细胞

The results showed that the lesion of X-rays oncells displayed gradually attenuation with depth (or cellsurvival fraction displayed gradually increase with depth)while the lesion of heavy ions on cells followed Bragg curve(or cell survival fraction was inverse Bragg cueve).(2)Thedynamic change of micronuclei and cell survival inhepatoma SMMC-7721 irradiated by 25MeV/u〓 wasstudied.

采用HeLa、B16两种细胞分别研究了X-射线和重离子在水介质中入射的深度与相应细胞的存活率（1-失活率），结果表明：X-射线对细胞的损伤随深度而逐渐衰减（或细胞存活随深度逐渐增加），而重离子对细胞的损伤则为Bragg曲线（或细胞存活为倒Bragg曲线）。

The chondrocytes of different generation were observed with light-microscope and transmission electron microscope for cellular growth and ultromicrostructure, with the method of MTT assay for grow curve and proliferation, with alcian blue test for GAG of ECM, with immuocytochemistry and RT-PCR (reverse transcript -polymerase chain reaction)for type Ⅱcollagen, with flow cytometry for cell life cycle ,histochemistry for S-A-β-galand so on by which to identify cataplasia and senescence of chondrocytes cultured in vitro.

对软骨细胞退变老化进行观察，采用相差显微镜观察其生长情况，透射电镜观察细胞结构，阿力新蓝染色检测胞外基质硫酸GAG含量和结构，免疫细胞化学法和RT-PCR方法检测Ⅱ型胶原鉴定软骨细胞，以MTT比色法描绘生长曲线、检测生长状态，组化法检测老化相关β-半乳糖苷酶，流式细胞仪分析细胞周期和增殖指数。3。

The third passage chondrocytes were divided into blank group, different desity PAP groups, different desity glucosaminsalfate groups which were passaged to 4th generation and contrast to the 2nd passage group. The chondrocytes of different groups were detected with the method of histochemistry for S-A-β-gal,and with alcian blue test for the content and constructure of GAG of ECM, immuocytochemistry for type Ⅱcollagen and PCNA, MTT assay for proliferation, RT-PCR for type Ⅱcollagen and Aggrecan, flow cytometry for cell life cycle and proliferation index,by which to observe PAP's function regarding to the appearance and functional status in the process of chondrocyte's cataplasia and senescence.

将P3软骨细胞分为空白对照组、鹿茸多肽不同浓度组、硫酸氨基葡萄糖不同浓度组进行传代培养，同时以P2代软骨细胞为对照组，进行组化检测老化相关β-半乳糖苷酶，阿力新蓝染色检测胞外基质硫酸GAG含量和结构，MTT比色检测增殖，免疫细胞化检测PCNA和Ⅱ型胶原，RT-PCR检测Ⅱ型胶原、Aggrecan蛋白，流式细胞仪分析细胞周期和增殖指数等方法，对鹿茸多肽抗软骨细胞退变老化进行分子生物学研究。4。

The 3rdpassage chondrocytes were divided into blank group, different concentration PAP groups,different concentration glucosaminsalfate groups and were sequently passaged to 4thgeneration. The 2nd passage chondrocytes was contrasted as young cells group. Thechondrocytes of different groups were detected with the methods of histochemistry forS-A-β-gal, and with alcian blue test for the content and constructure of GAG of ECM,immuocytochemistry for typeⅡcollagen and PCNA, MTT assay for proliferation, RT-PCRfor typeⅡcollagen and Aggrecan, flow cytometry for cell life cycle and proliferationindex,by which to observe PAP"s function regarding to the appearance and functional status inthe process of chondrocyte"s cataplasia and senescence.

将P3软骨细胞分为空白对照组、鹿茸多肽不同浓度组、硫酸氨基葡萄糖不同浓度组进行传代培养，同时以P2代软骨细胞为对照组，进行组化检测老化相关β-半乳糖苷酶，阿力新蓝染色检测胞外基质硫酸GAG含量和结构，MTT比色检测增殖，免疫细胞化检测PCNA和Ⅱ型胶原，RT-PCR检测Ⅱ型胶原、Aggrecan蛋白，流式细胞仪分析细胞周期和增殖指数等方法，对鹿茸多肽抗软骨细胞退变老化进行分子生物学研究。4。

4DRG was co-culture with sciatic nerve segment in 10鸖 DMEM;the axons were longer and surround the sciatic nerve segment which was regard as anew evidence for chemotropism.

结果：(1)在无血清条件下单独培养的DRG，背根神经节的轴突数目众多，外形纤细弯曲，不成束，并且施万细胞和成纤维细胞稀少，所以可以排除两者对轴突生长的影响，为观察来源于变性坐骨神经段的可溶性因子对轴突生长的作用提供了有利条件；(2)在无血清条件下DRG和变性坐骨神经段联合培养，①先单独培养DRG，4天后待神经元轴突长出，再与坐骨神经段联合培养，观察到神经元的轴突数目减少，外形挺直，部分轴突之间相互粘附成束；②变性坐骨神经段和DRG同时联合培养，神经元的轴突数目明显减少，外形粗壮，轴突之间相互粘附成束；(3)有血清条件下单独培养DRG，轴突数目较多，外形挺直，长短不一，部分神经元的轴突之间相互粘附，施万细胞和成纤维细胞数目众多，观察到的轴突生长情况受到施万细胞和成纤维细胞的直接或者间接的影响。

In the brain of adult rat, the positive immunohistochemical product of lSL-l (ISL-l-positive) was mainly located in the neuronal nucleus and found in discrete regions except to brain cortex, such as the Purkinje cell layer and the granular cell layer of cerebellum, the granular cell layer and the pyramidal cell layer of hippocampus, the mitral cell layer, the internal and external plexiform layer, the granular cell layer and the granular cell layer of olfactory bulb and so on, and several nuclei of the hypothalamus, midbrain and pons, such as claustrum, anterior olfactory nucleus, accumbens nucleus, caudate-ptamen, pallidum, substantia nigra, striatum, islands of Callaje, mammillary nucleus, anterior pretactal nucleus, habenular nucleus, amygdaloid nucleus, cuneate nucleus, rubral nucleus, gigantocellular reticular nucleus and so on.

在正常成年大鼠脑中，同源框基因islet-1表达产物（ISL-1）免疫组织化学阳性物质广泛分布于除大脑皮层外的神经细胞的细胞核内，ISL-1阳性神经元密集分布于小脑Purkinje细胞层和颗粒细胞层、海马的颗粒细胞层和锥体细胞层、嗅球的内丛层、外丛层、颗粒细胞层及僧帽细胞层等，另外在丘脑、中脑和桥脑的一些重要神经元核团均有分布，如，屏状核、前嗅核、伏核、尾壳核、苍白球、黑质、纹状体、Calleja岛、乳头体核、前顶盖前核、缰核、杏仁核、楔束核、红核网状巨细胞核等。

Tetraploid embryos could be produced by electrofusion at the stage of two-cell embryos, which could develop to blastocysts followed by fusion of cytoplasm and nucleus and cleavage in vitro. During the fusion of cytoplasm, the DNA methylation levels of the fused embryos are as high as these of two-cell diploid embryos in vivo Then the embryos are rapidly demethylated when the nucleus begin to fuse, resulting in the lowest DNA methylation levels when the nucleus are fused completely. After that, the DNA methylation levels of the fused embryos are gradually increased until the morula stage. However, whereas an asymmetric distribution of DNA methylation is established in vivo-derived blastocysts with a higher methylation level in the inner cell mass than that in the trophectoderm, we can not detect the asymmetric distribution in most in vitro-derived tetraploid blastocysts.

结果表明：利用电融合方法制备的小鼠四倍体胚胎在体外培养体系中经历细胞质融合、细胞核融合及细胞继续分裂发育直到囊胚期的过程，在细胞质融合的时候胚胎卵裂球同体内体外培养二倍体胚胎一样，呈现高度甲基化状态；在细胞核开始融合的时候，甲基化水平急速下降，在细胞核完全融合的时候甲基化水平达到最低点；随着胚胎继续分裂，胚胎甲基化水平逐渐增加，在桑葚胚期甲基化水平最高；但是囊胚期四倍体胚胎内细胞团同滋养层细胞甲基化荧光信号没有差别，这与体内体外培养二倍体囊胚内细胞团细胞甲基化荧光强度高于滋养层细胞甲基化荧光强度不同。

Tetraploid embryos could be produced by electrofusion at the stage of two-cell embryos, which could develop to blastocysts followed by fusion of cyto-plasm and nucleus and cleavage in vitro. During the fusion of cytoplasm, the DNA methylation levels of the fused embryos are as high as these of two-cell diploid embryos in vivo Then the embryos are rapidly demethylated when the nucleus begin to fuse, resulting in the lowest DNA methylation levels when the nucleus are fused completely. After that, the DNA methy-lation levels of the fused embryos are gradually increased until the morula stage. However, whereas an asymmetric distribu-tion of DNA methylation is established in vivo-derived blastocysts with a higher methylation level in the inner cell mass than that in the trophectoderm, we can not detect the asymmetric distribution in most in vitro-derived tetraploid blastocysts.

结果表明：利用电融合方法制备的小鼠四倍体胚胎在体外培养体系中经历细胞质融合、细胞核融合及细胞继续分裂发育直到囊胚期的过程，在细胞质融合的时候胚胎卵裂球同体内体外培养二倍体胚胎一样，呈现高度甲基化状态；在细胞核开始融合的时候，甲基化水平急速下降，在细胞核完全融合的时候甲基化水平达到最低点；随着胚胎继续分裂，胚胎甲基化水平逐渐增加，在桑葚胚期甲基化水平最高；但是囊胚期四倍体胚胎内细胞团同滋养层细胞甲基化荧光信号没有差别，这与体内体外培养二倍体囊胚内细胞团细胞甲基化荧光强度高于滋养层细胞甲基化荧光强度不同。

Tetraploid embryos could be produced by electrofusion at the stage of two-cell embryos, which could develop to blastcysts fellowed by fusion of cytoplasm and nucleus and cleavage in vitro.After fusion of cytoplasm, the DNA methylation levels of the fused embryos was very high as well as two-cell diploid embryos in vivo.Then the embryos was rapiddly demethylated when the nucleus begin to fuse, resulting the lowest DNA methylation levels when the nucleus fused completely.After that, the DNA methylation levels of fused embryos were gradually increased until the blastocysts stage.However, whereas an asymmetric distribution of DNA methylation was established in an vivo-derived blastocysts with a higher methylation level in the inner cell mass than in the trophectoderm, in most vitro-derived tetraploid blastocysts, we can not detect the asymmetric distribution.

结果表明：利用电融合方法制备的小鼠四倍体胚胎在体外培养体系中经历细胞质融合、细胞核融合及细胞继续分裂发育直到囊胚期的过程，在细胞质融合的时候胚胎卵裂球同体内体外培养二倍体胚胎一样，呈现高度甲基化状态；在细胞核开始融合的时候，甲基化水平急速下降，在细胞核完全融合的时候甲基化水平达到最低点；随着胚胎继续分裂，胚胎甲基化水平逐渐增加，在囊胚期甲基化水平最高；但是囊胚期四倍体胚胎内细胞团同滋养层细胞甲基化荧光信号没有差别，这与体内体外培养二倍体囊胚内细胞团细胞甲基化荧光强度高于滋养层细胞甲基化荧光强度不同。

RESULTS:①Cytocompatibility of PEG-PLA-PGL membranes with HUVECs: The observation result of phase contrast microscopy showed that, endothelial cells planted on the PEG-PLA-PGL membranes began to attach and stretch after being planted 4-6 hours. Three days later, cells grew in colonies rapidly, after 5 days, colonies began to fuse and seemed like cobble-stone. The cells were shuttle or polygon in shape after passages.

结果：①人脐静脉内皮细胞与聚乙二醇-聚乳酸-聚谷氨酸三嵌段共聚物膜片的细胞相容性：相差显微镜下种植在聚乙二醇-聚乳酸-聚谷氨酸三嵌段共聚物膜片上的细胞4~6 h后即开始贴壁、伸展，3 d后细胞开始呈集落样生长，生长较快，5 d后细胞集落开始融合，呈现特征性的鹅卵石样形态，经多次传代后，细胞呈长梭形或多角形，与对照组相比无明显差异。

This one mode pays close attention to network credence foundation of the businessman very much.

这一模式非常关注商人的网络信用基础。

Cell morphology of bacterial ghost of Pasteurella multocida was observed by scanning electron microscopy and inactivation ratio was estimated by CFU analysi.

扫描电镜观察多杀性巴氏杆菌细菌幽灵和菌落形成单位评价遗传灭活率。