经用

In our experiment, the GADes gene from rat brain was cloned and expressed in E.coli, so as to make the recombinant GADes produced by gene technology for clinical application and further study on the relation between GAD and type 1 diabetes mellitus.Firstly, with the pUClS/GADes recombinant plasmid as template, the full-length encoding sequence of GADes was amplified by PCR, and cloned into pGEM-T vector. Secondly, the target gene was cut by EcoRI and inserted into restriction sites of pET42a vector for expression.

本研究首先用PCR方法，以pUC18/GAD_(65)为模板，扩增目的基因，克隆到pGEM-T载体中，构建pGEM-T/GAD_(65)重组质粒，然后用EcoRI酶切，切下的目的基因片段插入pET42a融合型表达载体中，经酶切鉴定和测序，证实插入方向、读码框架正确；重组表达质粒转化大肠杆菌BL21(DE_3)中诱导表达，表达产物经亲和层析纯化后获得了融合蛋白GST-GAD_(65)。

Methods The total RNA was extracted from the brain of human embryo. The segment of open read frame of DOC-1 was amplified by RT-PCR and was further cloned into the vector, pMD18-T, and the prokaryotic expression vector, pGEX-4T-1, by turns, to produce the new construct pGEX-4T-1-DOC-1. After restriction enzymes digestion analysis and sequenced, pGEX-4T-1-DOC-1 was transformed into E.coli BL21（DE3）. GST-p12 recombinant protein was expressed under IPTG induction and purified by affinity column.

从人胎脑组织中提取总RNA，经逆转录-聚合酶链式反应扩增DOC-1的CDS序列，再通过基因重组技术将该基因片段依次克隆到pMD18-T和pGEX-4T-1载体中，构建融合表达载体pGEX-4T-1-DOC-1，经酶切、测序鉴定后，用该重组质粒转化E.coliBL21，用IPTG诱导表达，Glutathione Sepharose 4B柱亲和层析纯化重组蛋白。

After introducing 56Nb steel, in order to meet performance requirement of 56Nb steel fish plate on the railroad rail, the production technique of 56Nb steel fish plate has been adopted under the factory equipment conditions: 450 kg steel ingot is prepared by melting in 8 t arc furnace and casting at 1 570-1 580℃ which is heated to 1 250-1 270 ℃, trimmed to 185 mm×195 mm bloom after rolling by 600 series 2-stand rolling mill; then rolled 15 times by 600 series 2-stand rolling mill when steel ing

介绍了56Nb钢后，根据铁道接头夹板的性能要求并结合厂方的设备状况制定了56Nb钢鱼尾板材料的生产工艺：用8 t电弧炉冶炼，于1 570～1 580℃下铸成450 kg钢锭；将钢锭加热到1 250～1 270℃，经600系列2机架轧机5道次轧制后修整成185 mm×195 mm方坯；再将钢坯加热到1 050～1 150℃，由600系列轧机（2机架）共15道次轧制，用350系列（3机架）精轧机经3道次精轧成60 kg/m的鱼尾板材料，终轧温度约900℃。

Experimental mice were exposed to Trypanosome evansi after injection by different doses of antigenic reservoir. The results showed that 20 μg dose produced 83% and 67% protection to 2 sample variations after 4 immunizations. However, 3 immunizing doses could not give good protection. No protective effects were detected when experimental animals immunized by single VSG exposed to isotype from different rabbits.

用不同剂量抗原库免疫小鼠后进行攻虫，结果显示：经四次免疫后，20μg 剂量对随机抽取的两个变异体100 条虫体的保护率分别达到 83%和 67%；改变免疫方法，经三次免疫后的保护效果不理想；用单一 VSG 抗原免疫小鼠，接种来自不同兔体的同型虫体，不能起到相应保护作用。

The technique comprises the steps as follows: using the waste filtered mud of the sugar-refinery as the material to generate gas which is dehydrated till the water content is less than and equal to 30 percent and through distribution (clay used for of 70.00 to 85.0 percent, dehydrated filtered mud of the sugar-refinery of 15.0 to 30.0 percent, intensifier carboxymethyl cellulose or propyloic fibrin is 0.1 percent of the total dosage of the below material), performaing, moulding with high temperature (800 to 1200 DEG C, 20 to 180min) and acquiring the multi-hole porcelain granule with 20 to 30 percent of poriness, 10 to 20 percent of water absorbing capacity and 1.5 to 7.5/MPa of cylinder pressing intensity.

一种糖厂滤泥生产多孔陶粒技术，是用糖厂废弃物滤泥为发气原料，经脱水至含水≤30％，经配料(陶瓷用黏土70.0～85.0％，糖厂脱水滤泥15.0～30.0％，增强剂羧甲基纤维素或羧乙基纤维素为上述原料总量的0.1％)、制坯、高温(800~1200℃，20～180min)成型，得到孔隙率20~30％、吸水率10~20％、筒压强度1.5~7.5/MPa的多孔陶粒。

New reactions found in this program include: photoinduced regioselective free radical addition to aromatic imines;photoinduced Dielse-Alder reactions of aromatic imines for synthesis of tetrahydroquinoline derivatives;Photoinduced radical cation mediated selective ring opening reaction of epoxides;Formation of novel macrocyclic crown ethers by photoinduced reaction of diazonaphthaquinones;photoinduced selective desilylation of silyl ethers.

用这种方法成功的发现了下列在有机合成中有意义的新反应：(1)光诱导产生的自由基正离子诱导的自由基对芳香亚胺的区域选择性加成反应；(2)用光诱导的芳香亚胺的Diels-Alder反应合成四氢喹啉衍生物；(3)经光诱导产生的自由基正离子基诱导的环氧化物选择性开环反应；（4）重氮萘酮在环醚中经光诱导反应形成新奇的大环冠醚；（5）硅醚的光诱导选择性脱硅基反应。

Methods insert the stomach tube nasal cavity, with the help of intubating ferceps, take the stomach tube out of mouth cavity from the posterior wall of dysphagia, insert trachea catheter (cid 7.0~7.5) to esophagus, under the help of trachea catheter, insert the stomach tube to esophagus then take the trachea catheter out of the esophagus and vertical, cutting off, in the course of into esophagus.

经鼻腔插入胃管，用插管钳将胃管从咽后壁前取出口腔外，用id 7.0～7.5的气管导管插入食管，在气管导管诱导下将胃管经气管导管插入食管，然后将胃管向食管内送进，同时将气管导管退出，将胃管置入相当合适的深度后，固定胃管。

Methods: 2ethyl aniline was used as starting material of the synthesis and was processed by acetic anhydride acylation, strong nitric acid nitraction at low temperature, and desacyl synthesis to produce 2ethyl5nitro aniline. The product was then synthesized to 3methyl6nitro1Hindazole by using ringclosing reaction in the presence of NaNO2, and then ethylated to obtain 2,3diethyl6 nitro2Hindazole. The latter compound was then reduced by SnCl2/Hcl to get target compound 2,3diethyl6nitro2Hindazole.

以2乙基苯胺为原料经醋酐酰化、浓硝酸低温硝化及去酰基保护合成2乙基5硝基苯胺，后用亚硝酸钠关环得3甲基6硝基1H吲唑，经硫酸二甲酯甲基化得2，3二甲基6硝基2H吲唑，最后用氯化亚锡将其还原得到目标产物，通过1H NMR确定结构与目标产物一致。

Methods: 2-ethyl aniline was used as starting material of the synthesis and was processed by acetic anhydride acylation, strong nitric acid nitraction at low temperature, and desacyl synthesis to produce 2-ethyl-5-nitro aniline. The product was then synthesized to 3-methyl-6-nitro-lH-indazole by using ring-closing reaction in the presence of NaNO2, and then ethylated to obtain 2, 3-diethyl-6- nitro-2H-indazole. The latter compound was then reduced by SnC12/Hcl to get target compound 2, 3-diethyl-6-nitro-2H-indazole.

以2-乙基苯胺为原料经醋酐酰化、浓硝酸低温硝化及去酰基保护合成2-乙基－5-硝基苯胺，后用亚硝酸钠关环得3-甲基－6-硝基－1H-吲唑，经硫酸二甲酯甲基化得2，3-二甲基－6-硝基－2H-吲唑，最后用氯化亚锡将其还原得到目标产物，通过1HNMR确定结构与目标产物一致。

To prepare the phosphorylated PRAS40 (Ser183) antibody, We chosen 10-amino acid including Ser183 as antigen peptide through antigenicity and hydrophobicity analysis, hinged on keyhole limpet hemocyanin, and used the KLH-peptide to immunize rabbits. After antibody serum titer detection by enzyme linked immunosorbent assay, the antibody was purified with rProtein A sepharose fast flow and dephosphorylated antigen membrane. The antibody titrate reached 1:10 000 after purification and its special property was enhanced with absorption treatment of dephosphorylated antigen membrane. In addition, we used rabbit anti-PRAS40 antibody and the phosphorylated PRAS40 (Ser183) antibody to detect PRAS40 expression in several cell lines, including the normal cells HL7702, HEK293, tumor cells HepG2, A549 and S180. There were no quite difference among these cells; otherwise, we observed the decreased phosphorylation level of Ser183 after amino acid withdrawal treatment.

为了制备PRAS40(Ser183)磷酸化多克隆抗体，本实验通过蛋白疏水性抗原性分析设计多肽抗原，用其免疫家兔获得抗血清， ELISA检测其效价为1：10 000； Western blotting法检测发现，通过rProtein A Sepharose亲和层析纯化并经非磷酸化的抗原条吸附处理后的抗体可以明显提高磷酸化抗体的特异性；用PRAS40抗体及PRAS40(Ser183)磷酸化抗体对正常细胞HL7702、HEK293及肿瘤细胞HepG2、A549、S180的检测显示：磷酸化的Ser183在不同细胞中表达差异不显著，而在经细胞饥饿处理的HEK293细胞中却明显观察到了S183磷酸化水平随氨基酸含量降低而减弱的现象。

The split between the two groups can hardly be papered over.

这两个团体间的分歧难以掩饰。

This approach not only encourages a greater number of responses, but minimizes the likelihood of stale groupthink.

这种做法不仅鼓励了更多的反应，而且减少跟风的可能性。