ABSTRACT
Objective:To explore the feasibility of direct renin inhibitor aliskiren for the treatment of severe or critical coronavirus disease 2019 (COVID-19) patients with hypertension.Methods:The antihypertensive effects and safety of aliskiren was retrospectively analyzed in three severe and one critical COVID-19 patients with hypertension.Results:Four patients, two males and two females, with an average age of 78 years (66-87 years), were referred to hospital mainly because of respiratory symptoms. Three were diagnosed by positive novel coronavirus 2019 (2019-nCoV) nucleic acid or antibody, and the critical patient with cardiac insufficiency was clinically determined. Two patients were treated with calcium channel antagonist (CCB), one with angiotensin converting enzyme inhibitor (ACEI), and one with angiotensin Ⅱ receptor antagonist (ARB). After admission, ACEI and ARB were discontinued, one patient with heart failure was treated by aliskiren combined with diuretic.Three patients were treated with aliskiren combined with CCB among whom two withdrew CCB due to low blood pressure after 1 to 2 weeks. Based on comprehensive treatment including antiviral and oxygenation treatment, blood pressure was satisfactorily controlled by aliskiren after three to four weeks without serious adverse events. All patients were finally discharged.Conclusion:Our preliminary clinical data shows that antihypertensive effect of aliskiren is satisfactory and safe for severe COVID-19 patients complicated with hypertension.
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.
Subject(s)
Humans , Adenocarcinoma , Pathology , Cell Line, Tumor , Cell Movement , Cell Proliferation , Cyclin-Dependent Kinase Inhibitor p21 , Genetics , Metabolism , Jumonji Domain-Containing Histone Demethylases , Pharmacology , Lung Neoplasms , Pathology , RNA, Messenger , Genetics , Metabolism , TransfectionABSTRACT
To investigate the effect and mechanism of NK-1 Tachykinin receptor (NK-IR) antagonist on hypoxia induced hepatic injury, we established the hypoxic rat model. 30 male SD rats (weighing 240-300g) were randomly divided into 3 groups, control group, and experimental groups including the hypoxia group and the NK-1R antagonist group. The rats of experimental groups underwent hypoxia, among them the NK-1R antagonist group were those with interference of NK-1R antagonist by intraperitoneal injection. Hepatic injury was evaluated by pathological staining, hepatic function detection and hepatocellular apoptosis determination. Results showed hypoxia-induced hepatic injury in rats was established successfully. Edema,ballooning degeneration and spotty necrosis were found in livers in the experimental groups, among which the pathological injury in the hypoxia group was worse than that in the NK-1R antagonist group. Moreover,GGT and the rate of hepatocellular apoptosis in the NK-1R antagonist group were obviously lower than that in the hypoxia group (P<0.05). But no significant difference were found in ALT,AST and ALP between groups (P>0.05). These data indicate that Substance P possibly participate in the process of hypoxia-induced hepatic injury, and NK-1R antagonist could reduce hypoxia-induced hepatic injury.
Subject(s)
Animals , Male , Rats , Apoptosis , Hepatocytes , Pathology , Hypoxia , Pathology , Liver , Liver Function Tests , Neurokinin-1 Receptor Antagonists , Random Allocation , Rats, Sprague-DawleyABSTRACT
Hypoxia-inducible transcription factor-1alpha (HIF-1alpha) is a "master" gene in the cell hypoxia response network. HIF-1alpha plays the key role on VEGF-dependent angiogenesis during tumor development. In this study, the siRNA expressive vector targeting HIF-1alpha was constructed. HepG2 cells were tranfected with an HIF-1alpha siRNA vector and exposed to a normoxic or hypoxic environment. mRNA and protein levels of HIF-1alpha and VEGF were detected by RT-PCR and Western blot at different cell culture environment. The mRNA and protein expression of HIF-1alpha was inhibited by RNA interference. The obvious decreases of mRNA and protein of VEGF was induced by HIF-1alpha gene knock-down. Our study indicated that HIF-1alpha regulated the expression of VEGF and hence promoted angiogenesis in liver cancer under hypoxic environment. The expression of VEGF was downregulated via HIF-1alpha gene knock-down. These results suggest that HIF-1alpha may be used as a good target gene for anti-angiogenesis in liver tumors.