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201.
AIM To construct the mouse embryonic stem cell (ESC) line with stable pancreatic and duodenal homeobox 1 (Pdx1 ) expression by Tet-On system, which may lay a foundation for further research on the differentiation of Pdx1+ definitive endoderm cells into pancreatic cells. METHODS The Pdx1 -overexpressing lentiviral vector with green fluorescent protein marker and puromycin resistance was constructed by Tet-On system and was used to infect the mouse ESC. The cells were divided into 3 groups: blank control group (ESC group), empty lentivirus control group (PDX1- ESC group) and Pdx1 lentivirus transfection group (PDX1+ ESC group). Flow cytometry was used to detect the transfected cells after screening by doxycycline (DOX). The function of Tet-On system and the expression of Pdx1 gene were detected. The transfected cells in PDX1- ESC group and PDX1+ ESC group were sorted by flow cytometry, and constructed ESC line with stable expression of Pdx1 and negative control ESC line were verified. RESULTS (1) The positive rates of transfected cells in PDX1- ESC group and PDX1+ ESC group were 90.72% and 94.01% after screening by DOX, respectively. The positive rates of transfected cells in PDX1- ESC group and PDX1+ ESC group was 97.84% and 98.13% after sorting by flow cytometry, respectively. (2) With DOX, green fluorescence was observed in PDX1- ESC group and PDX1+ ESC group. The mRNA and protein expression of Pdx1 was significantly increased in PDX1+ ESC group (P <0.05). Without DOX, no green fluorescence was observed in the cells of the 3 groups, and no significant difference in the mRNA and protein expression of Pdx1 was observed (P >0.05). (3) After 3 months of cryopreservation, the cell lines still survived in resuscitation culture and were regulated by DOX. CONCLUSION Using Tet-On system, the mouse ESC line with inducible Pdx1 expression were successfully established and could be used as an effective cell model to research the differentiation of Pdx1+ definitive endoderm cells into pancreatic cells. 相似文献
202.
选择了4种家鱼作指示生物,用紫外分光光度法测定其白肌中RNA和DNA的含量。结果表明:同种鱼,污染区的RNA含量显著低于对照区,白鲢和鳊鱼白肌中RNA/DNA低于对照区;不同种的鱼,污染区鱼肌核酸比对照区下降的程度有鳊鱼>白鲢>鲫鱼和鲤鱼的趋势。鱼肌核酸下降同水体重金属污染有一定联系,RNA是比RNA/DNA更合适的指标,鳊鱼和白鲢是比鲫鱼和鲤鱼更敏感的指示生物。 相似文献
203.
经电镜扫描寡齿新银鱼的第Ⅳ时相,第Ⅴ时相卵母细胞和受卵的卵细胞粘丝,结果表明,卵细胞表面具从受精孔发生的12根左右粘丝,此粘丝于受精孔处微弱弯曲,经侧部渐向受精孔相对处呈现很强扭曲或互相缠绕。卵细胞粘丝不仅出现在受精卵和第Ⅴ时相卵母细胞表面,而且清晰地出现在第Ⅳ时相卵母细胞的滤泡膜上。 相似文献
204.
大银鱼卵细胞粘丝扫描电镜观察 总被引:1,自引:0,他引:1
用扫描电镜观察了大银鱼的第Ⅲ时相,第Ⅳ时相,第Ⅴ时相卵母细胞和受精卵的卵细胞粘丝的外部形态特征和它们的粘性,并讨论了卵细胞粘丝的功能。 相似文献
205.
206.
在棉花根冠细胞生物测定过程中,棉花黄萎病病菌通过提纯复壮而达到较强的侵染能力,并制备出较多的黄萎病菌粗毒素。介绍了粗毒素的简易制备、根冠细胞培养与制备过程以及染色过程中染色剂的选择、染色时间的长短。实验与传统方法相比有几点改进:总结出细胞振荡时间在1~2 min之内;之后黑暗条件下静置培养2~6 h;选用pH为6.5的0.01%中性红染色3~5 min,之后加入一滴0.2%伊文思兰染色,伊文思兰的染色时间应严格掌握在3min以内等,为根冠细胞测定方法的推广提供了一定的理论支持。 相似文献
207.
两种菊酯类农药对鲤血细胞的影响 总被引:2,自引:0,他引:2
采用对鲤肌肉注射不同浓度的高效反式氯氰菊酯和功夫菊酯,研究了两种菊酯类农药对鲤血细胞形态、红细胞渗透脆性的影响。结果显示:随两种农药攻毒浓度增大、时间延长,鲤血细胞异形率增大。低浓度(6、60μg/kg)中毒鲤成熟红细胞大小不一,核稍有畸形,淋巴细胞、单核细胞增多,核浆界限不清,有空泡,甚至伴有核溶解的白细胞;而高浓度(600、6 000μg/kg)中毒鲤血细胞中除伴有低浓度中毒症状,还出现了染色质模糊,部分白细胞核浆界限不清,甚至溶解,退化更明显。红细胞渗透脆性亦随攻毒浓度增大、时间延长而增高,且开始溶血点的变化幅度较完全溶血点大,表明最小抵抗值更易受环境因子影响。 相似文献
208.
209.
Eui Jeong Han Hyun-Soo Kim Kalu Kapuge Asanka Sanjeewa Kyungsook Jung Youngheun Jee You-Jin Jeon Ilekuttige Priyan Shanura Fernando Ginnae Ahn 《Marine drugs》2020,18(12)
Sargassum horneri (S. horneri), an edible brown alga, has been proposed as a functional food with an improvement effect on abnormal skin immune responses. The present study investigates the anti-allergic effect of an ethanol extract from S. horneri (SHE) on immunoglobulin E (IgE)/bovine serum albumin (BSA)-mediated activation in bone marrow-derived cultured-mast cells (BMCMCs) and passive cutaneous anaphylaxis (PCA) reaction in mice. SHE markedly and dose-dependently suppressed the degranulation of BMCMCs by reducing the β-hexosaminidase and histamine release without cytotoxicity. In addition, SHE significantly decreased the FcεRI expression on the surface of BMCMCs and its IgE binding. Moreover, SHE reduced the mRNA expression and the production of allergic cytokines; interleukin (IL)-1β, IL-4, IL-5, IL-6, IL-10, IL-13; interferon (IFN)-γ and/or tumor necrosis factor (TNF)-α; and a chemokine, thymus and activation-regulated chemokine (TARC), by suppressing the activation of Src-family kinases and nuclear factor (NF)-κB signaling. In further study, the application of SHE reduced the PCA reaction in an IgE/BSA-induced type I allergic mice model. Taken together, we suggest that SHE has an anti-allergic effect in type I allergic responses. 相似文献
210.
ZHENG Hao-long SONG Nan CAO Hui-min CHEN Si WANG Jie Lü Xiao-ming JIA Lian-qun 《园艺学报》2020,36(4):713-718
AIM To investigate the effects of different components of Gynostemma pentaphyllum [gypenosides (Gps), gypenoside XLIX (GpXLIX) and ginsenoside Rb3 (GRb3)] on mitochondrial energy metabolism-related proteins in endothelial cells induced by oxidized low-density lipoprotein (ox-LDL). METHODS EA.hy926 cells were divided into control group, model group, Gps group, GpXLIX group and GRb3 group. The cells in control group were cultured only in DMEM complete medium. The cells in model group were treated with 100 mg/L ox-LDL for 48 h. The cells in Gps group, GpXLIX group and GRb3 group were treated with 100 mg/L ox-LDL for 24 h, and then treated with Gps, GpXLIX and GRb3 at 100 mg/L for another 24 h, respectively. The ATP content in each group was detected by ELISA. The expression levels of mitochondrial energy metabolism-related proteins, cytochrome C oxidase subunit 5a (Cox5a), NADH:ubiquinone oxidoreductase core subunit S1 (Ndufs1), ATP synthase F1 subunit alpha (ATP5a) and cytochrome C (Cyt C), were determined by Wes automatic Western blot quantitative analysis system and Western blot. RESULTS Compared with control group, the ATP content in model group was decreased (P <0.01). After drug intervention, the ATP content increased to different degrees in Gps group, GpXLIX group and GRb3 group (P <0.01). The results of Wes automatic Western blot quantitative analysis system were consistent with those of Western blot. These results showed that compared with control group, the protein expression of Cox5a, Ndufs1 and ATP5a in model group was decreased, and the protein expression of Cyt C was increased (P <0.01). After intervention, the protein expression of Cox5a, Ndufs1 and ATP5a was increased and the protein expression of Cyt C was decreased in Gps group, GpXLIX group and GRb3 group (P <0.05 or P <0.01). Among them, the effect of Gps on the protein expression of Cox5a, Ndufs1 and Cyt C was significantly stronger than those of the 2 monomer components, and the effect of GRb3 was found to be superior in the 2 monomer components. The effect of GpXLIX on ATP5a protein was superior to the other 2 components. CONCLUSION Gynostemma total saponins and related active ingredients protect ox-LDL-induced endothelial cells by affecting mitochondrial energy metabolism-related proteins, thereby preventing and treating atherosclerosis. 相似文献