双染的

Let G be a double-outer plane graph, V, E, F be the set of points, edge and face, a plane graph G is k-total colorable, if the elements of V E can be colored with K-colors, any two distinct adjacent or incident elements receive different colors.

设G是一个双外平面图，V，E，F分别为双外平面图G的点集，边集和面集。G的全色数X是使得V∪E中的任意两个相邻或相关联的元素间均染不同颜色的最少颜色数。

We describe the specific silencing of reporter gene lacZ in FHM cells (muscle cells of fathead minnow, a fish cell line) by either expressing small hairpin RNAs from plasmids or transfecting small interfering RNAs transcribed in itro.

在FHM细胞内通过转染质粒表达小发夹RNA或者直接转染小干扰RNA能特异沉默报告基因LacZ的表达，且siRNAs 抑制效果更加明显；这两种类型的小双链RNA在Hela细胞内同样能抑制报告基因的表达。

Nowadays in India, the true orthodox Buddhism has been completely extinguished with only a few superficial appearances left, and the substance has been replaced by the non-Buddhist tantrism of couple-practice. About Theravada Buddhism, although it can still keep the appearance of sound-hearer Buddhism and is not defiled by the non-Buddhist tantrism, it is rare to find the saints who can personally realize the sound-hearer liberation now. As for Mahayana Buddhism, there are only Buddhist rituals and the mind-consciousness dharma now in China, which has just recovered from the ravage of the Cultural Revolution. In both Japan and Korea, only the appearance of Buddhist rituals but not practice remains and there is no even the mind-consciousness practice like in China. Therefore the practitioners in China, Japan and Korea cannot touch the real content of the practice and realization to fulfill the Buddhahood way of the great-vehicle Buddhism.

现存的佛教，在印度已经完全灭亡，只余极少数人保留著佛教的表相，本质已是坦特罗双身法的非佛教了；南传的小乘佛教教域，虽然仍能维持小乘声闻佛教的表相而不被坦特罗双身法的非佛教所染污，然而能够实证声闻解脱道的圣者亦已难可觅得；而北传的大乘佛教教域中，中国才刚从文化大革命的破坏中恢复了仪式表相及意识境界的佛教，日本、韩国则只剩佛教仪轨的表相而无修行的表相，已谈不上如同中国地区意识境界的修行了；是故中、日、韩对於大乘佛教实践菩萨道的见道与修道真实内涵，都已经都无力触及了。

The CCND2 expression cell proliferation, cell cycle and cell apopotosis of the transfected LP-1 cells were studied by RT-PCR, trypanosome staining, flow cytometry and annexin V assay.

设计、合成CCND2短发夹RNA双链DNA模板，构建表达质粒pGenesil-2／CCND2，转染LP-1细胞，以RT—PCR检测对CCND2表达的抑制作用，以锥虫蓝染色计数和MTI检测细胞增殖，以Annexin V检测细胞凋亡，流式细胞术检测细胞周期。

Dual-Luciferase Reporter Assay was used to demonstrate the activation of this regulatable vector after cotransfection with pRL-TK in HEK293 cells.

在体外转染HEK293细胞后，运用双荧光素酶报告基因技术鉴定了该系统的调控能力。

Methods The total RNA was acquired from young rabbit particular cartilage, the BMP7 gene was inserted into pcDNA-3.1 to construct eukaryotic expression vector plasmid of pcDNA3.1-BMP7; the target gene-BMP7 was identified respectively by PCR analysis, restriction enounces analysis and nucleotide sequencing; the plasmid was transfected into adult rabbit knee articular chondrocytes, then the expression of BMP7 was detected by in situ hybridization, PCR and Western blotting.

从体外培养的幼兔膝关节软骨细胞中提取总RNA；按GenBank BMP7基因序列化学合成2条引物，采用RT-PCR方法得到BMP7基因；将BMP7基因片段插入到真核表达载体pcDNA 3.1中，构建pcDNA3.1-BMP7真核表达载体质粒；利用双酶切、PCR及核苷酸序列分析鉴定BMP7目的基因；将重组pcDNA3.1-BMP7真核表达载体质粒转染家兔关节软骨细胞，分别采用原位杂交、PCR、Western blotting法测定BMP-7表达。

Us, who the world with bloody stain imbrued

来拜访我们这用鲜血染红世界的一双

The PCR products were examined by agarose gel electrophoresis. The target gene fragments were purified by gel extraction kit and ligated to cloning vector pMD18-T. The recombinant vectors were transformed into host strain E. coli K802 by lithium chloride method, screened and identified with PCR and restrictive enzymatic digestion. Their sequences were confirmed by DNA sequencing.(2) sTWEAK1 gene was subcloned into expression vector pProEx HTb and transformed into E. coli BL21. sTWEAK2 gene was subcloned into expression vector pMAL-C2x and transformed into E. coli TB1. The recombinant vectors were screened and identified with PCR and restrictive enzymatic digestion. The recombinant fusion proteins were induced to express with IPTG, detected by coomassie brilliant blue-stained SDS-polyacrylamide gel electrophoresis , and confirmed by Western blot analysis.(3) The sTWEAK1 fusion protein was purified with Ni-NTA Spin Kit.(4) The biological activity was assayed on transformed and tumor cells by microplate photometer after crystal violet or sulfur rodamine B staining.(5) The contents of IL-8 in the supernatant of 1990 cell cultures were determined by ELISA.(6) The morphological changes of the sensitive cells were observed by light and transmission electron microscopies.(7) The cell cycle and apoptotic rate were assayed by flow cytometry in 1990 and M85 cells.(8) The effect of fusion proteins on induction of NF-κB in 1990 and LOVO cells was detected with Dual-Luciferase Reporter Assay system.(9) The TWEAK gene was subcloned into Adeno-X Viral DNA with pShuttle vector and transfected into HEK293 cells by lipofectamine method.

(1)本研究用RT-PCR方法，从人组织细胞总RNA中扩增可溶性TWEAK胞外区（sTWEAK1和sTWEAK2）的cDNA序列及全长编码序列，用琼脂糖凝胶电泳分析PCR产物，胶回收目的基因片段，连接到pMD18-T克隆载体中，转化大肠杆菌K802，PCR和酶切筛选阳性克隆，全自动DNA测序验证序列；(2)sTWEAK1和sTWEAK2分别亚克隆到pProEx HTb和pMAL-C2x表达载体中，分别转化大肠杆菌BL21和TB1，PCR筛选和酶切鉴定，阳性克隆用IPTG诱导表达，表达产物用SDS-PAGE分析和Western blot验证融合蛋白；(3)用NTA-Ni Spin试剂盒初步分离纯化sTWEAK1融合蛋白；(4)用体外培养的肿瘤细胞和正常对表达产物进行活性检测，贴壁细胞用结晶紫染色法，悬浮细胞用磺酰罗丹明B染色法，酶标仪检测OD值；(5)敏感细胞用ELISA法检测细胞培养上清中IL-8的含量；(6)用光镜和电镜观察敏感细胞死亡和细胞凋亡情况；(7)用流式细胞仪分析表达产物对敏感细胞凋亡率和细胞周期的影响；(8)用双荧光素酶报告基因检测法，测定表达产物对敏感细胞NF-κB的影响；(9)用pShuttle穿梭质粒将TWEAK重组到腺病毒载体上，用脂质体转染法转染HEK293细胞，PCR鉴定重组质粒。

From the investigation into the visible spectra and performances on textile of these azo dyes, as acid dye, it was found that these dyes had deep coloring effects on dyeing wool or silk, and five of which could dye wool into black one at 3% o. w. f., especially the diazo dye derived from 2, 8-diamino-10-methoxy-5, 10-dihydrophenophosphazine 10-oxide and acet-J-acid, it could not only dye wool into black one but each fastness of the obtained dyed sample was good.

对这些已知结构的染料进行浸染染色，结果发现，它们作为酸性染料，在羊毛及丝绸纤维上染色时，能够获得比一般偶氮染料不宜获得的深色效应，其中5支染料得到黑色染样，尤其由2，8-二氨基-10-甲氧基-5，10-二氢磷杂吖嗪与乙酰J-酸形成的双偶氮染料得到黑色染样，其各项牢度都比较理想。

For the two kinds of fibre, each uptakes complementary colours respectively, thus obtaining two colour effects and incresing added-value of the product.

探讨了高密细旦涤纶与棉交织物的染整加工工艺，采用溢流染色机分别对细旦涤纶和棉上染不同颜色，获取特殊的双色效果，提高了产品的附加价值。

For a big chunk of credit-card losses; the number of filings (and thus charge-off rates) would be rising again, whether

年美国个人破产法的一个改动使得破产登记急速下降，而后引起了信用卡大规模的亏损。

Eph. 4:23 And that you be renewed in the spirit of your mind

弗四23 而在你们心思的灵里得以更新