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Methods:The total RNA was extracted from mouse RAW264.7 cells stimulated by LPS.The sequence including the whole length of HMGB1 was amplified by RT-PCR and inserted into pMD-19T.The combinant vector was used as a template for PCR which was cloned into vector pMD-19T,then subcloned into expression vector pET-26b with pelB signal sequence and His-Taq sequence.After transforming E.coli BL21(DE3) and four hours induction by IPTG,HMGB1 expression confirmed by SDS-PAGE and the purification was performed by Ni2+-chelate affinity chromatograph.

脂多糖刺激后的RAW264.7细胞，提取总RNA，经RT-PCR扩增出含HMGB1的目的片段，克隆于pMD-19T载体，再亚克隆至含有pelB引导肽及His-标签肽的高效表达载体pET-26b，转化大肠杆菌BL21（DE3），经IPTG诱导后行SDS-PAGE鉴定目标蛋白表达，用镍鳌合琼脂糖凝胶亲和层析法分离纯化含His-标签肽的目的蛋白。

Based on the settings and geological and geochemical characteristics of granitoids of different orogenic belts (exemplified by orogenic granitoids in Altay, East Kunlun, Yanshan in North China, Northeast China and Nanling in South China), the relationships between granitoids and continental crustal growth are discussed, and five modes of continental crustal growth in the continent of China are proposed;they are as follows:(1)the Altay mode of continental growth is that the material and heat of the Paleozoic convertive mantle formed on the background of the Paleo-Asian Ocean were input into the Paleozoic continent and mixed with the materials of the upper crust;(2) the East Kunlun mode is that the basement of the Proterozoic orogenic belt was reworked by the heat input and materials of the Paleozoic-early Mesozoic convective mantle formed on the background of TTG continental crust of the Proterozoic orogenic belt;(3) the Northeast China style is that the Phanerozoic continental crust was reworked by the heat input and materials from the convective mantle formed on the background of the Yanshanian Central Asian orogenic belt;(4) the Yanshan mode is that the Archean basement was reworked by the heat input and materials from the Yanshanian convective mantle;and (5) the Nanling mode is that in South China what was input by the Yanshanian convective mantle into the continent was mainly heat and subordinately materials and the crustal material recycle was the dominant mode for the continental growth (showing zero continental growth).

根据不同造山带花岗岩的形成背景，地质地球化学特征差异，以阿尔泰，东昆仑，华北燕山，东北和南岭造山带花岗岩为例讨论花岗岩与大陆地壳生长的关系，区分出中国大陆的5种大陆地壳生长方式：阿尔泰式是古亚洲洋背景上形成的古生代对流地幔物质，热输入和上地壳混合为主的方式；东昆仑式是元古代造山带TTG陆壳背景基础上古生代—早中生代对流地幔物质和热输入，改造元古宙造山带基底的方式；东北式是燕山期中亚造山带背景上对流地幔物质和热输入改造显生宙陆壳的生长方式；燕山式是燕山期对流地幔物质和热输入改造太古宙基底的方式；南岭式燕山期对流地幔输入大陆的是以热为主，物质为辅，大陆地壳生长是以陆壳物质再循环为主的生长方式。

The equivalent standard pollution loadings were compared among different rigions, the results indicated that the water pollution threaten caused by agricultural non-point source pollution was more serious in the south of Jiangsu province and in Nantong than in the other rigions;The equivalentStandard pollution loading rates of different agricultural non-point source pollution in different regions were analysised by clustering analysis method,and the results indicated that Nanjing、 Wuxi、 Zhenjiang and Changzhou were polluted mainly by domestic living pollution;Xuzhou、 Suqian、 Taizhou、 Yancheng、 Lianyungang、 Huaian and Nantong were polluted mainly by chemical fertilizer pollution; Suzhou and Yangzhou were polluted corresponsively by animals feces pollution 、 domestic living pollution、 chemical fertilizer pollution and fish breeding pond pollution.

对各地区的等标污染指数进行比较，发现农业非点源污染对水环境造成的威胁苏南地区和南通明显高于其它苏中、苏北地区；对各个市各非点源污染源的污染负荷率进行聚类分析，发现南京、无锡、镇江、常州以生活污染为主；徐州、宿迁、泰州、盐城、连云港、淮安、南通以化肥污染为主；苏州、扬州两地畜禽粪便污染、生活污染、化肥污染和精养鱼塘污染均占一定的比例。

Through block wall experimental study we can find out that cracks would appear late and the load of crack-resisting and limit-resisting are increased obviously by the constructional measures in the wall. It is very effective method that the splayed cracks are controlled by sill ring beam, the horizontal bending cracks are controlled by structural column on the side of the wall and the horizontal cracks on the top of the wall are controlled by the whole ring beam. It is the most effective method that wall cracks are controlled by structural column, crout column and sill ring beam.

通过试验主要得到在墙体中采取构造措施后，墙体裂缝推迟出现，开裂荷载和极限荷载都有很大程度的提高，同时发现窗台圈梁在控制窗角的八字缝、墙体两侧构造柱对于防止水平弯曲缝以及全截面的圈梁在防止墙体和圈梁交界处的裂缝的出现能起到很好的效果，通过试验确定在砌块墙体中采取&构造柱+芯柱+窗台圈梁&的构造措施来控制裂缝是最为经济合理有效的。

Results 15 eyes of peripheral retinal dry hole and peripheral retinal degeneration were treated only by laser treatment and successful rate was 100%;10 eyes with local rhegmatogenous retinal detachment,were initially treated with herbal dehydrant,followed by laser treatment and successful rate was 80%;2 eyes failed due to the traction of vitrous were successfully treated with the aids of local pad;141 eyes with retinal detachment were treated by scleral buckling combined with krypton laser and success rate was 96.5%,5 eyes failed due to the traction of vitrous were successfully treated with the aids of vetroretional operation;27 eyes with complicated retinal detachment were treated by vetroretional operation,followed by intro????and post????

结果 视网膜周边干性裂孔和视网膜周边变性单纯用氪多波长激光治疗15眼，成功100%；局限性孔源性视网膜浅脱离10眼，先使用脱水剂、卧床休息、单眼加压包扎等措施，而后行激光治疗，成功率80%，2眼因玻璃体有牵引结合局部垫压术后成功；巩膜外扣带术后激光封孔141眼，成功率96.5%。5眼因玻璃体有牵引行玻璃体手术后眼内激光封闭裂孔成功；复杂性视网膜脱离经玻璃体切割手术，术中及术后激光封闭裂孔27眼，成功100%。

Results 15 eyes of peripheral retinal dry hole and peripheral retinal degeneration were treated only by laser treatment and successful rate was 100%;10 eyes with local rhegmatogenous retinal detachment,were initially treated with herbal dehydrant,followed by laser treatment and successful rate was 80%;2 eyes failed due to the traction of vitrous were successfully treated with the aids of local pad;141 eyes with retinal detachment were treated by scleral buckling combined with krypton laser and success rate was 96.5%,5 eyes failed due to the traction of vitrous were successfully treated with the aids of vetroretional operation;27 eyes with complicated retinal detachment were treated by vetroretional operation,followed by intro-and post-operation,and success rate 100%.

结果 视网膜周边干性裂孔和视网膜周边变性单纯用氪多波长激光治疗15眼，成功100%；局限性孔源性视网膜浅脱离10眼，先使用脱水剂、卧床休息、单眼加压包扎等措施，而后行激光治疗，成功率80%，2眼因玻璃体有牵引结合局部垫压术后成功；巩膜外扣带术后激光封孔141眼，成功率96.5%。5眼因玻璃体有牵引行玻璃体手术后眼内激光封闭裂孔成功；复杂性视网膜脱离经玻璃体切割手术，术中及术后激光封闭裂孔27眼，成功100%。

Results: Tyroserleutide can significantly increase the life span of H22 tumor-bearing mice by 50-70% in dosages of 20ug/kg/d-80ug/kg/d,specially the high dosage of 80ug/ml can significantly increase the life span by 69.24%; Tyroserleutide can inhibit the growth of transplanted hepatocellular tumor BEL-7402 in nude mice,the rate of tumor inhibition was25-50% in dosages of 40-320ug/ml ,the inhibition rate of 160ng/ml was 44.03%; Tyroserleutide could inhibit the growth of H22 and BEL-7402 tumor in a dose-dependent manner. Simultaneously, tumoricidal activity of tyroserleutide against BEL-7402 cell line in vitro was observed hinger when compared with the control group(P.05).The inhibition effect of 72hrs was higher than 24hrs,48hrs,96hrs.And specially the high dosage of 160ug/ml can significantly inhibit growth of tumor cell by 19.36%. Tyroserleutide can activated PEM and marked enhance cytotoxicity andphagocytosis functions in vitro and in vivo. The OD values of cytotoxicity were observed hinger when compared with the control group(P.05).The cytotoxicity of macrophages activated by tyroserleutide against BEL-7402 and B16-F10 was 35.58%,61.2% in vitro and21.39%,47.63% in vivo. The cytotoxicity rate of nude mice PEM was 32.86%,73.07% in vivo. Furthermore, tyroserleutide alone could stimulated the production of IL-1B TNF- a and NO by M . Tyroserleutide and LPS could synergistically activated M producing more cytotoxicity effectors. Conclusion: Tyroserleutide had inhibition functions against hepatoma carcinoma .Its possible mechanisms were related to the affect that Tyroserleutide could inhibit tumor cell directively and induce tumor cells apoptosis or death effectively.

结果：酪丝亮肽能显著延长腹水型肝癌H_(22)小鼠的生存时间，给药剂量为80μg／kg／d时疗效最显著，达到69.24％，在20μg／kg／d-80μg／kg／d剂量范围内生命延长率为50-70％，给药剂量与荷瘤鼠生存时间呈现一定量效关系；酪丝亮肽能显著抑制人肝癌BEL-7402移植瘤裸鼠的肿瘤生长，给药剂量为160μg／kg／d时疗效最显著，抑制率为44.03％，并且在40-320μg／kg／d剂量范围内抑制率为25-50％，给药剂量与肿瘤抑制率呈现一定量效关系；酪丝亮肽体外对人肝癌BEL-7402细胞生长有一定的抑制作用，在作用72hrs时各浓度酪丝亮肽对肿瘤细胞的抑制作用较24hrs、48hrs、96hrs明显，其中浓度为100μg／ml时抑制率达19.36％；酪丝亮肽体内外均能增强小鼠腹腔巨噬细胞对肿瘤细胞的杀伤：体外作用中巨噬细胞对BEL-7402、B16-F10的杀伤功能明显增强，与效应细胞对照组相比有显著性差异(P＜0.05)杀伤率分别达到35.58％、61.2％；体内作用中巨噬细胞对BEL-7402、B16-F10的杀伤功能明显增强，与生理盐水对照组相比有显著性差异(P 。05)，杀伤率分别达到21.39%、47.63%；裸鼠腹腔巨噬细胞经酪丝亮肤作用后对BEL一7402、B 16一F10杀伤功能明显增强，与生理盐水对照组相比有显著性差异(P.05)，最高杀伤率分别达到32.86%、73.07%；酪丝亮肤能增强单核巨噬细胞系统的吞噬功能，吞噬指数与生理盐水组比较有显著性差异(P.05)；酪丝亮肤体外作用能促进小鼠腹腔巨噬细胞分泌合成细胞毒效应分子IL一lp、TNF一Q和NO，与效应细胞对照组相比有显著性差异(P.05)；酪丝亮肤体内作用能促进小鼠腹腔巨噬细胞分泌合成细胞毒效应分子IL一lp、TNF一Q和NO，与生理盐水对照组相比有显著性差异(P.05)；酪丝亮肤能促进鼠巨噬细胞株R戌W264.7分泌合成IL一1p和NO，IL一1日、NO水平分别在酪丝亮肤作用24hrs、12hrs时达到高峰，酪丝亮肤单独应用能提高巨噬细胞的分泌合成功能，而且酪丝亮肤能与LPS协同作用刺激巨噬细胞的细胞毒效应分子分泌合成。

Our homestate operation, led by Rod Eldred; our workers' compensation business, headed by Brad Kinstler; our credit card operation, managed by the Kizer family; National Indemnity's traditional auto and general liability business, led by Don Wurster - all of these generated significant underwriting profits accompanied by substantial float. We can conclude this section as we did last year: All in all, we have a first-class insurance business.

伯克希尔其它保险事业 1994 年的营运表现皆相当杰出，由 Rod 领导的住宅保险， Brad 带领的员工退休保险以及由 Kizer 家族经营的信用卡保险业务，以及由 Don 所领导的国家产险所从事的传统汽车保险与一般责任险方面，总的来说，这四类业务都表现的相当不错，不但有承保获利，还贡献了相当大金额的保险浮存金。

Berkshire's other insurance operations had excellent results in 1994. Our homestate operation, led by Rod Eldred; our workers' compensation business, headed by Brad Kinstler; our credit card operation, managed by the Kizer family; National Indemnity's traditional auto and general liability business, led by Don Wurster - all of these generated significant underwriting profits accompanied by substantial float.

Berkshire其它保险事业1994年的营运表现皆相当杰出，由Rod领导的住宅保险、Brad带领的员工退休保险以及由Kizer家族经营的信用卡保险业务，以及由Don所领导的国家产险所从事的传统汽车保险与一般责任险方面，总的来说，这四类业务都表现的相当不错，不但有承保获利，还贡献了相当大金额的保险浮存金。。

Product fault ink UN-uniform UN-uniform ink products were:(1) water and ink, ink and wash painting the rubber roll version of wear, spindle bearing wear, cots epiderm aging decline, ink, etc;(2) series water roll of layer debranning, cylindricity wear, a thin layer of shoulderpieces for intergradle monlchamus, a product of long a portion of the Ribbon, resulting in a roller 跞 movement oscillating anisotropism;(3) support by Edition of water, rollers, the cylinder, swayig stand, wear, resulting in version tiaodong Cot by cause pressure by version is unstable, produce ink imbalance, ink, ink plate, staves, relying on the old version (students'overloaded) version of COTS and pressure;(4) the water roll chemiluminscent set of aging or poor quality water decline caused by transmission;(5) does not have a regular check of the drum-and-ink roller and version by version-and-ink roller and pressures;(6) if the drum gear wear, and gear spacing is the same as the regular ink "" gear.

二、产物墨色不平均障碍产物墨色不平均的来因有：（1）传水、传墨、匀墨和靠版水墨各橡胶辊的圆柱度磨损超不差，拆轴承的轴脚磨损，胶辊不内脏嫩化，传墨本领回升等；（2）串水辊度层脱皮，圆柱度磨损超不差，暗地有薄积墨层，两脚长出局部有积墨带，变不败滚子跞不静串墨不不平衡；（3）撑持靠版水、墨辊的锁脚，摆不静支架，磨损惨重，变不败胶辊靠版时跳不静致使靠版不抬辛不安定，爆发水墨不不平衡，呈现墨色不不平衡、干版、水墨杠、嫩版等本体；（4）水辊绒套嫩化或质度不差变不败传水本领回升；（5）操纵按期校核各水墨辊及版滚筒与靠版水墨辊的不抬辛；（6）若滚筒传不静齿轮磨损超不差，呈现与齿轮间距不异的有次序的&齿轮墨杠&。不一、套印障碍 1。

They weren't aggressive, but I yelled and threw a rock in their direction to get them off the trail and away from me, just in case.

他们没有侵略性，但我大喊，并在他们的方向扔石头让他们过的线索，远离我，以防万一。

In slot 2 in your bag put wrapping paper, quantity does not matter in this case.

在你的书包里槽2把包装纸、数量无关紧要。