Identification of anthocyanin biosynthesis related microRNAs and total microRNAs in Lonicera edulis by high-throughput sequencing

miRNAs are important regulators of plant gene expression. There are few studies on the regulation of miRNAs in Lonicera edulis. We used high-throughput sequencing technology to analyse miRNAs in L. edulis, aiming to identify miRNAs and elucidate their function in L. edulis. In the present study, we employed the high-throughput sequencing technology to profile miRNAs in L. edulis. A total of 51,819,072 small RNA tags with sizes ranging from 18 to 30 nt were obtained, indicating that L. edulis have a large and diverse small RNA population. Bioinformatic analysis identified 507 mature miRNAs, and 16 predicted novel miRNAs that are likely to be unique to L. edulis. Three miRNAs related to anthocyanin biosynthesis were locked by gene ontology (GO) analysis and target gene analysis. The selected three miRNAs are relatively high in the expression of L. edulis. Some of the previous studies have studied these types of miRNAs involved in the anthocyanin metabolism pathway in fruits. Among them, expression profiles of three conserved miRNAs were validated by stem loop qRT-PCR. Further, the potential target genes of conserved and novel miRNAs were predicted and subjected to GO annotation. Enrichment analysis of the GO-represented biological processes and molecular functions revealed that these target genes were potentially involved in a wide range of metabolic pathways and developmental processes. In particular, different families of miRNAs can directly or indirectly regulate anthocyanin biosynthesis. In recent years, the research on miRNAs has become more and more clear, but the research on miRNAs involved in the regulation of anthocyanin synthesis of L. edulis is still lagging. This study provides a useful resource for further elucidation of the functional roles of miRNAs during fruit development and ripening

LINC24065 is a monoallelically expressed long intergenic noncoding RNA located in the cattle DLK1–DIO3 cluster

Long noncoding RNAs (lncRNAs) are important regulators of biological processes, and regulate genomic imprinting in cis and/or trans to induce monoallelic expression with parent-origin-specific pattern. DLK1–DIO3 domain is one of the largest imprinted clusters in mammals, and maternally expressed noncoding RNAs of this region is related to the pluripotency of the embryonic stem cells. Previously, we sequenced the cDNA of two maternally expressed noncoding RNAs, MEG8 and MEG9, and mapped a lncRNA (LINC24061) between the two genes in the cattle DLK1–DIO3 domain on chromosome 21. In this study, we identified LINC24065, a novel long intergenic noncoding RNA (lincRNA), which was also located between MEG8 and MEG9. We identified four variants of LINC24065 (LINC24065-v1, LINC24065-v2, LINC24065-v3 and LINC24065-v4) that were a result of alternative splicing from 18 exons. LINC24065-v1 and LINC24065-v2 showed tissue-specific expression patterns in adultbovine tissues, and LINC24065-v3 and LINC24065-v4 were detected in all eight analysed tissues (heart, liver, spleen, lung, kidney, skeletal muscle, adipose and brain). Using single-nucleotide polymorphism (SNP)-based method, LINC24065 was identified to have monoallelic expression in adult tissues, suggesting that it is imprinted in cows. These results provide a foundation for further investigation about whether LINC24065 plays a role in regulating imprinting of the DLK1–DIO3 domain.

The application of three-dimensional reconstruction-based Micro-CT in the research of root canal treatment / 口腔疾病防治

@#Important guarantees of the success of root canal therapy include being familiar with the anatomical features of root canal system, good root canal preparation and complete root canal filling. With the development of medical imaging technology, three-dimensional reconstruction technology has been widely applied in root canal morphology, root canal preparation, root canal filling quality, root canal retreatment efficiency, three-dimensional finite elements and other related research. The reconstructed 3D images can be observed qualitatively and quantitatively from different angles and levels, which aid in the understanding of root canal anatomy and evaluation of the operation effect on all aspects of root canal therapy, providing important guiding significance for clinical operation. The application of 3D reconstruction technology based on micro-CT in the research of root canal anatomy, root canal preparation, root canal filling, root canal retreatment efficiency and three-dimensional finite element analysis are reviewed. The results of a literature review showed that the resolution of micro-CT is high. Furthermore, three-dimensional reconstruction can accurately display the fine anatomical morphology of a root canal, accurately measure the volume and morphological changes of root canal before and after root canal preparation, accurately measure the incidence and volume of the void after root canal filling and accurately calculate the volume changes of the root canal filling materials before and after root canal retreatment. Micro-CT is the gold standard for the morphological study of the root canal system and quality analysis of root canal preparation and filling. After three-dimensional reconstruction with micro-CT scans, a three-dimensional finite element model can be established to simulate the process of root canal therapy and stress analysis. This method can be used to analyze the stress distribution of root canal preparation instruments and the influence of root canal preparation on the stress distribution of tooth tissue and the root canal wall in the process of root canal filling. Then, suitable operation methods can be selected to prevent instrument breakage and root fracture. The application of three-dimensional reconstruction-based micro-CT is of great value in the evaluation of root canal morphology and treatment.

Effects of different root canal filling stop on quality of root canal filling in root canals obturated with GuttaFlow / 口腔疾病防治

Objective @# To investigate the effects of different root canal filling stop on quality of root canal filling and apical sealing in root canals obturated with GuttaFlow. @* Methods@#60 teeth were randomly divided into three groups, using different root canal filling stops to shape the root canals with MTwo (25/06) file. All root canals were obturated with Gutta Flow, and the overfilling of the root canals were recorded and evaluated by X-ray. And the apical microleakage of teeth was evaluated by transparent teeth technique.@*Results@#The roots were prepared with MTwo (25/06) as master apical file, the overfilling rate of the root canals in root canal filling stop was higher as the distance from the apex was shorter, but there was no significant difference. The under-filling rate of the root canals in root canal filling stop was higher as the distance from the apex was longer. And the under-filled root canals in root canal filling stop 0.5 mm from the apex showed a statistically significant difference with 2 mm. The mean dyeing penetration length in 0.5 mm and 1 mm group was significantly shorter than 2 mm group. @*Conclusion @#A suitable root canal filling stop could improve the quality of root canal filling in root canals obturated with GuttaFlow.

Effects of Chitosan on the Secretion of Cytokines and Expression of Inducible Nitric Oxide Synthase mRNA in Peritoneal Macrophages of Broiler Chicken

An in vitro experiment was conducted to study the effects of chitosan on the secretion of cytokines and expression of inducible nitric oxide synthase mRNA in peritoneal macrophages of broiler chicken. In the experiment, peritoneal macrophages were incubated for 24 h in culture medium supplemented with 0 (control), 40, 80, 160 and 320 µg/mL chitosan. The results showed that chitosan tended to increase quadratically the levels of interleukin-1 (P = 0.093) and interleukin-2 (P = 0.106) in the culture fluid of peritoneal macrophage. Chitosan also significantly enhanced inducible nitric oxide synthase mRNA expression of peritoneal macrophage in a quadratic dose-dependent manner (P < 0.05) and tended to promote quadratically the secretion of nitric oxide (P = 0.053) and inducible nitric oxide synthase (P = 0.157) in peritoneal macrophages. This result implied that one of the mechanisms by which chitosan modulated immune functions in chickens might be chitosan activating expression of inducible nitric oxide synthase and then improving the secretion of nitric oxide.