicken splenocytes by RT-PCR. Recombinant plasmid pUC19ChIL-15 carrying the ChIL-15 gene was constructed. The gene encoding mature ChIL-15 (mChIL-15) was amplified from pUC19ChIL-15 by PCR and cloned into pMD 18-T vector. After digested with Kpn I /Pst I , the mChIL-15 gene was subcloned into prokaryotic expression vector pPRoEX橦Ta and transformed into E.coli DH5a .The transformant was induced by IPTG, and the recombinant ChIL-15(rChIL-15) was expressed as a fusion protein with a histidine hexamer tag at the N-terminal end of the protein. Following expression, the protein was purified by ProBond?Nickel-Chelating Resin. The uninduced and induced protein lysates and the purified protein were separated by SDS-PAGE and transferred onto nitrocellulose membrane to identify rChIL-15 by Western blot. The rChIL-15 was used to immune the guinea pig to obtain rChIL-15 polyclonal antibody.

参考已发表的ChIL-15基因序列，设计合成引物，应用RT-PCR技术从刀豆球蛋白A活化的白来航鸡脾淋巴细胞中克隆ChIL-15 cDNA，将其与SmalⅠ酶切处理的pUC19载体连接，构建重组质粒pUC19ChIL-15；用PCR技术从pUC19ChIL-15中扩增出去信号肽的成熟ChIL-15(mChIL-15)基因，与pMD 18-T载体相连构建重组质粒pMDChIL-15；然后用KpnⅠ／PstⅠ双酶切下mChIL-15基因片段，定向亚克隆到经同样酶切处理的带有6个组氨酸标签的表达载体pP_RoEX~HTa中，构建重组质粒pP_RoChIL-15，对其测序确认读框正确后，转入大肠杆菌DH5α中诱导表达并进行纯化，用重组ChIL-15(rChIL-15)免疫豚鼠，制备多克隆抗体；再用HindⅢ／PstⅠ从质粒pP_RoChIL-15上切下mChIL-15基因，定向亚克隆到经同样酶切处理的杆状病毒转移载体pMelBacB中，经酶切、PCR和序列测定鉴定后，与杆状病毒线形化DNABac-N-Blue~(TM DNA共转染Sf9昆虫细胞，经三轮蚀斑纯化，构建重组杆状病毒rBac-ChIL-15，用该重组病毒感染处于对数生长期的Sf9细胞，按不同的时间收取样品，离心后对其上清和沉淀进行SDS-PAGE和Western blot分析。

Breath, muscle contraction of the buttocks; arch body, as far as possible to hold his head, right leg straight towards the ceiling (peg-leg knee in order to avoid muscle tension).

呼气，收缩臀部肌肉；拱起身体，尽量抬起头来，右腿伸直朝向天花板（膝微屈，以避免肌肉紧张）。

The cost of moving grain food products was unchanged from May, but year over year are up 8%.

粮食产品的运输费用与5月份相比没有变化，但却比去年同期高8％。