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细胞性

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While the tissue spaces surrounding a few blood vessels wasAl and Fg positive,no Al or Fg positive cells were observed.In antemortem injurygroup,diffuse subarachnoid hemorrhage,cerebral edema,swelling or pyknotic neu-rons could be observed.The axons showed irregular swelling and disconnection at1~3h,marked swelling and disconnection at 6h,and retraction ball at 15h whichwas more remarkable at 24h after injury.The space between myelin sheaths andaxons was increased at 3~6h after injury.Tortuous and wavelike myelin sheathswhich adhered on axons incompletely,or even peeled off could be found from 15hto 24h after injury.Perinuclear lysis of Nissl bodies began at 24h after injury.Thenumber of GFAP positive cells in cerebrum and brain-stem increased significantlyfollowed by decrease,and then increased again,but the time courses of the changesin different areas of brain were not same.Al and Fg positive neural cells,mainlysurrounded blood vessels,with diffuse or peripherally distributed positive matter incytoplasm could be observed at 0.5h after injury.The number of Al or Fg positivecells and the intensity of immunoreaction increased with the time of injury.The areaof SYN positivity in medulla oblongata and pons decreased notably 3~6h afterinjury,then return to normal levels and continued to 24h after injury.

生前损伤组,可见广泛蛛网膜下腔出血,脑组织水肿,神经细胞肿胀,晚期神经元固缩;伤后1~3h见部分神经轴突不规则增粗、断裂,伤后6h断端膨大,伤后15h可见收缩球,至伤后24h更为明显;伤后3~6h可见部分神经髓鞘与轴突之间的间隙增宽,伤后15h髓鞘明显曲折,不完全附着在轴突两侧,甚至剥脱,持续到伤后24h;核周尼氏体减少在伤后24h才开始出现;同一部位的GFAP阳性细胞数目随损伤时间发生改变,先增多(最早在伤后0.5h),达到高峰后减少,其后又有增多趋势,但不同部位的GFAP阳性细胞数目增减的时间过程不尽相同,同时,大脑中的GFAP阳性细胞数目也有改变;伤后0.5h,可在脑干组织中见到Al和Fg阳性神经细胞,主要位于血管周围,阳性物在胞浆中呈弥散性分布,但部分细胞的阳性物仅分布于靠近胞膜的胞浆中而呈环状,随损伤时间延长,阳性细胞数目增多,反应强度增加;伤后3~6h,延髓及桥脑中的SYN阳性物面积减少,其后恢复到正常水平,并持续到伤后24h。

①Rat MSC and VSMC were cultured and identified, respectively. MSC were labeled with DAPI firstly, and then co-cultivated with VSMC. The changes of morphology and ultrastructure of co-cultured cells were observed. Immunfluorescence analysis was performed by using monoclonal antibodies against specific antigen.②We established the regulatable system in two steps: a stable MSC line expressing rtTA has been constructed and characterized firstly by transfected with pUHD 17-1hyg and then selected by hygromycin B; in a second step, this line was used for trandfer the AT2R gene to MSC to get the well establishing double stable MSC lines;③The expression of AT2R regulated by doxycycline was evaluated by western blot;④The MSCs were transduced into rat carotid arteries with regulatable AT2R gene after the establishment of rat carotid balloon injury restenosis model. The intimal/medial area ratio were measured by digital analysis system.

研究方法:(1)密度梯度离心法及胶原酶消化法分别培养原代大鼠MSC及VSMC,细胞共培养并行免疫荧光化学染色和透射电镜观察超微结构;(2)组成受Dox调控的哺乳动物表达系统的四种成分的转化、扩增及提纯并酶切鉴定;(3)采用常规分子生物学方法连续两个回合转染体外培养的MSC,并分别采用发光计检测不同细胞克隆萤光素酶活性改变以及RT-PCR方法检测AT2R目的基因mRNA表达情况,根据各个细胞克隆受Dox调控表达的程度,选择低背景、高诱导表达AT2R的细胞系,作为双重稳定MSC细胞系;蛋白免疫印迹法观察该细胞系在Dox调控下AT2R表达的时相性、持续性及在不同浓度Dox调控下的表达情况;(4)建立大鼠颈动脉球囊损伤动物模型,将双重稳定MSC在术中导入血管,分别于14 d、28 d进行病理切片,检测可调控AT2R对新生内膜增生的影响;采用RT-PCR免疫组织化学免疫荧光等技术观察AT2R基因在新生内膜中的表达以及细胞外基质成分表达的改变,TUNEL法检测血管组织中细胞凋亡的变化情况。

On day40 after infection,donor derived Thy-1.1~+ CD8Tm cells were detectable in various organs including peripheral blood,spleen,lymph nodes,and liver.These cells were CD44~,CD62L~/CD62L~ in phenotype and more importantly,these cells were readily detectable for intracellular IFN-γsecretion several hours after ex vivo restimulation with OVA_(257-264) peptide.

感染40天后,可以在包括外周血、脾脏、淋巴结和肝脏在内的脏器中检测到一定比例的Thy-1.1~+ CD8 T细胞,这群细胞具有记忆性CD8T细胞的表型特征,即CD44~,CD62L~/CD62L~,更重要的是,这群细胞在体外OVA_(257-264)肽再刺激数小时后即可检测到细胞内IFN-γ分泌。

Finally, we used GF109203X, Rottlerin, PKC-delta antisense RNA and PKC-epsilon siRNA to inhibit PKC activation or expression, and then observed their effects on TRAIL-induced apoptosis.

当以Rottlerin处理细胞时,均会增加细胞对於TRAIL的感受性。除了H1299细胞,p53能透过藉由Rottlerin抑制PKC-delta的活性,进而增加细胞对TRAIL诱导细胞凋亡的情形。

Under normal temperature, anthers of not fertility restoration stopped developing in the stage of archesporical cell; 1-2 comers of a few anther could differentiate and form archesporical cell, sporogenous cells and microspore mother cell, but microspore mother cell disintegrated on the first meiotic division.

常温下,未发生育性恢复的花药在孢原细胞期就停止发育;少数花药1~2个角隅处可分化形成孢原细胞,造孢细胞,花粉母细胞,但花粉母细胞在减数分裂的第一次分裂时解体。

Methods:Human CD80 cDNA was transfected into B16 melanoma cells by lipofectin-mediated gene transfer before the expansion of tumor cells were tested by MTT method in vitro;C57BL/6 mice were inculated subcutaneously either mock-transfected B16 cells (B16-neo)or CD80-transfected B16 cells (B16-CD80),then the latency,survival times and tumor mass growth were investigated.The lymphocytes were examined for both proliferation indices and specific cytotoxic activity by MTT method and improved LDH-releasing method,after syngenic Mixed Lymphocyte tumor cultures.

通过脂质体介导将CD80基因导入小鼠黑色素瘤B16细胞后,利用SABC法检测CD80的表达;MTT法测定肿瘤细胞体外增殖能力;将肿瘤细胞接种至同源小鼠皮下后观察成瘤期、荷瘤小鼠存活期及肿瘤结节生长速度;在同源淋巴细胞肿瘤细胞混合培养后,通过测定淋巴细胞增殖指数和CTL杀伤活性检测肿瘤细胞的免疫原性。

Results IL-6R immunoreactive cells were observed in the anterior of optical area,the ventromedial nucleus,the periventricular nucleus and arcurate nucleus of the hypothalamus.The strongly stained cells are concentrated in the granule cell layer of the dentdate gyrus and the pyramidal cells of the CA1-CA2 fields of the hippocampus.IL-6R positive cells are also detected in the cortex, the ventromedial nucleus of the thalamus,bed nu stria,olfaltory tubercle.

结果 IL-6R 免疫反应性细胞主要分布在下丘脑的视前区、腹内侧核、室旁核和弓状核;在海马, IL-6R 阳性细胞呈强阳性,密集分布于齿状回颗粒细胞层和 CA1~ CA2锥体细胞层;在大脑皮层、嗅结节、丘脑腹内侧核、终纹床核等处也有 IL-6R 阳性细胞。

As exposed to optics microscope and inverted microscope,compared to control,SW480 cells with Giemsa staining that treated with the H2RA(cimetidine、nizatidine) took on malignance declining as crimpled firstly,cellular bulk and heteromorphism dimimished,the amount of rounded cell manifolded,nucleolus bulk lessened,dyeing thin,the amount of nucleolus decreased,partial cell organs manifest side-gathering phenomenon on chromatin,and the link between SW480 cell is loosened , subsequently partial rounded cell is floated.When the H2RA(cimetidine 40μM、nizatidine 20μM) against SW480 cells after 3 day,SW480 cells change roundly,nucleolus smashed,shrinking nucleolus came into being,and part of those not adhibited cell wall.

在光镜和倒置相差显微镜下,与对照组相比,经姬姆萨染色观察,西咪替丁、尼扎替丁处理的SW480细胞,先发生皱缩,体积变小,异形性减少,细胞变圆数目增多,细胞核变小,染色变淡,核仁数量减少,部分可见染色质边集现象;细胞间连接疏松,随后部分变圆细胞浮起。40μM西咪替丁及20μM尼扎替丁连续处理3天后,SW480细胞全部变圆,细胞核碎裂,固缩微核形成,部分不再贴壁。

The hilling activing of CD3AK+ cells showed significantly higher to K562/VCR cells than to K562 cells, while there was no difference of cytotoxicity between K562 /HHT and K562 cells.

4CD3AK+细胞对K562/VCR细胞的杀伤活性强于对敏感细胞K562,其对K562/HHT细胞的杀伤活性与其对K562细胞的杀伤活性比较,差异无显著性。

Concludes:Three subsets of mast cell in human tissue are sorted and depurated with antibodies of specific enzyme and immunofluorescence labelling and flow cytometry. Mast cell of containing profuse secretary vacuoles could be displayed by confocal microscopy. It demonstrate that mast cell have material substructure offering for fast reaction of I type allergies of human being.

利用肥大细胞的特征性酶抗体、免疫荧光标记和流式细胞仪可将人组织中的肥大细胞分选纯化为三种亚型;以共聚焦显微镜显示肥大细胞含有丰富的分泌颗粒,它说明肥大细胞具备了为人体I型变态反应提供快速反应的物质基础。

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