In biological research, chicken embryos are a classic experimental model for the exploration of the embryonic development and cell differentiation.  Transferring exogenous substances into chicken embryos for producing medical antibodies has been widely used in the production practice.  However, there are few studies about the effect of the different injection site and dosage on chicken embryos.  The aim of this study was to explore the effects of different injection sites and dosages on chicken embryo hatching rate and development, so as to provide a basis for further studies using the chicken embryo model.  Freshly laid eggs (Rugao yellow chicken) were injected with different doses of saline at the tip, equatorial plane and the blunt end of the egg shell, respectively.  Egg hatching rate was recorded and compared among injection sites and different doses.  A trypan blue stain was also injected at the aforementioned sites and the growth of chicken embryos was observed.  The SPSS (statistical package for the social science) software was used to analyze the relationship between the chicken eggs hatching rate and the different injection sites or the different dosages.  The experimental results showed that there were significant differences on egg hatching rates among the different injection sites and doses (P<0.05).  The hatchability of the blunt end injection group was significantly higher than that of the other two sites.  The egg hatching rate decreased with increased saline doses.  The egg hatching rate of the 100 µL saline injection group was higher than the 200 and 300 µL dosage groups.  Ultimately, we suggest that the optimal chicken embryo injection process is during early development, at the blunt end site with a dose less than 100 µL to minimize damage to the egg.

    High-throughput sequencing has identified a large number of sense-antisense transcriptional pairs, which indicates that these genes were transcribed from both directions. Recent reports have demonstrated that many antisense RNAs, especially lncRNA (long non-coding RNA), can interact with the sense RNA by forming an RNA duplex. Many methods, such as RNA-sequencing, Northern blotting, RNase protection assays and strand-specific PCR, can be used to detect the antisense transcript and gene transcriptional orientation. However, the applications of these methods have been constrained, to some extent, because of the high cost, difficult operation or inaccuracy, especially regarding the analysis of substantial amounts of data. Thus, we developed an easy method to detect and validate these complicated RNAs. We primarily took advantage of the strand specificity of RT-PCR and the single-strand specificity of S1 endonuclease to analyze sense and antisense transcripts. Four known genes, including mouse β-actin and Tsix (Xist antisense RNA), chicken LXN (latexin) and GFM1 (G elongation factor, mitochondrial 1), were used to establish the method. These four genes were well studied and transcribed from positive strand, negative strand or both strands of DNA, respectively, which represented all possible cases. The results indicated that the method can easily distinguish sense, antisense and sense-antisense transcriptional pairs. In addition, it can be used to verify the results of high-throughput sequencing, as well as to analyze the regulatory mechanisms between RNAs. This method can improve the accuracy of detection and can be mainly used in analyzing single gene and was low cost.

Building ontology is a fundamental but also hard work. Collaborative ontology editing tools can make ontology development more efficiently. In this paper, the important features of collaborative ontology development were analyzed, and several tools such as AGROVOC Concept Server Workbench (ACSW), Collaborative Protégé and WebProtégé were studied. Besides, some comparisons among them from several aspects were made and some prospects for the further improvement of these tools were given. Finally, we show it is a good way to build agricultural ontology with these tools collaboratively and simultaneously.