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It also can be used as alternative antigen of newgeneration vaccine.In this experiment,we screen the major protective antigen hsp65 gene of MAP in order to developnew vaccine especially the DNA vaccine for the prevention of paratuberculosis disease.The hsp65 genewas amplified from MAP C-2 chromosomal DNA by using the PCR technique.We gained a hsp65 gene of 1 626bp.Then PCR product was cloned into pGEM-T vector by T-A clone technique and therecombinant clone was identified by plasmid size,enzyme digestion and PCR identification.The cloneplasmid of pGEM-T- hsp65 was successfully constructed.The nucleotide sequence and deduced aminoacid sequence ofclone gene was analyzed by DNASTAR software.The result indicated that the size ofhsp65 gene consist with M.paratuberculosis K-10 strain in GenBank and the sequential homogeneityreached 99.1%,the amino acid homogeneity reached 99.3%.The preceding analysis indicated that thehsp65 gene was very conservative in M.paratuberculosis.

为了研发预防副结核病的新型疫苗尤其是DNA疫苗及相关蛋白功能，本研究选择了MAP的主要保护性抗原Hsp65蛋白，以副结核分枝杆菌C-2株染色体DNA为模板，以hsp65基因的特异性引物进行PCR扩增，获得了1 626bp的hsp65基因，通过T-A克隆技术，将PCR产物克隆至pGEM-T Vector中，以质粒大小、酶切分析、PCR扩增及序列分析鉴定重组克隆，成功地构建出克隆质粒pGEM-T-hsp65，以DNASTAR软件分析了所克隆基因的核苷酸序列和推导的氨基酸序列，结果表明，所获得的hsp65基因与GenBank中MAPK-10株该基因核苷酸大小完全一致，两者核苷酸序列的同源性为99.1％，氨基酸序列的同源性为99.3％，表明该基因在副结核分枝杆菌中高度保守。

Experimental results show that,compared with full redundancy which replicates full error flow graph,partial redundancy can reduce register usage by 6.25%, reduce power dissipation by over 17%,reduces total execution cycles by nearly 26%,and improves performance by over 22%,at the cost of 6.25% lower nodes coverage.

实验显示，与复制全部错误流图的完全冗余相比，在结点覆盖率降低6.25%的情况下，部分冗余算法最多能够减少寄存器的使用数量6.25%，减少功耗超过17%，减少执行时间接近26%，同时提高性能超过22%。

The Lord tried him by delaying to fulfill His promise. Satan tried him by temptation; men tried him by jealousy, distrust, and opposition; Sarah tried him by her peevishness.

神用延迟应许的方法试验他；撒但用诱惑试验他；世人用嫉妒、怀疑、反对，试验他；撒拉用她乖戾的性情试验他。

The judgment of the general term and that entered upon the report of the referee should therefore be reversed, and judgment should be entered as follows: That Elmer E. Palmer and the administrator be enjoined from using any of the personalty or real estate left by the testator for Elmer's benefit; that the devise and bequest in the will to Elmer be declared ineffective to pass the title to him; that by reason of the crime of murder committed upon the grandfather he is deprived of any interest in the estate left by him; that the plaintiffs are the true owners of the real and personal estate left by the testator, subject to the charge in favor of Elmer's mother and the widow of the testator, under the antenuptial agreement, and that the plaintiffs have costs in all the courts against Elmer.

原审普通法院的判决以及法官的审判报告应当被撤销，作出如下判决：埃尔默·帕尔默和遗产管理人不能动用遗嘱人为埃尔默遗增的任何财产；遗嘱中赠与埃尔默的动产和不动产不发生有效转移；谋杀者埃尔默因其犯罪行为被剥夺获得遗产的权利；两名原告是遗嘱人动产和不动产的真正继承人，但应由埃尔默的母亲和遗嘱人的遗孀依据婚前协议来照管，埃尔默承担两原告支付的所有诉讼费。

Finally this thesis analyses the efficient revocable group signature scheme with forward security proposed by Chen Shaozhen and Li Daxirig. An adversary can personate a legal group member by forge a group membership certificate, and then can forge group signatures that can be verified by a verifier. And the group manager can also forge group signatures that can be verified by a verifier. So the scheme is insecure. Meanwhile, there are redundancy steps in the scheme, thus the efficiency is low.

接着，对陈少真和李大兴提出的一种有效取消的前向安全群签名方案进行了密码学分析，一方面，攻击者可以通过伪造群成员证书来伪造能够通过验证的群签名；同时，群管理员也可以伪造一个可以通过验证的群签名，因此，该群签名体制是不安全的；另一方面，该体制存在冗余数据。

About a Year and half after I had entertain'd these Notions, and by long musing, had as it were resolved them all into nothing, for want of an Occasion to put them in Execution, I was surpriz'd one Morning early, with seeing no less than five Canoes all on Shore together on my side the Island; and the People who belong'd to them all landed, and out of my sight: The Number of them broke all my Measures, for seeing so many, and knowing that they always came four or six, or sometimes more in a Boat, I could not tell what to think of it, Or how to take my Measures, to attack Twenty or Thirty Men single handed; so I lay still in my Castle, perplex'd and discomforted: However I put my self into all the same Postures for an Attack that I had formerly provided, and was just ready for Action, if any Thing had presented; having waited a good while, listening to hear if they made any Noise; at length being very impatient, I set my Guns at the Foot of my Ladder, and clamber'd up to the Top of the Hill, by my two Stages as usual; standing so however that my Head did not appear above the Hill, so that they could not perceive me by any Means; here I observ'd by the help of my Perspective Glass, that they were no less than Thirty in Number, that they had a Fire kindled, that they had had Meat dress'd.

我自从有了这些想法之后，平时就经常会想到这件事，可是因为没有机会付诸实施，因此一直都毫无结果。这样大约又过了一年半光景。一天清晨，我忽然发现有五只独木舟在岛这头靠了岸，船上的人都已上了岛，但却不知道他们去哪儿了。他们来的人这么多，把我的计划彻底打破了。因为我知道，一只独木舟一般载五、六个人，有时甚至更多。现在一下子来了这么多船，少说他有二三十人，我一个人单枪匹马，如何能对付他们呢！因此，我只好悄悄躲到城堡里去，坐立不安，一筹莫展。可是，我还是根据过去的计划，进行作战准备，以便一有机会，立即行动。我等了好久，留神听他们的动静，最后，实在耐不住了，就把枪放在梯子脚下，像平时那样，分作两步爬上小山顶。我站在那里，尽量不把头露出来，唯恐被他们看见。我拿起望远镜进行观察，发现他们不下三十人，并且已经生起了火，正在煮肉。至于他们怎样煮的，煮的又究竟是什么肉，我就不得而知了。

Methods: Human leukemia cell line K562 was treated with different concentrations of WM. The proliferation of K562 cells was examined by MTT assay. DNA damage in K562 cells was examined by single cell gel electrophoresis assay, and apoptosis of K562 cells was detected by Annexin V-FITC/PI double staining. The expressions of total Akt, phosphorate Akt, and NF-κB p65 mRNA and protein were detected by RT-PCR and Western blotting, respectively.

以不同浓度的渥曼青霉素作用于人类髓细胞白血病细胞K562，采用MTT法检测细胞增殖活性，单细胞凝胶电泳技术检测细胞DNA损伤形成的"彗星"样拖尾现象，Annexin V FITC/PI双标法检测细胞凋亡，Western blotting、RT-PCR检测渥曼青霉素作用K562细胞后总Akt和磷酸化Akt以及NF-κB基因及蛋白表达水平的变化。

The test was divided into three groups randomly as yh-16 - experiment group, pbs- blank control group and rpmi-1640 medium - positive group.③in the following 24h、48h and 72h interventions, the morphous of ec was observed by photology inverted microscope, the cytoactive was detected by method of mtt and the cell cycle was investigated by flow cytometry.the expression of vegf was dectected by real-time fluorescent quantitate polymerase chain reaction(real-time pcr).

以有效浓度的yh-16作用于ec，在不同时相点光镜下观察ec形态，mtt检测细胞活性，流式细胞术检测细胞周期，实时荧光定量pcr检测vegf的表达量。

Results: Among 32 tested MNs, depolarizing responses(excitatory postsynaptic potential, EPSP) were elicited by cVLF stimulation in 21 MNs, hyperpolarizing response (inhibitory postsynaptic potential, IPSP) in 1 MN, and IPSP preceded by EPSP in 4 MNs. The cVLFEPSPs were stimulus intensity-dependent, and could be abolished by low Ca(superscript 2+)/high Mg(superscript 2+) solution. compared with iVLF-EPSPs, cVLF-EPSPs showed longer latency (P.001). The cVLF-IPSP presented with membrane potential-dependent property and was eliminated by the perfusion of picrotoxin (30 μmol/L) and strychnine (1.0 μmol/L).

结果：在32个测试的MNs，观察到cVLF电刺激可在21个MNs上诱发去极化反应（即cVLF性兴奋性突触后电位，cVLF-EPSP），在1个MN上诱发超极化反应（即cVLF性抑制性突触后电位，cVLF-IPSP），在4个MNs上诱发cVLF-EPSP后复合有cVLF-IPSP的反应。cVLF-EPSP具有刺激强度依赖性、被低钙高镁溶液取消的特性，与i VLF性EPSP相比，有潜伏期较长的特点(P.001)。cVLF-IPSP呈膜电位依赖性，并被印防己毒素(30μmol/L)及士的宁(1.0μmol/L)取消。

A:Group1,widely opened by 20 μg·kg-1 of AD;B:Group 2,moderately opened by 10μg·kg-1 of AD;:Group 3,Slightly opened by 5μg·kg-1 of AD;D:Group 4,except a non-BBB area,pineal body,BBB was not opened in adjacent brain area by 3μg·kg-1 of AD.

a：第一组，AD 20 μg·kg-1引起的广泛开放；B：第二组，AD10 μg·kg-1引起的中度开放；C：AD 5 μg·kg-1引起的轻度开放；D：除缺乏BBB的松果体之外，AD3 μg·kg-1未引起其它脑区的BBB开放

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