Nomo1 deficiency causes autism-like behavior in zebrafish.

Patients with neuropsychiatric disorders often exhibit a combination of clinical symptoms such as autism, epilepsy, or schizophrenia, complicating diagnosis and development of therapeutic strategies. Functional studies of novel genes associated with co-morbidities can provide clues to understand the pathogenic mechanisms and interventions. NOMO1 is one of the candidate genes located at 16p13.11, a hotspot of neuropsychiatric diseases. Here, we generate nomo1-/- zebrafish to get further insight into the function of NOMO1. Nomo1 mutants show abnormal brain and neuronal development and activation of apoptosis and inflammation-related pathways in the brain. Adult Nomo1-deficient zebrafish exhibit multiple neuropsychiatric behaviors such as hyperactive locomotor activity, social deficits, and repetitive stereotypic behaviors. The Habenular nucleus and the pineal gland in the telencephalon are affected, and the melatonin level of nomo1-/- is reduced. Melatonin treatment restores locomotor activity, reduces repetitive stereotypic behaviors, and rescues the noninfectious brain inflammatory responses caused by nomo1 deficiency. These results suggest melatonin supplementation as a potential therapeutic regimen for neuropsychiatric disorders caused by NOMO1 deficiency.

Effects of two chd2-knockout strains on the morphology and behavior in zebrafish.

The chromodomain helicase DNA binding domain 2 (CHD2) gene is an ATPase and a member of the SNF2-like family of helicase-related enzymes. CHD2 plays critical roles in human brain development and function, and homozygous mutation of Chd2 in mice results in perinatal lethality. To further elucidate the effects of chd2, we used CRISPR/Cas9 to create two chd2-knockout strains (fdu901, 11,979-11982delGGGT, and fdu902, 27350delG) in zebrafish. We found that the deformity and mortality rates of fdu901 and fdu902 were higher than those of the wild type. Developmental delay was more obvious and embryo mortality was higher in fdu901 than in fdu902. However, the embryo deformity rate in fdu902 was higher than that in fdu901. Although there were no significant differences in behavior between the two knockout zebrafish and wild-type zebrafish at 7 days post fertilization (dpf), fdu901 and fdu902 zebrafish showed different alterations. The excitability of fdu902 was higher than that of fdu901. Overall, our data demonstrate that two homozygous chd2 knockout mutations were survivable and could be stably inherited and that fdu901 and fdu902 zebrafish differed in behavior and morphology. These two models might be good tools for understanding the functions of the different domains of chd2.

Key Factors Influencing the Operationalization and Effectiveness of Telemedicine Services in Henan Province, China: Cross-Sectional Analysis.

BACKGROUND: Telemedicine has demonstrated its potential in alleviating the unbalanced distribution of medical resources across different regions. Henan, a province in China with a population of approximately 100 million, is especially affected by a health care divide. The province has taken a proactive step by establishing a regional collaborative platform for telemedicine services provided by top-tier provincial hospitals. OBJECTIVE: We aim to identify the key factors that influence the current operationalization and effectiveness of telemedicine services in Henan province. The insights gained from this study will serve as valuable references for enhancing the efficient operation of telemedicine platforms in low- and middle-income regions. METHODS: We analyzed service reports from the performance management system of telemedicine services in Henan province throughout 2020. Using descriptive statistics and graphical methods, we examined key influencing factors, such as management competency; device configuration; and hospital capability, capacity, and service efficacy, across hospitals at 2 different tiers. In addition, we used generalized linear models and multiple linear regression models to identify key operational factors that significantly affect the service volume and efficacy of 2 major telemedicine services, namely teleconsultation and tele-education. RESULTS: Among the 89 tier 3 hospitals and 97 tier 2 hospitals connected to the collaborative telemedicine platform, 65 (73%) and 55 (57%), respectively, have established standardized management procedures for telemedicine services. As the primary delivery method for telemedicine services, 90% (80/89) of the tier 3 hospitals and 94% (91/97) of the tier 2 hospitals host videoconferencing consultations through professional hardware terminals rather than generic computers. Teleconsultation is the dominant service type, with an average annual service volume per institution of 173 (IQR 37-372) and 60 (IQR 14-271) teleconsultations for tier 3 and tier 2 hospitals, respectively. Key factors influencing the service volume at each hospital include available funding, management competency, the number of connected upper tiers, and the number of professional staff. After receiving teleconsultations from tier 3 (65/89, 73%) and tier 2 (61/97, 63%) hospitals, patients reported significant improvements in their medical conditions. In addition, we observed that service efficacy is positively influenced by management competency, financial incentives, the number of connected upper or lower tiers, and the involvement of participating medical professionals. CONCLUSIONS: Telemedicine has become increasingly popular in Henan province, with a notable focus on teleconsultation and tele-education services. Despite its popularity, many medical institutions, especially tier 2 hospitals, face challenges related to management competency. In addition to enhancing the effectiveness of existing telemedicine services, health care decision-makers in Henan province and other low- and middle-income regions should consider expanding the service categories, such as including remote emergency care and telesurgery, which have promise in addressing crucial health care needs in these regions.

ES Sampler, a minimally invasive endometrial sampling tool that obtains specimens suitable for both cytological and histological analysis.

AIM: Conventional endometrial examination by dilatation and curettage (D&C) is not accepted by many patients because it is associated with pain and risk of injury and typically requires anesthesia and hospitalization. While several less invasive endometrial screening tools have been developed, their diagnostic value is generally inferior to D&C. Therefore, the purpose of this study was to evaluate the effectiveness of a new, minimally invasive device, called the ES Sampler, for outpatient endometrial screening. METHODS: This was a single-blind study of 96 patients (age: 36.8 ± 8.1 years) who attended Peking Union Medical College Hospital from March 2015 to August 2016. Specimens were collected from each participant using the ES Sampler, followed by traditional D&C by hysteroscopy, and evaluated by histology and/or cytology. The sampling adequacy, sensitivity, specificity, positive and negative predictive value, and diagnostic accuracy were compared, and patient acceptability was assessed. RESULTS: Compared to traditional D&C, the ES Sampler exhibited 99.0% sampling adequacy, and the combined (histology and cytology) results demonstrated 88.9% sensitivity, 95.6% specificity, 88.9% positive predictive value, 95.6% negative predictive value, and 93.7% accuracy. Moreover, the majority of study participants reported mild or no pain associated with the ES Sampler, and blood loss was minimal. CONCLUSIONS: Our findings suggest that the minimally invasive ES Sampler is a reliable and accurate endometrial screening tool that is easily accepted by patients. The ES Sampler could be useful for screening high-risk patients who may need further, more invasive examination, thereby conserving medical resources and minimizing patient discomfort.

Deletion of c16orf45 in zebrafish results in a low fertilization rate and increased thigmotaxis.

c16orf45 is located at 16p13.11, an important locus related to neurodevelopmental diseases. Clinical studies have demonstrated that c16orf45 is associated with various neurodevelopmental diseases. To further elucidate the effect of c16orf45 on neural development, we constructed a zebrafish model with a stably inherited c16orf45 deletion via CRISPR/Cas9 technology. We found that deletion of c16orf45 significantly reduced the zebrafish fertilization rate, and both females and males showed reduced fertility. Meanwhile, the homozygous c16orf45 knockout zebrafish showed a developmental delay at 24 hr postfertilization (hpf). However, morphological changes were not apparent after 2 days postfertilization (dpf). Notably, the results of behavioral experiments revealed increased thigmotaxis in c16orf45-/- zebrafish at 2 months. In conclusion, these findings demonstrate that c16orf45 plays an important role in nervous system and reproductive system.

Behavioural responses of zebrafish larvae to acute ethosuximide exposure.

The anticonvulsant drug ethosuximide has shown diverse anxiety-related activity in rodents, but research in zebrafish is limited. To evaluate the effects of acute ethosuximide exposure on locomotor activity and anxiety-related thigmotaxis behaviours of zebrafish larvae, the activity of AB strain zebrafish larvae at 5 and 7 days postfertilization (dpf) was analysed under normal constant illumination and stressful light-dark transitions. Under constant illumination, ethosuximide at concentrations of 2, 5 and 10 mmol/l increased the distances travelled and intensified locomotor responses to a novel environment. In addition, 40 mmol/l ethosuximide decreased the travel distance and attenuated the locomotor response to darkness. The effects were age related. Under constant illumination, ethosuximide at 40 mmol/l reduced thigmotaxis behaviours in larvae at both ages. Under the light-dark transition, 5 mmol/l ethosuximide reduced thigmotaxis behaviours in 7-dpf larvae. We concluded that under constant lighting, ethosuximide at low concentrations (2, 5 and 10 mmol/l) stimulated the locomotor activity of zebrafish larvae, whereas a high concentration (40 mmol/l) inhibited the activity. Ethosuximide at a low concentration (5 mmol/l) showed anxiolytic effects during the stressful light-dark transition in 7-dpf larvae. The effects of ethosuximide were age and concentration related.

Effects of picrotoxin on zebrafish larvae behaviors: A comparison study with PTZ.

Larval zebrafish (Danio rerio) have been suggested as a high-throughput experimental animal model for epilepsy-related genetic and developmental studies. The behavioral manifestations in response to the seizure-inducing drugs picrotoxin (PTX) (1, 5, 25, 125, or 625µM) or pentylenetetrazole (PTZ) (1, 2, 4, 8, or 16mM) under light-dark conditions were studied using zebrafish larvae at 5days post-fertilization (dpf). Two behavioral parameters, locomotor activity and thigmotaxis behavior, were analyzed. We conclude that high concentrations of PTX treatment increased locomotion and thigmotaxis in 5 dpf zebrafish larvae under continuous illumination and the locomotion of PTX-treated zebrafish was decreased under the dark condition. High concentrations of PTX treatment also increased thigmotaxis (an indicator of increased anxiety levels) in zebrafish larvae under both continuous illumination and dark condition. PTZ treatment increased the locomotion of 5 dpf zebrafish larvae under continuous illumination. However, 2mM PTZ decreased locomotion, and high concentrations of PTZ decreased the locomotion of larvae under dark conditions. High concentrations of PTZ treatment also increased thigmotaxis in the zebrafish larvae under both continuous illumination and dark condition. Compared with PTZ, PTX leads to higher levels of movement under light conditions and lower levels of movement under dark condition. However, the level of thigmotaxis in the zebrafish larvae was similar between the two drug treatments.

Simultaneous Rapid Detection of Multiple Physicochemical Properties of Jet Fuel Using Near-Infrared Spectroscopy.

Jet fuel is the primary fuel used in the aviation industry, and its quality has a direct impact on the safety and operational efficiency of aircraft. The accurate quantitative detection and analysis of various physicochemical property indicators are important for improving and ensuring the quality of jet fuel in the domestic market. This study used near-infrared (NIR) spectroscopy to establish a suitable model for the simultaneous and rapid detection of multiple physicochemical properties in jet fuel. Using more than 40 different sources of jet fuel, a rapid detection model was established by optimizing the spectral processing methods. The measurement models were separately built using the partial least-squares (PLS) and orthogonal PLS algorithms, and the model parameters were optimized. The results show that after the Savitzky-Golay second derivative preprocessing, the PLS model built using the feature spectra selected by the uninformative variable elimination wavelength algorithm achieved the best measurement performance. Compared with the PLS model without preprocessing, the range of the resulting accuracy improvement was at least 15.01%. Under the optimal model parameters, the calibration set regression coefficient (Rc2) of the 11 jet fuel property index models ranged from 0.9102 to 0.9763, with the root-mean-square error of calibration values up to 0.8468 °C (for flash points). The regression coefficient (Rp2) of the validation set ranged from 0.8239 to 0.9557, with the root-mean-square error of prediction values up to 1.1354 °C (for flash points). The ratios of prediction to deviation (RPD) values were all in the range of 1.9-3.0, indicating high accuracy and reliability of the model. The rapid NIR analysis method established in this study enables the simultaneous and rapid detection of multiple physicochemical properties of jet fuel, thereby providing effective technical support for ensuring the quality of jet fuel in the market.

Accurate identification of methanol and ethanol gasoline types and rapid detection of the alcohol content using effective chemical information.

Methanol and ethanol gasoline are two emerging clean energy sources with different characteristics. To achieve the qualitative identification and quantitative analysis of the alcohols present in methanol and ethanol gasoline, effective chemical information (ECI) models based on the characteristic spectral bands of the near-infrared (NIR) spectra of the methanol and ethanol molecules were developed using the partial least squares discriminant analysis (PLS-DA) and partial least squares (PLS) algorithms. The ECI model was further compared with models built from the full wavenumber (Full) spectra, variable importance in projection (VIP) spectra, and Monte Carlo uninformative variable elimination (MC-UVE) spectra to determine the predictive performance of ECI model. Among the various qualitative identification models, it was found that the ECI-PLS-DA model, which is built using the differences in molecular chemical information between methanol and ethanol, exhibited sensitivity, specificity and accuracy values of 100%. The ECI-PLS-DA model accurately identified methanol gasoline and ethanol gasoline with different contents. In the quantitative analysis model for methanol gasoline, the methanol gasoline and ethanol gasoline ECI-PLS models exhibited the smallest root mean squared error of predictions (RMSEP) of 0.18 and 0.21% (v/v), respectively, compared to the other models. Meanwhile, the F-test and T-test results revealed that the NIR method employing the ECI-PLS model showed no significant difference compared to the standard method. Compared with other spectral models examined herein, the ECI model demonstrated the highest recognition success and determination accuracy. This study therefore established a highly accurate and rapid determination model for the qualitative identification and quantitative analysis based on chemical structures. It is expected that this model could be extended to the NIR analysis of other physicochemical properties of fuel.

Overexpression of miR-124-3p affects zebrafish inner ear development and hearing function via downregulation of EYA1 gene expression.

The EYA1 gene is essential for normal inner ear development and affects the development and function of the inner ear in a dose-dependent manner. However, the mechanisms regulating EYA1 gene expression are not well understood. Recently, miRNAs have become recognized as important regulators of gene expression. In this study, we identified miR-124-3p through a microRNA (miRNA) target prediction website and found that miR-124-3p and its target site in the EYA1 3' untranslated region (3'UTR) are conserved in most vertebrates. Both in vivo and in vitro, the interaction of miR-124-3p with the EYA1 3'UTR exerts a negative regulatory effect. Microinjection of agomiR-124-3p into zebrafish embryos resulted in a phenotype of reduced auricular area, suggesting inner ear dysplasia. In addition, injection of agomiR-124-3p or antagomiR-124-3p caused abnormal hearing function in zebrafish. In conclusion, our results suggest that miR-124-3p can affect inner ear development and hearing function in zebrafish by regulating EYA1.

Identification of Scd5 as a functional regulator of visceral fat deposition and distribution.

Ectopic deposition of visceral adipose tissue (VAT) in abdomen is usually accompanied with systematic chaos of energy metabolism, a higher risk of cardiovascular diseases and type II diabetes. Here, we identified a previously unexplored gene Scd5 as a master regulator of fat distribution, which alone plays a significant role in determining the VAT accumulation. Firstly, zebrafish scd5 had the highest homology with human SCD5 compared to other SCDs in mouse and rat. We then observed that scd5-homozygous mutant zebrafish displayed a puffy, short and rounded apple-shaped figure. Whole-mount micro-CT scan showed that excessive VAT deposition and short spine are responsible for the abnormal body ratio. And the supplementation of ω3-polyunsaturated fatty acid (ω3-PUFA) in dietary significantly decreased VAT accumulation in scd5 -/- zebrafish. Lastly, transcriptional analyses revealed that the Wnt, PPAR, C/EBP, and fat synthesis signaling pathways are significantly affected in the VAT of scd5 -/- mutant and restored by ω3-PUFA.

Rrn3 gene knockout affects ethanol-induced locomotion in adult heterozygous zebrafish.

Genome-wide analysis has identified the transcription factor, RRN3 (or TIF-1A), on human chromosome 16p13.11 as a candidate gene associated with mental disorders. Both genetic and biochemical experiments have demonstrated that RRN3 plays a major role in the transcriptional regulation of ribosomal DNA and cell growth. Previous research has suggested that loss of RRN3 from mature neurons reproduces the chronic nature of neurodegenerative processes. Here, we report the first generation and characterization of rrn3 mutant zebrafish in larval and adult stages using the CRISPR/Cas9 genome editing technique. Homozygous knockout zebrafish exhibited morphological changes, such as pericardial oedema and deformed heads, and died at the larval stage of embryonic development. Behaviourally, the locomotion and shoaling behaviour of adult rrn3+/- zebrafish was not significantly different compared with rrn3+/+ zebrafish. Notably, rrn3+/- zebrafish demonstrated abnormal locomotor activity in response to ethanol. We found decreased norepinephrine expression in the brains of rrn3+/- zebrafish when treated with ethanol. In summary, our results indicated that rrn3 was closely associated with early embryonic development in zebrafish. Furthermore, behavioural and neurochemical research revealed the importance of genetic differences in drug sensitivity. The results suggest that caution should be taken when treating RRN3 heterozygous patients.

Deficiency of nde1 in zebrafish induces brain inflammatory responses and autism-like behavior.

The cytoskeletal protein NDE1 plays an important role in chromosome segregation, neural precursor differentiation, and neuronal migration. Clinical studies have shown that NDE1 deficiency is associated with several neuropsychiatric disorders including autism. Here, we generated nde1 homologous deficiency zebrafish (nde1 -/- ) to elucidate the cellular molecular mechanisms behind it. nde1 -/- exhibit increased neurological apoptotic responses at early infancy, enlarged ventricles, and shrank valvula cerebelli in adult brain tissue. Behavioral analysis revealed that nde1 -/- displayed autism-like behavior traits such as increased locomotor activity and repetitive stereotype behaviors and impaired social and kin recognition behaviors. Furthermore, nde1 mRNA injection rescued apoptosis in early development, and minocycline treatment rescued impaired social behavior and overactive motor activity by inhibiting inflammatory cytokines. In this study, we revealed that nde1 homozygous deletion leads to abnormal neurological development with autism-related behavioral phenotypes and that inflammatory responses in the brain are an important molecular basis behind it.

The tissue-specificity associated region and motif of an emx2 downstream enhancer CNE2.04 in zebrafish.

BACKGROUND: Expression level of EMX2 plays an important role in the development of nervous system and cancers. CNE2.04, a conserved enhancer downstream of emx2, drives fluorescent protein expression in the similar pattern of emx2. METHODS: CNE2.04 truncated or motif-mutated transgenic reporter plasmids were constructed and injected into the zebrafish fertilized egg with Tol2 mRNA at the unicellular stage of zebrafish eggs. The green fluorescence expression patterns were observed at 24, 48, and 72 hpf, and the fluorescence rates of different tissues were counted at 48 hpf. RESULTS: Compared to CNE2.04, CNE2.04-R400 had comparable enhancer activity, while the tissue specificity of CNE2.04-L400 was obviously changed. Motif CCCCTC mutation obviously changed the enhancer activity, while motif CCGCTC mutations also changed it. CONCLUSION: Due to their correlation with tissue specificity, CNE2.04-R400 is associated with the tissue-specificity of CNE2.04, and motif CCCCTC plays an important role in the enhancer activity of CNE2.04.

Comparative sequence analysis and functional validation identified a retina-specific enhancer around zic5 and zic2a.

The spatiotemporal development of vertebrate retina is regulated by a variety of genes. The zinc finger transcription factors zic5 and zic2a are located close to each other in the chromosome. They have similar expression patterns, and both play important roles in the development of the retina and nervous system. Here, we used ECR browser and gfp fluorescence report experiment to identify a 290bp enhancer sequence ECR3, which is located at 3 kb upstream of zic5 and 10 kb downstream of zic2a, and it can drive the specific expression of gfp in the retina. pT2KXIGQ-ECR3 was used to construct a transgenic zebrafish line Tg(ECR3-290: gfp) which first exhibits specific green fluorescence in the whole retina area at 24hpf. Then the expression region was gradually limited to ganglion cell layer (GCL) and lasted through adulthood. This expression pattern is highly consistent with the zic5 and zic2a at retina. These results indicate that the 290bp enhancer might be an important element to regulate the expression of zic5 and zic2a genes in ganglion cells, and this transgenic line is an important tool for studying the development of retina.

Phosphoproteomics Reveals Key Regulatory Kinases and Modulated Pathways Associated with Ovarian Cancer Tumors.

BACKGROUND: Ovarian cancer (OC) is the seventh most common cancer worldwide for women. However, there are no sufficient diagnostic methods and few treatment options available due to poor understanding of its pathogenic mechanisms. METHODS: To comprehensively analyze the phosphoproteomic characterization for OC, we took advantage of a quantitative global phosphoproteomics method, titanium(IV) immobilized metal affinity chromatography (Ti4+-IMAC) coupled to nanoscale liquid chromatography and quadrupole time-of-flight tandem mass spectrometry (nanoLC/Q-TOF-MS/MS) on ovarian tissue samples obtained from five OC patients and five matched controls. RESULTS: A total of 722 phosphorylated sites corresponding to 534 proteins were significantly different (fold change ≥ 2, p < 0.01) between OC patients and the controls. Among them, 83 transcription factors mainly consisted of transcription cofactors, zf-C2H2, and chromatin remodeling factors and 29 kinases were included. Further functional analysis suggested significantly biological processes were highly enriched and involved in the pathogenesis of OC, especially fructose and mannose metabolism. Moreover, the regulatory roles of modulated pathways, including MAPK, ErbB, and GnRH signaling pathways were also identified as critical processes involved in OC. The results here highlighted key phosphorylated proteins, particularly kinases, and the corresponding cancer-related metabolic and signal pathways that played important roles in the development of OC. Additionally, the expression levels of two kinases, phosphorylated CDK (T14) and phosphorylated PRKCQ (S695), were validated by Western blot analysis in the other group of ovarian tissue samples. CONCLUSION: Altogether, our data not only provided novel insights into the potential biomarkers and therapy options for OC but also extended our knowledge on its pathophysiological mechanism.

[Effects of Kechuanluo oral liquid on eosinophil apoptosis and its regulation factors in lung tissues of asthmatic mice].

OBJECTIVE: To explore the mechanism of Kechuanluo oral liquid, a compound Chinese herbal medicine, in inhibiting allergic airway inflammation by observing the effects of Kechuanluo on eosinophil (EOS) apoptosis and its regulation factors in asthmatic mice. METHODS: Fifty-six BALB/c mice were randomly divided into normal control group (n=16), untreated group (n=16), Western medicine group (n=12) and Kechuanluo group (n=12). Except for the mice in normal control group, asthma was induced in BALB/c mice by using ovalbumin (OVA) and potassium aluminium sulfate. The mice were intragastrically administered with normal saline, Kechuanluo (30 ml/kg daily) and prednisolone tablets (10 mg/kg daily) respectively for two weeks. At 0 hour, the 3rd, 7th and 14th day after the end of OVA sensitization, the EOSs of lung tissues were counted by improved-Giemsa staining method; immunohistochemical method and image analysis were used to detect the expressions of Fas, FasL and Bcl-XL in the EOSs in the four groups; and apoptotic rates of the EOSs in the lung tissues of asthmatic mice were detected by terminal deoxynucleotidyl transferase-mediated biotin-dUTP nick end labeling (TUNEL) technique. RESULTS: Compared with the untreated group, airway inflammations of the mice in the Kechualuo group and Western medicine group were lessened and the EOS counts decreased on the 3rd, 7th and 14th day after the last OVA sensitization (P<0.05). On the 3rd day, the EOSs apoptotic rates, the expression areas of Fas in the EOSs and FasL in the lung tissues were significantly higher in the Kechuanluo group than those in the untreated group (P<0.01), furthermore, the EOS apoptotic rate reached the peak level. Inversely, the EOS count and the expression area of Bcl-XL in the EOSs were obviously lower in the Kechuanluo group (P<0.05). On the 7th and 14th day, the expression areas of Fas and Bcl-XL in the EOSs were significantly decreased in the Kechuanluo group (P<0.01), and on the 14th day, the EOS apoptotic rate and the expression area of FasL in the lung tissues were obviously lower either. The effects exhibited in the Western medicine group were similar to those in the Kechuanluo group. CONCLUSION: There is airway inflammation with eosinophilic infiltration in asthmatic mice, accompanied with suppressed apoptosis and delayed apoptosis. Kechuanluo can induce and accelerate EOS apoptosis in early inflammation by inhibiting eosinophilic inflammation, improving Fas expression in EOSs and FasL expression in the lung tissues, and reducing Bcl-XL expression in EOSs.

Sound shock response in larval zebrafish: A convenient and high-throughput assessment of auditory function.

Given that hearing ability can be challenged in diverse ways, it is necessary to develop an easily conducted, high-throughput method for assessing potential auditory risks. Measuring the acoustic startle response (ASR) has become a critical behavioral method in hearing research using zebrafish (Danio rerio). In this study, changes in the activity of zebrafish larvae (10 days post fertilization (dpf)) due to exposure to a sudden easily-generated broad-band noise were automatically and objectively recorded and analyzed without building sophisticated equipments. A significant increase in activity was induced by the noise stimulation and the alterations were impaired by gentamicin. In addition, a clear dose-response trend was observed between gentamicin exposure and the impaired activity, and a similar phenomenon was observed between gentamicin exposure and damage to hair cells. Our results suggested that alterations in the activity induced by a broad-band noise can potentially be used as an efficient assay for assessing hearing ability.

ARID1A gene silencing reduces the sensitivity of ovarian clear cell carcinoma to cisplatin.

In ovarian clear cell carcinoma (OCCC), the mutation rate of the AT-rich interaction domain 1A (ARID1A) gene is 46-57%. However, the effects of ARID1A gene silencing by small interfering RNA (siRNA) on the sensitivity of OCCC to cisplatin have not been investigated. Thus, this study aimed to elucidate the association between ARID1A gene silencing and drug resistance in OCCC. Three pairs of ARID1A gene siRNA fragments (siRNA-1, siRNA-2 and siRNA-3) were designed and transiently transfected into ES2 OCCC cells using RNAi Max reagent. Western blotting results demonstrated that the transfection reduced ARID1A protein expression levels, with the siRNA-3 group having the lowest levels. The IC50 value, determined using a Cell Counting kit-8 assay, was significantly increased by siRNA-3 transfection compared with that in blank control and negative control groups. The cell survival rate following treatment with 50 µM cisplatin for 48 h was significantly increased in the siRNA-3 group compared with the blank control and negative control groups. Flow cytometric analysis revealed that the apoptosis rate for cisplatin-treated cells was significantly lower in cells with siRNA-3 transfection than in those without, and the apoptosis rate in siRNA-3-transfected cells was lower than that in the negative control group. Western blot analysis showed that the expression level of AKT in cisplatin-treated cells was significantly decreased compared with that in the negative control group, and the AKT expression level in cisplatin-treated cells was significantly higher with siRNA-3 transfection than without. Therefore, the results demonstrated that ARID1A siRNA efficiently decreased ARID1A expression, which reduced cisplatin chemosensitivity and cell apoptosis in ES2 OCCC cells via the regulation of AKT expression.