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Results: As compared with those of the irradiation control groups, the rhCuZnSOD showed significant beneficial effects on immunocytes, these included enhancing the proliferative response of T lymphocyte to concanavalin A and the activity of natural killer cells in spleen, increasing the number of whole spleen lymphocyte, leucocyte and lymphocyte counts in peripheral blood.

与照射对照组比较,rhCuZnSOD可减轻电离辐射对小鼠免疫细胞的影响,其作用包括增强脾T淋巴细胞对刀豆蛋白A诱导的增殖反应,提高脾自然杀伤细胞的活性,升高全脾淋巴细胞及外周血白细胞和淋巴细胞的数量。

Apoptosis of lymphocytes in bone marrow, spleen, thymus and bursa was detected by light microscope, electron microscope and terminal- deoxynucleotidyl transferase UIP nick end labeling. The results showed ALV-J could induce incoordinate apoptosis of lymphocytes in thymus, bursa and spleen, which was mainly found in early time. And the apoptosis was much more obvious in thymus than in other immune organs.

以电镜观察和HE染色、原位末端标记染色光镜观察对ALV-J感染和混合感染鸡的骨髓、脾脏、胸腺、法氏囊等免疫器官中淋巴细胞的凋亡进行了形态学研究,首次发现ALV-J可诱导胸腺、法氏囊和脾脏中不同程度的淋巴细胞凋亡,凋亡主要见于早期,ALV-J与REV混合感染时淋巴细胞凋亡尤为显著,以胸腺中最为明显。

Objective To detect the expression of CD70 in peripheral T lymphocytes of patients with systemic lupus erythematosus and the effect of azacitidine, an inhibitor of DNA methylation, on it. Methods Blood samples were obtained from 10 patients with active SLE (SLEDAI score ≥5), 10 patients with nonactive SLE (SLEDAI score 5) and 10 normal human controls.

目的 通过研究共刺激分子CD70在SLE患者外周血T淋巴细胞的表达水平,以及DNA甲基化抑制剂氮杂胞苷对正常人外周血T淋巴细胞表达CD70分子的影响,探讨DNA甲基化在SLE患者外周血T淋巴细胞表达CD70分子中的调控作用。

Get high purity DCs by Cultured plastic-adherent monocytes isolated from healthy human peripheral blood with GM-CSF and IL-4 for 7 days. To observe the morphology of DCs by inverted phase contrast microscope ,electron microscope and laser confocal microscope. Analyse phenotype of DCs with flow cytometry. Investigate the endocytosis ability of DCs as a group by Horseradish peroxidase endocytosis assay. To appraise allogeneic mixed lymphocytes reaction of DCs by MTT reduction assay. Analyse the levels of IL-12 and TNF in liquids of cultured medium by ELISA and MTT reduction assay respectively. Soluble antigens of HCCs was obtained by 3 freeze-and-thaw cycles. Biological characteristics of HC soluble antigens pulsed DCs were monitored by flow cytometry. According to MTT reduction assay estimated the cell proliferation of self lymphocytes activated by HC antigens pulsed DCs. Get high purity BCG HSP 70 protein by SDS-PAGE electrophoresis and determined its biological activity with ELISA. Analyse phenotype of antigen pulsed DCs primed by BCG HSP70 with flow cytometry. By MTT reduction assay estimated the cell proliferation of self lymphocytes and the MLR of DC based vaccine. Analyse expression of HLA-DR molecule on surface of HCC lines. The IFN-γ mRNA in lymphocytes after actived by DC vaccine and the Fas-L expression on DC and DC vaccine primed lymphocytes were detected by in situ hybridization and flow cytometry respectively. Specific cytotoxity lysis of T lymphocytes and nonspecific inhibition of liquids in culture medium against HCC lines were also tested. Detect expression of hAFP on four HCC lines with Cell-ELISA. Induce apoptosis of HCCs with actinomycin-D. Interaction of DCs and apoptotic cells was observed under transmission electron microscope. Growth inhibition test of DC against HCC lines was also performed. Establish the nude mouse model bearing human HC xenografts and indentify the characteristic of tumour by histochemistry and immunohistochemistry techniques. Prevent and treat transplanted human HC on nude mouse with Freezing and anabiotic HC specific lymphocytes.

用GM-CSF和IL-4从健康人外周血诱导DC;分别用倒置相差显微镜、电子显微镜及激光共聚焦显微镜观察DC形态;流式细胞术检测DC表型;HRP吞噬实验测定DC的群体内吞能力;MTT法检测同种异体混合淋巴细胞反应;ELISA法和MTT法分别测定DC培养上清液中IL-12和TNF水平;冻融法制备肝癌细胞可溶性抗原;流式细胞术检测负载肝癌可溶性抗原后DC的生物学特性;MTT法检测DC负载肝癌抗原后对自身淋巴细胞增殖的影响;SDS-PAGE制备电泳纯化BCG HSP70并鉴定纯度,ELISA测定活性;流式细胞术检测负载抗原DC经BCGHSP 70活化后的表型;MTT法检测肝癌DC疫苗对自身淋巴细胞增殖的影响和混合淋巴细胞反应;流式细胞术检测肝癌细胞表面HLA-DR表达;MTT法检测肝癌DC疫苗对自身淋巴细胞的活化;原位杂交法检测肝癌DC疫苗活化后的淋巴细胞IFN-γmRNA表达;流式细胞术检测DC和肝癌DC疫苗活化后淋巴细胞表面Fas-L;MTT法分别检测肝癌DC疫苗活化的淋巴细胞和其培养上清对肝癌细胞的特异性杀伤和非特异性抑制作用;Cell-ELISA检测人肝癌细胞hAFP表达;MTT法检测负载AFP表位肽和凋亡肝癌细胞DC对自身淋巴细胞增殖的影响;ELISA法和MTT法分别测定活化后淋巴细胞培养上清中TNF和IL-12水平;肝癌细胞凋亡的诱导和检测;DC吞噬凋亡肝癌细胞后的电子显微镜观察;DC对肝癌细胞的生长抑制试验;人肝癌裸鼠皮下移植瘤动物模型的建立及其组织学和免疫组织化学鉴定;DC及肝癌特异性淋巴细胞预防和治疗人肝癌裸鼠皮下移植瘤;冻存和复苏后的肝癌特异性淋巴细胞预防和治疗人肝癌裸鼠皮下移植瘤。

The results of study show:1. White pulp and red pulp in parenchyma of spleen of embryo can be obviously discerned after 18 days. Periarterial lymphoid sheath and ellipsoid periarterial lymphoid sheath also can be obviously discerned in spleen of 4 days chicken. T, B lymphocytes in appendix basement of embryo emerge after 20 days. It is the initial shape of cecal tonsil. Crypt structure of conjunction of esophago and stomachus glandularis form obviously at 4 days. It is the initial shape of esophago tonsil. The germinal center firstly emerges in these three organs at 14 days. With the increase of day age, the characteristic structure peripheral immune organs gradually develop mature. Spleen achieve mature at 21 days and cecal tonsil at 35 days.2. IgM~+ and IgA~+ cells in spleen of embryo emerge at 15 days. IgG~+ cell, CD3~+ and CD8~+T lymphocytes of embryo emerge at 20 days. CD3~+, CD8~+ and IgM~+ cells in cecal tonsil of embryo emerge at 20 days. However CD4~+, IgG~+ and IgA~+ cells all emerge in 1 day age chicken out of crust. CD3~+, CD4~+, CD8~+, IgM~+ and IgG~+ cells in esophago tonsil of embryo all emerge at 20 days. However IgA~+ cells emerge in 1 day age chicken out of crust.3. The amount of T, B lymphocytes in peripheral immune organs increase follow with the increase of day age, and hold an upgrade tendency. The amount of T, B lymphocytes in spleen achieved stabilization at 21 days, and in tonsil of esophago and appendix at 35 days.

研究结果表明:1、在组织结构方面,脾脏实质内的白髓与红髓在胚胎18日龄后明显可辨,4日龄雏鸡脾脏中形成明显可辨的动脉周围淋巴鞘和椭球周围淋巴鞘;盲肠基部T、B淋巴细胞在胚胎20日龄时开始出现,即初步形成盲肠扁桃体;食管与腺胃结合处在4日龄时形成明显的隐窝结构,即食管扁桃体初步形成;14日龄时,三种器官中首次出现生发中心;随着日龄的增长,外周免疫器官特征结构不断发育成熟,脾脏在21日龄时达到成熟水平,盲肠扁桃体和食管扁桃体在35日龄时达到成熟水平。2、在T、B淋巴细胞出现时间方面,脾脏中IgM~+和IgA~+细胞在胚胎15日龄时开始出现,IgG~+细胞、CD3~+和CD8~+T淋巴细胞在胚胎18日龄时出现,CD4~+细胞在胚胎20日龄时出现;盲肠扁桃体中CD3~+、CD8~+和IgM~+细胞在胚胎20日龄时开始出现,而CD4~+、IgG~+和IgA~+细胞均在雏鸡出壳后1日龄时出现;食管扁桃体中CD3~+、CD4~+、CD8~+、IgM~+和IgG~+细胞均在胚胎20日龄时开始出现,而IgA~+细胞则在雏鸡出壳后1日龄时出现。3、在T、B淋巴细胞数量变化方面,外周免疫器官中T、B淋巴细胞的数量随日龄增长,整体均呈上升趋势。

To evaluate the efficacy of the recombinant fowlpox virus vaccine against the heterotype IBV, the chickens were challenged with the homotype IBV LX4 strain and the heterotype IBV LTJ95I strain after three weeks post vaccination. Antibodies against IBV were detected in vaccinated chickens one week post inoculation. The CD4+ T-lymphocytes in the peripheral blood increased rapidly in all groups challenged with IBV, except for the vaccinated group challenged by heterotype strain and the low level of CD4+ T-lymphocytes remained until end of the experiment. In all the groups, a high level of CD8+ T-lymphocytes only was observed in the vaccinated group after challenging with IBVhomotype strain. The morbidity and the mortality of this group were 21.43 % and 0 %, respectively, which showed significant difference with other groups. In addition, the lesions of chickens and virus shedding were less in the vaccinated group challenged by IBV homotype strain comparing with other groups, but there was no difference for the average body weight of chickens in all groups.

结果显示,重组疫苗接种1周后,免疫鸡产生抗IBV的抗体;而且外周血中CD4+和CD8+ T淋巴细胞的含量略高于非免疫对照组;攻毒后,异源强毒株攻毒的免疫组CD4+ T淋巴细胞呈下降趋势,并且该组低水平CD4+的状态一直持续到试验结束,而其他组CD4+ T淋巴细胞均迅速上升,峰值达到14.5 %;同源强毒株攻毒的免疫组CD8+ T淋巴细胞呈高水平的表达,而其他组攻毒后均无明显变化;保护率结果显示,同源强毒株攻毒免疫组的发病率和死亡率为21.43 %和0 %,与其他各组相比均有显著差异;另外同源强毒株攻毒的免疫组病理损伤与异源强毒株攻毒的试验组相比明显减轻,其排毒时间和排毒量也均有所减少;强毒攻毒后所有试验组体重无显著差异。

The results were that PNWⅠ had a significant effect at the concentration of 200μg/mL and PNM had no significant effect on the splenocyte proliferation.

初步考察了PNW和PNWⅠ对小鼠体外淋巴细胞增值的影响,结果显示PNWⅠ在200μg/mL时对淋巴细胞有显著的促进增殖作用,PNW对淋巴细胞的增殖作用不显著。

Objective To establish a method for isolating and culturing the tonsillar lymphocytes,analyze the distribution of lymphocyte phenotypes so as to get some experimental data for peripheral lymphocyte research.

目的 建立人扁桃体淋巴细胞的分离培养方法,研究淋巴细胞的表型分布,为外周淋巴细胞的研究提供实验数据。

Methods The human lymphocytes were isolated from the tonsillar tissues of the patients having a clear clinical diagnosis of tonsil sleep apnea.

目的 建立人扁桃体淋巴细胞的分离培养方法,研究淋巴细胞的表型分布,为外周淋巴细胞的研究提供实验数据。

Results The activation of NF-κB, proliferation response, and expression of IL-4 and I L-5 mRNA and protein in T lymphocytes stimulated by PMA were significantly hig her than those of their blank control (P.01), while those indexes of T l ymphocytes stimulated by PMA and PDTC simultaneously were significantly lower th an those stimulated by PMA alone (P.01). The apoptotic index of T lympho cytes stimulated with PMA were significantly lower than that of their blank cont rol (P.01), and the apoptotic index of asthmatic guinea pig T lymphocytes stimulated with PMA and PDTC simultaneously were significantly higher than that stimulated by PMA alone (P.01). The significant positive correlations w ere found between the activation of NF-κB and the proliferation (r=0.64, P.001), and the expression of IL-4 and IL-5 mRNA and protein of T lymph ocytes, respectively (r=0.55-0.68, P.001). There was also signific ant negative correlation between the activation of NF-κB and apoptosis of T l ymphocytes (r=0.62, P.001). Conclusions NF-κB may participate in the signal conduction of PKC regulated proliferation, apoptosis and expression of IL-4 and IL-5 of T lymphocytes in asthma.

结果加入PMA培养的哮喘组T淋巴细胞NF-κB的活化、细胞增殖反应、IL-4和IL-5的mRNA和蛋白质的表达均显著高于其空白对照(P.01),而同时加入PMA和PDTC培养的哮喘组T 淋巴细胞以上指标均显著低于只加入PMA培养的哮喘组T淋巴细胞(P.01);加入PMA 培养的哮喘组T淋巴细胞的凋亡指数显著低于其空白对照(P.01),而同时加入PMA 和PDTC培养的哮喘组T淋巴细胞凋亡指数显著高于只加入PMA培养的哮喘组T淋巴细胞(P 。01)。T淋巴细胞NF-κB的活化与增殖反应呈显著正相关(r=0.64,P.00 1),与IL-4和IL-5的mRNA和蛋白质的表达也均呈显著正相关(r=0.55-0.68,P 。001),而与凋亡指数呈显著负相关(r=-0.62,P.001)。

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