Tannic acid alleviates lipopolysaccharide­induced H9C2 cell apoptosis by suppressing reactive oxygen species­mediated endoplasmic reticulum stress.

Sepsis­induced myocardial dysfunction is one of the features of multiple organ dysfunction in sepsis, which is associated with extremely high mortality and is characterized by impaired myocardial compliance. To date, there are few effective treatment options available to cure sepsis. Tannic acid (TA) is reportedly protective during sepsis; however, the underlying mechanisms by which TA protects against septic heart injury remain elusive. The present study investigated the potential effects and underlying mechanisms of TA in alleviating lipopolysaccharide (LPS)­induced H9C2 cardiomyocyte cell apoptosis. H9C2 cells were treated with LPS (15 µg/ml), TA (10 µM) and TA + LPS; control cells were treated with medium only. Apoptosis was measured using flow cytometry, reverse transcription­quantitative PCR (RT­qPCR) and western blot analysis. Additionally, the levels of cellular reactive oxygen species (ROS), malondialdehyde and nicotinamide adenine dinucleotide phosphate were evaluated. Western blotting and RT­qPCR were also employed to detect the expression levels of endoplasmic reticulum (ER) stress­associated functional proteins. The present findings demonstrated that TA reduced the degree of LPS­induced H9C2 cell injury, including inhibition of ROS production and ER stress (ERS)­associated apoptosis. ERS­associated functional proteins, including activating transcription factor 6, protein kinase­like ER kinase, inositol­requiring enzyme 1, spliced X box­binding protein 1 and C/EBP­homologous protein were suppressed in response to TA treatment. Furthermore, the expression levels of ERS­associated apoptotic proteins, including c­Jun N­terminal kinase, Bax, cytochrome c, caspase­3, caspase­12 and caspase­9 were reduced following treatment with TA. Additionally, the protective effects of TA on LPS­induced H9C2 cells were partially inhibited following treatment with the ROS inhibitor N­acetylcysteine, which demonstrated that ROS mediated ERS­associated apoptosis and TA was able to decrease ROS­mediated ERS­associated apoptosis. Collectively, the present findings demonstrated that the protective effects of TA against LPS­induced H9C2 cell apoptosis may be associated with the amelioration of ROS­mediated ERS. These findings may assist the development of potential novel therapeutic methods to inhibit the progression of myocardial cell injury.

Inhibition of long noncoding RNA MALAT1 suppresses high glucose-induced apoptosis and inflammation in human umbilical vein endothelial cells by suppressing the NF-κB signaling pathway.

The study investigated the expression of long noncoding RNA (lncRNA) MALAT1 in high glucose (HG)-induced human vascular endothelial cells (HUVECs) and the role of MALAT1 in the apoptosis of HG-induced HUVECs. The HUVECs were cultured and induced with 25 mmol/L HG. After that, the HUVECs were transfected with MALAT1 siRNA. The expression levels of MALAT1 were detected with qPCR, whereas the expression levels of Bax, Bcl-2, cleaved-caspase-3, cleaved-caspase-9, p-65, and p-p65 were detected using Western blot. The roles of MALAT1 in cell activities, including apoptosis, were evaluated using the CCK-8 assay, TUNEL staining, and flow cytometry. The expression levels of inflammatory factors (TNF-α and IL-6) were measured using ELISA. The expression levels of MALAT1, TNF-α, and IL-6 in HUVECs were increased in the HG environment; however, when MALAT1 was silenced in the HUVECs, cell proliferation increased significantly, the expression levels of TNF-α, IL-6, Bax, cleaved-caspase-3, and cleaved-caspase-9 decreased, and the rate of apoptosis also decreased. Silencing MALAT1 inhibited the expression of p-p65 in HG-induced HUVECs. In conclusion, our study demonstrated that MALAT1 is upregulated in HG-induced HUVECs, and inhibition of MALAT1 inhibits HG-induced apoptosis and inflammation in HUVECs by suppression of the NF-κB signaling pathway.

Identification of MSX1 and DCLK1 as mRNA Biomarkers for Colorectal Cancer Detection Through DNA Methylation Information.

Colorectal cancer is the second most deadly malignancy in the United States. However, the currently screening options had their limitation. Novel biomarkers for colorectal cancer detections are necessary to reduce the mortality. The clinical information, mRNA expression levels and DNA methylation information of colorectal cancer were downloaded from TCGA. The patients were separated into training group and testing group based on their platforms for DNA methylation. Beta values of DNA methylation from tumor tissues and normal tissues were utilized to figure out the position that were differentially methylated. The expression levels of mRNA of thirteen genes, whose CpG islands were differentially methylated, were extracted from the RNA-Seq results from TCGA. The probabilities whether the mRNA was differentially expressed between tumor and normal samples were calculated using Student's t-test. Logistic regression and decision tree were built for cancer detection and their performances were evaluated by the area under the curve (AUC). Twenty-four genomic locations were differentially methylated, which could be mapped to eleven genes. Nine out of eleven genes had differentially expressed mRNA levels, which were used to build the model for cancer detection. The final detection models consisting of mRNA expression levels of these nine genes had great performances on both training group and testing group. The model that constructed in this study suggested MSX1 and DCLK1 might be used in colorectal cancer detection or as target of cancer therapies. J. Cell. Physiol. 232: 1879-1884, 2017. © 2016 Wiley Periodicals, Inc.

Path-following control of wheeled planetary exploration robots moving on deformable rough terrain.

The control of planetary rovers, which are high performance mobile robots that move on deformable rough terrain, is a challenging problem. Taking lateral skid into account, this paper presents a rough terrain model and nonholonomic kinematics model for planetary rovers. An approach is proposed in which the reference path is generated according to the planned path by combining look-ahead distance and path updating distance on the basis of the carrot following method. A path-following strategy for wheeled planetary exploration robots incorporating slip compensation is designed. Simulation results of a four-wheeled robot on deformable rough terrain verify that it can be controlled to follow a planned path with good precision, despite the fact that the wheels will obviously skid and slip.

[Effects of beta3-adrenergic receptor antagonist on myocardial UCP2 expression and energy metabolism in chronic heart failure rats].

OBJECTIVE: To observe the effects of beta3-adrenergic receptor(beta3-AR) antagonist on myocardial uncoupling protein 2 (UCP2) expression and energy metabolism in chronic heart failure rats. METHODS: Seven weight-matched normal adult rats (control group), 18 isoproterenol (ISO) induced heat failure (HR) rats (ISO group) and 21 ISO induced heart failure rats but received specific beta3-AR inhibitor SR59230A (ISO+ SR59230A group) for 6 weeks were included in this research. At the end of the study, echocardiography was performed, the ratio of left ventricular weight and body weight (LVW/BW) was calculated. The expression of beta3-AR ad UCP2 mRNA in myocardium were detected by reverse transcription-polymerase chain reaction (RT-PCR), the UCP2 protein in myocardium were detected by Western blot. The myocardial contents of creatine phosphate (PCr) and adenosine triphosphate (ATP) were measured by high performance liquid chromatography (HPLC). RESULTS: Compared with control group, the cardiac function was significantly reduced and myocardial beta3-AR mRNA significantly increased, UCP2 mRNA and protein were also significantly increased in ISO group, this change could be attenuated by the treatment with SR59230A, and the expression of myocardial UCP2 protein negatively correlated with the ratio of PCr/ATP. CONCLUSION: In the chronic stage of HF, the expression of UCP2 increases, which causes myocardial energy shortage, SR59230A improves myocardia energy efficiency and cardiac function by means of suppressing the expression of UCP2.

[Effects of glucose and insulin in the cell differentiation from bone marrow stem cells to osteoblasts].

OBJECTIVE: To observe the effects of different concentrations of glucose and insulin in cell differentiation from bone marrow stem cells (BMSC) into osteoblasts (OB) and explore the acting mechanism of glucose and insulin in bone metabolism. METHODS: The in vitro technique of cell culture was employed to separate, purify and cultivate BMSC from the tibia and femur of rats. Then the BMSCs were differentiated into osteoblasts under the inductions of different glucose concentrations (5.6, 25, 50 mmol/L) with or without the addition of insulin (0.6 microg/ml). Later the calcium nodule cell ratio of osteoblasts in alizarin red stain was observed microscopically to assay the mRNA expression of OB markers-ALP, BGP and Runx2 by real-time PCR. RESULTS: With the increasing glucose concentrations, the number of red calcium nodules obviously decreased. PCR showed that the mRNA expression of ALP, BGP and transcription factor Runx2 decreased too. The difference was significant (P < 0.05). The differentiation of OB decreased with with the increasing glucose concentrations. The cell number of red calcium nodules obviously increased in the same glucose concentration groups. The mRNA expression of ALP, BGP and Runx2 also increased (P < 0.01). It hinted that insulin could enhance the differentiation of OB. CONCLUSION: Glucose can inhibit the differentiation from BMSC into OB. The inhibition is more sensitive along with the concentration increments. This may be one of the pathogenesises of diabetic osteoporosis. Insulin can enhance the differentiation from BMSC into OB. In addition, it can improve the inhibition of differentiation from BMSC into OB because of a high concentration of glucose.