Hsa_circ_0001658 accelerates the progression of colorectal cancer through miR-590-5p/METTL3 regulatory axis.

BACKGROUND: As reported, multiple circular RNAs (circRNAs) interfere with colorectal cancer (CRC) progression. Here, circRNA_0001658 (circ_0001658) is focused on studying how it works in CRC. AIM: Clarify the expression pattern, biological function, and underlying mechanism of circ_0001658 of CRC tumorigenesis. METHODS: In CRC-related chip data retrieved using the database named Gene Expression Omnibus, different expressions of circRNAs between CRC and normal tissue samples were identified. Quantitative Real-time PCR and Western blot ensured the analysis on circ_0001658, microRNA-590-5P (miR-590-5p), and methyltransferase-like 3 (METTL3) mRNA expressions in tissues and cells. Cell counting kit-8 and flow cytometry were used to detect cell proliferation, apoptosis and migration. The targeting relations between circ_0001658, miR-590-5p, and METTL3 mRNA 3'-untranslated region were under the verification of bioinformatics prediction and dual luciferase-based reporter gene assays. Gene Ontology and Kyoto Encyclopedia of Genes and Genomes enrichment analysis were employed on the downstream targets of miR-590-5p using the Database for Annotation, Visualization and Integrated Discovery database. RESULTS: Circ_0001658 and METTL3 mRNA was elevated in CRC tissues and cells, whereas miR-590-5p was decreased. Circ_0001658 overexpression promoted the proliferation of HT29 cells, inhibited apoptosis, and accelerated the cell cycle. In SW480 cells, knocking down circ_0001658 had the opposite effect. Circ_0001658 could specifically bind to miR-590-5p and negatively modulate its expressions; METTL3 is a miR-590-5p target that can be positively regulated by circ 0001658. Circ 0001658 was inversely associated with miR-590-5p expression while positively with METTL3 expressions. CONCLUSION: Circ_0001658 regulates the miR-590-5p/METTL 3-axis to increase CRC cell growth and decrease apoptosis.

Prediction of frequency-specific hearing threshold using chirp auditory brainstem response in infants with hearing losses.

OBJECTIVES: To investigate the clinical usefulness of the LS-chirp auditory brainstem response for estimation of behavioral thresholds in young children with mild to severe hearing losses. METHODS: 68 infants (136 ears) aged 6-12 months (mean age=9.2 months) with bilateral mild to severe hearing losses were studied at Children's Hospital of Fudan University. In all cases, the children were referred for LS-chirp ABR and visual reinforcement audiometric (VRA) measurements. The low-frequency band chirp (LF-chirp) thresholds (frequency band=0.1-0.85kHz) were compared to the average VRA thresholds (frequency band=0.25-0.5kHz), whereas the high-frequency band chirp (HF-chirp) thresholds (frequency band=1-10kHz) were compared to the average VRA thresholds (frequency band=1-4kHz) using statistical correlation coefficient values. RESULTS: The LS-chirp ABR thresholds are very close to behavioral hearing levels. The mean differences between chirp-ABR and VRA thresholds were within 5dBHL for all measurements. The smallest mean threshold difference (<3dBHL) was obtained for the severe hearing loss group. The correlation coefficient values (r) were 0.97 at low-frequency and high-frequency bands. For each carrier frequency, the best correlations between chirp-ABR thresholds and VRA thresholds were obtained at VRA frequency of 0.25kHz/LF-chirp (r=0.98) and VRA frequency of 1kHz/HF-chirp (r=0.98). CONCLUSIONS: This study demonstrates the effectiveness using chirp-ABR predicted frequency-specific thresholds, especially of low and middle frequencies. LS-chirp ABR thresholds determined behavioral thresholds in patients with severe hearing losses were better than for mild hearing losses. The use of a chirp-ABR testing ensures higher sensitivity and accuracy than that of auditory stead-state evoked response (ASSR) for measuring frequency-specific thresholds in young children.

Performance of two hearing screening protocols in NICU in Shanghai.

OBJECTIVE: To study the sensitivity and specificity of targeted neonatal hearing screening for the single-session distortion product otoacoustic emissions (DPOAE) technique and the combined DPOAE/automated auditory brain-stem response (AABR) technique. METHODS: 3000 high-risk newborns were studied at Children's Hospital of Fudan University. They were required to take two different screening procedures separately. The first procedure consisted of DPOAE alone and the second consisted of DPOAE combined with the AABR. Based upon the etiology in high-risk babies, they were divided into four groups. In group I there were 670 very-low-birth-weight (VLBW) newborns (1340 ears), and in group II there were 890 preterm babies (1780 ears). 850 babies (1700 ears) suffered from hyperbilirubinemia in group III, whereas 790 babies (1580 ears) with asphyxia were in group IV. The babies in groups II, III, and IV came from the neonatal intensive-care unit (NICU) of our hospital. The study protocols consisted of the DPOAE alone and DPOAE combined with AABR hearing screening at an age of less than 1 month, and a diagnostic stage at the age of 2 months. RESULTS: With single-session DPOAE screening, the referral rate (8% of the NICU babies), the false-positive rate (4.96%) and the false-negative rate (0.8%) were higher. The different etiologies in NICU babies had significantly different referral rates (F-test, p<0.01). A 4.46% referral rate of hyperbilirubinemi babies was much lower. The combined DPOAE/AABR screening technique revealed a referral rate of 5.03%, a false-positive rate of 2% and a false-negative rate of 0.06%. The false-positive rate was well below the suggested 3% of the American Academy of Pediatric. Comparisons of the referral rate, false-positive rate and false-negative rate of two hearing screening protocols (DPOAE alone and combined DPOAE/AABR) revealed significant differences (t-test, p<0.05, p<0.01, p<0.01). 91 infants (3.03% of the NICU babies) who failed the combined DPOAE/AABR screening were confirmed on hearing impairment. Of 22 babies who passed DPOAE screening but failed the AABR screening had a severe to profound hearing loss based on classic ABR. These patients (24% of the NICU babies with hearing losses) with hyperbilirubinemia and asphyxia problems at newborn stage were diagnosed as auditory neuropathy based on evaluations of DPOAE screening passed, abnormal ABR and absent middle-ear muscle reflexes (MMR). CONCLUSION: Our study demonstrates the use of a combination of DPOAE and AABR testing ensures high sensitivity and acceptable specificity, and predict the AN profile in NICU babies. Our efforts identified 22 NICU babies with auditory neuropathy who hopefully will benefit from early remediation of their hearing deficit.