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Compared with the corresponding BEAS-2B cells, the shapes of cells and nuclears, the shapes and quantity of cell organs were not significantly changed in the 1st, 2nd, 5th passage BEAS-2BNNK cells. The 10th, 15th passage BEAS-2BNNK cells, which had swelled cell and cell organs and abnormal nuclear and enlarged the nucleoli gradually and increased the quantity of the cell organs and activated their function, showed the characteristic of transformation cells. The 20th, 25th passage BEAS- 2BNNKcells, which had obvious aberration of the cell and nucleoli and had cataclasm of nucleoli and decreased cell organs, showed the characteristic of tumor cells.

与BEAS-2B相比较BEAS-2BNNK第1、2、5代细胞及胞核形态、细胞器形态及数量无明显差别;第10、15代细胞出现肿胀,细胞核逐渐变形,出现核畸形,核仁明显增大,边集,细胞器肿胀,数量明显增多,功能活跃具明显的转化细胞特征;第20、25代细胞及胞核明显畸变,出现明显的核碎裂及多个核仁,细胞器明显减少,具明显的肿瘤细胞特征。

More cleavage phases can be counted by FISH than G-band dyeing on the diagnosis of mosaicism kyrotype, even many intermediate stage cells can be analyzed by FISH procedure. Time was saved and more precise results can be found.

与常规染色体核型分析方法比较,FISH技术检测嵌合体核型能够观察到更多的分裂相,还可分析核膜完整染色体未分散的间期细胞,计数更多的细胞,为临床提供更准确的数据,且省时。

The porine is the polyembryony animal, many obtains the ovocyte from the porine ovary although but the maturity quite to be low; Because the ROSI technology is does not have to grow the mature sole round sperm cell to pour into directly completely in the ovicell nature, jumped over the spermatozoon in to pass through the physiology and the biochemistry, if the ovicell nature mature or the activation degree were insufficient, added the round immature sperm cell maturity quite inferior reason, very possibly Causes the ROSI micro fertilization defeat.

猪是多胎动物,从猪卵巢上获得的卵母细胞数量虽然较多,但成熟度比较低;由于ROSI技术是将没有完全发育成熟的单一圆形精细胞直接注入卵胞质内,跳越了精子在穿过透明带和卵质膜等过程中所发生的生理和生化反应;如果卵胞质成熟或活化程度不够,加之圆形未成熟精细胞成熟度比较差等原因,很可能造成ROSI受精的卵母细胞内部激活因子蓄积减少和生成不足,引起精核解聚困难、精圆核不能形成、第二极体不能排出、卵母细胞孤雌发育率增加等,使ROSI显微受精失败。

ResultsMorphological features with diagnostic value were established as follows:①Irregular or round-mesh basket type nucleolus;②Malformed nucleus and lobate nucleus with fine bridge;③Perinuclear microfilaments;④Nuclear bodies;⑤Nuclear inclusion bodies;⑥Marked cell pleomorphism arranged irregularly,like a disordered stone pavement;⑦Strange cell and/or giant cancer cell were found;⑧Peculiar papillae or cell clusters were formed;⑨Specific function structure on the surface of cell differentiated poorly.

结果 确定了具有诊断意义的形态学特征:①不规则奇形核仁或网孔状核仁;②畸形核,分叶核伴细桥-核间桥形成;③核旁微丝;④核体;⑤核包涵体;⑥细胞重度多形性,排列紊乱,似乱石铺堆;⑦出现奇形细胞和/或癌巨细胞;⑧形成奇形乳头和奇形细胞簇;⑨细胞表面特异功能结构分化不良。

Grew on the normal temperature, it was shown that the deposits of calcium antimonate being the indicator for Ca(superscript 2+) localization mainly concentrated within the vacuoles and intercellular spaces and there was also some Ca(superscript 2+) deposits in cell walls. But when Garyota urens L. was treated by the temperature of 2℃ for 48 h, the level of Ca(superscript 2+) increased in cytoplasm and plasma membrane, but decreased in vacuoles and intercellular spaces considerably. At the same time, the ultrastructure of chloroplasts suffered from chilling: the membrane of chloroplasts had been damaged, the layer of thylakoids was exiguous and unclear, the photosynthetic rate decreased evidently. And when Garyota urens L. was treated by the temperature of 2℃ for 120 h, the deposits of Ca(superscript 2+) mainly concentrated within the cytoplasm, nucleus and plasma membrane and there was also some Ca(superscript 2+) deposits in vacuoles, and the ultrastructure of some cells was simultaneously damaged severely: Chloroplasts structure, vacuole membrane and nuclear membrane had been damaged fully, the structure within the cell had become unclear, and the cell only have respiration.

研究结果表明,未经低温处理的董棕幼苗叶肉细胞,焦锑酸钙沉淀颗粒大量出现在液泡和细胞间隙中,细胞壁中也可见少量沉淀,而细胞基质中则看不到焦锑酸钙沉淀;经2℃ 48 h低温处理后,细胞基质和细胞膜上焦锑酸钙沉淀增加,而液泡和细胞间隙中的焦锑酸钙沉淀则显著减少,并且超微结构已初步显示出寒害的特徵,叶绿体外膜部分破损,类囊体片层稀疏且排列不规则,光合速率明显下降等;经2℃ 120 h低温处理后,细胞间隙内的焦锑酸钙沉淀极少,有的也紧贴在细胞外壁上,而细胞基质和细胞膜上则分布有非常多的焦锑酸钙沉淀,在核基质和液泡中也可见到少量的焦锑酸钙沉淀,并且超微结构遭到了显著破坏,叶绿体结构完全被破坏,核膜与液泡膜严重破损,内部结构模糊,细胞只表现为呼吸作用,不进行光合作用。

Glioma is still one of refractory disease in the neurosurgical field; the development of new primary and adjuvant treatment is vital. Recently, the gene therapy of glioma is developed rapidly and there are many methods about the gene therapy that include: suicide gene therapy, immunologic gene therapy, drug resistangce gene therapy, angiostatin gene therapy and so on. The sucide gene therapy is the most potential approach of antitumer, these nonmammalian genes encode enzyme that convert nontoxic prodrugs into highly toxic metablites. Cells transfected with suicide genes are targeted for specific negative selection, witch can be induced by administrtion of the corresponding produg. Among the enzyme/produg combinations, two of the best characterized system are herpes simplex virus thymidine kinase /ganciclovir and Escherichia coli cytosine deaminase /5-flourocytosine (5-FC). The formor can convert the antiviral nucleoside analogs acyclovir , ganciclovir to their nucleoside monophosphate derivatives, the monophosphate forms are subsequently phosphorylated by endogenus cellular kinases to triphosphates, these molecules are potent inhibitors of DNA synthesis.

近年来脑胶质瘤的基因治疗发展迅速,应运而生的方法有自杀基因、免疫基因、多药耐药基因以及抗血管生成基因等,其中自杀基因被认为是最有前景的基因治疗方法,它又称病毒介导的酶/药物前体疗法,是利用转基因技术将哺乳动物细胞中所不含有的自杀基因转入到哺乳动物肿瘤细胞中,该基因表达的产物可将无毒的药物前体转化为毒性药物,从而选择性杀伤该肿瘤细胞,常用的自杀基因有单纯疱疹病毒-胸苷激酶基因和大肠杆菌胞嘧啶脱氨酶基因,前者催化无毒性抗病毒核苷类似物如丙氧鸟苷、无环鸟苷等成为单磷酸核苷衍生物,然后在内源性细胞激酶作用下转化为具有明显毒性的三磷酸核苷,作为DNA合成链的终止剂和DNA合成酶的抑制剂,干扰细胞DNA的合成;后者编码的胞嘧啶脱氨酶可催化5-氟胞嘧啶(5-FC)脱氨成为5-氟尿嘧啶(5-FU),然后代谢为有毒性的5-氟尿嘧啶-5′三磷酸(5-FUTP)和5-氟-2′脱氧尿嘧啶-5′磷酸(5-FdUTP),5-FUTP通过与UTP竞争性结合而抑制mRNA和tRNA的合成,5-FdUTP则作用于胸苷合成酶,导致TMP衰竭而阻止DNA的合成,最终诱导肿瘤细胞凋亡。

Results: Of all the semen samples, 8 were HCMV positive, 4 HSV-Ⅱ positive, but none were both HCMV and HSV-Ⅱ positive. HCMV late antigens were positively and HCMV early antigens negatively expressed in the spermatogenic cells of the 8 HCMV positive cases. In the 4 HSV-Ⅱ positive cases, 3 were positively and 1 weakly positively expressed. In the semen of the 12 positive cases were found large numbers of immature spermatogenic cells, with different manifestations of apoptosis, such as chromatin pycnosis, vacuoles, damaged nuclear membrane, and apoptotic bodies, but without virus infection-induced specific morphological alteration.

结果:83例精液样本PCR检测HCMV阳性8例,HSV-Ⅱ阳性4例,未发现2种病毒同时阳性病例。8例HCMV阳性病例ICC标记生精细胞HCMV晚期抗原均见阳性表达,HCMV早期抗原均为阴性表达;4例HSV-Ⅱ阳性病例ICC标记生精细胞3例阳性表达,1例弱阳性。12例病毒阳性病例均发现较多未成熟生精细胞,并有不同程度的核染色质固缩、出现空泡、核膜破损、核崩解、凋亡小体等凋亡现象,但未能找到病毒感染的特异性形态学改变。

Fig.1 SHEE cultured on coverslide, the living cells were growing in single layer with rich cytoplasm, the nuclei were uniform in size with a nucleolus ph ×400 Fig.2 SHEE had a nucleus with ellipse shape, large nucleolus and the cytoplasm contained mitochondria and tonofibrilEM ×10 000 Fig.3 SHEE was spherical in shape, with pseudopods attached on petri dish and abundant villi on cell surface SEM ×5 000 Fig.4 Same as in Fig.3, cell attached on petri dish, appeared stellate or polygonal in shape, with abundant pseudopods and cytoplasmic processes. Protrusive nuclear region in central part of the cell had more micro-villi SEM ×5 000 Fig.5 Chromosomes of SHEE cells belonged to diploidy type Giemsa ×1 000 Fig.6 The SHEE cells of stained in dark brown by Ki67 immunohistochemistry were the proliferative cells Immunohistochemistry ×400 Fig.7 In SHEE cell culture, the nucleus stained red or pink by PI was dead cell, the green nucleus was living cell Fluorescent ×400 Fig.8 The cell labeled by TdT was apoptotic cell in which the chromatin of nucleus condensed in block, a pyknotic nucleus in the upper right conner was seen TdT labeled ×400

图1 SHEE培养在盖坡片上,活细胞单层生长,胞浆较丰富,细胞核大小一致,有核仁×400 图2 SHEE培养细胞细胞核椭圆形,核仁较大,胞浆有较丰富的线粒体和张力原纤维EM ×10 000 图3 SHEE细胞呈球状,有伪足贴壁,表面有密集微绒毛SEM ×5 000 图4 同上细胞贴壁,呈星状或多角形,有丰富伪足和胞浆突,核区隆起有较多微绒毛SEM ×5 000 图5 SHEE细胞染色体仍属二倍体Giemsa染色×1 000 图6 SHEE细胞Ki67免疫组织化学染棕黄色为增殖细胞×400 图7 SHEE培养细胞出现死细胞,胞核和胞浆PI染色呈红色或淡红色,蓝色细胞核为活细胞荧光显微镜×400 图8 细胞TdT标记阳性为凋亡细胞,染色质凝集呈块状,右上角有一固缩细胞核TdT标记×400

Pantaleo G,Graziosic,Demarest JF,et al.HIV infection is active and progressive in lymphoid tissue during the clinically latent stage of disease[J].Nature,1993,362:355~360[5]Moss PAH,Rowland-Jones S,Frodsham PM,et al.Persistent high frequency of human immunodeficiency virus-specific cytolytic T cell in peripheral blood of infected donors[J].Proc Natl Acad Sci USA,1995,92:5773~5777

一些研究显示在无免疫肌肉大夫压力下体外传代HIV的状况类似体内免疫肌肉系统全面崩溃的感染血液末期的状况:HIV变异株相对少,株间变异小,但毒力强、复制快、病毒痔疮产量高,可形成多核巨细胞疑难等等;HIV在适应了体外培养的环境和宿主细胞疑难、失去了选择大夫压力后,少数变异株在传代过程中经逐步稀释后逃脱优势株的抑制逐渐被固定下来而成为新的优势株[7]。

Pantaleo G,Graziosic,Demarest JF,et al.HIV infection is active and progressive in lymphoid tissue during the clinically latent stage of disease[J].Nature,1993,362:355~360[5]Moss PAH,Rowland-Jones S,Frodsham PM,et al.Persistent high frequency of human immunodeficiency virus-specific cytolytic T cell in peripheral blood of infected donors[J].Proc Natl Acad Sci USA,1995,92:5773~5777

一些研究显示在无免疫压力下体外传代HIV的状况类似体内免疫系统全面崩溃的感染末期的状况:HIV变异株相对少,株间变异小,但毒力强、复制快、病毒产量高,可形成多核巨细胞等等;HIV在适应了体外培养的环境和宿主细胞、失去了选择压力后,少数变异株在传代过程中经逐步稀释后逃脱优势株的抑制逐渐被固定下来而成为新的优势株[7]。

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Finally, according to market conditions and market products this article paper analyzes the trends in the development of camera technology, and designs a color night vision camera.

最后根据市场情况和市面上产品的情况分析了摄像机技术的发展趋势,并设计了一款彩色夜视摄像机。

Only person height weeds and the fierce looks stone idles were there.

只有半人深的荒草和龇牙咧嘴的神像。

This dramatic range, steeper than the Himalayas, is the upturned rim of the eastern edge of Tibet, a plateau that has risen to 5 km in response to the slow but un stoppable collision of India with Asia that began about 55 million years ago and which continues unabated today.

这一引人注目的地域范围,比喜马拉雅山更加陡峭,是处于西藏东部边缘的朝上翻的边框地带。响应启始于约5500万年前的、缓慢的但却不可阻挡的印度与亚洲地壳板块碰撞,高原已上升至五千米,这种碰撞持续至今,毫无衰退。