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In the normal group, electron microscopy showed normal chondrocytes were oval, and cells and cell membrane were intact. In cytoplasm, there were abundant rough endoplasmic reticulum, dictyosome, and mitochondria with intact nucleus, and even chromatin. In the model group, chondrocytes exhibited obvious pyknosis and their shape was irregular. The areolae around cells disappeared; cell organs in cytoplasm coagulated to flaps which showed electron dense and was uneasy to observe; the shape of nucleus showed irregularity, and caryotin gathered densely.

电镜下正常对照组软骨细胞呈椭圆形,细胞及胞膜完整,包质内可见丰富的粗面内质网、高尔基体、线粒体,细胞核完整,染色质分布均匀;模型对照组软骨细胞明显固缩且外形不规则,细胞周晕消失,胞浆内细胞器凝成高电子密度的片状物不易分辨,细胞核不规则,染色质浓聚,散裂于核中。

The typical apoptotic morphological features appeared in MUTZ1 cells treated with 4 mmol/L VPA for 72 hours. Pyknosis of cells and nuclei, disintegration of nuclear chromatin and apoptotic body could be observed by light microscopy. Aggregation and margination of nuclear chromatin, concentration of plasm, increment of density and chromatin mass of irregular size could be observed by transmission electronmicroscope. The flow cytometric analysis indicated that the VPA could induce cell apoptosis, apoptosis rate increased in dosedependent manner, ratio of cells at G0/G1 phase increased and ratio of cells at S phase decreased in dosedependent manner, the cells were arrested at G0/G1 phase.

结果显示: VPA对MUTZ1细胞的生长抑制作用呈现时间和剂量依赖性;经4 mmol/L VPA处理MUTZ1细胞72小时后,细胞呈现典型的凋亡形态特征,光学显微镜下可见凋亡细胞胞体固缩、核固缩、核碎裂及凋亡小体;透射电子显微镜下可见凋亡细胞核染色质边集、胞浆浓缩、密度增加,胞浆内大小不规则的染色质团块;流式细胞术结果表明,细胞凋亡率随着VPA浓度的增加而逐步增高,G0/G1期细胞比例随着VPA浓度的增加而逐渐增多,S期细胞比例逐渐减低,细胞被阻滞在G0/G1期。

The typical apoptotic morphological features appeared in MUTZ-1 cells treated with 4mmol/L VPA for 72hours. Pyknosis of cells and nuclei, disintegration of nuclear chromatin and apoptotic body could be observed by light microscopy. Aggregation and margination of nuclear chromatin, concentration of plasm, increment of density and chromatin mass of irregular size could be observed by transmission electronmicroscope. The flow cytometric analysis indicated that the VPA could induce cell apoptosis, apoptosis rate increased in dose-dependent manner, ratio of cells at G0/G1 phase increased and ratio of cells at S phase decreased in dose-dependent manner, the cells were arrested at G0/G1 phase.

结果显示:VPA对MUTZ-1细胞的生长抑制作用呈现时间和剂量依赖性;经4mmol/LVPA处理MUTZ-1细胞72小时后,细胞呈现典型的凋亡形态特征,光学显微镜下可见凋亡细胞胞体固缩、核固缩、核碎裂及凋亡小体;透射电子显微镜下可见凋亡细胞核染色质边集、胞浆浓缩、密度增加,胞装内大小不规则的染色质团块;流式细胞术结果表明,细胞凋亡率随着VPA浓度的增加而逐步增高,G0/G1期细胞比例随着VPA浓度的增加而逐渐增多,S期细胞比例逐渐减低,细胞被阻滞在G0/G1期。

N perifocal tissue intracerebral hemorrhage there were rarefaction neuron,cell spaces augmentation,cell diminution,distinct demarcation of cell membrane and surrounding,and we discovered a lot of degeneration and necrosis nerve cells,cell body collapsed,pycnosis anachromasis,nucleoli disappeared. In EPO group we discovered that center area of hemorrhage shinked,nerve cells of degeneration and necrosis decreased in perifocal tissue,majority cells morphous were normal and pathological changes were light.

CH对照组在术后皮层出血中心无神经元,仅见少量胶质细胞,细胞间质空泡样改变;出血边缘区神经元稀疏,细胞间隙增大,细胞缩小,胞膜与周围分界清楚,并可见大量变性及坏死的神经细胞,表现为胞体皱缩,核固缩深染,核仁消失。rhEPO治疗组出血中心区变小,边缘区神经细胞变性坏死减少,多数存活细胞形态相对正常,病变较轻。

The construction process includes the following steps: constructing recombinant plasmid containing phhA and phhB; transforming the recombinant plasmid to hygrophilous aeromonad; and screening recombinant hygrophilous aeromonad with the recombinant plasmid.

该重组菌中含有β-酮硫解酶基因phbA和乙酰乙酰辅酶A还原酶基因phbB。其构建方法包括以下步骤:1构建含phbA和phbB基因的重组质粒;2转化该重组质粒到嗜水气单胞菌中;3筛选出含有重组质粒的重组嗜水气单胞菌。

He type of TGF β1 expression was related with lymph node metastasis and prognosis of patients. The positivity rate of TGF β1 expression only in mesenchyma in the group on lymph node metastasis was significantly lower than that in no lymph node metastasis, but the positivity rate of TGF β1 expression only in cytoplasma was higher. The prognosis of TGF β1 expression only in mesenchyma was better, but the prognosis of TGF β1 expression only in cytoplasma or both mesenchyma and cytoplasma was poor.

GF β1的阳性表达类型与肺癌的淋巴结转移及预后有关,淋巴结转移阳性组TGF β1的单一间质阳性表达率显著低于淋巴结阴性组,而单一胞浆阳性表达率高于淋巴结阴性组;半年内死亡组的TGF β1单一间质阳性表达率低于5年以上生存组,而间质胞浆均阳性的表达率及单一胞浆阳性表达率则高于5年以上生存组(P均<0.05)。

This observation provided a strong support to the point that most leaves export the most of assimilates in the light time. Plasmodesmal densities between SE/CC, CC/PP, PP/PP and PP/BSC (bundle-sheath cell) decreased in weak light. Plasmodesmata were observed between CC/SE (nacreous-walled sieve element), PP/BSC in branch veins in normal light intensity, but not in weak light. Thus apoplasmic pathway may be the main mode of transport of assimilates in weak light, however symplasmic pathway may be the main mode of transport of assimilates in normal light intensity.

在筛管/伴胞、伴胞/韧皮薄壁细胞、韧皮薄壁细胞/韧皮薄壁细胞和韧皮薄壁细胞/维管束鞘细胞之间的胞间连丝密度都在弱光条件下下降,在正常光照强度下支脉筛管/伴胞和韧皮薄壁细胞/维管束鞘细胞之间可以观察到胞间连丝,而在弱光下几乎观察不到胞间连丝的存在,所以同化物的运输在弱光条件下可能以质外体运输为主,而在正常光照强度下,共质体运输可能是主要的运输方式。

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

Sour orange and sweet orange may be the hybrid of pummelo and mandarin.

6根据胞质基因组的差异和聚类分析的结果对一些生物型的起源进行了分析,得出如下一些结论:葡萄柚是柚与甜橙的杂种,可能是与柚的回交种;本研究所包括的两个来檬生物型,其中之一的起源与枸橼、柚和澳洲指橘有关,另一个被误认为是枸橼的来檬生物型最后被认定为墨西哥来檬,来源与大翼橙和枸橼有关;粗柠檬是枸橼与宽皮橘的杂种;红黎檬可能是宽皮橘与粗柠檬的杂种;酸橙和甜橙为柚与宽皮橘的后代。

The morphology and structure of reconstructed tissue was detected by microscope and scanning electron microscope.Results:(1) Compared with the control group, the cellular proportion of laminin group increased in 62 ~M phase, and decreased in Go~Gi phase significantly. As shown by the microscope, the cells of control group were in low density. The cells in mass connected tightly. The microfilament appeared reticular formation. The nucleus were the same in size. The cells of laminin group were in high density and put out so many lamellipodia, filopodia, which connected with the surrounding cells. The microfilament increased, elongated, and changed from reticulodromous to sarciniform, which reached to the pseudopods. The nucleus were different in size .(2) As shown by the inverted microscope and the cell growth curve, comparing with the controlgroup, cells of each test group increased evidently. The cellular proportion of each test group increased in S phase and G2 ~M phase, and decreased in Go~Gi phase significantly, but there was no considerable interations between LN and EGF;(3) As shown by the morphological observations, the cultured cat corneal endothelial cells formed an integrated membrane, and attached to the Descemets membrane closely, which was similar to the natural tissue. The cells connected tightly to each other, and some of them arranged in hexagon approximately.

结果:(1)层粘连蛋白组处于G_2~M期细胞比例较对照组显著提高,Go~G_1期细胞比例显著下降,提示层粘连蛋白促进内皮细胞DNA合成,及细胞分裂增殖;光镜下,对照组细胞分布成团状,细胞密度较低,细胞间连接紧密,细胞内微丝结成网状,细胞核大小一致;与对照组相比,层粘连蛋白组细胞生长旺盛,细胞密度高,向周边伸出大量板状及丝状伪足,细胞内微丝增多、拉长、集结成束,伸入伪足中,细胞核形状大小不一致;(2)倒置显微镜观察及细胞生长曲线显示,各组细胞数目随时间增加而明显增多,各实验组较对照组增生显著,EGF和LN联合应用组各时间点细胞数目最高;实验组处于S期和G_2~M期细胞数目增加,Go~G_1期细胞数目减少;提示EGF、LN单独及联合应用均可促进细胞增殖,但尚不能认为二者有交互作用;(3)倒置显微镜下,组织培养的猫角膜内皮细胞排列成密集的单层,细胞间连接紧密;组织学观察发现,培养的猫角膜内皮细胞形成完整的内皮层,贴附于脱水基质的后弹力膜上,与正常的角膜内皮组织结构相似;扫描电镜下,组织培养的猫角膜内皮细胞间紧密镶嵌排列,可见某些细胞呈近似六边形排列,细胞大小不甚一致,胞核清晰。

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