High-throughput miRNA sequencing and identification of a novel ICE1-targeting miRNA in response to low temperature stress in Eucalyptus camaldulensis.

MicroRNAs (miRNAs) play a crucial role in the growth, development, morphogenesis, signal transduction, and stress response in plants. The ICE (Inducer of CBF expression)-CBF (C-repeat binding factor)-COR (Cold-regulated gene) regulatory cascade is an important signalling pathway in plant response to low temperature stress, and it remains unknown whether this pathway is regulated by miRNAs. In this study, high-throughput sequencing was employed for predicting and identifying the miRNAs that were likely to target the ICE-CBF-COR pathway in Eucalyptus camaldulensis. A novel ICE1-targeting miRNA, eca-novel-miR-259-5p (nov-miR259), was further analysed. A total of 392 conserved miRNAs and 97 novel miRNAs were predicted, including 80 differentially expressed miRNAs. Of these, 30 miRNAs were predicted to be associated with the ICE-CBF-COR pathway. The full-length of mature nov-miR259 was 22 bp and its precursor gene was 60 bp in length, with a typical hairpin structure. The RNA ligase-mediated 5' amplification of cDNA ends (5'-RLM-RACE) and Agrobacterium-mediated tobacco transient expression assays demonstrated that nov-miR259 could cleave EcaICE1 in vivo. Moreover, qRT-PCR and Pearson's correlation analysis further revealed that the expression levels of nov-miR259 were almost significantly negatively correlated with those of its target gene, EcaICE1, and the other genes in the ICE-CBF-COR pathway. We first identified the nov-miR259 as a novel ICE1-targeting miRNA, and the nov-miR259-ICE1 module may be involved in regulating the cold stress response in E. camaldulensis.

The Impact of a Patient Education Package on Outcomes of Pain Management Following Orthopaedic Surgery in a Tertiary Hospital in Malaysia

Patient’s belief towards pain management may affect pain management outcomes and quality of life. The main aim of the present study was to determine the impact of a pre-operative pain education package towards pain belief among patients undergoing orthopaedic surgery in a tertiary hospital. A one-group pre-test post-test design study was conducted on orthopaedic surgery patients. Thirty respondents were recruited and pre-operative pain education was administered individually before surgery. Pre-operative and post-operative pain belief, management scores and side effects were measured using the Barrier Questionnaire (BQ-13). The results reported significant differences between pre-test scores (Mean = 41.87, Standard Deviation = 11.467) and post-test scores (Mean=34.80, Standard Deviation=13.026) of pain belief (t = 2.84, p = 0.004). There were also significant differences between pre-test scores (Mean = 37.10, Standard Deviation = 10.610) and post-test scores (Mean=30.80, Standard Deviation = 11.424) of pain management (t = 3.856, p = 0.0005). Respondent’s gender (t = -2.403, p = 0.023) and ethnicity (F = 5.038, p=0.014) reported significant differences with p value 0.05). There was positive impact of the pain education package towards pain belief and painmanagement among respondents who underwent orthopaedics surgery in a tertiary hospital. Reinforcement of pain educational program is pivotal in order to achieve optimal post-operative pain management.