Search

Objective: To better understand the clinical phenotype of Ménière's disease (MD), we examined family history, thyroid disorder, migraine, and associated disorders in complaints of people living with MD. Method: We designed the study as a retrospective and examined data gathered from 912 participants with MD. Their data were originally collected by the Finnish Ménière Federation (FMF). The survey data included individual case histories for environmental factors, comorbidities, disease-specific complaints, impact-related questions, cognitive complaints, health-related quality of life (HRQoL), and sense of coherence (SOC). Results: We observed significant differences between those with and without sporadic occurrence, family history, thyroid disorder, and migraine-associated complaints. Family history explained 20% of variability in patient complaints. Patients with a family history of MD whose disease started at younger age experienced balance problems, more severe vertigo spells, more severe vestibular drop attacks (VDA), and less nausea, although they had good SOC. Thyroid disorder explained 14% of variability in patient complaints. MD patients with a thyroid disorder comorbidity suffered more often from constant dizziness, balance problems, greater impact of hearing problems, cognitive complaints, and poor HRQoL. Migraine explained 12% of variability in patients’ complaints and was associated with poor SOC and cognitive balance problems. MD patients with both thyroid disorder and migraine used antidepressants more often than other groups. Logistic regression analysis showed comorbidities of ischemic brain disorder (among 7.1%), kidney insufficiency (among 1.2%), and diabetes (among 7.3%) had statistically significant but restricted association with balance and gait problems, VDA, and reduced HRQoL. Conclusions: Family history of MD and thyroid disorder or migraine comorbidities in MD influence the complaint pattern and partially explain complex symptom profiles, including symptoms of cognitive problems. Confounders play a minimal role in complaint profile and impact of MD whereas comorbidities influence the complaint structure and partly explain the complex symptom profile in MD.

A novel bifunctional single-sheet electromagnetic surface is proposed in this paper. To alleviate the mutual coupling of the two functions, an equivalent series circuit for linear-to-linear polarization conversion and an equivalent parallel circuit for filtering property are cascaded. Thanks to the combination of different equivalent circuits and the displacement of the resonating parts of two functions, the proposed electromagnetic surface demonstrates filtering and polarization conversion properties in adjacent frequency bands. When the unit cell is periodically arranged to constitute a finite metasurface, more than 6 dB RCS reduction is obtained from 7.7 GHz to 22.8 GHz. The physical mechanism reveals that the low RCS property of the metasurface is in virtue of both scattering field cancellation and transmission. The simple structure, relatively small cell size and good performance of the metasurface suggest it may be a promising candidate as antenna radome. Moreover, the proposed integration method of different functions in one surface makes the design of multifunctional electromagnetic surface easier and faster.

Plant stress responses are important mechanisms that allow plants to adapt and survive in a dynamic and often challenging climatic conditions. In nature, plants encounter a wide range of stressors, including both biotic and abiotic factors, which can negatively impact their growth, development, and overall fitness. Plants therefore need to develop coping mechanisms to these stressors. Key players in these regulatory processes include transcription factors, kinases, and various signalling molecules. MicroRNAs (miRNAs) are a class of small, non-coding RNA molecules that play a pivotal role in plant stress tolerance. MiRNAs are increasingly recognized as key players in orchestrating how plants and other organisms adapt to a multitude of environmental stresses. However, little is still known on the function of miRNAs in plant responses to multiple stresses. This systematic review aimed to provide a comprehensive overview and analysis of the current state of knowledge regarding the role of microRNAs (miRNAs) in plant responses to multiple stresses under the changing environmental. The review paper has synthesized the existing research, highlighted key findings, and offered insights into the significance of miRNAs in enhancing plant resilience to various environmental stressors. It has concluded that the exploration of microRNAs (miRNAs) in the context of enhancing plant resilience to multiple stresses has yielded valuable insights into the intricacies of plant stress responses and the potential applications of these small regulatory molecules in agriculture for improved productivity under different stress conditions.

Benzo[a]pyrene (BaP) is a polycyclic aromatic hydrocarbon compound that is generated during combustion processes, and is present in various substances such as foods, tobacco smoke, and burning emissions. BaP is extensively acknowledged as a highly carcinogenic substance to induce multiple forms of cancer, such as lung cancer, skin cancer, and stomach cancer. Recently it is shown to adversely affect the reproductive system. Nevertheless, the potential toxicity of BaP on oocyte quality remains unclear. In this study, we established a BaP exposure model via mouse oral gavage and found that BaP exposure resulted in a notable decrease in the ovarian weight, number of GV oocytes in ovarian, and oocyte maturation competence. BaP exposure caused ribosomal dysfunction, characterized by a decrease in the expression of RPS3 and HPG in oocytes. BaP exposure also caused abnormal distribution of the endoplasmic reticulum (ER) and induced ER stress, as indicated by increased expression of GRP78. Besides, the Golgi apparatus exhibited an abnormal localization pattern, which was confirmed by the GM130 localization. Disruption of vesicle transport processes was observed by the abnormal expression and localization of Rab10. Additionally, an enhanced lysosome and LC3 fluorescence intensity indicated the occurrence of protein degradation in oocytes. In summary, our results suggested that BaP exposure disrupted the distribution and functioning of organelles, consequently affecting the developmental competence of mouse oocytes.

Background: Emerging evidence has shown the potential involvement of phthalates (PAEs) exposure in the development of dementia with Lewy bodies (DLB). Metabolomics can reflect endogenous metabolites variation in the progress of disease after chemicals exposure. However, little is known about the association between PAEs, gut microbiota and metabolome in DLB. Objective: We aim to explore the intricate relationship among urinary PAEs metabolites (mPAEs), dysbiosis of gut bacteria, and metabolite profiles in DLB. Methods: A total of 43 DLB patients and 45 normal subjects were included in this study. Liquid chromatography was used to analyze the levels of mPAEs in the urine of the two populations. High-throughput sequencing and liquid chromatography-mass spectrometry were used to analyze gut microbiota and the profile of gut metabolome, respectively. The fecal microbiota transplantation (FMT) experiment was performed to verify the potential role of mPAEs on gut dysbiosis contribute to aggravating cognitive dysfunction in α-synuclein tg DLB/PD mice. Results: The DLB patients had higher DEHP metabolites (MEOHP, MEHHP and MEHP), MMP and MnBP, lower MBP and MBzP than the control group and different microbiota. A significantly higher abundance of Ruminococcus gnavus and lower Prevotella copri, Prevotella stercorea and Bifidobacterium were observed in DLB. Higher 3 DEHP metabolites, MMP, MnBP and lower MBP and MBzP were significantly negatively associated with Prevotella copri, Prevotella stercorea and Bifidobacterium. Additionally, using metabolomics, we found that altered bile acids, short-chain fatty acids and amino acids metabolism are linked to these mPAEs. We further found that FMT of fecal microbiota from highest DEHP metabolites donors significantly impaired cognitive function in the germ-free DLB/PD mice. Conclusion: Our study suggested that PAEs exposure may alter the microbiota-gut-brain axis and providing novel insights into the interactions among environmental perturbations and microbiome-host in pathogenesis of DLB.

Wasp venom injections from wasp stings can damage several organs, most commonly the kidneys. Despite literature evidence, wasp sting-induced acute kidney injury (AKI) is rare and involves complex pathophysiological processes. While acute tubular necrosis (ATN) is the most prevalent histological result of wasp sting-induced AKI, uncommon combinations of chronic renal lesions have been described, alerting us to the patient's underlying illness. We report a 55-year-old hypertensive patient with unknown renal function who got AKI following multiple wasp stings. His renal function had not improved after continuous hemodialysis and plasma exchange; therefore, a kidney biopsy was performed. The pathology revealed that in addition to ATN, his kidney's distinguishing feature was a mix of chronic interstitial renal disease and chronic glomerulosclerosis. We think that his current renal pathological results were caused by hypertension in addition to wasp venom.

The aim of the present work was to show the sustainability of fibrin sealant in releasing dexamethasone and adjust the protocol for clinical application of the novel method in the treatment of Meniere's disease (MD) and sudden sensorineural hearing loss (SSHL). Gelation occurred shortly after mixing dexamethasone-containing fibrinogen with thrombin. Dexamethasone was constantly released for at least 16 d at a stable level after 7 d in protocol 1 (low-dose), while it was robustly released within 4 d and slowed afterward until 10 d in protocol 2 (high-dose). There were significant differences among the time points in Protocol 2 (p

Understanding the evolution of Fe–containing intermetallic compounds (Fe–IMCs) in Al–Si–Mg alloys is essential for improving the mechanical performances. This paper aims to systemically evaluate the influences of both the single addition of Mn or Mo, and the combined additions of Mn and Mo on the evolution of Fe–IMCs and mechanical performances of Al–Si–Mg cast alloys. Results reveal that the as–cast microstructures of these alloys are primarily composed of Al3M (M means Ti, Zr, or TiZr), α–Al, eutectic Si, Mg2Si, Q–Al5Cu2Mg8Si6, and some irregular Fe–IMCs. It is clear that the morphology and size of Fe–IMCs are significantly affected by the single or combined addition of Mn and Mo. Moderate Mn content facilitates the formation of Chinese script–like α–Al(FeMn)Si phases that are favorable to the mechanical properties. The combined addition of Mn and Mo refines the α–Al(FeMn)Si phases and promotes the formation of Chinese script–like α–Al(FeMnMo)Si phases. Furthermore, as the Mo level increases, β–AlFeSi, π–Al8FeMg3Si6, and α–Al(FeMn)Si disappear successively, which also helps improve the mechanical performances. In this article, maximum hardness, ultimate tensile strength, yield strength and elongation of 87.04 HV, 275.34 MPa, 139.70 MPa, 7.88% are obtained when Fe, Mn and Mo are added at 0.20wt.%, 0.02wt.%, and 0.27wt.%, respectively. The purpose of this work is to provide an efficient method to minimize or even eliminate the adverse effects of Fe–IMCs and thus improve the mechanical performances of Al–Si–Mg cast alloys.

In this paper, we investigated the influence of calcium (Ca) on the electrochemical and corrosion properties of Al–Mg–Sn–Ga alloys in neutral and alkaline electrolytes, respectively. Results indicate that Ca element promotes the uniform dissolution of the anode and improve its discharge efficiency. The peak energy densities of the Al-0.5Mg-0.1Sn-0.05Ga-0.1Ca alloy in neutral and alkaline solutions are 3361 and 3581 Wh kg−1, respectively. Compared with calcium-free alloy, the increase is 5.3% and 26.5%, respectively. Additionally, the severe intergranular corrosion behavior of Al-0.5Mg-0.1Sn-0.05Ga alloy can be significantly inhibited by adding Ca element under alkaline conditions. Therefore, the addition of Ca element further optimizes the discharge performance of Al–Mg–Sn–Ga-based anodes and enhances its environmental adaptability.

Aim or Purpose: This study aimed to investigate the effects of silicone samples with different fluoride concentrations on the growth and biofilm formation of Streptococcus mutans (S. mutans). Materials and Methods: The experimental samples, with a diameter of 10 mm and a thickness of 2.5 mm, were prepared using mixtures of silicone and sodium fluoride with sodium fluoride concentrations of 0.5%, 1%, 1.5%, 2%, and 2.5%, respectively. The control group did not contain sodium fluoride. Each group had 6 pieces. All samples were immersed in 2 ml of deionized water with pH=7.0, and then the extract was added to S. mutans culture medium. Optical density values at 600 nm (OD600) at 5, 8, and 12 hours and 575 nm (OD575) at 4, 8, and 16 hours were measured to evaluate the growth and biofilm formation of S. mutans, respectively. Results: At all time-points, the OD600 value in all experimental groups were significantly lower compared to the control group (P

High-throughput double cone specimens were used to study the influence of Mo on the microstructure evolution of Al–Cu–Mg–Ag alloy under a wide range of strain along the axis. The microstructure evolution was characterized by scanning electron microscopy (SEM), transmission electron microscopy (TEM) and electron backscatter diffraction (EBSD). Results demonstrated that Al6(Fe, Mn, Mo) dispersoids formed in the Al–Cu–Mg–Ag alloy with the addition of 0.2% Mo during double-step homogenization. Meanwhile, rod-like Al20Cu2Mn3 and disk-like Al6(Fe, Mn, Mo) dispersoids segregated to opposite distribution in interdendritic regions and dendrite cores, respectively, which obviously eliminated the dispersoid-free zones and intragranular sparse regions. With different effective strains along the axis, Al6(Fe, Mn, Mo) dispersoids obviously reduced the transformation from Low-angle grain boundaries to High-angle grain boundaries. The nucleation and growth of dynamic recrystallization grains were effectively inhibited by the coupling of Al20Cu2Mn3 and Al6(Fe, Mn, Mo) dispersoids due to the obstruction of dislocation movement.

The etiology and underlying mechanism of Meniere's disease (MD) development are still unknown, although inflammation and autoimmunity have been implicated as underlying mechanisms. The human endolymphatic sac (ES) has been reported to have innate and adaptive immune capacity in local immune reactions. In vivo demonstration of inflammation of the ES in patients with MD is missing in the literature. We report the case of a 47-year-old female patient diagnosed with unilateral MD with genetic variants and cytokine markers indicating inflammation and vascular congestion of the ES. Endolymphatic hydrops in the right cochlea (grade 2) and vestibulum (grade 3) were detected using MRI. She carried heterozygous variants in MEFV (c.442G > C), IRF8 (c.1157G > T), ADA (c.445C > T), PEPD (c.151G > A), NBAS (c.4049T > C), CSF2RB (c.2222C > T), HPS6 (c.277G > T), IL2RB (c.1109C > T), IL12RB1 (c.1384G > T), IL17RC (c.260_271del GCAAGAGC TGGG), LIG1 (c.746G > A), RAG1 (c.650C > A), and SLX4 (c.1258G > C, c.5072A > G). In the serum, the levels of granulocyte colony-stimulating factor (G-CSF), macrophage inflammatory protein 1α, and IL7 were significantly elevated, and the level of IL2Rα was reduced. Intratympanic administration of dexamethasone temporarily alleviated her hearing loss. Her vertigo was significantly relieved but remained slight after ES administration of corticosteroids.

Background: Intratympanic administration of gadolinium chelate allows for a better visualization of endolymphatic hydrops (EH) using MRI than intravenous injection and was recently further improved to facilitate high-quality imaging of EH in 7 min. The aim of the present study was to simplify the intratympanic administration protocol by mixing gadolinium chelate with therapeutic dexamethasone and to evaluate the effects of this mixture on the visualization of EH in MRI. Materials and methods: In an in vitro study, the potential impact of gadolinium-diethylenetriamine pentaacetic acid (Gd-DTPA) on the stability of dexamethasone was evaluated by analyzing dynamic changes in dexamethasone with high-performance liquid chromatography (HPLC) after mixing with Gd-DTPA. Ten patients with definite Meniere's disease (MD) were recruited to study the potential interference of dexamethasone on MRI visualization of EH, and 49 patients with MD were recruited to evaluate the effect of intratympanic injection of Gd-DTPA mixed with dexamethasone on MRI of EH using a 3T MR machine and a novel heavily T2-weighted 3-dimensional fluid-attenuated inversion recovery reconstructed using a magnitude plus zero-filled interpolation (hT2FLAIR-MZFI) sequence. Results: The retention times and peak area of dexamethasone in HPLC were not modified by the addition of Gd-DTPA. EH grading in the cochlea and vestibule was not influenced in any ear by intratympanic injection of dexamethasone. Excellent inner ear images were obtained from all patients, and EHs with various grades were displayed. There were significant correlations between diagnosis and cochlear EH (p

The bile acid-responsive G-protein-coupled receptor TGR5 is expressed in monocytes and macrophages, and plays a critical role in regulating inflammatory response. Our previous work has shown its role in promoting the progression of non-small cell lung cancer (NSCLC), yet the mechanism remains unclear. Here, using Tgr5-knockout mice, we show that TGR5 is required for M2 polarization of tumor-associated macrophages (TAMs) and suppresses antitumor immunity in NSCLC via involving TAMs-mediated CD8+ T cell suppression. Mechanistically, we demonstrate that TGR5 promotes TAMs into protumorigenic M2-like phenotypes via activating cAMP-STAT3/STAT6 signaling. Induction of cAMP production restores M2-like phenotypes in TGR5-deficient macrophages. In NSCLC tissues from human patients, the expression of TGR5 is associated with the infiltration of TAMs. The co-expression of TGR5 and high TAMs infiltration are associated with the prognosis and overall survival of NSCLC patients. Together, this study provides molecular mechanisms for the protumor function of TGR5 in NSCLC, highlighting its potential as a target for TAMs-centric immunotherapy in NSCLC.

Bone morphogenetic protein 2 (BMP2) effectively induced mesenchymal stem cells (MSCs) osteogenic differentiation hold great potential for bone tissue engineering. However, a global mechanistic view of BMP2-induced osteogenic differentiation of MSCs remains to be fully elucidated. Here, human umbilical cord-derived MSCs (UC-MSCs) were induced with BMP2, three days and five days later, total RNA were extracted and subjected to RNA-sequencing (RNA-Seq) followed with bioinformatic analysis. Osteogenic differentiation abilities were evaluated with Alkaline phosphatase (ALP) staining and osteogenic differentiation marker expression at both mRNA and protein levels. We identified that adenoviral vectors effectively transduced in UC-MSCs and expressed BMP2 in high efficiency. Both on day 3 and day 5, differentially expressed genes (DEGs) were highly enriched in PI3K-Akt signaling pathway. As for the common DEGs among total BMP2 group vs control group, BMP2 (day 3) versus control (day 3) and BMP2 (day 5) versus control (day 5), there were 105 DGEs and highly enriched in PI3K-Akt signaling pathway. Finally, we found that PI3K-Akt signaling inhibitor dramatically inhibited BMP2-iduced osteogenic differentiation of UC-MSCs. We firstly identified that PI3K-Akt signaling pathway plays a pivotal role in BMP2-induced osteogenic differentiation of MSCs, which may apply a new perspective for BMP2 based bone tissue engineering.

Fine, uniform and equiaxed grain structure is one of the most striking features of high-quality Al targets. In general, the imposition of magnetic stirring on the liquid Al during the solidification process is an effective method to promote grain refinement. In the present study, a self–made rotating system of permanent magnets is reported to stir liquid Al during solidification. The temperature change and grain structure of pure Al subjected to a permanent magnet stirring (PMS) are analyzed. The experimental results reveal that the forced convection driven by PMS can lead to a more homogeneous temperature field in the melt. Meanwhile, PMS can greatly refine the grain structure of pure Al and eliminate cavities, but it may induce more gas holes in the ingots. Moreover, a fresh intermittent permanent magnet stirring (I-PMS) approach has been proposed and successfully applied to realize grain refinement of pure Al. Some casting problems can be perfectly solved using suitable processing parameters. Through some valuable experimental works, systematic researches on grain refinement of pure Al under PMS are performed to better understand its refining mechanisms. Therefore, this article further enhances our understanding of related fields and fills the research blank.

Crowded living conditions and increasingly severe noise problems necessitate the development of a low-thickness sound absorption material with remarkable sound absorption performance. Here, simple materials such as sodium alginate aerogel (SA) and polyurethane (PU) foam are used to combine with special 3D spacer fabric to design an innovative highly-efficient acoustic absorber composite (B&M−L composite). It integrates a “backing cavity-like” (BC-L) structure and “filled-microperforated plate-like” (MPP-L) structure. Thereby, it realizes porous and resonant sound absorption at a small thickness. The superior structure design combining strong resonance with porous structure endows the new type of composite with a remarkable absorption performance. Specifically, the peak absorption coefficient is 0.98, the average absorption coefficient at a thickness of 10 mm is 0.71, and the noise reduction coefficient per unit thickness of B&M−L composites exceeds those of most of the reported sound absorption materials. Based on its remarkable acoustic performance, good mechanical properties, heat insulation performance, and thermal stability, the B&M−L composite provides a convenient and inexpensive method to improve the overall performance of porous sound absorption materials, and displays significant potential for use in the fields of construction, transportation, and large mechanical equipment.

Retinoblastoma (RB) is the most common intraocular tumor among children. Leucine-rich pentatricopeptide repeat (PPR)-motif-containing protein (LRPPRC), a suppressor gene of autophagy, has been proven to play a regulatory role in tumor progression. However, little is known about functional roles and mechanisms of LRPPRC in RB progression. First, we performed a detailed analysis for RB and normal control. The expression of LRPPRC in the RB tissues was significantly higher than that in normal tissues. Moreover, LRPPRC suppression could repress tumor cell migration, invasion, glycolysis, and reactive oxygen species (ROS)/hypoxia-inducible factor-1α (HIF1-α) pathway activation by mediating autophagy. Furthermore, overexpression of HIF1-α partially reversed the above changes induced by LRPPRC knockdown. The regulation of LRPPRC on tumor metastasis and glycolysis was also validated by a xenograft tumor assay. In summary, LRPPRC could regulate metastasis and glycolysis of RB by mediating autophagy suppression and further activating the ROS/HIF1-α pathway, and LRPPRC could be a promising prognostic biomarker for RB.

A novel technology, termed intermittent permanent magnet stirring (I-PMS), is proposed to improve the quality of aluminum alloy ingots. In this paper, the melt flow and solidification structure of Al–4.5Cu alloy were investigated through I-PMS. The grain refinement mechanism of I-PMS is also discussed. The results indicate that the melt flow patterns and distributions can be greatly affected by I-PMS. When I-PMS is applied during solidification, the melt undergoes a series of complicated periodic three-dimensional flows. Compared without I-PMS, the solidification structure of Al–4.5Cu alloy with I-PMS was remarkably refined, and the optimum solidification structure could be realized utilizing appropriate processing parameters.

Excitonic effects induced via Coulomb interactions and an appropriate interfacial contact of electrode modified materials play crucial roles in transfer/separation of interfacial charge carriers during the electrochemical sensing process. Herein, an oxygen vacancy (Vo) mediated step-scheme (S-scheme) heterojunction of WO2.9/g-C3N4 was fabricated by electrostatic self-assembly of high-purity commercial WO2.9 nanoparticles and ultrathin g-C3N4 nanosheets, where g-C3N4 nanosheets were prepared via polymerization followed ultrasonic-assisted exfoliation. The fabricated Vo-S-scheme WO2.9/g-C3N4 heterojunction-based sensor exhibited superior electrochemical performance for detecting environmental hormone 4-nitrophenol (4-NP). Due to the introduction of Vo, an increased free-charge-carriers, a built-in electric field and a high adsorption energy are generated at the heterojunction interface, which all can significantly improve the sensitivity of this proposed sensor. It also showed good reproducibility, high accuracy, good recovery, and strong feasibility for 4-NP detection in real samples. This work not only conduces significantly to understanding the defective structures and S-scheme heterojunctions, but also provides new insights into designing and constructing novel efficient electrochemical sensors.

Objectives: To evaluate the pathological changes in the blood-perilymph, blood-endolymph, and blood-nerve barriers of a patient with idiopathic sudden sensorineural hearing loss (SSNHL). Methods: Potential ossification or fibrosis in the inner ear was evaluated using temporal bone CT and MRI acquired using the 3-dimensional T2-weighted sampling perfection with application-optimized contrasts using a flip angle evolution sequence. Pathological changes in the barriers were analyzed by MRI obtained 4 h after a single-dose intravenous injection of gadolinium chelate using a medium inversion-time inversion recovery imaging with magnitude reconstruction sequence. Results: The perilymph was absent, while significant enhancements of the vestibulocochlear nerve and the endolymphatic compartments were detected. Conclusion: Significant injuries in the blood-endolymph and blood-vestibulocochlear nerve barriers and disabled perilymph production may contribute to the development of SSNHL with poor response to treatments.

Objective: To investigate immune-related genetic background in intractable Meniere’s disease (MD) and the immediate results of a novel therapy by delivering steroids to the surface of the intact endolymphatic sac (ES) and incus in a sustainable manner. Case report and methods: Candidate genes involved in immune regulation were sequenced using a next-generation sequencing method in a patient with intractable MD. Mutations were confirmed using the Sanger sequencing method. The ES was exposed, and gelatin sponge particles were immersed in high-dose methylprednisolone solution and placed onto the surface of ES. “L”-shaped gelatin sponge strips were immersed in dexamethasone solution and served as a guiding device for the steroids by touching the incus and gelatin sponge particles on the surface of the ES. Gelatin sponge particles immersed in dexamethasone solution were placed around the gelatin sponge strips and sealed using fibrin glue. Results: Autoinflammation in the refractory MD case was indicated by genotype, including novel heterozygous mutations of PRF1, UNC13D, SLC29A3, ITCH, and JAK3, as well as phenotype. The vertigo was fully relieved immediately after operation. Tinnitus and aural fullness were resolved 3 weeks after operation, whereas hearing improved in 2 mon postoperation. No recurrence was noted during the 5-monfollow-up, and the final MRI supported the novel therapeutic hypothesis. Conclusion: Autoinflammation was involved in a refractory MD. This novel therapy, which involves the delivery of steroids to the surface of the intact ES and incus, is effective in relieving vertigo and tinnitus and improves hearing function of refractory MD. Keywords: Vestibular function, Tinnitus, Genetic disorders, Hearing

CRISPR/Cpf1 has emerged recently as an effective tool for genome editing in many organisms, but its use in pigs to generate precise genetic modifications has seldom been described. Myostatin (MSTN) is a well-characterized negative regulator of muscle development, and natural mutations in this gene cause a double-muscled phenotype in many species. However, to the best of our knowledge, no naturally occurring mutation in MSTN has been found in pigs. In addition, no living pig models with sophisticated modifications orthologous to natural mutations in MSTN have yet been reported. In this study, we exploited the CRISPR/Cpf1 system to introduce a predefined modification orthologous to the natural MSTN mutation found in Belgian Blue cattle (thus known as the Belgian Blue mutation). Our research demonstrated that the cutting efficiency of CRISPR/Cpf1 was 12.3% in mixed porcine fetal fibroblasts in drug free medium, and 41.7% in clonal colonies obtained using G418 selection. Then, the Cpf1-sgRNA vector, ssODN template, and a self-excision cassette were co-transfected into porcine fetal fibroblasts. After G418 selection, 8 clonal colonies were examined and 5 with genetic modification were found. Of these 5, 2 harbored the precise 11-bp deletion. Using 1 heterozygous clonal colony, 2 cloned Duroc piglets were successfully generated, which was heterozygous for the Belgian Blue mutation. In summary, our results demonstrate that CRISPR/Cpf1 system can be used efficiently to generate double-stranded breaks, and also to mediate homologous recombination to introduce precise genomic modifications in pigs.

Background: Many flaviviruses are significant human pathogens that cause global public health threats. Developing research tools for studying and diagnosing these pathogens is a top priority. Reporter flaviviruses are useful tools for studying viral pathogenesis, diagnosing disease, and screening antiviral compounds. However, the stability of reporter flaviviruses has been challenged by viral RNA recombination, leading to deletion of the engineered reporter gene during viral replication. The instability of reporter viruses has limited their application to research and countermeasure development. Thus, new approaches to overcome the instability of reporter flaviviruses are critically needed to advance the flavivirus field. Methods: To create a stable flavivirus bearing a reporter gene, we engineered mutations in the viral capsid gene that are rendered virus-lethal upon recombination. Thus, only non-recombined reporter virus propagates. We tested this strategy using Zika virus (ZIKV) bearing a nano-luciferase (NanoLuc) gene and passaged both virus with capsid mutations and virus without mutations. Findings: The recombination-dependent lethal mutations succeeded in stabilizing the NanoLuc ZIKV through ten passages, while WT reporter virus showed instability as early as five passages. The stability of NanoLuc ZIKV was supported by RT-PCR, sequencing, focus forming assay, and luciferase assay. The success of this method was reconfirmed by also establishing a stable NanoLuc Yellow Fever 17D virus, indicating that the recombination-dependent lethal approach can be applied to other flaviviruses. To demonstrate the utility of the stable reporter viruses, we showed that NanoLuc ZIKV and YFV17D could be used to measure neutralizing antibody titers with a turnaround time as short as four hours. Importantly, the neutralizing antibody titers derived from the reporter virus assay were equivalent to those derived from the conventional plaque assay, indicating the new assay maintains the gold standard of serology testing. Furthermore, using a known inhibitor, we showed that the reporter viruses could be reliably used for antiviral evaluation. Interpretation: The study has developed a recombination-dependent lethal approach to produce stable reporter flaviviruses that may be used for rapid serodiagnosis, trans-gene delivery, vaccine evaluation, and antiviral discovery. Funding: National Institute of Health, Robert J. Kleberg, Jr. and Helen C. Kleberg Foundation; John S. Dunn Foundation; Amon G. Carter Foundation; Gillson Longenbaugh Foundation; Summerfield G. Roberts Foundation.

Background: Vaccines are the most effective means to fight and eradicate infectious diseases. Live-attenuated vaccines (LAV) usually have the advantages of single dose, rapid onset of immunity, and durable protection. DNA vaccines have the advantages of chemical stability, ease of production, and no cold chain requirement. The ability to combine the strengths of LAV and DNA vaccines may transform future vaccine development by eliminating cold chain and cell culture with the potential for adventitious agents. Methods: A DNA-launched LAV was developed for ZIKV virus (ZIKV), a pathogen that recently caused a global public health emergency. The cDNA copy of a ZIKV LAV genome was engineered into a DNA plasmid. The DNA-LAV plasmid was delivered into mice using a clinically proven device TriGrid™ to launch the replication of LAV. Findings: A single-dose immunization as low as 0.5 μg of DNA-LAV plasmid conferred 100% seroconversion in A129 mice. All seroconverted mice developed sterilizing immunity, as indicated by no detectable infectious viruses and no increase of neutralizing antibody titers after ZIKV challenge. The immunization also elicited robust T cell responses. In pregnant mice, the DNA-LAV vaccination fully protected against ZIKV-induced disease and maternal-to-fetal transmission. High levels of neutralizing activities were detected in fetal serum, indicating maternal-to-fetal humoral transfer. In male mice, a single-dose vaccination completely prevented testis infection, injury, and oligospermia. Interpretation: The remarkable simplicity and potency of ZIKV DNA-LAV warrant further development of this vaccine candidate. The DNA-LAV approach may serve as a universal vaccine platform for other plus-sense RNA viruses. Fund: National Institute of Health, Kleberg Foundation, Centers for Disease Control and Prevention, University of Texas Medical Branch. Keywords: Zika virus, DNA vaccine, live-attenuated vaccine, flavivirus

Vestibular schwannoma (VS) is a slow-growing benign neoplasm. There has been an evolution in the management of VS from active treatments (microsurgery and stereotactic radiotherapy) to conservative management (wait and scan). Regular MRI scanning is necessary to monitor tumor progression. Conservative management causes significantly less complications and offers a higher quality of life compared with active treatments. The mean growth rate of VS varies from 0.4 to 2.9 mm/year, and spontaneous shrinkage is observed in 3.8 percent of tumors during observation. If significant growth occurs, active treatment is considered. Significant growth is defined as an increase of at least 3 mm in the largest extrameatal diameter in any plane between the first and last available scans. The vestibulocochlear nerve is surrounded by cerebrospinal fluid, which provides natural contrast for MRI; thus, gadolinium may not be needed to detect VS. Specific sequences have high sensitivity, specificity, and accuracy for detection of progression. Hypointense signal in the ipsilateral inner ear fluid might be a useful sign to distinguish VS from meningioma. In this paper, we summarize the current status of research on conservative management and non-contrast MRI for the detection of VS.

Summary: Zika virus (ZIKV) infection causes devastating congenital abnormities and Guillain-Barré syndrome. The ZIKV envelope (E) protein is responsible for viral entry and represents a major determinant for viral pathogenesis. Like other flaviviruses, the ZIKV E protein is glycosylated at amino acid N154. To study the function of E glycosylation, we generated a recombinant N154Q ZIKV that lacks the E glycosylation and analyzed the mutant virus in mammalian and mosquito hosts. In mouse models, the mutant was attenuated, as evidenced by lower viremia, decreased weight loss, and no mortality; however, knockout of E glycosylation did not significantly affect neurovirulence. Mice immunized with the mutant virus developed a robust neutralizing antibody response and were completely protected from wild-type ZIKV challenge. In mosquitoes, the mutant virus exhibited diminished oral infectivity for the Aedes aegypti vector. Collectively, the results demonstrate that E glycosylation is critical for ZIKV infection of mammalian and mosquito hosts. : Zika virus (ZIKV) causes devastating congenital abnormities and Guillain-Barré syndrome. Fontes-Garfias et al. showed that the glycosylation of ZIKV envelope protein plays an important role in infecting mosquito vectors and pathogenesis in mouse. Keywords: Zika virus, glycosylation, flavivirus entry, mosquito transmission, vaccine

Objective: To test the feasibility of measuring fine temporal bone structures using a newly established cone-beam computed tomography (CBCT) system. Materials and methods: Six formalin-fixed human cadaver temporal bones were imaged using a high-resolution CBCT system that has 900 frames and copper + aluminum filtration. Fine temporal bone structures, including those of the facial nerve canal and vestibular structures, were identified and measured. Results: The fine structures of the middle ear, including the tympanic membrane, tendon of the tensor tympani, cochleariform process of the semicanal of the tensor tympani, pyramidal eminence, footplate of the stapes, full path of the facial nerve within the temporal bone, supralabyrinthine space, semicircular canals, pathway of the subarcuate canal, and full path of the vestibular aqueduct, were clearly demonstrated. The vestibular aqueduct has a midpoint width of 0.4 ± 0.0 mm and opercular width of 0.5 ± 0.1 mm (mean ± SD). The length of the internal acoustic meatus was 10.6 ± 1.2 mm (mean ± SD), and the diameter of the internal acoustic meatus was 3.7 ± 0.3 mm (mean ± SD). Conclusion: This novel high-resolution CBCT system has potentially broad applications in the diagnosis of inner ear disease and in monitoring associated pathological changes, surgical planning, navigation for the ear surgery, and temporal bone training.

A purified inactivated vaccine (PIV) using the Zika virus (ZIKV) Puerto Rico strain PRVABC59 showed efficacy in monkeys, and is currently in a phase I clinical trial. High-yield manufacture of this PIV is essential for its development and vaccine access. Here we report an infectious cDNA clone-launched platform to maximize its yield. A single NS1 protein substitution (K265E) was identified to increase ZIKV replication on Vero cells (a cell line approved for vaccine production) for both Cambodian FSS13025 and Puerto Rico PRVABC59 strains. The NS1 mutation did not affect viral RNA synthesis, but significantly increased virion assembly through an increased interaction between NS1 and NS2A (a known regulator of flavivirus assembly). The NS1 mutant virus retained wild-type virulence in the A129 mouse model, but decreased its competence to infect Aedes aegypti mosquitoes. To further increase virus yield, we constructed an infectious cDNA clone of the clinical trial PIV strain PRVABC59 containing three viral replication-enhancing mutations (NS1 K265E, prM H83R, and NS3 S356F). The mutant cDNA clone produced >25-fold more ZIKV than the wild-type parent on Vero cells. This cDNA clone-launched manufacture platform has the advantages of higher virus yield, shortened manufacture time, and minimized chance of contamination.

Rapid and accurate diagnosis of infectious agents is essential for patient care, disease control, and countermeasure development. The present serologic diagnosis of Zika virus (ZIKV) infection relies mainly on IgM-capture ELISA which is confounded with the flaw of cross-reactivity among different flaviviruses. In this communication, we report a multiplex microsphere immunoassay (MIA) that captures the diagnostic power of viral envelope protein (that elicits robust, yet cross-reactive antibodies to other flaviviruses) and the differential power of viral nonstructural proteins NS1 and NS5 (that induce more virus-type specific antibodies). Using 153 patient specimens with known ZIKV and/or dengue virus (DENV; a closely related flavivirus) infections, we showed that (i) ZIKV envelope-based MIA is equivalent or more sensitive than IgM-capture ELISA in diagnosing ZIKV infection, (ii) antibody responses to NS1 and NS5 proteins are more ZIKV-specific than antibody response to envelope protein, (iii) inclusion of NS1 and NS5 in the MIA improves the diagnostic accuracy when compared with the MIA that uses envelope protein alone. The multiplex MIA achieves a rapid diagnosis (turnaround time

There are several challenges to inner ear drug delivery and imaging due to the existence of tight biological barriers to the target structure and the dense bone surrounding it. Advances in imaging and nanomedicine may provide knowledge for overcoming the existing limitations to both the diagnosis and treatment of inner ear diseases. Novel techniques have improved the efficacy of drug delivery and targeting to the inner ear, as well as the quality and accuracy of imaging this structure. In this review, we will describe the pathways and biological barriers of the inner ear regarding drug delivery, the beneficial applications and limitations of the imaging techniques available for inner ear research, the behavior of engineered nanomaterials in inner ear applications, and future perspectives for nanomedicine-based inner ear imaging.

The current epidemic of Zika virus (ZIKV) has underscored the urgency to establish experimental systems for studying viral replication and pathogenesis, and countermeasure development. Here we report two ZIKV replicon systems: a luciferase replicon that can differentiate between viral translation and RNA synthesis; and a stable luciferase replicon carrying cell line that can be used to screen and characterize inhibitors of viral replication. The transient replicon was used to evaluate the effect of an NS5 polymerase mutation on viral RNA synthesis and to analyze a known ZIKV inhibitor. The replicon cell line was developed into a 96-well format for antiviral testing. Compare with virus infection-based assay, the replicon cell line allows antiviral screening without using infectious virus. Collectively, the replicon systems have provided critical tools for both basic and translational research.

The underground water-sealed storage technique is critically important and generally accepted for the national energy strategy in China. Although several small underground water-sealed oil storage caverns have been built in China since the 1970s, there is still a lack of experience for large-volume underground storage in complicated geological conditions. The current design concept of water curtain system and the technical instruction for system operation have limitations in maintaining the stability of surrounding rock mass during the construction of the main storage caverns, as well as the long-term stability. Although several large-scale underground oil storage projects are under construction at present in China, the design concepts and construction methods, especially for the water curtain system, are mainly based on the ideal porosity medium flow theory and the experiences gained from the similar projects overseas. The storage projects currently constructed in China have the specific features such as huge scale, large depth, multiple-level arrangement, high seepage pressure, complicated geological conditions, and high in situ stresses, which are the challenging issues for the stability of the storage caverns. Based on years' experiences obtained from the first large-scale (millions of cubic meters) underground water-sealed oil storage project in China, some design and operation problems related to water curtain system during project construction are discussed. The drawbacks and merits of the water curtain system are also presented. As an example, the conventional concept of “filling joints with water” is widely used in many cases, as a basic concept for the design of the water curtain system, but it is immature. In this paper, the advantages and disadvantages of the conventional concept are pointed out, with respect to the long-term stability as well as the safety of construction of storage caverns. Finally, new concepts and principles for design and construction of the underground water-sealed oil storage caverns are proposed.

Objective: To investigate immune-related genetic background in bilateral sudden sensorineural hearing loss (SSNHL). Case report and methods: The case is a 45-year-old man presenting with a 7-year history of bilateral profound SSNHL. Blood biochemical testing demonstrated increased levels of total cholesterol (5.88 mmol/L). Tests for hepatitis B showed a positive antibody against the hepatitis B core antigen. Complement C3 was below the normal value, and complement C4 and IgG were in the lower range of normal values. CT images showed a normal inner ear and vestibular aqueduct but round window membranous ossification on both sides. A total number of 232 immune-associated genes were sequenced using the next generation sequencing technique. Results: Mutations were detected in 5 genes, including the phosphoinositide 3-kinase catalytic subunit delta (PIK3CD), caspase recruitment domain-containing protein 9 (CARD9), complement factor H-related (CFHR2), immunoglobulin lambda-like polypeptide 1 Protein (IGLL1), and transmembrane channel-like gene family 8 (TMC8). In the PIK3CD gene, a C896T substitute in exon 7 was detected. This mutation causes primary immunodeficiency and is an autosomal dominant disease. Conclusion: The PIK3CD C896T mutation responsible for primary immunodeficiency may contribute to the onset of bilateral SSNHL with subsequent rapid progression.

More detailed knowledge of root and canal morphology is important in order to improve the success in the endodontics of primary teeth. The purpose of this study was to evaluate the root and canal morphology of primary mandibular second molars (PMSMs) in a Chinese population using cone-beam computed tomography (CBCT). Methods: CBCT images, which had been obtained previously in the West China Hospital of Stomatology at Sichuan University, Chengdu, China between May 2009 and December 2011, were screened retrospectively. Finally, 283 individuals—207 male and 76 female with a mean age of 7.2 years (range 3–10 years)—and 487 PMSMs with clear images of root and canal morphology were enrolled. The number of roots and morphology of canals were recorded. The patient's gender, and the symmetry and frequency of three roots in PMSMs were analyzed. Results: The majority of PMSMs had two (72.28%) or three roots (27.52%). The symmetrical incidence of three-root PMSMs in this Chinese population was 50.65%. There is no difference between genders in the prevalence of an extra root and the incidence of symmetry (p > 0.05). Of the individuals enrolled, 25.26% of PMSMs had three canals and 73.31% had four canals. The root canal systems of the PMSMs in the present study were categorized into seven variants. Conclusion: This study indicated that three-rooted PMSMs occur frequently in the Chinese population. There was no difference between the two genders on incidence and symmetry. The majority of PMSMs have three to four canals and the diversity of the root canal variants should be considered when performing clinical procedures. CBCT is a good option for studying the root and canal morphology of primary teeth.