ABSTRACT
@#AIM: To detect the relative expression levels of microRNA-27a(miR-27a)and nuclear factor erythroid-2-related factor 2(NRF2)in serum of patients with age-related macular degeneration(ARMD)bleeding, and to explore the correlation between the expression levels and the prognosis of ARMD bleeding. <p>METHODS: A retrospective case series observation was carried out.From June 2018 to October 2019, 80 patients with ARMD bleeding who were treated in our hospital were selected as ARMD bleeding group, and 80 healthy people who had routine examination in our hospital were selected as control group. The relative expression levels of miR-27a and NRF2 were detected by real-time fluorescent quantitative PCR(qRT-PCR), the diagnostic value of serum miR-27a and NRF2 expression for ARMD bleeding was evaluated by receiver operating characteristic curve(ROC). The incidence of poor prognosis was analyzed; in addition, Logistic regression was used to analyze the influencing factors of poor prognosis in patients with ARMD bleeding. <p>RESULTS: The relative expression level of miR-27a in serum of ARMD bleeding group was significantly higher than that of control group(<i>P</i><0.01), and the relative expression level of NRF2 mRNA in serum was significantly lower than that in control group(<i>P</i><0.01). ROC results showed that the AUC of serum miR-27a and NRF2 in the diagnosis of ARMD bleeding was 0.867 and 0.820 respectively, and the cutoff value was 1.10 and 1.08 respectively, at this time, the corresponding sensitivity was 71.3% and 91.3%, and the specificity was 90.0% and 63.7%, respectively. The AUC of serum miR-27a combined with NRF2 in the diagnosis of ARMD bleeding was 0.912, and the corresponding sensitivity and specificity were 86.3% and 85.0%, respectively. The incidence of poor prognosis in high miR-27a group was significantly higher than that in low miR-27a group(<i>P</i><0.05); and the incidence of poor prognosis in high NRF2 group was significantly lower than that in low NRF2 group(<i>P</i><0.05). Logistic analysis showed that the high expression of serum miR-27a was an independent risk factor for poor prognosis in patients with ARMD bleeding, and the high relative expression of NRF2 in serum was a protective factor for the poor prognosis of patients with ARMD bleeding. <p>CONCLUSION: The relative expression level of miR-27a in the serum of patients with ARMD hemorrhage is significantly increased, and the relative expression level of NRF2 is significantly decreased, both of them have certain diagnostic value for ARMD bleeding, and their relative expressions are closely related to the prognosis of patients, which is suggested that miR-27a and NRF2 can be used as potential biological indexes for early diagnosis and prognosis evaluation of ARMD bleeding.
ABSTRACT
Objective To obtain a full-length cDNA of squalene synthase gene from Hedera helix (HhSS) by cloning technique, and to carry out bioinformatics analysis and expression analysis. Methods Primers were designed based on H. helix transcriptome data, and HhSS was cloned by using RACE technologies. DNAMAN, PROTPARAM, TMHMM, PSORT, ScanProsite, SOPMA, and SWISS-MODEL were used for analysis of sequence and physical and chemical properties and domain of encoded protein. The relative expression of HhSS was detected by qRT-PCR. Results The cDNA sequence of HhSS (GenBank accession number: KX056078) was 1 889 bp which obtained by RACE cloning. It contained an ORF from 248 bp to 1 477 bp and a 5'UTR with 247 bp and a 3'UTR with 412 bp. It encoded a 409-amino-acid protein with a molecular weight of 46 800and an isoelectric point (pI) of 5.68. The HhSS protein had the characteristic domain and transmembrane region of plant SS protein and closer relationship with Eleutherococcus senticosus, Panax ginseng et al. The qRT-PCR results indicated that it had positive correlation between relative expressing level of HhSS gene and contents of saponins in H. helix leaves. Conclusion The cloning and expression analysis results of HhSS provide a theoretical and technical basis for elucidating the role of HhSS in saponins biosynthetic pathway and metabolic regulation.