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3Chondrogenesis was assessed using Alcian blue staining, Toluidine bluestaining, Safranin O/Fast Green staining at 7 and 14 days, still collagenⅡimmunohistochemistry and aggrecan immunofluorescence at 4, 7 and14 days after initial chondrogenic induction of adipose -derivedmesenchymal stem cells.

3观察脂肪间充质干细胞在诱导后的生长增殖情况、形态变化,于诱导7、14天后应用阿尔新兰染色、甲苯胺兰染色、藩红0/固绿染色,以及4、7、14天后应用Ⅱ型胶原蛋白免疫细胞化学、aggrecan免疫荧光评价其成软骨能力。

The extracellular matrix wasstained positively for Alcian blue, Toluidine blue and Safranin O / FastGreen at 7 and 14 days after initial chondrogenic induction of adipose -derived mesenchymal stem cells, and indicated collagenⅡand aggrecanwere expressed positively.

2脂肪间充质干细胞在高密度的"微团"培养条件下,于诱导7、14天后阿尔新兰染色、甲苯胺兰染色、藩红0/固绿染色阳性,诱导4、7、14天后Ⅱ型胶原蛋白免疫细胞化学、aggrecan免疫荧光阳性表达。

E and Sirus red in supersaturated picric acid solution;the amount of HA in serum was measured with the method of radiative immunity;the quantitative expression of fibrosis factor—TGF、PDGF was measured with the method of immunity histochemistry.Results:Myocardial collagen in model group increased obviously compared with normal group from the results of electron microscope、H.E、Sirus red in supersaturated picric acid solution.

腹腔注射柯萨奇B3病毒建立病毒性心肌炎慢性期心肌纤维化小鼠模型,随机分为模型对照组、通心络低剂量组、中剂量组、高剂量组,并设正常对照组,用药四周后处死动物通过透射电镜观察心肌细胞超微结构;HE染色、苦味酸天狼星红染色观察心肌胶原数量;放免法测定血清透明质酸的含量;免疫组化染色观察致纤维化细胞因子—转化生长因子和血小板衍生生长因子的定量表达。

Methods:Establish the model of myocardial fibrosis in chronic stage of viral myocarditis by injecting CVB3 into Balb/c rats,divide these rats into model group、Tong xin luo low dose group、Tong xin luo medium dose group、Tong xin luo high dose group and set up normal group,after four weeks of giving medicine execute the animals and observe the ultrastructure of myocardial cell with tramission electron microscope,the quantity of myocardial collagen was observed by staining with H.E and Sirus red in supersaturated picric acid solution;the amount of HA in serum was measured with the method of radiative immunity;the quantitative expression of fibrosis factor—TGF、PDGF was measured with the method of immunity histochemistry.

腹腔注射柯萨奇B3病毒建立病毒性心肌炎慢性期心肌纤维化小鼠模型,随机分为模型对照组、通心络低剂量组、中剂量组、高剂量组,并设正常对照组,用药四周后处死动物通过透射电镜观察心肌细胞超微结构;HE染色、苦味酸天狼星红染色观察心肌胶原数量;放免法测定血清透明质酸的含量;免疫组化染色观察致纤维化细胞因子—转化生长因子和血小板衍生生长因子的定量表达。

Then the content of the interleukin (IL-1) in serum were measured by the method of enzyme-linked assay; From the HE dyeing, the change of morph in the arthrodial was observed by the light microscope; From the AB-PAS dyeing, the change of PG in the cartilage matrix was observed; By all of that the therapeutic efficacy of the medicine wine Yong GuKangNing to the disease of osteoarthritis was estimated, and the mechanisms to the disease was also investigated.

用酶免法测定血清中白细胞介素(IL-1)的含量;常规HE染色,光镜下观察关节软骨结构形态学变化;AB-PAS染色,光镜下观察软骨基质中蛋白聚糖的染色变化;评价永康宁骨康宁对膝骨性关节炎的治疗效果,并探讨其作用机制。

The location and structures of sex-pheromone-producing gland in female H.insularis were studied by EAG,GC,SEM,and TEM.These studies showed that thegland situate in the intersegmental membrane between the eighth and ninthabdominal segments,and is an eversible abdominal fold;Many plump cones disturbon the surface of the gland.The glandular cells of 2-day old virgin female H.insularis are arranged in one layer,among which the central cells are columnarepithelial cells and flat on two sides.The nucleus is irregular elliptical.There isevident conjugation between cells and the involution is more in the basal membraneof cell.Microvilli are distributed on the cytoplasmic membrane and linked withendocuticle on which there are many layers of chitin,and the outer cuticule is staineddeeper.The cell contains bubbles,mitochondria,glycogen deposits,roughendoplasmic reticulum and smooth endoplasmic reticulum.

结合触角电位、毛细管气相色谱、扫描电镜、透视电镜等技术对小线角木蠹蛾雌蛾腹尖末端不同组织部位提取物的测定分析以及腺体位置和形态结构的观察发现:小线角木蠹蛾性信息素分泌腺位于腹部末端8~9节之间,是一个由节间膜特化而成的上皮结构,为一可外翻的腹褶,腺体表面分布着饱满的锥形体,羽化后2天未交尾的雌蛾腺体细胞呈单层排列,腹面中央由密集的柱形上皮细胞组成,细胞排列向两侧延伸至背部,其形状由柱形逐渐变为扁平形,细胞核为椭圆形,细胞与细胞间有明显的胞连接,细胞基底膜基褶较多,质膜上分布着微绒毛,并与内表皮连接,内表皮上有多层几丁质,外角质层染色较深,细胞质中含有空泡、线粒体、脂质粒、粗面内质网和光面内质网。

Bone marrow-derived mesenchymal stem cells are capable of chondrogenesis, making them a possible source of cells for injectable cartilage tissue engineering. There exist different ideas on the ability of mesenchymal stem cells's chondrogenesis in monolayer culture. Because of this, the effect of adult rabbit's bone marrow-derived mesenchymal stem cells chondrogenesis in monolayer culture was studied. The mesenchymal stem cells was isolated from adult rabbit's bone marrow and monolayer cultured. TGF-β1, Vit-C and Dexamethasone were used. Immunohistochemistry analyses and histological staining of H-E, Methylaniline blue and Alcian blue were performed to identify the expression of collagen type Ⅱ and cartilage associated matrix. The results showed that the induced cells expressed and produced collagen type Ⅱ and cartilage associated matrix. This suggests that the differentiation of adult rabbit's marrow-derived mesenchymal stem cells into chondrocyte in monolayer culture is feasible and may be induced by TGF-β1, Vit-C and Dexamethasone.

骨髓基质干细胞的软骨分化潜能使其可能成为可注射组织工程化软骨研究的种子细胞,为探讨体外培养的骨髓基质干细胞在平面诱导条件下软骨分化的可行性,我们进行下列实验:获取并体外平面培养成体兔骨髓基质干细胞,应用TGF-β〓、Vit-C和地塞米松对其软骨分化诱导,诱导后的骨髓基质干细胞行细胞爬片组织学和Ⅱ型胶原免疫组化,结果证实,诱导后骨髓基质干细胞可分泌Ⅱ型胶原,组织学染色可见类似于软骨细胞,由此证明体外培养的骨髓基质干细胞在平面诱导条件下可以软骨分化,其软骨诱导因子为TGF-β〓、Vit-C和地塞米松。

MAIN OUTCOME MEASURES: The morphology of BMSCs was observed by the inverted phase contrast microscope. Expression of BMSC surface antigens was detected by flow cytometer. Osteogenous potential was assessed by alkaline phosphatase staining. Adipogenic potential was evaluated by Oil red O staining. Chondrogenic potential was assessed by toluidine blue staining.

主要观察指标:倒置相差显微镜下观察骨髓间充质干细胞的生长状态,流式细胞仪检测细胞表面标志物的表达,采用碱性磷酸酶染色鉴定成骨能力,以油红O染色鉴定成脂能力,以甲苯胺蓝染色鉴定成软骨能力。

In Part 1 of our research, BGSCs were sorted through immunomagnetic beads marking by CD133 and cultured in vitro, and character as a stem cell was identified by stem cell markers (CD133 and Nestin) and differentiated cell markers [ microtubule-associated protein 2(MAP2), glial acidic fibrillary protein and myelin basic protein] , ultrastructure observing with electron microscopeand engrafting to severe combined immunodeficiency mice for tumorigenesis test. The results were as following: Only a small subset of CD133+ glioma cells in glioma cell lines and fresh specimens from various pathologic grade could express stem cell markers CD133 and Nestin, view ultrastructure of a stem cell and be capacity of serial passage in culture. These CD133+ cells possese a marked capacity for multipotent differentiation and could differentiate into tumor cells expressing MAP2,β-TubulinⅢ, GFAP and MBP; When engrafted into SCID mice, they can generate and form tumors that phenotypically resembl the tumor from the patient.

在本课题中,在第一部分实验中采用以CD133为标志的免疫磁珠法从人脑胶质瘤组织和细胞株中分离脑胶质瘤干细胞并进行体外培养,通过免疫荧光技术检测干细胞标志物CD133、Nestin,诱导分化后检查分化细胞标志物MAP2、GFAP、MBP以及电镜超微结构观察和移植SCID鼠致瘤实验,对其干细胞特性加以鉴定,得到如下结果:不同病理分级的新鲜胶质瘤标本和胶质瘤细胞株中存在一小部分CD133+的胶质瘤细胞,能表达干细胞的标志物CD133和Nestin,符合干细胞的超微结构特点,体外培养能连续传代;具有多向分化潜能:诱导分化后能产生MAP2、β-TubulinⅢ、GFAP、MBP染色阳性的细胞;移植SCID鼠后能形成与亲本肿瘤表型一致的移植瘤。

Histologic examination revealed pleomorphic cellular proliferation with xanthomatous, multi and mono nucleated giant cells.

免疫组织化学染色显示瘤细胞胶质纤维酸性蛋白阳性,网质纤维染色显示单个瘤细胞间着色。

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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.

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