用

Took back my house yesterday, and refundment, everything was finished, the next step is upholster, the flooring of the living room and bedroom is food with cherry, it's good, and I checked online, we just polish, face0lifting, lacquering, that's enough, and it likes new floor, save much money for it, and I need to spend much money on kitchen and washroom, the current is so dirty which during more than 10 years, I will buy new cabinet and new shower room, it will be finished almost in 1 and half month, in the same time, all the fitment should be ready, everything is in control and on the schedule!

昨天把房子收回来了，钱也退了，接下来的就是装修了，又到装修的时候了，看了一下，客厅和卧室的地板用的是樱桃木的实木地板，这个我在网上查了一下，完全可以打磨，翻新，上漆等，和新的一样，光实木地板就省下不少钱，其他的大钱要用在厨房和厕所，统统敲掉，用了10来年了，都是油污和污垢，淋浴房做新的，买新橱柜，不过我的小厨房和小厕所也用不了太多的钱，装修用个1个半月差不多可以结束，同时家具和电器要到位，剩下的时间就是开始散味道了，一切事情按照计划进行，10月那么多人来上海就可以住得下了。

After two-cluster testing,branch length testing and relative rate testing ,two sequences(Melantitta nigra and Aythya americana) of 15 Cty b sequences were deleted because thek evolutionary rate were heterogeneous. The divergence between Anseriformes and Galliformes was chosen as an anchor point. Nucleotide sequence and amino acid sequence of 13 sequences data set was used to calibrated Anatidae molecular clocks. The results indicated that amino acid sequence is better than nucleotide sequence when divergence time is calculated with two kinds of sequences. The reliable divergence time of Anatidae were estimated with """"1+2TiTv"""" model.

经过二簇检验、分枝长度检验和相对速率检验，删除2条速率不恒定的序列后，用剩余的13条序列，以鸡形目与雁形目的分歧时间(89.8MYA)为锚定点，用顺序标定法标定鸭科鸟类分子钟的分析结果表明，用&1+2TiTv&分析得到的鸭科鸟类的分歧时间比较可靠；氨基酸序列用泊松校正模型和Γ模型估计的分类单元间的分歧时间则比较一致，表明氨基酸序列更适合用于鸭科鸟类分子钟的研究；用同一基因标定分子钟时，较长序列得到的结果标准误比较小；与核苷酸序列相比，氨基酸序列更适于用作分子钟的标定。

This is the problem of the low intermediary type of build or figure of the extremely dull standard that a reporter asks generally, I am early inside the answer themselves head that knows they hope to want had early, what group ah, parents, it is gibberish, the group is not to use benedictory, the group is to use what pay and support, parents also is not to use benedictory, parents uses love, acknowledgment has a fart to use, what the group needs is us pay and support, what parents needs is our love, not be what acknowledgment.

这是一个记者普遍问的极其无聊的标准的庸俗的&媒体型&的问题，我早知道他们希望要的答案他们自己脑袋里面早就有了，什么团队呀、父母呀，都是废话，团队不是用来感谢的，团队是用来付出和支持的，父母也不是用来感谢的，父母是用来爱的，感谢有个屁用呀，团队需要的是我们的付出和支持，父母需要的是我们的爱，不是什么感谢。

An allocation optical fiber sensor comprises signal treatment circuit, sensor optical cable, supernarrow wide laser, two detectors, three offsets, four couplers, four wave-length division multiplex, superradiation LED, polarizer and phase adapter, Sagna conoscope and Mach conoscope shares No1 sensitive optical fiber LB with the third wavelength division multiplex through the first wavelength division multiplex, the second wavelength division multiplex shared with the fourth wavelength division multiplex the second sensitive optical fiber to form the hybrid conoscope.

本发明公开了一种分布式光纤振动传感器，由信号处理电路、传感光缆、超窄线宽激光器、两个探测器、三个消偏器、四个耦合器、四个波分复用器、超辐射发光二极管、起偏器和相位调制器组成；在本发明中构成的萨格奈克干涉仪和马赫-泽德干涉仪，通过第一波分复用器与第三波分复用器共用第一敏感光纤L B ，第二波分复用器与第四波分复用器共用第二敏感光纤L C 形成了混合干涉仪。

Cells were immediately fixed and processed for immuno?uorescence and tested for translocation ofβ-catenin.β-catenin and target genes of Wnt/β-catenin signaling COX-2, cyclinD1, c-fos, c-Jun mRNA expression were examined by RT-PCR. Saos-2 osteoblastic cells were co-transfected with a wild type or mutant Tcf luciferase reporter gene and Renilla luciferase plasmid. Posttransfection cells were subjected to 1 hour of 12% elongation.

作用1h、2h和4h后，用免疫荧光的方法检测β-catenin的表达和在细胞上定位的变化；用半定量RT-PCR检测β-catenin mRNA表达的变化；用质粒转染的方法检测荧光素酶报告基因Tcf转录的情况；用半定量RT-PCR检测Wnt/β-catenin通路下游基因COX-2，cyclinD1， c-fos， c-Jun mRNA表达的情况；用免疫共沉淀的方法检测β-catenin和E-Cadherin的结合情况。

Based on the study, the paper discloses the nature and the characteristics of pragmatic meaning from the perspective of the definition of pragmatic meaning.( The pragmatic meaning of word refers to the temperative meaning of word which is given in application according to the conceptual meaning of a word, the purpose of users and the context it occurs.) Firstly, the pragmatic meaning is based on its static meaning and the premise of its existing is context in which it appears.

由此，从语用词义(词的语用意义是指在语言运用过程中，人们在词语概念意义的基础上，在一定的语用目的支配下，以语境作为参照而赋予词语的临时意义)的界定入手来揭示语用词义在具体交际中的性质及实际特征：一、语用词义存在的基础是词的静态意义，存在的条件是具体的语境。

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。

A brief pause to inquire of these "antis" if all Christians agree as to the meaning of their Teacher's message; all musicians as to the "correct" interpretation of a Brahms symphony; all doctors as to the "right" treatment of infantile paralysis; all parents as to the "perfect" way of raising children.

用一个简短的暂停问一下所有的反星象学者，是不是所有的基督信徒都是用同样的方式理解主的信息，是不是所有音乐家都用同样的&正确的&方法去诠释Brahms交响曲，是不是所有的医生都用同样的&正确的&方法治疗小儿麻痹症，是不是所有的父母都用同样的&完美&的方式去养育孩子。

The invention uses the fermentation liquor of one Escherichia coli generating polysialic acid as material, filters and removes thallus, adds some solvents as sodium chloride and alcohol into supernatant to deposit polysialic acid, dissolves the polysialic acid deposits via deionized water, filters to remove modified protein, uses alkali prolease to treat filtrate, complexes deposits via cetyl pyridinium chloride or cetyltrimethylammonium bromide, and dissociates, uses over alcohol to deposit polysialic acid, dissolves the deposits via ultra-pure water, grades and purifies via chromatography column, collects and dialyzes eluent, cools and dries to obtain polysialic acid.

本发明以一株产聚唾液酸大肠杆菌的发酵液为原料，经过过滤去除菌体，上清液加入适量氯化钠和乙醇等溶剂沉淀出聚唾液酸，聚唾液酸沉淀用去离子水溶解，再用过滤去除变性蛋白质，滤液经过碱性蛋白酶处理、氯代十六烷基吡啶或十六烷基三甲基溴化铵络合沉淀并解离后，再用过量乙醇沉淀出聚唾液酸，聚唾液酸沉淀用超纯水溶解，用层析柱进行分级纯化，收集洗脱液经过透析，冷冻干燥得聚唾液酸产品。

Among the ways that motorists have made this clear: spraying the cameras with paint; knocking them over; covering them in festive wrapping paper and garbage bags; digging them up; shooting, hammering and firebombing them; festooning them with burning tires; and filling their casings with self-expanding insulation foam that, when activated, blows them apart.

驾车者的下列行为清楚地表明了这一点：给相机喷洒颜料；撞倒相机；用喜庆的包装纸和垃圾袋将它们盖住；把相机挖出来；用枪射击、用锤子砸和用燃烧弹炸；用燃烧的轮胎在上面结彩；将自身膨胀的绝缘泡沫塞进相机外壳，一旦泡末膨胀起来，能将相机撑开。

Plunder melds and run with this jewel!

掠夺melds和运行与此宝石！

My dream is to be a crazy growing tree and extend at the edge between the city and the forest.

此刻，也许正是在通往天国的路上，我体验着这白色的晕旋。