动物细胞

The animal pole process punched in to the follicle and made a gap on it, and then the whole oocyte got out through this gap. Two forces may response to ovulation: the hydration of jelly space and contraction of follicle. Two MTOC are anchored beneath the animal pole process.

卵母细胞通过动物极突起连接到滤泡上，在脱滤泡作用开始时，卵母细胞动物极突起刺入并穿过滤泡膜，使得滤泡膜表面形成一个缺口，然后整个卵母细胞开始从缺口处挤出。

Five models were used to investigate the "Changdong"" s anti-tumor effect: mice S180 sarcoma solid model, mice liver cancer H22 solid model, mice Lewis lung cancer model, as well as human liver cancer QGY and colon cancer LOVO implanted in naked mice; mice liver cancer H22 was also used to evaluate the enhancive effect of "Changdong" when it was co-applied with other anti-tumor agent or with radiotherapy; The "Changdong"" s cytotoxic effect in vitro on such tumors as QGY, LOVO, lung cancer A549, breast cancer MCF-7, as well as mice leukemia P388 was observed through MTT method. And investigate the effect of "Changdong" on proliferation of spleen lymphocyte, on activity of NK cell and on synthesis of IL-2 in mice bearing Lewis lung cancer, respectively. With propidium iodide straining, cell aigptosis and cell circle were analyzed by flow cytometry. Result:"Changdong" has an evident tumor inhibitive effect on mice tumor model as well as human tumor implanted in naked mice with a quantity-effect relationship.

使用小鼠S180肉瘤、小鼠肝癌H22实体瘤和小鼠Lewis肺癌模型，以及人体肝癌QGY和人体肠癌LOVO裸鼠异种移植模型，进行"长动"的抗肿瘤药效学研究：用小鼠肝癌H22进行"长动"合并放疗和化疗的增效作用研究；用MTT法进行"长动"对肝癌QGY、肠癌LOVO、肺癌A549、乳腺癌MCF-7、小鼠白血病P388等10株人体和动物肿瘤细胞的体外细胞毒性作用研究；用淋巴细胞转化试验、小鼠NK细胞活性试验、小鼠胸腺细胞增殖法分别测定"长动"对荷Lewis肺癌小鼠的脾淋巴细胞增殖的影响、自然杀伤细胞活性的影响和对小鼠产生白介素-2(IL-2)的影响；用流式细胞仪PI单染色法进行"长动"对人体肝癌QGY细胞的体外诱导凋亡作用和对肿瘤细胞周期的影响的研究。

Under transmission electron microscope, cellular swelling in brain tissue, nuclear membrane of neuron has incisures, some chromatic agglutination, group A mainly appears endoplasmic reticulum distention and glial cell endocytosis, while group B mainly appears histolysis; cardiac fibrous structure is chaotic, Z line concentrates, mitochondria swelling, nucleus deform; the ecphyma of podocyte becomes thinner and longer like villus, some mix together, plasma membrane infolding of renal cells decreases, mitochondria malformation, cells in lumens.

透射电镜观察显示，试验组动物脑组织细胞肿胀，神经元核膜出现切迹，部分染色质凝集，A组动物出现内质网扩张及胶质细胞吞噬现象，B组动物以组织溶解为主；心肌纤维结构紊乱，Z线聚集，收缩带形成，线粒体肿胀，核变形；肾小球足突细长，绒毛化，并可见足突融合，肾小管上皮细胞质膜内褶减少，线粒体畸形，管腔可见脱落细胞，上述改变均以B组为著。

In the present study,we intend to establish a new model of traumatic brain-steminjury,and to study the pathological changes of neuron,axon,astrocyte,plasmaproteins,blood vessel endothelia,synapse,neurotransmitter and cytokines in rat'sand human injured brain-stem,using routing histochemistry,immunohistochemistry,in situ hybridization,LSCM and computerized image analysis.

综合现有文献，本研究拟建立新的脑干损伤动物模型，并采用常规病理组化、免疫组化、原位杂交、LSCM以及计算机图像分析等技术，对动物及人体脑干组织中的神经元、星形胶质细胞、血浆蛋白及血管内皮细胞，以及轴索、突触、神经递质及细胞因子等进行研究，并对脑干损伤的致死性病变进行初步定位，以确立对脑干损伤的鉴定和推断脑干损伤时间有意义的客观指标。

Get high purity DCs by Cultured plastic-adherent monocytes isolated from healthy human peripheral blood with GM-CSF and IL-4 for 7 days. To observe the morphology of DCs by inverted phase contrast microscope ,electron microscope and laser confocal microscope. Analyse phenotype of DCs with flow cytometry. Investigate the endocytosis ability of DCs as a group by Horseradish peroxidase endocytosis assay. To appraise allogeneic mixed lymphocytes reaction of DCs by MTT reduction assay. Analyse the levels of IL-12 and TNF in liquids of cultured medium by ELISA and MTT reduction assay respectively. Soluble antigens of HCCs was obtained by 3 freeze-and-thaw cycles. Biological characteristics of HC soluble antigens pulsed DCs were monitored by flow cytometry. According to MTT reduction assay estimated the cell proliferation of self lymphocytes activated by HC antigens pulsed DCs. Get high purity BCG HSP 70 protein by SDS-PAGE electrophoresis and determined its biological activity with ELISA. Analyse phenotype of antigen pulsed DCs primed by BCG HSP70 with flow cytometry. By MTT reduction assay estimated the cell proliferation of self lymphocytes and the MLR of DC based vaccine. Analyse expression of HLA-DR molecule on surface of HCC lines. The IFN-γ mRNA in lymphocytes after actived by DC vaccine and the Fas-L expression on DC and DC vaccine primed lymphocytes were detected by in situ hybridization and flow cytometry respectively. Specific cytotoxity lysis of T lymphocytes and nonspecific inhibition of liquids in culture medium against HCC lines were also tested. Detect expression of hAFP on four HCC lines with Cell-ELISA. Induce apoptosis of HCCs with actinomycin-D. Interaction of DCs and apoptotic cells was observed under transmission electron microscope. Growth inhibition test of DC against HCC lines was also performed. Establish the nude mouse model bearing human HC xenografts and indentify the characteristic of tumour by histochemistry and immunohistochemistry techniques. Prevent and treat transplanted human HC on nude mouse with Freezing and anabiotic HC specific lymphocytes.

用GM-CSF和IL-4从健康人外周血诱导DC；分别用倒置相差显微镜、电子显微镜及激光共聚焦显微镜观察DC形态；流式细胞术检测DC表型；HRP吞噬实验测定DC的群体内吞能力；MTT法检测同种异体混合淋巴细胞反应；ELISA法和MTT法分别测定DC培养上清液中IL-12和TNF水平；冻融法制备肝癌细胞可溶性抗原；流式细胞术检测负载肝癌可溶性抗原后DC的生物学特性；MTT法检测DC负载肝癌抗原后对自身淋巴细胞增殖的影响；SDS-PAGE制备电泳纯化BCG HSP70并鉴定纯度，ELISA测定活性；流式细胞术检测负载抗原DC经BCGHSP 70活化后的表型；MTT法检测肝癌DC疫苗对自身淋巴细胞增殖的影响和混合淋巴细胞反应；流式细胞术检测肝癌细胞表面HLA-DR表达；MTT法检测肝癌DC疫苗对自身淋巴细胞的活化；原位杂交法检测肝癌DC疫苗活化后的淋巴细胞IFN-γmRNA表达；流式细胞术检测DC和肝癌DC疫苗活化后淋巴细胞表面Fas-L；MTT法分别检测肝癌DC疫苗活化的淋巴细胞和其培养上清对肝癌细胞的特异性杀伤和非特异性抑制作用；Cell-ELISA检测人肝癌细胞hAFP表达；MTT法检测负载AFP表位肽和凋亡肝癌细胞DC对自身淋巴细胞增殖的影响；ELISA法和MTT法分别测定活化后淋巴细胞培养上清中TNF和IL-12水平；肝癌细胞凋亡的诱导和检测；DC吞噬凋亡肝癌细胞后的电子显微镜观察；DC对肝癌细胞的生长抑制试验；人肝癌裸鼠皮下移植瘤动物模型的建立及其组织学和免疫组织化学鉴定；DC及肝癌特异性淋巴细胞预防和治疗人肝癌裸鼠皮下移植瘤；冻存和复苏后的肝癌特异性淋巴细胞预防和治疗人肝癌裸鼠皮下移植瘤。

Results In animal study, lumbricus could inhibit the growth of staphylococci, bacillus coli and bacillus aeruginosus. The time of wound healing in experimental group was 4 days shorter than that in control group. At 4d and 7 day the numbers of the capillary, blood vessel endodermis and desmohemoblast desmocyte and splitting epithelium of trial group were much more than those of control group. At 4d the trial group′s numbers of splitting mesenchymal cell were much more than that of control group. From 3d on the wound healing and granulation filling of experimental group were much quicker than those of control group.

结果 动物实验中，地龙能抑制金黄色葡萄球菌、大肠杆菌及绿脓杆菌的生长；实验组创面愈合时间较对照组提前了4天；第4、7天实验组动物创面的毛细血管数、血管腔内皮细胞数及间质成纤维细胞数均较对照组明显增多，上皮细胞分裂象也高于对照组，第4天的间皮细胞分裂象也高于对照组；自第3天始创面愈合及肉芽充填速度实验组明显快于对照组。

The mice inoculated with Coxiella burnetii intrana- sally developed interstitial pneumonia,while the primary pathological changes of mice inoculated intraperitoneally are granulomas in spleen and liver.2.The pathological changes became more severe followed the dosage increasing.3.Coxiella burnetii can be detected in spleen and liver at day 2 after inoculation.the lesion became more and more serious from day 2 to day 12.The characteristic changes were observed at day 7,and recovered at day 14. 4.The reticuloendothelial system are main target of Coxiella burnetii.The pathogen was detected in cytoplasm of monocyte -macrophages of spleen, liver, lung, and endothelioid cells of blood vessel. 5. Coxiella burnetii can be found in macrophages lysosomes by electron microscopy. Most of them are round or rod, and polymorphic shape can also be observed in different size.

结果：1、通过不同感染途径的实验证实，滴鼻感染的小鼠主要表现为间质性肺炎，而腹腔注射感染小鼠则以脾脏、肝脏肉芽肿为主要病变。2、通过不同剂量的感染实验发现，随着感染Q热立克次体剂量的加大，动物病变愈加严重。3、通过感染后不同时间的动态病理学观察发现，在腹腔注射后第2d的脾和肝脏即可发现病原体，主要脏器的病理变化从第2d到第12d逐渐加重，第7d动物的病变最典型，至感染后14d动物的受损器官已开始出现修复性变化。4、 Q热立克次体主要侵害机体的网状内皮系统，在感染小鼠的肝、脾、肺和外周血管单核巨噬细胞以及血管内皮细胞胞浆中查见病原体。5、透射电镜观察可见Q热立克次体主要位于巨噬细胞吞噬溶酶体内，呈多形性，多见圆形和杆状，大小不一。

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。

Since an excessive cell growth is considered as the main pathological event, cell proliferation model occupy most of the animal models, and these cells include retinal pigment epithelial cell, fibroblast cell, cartilage cell and vascular endothelial cell.

由于PVR的主要病理变化是细胞的过度增生，因而动物模型多以细胞增生模型为主，细胞种类有视网膜色素上皮细胞、成纤维细胞、软骨细胞、血管内皮细胞等。

Oocyte samples from one group were collected to detect the presence and integration of HBV DNA within cells and chromosomes using PCR, Southern blot, dot hybridization and fluorescence in situ hybridization. The female animals from another group were mated with their normal males, respectively. Their zygotes, 2-cell embryos were collected to detect the integration of HBV DNA in the female pronuclei of zygotes and the replication and expression of HBV genes in the 2-cell embryos using FISH, RT-PCR and immunofluoresence assay.(1) PCR detected positive bands in the tested oocyte samples fromgoldon hamster and mice. Southern blot revealed clear hybridization signals in PCR products.

研究用金黄地鼠和小鼠建立实验动物模型：将卵巢内注射HBV DNA的实验动物分成两组，一组注射后进行超排卵，收集卵巢和输卵管的卵母细胞，用PCR、Southern杂交，斑点杂交和荧光原位杂交(fluorescence in situ hybridization、FISH)检测HBV在卵母细胞内的存在和染色体上的整合；另一组超排雌鼠与正常雄鼠合笼，收集受精卵和2-细胞胚，用FISH、RT-PCR和免疫荧光检测技术分别研究HBV基因在受精卵雌原核上的整合以及在2-细胞胚中的复制与表达。

According to the clear water experiment, aeration performance of the new equipment is good with high total oxygen transfer coefficient and oxygen utilization ratio.

曝气设备的动力效率在叶轮转速为120rpm～150rpm时取得最大值，此时氧利用率和充氧能力也具有较高值。

The environmental stability of that world - including its crushing pressures and icy darkness - means that some of its most famous inhabitants have survived for eons as evolutionary throwbacks, their bodies undergoing little change.

稳定的海底环境─包括能把人压扁的压力和冰冷的黑暗─意谓海底某些最知名的栖居生物已以演化返祖的样态活了万世，形体几无变化。