The bioflocs technology (BFT) for shrimp production has been proposed as a sustainable practice capable of reducing environmental impacts and preventing pathogen introduction. The microbial community ...associated with BFT not only detoxifies nutrients, but also can improve feed utilization and animal growth. Biofloc system contains abundant number of bacteria of which cell wall consists of various components such as bacterial lipopolysaccharide, peptidoglycan and β‐1, 3‐glucans, and is known as stimulating nonspecific immune activity of shrimp. Bioflocs, therefore, are assumed to enhance shrimp immunity because they consume the bioflocs as additional food source. Although there are benefits for having an in situ microbial community in BFT systems, better understanding on these microorganisms, in particular molecular level, is needed. A fourteen‐day culture trial was conducted with postlarvae of Litopenaeus vannamei in the presence and absence of bioflocs. To determine mRNA expression levels of shrimp, we selected six genes (prophenoloxidase1, prophenoloxidase2, prophenoloxidase activation enzyme, serine proteinase1, masquerade‐like proteinase, and ras‐related nuclear protein) which are involved in a series of responses known as the prophenoloxidase (proPO) cascade, one of the major innate immune responses in crustaceans. Significant differences in shrimp survival and final body weights were found between the clear water and in the biofloc treatments. mRNA expression levels were significantly higher in the biofloc treatment than the clear water control. These results suggest that the presence of bioflocs in the culture medium gives positive effect on growth and immune‐related genes expression in L.vannamei postlarvae.
Recombination activating genes (RAGs) mediates the process of rearrangement and somatic recombination (V(D)J) to generate different antibody repertoire. Studies on the expression pattern of adaptive ...immune genes during ontogenic development are crucial for the formulation of fish immunization strategy. In the present study, Nile tilapia was taken to explore the relative expression profile of RAG genes during their developmental stages. The developmental stages of Nile tilapia, i.e., unfertilized egg, 0, 2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28 and 30 days post-hatch (dph) and kidney, blood, gill, liver and spleen tissues from adult fish were collected and the cDNA synthesis was carried out. Gene specific primers for RAG-1 and RAG-2 of Nile tilapia were designed and their annealing temperature (Tm) was optimized by gradient PCR. Consequently, PCR was performed to confirm the specific amplification of RAG-1 and RAG-2 genes. Quantitative real-time PCR (qRT-PCR) gene expression of RAG-1 and RAG-2 were noticed in all the developmental stages; however, a significant increase was observed after 12 dph and peaked at 24 dph, followed by a gradual decrease until 30 dph. Tissue-specific gene expression profiling revealed that the highest expression of RAG-1 and RAG-2 was observed in the kidney, followed by spleen, gill, liver and blood. The findings of the study explored the suitable timing of lymphoid maturation that could be technically used for the adoption of strategies to improve disease resistance of fish larvae for mitigating larval mortality.
•The expression of RAG-1 and RAG-2 starts from the twelve days post hatch.•Kidney shows the abundance of RAG-1 mRNA.•The expression of RAG-1 is higher than RAG-2 across the tissues.
There is increasing evidence that circular RNA (circRNA) are involved in cancer development, but the regulation and function of human circRNA remain largely unknown. In this study, we demonstrated ...that ZKSCAN1, a zinc finger family gene, is expressed in both linear and circular (circZKSCAN1) forms of RNA in human hepatocellular carcinoma (HCC) tissues and cell lines. Here, we analyzed a cohort of 102 patients and found that expression of both ZKSCAN1mRNA and circZKSCAN1 was significantly lower (P < 0.05) in the HCC samples compared with that in matched adjacent nontumorous tissues by reverse transcription PCR (RT‐PCR). The low expression level of ZKSCAN1 was only associated with tumor size (P = 0.032), while the cirZKSCAN1 levels varied in patients with different tumor numbers (P < 0.01), cirrhosis (P = 0.031), vascular invasion (P = 0.002), or microscopic vascular invasion (P = 0.002), as well as with the tumor grade (P < 0.001). Silencing both ZKSCAN1mRNA and circZKSCAN1 promoted cell proliferation, migration, and invasion. In contrast, overexpression of both forms of RNA repressed HCC progression in vivo and in vitro. Silencing or overexpression of both forms of RNA did not interfere with each other. RNA‐seq revealed a very different molecular basis for the observed effects; ZKSCAN1mRNA mainly regulated cellular metabolism, while circZKSCAN1 mediated several cancer‐related signaling pathways, suggesting a nonredundant role for ZKSCAN1mRNA and circRNA. In conclusion, our results revealed two post‐translational products (ZKSCAN1mRNA and circZKSCAN1) that cooperated closely with one another to inhibit growth, migration, and invasion of HCC. cirZKSCAN1 might be a useful marker for the diagnosis of HCC.
Circular RNA have recently been shown to play a regulatory role in disease. We investigated the role of circRNA transcribed from ZKSCAN1, a member of the zinc finger family. Our results revealed two post‐translational products (ZKSCAN1 mRNA and circZKSCAN1) that cooperated closely with one another to inhibit cancer growth migration and invasion, but which affected different signaling pathways.
Colorectal cancer (CRC) has been one of the most commonly diagnosed cancers in global. The differential expression profiles of microRNAs (miRNAs) in CRC plasma of patients have the potential to serve ...as a diagnostic biomarker. We conducted a four-stage study to identify the potential plasma miRNAs for CRC detection. In the initial screening phase, Exiqon panel (miRCURY-Ready-to-Use-PCR-Human-panel-I + II-V1.M) including 3 CRC pools and 1 normal controls (NCs) pool were applied to acquire miRNA profiles. In the training stage (30 CRC VS. 30 NCs) and testing stage (79 CRC VS. 76 NCs), quantitative real-time polymerase chain reaction (qRT-PCR) was utilized to conduct candidate miRNA profiles. Then the identified miRNAs were verified in external validation stage (30 CRC VS. 26 NCs). Expression levels of identified miRNAs were assessed in tissue samples (24 pairs) and plasma exosomes (18 CRC VS. 18 NCs). Receiver operating characteristic (ROC) curves were constructed to evaluate the diagnostic accuracy. Seven miRNAs (miR-103a-3p, miR-127-3p, miR-151a-5p, miR-17-5p, miR-181a-5p, miR-18a-5p and miR-18b-5p) were significantly overexpressed in CRC compared with NCs. Area under the ROC curve of the seven-miRNA signature was 0.762, 0.824 and 0.895 for the training, testing and the external validation stages, respectively. Additionally, miR-103a-3p, miR-127-3p, miR-17-5p and miR-18a-5p were discovered significantly up-regulated in CRC tissues; while miR-17-5p, miR-181a-5p, miR-18a-5p and miR-18b-5p were significantly elevated in CRC plasma exosomes. In conclusion, we established a seven-miRNA signature in the peripheral plasma for CRC detection.
•We investigated plasma miRNA profiles of CRC in a four-stage process and present a seven-miRNA signature in discriminating CRC.•We found that miR-103a-3p, miR-127-3p, miR-17-5p and miR-18a-5p were significantly up-regulated in CRC tissues.•We found that miR-17-5p, miR-181a-5p, miR-18a-5p and miR-18b-5p were significantly elevated in CRC plasma exosomes.•We discovered that plasma miR-26a-5p was only up-regulated in colon cancer patients but not in rectal cancer.•We found none of the seven miRNAs demonstrated significant different expression in patients with stage III + IV compared to those with stage I + II.•We found that no difference of the above seven plasma miRNA between left-sided and right-sided CRC.
Exosomal microRNAs (miRNAs) have great potentials as a novel biomarker to predict lung cancer. We applied a miRNA microarray to identify aberrantly expressed serum exosomal miRNAs as candidate ...biomarkers for patients with lung adenocarcinoma (LUAD). Compared with the normal control, 31 exosomal miRNAs were found to be upregulated and 29 exosomal miRNAs were downregulated in the serum of LUAD respectively. Then, 10 dysregulated exosomal miRNAs expression levels in serum were further validated via qRT‐polymerase chain reaction. Notably, exosomal miR‐7977 was highest expressed and miR‐98‐3p was lowest expressed in the patients with LUAD, and exosomal miR‐7977 showed significant correlation with the N stage and TNM stage with patients with LUAD (P < .05). Receiver operating characteristic curve showed that the abundant level of exosomal miR‐7977 may predict LUAD with an area of under the curve (AUC) of 0.787. In comparison with exosomal miR‐7977, exosomal miR‐98‐3p had a smaller area (0.719). The combination of exosomal miR‐7977 and miR‐98‐3p improved the AUC to 0.816. Furthermore, in vitro experiments revealed that inhibition of miR‐7977 enhanced the proliferation, invasion, and inhibited apoptosis in A549 cells, the opposite results were performed by miR‐7977 mimics. In conclusion, exosomal miR‐7977 was identified as a novel biomarker for patients with LUAD and may play as a tumor suppressor in lung cancer.
We were the first time to provides evidence that serum exosomes‐derived miR‐7977 is a novel promising diagnostic biomarker for lung adenocarcinoma and may play a tumor suppressor in lung cancer.
This study aimed to identify differential circular RNA (circRNA) in the plasma exosomes of patients with lung adenocarcinoma (LUAD) using high‐throughput sequencing. First, exosomes were isolated ...using an exosome isolation kit and confirmed by Western blotting, transmission electron microscopy, and NanoSight Assay. Subsequently, plasma circRNA expression profiles were screened by high‐throughput sequencing and confirmed by fluorescence quantitative real‐time polymerase chain reaction (qRT‐PCR) and Sanger sequencing. Finally, the circRNA‐miRNA‐mRNA network was performed to forecast the potential function of circRNAs. The result of high‐throughput sequencing data documented that 182 differentially expressed exosomal circRNAs in all were screened, which included 105 that were upregulated and 78 that were downregulated in LUAD patients plasma compared with controls. The four upregulated circRNAs including circ_0001492, circ_0001346, circ_0000690, and circ_0001439 were identical to the sequencing data by qRT‐PCR, and their latent circRNA‐miRNA‐mRNA interactions were exhibited. Taken together, our study firstly revealed the altered exosomal circRNA expression from plasma samples in patients with LUAD and supports the need for exploring their potential as biomarkers and the pathological effects of lung cancer.
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The comprehensive expression profiles of circular RNAs (circRNAs) in plasma exosomes of early‐stage lung adenocarcinoma (LUAD) are firstly illustrated.
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Four exosomal circRNAs are definitely dysregulated based on quantitative real‐time polymerase chain reaction.
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Potential microRNA and targets gene are predicted for these validated circRNAs.
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CircRNAs of exosomes may act as novel noninvasive biomarkers for LUAD.
Proteins participate in defense mechanisms after a pathogen attacks a host. Pathogenesis-related (PR) proteins in wheat plants play the important role of defense against Puccinia striiformis f. sp. ...Tritici. This study uses various analytic methods to describe the expression of five (PR) protein genes (PR1, PR2, PR4, PR9 and PR10) and genetic polymorphism among sixteen wheat genotypes to understand the mechanisms underlying of PR genes in Egyptian wheat compared with the Yr genotype. The PR protein genes were semi-quantitatively and quantitatively assessed in four selected wheat genotypes; two resistant (Misr-3 and Yr-29) and two susceptible (Sids-12 and Avocet-S). Reverse transcriptase PCR analysis results indicated that all PR protein genes exhibited different expression levels among the four studied genotypes and overexpression at the resistant genotypes in the different time points of Pst inoculation. Quantitative real-time polymerase chain reaction (qRT-PCR) analysis indicated that the expression of PR protein genes was elevated in adult plant resistant (APR) genotypes. Plant stage in combination with the infection process by stripe rust provided even greater upregulation in the expression of PR genes and enhanced induction of defense enzyme activities in the same sample at different time points for various wheat genotypes.
•Sixteen wheat genotypes were evaluated under artificial infection with stripe rust, Misr-3 was the most resistant genotype.•Five PR protein genes were used to evaluate and produce unique DNA profiles for the studied genotypes.•Expression of PR protein genes was more effective in adult plant than the seedling.•Induction of PR protein genes was correlated with CAT and POX defense enzyme activities at the same time point.
Circulating microRNAs (miRNAs) have the potential to become reliable and noninvasive biomarkers for ovarian cancer (OC) diagnosis; however, the conventional miRNAs detection techniques exhibit ...enduring limitations of low sensitivity and specificity. Graphene oxide (GO), a novel nanomaterial, is at the forefront of material design for extensive biomedical applications. Owing to the excellent water affinity and single-stranded DNA (ssDNA) adsorption characteristics of GO, we designed and developed a GO-based qRT-PCR assay for the detection of miRNAs associated with OC. In the GO-based qRT-PCR system, GO could significantly improve the sensitivity and specificity of the qRT-PCR assay by noncovalently interacting with primers and ssDNA and reducing the occurrence of non-specific amplification. Moreover, the detection of miRNAs associated with OC confirmed that GO-based qRT-PCR assay could differentiate benign ovarian tumors from OC (sensitivity, 0.91; specificity, 1.00). Collectively, these findings provide robust evidence that GO-based qRT-PCR assay can be effectively used as a promising method to detect miRNAs for the screening of OC patients.
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•The graphene oxide-based qRT-PCR assay was used to determine miRNAs in plasma.•The graphene oxide-based qRT-PCR assay lowered the detection limit and eliminated non-specific amplification.•High sensitivity and specificity diagnostic method of ovarian cancer was established by the graphene oxide-based qRT-PCR.
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