双切的

The main environment geological question is: The earth"s crust where the faultage and earthquake are active is unstable; collapse , landslide, mud-rock flow and soil erosion ; The underground karst subsides, seepage question; Basic rock expand, expend and compress soil body, the salt deposit corrode out of shape and groundwater"s corrodent harm; The soft soil question of basin; The soft rock body, cracked rock and the weathering rock are relatively poor stability; High and cold regional highways and railways" frozen soils freeze and melt calamity problem; The around rock of tunnel are unstable because of the coal seam gas, spring water, underground developed area; Project cutting slopes, constructed abandon soil and reservoir, lake"s ecology geological environmental protection issue; Thedangerous shoal of the channel % submerged reef question; The problem of rebuilding channel project and dock etc.; And the problem of unstable ground and groundwater corrosivity during building airport; At the same time, with the international big pathways" implementation and completions of constructions, adjusting the cities and counties" constructions , the crowd occupy changing , the cultivated land distribute changing and the adjustments of structure, will cause local environmental geological issues outstanding; According to multiple statistical analysis , value calculate and integrated appraise result, in the northwest and southwest of Yunnan, the traffic relatively low density, traffic engineering is relatively weak impact on environment, It is the area where a environme

主要的环境地质问题是：活动性断裂、地震带的地壳不稳定；崩塌、滑坡、泥石流及水土流失；地下岩溶塌陷、渗漏问题；基础岩体膨胀、胀缩土体、含盐层侵蚀变形和地下水的腐蚀危害；盆地软土问题；软弱岩体、碎裂、风化岩体稳定性较差；高寒地带公路、铁路建设的冻土冻融灾害问题；煤层瓦斯、涌水、地下采空区等隧道围岩不稳定问题；工程切坡、施工弃土及水库、湖泊生态地质环境保护问题；航道险滩、暗礁问题；渠化工程、码头等库岸再造问题；以及机场建设中的不稳定地基及地下水腐蚀性问题；同时，随着大通道建设的实施和完成，城镇建设的调整、人群居落的变化和耕地分布及结构的调整组合，都可能造成局部环境地质问题的突出等等。经多元统计分析数值计算、综合评价结果，滇西北、滇西南地区交通密度较低，交通工程对环境的影响程度较弱，是环境地质状况好的区域；有主要高原湖泊分布区的，包括昆明、个旧、文山的滇东南区域，环境地质状况较好；大姚、楚雄、篙明及会泽、昭通一镇雄的区域，即滇中北部中生界红层和滇东北岩溶区，环境地质状况中等；而包括保山、德宏、大理、临沧的滇西地区及景东一墨江以东、双柏一石屏一河口以西及东川一寻甸一曲靖地段的滇中一滇东地区，环境地质状况较差。云南国际大通道建设涉及全省区域，如何利用地质环境、实现可持续发展，就必须依赖于国际大通道建设与地质环境之间良性关系的建立。应本着对区域地质环境客观存在的科学认识原则、建设过程中环境效益优先的可持续发展原则、法制性原则、对大通道建设中环境地质的因地制宜及其可防治性原则。并且从组织管理、不同类型大通道、不同环境地质问题类型等方面，提出了对策措施。最后，提出了建立国际大通道建设与环境地质良性关系的宏观建议。

Renli fully satisfies the requirements of Implementation of Surgical Implants Production. It's equipped with a complete set of equipments for vacuum casting processing line, such as vertical processing center, horizontal digital control lathe, digital control straight-cut automatic lathe, vertical digital control milling machine, four-column hydraulic pressure machine, a complete double-die automatic cold header, electric cutting machine,laser labeling machine, intellectual pneumatic labeling machine, bar code labeling machine, etc., totally over 130 sets of automatic and/or general equipments. It also owns 10,000 class clean rooms and relevant manufacturing and monitoring equipments. The sterile packaging

公司已完全具备《外科植入物生产实施细则》的要求，拥有真空铸造生产线的整套设备；拥有数控立式加工中心、卧式数控车、数控纵切自动车、立式数控铣、四柱液压机、双摸整机自动冷墩机、电火花切割机、激光打标机、智能气动打标机、条形码标签机等各类高精自动设备和通用设备 130 余台套；具备 10 万级洁净厂房和相应配套的生产和质量监控设备，可在封闭洁净厂房里操作外科植入物产品的无菌包装；具有满足生产全过程和最终成品的金属材料化学成份分析、力学性能检测、硬度检测、金相检测、粗糙度检测、表面缺陷检测、形位曲面检测等的理化检测仪器和设备，能有效控制产品的质量。

Results We obtained a lysogenic bacteriophage MZTP01with clear plaque and 1 mm diameter. Fragment D with 2362bp (Genebank No. AY639599) was obtained after the phage DNA hydrolyzed by HindⅢ/EcoRⅠ. Among fragment D, the pep gene with molecular weight of 47 kDa and length of 1101bp was cloned and expressed. Recombinant M15 (pQE30pep) was built and overexpressed in Escherichia coli with a 47kDa clear band. At the same place a clear band was observed by Western blot. Judging from the time course expression, we could conclude that PEP protein produced at 1 hour after induction and then increased gradually. PEP protein was mainly in the form of inclusion body in the recombinant and slowed the growth speed of host. Homologous comparison of PEP protein from phage MZTP01 with other PEPs from BLAST were that phage MZTP01 PEP protein had 100% homologe with that of Escherichia coli K12, and most of others took the similarity in the range between 37%~84%.

诱导获得的溶原性噬菌体MZTP01斑点清晰，直径约1mm，成斑时间12h；从噬菌体基因组DNA双酶切(HindⅢ/EcoRⅠ)片段中回收长度为2362bp的D片段(Genbank登录号： AY639599)，又从D片段中克隆了长度为1101bp、编码367aa、分子量为47kDa的pep基因，表达载体M15(pQE30pep)在大肠杆菌(Escherichia coli， E.coli)中表达获得了47kDa的清晰表达带，在1h 时开始产生蛋白并有逐步上升的趋势； Western blot 也在47kDa处得到一条清晰的条带；可溶性分析表明PEP蛋白在重组菌株中是以不可溶的包含体形式存在的，该蛋白的产生明显地抑制了宿主的生长速度；噬菌体PEP氨基酸序列之间的同源性比较表明，噬菌体MZTP01 PEP蛋白与来自E.coli K12噬菌体的PEP蛋白的同源性程度最大。

To systematicallyinvestigate this landmark, we conducted a study in four phases:1 Two blindfolded examiners determined the distance betweenthe tuberosity's medial border and the clavicle's lateral endin 100 dried clavicles and then 2 performed subclavian veincannulation in 20 fresh human cadavers using the tuberosityand the suprasternal notch as landmarks. 3 Three-dimensionalreconstructions of the subclavian artery and vein and surroundingstructures were derived from computed tomography datasets of10 patients. The length of the path of a virtual subclavianvein cannulation with the deltoid tuberosity landmark was measuredbilaterally. 4 In a prospective, randomized trial, subclavianvein cannulation was performed in 60 patients with a standardapproach or with the deltoid tuberosity as landmark.

为系统地研究这个解剖标志，作者将本研究分为四个阶段：1 双盲检查100个干燥的锁骨测量其三角肌粗隆内侧缘至锁骨侧面末端的距离；2 在20具新鲜的尸体上以三角肌粗隆和胸骨上切迹为标志进行锁骨下静脉穿刺；3 利用 CT 所获取的数据资料对10个病人的锁骨下动脉和静脉以及周围结构进行三维重建，测量两侧以三角肌粗隆为标志的虚拟锁骨下静脉置管路径的长度；4 选择60例病人以标准路径或三角肌粗隆为标志进行锁骨下静脉穿刺的前瞻性随机试验研究。

To systematicallyinvestigate this landmark, we conducted a study in four phases:1 Two blindfolded examiners determined the distance betweenthe tuberosity's medial border and the clavicle's lateral endin 100 dried clavicles and then 2 performed subclavian veincannulation in 20 fresh human cadavers using the tuberosityand the suprasternal notch as landmarks. 3 Three-dimensionalreconstructions of the subclavian artery and vein and surroundingstructures were derived from computed tomography datasets of10 patients. The length of the path of a virtual subclavianvein cannulation with the deltoid tuberosity landmark was measuredbilaterally. 4 In a prospective, randomized trial, subclavianvein cannulation was performed in 60 patients with a standardapproach or with the deltoid tuberosity as landmark.

为系统地研究这个解剖标志，作者将本研究分为四个阶段：1 双盲检查100个乾燥的锁骨测量其三角肌粗隆内侧缘至锁骨侧面末端的距离；2 在20具新鲜的尸体上以三角肌粗隆和胸骨上切迹为标志进行锁骨下静脉穿刺；3 利用 CT 所获取的资料资料对10个病人的锁骨下动脉和静脉以及周围结构进行三维重建，测量两侧以三角肌粗隆为标志的虚拟锁骨下静脉置管路径的长度；4 选择60例病人以标准路径或三角肌粗隆为标志进行锁骨下静脉穿刺的前瞻性随机试验研究。

By use of site mutation strategy and PCR technology, we obtained the gene P12X3C that includes full length P1, 2A, 3C and a part of 2B and 3B and the gene P12X3C3D that includes full length P1, 2A, 3C, 3D and a part of 2B and 3B. After being digested by restriction enzyme respectively, the gene P12X3C and the gene P12X3C3D were cloned into the pcDNA3. 1 and pTARGET expression vector that were digested by the same enzyme. Recombinant plasmids were checked by restriction enzyme analysis and nucleic acid sequencing. Further more, recombinant plasmids were transfected into BHK-21 cells by using lipoid. The proteins of foot-and-mouth disease virus , which were expressed in BHK-21 cells, were confirmed by sandwich-ELISA and fluoroscopy, and the capsid of FMDV was tested by electron microscope. In order to evaluate enhanced immune response of guinea pigs against FMDV, DNA vaccines which were designed to produce viral capsids lacking infectious viral nucleic acid and contained the gene P12X3C and the gene P12X3C3D were injected respectively with FMDV 3D protein which was expressed in Pichia Pastoris Secreted expression System and purified or with pcDNA3. 1/IFN which includes the gene IFN-α of cattle. Subsequently, Recombinant plasmids were injected to cattles with or without pcDNA3. 1/IFN. Anti-FMDV antibodies were detected by ELISA, and the T lymphocyte proliferation response was tested by MTT assay, neutralization antibodies titers were analyzed by micro-neutralization assay.

为研制带有O型口蹄疫病毒(Foot-and-Mouth Disease Virus，FMDV)China99株结构蛋白基因及多个非结构蛋白基因的DNA疫苗，本研究通过定点突变方法和PCR扩增方法，获得包含有FMDV China99株结构蛋白P1、非结构蛋白2A、3C以及部分2B、3B编码基因的片段P12X3C和包含有FMDV China99株结构蛋白P1、非结构蛋白2A、3C、3D以及部分2B、3B编码基因的片段P12X3C3D，将获得的基因片段直接/酶切后与同样处理的真核表达质粒连接，分别得到重组质粒pcDNA3.1/P12X3C和pcDNA3.1/P12X3C3D、pTARGET/P12X3C3D；对重组质粒进行序列测定、分析，并将重组质粒分别转染BHK-21细胞，通过双抗体夹心ELISA方法和间接免疫荧光标记方法检测细胞中FMDV抗原的表达，用电子显微镜观察病毒空衣壳的组装；为评价重组质粒作为DNA疫苗对实验动物及本动物的免疫效果，将重组质粒经肌肉注射方法接种豚鼠，并与酵母表达的纯化FMDV China99株3D蛋白及带有牛α干扰素的真核表达质粒pcDNA3.1/IFN分别/同时免疫，第二次免疫后第三周豚鼠攻以1OOID〓或1000ID〓的O型FMDV China99株；随后将质粒pcDNA3.1/P12X3C、pcDNA3.1/P12X3C3D与带有牛α干扰素的真核表达质粒pcDNA3.1/IFN同时免疫牛，三周后经牛舌皮攻以10〓ID〓的O型FMDV China99株。

By use of site mutation strategy and PCR technology, we obtained the gene P12X3C that includes full length PI, 2A, 3C and a part of 2B and 3B and the gene P12X3C3D that includes full length PI, 2A, 3C, 3D and a part of 2B and 3B. After being digested by restriction enzyme respectively, the gene P12X3C and the gene P12X3C3D were cloned into the pcDNA3.1 and pTARGET expression vector that were digested by the same enzyme. Recombinant plasmids were checked by restriction enzyme analysis and nucleic acid sequencing. Further more, recombinant plasmids were transfected into BHK-21 cells by using lipoid. The proteins of foot-and-mouth disease virus, which were expressed in BHK-21 cells, were confirmed by sandwich-ELlSA and fluoroscopy, and the capsid of FMDV was tested by electron microscope. In order to evaluate enhanced immune response of guinea pigs against FMDV, DNA vaccines which were designed to produce viral capsids lacking infectious viral nucleic acid and contained the gene P12X3C and the gene P 12X3C3D were injected respectively with FMDV 3D protein which was expressed in Pichia Pastoris Secreted expression System and purified or with pcDNA3.1/lFN which includes the gene IFN-a of cattle. Subsequently, Recombinant plasmids were injected to catties with or without pcDNA3.1/IFN. Anti-FMDV antibodies were detected by ELISA, and the T lymphocyte proliferation response was tested by MTT assay, neutralization antibodies liters were analyzed by micro-neutralization assay.

为研制带有O型口蹄疫病毒(Foot-and-Mouth Disease Virus，FMDV)China99株结构蛋白基因及多个非结构蛋白基因的DNA疫曲，本研究通过定点突变方法和PCR扩增方法，获得包含有FMDV China99株结构蛋白P1、非结构蛋白2A、3C以及部分2B、3B编码基因的片段P12X3C和包含有FMDV China99株结构蛋白P1、非结构蛋白2A、3C、3D以及部分2B、3B编码基因的片段P12X3C3D，将获得的基因片段直接／酶切后与同样处理的真核表达质粒连接，分别得到重组质粒pcDNA3.1／P12X3C和pcDNA3.1／P12X3C3D、pTARGET／P12X3C3D；对重组质粒进行序列测定、分析，并将重组质粒分别转染BHK-21细胞，通过双抗体夹心ELISA方法和间接免疫荧光标记方法检测细胞中FMDV抗原的表达，用电子显微镜观察病毒空衣壳的组装；为评价重组质粒作为DNA疫苗对实验动物及本动物的免疫效果，将重组质粒经肌肉注射方法接种豚鼠，并与酵母表达的纯化FMDV China99株3D蛋白及带有牛α干扰素的真核表达质粒pcDNA3.1／IFN分别／同时免疫，第二次免疫后第三周豚鼠攻以100ID_(50)或1000ID_(50)的O型FMDV China99株：随后将质粒pcDNA3.1／P12X3C、pcDNA3.1／P12X3C3D与带有牛α干扰素的真核表达质粒pcDNA3.1／IFN同时免疫牛，三周后经牛舌皮攻以10~4ID_(50)的O型FMDV China99株。

The minor core protein s C-encoding gene of Muscovy duck reovirus was cloned into theprokaryotic expression vector pET32a. The recombinant plasmid pET32a-s C was amplified andextracted after being transformed into E.coli DH5a competent cells. Restriction analysis withEcoRⅠand SacⅠand sequences analysis indicated that the recombinant plasmid was inserted withcorrect open reading frame. The fusion protein about 50 ku was produced after induction with 0.15mmol/L IPTG of E.coli competent cells transformed with pET32a-sC. The SDS-PAGE andWestern-bloting test indicated that the fusion protein reacted with the convalescents sera of duckinfected with Muscovy duck reovirus. The indirect ELISA method was developed by using thepurified fusion s C protein as coating antigen. The optimal concentration of s C was 5μg/ml, thedilution of serum sample was 1:40; The results showed that preparation of an ELISA by using sCas coating antigen in detecting 50 field duck sera in comparison with the AGIP were more sensitiveand specific than agar gel immuno-diffusion AGIP test. The results suggest that presence ofantibody against viral protein sC in duck may be a good indicator by the sC-ELISA for detectionof duck infection with reovirus.

同时，本研究将编码外壳蛋白σC的基因克隆于原核表达载体pET32a上，经过EcoRⅠ和SacⅠ双酶切鉴定和序列分析后，得到阳性重组质粒pET32a-σC；将阳性重组质粒pET32a-σC转化到大肠杆菌BL-21感受态细胞中进行诱导表达，经SDS-PAGE和Western-blbtting检测分析，融合表达的蛋白能够与番鸭呼肠孤病毒感染的康复鸭血清发生特异性反应；将融合表达的蛋白纯化后作为包被抗原，建立了检测鸭血清中呼肠孤病毒抗体的间接酶联免疫吸附试验检测方法，此方法中抗原的最佳包被浓度为5μg／ml、标准阳性血清的最适稀释倍数为1：40倍，用此方法对50份鸭血清样品进行检测，并与琼脂糖凝胶扩散试验检测抗体的法相比较，证明此ELISA方法具有良好的特异性和敏感性，本研究为今后鸭呼肠孤病毒诊断试剂盒的研制奠定了基础。

With Death guitarist Schuldiner adopting vocal duties, the band made a major impact on the scene.

随着死亡的吉他手Schuldiner接受主唱的职务，乐队在现实中树立了重要的影响。

But he could still end up breakfasting on Swiss-government issue muesli because all six are accused of nicking around 45 million pounds they should have paid to FIFA.

不过他最后仍有可能沦为瑞士政府&议事餐桌&上的一道早餐，因为这所有六个人都被指控把本应支付给国际足联的大约4500万英镑骗了个精光。