ABSTRACT
The aim of this study was to observe whether necroptosis is involved in the process of cardiac hypertrophy induced by pressure overload. SD rats underwent transverse abdominal aortic constriction (TAC) operation for establishing cardiac hypertrophy model. The structure and function of the left ventricle of rats were evaluated via echocardiography, left ventricular mass index, the expression of markers of cardiac hypertrophy and histological detection. Real-time PCR and Western blot were used to measure the gene and protein expression of receptor interacting protein kinase 1 and 3 (RIPK1 and RIPK3, the necroptosis markers) respectively. Four weeks after TAC operation, rat model for cardiac hypertrophy was established. The experimental data showed that the gene and protein expressions of RIPK1 and RIPK3 in the rat heart hypertrophic tissues after TAC for 4 weeks were increased significantly compared with those in the sham group. HE staining showed cardiomyocytes injury and hypertrophy in the hearts of TAC rat models. By transmission electron microscope, we observed that mitochondria of cardiomyocytes were damaged seriously in the TAC models. Treatment with losartan used, the selective antagonist of angiotensin II type I receptor could improve the cardiac function of TAC rats. Moreover, losartan treatment decreased the expression of RIPK1 and RIPK3 in heart tissues of TAC rats. The results suggest that necroptosis occurrs in the process of cardiac hypertrophy with pressure overload, and losartan could alleviate the cardiac hypertrophy and inhibit necroptosis.
Subject(s)
Animals , Rats , Apoptosis , Cardiomegaly , Pathology , Disease Models, Animal , Echocardiography , Heart , Losartan , Pharmacology , Myocytes, Cardiac , Pressure , Protein Serine-Threonine Kinases , Metabolism , Rats, Sprague-Dawley , Real-Time Polymerase Chain Reaction , Receptor-Interacting Protein Serine-Threonine Kinases , MetabolismABSTRACT
Objective To exam the personality traits and self-concept of school phobia patients with different academic achievements.Methods 38 school phobia patients with high academic achievement and 19 school phobia patients with low academic achievement were assessed with Eysenck' s personality questionnaire and Piers-Harris Children' s Self-concept Scale.Results High academic achievement group of psychoticism and neuroticism higher than that of low academic achievement group.High academic achievement group of Self-concept's total scores,behavior,intelligence and the school situation higher than that of low academic achievement group.High academic achievement group of anxiety scale score lower than that of low academic achievement group.The personality traits of school phobia correlated with the self-concept on some level.Conclusion Different academic achievements of school phobia patients have different personality traits and have different self-concept,so they need to individualized treatment options.
ABSTRACT
This study aims to construct the plasmid of human Runx1 and observe its possible effects on Runx1 gene expression in human pulmonary adenocarcinoma cells (A549). The shRNA sequence targeting human Runx1 was designed and synthesized, then inserted into pSuper plasmid by DNA recombination technology. The recombinant plasmid was confirmed by bacterial colonies PCR, enzyme digestion analysis and DNA sequencing. A549 cells were transfected by Runx1 shRNA plasmid. The inhibition efficiency of pSuper-Runx1-shRNA plasmid on Runx1 at mRNA level and protein level were measured with real-time PCR and Western blot. The results of real-time PCR and Western blot indicated that the mRNA and protein levels of Runx1 in A549 cells were inhibited by the pSuper-Runx1- shRNA expression plasmid, and the inhibition rate were 33% and 50%, respectively.
Subject(s)
Humans , Cell Line, Tumor , Core Binding Factor Alpha 2 Subunit , Genetics , Gene Expression , Genetic Vectors , Plasmids , Genetics , RNA, Messenger , Genetics , RNA, Small Interfering , Genetics , Real-Time Polymerase Chain Reaction , TransfectionABSTRACT
This study was aimed to construct transgenic mouse model with target for Runxl gene. Runxl cDNA of mice was amplified by PCR from pcDNA3. 1 Flag Runx1 FL vector and inserted into ptetO7-Asc-IRES-EGFP vector to form a recombinant vector, and then the recombinant vector was injected into fertilized egg by microinjection technology to get a transgenic mouse. The results of PCR and Southern blot indicated that the Runx1 transgenic mouse was constructed successfully, and this could provide an important tool for studying the function of Runxl gene in vivo.
Subject(s)
Animals , Female , Male , Mice , Base Sequence , Core Binding Factor Alpha 2 Subunit , Genetics , Genetic Vectors , Genetics , Mice, Inbred C57BL , Mice, Transgenic , Genetics , Microinjections , Molecular Sequence Data , Recombination, GeneticABSTRACT
For investigating the effect of Jumonji domain-containing protein-3 (JMJD3) on the behavior of lung cancer cell line, A549 proliferation was measured with EDU staining and flow cytometer after JMJD3 expression plasmid and pcDNA3. 1 transfection at 48h. The migration ability of A549 was tested at the same time. The expression of p21 mRNA was measured with RT-PCR. The results showed that JMJD3 transfection increased the EDU positive cells ratio (JMJD3: 40.75% +/- 2.07%, control: 20.97% +/- 1.5%, P < 0.001). G1 phase cell ration also increased after JMJD3 transfection (JMJD3:47. 80% +/- 1.85%, control: 54.60% +/- 0.95%, P = 0.005). The mRNA expression of p21 decreased in JMJD3 group (JMJD3: 35. 89% +/- 3.71%, control: 91.78% +/- 3.74%, P < 0.001). The distances of migration were (0.47 +/- 0.27) cm and (0.96 +/- 0.40) cm after 24h and 48h with JMJD3 tranfection, compared to (0.57 +/- 0.22)cm and (1.08 +/- 0.33)cm in control, respectively (P > 0.05). JMJD3 promoted the proliferation of A549 and decreased the G1 cell numbers, decreased the p21 mRNA, but had no effect on A549 migration.