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In order to reduce the non-specific binding between oligonucleotides and bases mutation caused by the complicate secondary structure of DNA and excessive PCR amplification, a frequently phenomenon in one-step gene synthesis, we used a two-step method including assembly PCR and digestion-ligation step to synthesis Aspergillus niger lipase gene lipA.

本研究主要针对一步法长片段基因合成中复杂结构的非特异性配对和过多的PCR引入碱基错配等问题，采用二步法(组装PCR和酶切-酶连)合成了黑曲霉脂肪酶基因lipA。

a three steps sequence therapy was designed for patients suffered post dural puncture headache total 45 cases:step one,patient keep supine recumbence,and intravenous injection of ringer's solution 1000ml with intramuscular injection of tramadol 100mg was administrated. if pdph could not relieve within 24hr,the next step should be use. step two,drinking a cup of coffee three times a day for patients who were allowed to eating or intravenous injection ketamine 0.5mg/kg for patient in fasting diet.

选择椎管内麻醉后发生头痛的病例共45例，按设定的序贯疗法分别为a：去枕平卧1d，每天补液量增加1000ml平衡液，肌注曲马多100mg，无效时进入b和c；b：能进食者给予市售咖啡饮品，每天三次；c：不能进食者静脉注射氯胺酮0.5mg/kg，连用2d；以上治疗无效时进入d：行硬膜外填充自体血10ml。

The officers and men marched in the parade square standard pace, each step of 75 cm, 116 steps per minute, did Folks; stringent requirements, rigorous training to make phalanx of officers and men trained in a real skill. 56 Fang team, a symbol of unity and harmony of China's 56 nationalities, unite as one, making big strides along the road of socialism with Chinese characteristics; flag guard members from the People's Heroes Monument to the starting base of the flagpole to reach the road during the process of step goose-step 169 , meaning "from 1840 to the present 169 years of extraordinary history, the Chinese bull by the horns with confidence towards the future"......

阅兵式方阵的官兵们迈着标准的步伐，每一步75厘米、每分钟116步做到了分毫不差；严谨的要求、严格的训练使方阵的官兵们练就了一身的真本领。56个方队，寓意着我09年国56个民族团结和谐、万众一心，沿着中09年国特色社会主义道路阔步前进；09年国旗护卫队队员从人民英雄纪念碑出发到达09年国旗杆基座的行进过程中将迈出正步169步，寓意&从1840年至今169年的不平凡历程，中华民族不畏艰难，满怀信心，走向未来&

The method includes the following steps: step A, the terminal accesses, registers and loges on to IM core net through at least two different wireless IP access networks; step B, in the process for carrying a VoIP communication through at least one of two IP access networks, if the terminal meets the pre-set switching condition between the IP access network, a switching request in the IMS domain is initiated; step C, the speech calling continues application server receives the switching request, and switches the signaling between the terminal and the VCC application server to the other IP access network except the IP access network loading the VoIP communication at present in at least two IP access networks.

本发明提供一种IMS域内VoIP的切换方法、IMS系统、终端及VCC应用服务器，该方法包括：步骤A，终端通过至少两个不同的无线IP接入网接入并注册登录到IMS核心网；步骤B，在终端通过至少两个IP接入网中的一个进行VoIP通话的过程中，如果终端满足预设的IP接入网之间切换条件，则发起IMS域内的切换请求；步骤C，语音呼叫连续应用服务器接收切换请求，并将终端与VCC应用服务器之间的信令切换到至少两个IP接入网中、除当前承载VoIP通话的IP接入网之外的其中一个IP接入网上。

One Of Us They passed me by, all of those great romances You were, I felt, robbing me of my rightful chances My picture clear, everything seemed so easy And so I dealt you the blow One of us had to go Now it's different, I want you to know One of us is crying One of us is lying In her lonely bed Staring at the ceiling Wishing she was somewhere else instead One of us is lonely One of us is only Waiting for a call Sorry for herself, feeling stupid feeling small Wishing she had never left at all I saw myself as a concealed attraction I felt you kept me away from the heat and the action Just like a child, stubborn and misconceiving That's how I started the show One of us had to go Now I've changed and I want you to know One of us is crying One of us is lying In her lonely bed Staring at the ceiling Wishing she was somewhere else instead One of us is lonely One of us is only Waiting for a call Sorry for herself, feeling stupid feeling small Wishing she had never left at all Never left at all Staring at the ceiling Wishing she was somewhere else instead One of us is lonely One of us is only Waiting for a call

我们其中之一他们让我通过，所有那些重大的浪漫史我感觉，你是剥夺我的公正机会我的画很清除，一切看起来这样容易的所以我给予你打击我们其中一个必须走我想要你知道，现在已经不同了我们其中一个在哭泣我们其中一个在说谎在她孤独的床凝视天花板希望她倒是在别的地方我们其中一个是孤独的我们其中一个只是等侯一个电话对自己感到遗憾，觉得愚蠢觉得渺小希望她没有完全离去我把我自己视作一个被隐藏的吸引我觉得你离开热度和行动的我就像一个孩子一样，顽固和误解那就是如何我开始展览会我们其中一个必须去现在我改变了想法，我想要你知道我们其中一个在哭泣我们其中一个在说谎在她的孤独的床凝视天花板希望她倒是在别的地方我们其中一个是孤独的我们其中一个只是等侯一个电话对自己感到遗憾，觉得愚蠢觉得渺小希望她没有完全离去了没有完全离去凝视天花板希望她倒是在别的地方我们其中一个是孤独的我们其中一个只是等侯一个电话

Children have taken the first step to prepare a timely manner after the affirmative, the second step, the third step can be easily handled.

孩子有准备地迈出第一步后，及时肯定，第二步、第三步就好办了。

Meanwhile, the concentration of Pb,Zn and other heavy metals is rather high in the oxidation zone and decreasesdownward to levels that are unbelievably low for Pb-Zn mine tailings. Theseobservations suggest that heavy metals should be re-immobilized by secondaryminerals, rather than primary minerals, in the oxidation zone, and could be leachedout from suboxic environment due to low rate oxidative dissolution which preventsthe precipitation of secondary minerals in significant amount. Thus, cover scenarioswhich establish a suboxic environment beneath the cover layer might not always beeffective in preventing heavy metal release from mine tailings.Sequential extraction studies indicate that, in addition to heavy metal leaching,oxidation of the mine tailings increased the mobility of residual Zn, and Cu, while Pb remains immobile due to the formation of anglesite (PbSO4). Using the seven-step extraction technique developed by Dold (J. Geochem. Explor., 2003, 80: 55-68), galena dissolves in significant amount at step 2 most probably due to the strong complexing of Pb2+ by acetate, while other metal sulfides dissolve at step 6, which suggest that this extraction technique could not be used to evaluate the mobility of lead in natural environment

重金属含量均低于新鲜尾矿，但以硬化层中的含量最高(Pb、Zn含量分别为0.47%和0.93%)，往下大幅度降低，表明尾矿风化过程中形成的次生矿物对Pb、Zn等重金属元素有较强的固定作用，而次氧化带的淋溶作用造成了金属元素的大量流失；(4)与新鲜尾矿相比，硬化层中铅的可迁移性大大降低，硬化层之下尾矿样品中铅的可迁移性也不同程度降低，这是由于硫化物氧化产生的硫酸根与尾矿中的铅反应形成铅矾(PbSO_4)的缘故；(5)氧化作用使尾矿中的zn、Cu等重金属的可迁移性不同程度地增大，其中对锌的可迁移性影响最大，铜的可迁移性与铁比较相似，铅则容易与硫化物氧化形成的SO_4~(2-)反应形成铅矾(PbSO_4)；(6)为了减少尾矿污染和修复矿山环境，人们提出了各种不同的尾矿覆盖方案，本文的研究结果表明，这些方法不一定能够有效地阻滞污染元素的迁移。

Statistic analysis result shows that the time step is the most important influence factor of simulation precision, and there is relation between simulation error and the setup value of restitution coefficient which the greater the setup value is, the less the simulation error is. Although there is no effect to the total contact time in the course of collision, the value of the time step decides the total contact step, and there is a logarithmic linear correlativity between them.

统计分析结果表明：计算时步是影响模拟精度最重要的因素；模拟误差还同恢复系数的设定值有关，设定值越大，模拟误差就越小；计算时步的大小虽然对碰撞过程的总接触时间影响不大，但是却决定了碰撞过程的总接触时步数，二者之间存在对数线性相关关系

With the reform step of the commercial bank has expedited, new institutional and constant renew from time to time, conditionality system is step by step to perfect, the management of the fixed assets is request more and more.

随着商业银行改革步伐的加快，新制度的不变更新，相应制约机制的不断完善，对固定资产的管理提出了新的要求。

Carried on my backpack, grasped my crabstick, move up step by step carefully, every step, the race made me toppleing, the river seemed to show the last stateliness.

打好背包，抓紧手杖一步一步小心翼翼向前挪动着脚步，每踏出一步，汹涌的急流就冲得我摇摇欲坠，好象在向我显示着它最后的威严。

A historical region of southwest France between the Pyrenees and the Garonne River. The duchy of Aquitaine was joined with France after the marriage of Eleanor of Aquitaine to King Louis VII in1137, but its possession was disputed after her subsequent marriage to Henry II of England.

阿基坦法国西南部一个历史地区，位于比利牛斯山和加隆河之间，在1137年阿基坦的埃莉诺与国王路易斯七世结婚之后阿基坦公国加入法国，但当她再嫁英国亨利二世后，其归属权便受到争议

There are moments when I look at the smiling , happy faces of my expectant staff and wonder how they will cope when confronted by an outraged housemaster along with an unrepentant and recalcitrant bleached blond teenager in 18 years ' time .

有时候，我看着那些满怀期待的员工微笑，幸福的脸，想着当他们遇到一位愤怒的宿舍管理员和一位不知悔改，叛逆，而且将头发染成白色的18岁少年时，他们会如何处理。