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Methods CD34(superscript +) cells and AC133(superscript +) cells were gathered by MiniMACS, then the cell factors were divided into 6 groups. Through the ex vivo comparison cultivation under different cell factor combination stimulation, expansion of the CD34(superscript +) and AC133(superscript +) selection cells was observed and the clone formation of the CD34(superscript +) and AC133(superscript +) cells was also observed by using Half-solid Methyl cellulose under different cell factor stimulation (during 7d, 14d, 21d.). The rate of cell apoptosis was determined.

常规富集人骨髓CD34及AC133细胞,应用本研究组设计并已经证实的细胞因子组合方式,对人骨髓CD34及AC133富集细胞进行体外对照培养,分别观察CD34和AC133富集细胞的扩增情况;应用甲基纤维素半固体培养法,观察不同组别培养7、14、21d CD34和AC133富集细胞的集落形成情况及细胞凋亡率。

There is not a significant difference in these groups by one-way analysis of variance. 2 Among 51 cases with lung cancer, 7 specimens showed malignant cells in bronchial lavages, including 4 cases in central lung cancer and 3 cases in peripheral lung cancer. Concordant results were observed in 20 cases. 6 specimens showed malignant cells on cytological analysis, and NMSP was positive in at least one gene tested. 14 samples did not show malignant cells on cytological analysis, and the NMSP results were correspondingly negative. The results of cytology and NMSP were discordant in 31 samples. 30 samples were cytologically negative, but NMSP positive in one or more genes. In addition, there was one case that the cytology was positive for malignant cells, but the NMSP was negative in all of the three gene tested.

51例肺癌患者支气管灌洗液细胞学检查发现肺癌细胞7例,其中27例中心型肺癌患者4例支气管灌洗液发现肺癌细胞,24例周围型肺癌患者3例支气管灌洗液发现肺癌细胞。20例支气管灌洗液细胞学检查和过甲基化检测得到一致结论。6例支气管灌洗液发现肿瘤细胞,过甲基化分析至少一个基因阳性。14例灌洗液未发现肿瘤细胞,三个基因过甲基化分析均为阴性。31例支气管灌洗液细胞学检查和过甲基化检测得到不一致结论,30例细胞学检查为阴性,但过甲基化分析一个或一个以上基因为阳性。1例细胞学检查阳性但过甲基化分析阴性。

Interleuk in 2 (IL-2) is an important cytokine released from activated T lymphocytes, which activated lymphokine actived killer cells and natural killer cells , and promoted cytokine production of helper T Cells and natural killer cells , and stimulated T cell growth in vitro and so on . It plays a important role in body immune regulator.

白细胞介素2主要是由激活的T淋巴细胞产生的一类重要的细胞因子,具有激活淋巴因子活化的杀伤细胞和自然杀伤细胞,刺激T辅助细胞和自然杀伤细胞产生细胞因子和促进细胞因子的繁殖以及刺激T细胞在体外生长等功能,因此它是在免疫调节中有重要作用。

At the same time, the process of the wound healing was observed histologically, and the regeneration cycle of epidermal cells and the temporal change in inflammatory cells were measured. Results Inflammatory cells infiltrated into wound surface were neutrophils, followed by macrophages and lastly lymphocytes. Epidermal cells proliferated most actively on PBD 3 and the mitoses of them increased significantly on day 7 after burn. The gene expression of PDGF, PDGFR and EGFR reached peaks on PBD 1 and the gene expression of EGF and TGFβ-R2 were highest on PBD 3. In addition, the gene expression of TGFβ-R1 and TGFβ1 increased significantly on PBDs 5 and 7 respectively.

结果 炎性细胞到达创面依次为中性粒细胞、巨噬细胞和淋巴细胞;表皮细胞周期变化为伤后3天细胞增殖活跃,伤后7天细胞分裂明显增多;伤后1,3天PDGF、PDGFR(血小板源性生长因子受体)和EGFR基因表达明显增强,伤后3,5天EGF、TGFβ-R2基因表达增强,伤后5,7天TGFβ-R1(转化生长因子β受体1)基因表达增强,伤后7,10天TGFβ1基因表达增强。

Objective:The aim of the experiment is to search for the regulation that the hepatic stem cells abnormally differentiate into cancer cells. In the experiment,the variating regulation of PCNA,c-kit and c-myc in the formation and development of HCC will be interpreted with immunohistochemistry and RT-PCR. So,the initial mechanism that hepatic stem cells abnormally differentiate into cancer cells from the levels of cell,protein and gene will be clarified. The last,the inhibition to HCC with ulinastain will be studied,from which,a new way of clinical prevention and treatment for HCC will expect to be explored.

二、目的本实验旨在探讨以卵圆细胞为代表的肝干细胞在HCC发生、发展过程中在肝组织内分布规律,并通过免疫组化、RT-PCR等技术阐明PCNA、c-kit、c-myc在HCC发生、发展过程中的动态演变规律,从细胞、蛋白、基因三个水平初步研究卵圆细胞向癌细胞异常分化的机制,同时观察乌司他丁对HCC发生、发展的抑制作用,为肝癌的临床预防和治疗探索新的途径。

By incorporation of tritiated thymidine or 5-bromodeoxyuridine and using them as the markers, we found that the basal cells, Clara cells and type II alveolar epithelial cells in the pulmonary tissues may in some way behave like stem cells and can eventually differentiate into respiratory tract epithelium.

通过应用cells in induced group cultured for 2 weeks 标记的胸腺嘧啶(3H-TdR)或5-溴脱氧尿核苷的渗入进行标记,发现肺组织的基底细胞、Clara 细胞和肺泡II型上皮细胞可能具有干细胞特征,能最终分化形成气道上皮。

By incorporation of tritiated thymidine or 5-bromodeoxyuridine and using them as the markers, we found that the basal cells, Clara cells and type Ⅱ alveolar epithelial cells in the pulmonary tissues may in some way behave like stem cells and can eventually differentiate into respiratory tract epithelium.

通过应用cells in induced group cultured for 2 weeks von kossa staining)标记的胸腺嘧啶(3H-TdR或5-溴脱氧尿核苷的渗入进行标记,发现肺组织的基底细胞、clara细胞和肺泡Ⅱ型上皮细胞可能具有干细胞特征,能最终分化形成气道上皮。

The result showed that the surface architecture and ultrastructure of I.punctatus ' gill and secondary gill lamellae were similar to those of other teleosts. The surface of gill arches and gill rakers was covered with pavement cells which were characterized by circular microridges, and secreting-holes were found among cells on the surface of gill arches. The base part of gill rakers were bigger in terms of diameter. The pavement cells' surface of gill filaments' middle part was characterized by microridges with fine lateral protuberances on both sides which were either fused or interruped, while the microridges disappeared on the pavement cells' surface on both top and base parts of gill filaments, forming dense granulated protuberances. The height and interlamellar space of secondary gill lamellae in I.punctatus were relatively higher and bigger, respectively.

结果表明,斑点叉尾鮰鳃的表面结构和微细结构与其他硬骨鱼类基本相似,鳃弓和鳃耙表面被具环形微嵴的扁平上皮细胞所覆盖,鳃弓表面细胞之间有孔洞;鳃耙基部较粗壮;中部鳃丝上皮细胞表面的微嵴两侧有细小的横突,有些微嵴出现融合或间断;端部和基部鳃丝上皮细胞表面的微嵴消失,形成密集颗粒状突起;鳃小片的高度和片间距较大。

Results In spheroids culture system, under the condition of apoptosis of src-transformed cells, the apoptosis of chemical transformed cells were also induced by normal cells, but BPV-transformed cells were resistant.

结果:在球体细胞培养实验系统中,在src转化细胞产生凋亡的条件下,甲基胆蒽转化细胞同样能产生凋亡,而BPV转化细胞则抵抗了由周围正常细胞所致的凋亡。

Methods Marrow-derived mesenchymal stem cells were isolated and purified from mouse femoral bone and shinbones using differential adherent methods. Cells at the third passage were induced by 20% FBS in conditioned medium, conditioned medium alone, 20% FBS or 10% FBS alone respectively. Mouse aortic smooth muscle cells were cultured as the positive control. Levels of mRNA and protein expression of myoeardin and several smooth muscle cells marker genes were determined by immumofluorescenee, RT-PCR and Western blot before and 3, 7, 10, 14d after the induction. The presence of smooth muscle myofilaments was detected by using transmission electron microscope.

采用伞骨髓贴壁法分离骨髓问充质干细胞,CM联合20%FBS诱导骨髓间充质干细胞,同时设立持续10%FBS、单20%FBS及单CM诱导下的骨髓间充质十细胞对照组和SMC阳性对照组,分别于诱导前及诱导3、7、10和14 d时观察细胞形态的变化,并在相应的时间点用免疫荧光法、逆转录聚合酶链反应法、Western blot半定量分析法榆测myocardin以及SMC表面各种标志基因的表达变化,用透射电镜检测诱导后细胞内肌丝存在以此来证实诱导分化成功。

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