切物的人

Japanese like pursuing the beauty of natural harmony, while they pursue this, they also find the unsolved confrontation between nature and human, and the death is the dividing line, so an appreciation of death,"half sorrow, half beauty" emerged, it was called "pathos", Busido accept this appreciation, and even to perform "hara-kiri" to realize this esthetics notion, under this kind of influence, the estheticism suicide behavior is popular in modern Japanese society, which can be easily found in the Japanese artist suicidal cases, showplace suicide cases and "net-pals group suicide" cases.

日本人是追求自然和谐美的民族，他们在追求此种美的同时，也深深感知到人与大自然不可调和的对立，死亡正是这种对立面的交界，由此派生出一种"亦悲亦美"的死亡审美意识——"物哀"，武士道不仅受此种审美意识影响，更以"切腹"这一自杀行为艺术实践此审美理念，受武士自杀美学的影响，在日本现代社会，唯美式自杀

Communism was obviously an instrument of Soviet domination over conquered societies, but it was also a modus vivendi for large parts of these nations under the conditions in which they were obliged to live. Imre Nagy, the leader of the Hungarian revolt in 1956, and Dubcek became parts of their national legends, which belies the claim that communism was exclusively a foreign imposition.

共产主义明显是苏联统治其所征服的社会的工具，但是它也是这些国家中那些不得不在这一情形下谋杀的大多数人的方式。1956年匈牙利革命领导人纳吉以及杜布切克成为各自国家的象征，这就否定了那种共产主义完全就是外国强加之物的说法。

In order to compare the homology of the gene encoding VP2 between MDPV-Q and MDPV which was registered in the GenBank, also in order to find out changes of the VP2 gene and the immuno-genicity between the wild-strain MDPV-Q and the attenuated strain MDPV-26 which derived by continued passage of virulent wild-type in muscovy duck embryo, a pair of primer (LHMP7/LHMP8) was designed. The upper one LHMP7 and the lower one LHMP8 corresponded to the MDPV-specific nucleotides sequence 2885-2900 and 4618-4604 respectively, according to the MDPV nucleotide sequence databases. The length of the sequence embraced by the primers was 1734bp.

为了获取这一基因片段与国外分离株进行同源性比较，同时也为了了解MDPV强、弱毒株VP2基因之间的异同关系，及其免疫原性的变化规律，通过DNA重组技术，设计了一对引物LHMP7／LHMP8，该对引物选取位于2885～2900及4618～4604的两段序列，跨幅为1734bp，并在这两条引物中分别加入两种限制性核酸内切酶SacⅡ和KpnⅠ的酶切位点，分别对MDPV-Q和由该株病毒经人工致弱后得到的MDPV-26株进行PCR扩增，并将PCR产物克隆到pMD18-T载体上，分别得到二个重组子：pMD18-T—M-Q VP2和pMD18-T-M-26 VP2。

All of the human, rat and mouse CKLF have chemotactic activities on a wide spectrum of leukocytes.

大鼠和小鼠的CKLF同源物，与人CKLF具有类似的剪切形式并且对白细胞有广谱的趋化作用。

The PSCA_3 fragment was selected for its superior expression level in eukaryotic cells.Then the sig-PSCA_3-Fc-GPI genetic fragment was cloned into pVAX1-neo-IRES-GM/B7 vector to construct the final immunological inhanced DNA vaccine pVAX1-PSCA_3-FcGB. Immunofluorescence and flow cytometry were used to confirm the expression of PSCA_3 fragment by transfected into Cos7 cell.Finally,the anti-tumor effect of pVAX1-PSCA_3-FcGB was tested in murine prostate cancer model generated by RM-1 cell line.The animal was immunized with pVAX1-PSCA_3-FcGB DNA vaccine by intramuscular injection plus electroporation,pVAX1 and pVAX1-PSCA_1-FcGB plasmid were used as control.The inhibitory effect of tumor was investigated by observion of forming time,volume and inhibition ratio of tumor.Results:DNA sequencing conformed that the heterological PSCA fusion antigen fragment which was synchronized by overlapping-extending-PCR,was consistent to design.Enzyme digestion analysis showed that the 1 to 4 copies heterological PSCA fusion antigen fragments were constructed successfully.

方法(1)检索GenBank，选择包含人主要T细胞抗原表位序列的人PSCA基因片段，应用异种化抗原设计技术，保留人T细胞抗原表位，设计异种化PSCA融合抗原片段；(2)根据核酸序列按中心模板法设计引物，应用重叠延伸PCR技术拼接合成异种化PSCA融合抗原片段基因，以PCR、限制性酶切和DNA序列测定法进行鉴定：(3)利用DNA限制性内切酶BssHⅡ和MluⅠ酶切后粘端互补的特点，采用同尾酶法构建1—4拷贝异种化PSCA融合抗原片段(PNCA_1-PSCA_4)，并将上述片段分别插入真核表达载体pCI-neo-Fc-GPI中，转染293T细胞，借助免疫荧光+流式细胞术考察插入片段表达效率，最终选定PSCA_3片段进行下一步研究；(4)将sig-PSCA_3-Fc-GPI基因片段自pCI-PSCA_3-Fc-GPI质粒上切下，插入pVAX1-neo-IRES—GM/B7载体中，构建免疫增效DNA疫苗pVAX1-PSCA_3-FcGB，并应用转染Cos7细胞+免疫荧光/流式细胞术方法鉴定其在真核细胞中的表达情况；(5)给8周龄雄性C57BL/6小鼠皮下种植RM-1细胞，制备小鼠前列腺癌模型，并采用股四头肌肌肉注射+电脉冲法(Electroporation，EP)接种DNA疫苗质粒pVAX1-PSCA_3-FcGB，同时接种pVAX1空载体质粒和pVAX1-PSCA_1-FcGB质粒作为对照，通过观察计算免疫动物的成瘤时间、肿瘤体积和抑瘤率，来评价该DNA疫苗在小鼠体内的抑瘤效果。

Another pair of primers including the start codern and the terminer coden were designed to amplify the PAL gene. Two restriction enzyme XbaI and XmaI were added to the two primers respectively, which enable to insert this fragment to the prokaryotic expression vetor pCAMBIA2301-WCS.

在PAL 基因的编码框以启始密码子ATG 和终止密码子TAG 为两端设计一对特异引物，分别在引物5' 端引人XbaI 和XmaI 切位点，用来扩增PAL 基因并与pCAMBIA2301-WCS 构建表达载体pCAMBIA2301-PAL。

Another pair of primers including the start codern and the terminer coden were designed to amplify the PAL gene. Two restriction enzyme XbaI and XmaI were added to the two primers respectively, which enable to insert this fragment to the prokaryotic expression vetor pCAMBIA2301-WCS.

在PAL 基因的编码框以启始密码子ATG 和终止密码子TAG 为两端设计一对特异引物，分别在引物5' 端引人XbaI 和XmaI 酶切位点，用来扩增PAL 基因并与pCAMBIA2301-WCS 构建表达载体pCAMBIA2301-PAL。

Methods: Thirty extracted human molars whose pits and fissures on the occlussal surface had been inoculated with Escherichia coli were randomly divided into three groups: group A was treated with newly confected phenol-formaldehyde resin which resinified the dental fissure contents,group B was sealed Bis-GMA resin in pits and fissures and group C as the control group was only treated by cleanin...

选择30颗人离体磨牙，在咬合面点隙裂沟接种大肠杆菌，随机分为3组；A组以新配制的酚醛树脂使点隙裂沟内容物树脂化，B组以成品Bis-GMA封闭剂作点隙裂沟封闭，C组仅进行表面清洗作为对照组；各组分别沿裂沟点隙处片切牙齿，放入LB培养基培养，观察大肠肝菌生长情况。

Methods: Thirty extracted human molars whose pits and fissures on the occlussal surface had been inoculated with Escherichia coli were randomly divided into three groups: group A was treated with newly confected phenol-formaldehyde resin which resinified the dental fissure contents, group B was sealed Bis-GMA resin in pits and fissures and group C as the control group was only treated by cleaning.

选择30颗人离体磨牙，在咬合面点隙裂沟接种大肠杆菌，随机分为3组；A组以新配制的酚醛树脂使点隙裂沟内容物树脂化，B组以成品Bis-GMA封闭剂作点隙裂沟封闭，C组仅进行表面清洗作为对照组；各组分别沿裂沟点隙处片切牙齿，放入LB培养基培养，观察大肠肝菌生长情况。

You have realise that most of the guys in the newsrooms are basically fans of Man Utd/Liv/Arsenal and find it hard to digest the fact that Chelsea not only are challenging them but also dislodging them.

你可以明显的感觉到现如今新闻编辑部里那些基本都是曼联、利物普或阿森那球迷的人，不得以地接受了切尔西正在挑战他们，并已经撼动他们这一事实（林某人是个好例子，虽然他依然很黑）。

Do you know, i need you to come back

你知道吗，我需要你回来

Yang yinshu、Wang xiangsheng、Li decang,The first discovery of haemaphysalis conicinna.

1〕 杨银书，王祥生，李德昌。安徽省首次发现嗜群血蜱。