Overexpression of DCF1 Inhibits Glioma through Destruction of Mitochondria and Activation of Apoptosis Pathway

Gliomas are the most common brain tumors affecting the central nervous system and are associated with a high mortality rate. DCF1 is a membrane protein that was previously found to play a role in neural stem cell differentiation. In the present study, we found that overexpression of dcf1 significantly inhibited cell proliferation, migration, and invasion and dramatically promoted apoptosis in the glioblastoma U251 cell line. DCF1 deletion mutations in the functional region showed that the complete structure of DCF1 was necessary for apoptosis. Furthermore, significantly lower tumorigenicity was observed in athymic nude mice by transplanting U251 cells overexpressing dcf1. To decode the apoptosis induced by dcf1, mitochondrial structure and membrane potential in glioma cells were investigated and the results indicated obvious mitochondrial swelling, destruction of cristae, and a significant decline in membrane potential. Mechanismly, caspase-3 signaling was activated. Finally, endogenous dcf1 silence in U251 cells was investigated. Results showed a highly methylation at -1339 and -1322 position at dcf1 promoter sequence, revealing the causal relationship between dcf1 gene and tumorigencicity. The present study identified a previously unknown cancer apoptosis mechanism involving dcf1 overexpression and provided a novel approach to potentially treat glioma patients

Antimikrobna rezistencija i svojstva virulencije bakterije Enterococcus faecium izolirane u goveda s kliničkim mastitisom iz pokrajine Ningxia, Kina

This study was conducted to determine the antimicrobial resistance and virulence traits of 32 Enterococcus faecium isolates from clinical bovine mastitis cases in Ningxia Province, China. In total, 32 E. faecium isolates were taken from subclinical bovine mastitis on the basis of morphological characterization and biochemical testing, and screened for antimicrobial susceptibility. The virulence genes of the isolates were studied using polymerase chain reaction (PCR). The disc diffusion assay revealed a high occurrence of resistance against tetracycline (78.1%) and erythromycin (68.8%) in the E. faecium isolates. However, all tested E. faecium were susceptible to linezolid and vancomycin. Moreover, all E. faecium isolates harbored the erythromycin-resistant genes ermA, ermB and ermC, as well as the tetracycline-resistant genes tetK, tetL and tetM. Furthermore, all E. faecium isolates carried more than 3 of the tested virulence genes. The presence of agg (100%), cpd (100%), efaA (100%), gelE (93.4%), and esp (75.0%) was found most frequently in all the tested isolates. These findings are useful for making appropriate antimicrobial choices and developing antivirulence therapies for subclinical bovine mastitis caused by E. faecium in Ningxia Province, China.Istraživanje je provedeno kako bi se odredila antimikrobina rezistencija i svojstva virulencije izolata bakterije Enterococcus faecium uzetih u goveda s kliničkim mastitisom. U ukupno 32 izolata goveda iz pokrajine Ningxia u Kini, procijenjena je antimikrobna osjetljivost na temelju morfološke karakterizacije i biokemijskih pretraga. Geni virulencije izolata istraženi su polimeraznom lančanom reakcijom (PCR). Disk-difuzijski test je u izolatu bakterije E. faecium pokazao visoku pojavnost rezistencije na tetraciklin (78,1 %) i eritromicin (68,8 %). Svi su pretraženi izolati bili osjetljivi na linezolid i vankomicin i imali gene rezisentne na eritromicin ermA, ermB i ermC, kao i na tetraciklin, tetK, tetL i tetM. Osim toga svi izolati E. faecium nosili su više od tri istraživana gena virulencije. Najčešći geni bili agg (100 %), cpd (100 %), efaA (100 %), gelE (93,4 %) i esp (75,0 %). Ovi rezultati mogu u pokrajini Ningxia u Kini pridonijeti pravilnom izboru antimikrobnog lijeka i razvoju uspješne terapije za supklinički goveđi mastitis uzrokovan bakterijom E. faecium

Antisense inhibition of ATM gene enhances the radiosensitivity of head and neck squamous cell carcinoma in mice

<p>Abstract</p> <p>Background</p> <p>Treatment failure after radiotherapy of head and neck squamous cell carcinoma (HNSCC) could be a significant problem. Our objective is to sensitize SCCVII cells to ionizing radiation <it>in vitro </it>and <it>in vivo </it>through inhibiting ATM expression using antisense oligodeoxynucleotides (AS-ODNs), and investigate the potential mechanism of radiosensitization.</p> <p>Methods</p> <p>We designed and synthesized AS-ODNs that target ATM mRNA to reduce the ATM expression. The influence on the expression of ATM mRNA and protein in SCCVII cells were analysed by real-time quantitative PCR and western blotting respectively. Clonogenic survival assay was performed to detect the survival ability of SCCVII cells after irradiation, while flow cytometry used to analyse the cell cycle and apoptosis. The volume of solid tumors generated with SCCVII cells was measured, and cell apoptosis was analysed by TUNEL assay after irradiation.</p> <p>Results</p> <p>The relative ATM mRNA and protein expression in SCCVII cells treated with ATM AS-ODNs were decreased to 25.7 ± 3.1% and 24.1 ± 2.8% of that in untreated cells respectively (<it>P </it>< 0.05). After irradiation, the survival fraction (SF) of cells treated with ATM AS-ODNs was lower than that of other groups at the same dose of radiation (<it>P </it>< 0.05), while the percentage of cells in G2/M phase decreased and apoptotic rate of cells increased(<it>P </it>< 0.05). The inhibition rate in SCCVII cells solid tumor exposed to X-ray alone was 23.2 ± 2.7%, while it was 56.1 ± 3.8% in the group which irradiated in combination with the treatment of ATM AS-ODNs (<it>P </it>< 0.05). The apoptotic index for the group irradiated in combination with ATM AS-ODNs injection was 19.6 ± 3.2, which was significantly higher than that of others (<it>P </it>< 0.05)</p> <p>Conclusion</p> <p>Inhibition of ATM expression sensitized SCCVII cells to ionizing radiation <it>in vitro </it>and <it>in vivo</it>. The potential mechanism should be the defective G2/M cell cycle checkpoint control and enhanced radiation-induced apoptosis.</p

比利时

peer reviewedQuinoa is an excellent source of nutritional and bioactive components. Protein is considered a key nutritional advantage of quinoa grain, and many studies have highlighted the nutritional and physicochemical properties of quinoa protein. In addition, quinoa protein is a good precursor of bioactive peptides. This review focused on the biological properties of quinoa protein hydrolysate and peptides, and gave a summary of the preparation and functional test of quinoa protein hydrolysate and peptides. A combination of milling fractionation and solvent extraction is recommended for the efficient production of quinoa protein. The biological functionalities of quinoa protein hydrolysate, including antidiabetic, antihypertensive, anti-inflammatory, antioxidant activities, and so on, have been extensively investigated based on in vitro studies and limited animal models. Additionally, bioinformatics analysis, including proteolysis simulation, virtual screening, and molecular docking, provides an alternative or assistive approach for exploring the potential bioactivity of quinoa protein and peptides. Nevertheless, further research is required for industrial production of bioactive quinoa peptides, verification of health benefits in humans, and mechanism interpretation of observed effects

Preparation, purification, and identification of novel antioxidant peptides derived from Gracilariopsis lemaneiformis protein hydrolysates

Gracilariopsis lemaneiformis (G. lemaneiformis) protein was hydrolyzed with alkaline protease to obtain antioxidant peptides. The enzymatic hydrolysis conditions were optimized through single-factor and orthogonal experiments. The results showed that the optimal process parameters were using 2% of alkaline protease, and substrate concentration of 1 g/100 mL and hydrolyzed 2 h at pH 8.0. Gel filtration chromatography and RP-HPLC were adopted for isolating and purifying the antioxidant peptides from the G. lemaneiformis protein hydrolysate (GLPH). Three novel antioxidant peptides were identified as LSPGEL (614.68 Da), VYFDR (698.76 Da), and PGPTY (533.57 Da) by nano-HPLC-MS/MS. The results of ABTS free radical scavenging rate demonstrated PGPTY exhibited the best antioxidant activity (IC50 = 0.24 mg/mL). Moreover, LSPGEL, VYFDR, and PGPTY were docked with Keap1, respectively. The molecular docking results suggested PGPTY had smaller docking energy and inhibition constants than the other two peptides. Finally, the cell viability assay evidenced the protective effect exerted by the antioxidant peptide on H2O2-induced oxidative damage. Above findings showed the potential of using antioxidant peptides from GLPH as antioxidants

Exposure to low-level metalaxyl impacts the cardiac development and function of zebrafish embryos.

Metalaxyl is an anilide pesticide that is widely used to control plant diseases caused by Peronosporales species. In order to study the toxic effects, zebrafish embryos were exposed to metalaxyl at nominal concentrations of 5, 50 and 500 ng/L for 72 hr, and the cardiac development and functioning of larvae were observed. The results showed that metalaxyl exposure resulted in increased rates of pericardial edema, heart hemorrhage and cardiac malformation. The distance between the sinus venosus and bulbus arteriosus, stroke volume, cardiac output and heart rate were significantly increased in larvae exposed to 50 and 500 ng/L metalaxyl compared to solvent control larvae. Significant upregulation in the transcription of tbx5, gata4 and myh6 was observed in the 50 and 500 ng/L treatments, and that of nkx2.5 and myl7 was observed in the 5, 50 and 500 ng/L groups. These disturbances may be related to cardiac developmental and functional defects in the larvae. The activity of Na+/K+-ATPase and Ca2+-ATPase was significantly increased in zebrafish embryos exposed to 500 ng/L metalaxyl, and the mRNA levels of genes related to ATPase (atp2a11, atp1b2b, and atp1a3b) (in the 50 and 500 ng/L groups) and calcium channels (cacna1ab) (in the 500 ng/L group) were significantly downregulated; these changes might be associated with heart arrhythmia and functional failure

Comprehensive Evaluation of Tea Cultivars Suitable for Matcha Production Using Multivariate Statistical Analysis

Matcha was prepared from 36 tea cultivars grown in the same tea garden according to the shading requirements for fresh leaves to be used for the production of matcha and its 11 quality indexes such as sensory quality, major physicochemical properties and chroma values were analyzed. In order to select tea cultivars suitable for the manufacturing of matcha, a comprehensive evaluation model of matcha quality was established by cluster analysis (CA), principal component analysis (PCA) and multiple linear regression analysis. The CA results showed that the 36 cultivars could be divided into three groups. Matcha from group I had the best quality with green color, fresh and mellow taste, and low phenol/ammonia ratio. Matcha from group II had high phenol/ammonia ratio and strong astringent taste. Matcha from Group III, consisting of etiolated and albino cultivars, had poor color and aroma quality. The PCA results showed that the cumulative contribution rate of the first five principal components was 88.152%. Comprehensive evaluation of matcha using the evaluation function constructed based on the first five principal components showed that the top 10 cultivars were Zhongcha 102, Taicha 12, Zhongcha 108, Fuding Dahao, Meizhan, Fuding Dabai, Fuyun 6, Zi Mudan, Maolv and Yingshuang. The model describing the relationship between sensory quality and physicochemical properties established by multiple linear regression analysis was as follows: y = 3.167|a*| + 46.850 (R2 = 0.710, P < 0.001). The scores of matcha cultivars evaluated by this model were highly consistent with the comprehensive evaluation results based on principal components, indicating that the a* value of dried tea could be used as a representative index to evaluate the quality of matcha. The results of this study can provide a reference for evaluating the suitability of tea cultivars for matcha manufacturing

Identification of antibodies with non-overlapping neutralization sites that target coxsackievirus A16

手足口病（Hand, Foot and Mouth Disease，HFMD）是一种由人肠道病毒引起的全球性传染病，主要发生于5岁以下的婴幼儿。2月5日，我校夏宁邵教授团队在《细胞》子刊《细胞•宿主与微生物》（Cell Host & Microbe）上在线发表题为“Identification of antibodies with non-overlapping neutralization sites that target coxsackievirus A16”的研究论文。该研究首次揭示了手足口病主要病原体柯萨奇病毒A组16型（CVA16）三种衣壳颗粒形式与三种不同类型的治疗性中和抗体的全面相互作用细节和非重叠的中和表位结构信息，阐明了CVA16成熟颗粒是疫苗候选主要保护性免疫原的理论基础，建立了可指导疫苗研制的免疫原特异检测方法，为CVA16疫苗及抗病毒药物研究提供关键基础。我校夏宁邵教授、李少伟教授、程通副教授和美国加州大学洛杉矶分校纳米系统研究所Z. Hong Zhou（周正洪）教授为该论文的共同通讯作者。我校博士生何茂洲、徐龙发博士后、郑清炳高级工程师、博士生朱瑞和尹志超为该论文共同第一作者。【Abstract】Hand, foot, and mouth disease is a common childhood illness primarily caused by coxsackievirus A16 (CVA16), for which there are no current vaccines or treatments. We identify three CVA16-specific neutralizing monoclonal antibodies (nAbs) with therapeutic potential: 18A7, 14B10, and NA9D7. We present atomic structures of these nAbs bound to all three viral particle forms—the mature virion, A-particle, and empty particle—and show that each Fab can simultaneously occupy the mature virion. Additionally, 14B10 or NA9D7 provide 100% protection against lethal CVA16 infection in a neonatal mouse model. 18A7 binds to a non-conserved epitope present in all three particles, whereas 14B10 and NA9D7 recognize broad protective epitopes but only bind the mature virion. NA9D7 targets an immunodominant site, which may overlap the receptor-binding site. These findings indicate that CVA16 vaccines should be based on mature virions and that these antibodies could be used to discriminate optimal virion-based immunogens.This work was supported by grants from the Major Program of National Natural Science Foundation of China ( 81991490 ), the National Science and Technology Major Projects for Major New Drugs Innovation and Development ( 2018ZX09711003-005-003 ), the National Science and Technology Major Project of Infectious Diseases ( 2017ZX10304402-002-003 ), the National Natural Science Foundation of China ( 31670933 and 81801646 ), the China Postdoctoral Science Foundation ( 2018M640599 and 2019T120557 ), the Principal Foundation of Xiamen University ( 20720190117 ), and the National Institutes of Health ( R37-GM33050 , GM071940 , DE025567 , and AI094386 ). 该研究获得了国家自然科学基金、新药创制国家科技重大专项、传染病防治国家科技重大专项和美国国立卫生研究院基金的资助