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成纤维细胞

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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。

Methods: The effects of Opuntia on proliferation of fibroblast were measured with MTT colorimetric assay, and PCNA productions were examined with immunocytechemistry.

以小鼠成纤维细胞株(NIH3T3)为研究对象,采用四氮唑盐比色法测定仙人掌对成纤维细胞增生的影响,采用免疫细胞化学染色法检测成纤维细胞的增殖细胞核抗原的表达。

In order to investigate the inhibitory effect of salvia miltiorrhiza and tetramethyl pyrazine on scartricial fibroblast, the hypertrophic scar tissue of chest was chosen for culture of fibroblasts, and the influence of SM and TP on fibroblasts was observed, The effect of the drugs on the growth of fibroblasts, on DNA synthesis of fibroblasts and on mitosis index of fibroblasts were all determined quantitatively.

摘 要 增生性瘢痕是临床治疗难题之一,目前尚无有效的治疗方法。为了探讨丹参和川芎嗪对瘢痕成纤维细胞的抑制作用,取12例患者胸部增生性瘢痕组织,通过体外成纤维细胞培养,探讨不同药物浓度、不同作用时间对成纤维细胞生长的影响。

When electric fusion method was used for nuclear transfer, the fusion rate (46. 0%), cleavage rate (53. 9%) and blastocyte development rate (10.9%) of adult ear fibroblasts were significantly lower than that of fetal fibroblasts (64. 5%, 70.1%, 21. 6% respectively), fetal skin cells (71. 5%, 70.8%, 22. 1% respectively) and ovary granulosa cells (88. 2%, 79. 1%, 25. 5% respectively). There was no significant difference among other donor cells in the cleavage and blastocyst development rate of resconstituted embryos.

当用电融合法进行核移植时,成体耳部成纤维细胞的融合率(46.0%),卵裂率(53.9%)和囊胚发育率(10.9%)均显著低于胎儿成纤维细胞(64.5%,70.1%和21.6%),胎儿皮肤细胞(71.5%,70.8%和22.1%),以及卵巢颗粒细胞(88.2%,79.1%和25.5%);另外三种细胞间的卵裂率,囊胚发育率无显著差异,但卵巢颗粒细胞的融合率显著高于胎儿成纤维细胞和胎儿皮肤细胞(88.2%vs 64.4%,71.5%,P<0.05)。

Special staining methods, such as Masson and the Van Gieson staining were used to study the distribution of collogen fibers and elastic fibers. ResultsBy HE staining, the subepithelial connective tissues and vessels in the pterygium were more prominent than normal conjunctival tissues. An amorphous subepithelial superficial hyalinized zone and coarse eosinophilic granular materials were observed in the pterygia, but they were not found in normal conjunctival specimens. Coarse fibers were visible only in the deeper subepithelial connective tissues of pterygial samples. With Masson′s staining, the dense staining of collagen fibers was also more prominent in the pterygium than in the subepithelial connective tissues of normal conjunctiva. Abnormal collagen fibers were visible in the deeper sub-epithelial connective tissues of pterygial samples. With Van Gieson staining, abnormal collagen fibers were visible in the deeper subepithelial connective tissues. Dark coarse elastic fibers were found in the abnormal fibers only in the subepithelial deep connective tissues of pinguecula in the pterygia but not in the conjunctiva. With immunohistochemistry staining, MMP-3 was strong in the pterygial epithelium, moderate in fibroblast and absent from pterygial vascular walls. LN was strongly expressed in the blood vessel wall, moderately in the epithelial basement membrane and absent from the entire stroma.

结果HE染色:翼状胬肉组织上皮下基质中存在结缔组织的增生和血管形成;基质浅层存在一无定形物质透明区及粗糙的颗粒样嗜酸性物质,在翼状胬肉体部深层基质中存在粗糙的纤维组织;正常球结膜组织细胞排列整齐;基质为疏松结缔组织,胶原纤维平行排列,其间可见成纤维细胞,散在少量中性粒细胞、毛细血管;Masson染色:翼状胬肉浅层基质中存在致密的胶原纤维染色,深层基质中的胶原纤维存在变性样改变;VG染色:翼状胬肉组织深层基质中存在大量变性的胶原纤维,其间夹杂黑色的弹性纤维;免疫组化染色法:MMP-3在翼状胬肉上皮细胞中呈强表达,成纤维细胞中呈中等强度表达,血管内皮细胞中未见表达;LN在血管壁中呈强表达,在上皮细胞基底膜中呈中等强度表达,在整个基质中未见明显表达;col Ⅲ在整个翼状胬肉基质中呈强表达。

Results The results showed that fibroblasts were the main source of extracellular matrix production in bile duct wall. The phenotype of fibroblasts in inflammatory strictured bile duct wall changed obviously, quiescent fibroblasts were activated and transformed to myofibroblasts, with massive proliferation.

结果 成纤维细胞是胆管壁中合成胶原的主要细胞;炎性狭窄胆管壁的纤维化增厚明显,其成纤维细胞表型改变,由相对静止转化为功能活跃状态,并进一步向肌成纤维细胞转化,且大量增殖。

Objective: To explore the mechanisms resulting in the recurrence of urethral scar which make urethral strictures difficult to be cured, a series experiments were conducted to find potential effective factors involved in urethral scar formation and degradation, including the studies of extracellular matrix component of urethral stricture scar, the characteristics of urethral scar fibroblast, and the effects of urine on urethral fibroblast in vitro, as well as the studies to compare the difference of collagenase activity, type Ⅰ collagen and tissue inhibitor of metalloproteinase-1 (TIMP-1) in the tissues and cultured fibroblasts from normal urethra and strictured urethra respectively, and the studies to investigate the effect of antisense TIMP-1 oligodeoxyonucleotide on cell proliferation and collagenase activity of urethral scar fibroblast.

中文题名尿道瘢痕基础研究副题名胶原酶活性,TIMP-1的表达及其反义基因治疗外文题名 Experimental study on urethral scar-activity of collagenase,expression of TIMP-1,and antisense TIMP-1 gene transfection of urethral scar fibroblast 论文作者黄翔导师杨宇如教授学科专业外科学研究领域\研究方向学位级别博士学位授予单位四川大学学位授予日期2002 论文页码总数104页关键词尿道手术瘢痕胶原酶成纤维细胞尿道瘢痕馆藏号BSLW /2003 /R699 /12 目的:研究尿道瘢痕的细胞外基质的组成,尿道瘢痕成纤维细胞的生物学特性以及尿液对其生长的影响;比较胶原酶活性,金属蛋白酶组织抑制因子-1(TIMP-1)以及Ⅰ型胶原含量在尿道瘢痕和正常尿道组织及体外培养的成纤维细胞中的差异;研究反义TIMP-1寡核苷酸对尿道瘢痕成纤维细胞增殖以及胶原酶活性的影响。

Results: Compared with control group, apoptosis cells increased from 0.5%to 10%(some even to 15%) after 24,48 and 72 h action of -300,-500 and -1 000 V electrets. After action of -500 V PTFE electrets for 48-72 h, fibroblast cells showed characteristic morphological features of apoptosis. These features included chromatin aggregation, nuclear and cytoplasmic condensation and partition of cytoplasm and nucleus into membrane bound-vesicles.

结果:-300、-500和-1 000 V驻极体作用成纤维细胞24、48和72 h以后,与对照组相比,成纤维细胞的凋亡量从0.5%增至10%(部分可达15%);驻极体作用成纤维细胞48~72 h,出现细胞凋亡特有的形态学特征,即:细胞异染色质边集,细胞裂解,可见凋亡小体。

To understand the infectivity by porcine endogenous retrovirus with porcine skin fibroblast cell in vitro and in vivo, porcine skin fibroblast cell established by our laboratory were co-cultured with neo/HEK293 cell for the infection of RERV in vitro, and were subcutaneously transplantated to SCID (severe combined immuno-deficiency) mice for the infection of PERV in vivo, laying the foundation for valuation of biologic safety of xenotrans-plantation. The event of neo/HEK293 cells infected by PERV occurred during co-culture of porcine skin fibroblast cells with neo/HEK293 cells, expanding the rang of the infection of porcine endogenous retrovirus. Afterpig cells transplantated subcutaneously in SCID mice, the microchimerism (78.57%) of pig cells occurred widel, and there was phenomena of integration of PERV provirus (85.71%) in several organs or tissues remote from the injected sites, indicating infection of PERV in SCID mice in vivo. yet, there is no evidence of active viral replication in analysis of PERV env RNA of these tissues or organs.

为了解猪皮肤成纤维细胞PERV在体外和体内的感染性,通过建立猪皮肤成纤维细胞系,将所建细胞系与人胚胎肾293细胞体外共培养,并移植于严重联合免疫缺陷鼠皮下进行猪皮肤成纤维细胞PERV的体外和体内感染性实验,结果表明,猪皮肤成纤维细胞与人胚胎肾细胞共培养过程中,猪内源性逆转录病霉感染人胚胎肾细胞,进一步证实和拓宽了猪细胞PERV感染人细胞的范畴;猪皮肤成纤维细胞移植SCID鼠皮下后,导致SCID鼠发生猪细胞微嵌合(78.57%)和PERV在体内感染(85.71%)并且波及远离移植部位的多种组织或器官,但是并未检测出SCID鼠组织中表达PERV env RNA。

In order to confirm this suspect, we choosed a cultured rat granulation tissue fibroblasts model in vitro and applied RT-PCR and cell-ELISA technology to detect the changes of firbroblasts intrinsic EGF, bFGF, TGFβ-1 with their receptors gene expressions and protein synthesises after stimulated by SP. Our purpose was to explore the possible effects and patterns imposed by SP on fibroblasts intrinsic growth factors and their receptors expressions, which maybe offer theoretical basis for promoting wound healing via improving nervous functions and regulating neuropeptides secretions.

为进一步验证这一推测,同时排除在体多因素干扰,我们采用了一种大鼠肉芽组织成纤维细胞体外培养模型,采用RT-PCR与细胞ELISA技术,检测SP刺激成纤维细胞后,成纤维细胞内源性EGF、bFGF、TGFβ-1及其受体基因表达和蛋白合成的改变情况,探讨SP对成纤维细胞内源性生长因子及其受体表达的影响以及方式,以期为经由改善神经功能、调节神经肽分泌途径促进伤口愈合提供理论依据。

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