通过GbF3’H基因单独沉默及其与GbCHI和GbDFR基因共沉默研究其在海岛棉中抗枯萎病功能

【目的】通过沉默海岛棉GbF3’H基因及共沉默GbF3’H、GbCHI和Gb DFR基因,研究其在海岛棉抗枯萎病中的作用。【方法】以海岛棉抗病材料06-146为研究对象,GhCLA1基因为阳性对照,空载体为阴性对照,构建海岛棉TRV2-Gb F3’H沉默载体,协同课题组前期构建的TRV2-CHI和TRV2-DFR载体,利用病毒诱导的基因沉默技术(Virus-induced gene silencing,VIGS)分别进行Gb F3’H基因单独沉默以及GbF3’H、GbCHI和Gb DFR这3种基因共沉默试验。通过实时荧光定量聚合酶链式反应(Quantitative real time-polymerase chain reaction,q RT-PCR)分析各处理样品中基因沉默情况;设置室内接种枯萎病菌试验测定病情指数,分析各沉默材料对枯萎病的抗性差异。【结果】q RT-PCR结果显示,海岛棉GbF3’H基因沉默后其在海岛棉根、茎和叶中的表达量比空载体对照低,Gb F3’H、GbCHI和GbDFR这3种基因共沉默后其在海岛棉根、茎和叶中的表达量均比空载体对照低。病情指数调查结果显示,野生型<空载体对照相似文献   

丁硫克百威在豇豆不同时期施用的降解代谢研究

为明确丁硫克百威在豇豆播种期、苗期、结荚期使用后的降解代谢,以及可能产生的膳食风险,分别以其在蔬菜上登记的最低剂量、最高剂量、最高剂量的1.5倍3种剂量施药,进行田间模拟残留试验。将采集的成熟豇豆通过乙腈提取、C18分散净化,经超高效液相色谱串联质谱方法检测,测定豇豆中丁硫克百威及其代谢物——克百威和3-羟基克百威的残留量。结果表明,丁硫克百威、克百威和3-羟基克百威在豇豆中的定量限均为0.01 mg·kg^-1,在0.01~1 mg·kg^-1的添加水平下,丁硫克百威、克百威和3-羟基克百威的平均回收率为72%~105%,相对标准偏差为1.4%~20.1%。丁硫克百威使用后的超标风险主要源于其代谢物——克百威和3-羟基克百威。播种期施药后的豇豆样品均无丁硫克百威及其代谢物检出;苗期以最高剂量的1.5倍施药后,第10天的样品中克百威(含3-羟基克百威)的残留值超出中国国家标准中规定的最大允许残留限量(MRL);结荚期2次或3次施药后7 d内克百威(含3-羟基克百威)的残留值超出MRL。结荚期施药时,丁硫克百威在2次施药和3次施药后的慢性膳食摄入风险和急性膳食摄入风险均较低,小于100%;但克百威(含3-羟基克百威)的急性膳食摄入风险较高,以最高剂量的1.5倍2次施药或3次施药后,克百威(含3-羟基克百威)的急性膳食摄入风险于药后7 d才降至100%以下。综上,播种期使用丁硫克百威不会导致豇豆中残留超标,可以安全使用;但苗期和结荚期使用丁硫克百威存在极高的风险,应禁止其在播种期以外的使用。     

FAM3C activates Akt to promote viability of oral squamous-cell carcinoma cells

AIM: To investigate the expression and roles of family with sequence similarity 3, member C (FAM3C) in oral squamous-cell carcinoma cells. METHODS: The mRNA and protein expression levels of FAM3C in dysplastic oral keratinocyte (DOK) and oral squamous-cell carcinoma WSU-HN6 cells were detected by RT-qPCR and Western blot. The WSU-HN6 cells were treated with siFAM3C or FAM3C antibody. After 24, 48 and 72 h, the viability of WSU-HN6 cells was measured by CCK-8 assay, and the activation of protein kinase B (Akt) was detected by Western blot. Adenovirus was used to mediate over-expression of FAM3C in the DOK cells. The DOK cell viability was measured by CCK-8 assay after adenovirus infection for 24, 48 and 72 h, and the activation of Akt was detected by Western blot. RESULTS: Compared with the DOK cells, the mRNA and protein levels of FAM3C were significantly increased in the WSU-HN6 cells (P<0.05). The viability of WSU-HN6 cells transfected with siFAM3C was significantly inhibited at 48 h and 72 h (P<0.05). siFAM3C treatment inhibited the activation of Akt (P<0.05). FAM3C antibody treatment also suppressed the viability of the WSU-HN6 cells at 48 h and 72 h and the activation of Akt (P<0.05). Over-expression of FAM3C in the DOK cells promoted the cell viability at 48 h and 72 h and activated Akt (P<0.05). CONCLUSION: FAM3C might promote oral squamous-cell carcinoma cell growth by activating Akt.     

金属3D打印技术的发展现状及制约因素

3D打印技术作为20世纪新兴制造技术,具有高柔性化,生产周期短,材料利用率高等优点,目前在多个领域均有广泛应用。本文综述了目前3D打印技术的发展现状,常用金属3D打印技术的工作原理、特点及发展制约因素等。     

A Genome-Wide Association Study Reveals Differences in the Genetic Mechanism of Control of the Two Gait Patterns of the Brazilian Mangalarga Marchador Breed

The DMRT3 gene is described as the main gene involved in the determination of gait phenotypes in horses, and the allele A of the 22999655C>A single nucleotide polymorphisms (SNP) has been reported as a causal variant of this trait. In the Mangalarga Marchador breed, which exhibits two gait patterns with well-defined characteristics, genotypes AA and CA are associated with marcha picada and genotype CC with marcha batida. In this breed, allele A of the DMRT3 gene is only related to the marcha picada gait. The objective of this study was to identify the type of control of the marcha batida gait and to investigate SNPs and genomic regions responsible for this phenotype in Mangalarga Marchador horses. Forty-eight horses belonging to the two gait groups, marcha picada with AA and CA genotypes of the 22999655C>A SNP (n = 20) and marcha batida with CC genotype (n = 28), were analyzed using the Equine SNP70 BeadChip. The genome-wide association study result shows for the first time that, in contrast to the marcha picada gait phenotype that is apparently determined by a single gene (DMRT3) in which allele A of variant g.22999655C>A controls the trait, the marcha batida gait is controlled by a larger number of genes. Because of the small number of animals used in the two groups compared, the genomic regions associated with smaller effects on the marcha batida gait could not be identified.     

Dietary α‐linolenic acid affects lipid metabolism and tissue fatty acid profile and induces apoptosis in intraperitoneal adipose tissue of juvenile grass carp (Ctenopharyngodon idella)

A 56‐day feeding trial was conducted to elucidate the effects and mechanism action of dietary α‐linolenic acid (ALA, 18:3n‐3) on lipid accumulation and fatty acid profile of muscle, hepatopancreas and intraperitoneal fat (IPF) in juvenile grass carp using three isonitrogenous and isoenergetic semi‐purified diets containing 0.0% (control group), 1.0% and 2.0% ALA, respectively. The lowest intraperitoneal fat (IPF) ratio was found in 2.0% group. In the muscle, hepatopancreas and IPF, docosahexaenoic acid (DHA, 22:6n‐3) and eicosapentaenoic acid (EPA, 20:5n‐3) contents increased with the increase in dietary ALA. In the IPF, caspase 3, caspase 8 and caspase 9 showed the highest activities in 2.0% group, while the value of Bcl‐2/Bax (B‐cell leukaemia 2/Bcl‐2‐associated X protein) reached the lowest. Meanwhile, swelling of the IPF mitochondria was observed in 2.0% group. The gene expressions of fatty acid desaturase (FAD) and fatty acid elongase (ELO) in the hepatopancreas and muscle showed significantly higher levels in the treatment groups, whereas an opposite trend was existed in the IPF. Fatty acid synthase (FAS), sterol regulatory element binding protein‐1c (SREBP‐1c) in the IPF and hepatopancreas reached the lowest in 2.0% group. Overall, dietary ALA could promote n‐3 highly unsaturated fatty acids (HUFAs) synthesis and suppress the accumulation of lipid by decreasing the expression of related genes and promoting the apoptosis in IPF.     

The future of genetic engineering to provide essential dietary nutrients and improve growth performance in aquaculture: Advantages and challenges

Advancements in gene technology in recent years have been driving the aquaculture industry forward. Improvements in growth performance, feed efficiency, and omega‐3 content are goals of the industry that could capitalize on applications of genetic engineering. One of the major challenges in the industry is to reduce the use of fish meal and oil, to improve the environmental and economic sustainability of aquaculture. The recent development of genetically engineered feed ingredients is one potential solution to the looming problem of fish meal and oil dependency. Furthermore, the development of transgenic fish has potential to improve production efficiency and other future desirable characteristics that relate to feed utilization and product quality. New gene technologies are beginning to revolutionize how we produce our food, and in aquaculture, will ultimately reduce pressure on wild fish stocks, help to preserve natural aquatic ecosystems, and improve nutritional profiles of farmed fish for human consumption. The purpose of this review is to provide an update on the current applications of genetic engineering technology to improve aquaculture through nutrition, including the development and use of transgenic feed ingredients, transgenic fish, and ultimately their impacts on nutrition, product quality, and consumers.     

Knockdown of HMGA2 gene inhibits TGF-β1-induced human embryonic lung fibroblast growth and collagen synthesis

AIM: To investigate the effects of high mobility group A2(HMGA2) gene knockdown on the cell viability, apoptosis, collagen synthesis and oxidative stress of human embryonic lung fibroblast (HELF) induced by transforming growth factor-β1 (TGF-β1). METHODS: The HELF were divided into blank group, TGF-β1 group,negative control (NC) group and HMGA2 siRNA(si-HMGA2) group. The protein levels of HMGA2, AKT and p-AKT were determined by Western blot. The cell viability and apoptotic rate was analyzed by MTT assay and flow cytometry,respectively. The mRNA expression of collagen I (COL-Ⅰ) and COL-Ⅲ was detected by RT-qPCR. DCFH-DA was used to detect the content of reactive oxygen species (ROS). RESULTS: Compared with blank group, the protein levels of HMGA2 and p-AKT, the cell viability, the mRNA expression of COL-Ⅰ and COL-Ⅲ in TGF-β1 group were significantly increased, but the apoptotic rate and ROS level were significantly decreased (P<0.05). Compared with TGF-β1 group, the protein levels of HMGA2 and p-AKT, the cell viability, the mRNA expression of COL-Ⅰ and COL-Ⅲ in si-HMGA2 group were significantly decreased, but the apoptotic rate and ROS level were significantly increased (P<0.05). CONCLUSION: Knockdown of HMGA2 gene expression decreases the viability and collagen synthesis, and promotes apoptosis and ROS production of human embryonic lung fibroblasts induced by TGF-β1. The mechanism may be related to down-regulation of PI3K/AKT signaling pathway.     

miR-140-3p inhibits viability,invasion and migration of non-small-cell lung cancer A549 cells by targeting PD-L1

AIM: To explore the target relationship between microRNA-140-3p (miR-140-3p) and programmed cell death ligand 1 (PD-L1) and their effect on the viability, migration and invasion of non-small-cell lung cancer A549 cells.METHODS: RT-qPCR was used to detect the miR-140-3p expression in HLF-1, A549 and H1299 cells, and then the A549 cells with the most significant difference were selected as the subsequent research object. TargetScan software and dual-luciferase reporter assay were performed to predict and confirm the target relationship between miR-140-3p and PD-L1. RT-qPCR and Western blot were used to determine the effects of miR-140-3p mimic and inhibitor on PD-L1 expression level. MTT assay was used to detect the viability of A549 cells. Transwell assay was performed to detect the migration and invasion abilities of the A549 cells.RESULTS: miR-140-3p was significantly down-regulated in the A549 cells and H1299 cells (P<0.05). Transfection with miR-140-3p mimic decreased the expression of PD-L1 and inhibited the viability, migration and invasion of the A549 cells. Transfection with pcDNA3.0-PD-L1 reversed the inhibitory effect of miR-140-3p on the viability, migration and invasion of the A549 cells.CONCLUSION: miR-140-3p inhibits the viability, migration and invasion of A549 cells by targeting PD-L1.     

Cepharanthine induces autophagy via PI3K/AKT/mTOR signaling pathway in ovarian cancer SKOV3 cells

AIM: To investigate the autophagy of human ovarian cancer SKOV3 cells induced by cepharanthine and to explore its mechanism. METHODS: The effect of cepharanthine on the viability of ovarian cancer SKOV3 cells was measured by CCK-8 assay. The SKOV3 cells were treated with cepharanthine, and then the formation of autophagosome was observed with acridine orange staining under fluorescence microscope. The protein levels of LC3, AKT, p-AKT, mTOR, p-mTOR and GAPDH in the SKOV3 cells treated with cepharanthine were determined by Western blot.RESULTS: Cepharanthine significantly inhibited the viability of ovarian cancer SKOV3 cells in a dose-dependent manner (P<0.05). The number of the intracellular acidic autophagosomes with bright red fluorescence was significantly increased after cepharanthine treatment in the SKOV3 cells. The expression of LC3-Ⅱ in SKOV3 cells was significantly enhanced after cepharanthine treatment. Furthermore, treatment with cepharanthine in the SKOV3 cells also resulted in a significant down-regulation of phosphorylated form of AKT and mTOR (P<0.01), while the total protein level was not changed. Combination of cepharanthine and 3-methyladenine resulted in a substantial decrease in the cell viability compared with using cepharanthine alone.CONCLUSION: Cepharanthine significantly inhibits the growth of human ovarian cancer SKOV3 cells and induces the autophagy, which may be correlated with down-regulation of PI3K/AKT/mTOR signaling pathway.