Explore the effect of LLY-283 on the ototoxicity of auditory cells caused by cisplatin: A bioinformatic analysis based on RNA-seq.

BACKGROUND: Cisplatin is a commonly used chemotherapeutic drug in clinics, and long-term application will lead to hearing impairment. LLY-283, an inhibitor of PRMT5, has not been reported in deafness. Our study aimed to explore the mechanism of LLY-283 in hearing impairment. MATERIALS AND METHODS: First, we performed RNA-seq (cisplatin in the experimental group and DMSO in the control group) to obtain the biological processes mainly involved in differentially expressed genes (DEGs). CCK-8 and LDH experiments were used to observe the effect of LLY-283 on cisplatin-induced auditory cell injury. ROS experiment was used to monitor the impact of LLY-283 on oxidative damage of auditory cells. Effect of LLY-283 on apoptosis of auditory cells detected by TUNEL experiment. PCR and Western blotting were used to detect the expression of genes and proteins related to auditory cell apoptosis in LLY-283 cells. Meanwhile, we explored the effect of LLY-283 on the expression of PRMT5 in cisplatin-induced hearing impaired cells at RNA and protein levels. RESULTS: Biological process analysis showed that DEGs were mainly enriched in the apoptotic process involved in morphogenesis (-Log10 P = 3.71). CCK-8 and LDH experiments confirmed that LLY-283 could save cisplatin-induced auditory cell injury. ROS experiments confirmed that LLY-283 could rescue cisplatin-induced oxidative damage to auditory cells. TUNEL experiments confirmed that LLY-283 could protect cisplatin-induced apoptosis of auditory cells. Meanwhile, LLY-283 could inhibit the expression of PRMT5 in auditory cells induced by cisplatin. CONCLUSION: LLY-283 can rescue cisplatin-induced auditory cell apoptosis injury. LLY-283 can inhibit the increase in PRMT5 expression induced by cisplatin.

Web-based transcriptome analysis determines a sixteen-gene signature and associated drugs on hearing loss patients: A bioinformatics approach.

BACKGROUND: Hearing loss is becoming more and more general. It may occur at all age and affect the language learning ability of children and trigger serious social problems. METHODS: The hearing loss differentially expressed genes (HL-DEGs) were recognized through a comparison with healthy subjects. The Gene Ontology (GO) analysis was executed by DAVID. The reactome analysis of HL-DEGs was performed by Clue-GO. Next, we used STRING, an online website, to identify crucial protein-protein interactions among HL-DEGs. Cytoscape software was employed to construct a protein-protein interaction network. MCODE, a plug-in of the Cytoscape software, was used for module analysis. Finally, we used DGIdb database to ascertain the targeted drugs for MCODE genes. RESULTS: Four hundred four HL-DEGs were identified, among which the most up-regulated 10 genes were AL008707.1, SDR42E1P5, BX005040.1, AL671883.2, MT1XP1, AC016957.1, U2AF1L5, XIST, DAAM2, and ADAMTS2, and the most down-regulated 10 genes were ALOX15, PRSS33, IL5RA, SMPD3, IGHV1-2, IGLV3-9, RHOXF1P1, CACNG6, MYOM2, and RSAD2. Through STRING database and MCODE analysis, we finally got 16 MCODE genes. These genes can be regarded as hearing loss related genes. Through biological analysis, it is found that these genes are enriched in pathways related to apoptosis such as tumor necrosis factor. Among them, MMP8, LTF, ORM2, FOLR3, and TCN1 have corresponding targeted drugs. Foremost, MCODE genes should be investigated for its usefulness as a new biomarker for diagnosis and treatment. CONCLUSION: In summary, our study produced a sixteen-gene signature and associated drugs that could be diagnosis and treatment of hearing loss patients.

Screening antiproliferative drug for breast cancer from bisbenzylisoquinoline alkaloid tetrandrine and fangchinoline derivatives by targeting BLM helicase.

BACKGROUND: The high expression of BLM (Bloom syndrome) helicase in tumors involves its strong association with cell expansion. Bisbenzylisoquinoline alkaloids own an antitumor property and have developed as candidates for anticancer drugs. This paper aimed to screen potential antiproliferative small molecules from 12 small molecules (the derivatives of bisbenzylisoquinoline alkaloids tetrandrine and fangchinoline) by targeting BLM642-1290 helicase. Then we explore the inhibitory mechanism of those small molecules on proliferation of MDA-MB-435 breast cancer cells. METHODS: Fluorescence polarization technique was used to screen small molecules which inhibited the DNA binding and unwinding of BLM642-1290 helicase. The effects of positive small molecules on the ATPase and conformation of BLM642-1290 helicase were studied by the malachite green-phosphate ammonium molybdate colorimetry and ultraviolet spectral scanning, respectively. The effects of positive small molecules on growth of MDA-MB-435 cells were studied by MTT method, colony formation and cell counting method. The mRNA and protein levels of BLM helicase in the MDA-MB-435 cells after positive small molecule treatments were examined by RT-PCR and ELISA, respectively. RESULTS: The compound HJNO (a tetrandrine derivative) was screened out which inhibited the DNA binding, unwinding and ATPase of BLM642-1290 helicase. That HJNO could bind BLM642-1290helicase to change its conformationcontribute to inhibiting the DNA binding, ATPase and DNA unwinding of BLM642-1290 helicase. In addition, HJNO showed its inhibiting the growth of MDA-MB-435 cells. The values of IC50 after drug treatments for 24 h, 48 h and 72 h were 19.9 µmol/L, 4.1 µmol/L and 10.9 µmol/L, respectively. The mRNA and protein levels of BLM helicase in MDA-MB-435 cells increased after HJNO treatment. Those showed a significant difference (P < 0.05) compared with negative control when the concentrations of HJNO were 5 µmol/L and 10 µmol/L, which might contribute to HJNO inhibiting the DNA binding, ATPase and DNA unwinding of BLM helicase. CONCLUSION: The small molecule HJNO was screened out by targeting BLM642-1290 helicase. And it showed an inhibition on MDA-MB-435 breast cancer cells expansion.

[Duplex real-time PCR assay for rapid detection of E. coli O157 and E. coli O104 in the legume plants and its products].

OBJECTIVE: To establish a rapid assay to detect enterohemorrhagic Escherichia coli O157 (E. coli O157) and enterohemorrhagic Escherichia coli O104 (E. coli O104) in the legume plants and its products by using the duplex real-time PCR. METHODS: Based on part fragments of E. coli O157 specific gene rfbE and E. coli O104 specific gene wzy, primers and Taq-Man probes were designed. Results DNA (Q157): DNA (O104) = 1 : 3, DNA 2 µL, Master Mix 12.5 µL, primers (10 µmol/L) 1 µL,Taq-Man probes( 10 µmol/L) 0.5 µL, ddH2O 25 µL. The two pathogens could be detected in one reaction system, the lowest limit of detection sensitivity was 10(2) CFU/mL(g). CONCLUSION: This method is rapid and convenient, with good specificity, high sensitivity. The duplex real-time PCR assay could provide a cost-effective and convenient method for detect foodborne pathogenic bacteria.

Synergistic effect of interfacial lattice Ag(+) and Ag(0) clusters in enhancing the photocatalytic performance of TiO2.

An interfacial lattice Ag(+) doped on TiO2 (Ag(+)/TiO2) was prepared by eluting Ag(0) clusters from a hydrothermally prepared Ag(0)/Ag(+)/TiO2 composite. An Ag(+)/TiO2@Ag(0) composite photocatalyst was subsequently obtained via a secondary Ag(0) clusters loading process to the Ag(+)/TiO2. The photocatalytic activity of Ag(+)/TiO2@Ag(0) was greatly improved compared to Ag(0)/Ag(+)/TiO2 and Ag(+)/TiO2. X-ray photoelectron spectroscopy (XPS) and cyclic voltammetry (CV) testing verified that Ag(+) ions occur as an interfacial lattice Ag(+) species in the composites. The enhancement effect of the interfacial lattice Ag(+) species is exhibited by the newly-formed Ag(+)/TiO2@Ag(0) as the interfacial lattice Ag(+) is fully exposed but not overlapped with the re-loaded Ag(0) clusters. The interfacial lattice Ag(+) ions and Ag(0) clusters are both responsible for the photocatalytic performance improvement of the catalyst, in either the photocatalytic degradation of methyl orange or photocurrent measurement.

Effects of Angle of Epiglottis on Aerodynamic and Acoustic Parameters in Excised Canine Larynges.

OBJECTIVES: The aim of this study is to explore the effects of the angle of epiglottis (Aepi) on phonation and resonance in excised canine larynges. METHODS: The anatomic Aepi was measured for 14 excised canine larynges as a control. Then, the Aepis were manually adjusted to 60° and 90° in each larynx. Aerodynamic and acoustic parameters, including mean flow rate, sound pressure level, jitter, shimmer, fundamental frequency (F0), and formants (F1'-F4'), were measured with a subglottal pressure of 1.5 kPa. Simple linear regression analysis between acoustic and aerodynamic parameters and the Aepi of the control was performed, and an analysis of variance comparing the acoustic and aerodynamic parameters of the three treatments was carried out. RESULTS: The results of the study are as follows: (1) the larynges with larger anatomic Aepi had significantly lower jitter, shimmer, formant 1, and formant 2; (2) phonation threshold flow was significantly different for the three treatments; and (3) mean flow rate and sound pressure level were significantly different between the 60° and the 90° treatments of the 14 larynges. CONCLUSIONS: The Aepi was proposed for the first time in this study. The Aepi plays an important role in phonation and resonance of excised canine larynges.

Establishment and Analysis of False Vocal Folds Hypertrophy Model in Excised Canine Larynges.

OBJECTIVE: This study aimed to investigate the role of false vocal folds (FVFs) medialization in phonation and the acoustic impact of ventricular hypertrophy by establishing an FVF hypertrophy model. STUDY DESIGN: A prospective in vitro experiment was carried out. SETTING: The study was carried out using a pseudolung platform with high-speed camera in a soundproof room. MATERIALS AND METHODS: Control, degree I, and degree II FVFs hypertrophy were simulated in 10 excised larynges via fructose injection of 0.1 mL for degree I and 0.25 mL for degree II. Mean flow rate (MFR), fundamental frequencies (F0), formants, and sound pressure level were measured with a subglottal pressure of 1.5 kPa and 2.5 kPa, respectively. RESULTS: When the subglottal pressure was controlled at both at 1.5 kPa and at 2.5 kPa, the degree of FVF hypertrophy significantly influenced the distribution of the formants, F0, and MFR in excised canine larynges. Increasing the degree of hypertrophy was associated with a decrease in F0 and an increase in MFR. In degree II FVF hypertrophy models, the sound pressure level and the first formant were significantly higher (P < 0.05) than in normal models. CONCLUSION: Hypertrophy of the FVFs has a significant influence on the distribution of sound energy and is associated with changes in sound quality.

[Significance of certain experiments relevant to airflow parameters in assessment of voice function].

OBJECTIVE: To investigate the role of airflow parameters of some specific examinations in voice function assessment. METHODS: The s/z ratio, pulmonary function and phonatory aerodynamic parameters were measured in subjects with benign vocal fold lesions and with normal voice. The effect of treatment in subjects with benign vocal fold lesions was evaluated with the phonatory aerodynamic parameters. RESULTS: The value of s/z ratio in the disease group was higher than that in the normal group (P<0.05). The value of PEF was significantly different between the disease group and the normal group for male (P<0.05). MFR, MPT, PTF, SGP, PTP, VE were significantly different between the disease group and the normal group (P<0.05). MFR, MPT, PTF, SGP, PTP of the disease group after surgery for both sex were significantly different from before surgery (P<0.05). The disease group was subdivided into two groups through stroboscopic examination before and one month after surgery: the worse group (with some functional laryngeal abnormality, or organic abnormality except benign vocal fold lesion) and the better group. PTF, PTP, SGP, VE were significantly different between the worse group and the normal voice group. There was almost no significant difference for aerodynamic parameters between the better group and the normal voice group (P>0.05). There was no significant difference between the worse group after 8 weeks'voice training and the normal voice group (P>0.05). CONCLUSIONS: s/z ratio, aerodynamic parameters (MFR, MPT, SGP, PTF, PTP, VE) are valuable for the diagnosis and assessment of the voice disorders. Aerodynamic parameters are sensitive to the change of glottal function during the treatment. Voice training can increase the glottal function of patients after laryngeal microsurgery.