Identification of miRNA expression profile in middle ear cholesteatoma using small RNA-sequencing.

BACKGROUND: The present study aims to identify the differential miRNA expression profile in middle ear cholesteatoma and explore their potential roles in its pathogenesis. METHODS: Cholesteatoma and matched normal retroauricular skin tissue samples were collected from patients diagnosed with acquired middle ear cholesteatoma. The miRNA expression profiling was performed using small RNA sequencing, which further validated by quantitative real-time PCR (qRT-PCR). Target genes of differentially expressed miRNAs in cholesteatoma were predicted. The interaction network of 5 most significantly differentially expressed miRNAs was visualized using Cytoscape. Further Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genome (KEGG) pathway enrichment analyses were processed to investigate the biological functions of miRNAs in cholesteatoma. RESULTS: The miRNA expression profile revealed 121 significantly differentially expressed miRNAs in cholesteatoma compared to normal skin tissues, with 56 upregulated and 65 downregulated. GO and KEGG pathway enrichment analyses suggested their significant roles in the pathogenesis of cholesteatoma. The interaction network of the the 2 most upregulated (hsa-miR-21-5p and hsa-miR-142-5p) and 3 most downregulated (hsa-miR-508-3p, hsa-miR-509-3p and hsa-miR-211-5p) miRNAs identified TGFBR2, MBNL1, and NFAT5 as potential key target genes in middle ear cholesteatoma. CONCLUSIONS: This study provides a comprehensive miRNA expression profile in middle ear cholesteatoma, which may aid in identifying therapeutic targets for its management.

Mifepristone protects acetaminophen induced liver injury through NRF2/GSH/GST mediated ferroptosis suppression.

Ferroptosis is a form of iron-dependent cell death that has attracted significant attention for its potential role in numerous diseases. Targeted inhibition of ferroptosis could be of potential use in treating diseases: such as drug induced liver injury (DILI). Ferroptosis can be antagonized by the xCT/GSH/GPX4, FSP1/CoQ10, DHODH/CoQ10, GCH1/BH4, and NRF2 pathways. Identifying novel anti-ferroptosis pathways will further promote our understanding of the biological nature of ferroptosis and help discover new drugs targeting ferroptosis related human diseases. In this study, we identified the clinically used drug mifepristone (RU486) as a novel ferroptosis inhibitor. Mechanistically, RU486 inhibits ferroptosis by inducing GSH synthesis pathway, which supplies GSH for glutathione-S-transferase (GST) mediated 4-HNE detoxification. Furthermore, RU486 induced RLIP76 and MRP1 export 4-HNE conjugate contributes to its anti-ferroptosis activity. Interestingly, RU486 induced GSH/GSTs/RLIP76&MRP1 anti-ferroptosis pathway acts independent of classic anti-ferroptosis systems: including xCT/GSH/GPX4, FSP1, DHODH, GCH1, SCD1 and FTH1. Moreover, NRF2 was identified to be important for RU486's anti-ferroptosis activity by inducing downstream gene expression. Importantly, in mouse model, RU486 showed strong protection effect on acetaminophen (APAP)-induced acute liver injury, evidenced by decreased ALT, AST level and histological recovery after APAP treatment. Interestingly, RU486 also decreased oxidative markers, including 4-HNE and MDA, and induced NRF2 activation as well as GSTs, MRP1 expression. Together, these data suggest NRF2/GSH/GST/RLIP76&MRP1 mediated detoxification pathway as an important independent anti-ferroptosis pathway act both in vitro and in vivo.

Effects of second language acquisition on brain functional networks at different developmental stages.

Previous studies have shown that language acquisition influences both the structure and function of the brain. However, whether the acquisition of a second language at different periods of life alters functional network organization in different ways remains unclear. Here, functional magnetic resonance imaging data from 27 English-speaking monolingual controls and 52 Spanish-English bilingual individuals, including 22 early bilinguals who began learning a second language before the age of ten and 30 late bilinguals who started learning a second language at age fourteen or later, were collected from the OpenNeuro database. Topological metrics of resting-state functional networks, including small-world attributes, network efficiency, and rich- and diverse-club regions, that characterize functional integration and segregation of the networks were computed via a graph theoretical approach. The results showed obvious increases in network efficiency in early bilinguals and late bilinguals relative to the monolingual controls; for example, the global efficiency of late bilinguals and early bilinguals was improved relative to that of monolingual controls, and the local efficiency of early bilinguals occupied an intermediate position between that of late bilinguals and monolingual controls. Obvious increases in rich-club and diverse-club functional connectivity were observed in the bilinguals relative to the monolingual controls. Three network metrics were positively correlated with Spanish proficiency test scores. These findings demonstrated that early and late acquisition of a second language had different impacts on the functional networks of the brain.

Presence of SARS-CoV-2 in middle ear fluid and characterization of otitis media with effusion in patients with COVID-19.

OBJECTIVES: This study sought to determine whether SARS-CoV-2 is present in the middle ear fluid (MEF) of patients with COVID-19 who have otitis media with effusion (OME). METHODS: A case-control study was designed to detect SARS-CoV-2 and six other common respiratory viruses (influenza A virus, influenza B virus, respiratory syncytial virus, adenovirus, human rhinovirus [HRV], and mycoplasma pneumonia) by polymerase chain reaction (PCR) in the MEF of patients with OME. Follow-up tests, including pure-tone audiometry and tympanometry, were conducted. RESULTS: In the COVID-19 group, 18 of 27 MEF specimens were PCR-positive for SARS-CoV-2, with cycle thresholds ranging from 24.9-42.2. And one patient tested positive for the HRV. In the control group, all 15 MEF specimens were PCR-negative for SARS-CoV-2, but two patients tested positive for the HRV. After treatment for OME, 11 patients (40.7%) reported complete resolution, 14 (51.9%) reported improvement, and two (7.4%) reported no change. The average improvement in hearing was 14.5 ± 8.1 dB, and the average air-bone gap decrease was 13.5 ± 9.0 dB. CONCLUSION: This study confirmed the presence of SARS-CoV-2 in the MEF of patients who were previously COVID-19-positive suggesting a possible association between COVID-19 and OME. OME should be considered a possible symptom of COVID-19.

Functional Connectome Hierarchy Distortions in Female Nurses With Occupational Burnout and Its Gene Expression Signatures.

BACKGROUND: Burnout has become a serious public health issue worldwide, particularly during the COVID-19 pandemic. Functional connectome impairments associated with occupational burnout were widely distributed, involving both low-level sensorimotor cortices and high-level association cortices. PURPOSE: To investigate whether there are hierarchical perturbations in the functional connectomes and if these perturbations are potentially influenced by genetic factors in nurses who feel "burned out." STUDY TYPE: Prospective, case control. POPULATION: Thirty-three female nurses with occupational burnout (aged 27-40, 32.42 ± 3.37) and 32 matched nurses who were not feeling burned out (aged 27-42, 32.50 ± 4.21). FIELD STRENGTH/SEQUENCE: 3.0 T, gradient-echo echo-planar imaging sequence (GE-EPI). ASSESSMENT: Gradient-based techniques were used to depict the perturbations in the multi-dimensional hierarchical structure of the macroscale connectome. Gene expression data were acquired from the Allen Human Brain Atlas. STATISTICAL TESTS: Cortex-wide multivariate analyses were used for between-group differences in gradients as well as association analyses between the hierarchy distortions and the MBI score (FDR corrected). Partial least squares, spin test and bootstrapping were utilized together to select the gene sets (FDR corrected). Gene enrichment analyses (GO, KEGG and cell-type) were further performed. Significance level: P < 0.05. RESULTS: There were significant gradient distortions, with strong between-group effects in the somatosensory network and moderate effects in the higher-order default-mode network, which were significantly correlated with the gene expression profiles (r = 0.3171). The most related genes were broadly involved in the cellular response to minerals, neuronal plasticity, and the circadian rhythm pathway (q value < 0.01). Significant enrichments were found in excitatory (r = 0.2588), inhibitory neurons (r = 0.2610), and astrocytes cells (r = 0.2633). Regions affected by burnout severity were mainly distributed in the association and visual cortices. DATA CONCLUSION: By connecting in vivo imaging to genes, cell classes, and clinical data, this study provides a framework to understand functional impairments in occupational burnout and how the microscopic genetic architecture drive macroscopic distortions. EVIDENCE LEVEL: 1 TECHNICAL EFFICACY: Stage 2.

PRMT1 inhibition promotes ferroptosis sensitivity via ACSL1 upregulation in acute myeloid leukemia.

Acute myeloid leukemia (AML) is a hematological malignancy with an alarming mortality rate. The development of novel therapeutic targets or drugs for AML is urgently needed. Ferroptosis is a form of regulated cell death driven by iron-dependent lipid peroxidation. Recently, ferroptosis has emerged as a novel method for targeting cancer, including AML. Epigenetic dysregulation is a hallmark of AML, and a growing body of evidence suggests that ferroptosis is subject to epigenetic regulation. Here, we identified protein arginine methyltransferase 1 (PRMT1) as a ferroptosis regulator in AML. The type I PRMT inhibitor GSK3368715 promoted ferroptosis sensitivity in vitro and in vivo. Moreover, PRMT1-knockout cells exhibited significantly increased sensitivity to ferroptosis, suggesting that PRMT1 is the primary target of GSK3368715 in AML. Mechanistically, both GSK3368715 and PRMT1 knockout upregulated acyl-CoA synthetase long-chain family member 1 (ACSL1), which acts as a ferroptosis promoter by increasing lipid peroxidation. Knockout ACSL1 reduced the ferroptosis sensitivity of AML cells following GSK3368715 treatment. Additionally, the GSK3368715 treatment reduced the abundance of H4R3me2a, the main histone methylation modification mediated by PRMT1, in both genome-wide and ACSL1 promoter regions. Overall, our results demonstrated a previously unknown role of the PRMT1/ACSL1 axis in ferroptosis and suggested the potential value and applications of the combination of PRMT1 inhibitor and ferroptosis inducers in AML treatment.

Association of aberrant brain network dynamics with gut microbial composition uncovers disrupted brain-gut-microbiome interactions in irritable bowel syndrome: Preliminary findings.

BACKGROUND AND PURPOSE: Growing evidence suggests that abnormalities in brain-gut-microbiome (BGM) interactions are involved in the pathogenesis of irritable bowel syndrome (IBS). Our study aimed to explore alterations in dynamic functional connectivity (DFC), the gut microbiome and the bidirectional interaction in the BGM. METHODS: Resting-state functional magnetic resonance imaging (rs-fMRI), fecal samples and clinical chacteristics were collected from 33 IBS patients and 32 healthy controls. We performed a systematic DFC analysis on rs-fMRI. The gut microbiome was analyzed by 16S rRNA gene sequencing. Associations between DFC characteristics and microbial alterations were explored. RESULTS: In the DFC analysis, four dynamic functional states were identified. IBS patients exhibited increased mean dwell and fraction time in State 4, and reduced transitions from State 3 to State 1. Aberrant temporal properties in State 4 were only evident when choosing a short window (36 s or 44 s). Decreased functional connectivity (FC) variability was found in State 1 and State 3 in IBS patients, two of which (independent component [IC]51-IC91, IC46-IC11) showed significant correlations with clinical characteristics. Additionally, we identified nine significantly differential abundances in microbial composition. We also found that IBS-related microbiota were associated with aberrant FC variability, although these exploratory results were obtained at an uncorrected threshold of significance. CONCLUSIONS: Although future studies are needed to confirm our results, the findings not only provide a new insight into the dysconnectivity hypothesis in IBS from a dynamic perspective, but also establish a possible link between DFC and the gut microbiome, which lays the foundation for future research on disrupted BGM interactions.

Impact of a Modified Restoration of Tensor Veli Palatini on Hearing and Middle Ear Function in Cleft Palate Children-a Retrospective Study.

BACKGROUND: Many children with cleft palate also exhibit hearing loss and middle ear dysfunction, which could last for years. There are still arguments on how to treat this problem. This study aimed to evaluate the impact of a modified restoration of tensor veli palatine (TVP) on hearing and middle ear function in the cleft palate children. METHODS: This retrospective study was completed using records of the cleft palate children who received surgery in Peking Union Medical College Hospital from May 2013 to December 2020. They were divided into 2 groups: Group 1: children who received cleft palate surgery without specific restoration of TVP; Group 2: children who received palate surgery with a specific TVP restoration technique. Perioperative information was collected. The conductive auditory brainstem response and the 226-Hz tympanometry before and after the cleft surgery were compared intragroup and intergroup. RESULTS: Totally 42 children were included in this study, 21 children in each group. There were no significant differences considering clinical characteristics between the 2 groups. The modified TVP restoration didn't increase operation time or complication compared with no TVP restoration. Statistically, neither the auditory brainstem response air conduction hearing thresholds nor the 226-Hz tympanometry results had significant differences between the 2 groups after the surgery. CONCLUSIONS: This modified restoration of TVP was not time-consuming and did not increase complications. The beneficial effect of the modified TVP restoration on the hearing or the middle ear function of cleft palate children was uncertain around 6 months after surgery compared with no restoration.

Intrinsic brain activity reorganization contributes to long-term compensation of higher-order hearing abilities in single-sided deafness.

Single-sided deafness (SSD) is an extreme case of partial hearing deprivation and results in a significant decline in higher-order hearing abilities, including sound localization and speech-in-noise recognition. Clinical studies have reported that patients with SSD recover from these higher-order hearing abilities to some extent over time. Neuroimaging studies have observed extensive brain functional plasticity in patients with SSD. However, studies investigating the role of plasticity in functional compensation, particularly those investigating the relationship between intrinsic brain activity alterations and higher-order hearing abilities, are still limited. In this study, we used resting-state functional MRI to investigate intrinsic brain activity, measured by the amplitude of low-frequency fluctuation (ALFF), in 19 patients with left SSD, 17 patients with right SSD, and 21 normal hearing controls (NHs). All patients with SSD had durations of deafness longer than 2 years. Decreased ALFF values in the bilateral precuneus (PCUN), lingual gyrus, and left middle frontal gyrus were observed in patients with SSD compared with the values of NHs. Longer durations of deafness were correlated with better hearing abilities, as well as higher ALFF values in the left inferior parietal lobule, the angular gyrus, the middle occipital gyrus, the bilateral PCUN, and the posterior cingulate gyrus. Moreover, we observed a generally consistent trend of correlation between ALFF values and higher-order hearing abilities in specific brain areas in patients with SSD. That is, better abilities were correlated with lower ALFF values in the frontal regions and higher ALFF values in the PCUN and surrounding parietal-occipital areas. Furthermore, mediation analysis revealed that the ALFF values in the PCUN were a significant mediator of the relationship between the duration of deafness and higher-order hearing abilities. Our study reveals significant plasticity of intrinsic brain activity in patients with SSD and suggests that reorganization of intrinsic brain activity may be one of the compensatory mechanisms that facilitate improvement in higher-order hearing abilities in these patients over time.

Cochlear Implantation in Young Mandarin-Speaking Children: One Year After First Fitting.

INTRODUCTION: Bilateral cochlear implantation has been shown to be beneficial in terms of sound localization and speech recognition in children with congenital deafness. However, little is known about the benefits of bilateral cochlear implantationin children who communicate in a tonal language such as Mandarin. This study aims to investigate the auditory perception and speech intelligibility of Mandarin-speaking children 1 year after first fitting. MATERIALS AND METHODS: Twenty-nine children aged between 11.5 and 17.9 months with severe-to-profound bilateral sensorineural hearing loss were recruited; 10 were unilaterally implanted and 19 were bilaterally implanted. A test battery was used to monitor improvements during the first year of cochlear implant use. RESULTS: Bilaterally implanted children scored better in the spatial domain of the Speech, Spatial and Qualities of hearing scale for Parents (SSQ-P) in comparison to unilaterally implanted children. Significant improvements were observed in auditory performance and speech intelligibility at 6 and 12 months after first fitting for both groups of children. DISCUSSION: Young children that speak a tonal language, such as Mandarin, can obtain significant improvements in hearing and speech abilities within the first year of cochlear implant use. Furthermore, bilateral implantation provides users with better spatial hearing in comparison to unilateral implantation.

A transformer-based multi-features fusion model for prediction of conversion in mild cognitive impairment.

Mild cognitive impairment (MCI) is usually considered the early stage of Alzheimer's disease (AD). Therefore, the accurate identification of MCI individuals with high risk in converting to AD is essential for the potential prevention and treatment of AD. Recently, the great success of deep learning has sparked interest in applying deep learning to neuroimaging field. However, deep learning techniques are prone to overfitting since available neuroimaging datasets are not sufficiently large. Therefore, we proposed a deep learning model fusing cortical features to address the issue of fusion and classification blocks. To validate the effectiveness of the proposed model, we compared seven different models on the same dataset in the literature. The results show that our proposed model outperformed the competing models in the prediction of MCI conversion with an accuracy of 83.3% in the testing dataset. Subsequently, we used deep learning to characterize the contribution of brain regions and different cortical features to MCI progression. The results revealed that the caudal anterior cingulate and pars orbitalis contributed most to the classification task, and our model pays more attention to volume features and cortical thickness features.

Aberrant functional network topology and effective connectivity in burnout syndrome.

OBJECTIVE: Recent neuroimaging studies have demonstrated that burnout is linked to specific anatomical and functional abnormalities in the brain. However, topological alterations of brain networks are not yet characterized in burnout. METHODS: Resting-state functional magnetic resonance imaging (rs-fMRI) was performed on 32 female participants with burnout and 30 matched healthy controls. Subsequently, we employed graph theoretical and network-based statistic (NBS) methods to analyze the functional connectivity. We further explored the causal influences between brain regions using the Granger Causal Analysis. Finally, partial correlation analyses were conducted between clinical scores and the altered network properties as well as connectivity metrics. RESULTS: Both the participants with burnout and healthy controls displayed a small-world organization. However, participants with burnout showed increased characteristic path length and decreased global efficiency. Corresponding local changes were mainly distributed in the visual network (2/3,66.67%). With the network-based statistic (NBS) approach, significantly decreased effective connectivities were observed mainly between the visual network and the right hippocampus. In addition, characteristic path length and nodal local efficiency of the left fusiform gyrus showed a significant negative correlation with depression severity. CONCLUSIONS: The present psychopathological findings reflect the disrupted global integration of the functional network related to the traits of participants with burnout. SIGNIFICANCE: These findings deliver novel insights from a full network perspective into the brain mechanisms of burnout.

Effect of subject state on auditory brainstem response threshold using Kalman-weighted averaging.

OBJECTIVES: This study aims to explore the impact of a subject's testing state on auditory brainstem response (ABR) thresholds using a novel ABR system (Vivosonic Integrity™), which incorporates Kalman-weighted averaging and bluetooth electrical isolation to address the limitation of conventional ABR limitation to obtain a stable result under non-sedated conditions, especially for infants and children. METHOD: Twenty-four adults (18-34 years old, 48 ears) with normal hearing were enrolled for ABR testing under three different states (lying quietly in the supine position or sleeping-lying; watching silent videos quietly in a seated position-sitting; and writing in a seated position-writing), which simulate the behaviors of young children most often encountered during non-sedated Kalman-weighted ABR testing in clinical practice. The click ABR (cABR) and tone-burst ABR (tbABR) thresholds (0.5, 1, 2, and 4 kHz) of each subject and the time taken to reach the monaural threshold for each kind of stimulus were recorded. RESULTS: (1) The cABR and tbABR thresholds were observed to increase in the following order: lying < sitting < writing. Significant threshold differences were found between any two states, except for between the sitting and lying states for the cABR and between sitting and writing for the 0.5 kHz tbABR. (2) The time required for cABR testing in the writing state was significantly longer than that in the lying and sitting states. The time required for 1 and 4 kHz tbABR testing in the lying state was significantly shorter than that in the sitting or writing state. For 2 KHz tbABR, only testing time under writing was significantly longer than that under lying. There were no significant differences in the time used for 0.5 kHz tbABR testing among different states. CONCLUSIONS: Different testing states have significant impacts on the thresholds of ABRs using Kalman-weighted averaging. A subject's state during ABR testing warrants consideration, and normal levels and correction values to estimate the hearing threshold from the ABR threshold should be determined for different testing states.

Influence of prefoldin subunit 4 on the tolerance of Kluyveromyces marxianus to lignocellulosic biomass-derived inhibitors.

BACKGROUND: Kluyveromyces marxianus is a potentially excellent host for microbial cell factories using lignocellulosic biomass, due to its thermotolerance, high growth rate, and wide substrate spectrum. However, its tolerance to inhibitors derived from lignocellulosic biomass pretreatment needs to be improved. The prefoldin complex assists the folding of cytoskeleton which relates to the stress tolerance, moreover, several subunits of prefoldin have been verified to be involved in gene expression regulation. With the presence of inhibitors, the expression of a gene coding the subunit 4 of prefoldin (KmPFD4), a possible transcription factor, was significantly changed. Therefore, KmPFD4 was selected to evaluate its functions in inhibitors tolerance. RESULTS: In this study, the disruption of the prefoldin subunit 4 gene (KmPFD4) led to increased concentration of intracellular reactive oxygen species (ROS) and disturbed the assembly of actin and tubulin in the presence of inhibitors, resulting in reduced inhibitor tolerance. Nuclear localization of KmPFD4 indicated that it could regulate gene expression. Transcriptomic analysis showed that upregulated gene expression related to ROS elimination, ATP production, and NAD+ synthesis, which is a response to the presence of inhibitors, disappeared in KmPFD4-disrupted cells. Thus, KmPFD4 impacts inhibitor tolerance by maintaining integration of the cytoskeleton and directly or indirectly affecting the expression of genes in response to inhibitors. Finally, overexpression of KmPFD4 enhanced ethanol fermentation with a 46.27% improvement in productivity in presence of the inhibitors. CONCLUSION: This study demonstrated that KmPFD4 plays a positive role in the inhibitor tolerance and can be applied for the development of inhibitor-tolerant platform strains.

Widespread plasticity of cognition-related brain networks in single-sided deafness revealed by randomized window-based dynamic functional connectivity.

As an extreme type of partial auditory deprivation, single-sided deafness (SSD) has been demonstrated to lead to extensive neural plasticity according to multimodal neuroimaging studies. Among them, resting-state functional magnetic resonance imaging (rs-fMRI) offers valuable information on functional connectivities (FCs). However, most previous SSD rs-fMRI studies assumed that the extracted FC remains stationary during the entire fMRI scan and neglected dynamic functional activities. Existing fixed window-based dynamic FC analysis also ignores dynamic functional activities under different temporal terms. Additionally, due to the cost constraints of using MRI machines, using data-driven methods for unbiased hypothesis investigations may require more effective sample data augmentation techniques. To tackle these challenges and problems together, in this study, we proposed a dynamic window with a random length and position to extract participants' dynamic characteristics under different temporal terms and to extract more information from the dataset. Then, we proposed a nodal efficiency-based correlation matrix to describe the relationships of synergism between regions as features and applied a linear support vector machine (SVM) model to learn the importance of the features, which helped to identify SSD patients and healthy controls. A total of 68 participants (including 23 with left SSD, 20 with right SSD and 25 healthy controls) were enrolled. Our proposed approach with a random window showed clear improvement compared with traditional static and fixed window-based dynamic FC by using the linear SVM model. FCs related to the frontoparietal, somatomotor, dorsal attention, limbic and default mode networks played significant roles in differentiating SSD patients from healthy controls. Additionally, FCs between the somatomotor and frontoparietal networks made the greatest contribution to the classification model. Regarding brain regions, FCs related to the superior frontal gyrus, superior parietal lobule, superior temporal gyrus, amygdala, and orbital gyrus played significant roles. These findings suggest that networks and regions related to higher-order cognitive functions showed the most significant FC alterations in SSD, which may represent a compensatory collaboration of cognitive resources in SSD.

In vitro and in vivo relaxation and anti-inflammation of natural flavonoids from Elaeagnus pungens leaf via L-type calcium channel and targeting MAPK signal pathway.

ETHNOPHARMACOLOGICAL RELEVANCE: In traditional Chinese medicine (TCM), the leaf of Elaeagnus pungens Thunb. (Family Elaeagnaceae) is a herb documented as an antiasthmatic remedy to treat the severe asthma, bronchitis and other respiratory diseases in the early material medica "Bencao Gangmu" (Ming dynasty, about 442 years ago). AIM OF THE STUDY: This work is purposed to investigate the pharmacological effects and mechanism of total flavonoids from Elaeagnus pungens leaves (FLA) on asthma in vivo and vitro. MATERIALS AND METHODS: Female BALB/c mice were sensitized by intraperitoneal injection of OVA with aluminum hydroxide and intranasal challenged with OVA. After treatment with FLA (10, 20 mg/kg p.o.), the behaviors of mice were observed by score evaluation. Enumeration of total cells and OVA-specific IgE assay in the blood were measured as well as enumeration of total cells and cytokines assay in the BALF. Furthermore, histopathological analysis was performed by HE staining. The in vitro relaxing action on muscle force of FLA (0.0316-10.0 mg/ml) was evaluated using isometric tension in tracheal rings, and VDLCC currents were recorded to explore the relaxation mechanism in the isolated tracheal rings and mouse ASM cells, respectively. In vitro anti-inflammatory actions were assessed with LPS-stimulated RAW 264.7 macrophages. The production of inflammatory mediators and MAPK signaling pathway was estimated using ELISA and Western blotting analysis, respectively. RESULTS: The high dose of flavones from E. pungens leaf (20 mg/kg) can significantly improve the symptom of asthma breakout and relieve the lung swelling. FLA treatment decreased eosinophils and leukocytes numbers in blood and BLAF with a dosedependent manner. Furthermore, the inhibiting effect of FLA on the level of Ig E and inflammatory-related cytokines including TNF-α, IL-5 showed dose-independent. FLA relaxed high K + -induced contraction in a dose-dependent manner. The maximal relaxation produced by FLA was 99.7% (IC 50 = 0.46 mg/ml). The whole-cell VDLCC currents were abolished by FLA (3.16 mg/ml) and FLA significantly decreased the maximal amplitude of VDLCCs. No cytotoxic effect of FLA was observed in RAW264.7 cells under the tested concentrations (1-300 µg/mL). The increased IL-6 and NO by the stimulation of LPS in RAW264.7 cells were significantly inhibited by FLA in the dosedependent manner. Treatment with LPS in the presence of FLA, LPS-induced phosphorylation of ERK1/2 and JNK was inhibited in the macrophages. CONCLUSION: FLA from Elaeagnus pungens leaf can alleviate the inflammation symptom via reducing the eosinophils and leukocytes numbers as well as the production of pro-inflammatory cytokines. This anti-inflammatory effect is related to the modulation of the MAPK signaling pathway. FLA can relax the precontracted TRs by blocking the VDLCCs, which interrupts extracellular Ca 2+ influx and inhibit the rise of [Ca 2+ ]i. It strongly suggests that these flavonoids components are the substances basis of Elaeagnus pungens leaves for allergic action, bronchospasm and inflammation in asthma.

Delayed-Onset NOG Gene-Related Syndromic Conductive Deafness: A Case Report.

We report a 6-year-old girl with progressive bilateral conductive hearing loss for 2 years. She passed the newborn hearing screening conducted with otoacoustic emissions testing and had a normal development of speech and language, which indicated that her deafness was delayed-onset. She also had congenital proximal interphalangeal joints. Proximal symphalangism was confirmed by genetic testing (NOG gene: c.406C > T, p.R136C). Bilateral stapes ankyloses were proved by surgery and her hearing was improved after stapedotomy by over 30 dB. Besides, this case should remind clinicians to carefully distinguish NOG gene-related deafness from congenital ossicular malformation and pediatric otosclerosis.

Injectable Hypoxia-Induced Conductive Hydrogel to Promote Diabetic Wound Healing.

Injectable hydrogels with the capability to cast a hypoxic microenvironment is of great potentialities to develop novel therapies for tissue regeneration. However, the relative research still remains at the conceptual phase. Herein, we chose diabetic wound as a representative injury model to explore the actual therapeutic results of tissue injury by injectable hypoxia-induced hydrogels. To enhance recovery and widen applicability, the hypoxia-induced system was incorporated with a conductive network by an original sequentially interpenetrating technique based on the combination of a fast "click chemistry" and a slow enzymatic mediated cross-linking. Hyperbranched poly(ß-amino ester)-tetraaniline (PBAE-TA) was cross-linked with thiolated hyaluronic acid (HA-SH) via a thiol-ene click reaction, contributing to the rapid formation of the first conductive network, where vanillin-grafted gelatin (Geln-Van) and laccase (Lac) with a slow cross-linking rate were employed in casting a hypoxic microenvironment. The as-prepared injectable hydrogels possessed both suitable conductivity and sustainable hypoxia-inducing capability to upregulate the hypoxia-inducible factor-1α and connexin 43 expressions of the encapsulated adipose-derived stem cells, which enhanced vascular regeneration and immunoregulation and further promoted the reconstruction of blood vessels, hair follicles, and dermal collagen matrix, eventually leading to the recovery of diabetic rat skin wounds and restoration of skin functions. This work provides a promising strategy to broaden the applicability of diverse hydrogels with a long time-consuming gelation process and to integrate different networks with various biological functions for the therapies of diabetic wounds and other complex clinical symptoms.