核染质

The omp17.3 gene of Brucella abortus was amplified by PCR and then the amplicon was cloned into the eukaryotic expression plasmid pcDNA3.1 to construct a recombinant plasmid pcDNA3.1-omp17.3. Then the recombinant plasmid pcDNA3.1-omp17.3 was transfected into COS7 cells, and the expressed OMP 17.3 was detected by Western-blotting.

采用PCR方法扩增了布鲁氏菌17.3ku外膜蛋白编码基因，并将该基因克隆至真核表达载体pcDNA3.1中，成功构建了真核表达质粒pcDNA3.1-omp17.3.pcDNA3.1-omp17.3转染COS-7细胞后，通过Western-blotting检测到了17.3ku蛋白的瞬时表达。

Methods: Genes of MCP and DAF were connected by 10aa coding sequence of enriching Gly and Ser. Then insert into pcDAN3.1 vector, constituting pcDNA-MD3 expression plasmid of hMCP-DAF fusion gene. The genes were transformed by liposome into the pig endothelial cell; human serum handles masculine cloning, identify the function that restrain breaking of person alexin.

将MCP、DAF两个基因以富含Gly和Ser的10aa编码序列作铰链串接，并接入表达载体pcDNA 3.1中，构成含hMCP-DAF的真核表达质粒(pcDNA-MD3)；以脂质体转染猪内皮细胞，人血清处理阳性克隆细胞，鉴定抑制人补体溶破的功能；应用显微注射将线性基因导入猪受精卵；PCR检测导入基因整合率。

Severance of nerve from muscle will induce retrograde changes in the body of the neuron, known as chromatolysis, which is a sign of dysfunction.

有很多的肌肉神经，会在神经元内减少，就是「色原溶解；核染质溶解；染色质溶解消失；色素溶解消失」也是代表了「官能障碍

Western-blot and immunohistochemical assay methods were employed to detect the expression of FHIT. Apoptosis and cell cycle of cell were analyzed by FCMS. Morphological changes were observed by compound and electron microscope.

然后用流式细胞仪，普通光镜及透射电镜对获得稳定FHIT蛋白表达的转染组细胞和空真核表达质粒转染组细胞及未转染的亲本细胞凋亡的状况进行了考察，另外还通过流式细胞仪检测了三组浙江大学硕士学位论文细胞生长周期的惰况。

The cancer cells are in different differentiation periods: the chromatin of the young cancer cell's nucleus is rich in color and its cytoplasm is basophilous. The young cancer cells don't form into typical glandular cavity; The mature cancer cell is columnar or cubical and its nucleus is located at the base of the cell in gland tube-like arrangement; The decrepit cancer cell stain thin while its nucleus stain dense. The severer's nucleus disintegrate into small fragments. The decrepit cancer cell's arrangement is disorganized, only keeping its glandular shape.

显微镜下癌细胞呈现不同的分化程度：幼稚型癌细胞胞核染色质丰富，胞质嗜碱性，不形成典型腺腔；成熟型癌细胞呈柱状或立方状，细胞核位于细胞基部，呈腺管样排列；衰老型癌细胞胞桨染色变淡，胞核浓染，严重者胞核碎裂成细小碎片，衰老型癌细胞排列紊乱，仅保留腺体样结构轮廓。

Results Lactacystin dealed with C6 glioma cells in 24 h, kytoplasm was concentrated and anachomasised, the chromatin of nucleus was concentrated pyknoticly to see under photo-microscope. The microvilli on surface dispeared、nucleus crenated、chromatin aggregated pyknoticly under transmission electron microscope respectively.

结果 Lactacystin处理体外C6胶质瘤细胞24 h后，光镜下见胞浆浓缩深染，胞核染色质致密浓缩，透射电镜下见细胞表面微绒毛消失，核皱缩，染色质致密浓缩、边集。

Objective Establishment of a hybridoma cell line secreting a monoclonal antibody to facilitate iˉdentification of human tissue kallikrein（KLK1gene,hK1protein）.Methods Mice were immunized with E.coli-exˉpressed GST-hK1fusion protein and their spleen cells were fused with SP2/0myeloblastoma cells.Specificity of the monoclonal antibody was shown by Western blotting and by immunofluorescence.Results The monoclonal antibody reacted specifically to E.coli-expressed hK1and with the KLK1cDNA-transfected COS-1cells.

目的 将人组织激肽释放酶（基因命名 KLK1，酶命名为hK1）cDNA克隆入肠杆菌表达质粒，以重组菌表达的GST-hK1融合蛋白免疫小鼠，获得了分泌hK1特异单克隆抗体的杂交瘤细胞株方法将KLK1cDNA克隆入真核表达质粒，用获得的重组质粒转染COS-1细胞，经间接免疫荧光试验证明，上述单克隆抗体能与转染细胞发生特异反应。

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。

Results: From the second days of the operation, the brain swellen overtly. RBC and phagocyte can be seen in the cerebral interstitial space. The adventitia of the brain arteries and the perivascular Virchow￣Robin space dilated irregularly. The neuronal number of hippocampal CAI sector decreased evidently. In some neuron both the nucleus and cytoplasm condensed, chromatin gathered near the nuclear membrane.

结果： 自术后第二天可见脑组织水肿，红细胞溢出及吞噬细胞浸润，小血管壁外膜及Virchow￣Robin间隙增宽，内含大量水肿液，海马CAI区正常神经元数量明显下降，且排列及不整齐，细胞皱缩，核固缩、深染，核染色质聚集，核周囊膨大，胶质细胞增生。

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。