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TSPG has the similar effect as HGFs, can directly or/and indirected effect and regulate proliferation and differentiation of CD34〓 HSC/HPC to bring into full play in"invigorating qi"of Panax ginseng. 2. During the process of the extensive proliferation, orderly differentiation, functional activation and gene transfer of hematopoietic stem/progenitor cells in ex vivo culture systems, TSPG may be used to regulate the proliferation and differentiation of hematopoietic cells to perform potential application in the domain of hematpoietic stem cell engineering.

我们的研究结果提示:1、TSPG具有类似造血生长因子的功效,能直接或间接影响和调控造血干/祖细胞的增殖与分化,促进血细胞的生成,从而发挥人参&补气生血&之功效。2、在进行对造血干/祖细胞的扩增与诱导分化,功能激活,基因转染等造血干细胞工程研究中可利用人参皂甙的药理作用来调控细胞的增殖分化,发挥其在造血干细胞工程领域的潜在应用价值。

To sum up, both experiments in vivo and in vitro show that activating blood circulation to dissipate blood stasis has effects of inhibiting proliferation of malignant cell, inducing differentiations and impelling apoptosis. These perhaps were the machinisms of clinical effects. Considering the effects on DNA content, we dares inferred that YDQ inhibited proliferation by restraining the synthesis of DNA. It asks for our doing more work to research the underlying machinism with immunology, cytobiology, molecularbiology and so on.

总之,体内、体外实验均表明活血化瘀为主的疗法对恶性细胞有一定的增殖抑制、分化诱导和凋亡促进作用,这也许正是临床治疗MDS取效的原因所在,分析瘀毒清对细胞DNA含量的影响,我们推测认为瘀毒清可能通过抑制细胞DNA的合成进而抑制恶性细胞的增殖,其更深的机制尚待我们从免疫学、细胞和分子生物学等角度进一步研究揭示。

Both Fas and NO can lead chondrocyte apoptosis respectively and cause articular cartilage destruction. IGF-Ⅰ, TGF-β, bFGF, BMP and other growth factors are polypeptide agents that can influence cell activity by signal convection. They can accelerate chondrocyte proliferation and proteoglycan synthesis, play the local regulation action on formation and plerosis of bone and cartilage tissue by autocrine or paracrine. They have the ability to induce cartilage formation. Some investigations showed that growth factors can influence chondrocyte metabolism, synthesis of specific Ⅱ type collagen and proteoglycan by co-operation and inhibition. 1. 3 Situation of OA therapeutics The therapeutic methods of OA mainly comprised non-drug treatment, drug treatment, operation treatment, tissue and genetic engineering, et al. Drug treatment is the chief method among them.

若其活性发生改变,则将导致关节软骨基质成分的丢失和进行性破坏;软骨细胞凋亡与OA的发病密切相关,Fas与NO可各自独立介导软骨细胞凋亡,造成关节软骨破坏;IGF-Ⅰ、TGF-β、bFGF、BMP等生长因子是一组通过细胞间信号传递影响细胞活动的多肽因子,具有促进细胞生长、增殖与合成等作用,可通过自分泌或旁分泌方式对骨和软骨的形成和修复起局部调节作用,可促进软骨细胞增殖、分化与蛋白多糖的合成,具有较强的诱导软骨形成的能力,研究表明多种生长因子相互促进、相互抑制,以协同或拮抗方式影响软骨细胞代谢,影响软骨细胞特异性Ⅱ型胶原和蛋白多糖的合成分泌。

The human cartilages are composed of chondrocyte and extracellular matrix , the form of chondrocytes are hypertrophy and the quantity are less; the ECM of cartilage are compised of type Ⅱ collagen and proteoglycan. Articular cartilages are all hyaline with little fibers. Trauma and arthritis are the main cause of cartilage injury, the ommilayer injury ofcartilage can be recovered by marrow, but because of without stimulation mechanism, the new tissues are merely fibrocartilages, they can not be coincide with hyaline cartilage in menchanics; the purely damage of articular cartilage can not stimulate chondrocyte to regenerate because of without blood circulation, thus, the plerosis of articular catilage can not depend on the proliferation of local chondrocyte. Ever since, people tried their best to find a way to reconstruct articular cartilage.

中文题名人骨髓基质干细胞成软骨诱导及多孔复合材料作为细胞载体的体外实验研究副题名外文题名 Cartilage induction of human mesenchymal stem cells and experiment on compound porous materials as cells' scaffold in vitro 论文作者刘晓岚导师周江南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中南大学学位授予日期2003 论文页码总数68页关键词骨组织工程软骨细胞骨髓基质干细胞壳聚糖高分子外消旋聚乳酸馆藏号BSLW /2003 /R68 /10 造成人体关节软骨损伤的原因主要为创伤和关节炎,关节软骨全层损伤可由于骨髓中间充质干细胞的高速增殖修复,但这种修复由于缺乏相应的刺激机制,只能形成纤维软骨,而不能形成符合关节生理、力学要求的透明软骨;单纯软骨部分损伤软骨组织内无血管,软骨细胞迁移迟缓,无法使损伤区域软骨细胞再生,因此,关节炎及关节创伤后的软骨修复不能依赖于软骨细胞的增殖和迁移。

Of CBMC can be purified by Mini-MACS as CD34〓 stem cells. B. The number of CD34〓 stem cells can expand to 40.24±9. 86 fold after 14 days. C. No matter in the expression of CD1a, CD80, CD86, and HLA-DR, or in the function of stimulating xenogenous lymphocyte proliferation, there was no difference between CD34+ stem cell DCs or monocyte DCs. D. The percentage of CD3〓CD56〓 cells is the same in CIK cells co-culture with DCs transfected with SKOV3 RNA, CIK cells co-culture with DCs, and CIK cells. E. The expansion rate of CIK cells can be accelerated by co-culturing with loaded or unloaded DCs. However, the expansion rate between loaded or unloaded group is the same. F. The strongest cytotoxicity against SKOV3 cell line was achieved by CIK cells co-cultured with DCs loaded with SKOV3 RNA.

结果:1、Mini-MACS分选系统可自CBMC中提取0.78±0.31%的CD34〓细胞;2、体外培养14天后可获得原始CD34〓细胞量40.24±9.86倍的细胞;3、不论在CD1a、CD80、CD86及HLA-DR的表达上,或是刺激异体淋巴细胞增生的功能上,脐带血CD34〓细胞与单核细胞来源的DC都没有差别;4、CIK细胞中CD3〓CD56〓双阳性的表达率在与RNA转染DC共培养的CIK细胞组、与DC共培养的CIK细胞组及单纯CIK细胞组3组间比较无差异;5、脐带血CIK细胞增殖显著,培养14天时可扩增18.18±5.59倍,培养21天时可扩增35.02±6.30倍;5、与未转染或转染DC共培养的CIK细胞在培养第14天后增殖速率大于单纯CIK细胞。

The Food and Drug Administration of USA hasapproved the use of this topical medicine for treatment of different malignant andpremalignant tumors such as actinic keratoses and basal cell carcinoma. Althoughimiquimods exact mechanisms of action in neoplastic diseases has not yet been fullyelucidated, animal and human studies have shown its ability to enhance the productionof cytokines such as interferon-, interleukin-12, and tumor necrosis factor- though conjugations with Toll like receptors on the surface of dendriticcells of the monocyte-macrophage lineage.

有关咪喹莫特抗癌作用的具体机制仍不十分明了,一般认为,咪喹莫特主要是通过与免疫相关细胞表面的Toll样受体结合、促进IFN-α、TNF-α、IL-12等多种前炎症细胞因子的分泌,进而激活天然与获得性免疫反应的方式发挥抗癌作用;进一步的研究发现,咪喹莫特尚能选择性的抑制某些皮肤肿瘤细胞的增殖并诱导其凋亡,提示增殖抑制和诱导凋亡可能是评价咪喹莫特抗肿瘤作用的重要指标和机制。

The proliferative capacity of LEC from human had directly relevance to the age of donor(r=-0.996).Conclusions The formation of lens-like spherules is characteristics of LEC lines in the cultured cells.Under identical conditions,the proliferative rate of LEC from bovine and rabbit is fast than that of LEC from human,but dedifferentiation of LEC from bovine and rabbit is easier than that of LEC from human;LEC from the three species exhibit similar limited growth potential.The proliferative rate of LEC from human has a inversely proportion with age.

&晶状体小体&的形成可作为确定晶状体上皮细胞株的一项特征性依据,而体外培养的人、牛、兔晶状体上皮细胞具有相同的有限生长潜能,在相同的条件下,牛、兔晶状体上皮细胞的生长增殖速度比人晶状体上皮细胞快,但易于发生去分化;此外,人晶状体上皮细胞的生长增殖率与年龄密切相关,年龄越小,晶状体上皮细胞的生长增殖速度越快。

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。

Meanwhile,HDAC1 acetylation level is significantly increased.The overexpression of HDAC1 can promote erythroid cell proliferation and inhibits induced differentiation,the knockdown of HDAC1 or 2 inhibits erythroid cell proliferation and promotes induced differentiation.We also found that HDAC1 affects GATA1 mediated transcription activity.Thus our data revealed that HDAC1 may negatively regulate GATA1 mediated transcription and suggest that the dynamic regulation of GATA1 associated NuRD/MeCP1/HDAC1/HDAC2 complex may determine the onset of erythroid differentiation programs.Moreover,this study will eventually help to design new therapeutic approaches in treatment of leukemia.

研究发现在被诱导分化的小鼠红细胞白血病细胞中与GATA1相结合的HDAC1被乙酰化并失去活性,在血细胞中过表达HDAC1能够促进血细胞的增殖并抑制分化,反之在血细胞中敲除HDAC1或HDAC2能抑制血红细胞的增殖并促进分化,而且HDAC1能够负调控GATA1的转录活性,GATA1与NuRD/MeCP1复合体之间的动态调控,决定血红细胞分化的发生和发展,并为探索白血病的致病机理奠定了理论基础。

Result: With a high cancer-inhibiting rate on the animal model having been transplanted the cancer, this medicine can remarkably restrain the growth of cancer cells cultivated in vitro and improve the life quality of mouse model, and also shows obvious promotion and enhancement function on the DTH ear-swelling of mouse with cancer, the proliferation of mouse spleen lymphocyte induced by ConA and the cytokine (TNF-α、IFN-γ) of mouse with cancer respectively. It can obviously decrease the adverse effects like weight reduction, damage of viscera and degradation of life quality caused by the chemotherapeutic medicines, and has a remarkable stimulation on the lymphocyte proliferation induced by ConA. It is also able to improve the ratio of SMMC-7721 cells in G0-G1 period and reduce the cell amount in G2/M period to cut down the possibility of stepping into the division period; additionally, it can obviously inhibit some cancer genes of the cancer cells and meanwhile up regulate the cancer-inhibiting gene.

结果:本品能明显抑制体外培养肿瘤细胞的生长,对在体肿瘤移植模型动物具有较高的抑瘤率,能明显提高模型小鼠的生存质量;对荷瘤小鼠DTH耳肿胀、ConA诱导的小鼠脾淋巴细胞增殖、荷瘤小鼠细胞因子(TNF-α、IFN-γ)呈现明显的促进或提高作用;能显著减少化疗药所引起的小鼠体重降低、脏器损害以及生存质量下降等不良作用;本品含药血清对ConA诱导的淋巴细胞增殖具有明显的促进作用;能提高SMMC-7721细胞G0-G1期比例,降低G2/M期细胞数目,而减少其进入分裂期;对肿瘤细胞的多个癌基因具有明显的抑制作用,同时上调抑癌基因等。

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