ABSTRACT
Development of the endosperm is strikingly different in monocots and dicots: it often manifests as a persistent tissue in the former and transient tissue in the latter. Little is known about the controlling mechanisms responsible for these different outcomes. Here we characterized a maize (Zea mays) mutant, endosperm breakdown1 (enb1), in which the typically persistent endosperm (PE) was drastically degraded during kernel development. ENB1 encodes a cellulose synthase 5 that is predominantly expressed in the basal endosperm transfer layer (BETL) of endosperm cells. Loss of ENB1 function caused a drastic reduction in formation of flange cell wall ingrowths (ingrowths) in BETL cells. Defective ingrowths impair nutrient uptake, leading to premature utilization of endosperm starch to nourish the embryo. Similarly, developing wild-type kernels cultured in vitro with a low level of sucrose manifested early endosperm breakdown. ENB1 expression is induced by sucrose via the BETL-specific Myb-Related Protein1 transcription factor. Overexpression of ENB1 enhanced development of flange ingrowths, facilitating sucrose transport into BETL cells and increasing kernel weight. The results demonstrated that ENB1 enhances sucrose supply to the endosperm and contributes to a PE in the kernel.
Subject(s)
Endosperm , Zea mays , Cell Wall/metabolism , Endosperm/metabolism , Glucosyltransferases , Sucrose/metabolism , Zea mays/metabolismABSTRACT
Strain effect in the structurally defective materials can contribute to the catalysis optimization. However, it is challenging to achieve the performance improvement by strain modulation with the help of geometrical structure because strain is spatially dependent. Here, a new class of compressively strained platinum-iridium-metal zigzag-like nanowires (PtIrM ZNWs, M = nickel (Ni), cobalt (Co), iron (Fe), zinc (Zn) and gallium (Ga)) is reported as the efficient alkaline hydrogen evolution reaction (HER) and hydrogen oxidation reaction (HOR) catalysts. Particularly, the optimized PtIrNi ZNWs with 3% compressive strain (cs-PtIrNi ZNWs) can achieve the highest HER/HOR performances among all the catalysts investigate. Their HOR mass and specific activities are 3.2/14.4 and 2.6/32.7 times larger than those of PtIrNi NWs and commercial Pt/C, respectively. Simultaneously, they can exhibit the superior stability and high CO resistance for HOR. Further, experimental and theoretical studies collectively reveal that the compressive strain in cs-PtIrNi ZNWs effectively weakens the adsorption of hydroxyl intermediate and modulates the electronic structure, resulting in the weakened hydrogen binding energy (HBE) and moderate hydroxide binding energy (OHBE), beneficial for the improvement of HOR performance. This work highlights the importance of strain tuning in enhancing Pt-based nanomaterials for hydrogen catalysis and beyond.
ABSTRACT
The electroreduction of carbon dioxide into high-value-added products is an effective approach to alleviating the energy crisis and pollution issues. However, there are still significant challenges for multicarbon (C2+) product production due to the lack of efficient catalysts with high selectivity. Herein, a Cu-rich electrocatalyst, where Cu2O nanoparticles are decorated on two-dimensional (2D) Cu-BDC metal-organic frameworks (MOFs) with abundant heterogeneous interfaces, is synthesized for highly selective CO2 electroreduction into C2+ products. A high C2+ Faradaic efficiency of 72.1% in an H-type cell and 58.2% in a flow cell are obtained, respectively. The heterogeneous interfaces of Cu2O/Cu-BDC can optimize the adsorption energy of reaction intermediates during CO2 electroreduction. An in situ infrared spectroscopy study indicates that the constructed interfaces can maintain the particular distribution of Cu valence states, where the C-C coupling is promoted to efficiently produce C2+ products owing to the stabilization of *CHO and *COH intermediates.
ABSTRACT
Adult zebrafish is an emerging vertebrate model for studying genetic basis of cardiomyopathies; but whether the simple fish heart can model essential features of hypertrophic cardiomyopathy (HCM) remained unknown. Here, we report a comprehensive phenotyping of a lamp2 knockout (KO) mutant. LAMP2 encodes a lysosomal protein and is a causative gene of Danon disease that is characterized by HCM and massive autophagic vacuoles accumulation in the tissues. There is no effective therapy yet to treat this most lethal cardiomyopathy in the young. First, we did find the autophagic vacuoles accumulation in cardiac tissues from lamp2 KO. Next, through employing a set of emerging phenotyping tools, we revealed heart failure phenotypes in the lamp2 KO mutants, including decreased ventricular ejection fraction, reduced physical exercise capacity, blunted ß-adrenergic contractile response, and enlarged atrium. We also noted changes of the following indices suggesting cardiac hypertrophic remodeling in lamp2 KO: a rounded heart shape, increased end-systolic ventricular volume and density of ventricular myocardium, elevated actomyosin activation kinetics together with increased maximal isometric tension at the level of cardiac myofibrils. Lastly, we assessed the function of lysosomal-localized mTOR on the lamp2-associated Danon disease. We found that haploinsufficiency of mtor was able to normalize some characteristics of the lamp2 KO, including ejection fraction, ß-adrenergic response, and the actomyosin activation kinetics. In summary, we demonstrate the feasibility of modeling the inherited HCM in the adult zebrafish, which can be used to develop potential therapies.
Subject(s)
Glycogen Storage Disease Type IIb/metabolism , Lysosomal-Associated Membrane Protein 2/genetics , Phenotype , TOR Serine-Threonine Kinases/antagonists & inhibitors , Zebrafish/genetics , Animals , Cardiomegaly/genetics , Disease Models, Animal , Gene Knockout Techniques , Glycogen Storage Disease Type IIb/genetics , Lysosomal-Associated Membrane Protein 2/metabolism , Myocardial Contraction/genetics , Myocardium/metabolism , Myofibrils/metabolism , Receptors, Adrenergic, beta/metabolism , Stroke Volume , TOR Serine-Threonine Kinases/genetics , TOR Serine-Threonine Kinases/metabolism , Ventricular Remodeling/genetics , Zebrafish/metabolismABSTRACT
To investigate the effect of microRNA 21 (miR-21) on hepatic stellate cells (HSCs) proliferation and apoptosis, and further to study its potential mechanisms. LX-2 cells were divided into miR-21 mimic group (Mimic), miR-21 mimic negative control group (NM), miR-21 inhibitor group (Inhibitor), miR-21 inhibitor negative control group (NC), and blank control group (Control). The cell proliferation was detected by CCK-8 assay and the cell migration and invasion were detected by scratch and transwell assay. Cell cycle and apoptosis were detected by flow cytometry. The levels of interleukin (IL)-6, tumor necrosis factor (TNF)-α, and transforming growth factor (TGF)-ß1 were detected by enzyme-linked immunosorbent assay (ELISA). Proliferation, apoptosis, and phosphatase and tensin homolog (PTEN)/phosphoinositide 3-kinase (PI3K)/protein kinase B (AKT) signaling pathway related genes and proteins were detected by quantitative real-time polymerase chain reaction (qRT-PCR) and western blot, respectively. The cells proliferation, migration, and invasion were promoted in Mimic group. The levels of IL-6, TNF-α, and TGF-ß1 were increased after miR-21 administration. The expression of α-smooth muscle actin (SMA) and collagen 1 (Colla1) were increased, while Bax/B-cell lymphoma (Bcl)-2 ratio and programed cell death 4 (PDCD4) were reduced after miR21 treatment. Meanwhile, the mRNA and protein expression of PTEN were reduced and PI3K/AKT pathway been promoted. Our study demonstrated that miR-21 could promote proliferation and inhibit apoptosis of HSCs, and its mechanism may be related to PTEN/PI3K/AKT pathway.
Subject(s)
Apoptosis/drug effects , Hepatic Stellate Cells/metabolism , MicroRNAs/genetics , PTEN Phosphohydrolase/genetics , Actins/genetics , Apoptosis Regulatory Proteins/genetics , Cell Line , Cell Movement/drug effects , Cell Proliferation/genetics , Collagen/genetics , Hepatic Stellate Cells/drug effects , Humans , Interleukin-6/genetics , MicroRNAs/pharmacology , Phosphatidylinositol 3-Kinase/genetics , Proto-Oncogene Proteins c-akt/genetics , RNA-Binding Proteins/genetics , Transforming Growth Factor beta1/genetics , Tumor Necrosis Factor-alpha/geneticsABSTRACT
Bimetallic PtRu are promising electrocatalysts for hydrogen oxidation reaction in anion exchange membrane fuel cell, where the activity and stability are still unsatisfying. Here, PtRu nanowires were implanted with a series of oxophilic metal atoms (named as i-M-PR), significantly enhancing alkaline hydrogen oxidation reaction (HOR) activity and stability. With the dual doping of In and Zn atoms, the i-ZnIn-PR/C shows mass activity of 10.2 A mgPt+Ru-1 at 50 mV, largely surpassing that of commercial Pt/C (0.27 A mgPt-1) and PtRu/C (1.24 A mgPt+Ru-1). More importantly, the peak power density and specific power density are as high as 1.84 W cm-2 and 18.4 W mgPt+Ru-1 with a low loading (0.1 mg cm-2) anion exchange membrane fuel cell. Advanced experimental characterizations and theoretical calculations collectively suggest that dual doping with In and Zn atoms optimizes the binding strengths of intermediates and promotes CO oxidation, enhancing the HOR performances. This work deepens the understanding of developing novel alloy catalysts, which will attract immediate interest in materials, chemistry, energy and beyond.
ABSTRACT
BACKGROUND: The Roche cobas TaqScreen MPX test was used to evaluate the rate of hepatitis B surface antigen (HBsAg)-negative donations that were hepatitis B virus (HBV) DNA reactive from June 2010 to January 2011 in Qingdao, China. STUDY DESIGN AND METHODS: HBsAg-negative samples from 65,800 voluntary blood donors were tested with the cobas TaqScreen MPX test in pools of 6 on the Roche cobas s 201 blood screening platform. Samples positive for HBV DNA and negative for HBsAg were quantitated with the Roche COBAS AmpliPrep/COBAS TaqMan HBV test. In addition, serologic tests for HBsAg, hepatitis B surface antibody, anti-hepatitis B core antigen (anti-HBc), anti-hepatitis B e antigen (anti-HBe), and hepatitis B e antigen (HBe) were done using the Roche electrochemiluminescence immunoassay. RESULTS: A total of 80 nucleic acid amplification technology (NAT) test-reactive pools were identified and 59 pools (74%) resolved to a reactive sample. All samples were HBV DNA reactive and the viral load in each sample was quantitated. The viral loads of the samples ranged from less than 20 to 34,600 IU/mL; 13 samples (22%) had viral loads of more than 20 IU/mL, 27 samples (45.8%) had viral loads of less than 20 IU/mL, and 19 samples (32.2%) had undetectable viral loads. Of the 59 NAT-reactive samples, 40 (67.8%) were anti-HBc positive. Fifteen of the 59 samples could not be confirmed as NAT reactive either by an alternative NAT test or by serology. CONCLUSION: The HBV NAT yield in blood donors in Qingdao is 0.06% (38/65,800). This study confirmed the value of NAT for interdicting HBV-positive donations and preventing transfusion-transmitted HBV infections.
Subject(s)
Blood Donors , HIV-1/isolation & purification , HIV-2/isolation & purification , Hepacivirus/isolation & purification , Hepatitis B virus/isolation & purification , Multiplex Polymerase Chain Reaction/methods , Real-Time Polymerase Chain Reaction/methods , Adult , Blood Banks , China , DNA, Viral/genetics , DNA, Viral/isolation & purification , HIV Antibodies/analysis , HIV Antibodies/blood , HIV-1/genetics , HIV-2/genetics , Hepacivirus/genetics , Hepatitis Antibodies/analysis , Hepatitis Antibodies/blood , Hepatitis B virus/genetics , Humans , Mass Screening/methods , Mass Screening/statistics & numerical data , Middle Aged , Serologic Tests/methods , Serologic Tests/statistics & numerical data , Young AdultABSTRACT
Modifier genes contribute significantly to our understanding of pathophysiology in human diseases; however, effective approaches to identify modifier genes are still lacking. Here, we aim to develop a rapid F0-based genetic assay in adult zebrafish using the bag3 gene knockout (bag3e2/e2) cardiomyopathy model as a paradigm. First, by utilizing a classic genetic breeding approach, we identified dnajb6b as a deleterious modifier gene for bag3 cardiomyopathy. Next, we established an F0-based genetic assay in adult zebrafish through injection of predicted microhomology-mediated end joining (MMEJ)-inducing single guide RNA/Cas9 protein complex. We showed that effective gene knockdown is maintained in F0 adult fish, enabling recapitulation of both salutary modifying effects of the mtor haploinsufficiency and deleterious modifying effects of the dnajb6b gene on bag3 cardiomyopathy. We finally deployed the F0-based genetic assay to screen differentially expressed genes in the bag3 cardiomyopathy model. As a result, myh9b was identified as a novel modifier gene for bag3 cardiomyopathy. Together, these data prove the feasibility of an F0 adult zebrafish-based genetic assay that can be effectively used to discover modifier genes for inherited cardiomyopathy.
Subject(s)
Cardiomyopathies , Zebrafish , Adaptor Proteins, Signal Transducing/metabolism , Animals , Apoptosis Regulatory Proteins/genetics , Cardiomyopathies/genetics , Gene Knockout Techniques , Genes, Modifier , Zebrafish/genetics , Zebrafish/metabolism , Zebrafish Proteins/genetics , Zebrafish Proteins/metabolism , RNA, Guide, CRISPR-Cas SystemsABSTRACT
Herein, we demonstrate that the vulcanization of copper nanosheets (Cu2S NSs) can realize selective electrochemical carbon dioxide reduction (CO2RR) to formate (HCOOH). Results from experiments and in situ infrared spectroscopy reveal that Cu2S NSs favor CO2 reduction along the HCOO* pathway, leading to the selective formation of HCOOH. Impressively, the Faraday efficiency (FE) of HCOOH surpasses 70% in a broad potential range from -0.84 to -1.21 V versus (vs.) reversible hydrogen electrode (RHE), with a maximum value of 82%.
ABSTRACT
Bisphenol analogues are widely used in industrial and daily-use consumer products having imperfect thyroid hormones (THs) structures. Widespread exposure interferes with thyroid-related health outcomes in human. The mechanisms of disruption on TH synthesis and subsequent thyroid dysfunction by different bisphenol analogues remain unclear. Here, we evaluated bisphenol-induced thyroid endocrine disruption in C57BL/6 mice at doses of 0.002, 0.02, 2, and 20 mg/kg body weight/day (BW/d) for five consecutive weeks. Administration of 20 mg/kg BW/d bisphenol S (BPS) and 2 mg/kg BW/d tetrabromobisphenol S (TBBPS) significantly increased serum thyrotropin (TSH) levels to 1.21-fold and 1.20-fold of control group, respectively, indicating that bisphenols induced thyroid dysfunction in mice. Height of the thyroid follicle epithelium significantly increased to 1.27-, 1.24-, 1.26-, and 1.36-fold compared to control group with BPA, BPS, TBBPA, and TBBPS at 20 mg/kg BW/d, respectively, indicating impairment of the thyroid gland structure, and TBBPS showed potent effect. Exposure to bisphenol analogues of 0.02 mg/kg BW/d downregulated the protein expression levels of thyrotropin receptor, the sodium/iodide symporter, thyroperoxidase. The TH-dependent effects were further determined using the T-Screen assay at 10-11 M to 10-5 M concentrations. Bisphenol analogues significantly decreased TH-dependent GH3 cell proliferation, indicating the antagonistic activity of bisphenol analogues. The gene responsible for THs synthesis of thyrotropin releasing hormone receptor and TSH were upregulated, but downregulation of thyroid receptor ß was observed. Our results suggest that bisphenol analogues distinctly induce thyroid dysfunction via TH synthesis, implying adverse effect of bisphenol analogues on TH homeostasis and subsequent physiological processes.
Subject(s)
Thyroid Gland , Thyroid Hormones , Mice , Humans , Animals , Thyroid Gland/metabolism , Mice, Inbred C57BL , Thyroid Hormones/metabolism , Benzhydryl Compounds/toxicity , ThyrotropinABSTRACT
Transverse aortic constriction (TAC) is a frequently used model to investigate pressure overload-induced progressive heart failure (HF); however, there is considerable phenotypic variation among different mouse strains and even sub-strains. Moreover, less is known about the TAC model in ICR mice. Therefore, to determine the suitability of the ICR strain for TAC-induced HF research, we compared the effects of TAC on ICR and C57BL/6J mice at one, two and four weeks post-TAC via echocardiography, organ index, morphology, and histology. At the end of the study, behavior and gene expression patterns were assessed, and overall survival was monitored. Compared to the sham-operated mice, ICR and C57BL/6J mice displayed hypertrophic phenotypes with a significant increase in ventricle wall thickness, heart weight and ratio, and cross-sectional area of cardiomyocytes after a 2-week TAC exposure. In addition, ICR mice developed reduced systolic function and severe lung congestion 4 weeks post-TAC, whereas C57BL/6J did not. Besides, ICR mice demonstrated comparable survival, similar gene expression alteration but severer fibrotic remodeling and poor behavioral performance compared to the C57BL/6J mice. Our data demonstrated that ICR was quite sensitive to TAC-induced heart failure and can be an ideal research tool to investigate mechanisms and drug intervention for pressure overload-induced HF.
ABSTRACT
BACKGROUND: The university environment poses a high risk of spreading infectious diseases, particularly the 2009 pandemic influenza H1N1, as it is a mass gathering place for youth. This study aimed to evaluate the predictors of stress symptoms among Chinese university students during the 2009 H1N1 influenza pandemic. MATERIAL/METHODS: We used a self-reported questionnaire, the PTSD (posttraumatic stress disorder) Checklist-Civilian Version (PCL-C) to evaluate the stress symptoms among Chinese university students from Heilongjiang (n=455), Beijing (n=106), Shanghai (n=419) and Sichuan (n=102). We then analyzed the predictors of stress symptoms. RESULTS: The proportion of university students enrolled in this study who met symptomatic criteria for PTSD was 2% (22 students). The mean PCL-C total score in the sample was 22.09 ± 8.01. The correlational analyses revealed a significant positive relationship between the PCL-C total score and area, and university grade (P<0.01). Moreover, a negative relationship was found between the PCL-C total score and gender, having H1N1 influenza, having family members, friends or acquaintances having H1N1 influenza, and being afraid of H1N1 influenza (P<0.01). The regression analyses showed that in North China, female gender, having H1N1 influenza, having family members or acquaintances with H1N1 influenza, and being afraid of H1N1 influenza were significant predictors of the stress symptoms. CONCLUSIONS: In North China, female gender, having H1N1 influenza, having family members, friends, or acquaintances with H1N1 influenza, and being afraid of H1N1 influenza were significant predictors of the stress symptoms.
Subject(s)
Influenza A Virus, H1N1 Subtype , Influenza, Human/complications , Risk Factors , Stress Disorders, Post-Traumatic/epidemiology , Stress Disorders, Post-Traumatic/etiology , China/epidemiology , Female , Humans , Male , Regression Analysis , Sex Factors , Surveys and Questionnaires , Young AdultABSTRACT
Osteoarthritis (OA) is a joint disease characterised by progressive cartilage degradation and inflammation, but the detailed pathogenesis of OA remains unclear. The present study aimed to investigate the role of long intergenic non-coding RNA (lincRNA)-Cox2 in OA progression and the potential mechanism. An OA mouse model was used for in vivo experiments, and IL-1ß-induced injury of mouse chondrocytes was conducted for in vitro experiments. Small interfering (si)-Cox2 was transfected into chondrocytes to elucidate the effect of lincRNA-Cox2 on OA. Quantitative reverse transcription PCR assays were conducted to detect the expression of lincRNA-Cox2 and microRNA (miR)-150. Cell proliferation and apoptosis were analysed based on an MTT assay and annexin V/propidium iodide staining, respectively. Western blotting was performed to evaluate the protein expression levels of Ki-67, PCNA, Bax, cleaved (c)-Caspase-3, c-Caspase-9 and Wnt/ß-catenin pathway-associated proteins in chondrocytes. High levels of lincRNA-Cox2 were observed in cartilage tissues of the OA mouse model in vivo. In the in vitro experiments, the expression of lincRNA-Cox2 was increased in IL-1ß-treated chondrocytes. Knockdown of lincRNA-Cox2 promoted the proliferation and inhibited the apoptosis of chondrocytes. Mechanistically, lincRNA-Cox2 was found to directly target miR-150, acting as a competing endogenous RNA, and the effect of si-Cox2 on the proliferation and apoptosis of chondrocytes was reversed by miR-150 inhibitors. Moreover, lincRNA-Cox2 activated the Wnt/ß-catenin pathway to regulate chondrocyte proliferation and apoptosis. The present study demonstrated that silencing lincRNA-Cox2 expression plays a protective role in OA by enhancing the proliferation and suppressing the apoptosis of chondrocytes, which is related to increased miR-150 expression and activation of the Wnt/ß-catenin pathway.
ABSTRACT
OBJECTIVE: Psychogenic non-epileptic seizures (PNES) are conversion disorders with functional neurological symptoms that can resemble epileptic seizures (ES). We conducted a systematic review to obtain an overview of the value of prolactin (PRL) levels in the differential diagnosis between PNES and ES. METHODS: We searched PubMed, EMBASE, and Cochrane Library databases for studies published up to June 4th, 2020. Published studies were included if they fulfilled the following criteria: original research on PRL changes after ES and PNES. By applying Bayes' theorem, we calculated the predicted values of PRL with pretest probabilities of 90 % and 75 % in ES. RESULTS: Sixteen studies were included in this review. All the studies showed that PRL levels increase after ES, especially 10-20 min after ES, when the elevation was most obvious. In studies where capillary PRL level measurements were included, the median sensitivity in the diagnosis of ES (all epileptic seizure types), generalized tonic clonic seizures (GTCS), focal impaired awareness seizures (FIAS), and focal aware seizures (FAS) was 67.3 %, 66.7 %, 33.9 %, and 11.1 %, respectively. The median specificity in the diagnosis of ES was 99.1 %. By using Bayes' theorem, when we used the median specificity and sensitivity for predictive value calculation, assuming a pretest probability of 90 %, a positive PRL measure was highly predictive (99 %) of all types of ES, and negative predictive values were all below 30 %. When we used the lowest specificity and sensitivity for predictive value calculation, assuming a pretest probability of 75 %, ES and GTCS had positive predictive values of 77.2 % and 81.0 %, respectively; the negative predictive values of PRL in ES and GTCS were 26.2 % and 29.6 %, respectively. CONCLUSIONS: The use of PRL could be a useful adjunct to differentiate GTCS from PNES. However, PRL levels are of limited use for differentiating FIAS or FAS from PNES, and a negative PRL measure is not predictive of PNES.
Subject(s)
Epilepsy , Bayes Theorem , Diagnosis, Differential , Electroencephalography , Epilepsies, Partial , Epilepsy/diagnosis , Humans , Prolactin , Seizures/diagnosisABSTRACT
The Guanxin Suhe pill (GSP), a traditional Chinese medicine, has been widely used to treat angina pectoris (AP) in Chinese clinical practice. However, research on the bioactive ingredients and underlying mechanisms of GSP in AP remains scarce. In this study, a system pharmacology approach integrating gastrointestinal absorption (GA) evaluation, drug-likeness (DL) evaluation, target exploration, protein-protein-interaction analysis, Gene Ontology (GO) enrichment analysis, network construction, and molecular docking was adopted to explore its potential mechanisms. A total of 481 ingredients from five herbs were collected, and 242 were qualified based on GA and DL evaluation. Target exploration identified 107 shared targets between GSP and AP. Protein-protein interaction identified VEGFA (vascular endothelial growth factor A), TNF (tumor necrosis factor), CCL2 (C-C motif chemokine ligand 2), FN1 (fibronectin 1), MMP9 (matrix metallopeptidase 9), PTGS2 (prostaglandin-endoperoxide synthase 2), IL10 (interleukin 10), CXCL8 (C-X-C motif chemokine ligand 8), IL6 (interleukin 6), and INS (insulin) as hub targets for GSP, which were involved in the inflammatory process, ECM proteolysis, glucose metabolism, and lipid metabolism. GO enrichment identified top pathways in the biological processes, molecular functions, and cell components, explaining GSP's potential AP treatment mechanism. Positive regulation of the nitric oxide biosynthetic process and the response to hypoxia ranked highest of the biological processes; core targets that GSP can regulate in these two pathways were PTGS2 and NOS2, respectively. Molecular docking verified the interactions between the core genes in the pathway and the active ingredients. The study lays a foundation for further experimental research and clinical application.
ABSTRACT
This study aims to investigate the effects of ß-elemene on a mouse model of heart failure (HF) and to elucidate the underlying mechanisms in vitro approaches. In this study, left anterior descending (LAD)-induced HF mouse model and oxygen-glucose deprivation/recovery (OGD/R)-induced H9C2 model were leveraged to assess the therapeutic effects of ß-elemene. Histological examination, western blot and quantitative real-time PCR analysis (RT-qPCR) and immunofluorescence staining was utilized to elucidate mechanism of ß-elemene in lipid-induced inflammation. Results showed that ß-elemene improved heart function in HF mice evidenced by the increase of cardiac ejection fraction (EF) and fractional shortening (FS) values. Furthermore, ß-elemene administration rescued ventricular dilation, lipid accumulation, and inflammatory infiltration in arginal areas of mice myocardial infarction. At transcription level, ß-elemene augmented the mRNA expression of fatty acid oxidation-associated genes, such as peroxisome proliferator-activated receptor-ß (PPARß). In vitro, treatment of ß-elemene increased carnitine palmitoyltransferase 1A (CPT1A) and sirtuin 3 (SIRT3). Hallmarks of inflammation including the nuclear translocation of nuclear factor κB (NF-κB) and the degradation of inhibitory κBα (IκBα) were significantly suppressed. Consistently, we observed down-regulation of interleukin-6 (IL-6) and pro-inflammatory cytokines (such as TNFα) in ß-elemene treated H9C2 cells. Finally, molecular docking model predicted an interaction between ß-elemene and PPARß protein. Furthermore, ß-elemene increased the expression of PPARß, which was validated by antagonist of PPARß and siRNA for PPARß.
Subject(s)
Anti-Inflammatory Agents/pharmacology , Cardiotonic Agents/pharmacology , Heart Failure/metabolism , Heart Failure/prevention & control , Inflammation/metabolism , PPAR-beta/agonists , Sesquiterpenes/pharmacology , Animals , Anti-Inflammatory Agents/therapeutic use , Cardiotonic Agents/therapeutic use , Cell Line , Cell Survival/drug effects , Disease Models, Animal , Endoribonucleases/metabolism , Heart Failure/chemically induced , Heart Failure/pathology , Inflammation/chemically induced , Lipids/toxicity , Male , Mice , Mitochondria/drug effects , Molecular Docking Simulation , Multienzyme Complexes/metabolism , NF-KappaB Inhibitor alpha/metabolism , NF-kappa B/metabolism , PPAR-beta/chemistry , PPAR-beta/genetics , PPAR-beta/metabolism , Protein Serine-Threonine Kinases/metabolism , Rats , Sesquiterpenes/chemistry , Sesquiterpenes/therapeutic useABSTRACT
Myosin heavy chain 7 (MYH7) is a major causative gene for hypertrophic cardiomyopathy, but the affected signaling pathways and therapeutics remain elusive. In this research, we identified ventricle myosin heavy chain like (vmhcl) as a zebrafish homolog of human MYH7, and we generated vmhcl frameshift mutants. We noted vmhcl-based embryonic cardiac dysfunction (VEC) in the vmhcl homozygous mutants and vmhcl-based adult cardiomyopathy (VAC) phenotypes in the vmhcl heterozygous mutants. Using the VEC model, we assessed 7 known cardiomyopathy signaling pathways pharmacologically and 11 candidate genes genetically via CRISPR/Cas9 genome editing technology based on microhomology-mediated end joining (MMEJ). Both studies converged on therapeutic benefits of mTOR or mitogen-activated protein kinase (MAPK) inhibition of VEC. While mTOR inhibition rescued the enlarged nuclear size of cardiomyocytes, MAPK inhibition restored the prolonged cell shape in the VEC model. The therapeutic effects of mTOR and MAPK inhibition were later validated in the VAC model. Together, vmhcl/myh7 loss of function is sufficient to induce cardiomyopathy in zebrafish. The VEC and VAC models in zebrafish are amenable to both efficient genetic and chemical genetic tools, offering a rapid in vivo platform for discovering candidate signaling pathways of MYH7 cardiomyopathy.
Subject(s)
Cardiomyopathies/therapy , Mitogen-Activated Protein Kinases/metabolism , TOR Serine-Threonine Kinases/metabolism , Zebrafish Proteins/metabolism , Animals , Disease Models, Animal , ZebrafishABSTRACT
PURPOSE: As a conserved cellular stress response, autophagy has recently been demonstrated to be involved in the pathogenesis of several human cancers. Beclin-1 is an important autophagy gene that is abnormally expressed in a variety of human cancers. In this study, we investigated the expression of Beclin-1 in Hepatocellular Carcinoma (HCC). METHODS: A total of 83 patients with primary HCC were enrolled in this study. The expression of Beclin-1, PCNA, NET-1, Bcl-2, and Bax was measured in tissue microarray, including 83 cases of HCC and 46 adjacent non-tumor liver tissues. The association of the expression of Beclin-1 with clinicopathological features as well as PCNA, NET-1, Bcl-2, and Bax were analyzed. RESULTS: The positive rate of Beclin-1 in HCC tissues was significantly lower than that in adjacent tissues (x2 = 4.013, p=0.012). Beclin-1 expression in HCC tissues was negatively correlated with the expression of PCNA, NET-1, and anti-apoptotic protein Bcl-2, but positively correlated with pro-apoptotic protein Bax expression. Meanwhile, Beclin-1 expression was negatively correlated with HCC Edmondson grading (p=0.0058). Furthermore, Beclin-1 expression was significantly lower in HCC patients with liver cirrhosis (p=0.029) or vascular invasion (p=0.011) than those in HCC patients without cirrhosis or vascular invasion. CONCLUSION: Decreased expression of Beclin-1 was observed in HCC tissues and negatively correlated with HCC Edmondson grading, suggesting that Beclin-1 might be a valuable prognostic indicator for HCC.
Subject(s)
Beclin-1/genetics , Carcinoma, Hepatocellular/genetics , Cell Proliferation/genetics , Liver Neoplasms/genetics , Apoptosis/genetics , Autophagy/genetics , Carcinoma, Hepatocellular/pathology , Disease Progression , Female , Gene Expression Regulation, Neoplastic/genetics , Humans , Liver Neoplasms/pathology , Male , Prognosis , Proto-Oncogene Proteins c-bcl-2/genetics , Tissue Array Analysis , bcl-2-Associated X Protein/geneticsABSTRACT
It has been reported that overexpression of the CRLF2 gene is associated with poor outcomes in pediatric B cell acute lymphoblastic leukemia (B-ALL), but the incidence rates, clinical characteristics and outcomes of CRLF2 gene overexpression in pediatric T cell ALL (T-ALL) have not been systematically analyzed. In this study, CRLF2 mRNA expression levels and clinical and laboratory parameters in 63 pediatric T-ALL patients were detected at the Children's Hospital of Chongqing Medical University and Children's Hospital of Xianyang between February 2015 and June 2018. The patients were treated according to the modified St. Jude TXV ALL protocol, and early treatment responses (bone marrow smear and MRD level) and prognoses in the enrolled patients were assessed. CRLF2 overexpression was detected in 21/63 (33.33%) patients. Statistical differences were not found for clinical or laboratory parameters (including sex, age, initial WBC count, incidence mediastinal involvement, abnormal karyotype and fusion genes) between patients with high CRLF2 expression and patients with low expression of CRLF2 (P>0.05). One patient died of tumor lysis syndrome and renal failure, and the treatment response was monitored on day 19 (TP1) of remission in 62 patients. One patient quit treatment because of family decisions, and 61 patients underwent treatment response evaluation on day 46 (TP2) of remission. Significant differences were not found between patients with high CRLF2 expression and patients with low CRLF2 expression in terms of the treatment responses at TP1 or TP2 (P>0.05). Following October 2018, 12 patients among the 61 evaluable patients relapsed (relapse rate: 19.67%), 3 patients died from chemotherapy, and the treatment-related mortality (TRM) rate was 4.92%. Secondary tumors occurred in 1 patient. The 3-year prospective EFS rate was 54.1±11.2% and 77.7±6.6% for the 61 evaluable patients and 58 patients without TRM. Patients with low CRLF2 expression had longer EFS durations than patients with high CRLF2 expression (61 evaluable patients: 35.91± 2.38 months vs 23.43± 2.57 months; 58 patients without TRM: 37.86± 2.08 months vs 24.55±2.43 months, P<0.05). CRLF2 expression levels were also monitored in 13 patients at TP1 and TP2, and the MRD level did not vary with the CRLF2 expression level. Our data suggest that clinical features, laboratory findings and treatment responses in the pediatric T-ALL population do not vary based on the overexpression of CRLF2 but that CRLF2 overexpression can contribute to a high risk of relapse in pediatric T-ALL. Thus, CRLF2 expression levels should not be used as biomarkers for monitoring MRD.
Subject(s)
Antineoplastic Combined Chemotherapy Protocols/therapeutic use , Biomarkers, Tumor/metabolism , Neoplasm Recurrence, Local/pathology , Precursor T-Cell Lymphoblastic Leukemia-Lymphoma/pathology , Receptors, Cytokine/metabolism , Adolescent , Biomarkers, Tumor/genetics , Child , Child, Preschool , Female , Follow-Up Studies , Humans , Infant , Male , Neoplasm Recurrence, Local/drug therapy , Neoplasm Recurrence, Local/genetics , Neoplasm Recurrence, Local/metabolism , Precursor T-Cell Lymphoblastic Leukemia-Lymphoma/drug therapy , Precursor T-Cell Lymphoblastic Leukemia-Lymphoma/genetics , Precursor T-Cell Lymphoblastic Leukemia-Lymphoma/metabolism , Prognosis , Prospective Studies , Receptors, Cytokine/geneticsABSTRACT
Apoptotic lymphocytes can induce specific immune tolerance. This study aimed to investigate the influence of the preoperative transfusion of apoptotic lymphocytes on allograft survival after skin transplantation. In addition, we aimed to determine changes in IL-4, IL-10 and IFN-γ mRNA expression in the grafted skin. A total of 20 New Zealand white rabbits were randomly divided into two groups: lymphocyte preconditioned group (n = 10) and control group (n = 10). Rabbits in the lymphocyte preconditioned group were intravenously injected with 60Co γ-treated donor lymphocytes at seven days before the surgery. Rabbits in the control group were intravenously injected with normal lymphocytes at seven days before skin transplantation. The mRNA expression of IL-4, IL-10 and IFN-γ in the grafted skin was determined using real-time PCR. Skin allograft rejection time was 72.63 ± 2.65 days in the lymphocyte preconditioned group and 6.52 ± 0.64 days in the control group. IL-4, IL-10 and IFN-γ mRNA expression in the skin graft was 4.32 ± 0.48, 7.86 ± 0.56 and 2.63 ± 0.25 respectively in the lymphocyte preconditioned group, and 0.58 ± 0.07, 0.91 ± 0.14 and 8.68 ± 1.23 respectively in the control group. The preoperative transfusion of apoptotic lymphocytes induced immune tolerance in the grafted skin, as demonstrated by longer survival time of the grafts before rejection. This coincided with the increased mRNA expression of IL-4 and IL-10, and the decreased expression of IFN-γ.