Socioeconomic Status Impacts the Prognosis of Chronic Rhinosinusitis Treated by Endoscopic Sinus Surgery:An Observational Cohort Study in Northeast China

Objective To explore the association between socioeconomic status(SES)and postoperative outcomes in patients with chronic sinusitis(CRS)after functional endoscopic sinus surgery(ESS).Methods We conducted an observational cohort study of 1,047 patients with CRS undergoing ESS.Discharged patients were followed up to 72 weeks for all-cause recurrence events.Baseline SES was established based on occupation,education level,and family income of the patients 1 year before the operation.Kaplan–Meier method was used to calculate the recovery rate after ESS,and Cox proportional hazards regression analysis was used to evaluate the relationship between SES and prognosis.Results Patients of middle SES had lower unadjusted all-cause recurrence than those of low or high SES;24-week overall recovery rate was 90.4%[95%confidence interval(CI):89.6%–91.2%]in patients of middle SES,13.5%(95%CI:12.8%–14.2%)in patients of low SES,and 31.7%(95%CI:30.7%–32.7%)in patients of high SES(both log-rank P<0.001).After adjustment for covariates,hazard ratios(HRs)were7.69(95%CI:6.17–9.71,Ptrend<0.001)for all-cause recurrence for low SES versus middle SES,and 6.19(95%CI:4.78–7.93,Ptrend<0.001)for middle SES versus high SES.Conclusion Low SES and high SES were more associated with the worse prognosis of CRS patients after ESS than middle SES.

STUDIES ON PATHOGENESIS OF WAARDENBURG SYNDROME TYPE Ⅱ AND TIETZ SYNDROME RESULTING FROM MITF GENE MUTATIONS

Microphthalmia-associated transcription factor (MITF) controls melanocyte survival and differentiation through directly regulating the expression of the tyrosinase (TYR) and tyrosinase-related proteins 1 and 2 (TYRP1 and TYRP2) genes. MITF mutations have been reported to result in an abnormal melanocyte devel-opment and lead to Waardenburg syndrome type 2 (WS2), characterized by variable degrees of sensorineu-ral hearing loss and patchy regional distribution of hypopigmentation. Recently, MITF was also indicated as a causative gene for a more severe syndrome, the Tietz Syndrome (TS), characterized by generalized hy-popigmentation and complete hearing loss. However, few functional studies have been performed to com-pare the diseases-causing mutations. Here, we analyzed the in vitro activity of two recent identified WS2-as-sociated mutation (p.R217I and p.T192fsX18) and one TS-associated mutation p.N210K. The R217I MITF retained partial activity, normal DNA-binding ability and nuclear distribution, whereas the T192fsX18 MITF failed to activate TYR promoter due to loss of DNA-binding activity, and aberrant subcellular localization. The aberrant subcellular localization of T192fsX18 MITF may be caused by deletion of a putative nuclear localization signal (NLS) at aa 213-218 (ERRRRF). Indeed, MITF with deletion of the NLS fragment failed to translocate into the nucleus and activated the TYR promoter. Tagging this NLS to GFP promoted the green fluorescence protein (GFP) translocated into the nucleus. The surprising finding of our study is that a TS-as-sociated MITF mutation, N210K, showed comparable in vitro activity as WT. Thus, the possible involve-ment of MITF in TS and its underlying mechanisms still need further investigation.

TopoisomeraseⅡαGene as a Marker for Prognostic Prediction of Hepatocellular Carcinoma:A Bioinformatics Analysis

Objective To investigate the expression of topoisomeraseⅡα(TOP2α)in hepatocellular carcinoma(HCC)and its role in predicting prognosis of HCC patients.Methods We used HCC-related datasets in UALCAN,HCCDB,and cBioPortal databases to analyze the expression and mutation of TOP2αand its co-expressed genes in HCC tissues.GO function and KEGG pathway enrichment of TOP2αand its co-expressed genes were identified.The TIMER database was used to analyze infiltration levels of immune cells in HCC.The impacts of TOP2αand its co-expression genes and the infiltrated immune cells on the survival of HCC patients were assayed by Kaplan-Meier plotter analysis.Results TOP2αand its co-expression genes were highly expressed in HCC(P<0.001)and detrimental to overall survival of HCC patients(P<0.001).TOP2αand its co-expression genes were mainly involved in cell mitosis and proliferation,and cell cycle pathway(ID:hsa04110,P=0.001945).TOP2αand its co-expression genes were mutated in HCC and the mutations were significantly detrimental to overall survival(P=0.0247)and disease-free survival(P=0.0265)of HCC patients.High TOP2αexpression was positively correlated with the infiltration of B cell(r=0.459,P<0.01),CD8^(+)T cell(r=0.312,P<0.01),CD4^(+)T cell(r=0.370,P<0.01),macrophage(r=0.459,P<0.01),neutrophil(r=0.405,P<0.01),and dendritic cell(r=0.473,P<0.01)in HCC.The CD8^(+)T cell infiltration significantly prolonged the 3-and 5-year survival of HCC patients(all P<0.05),and CD4^(+)T cell infiltration significantly shortened the 3-,5-,and 10-year survival of HCC patients(all P<0.05).Conclusion TOP2αmay be an oncogene,which was associated with poor prognosis of HCC patients and could be used as a biomarker for the prognostic prediction of HCC.

The Comparative and Functional Study between Two Construction Methods of shRNA Expression Vector Targeted LMP1 Gene Encoded by EBV

To look for a more stable and convenient way of constructing short hairpin RNA expression vectors targeting the latent membrane protein-1(LMP-1)encoded by Epstein-Barr virus(pshLMP1),and to study the inhibition function of pshLMP1 expression vectors in HNE1 cells,we designed the pshLMP1 expression cassette and pshLMP1 expression vectors by both the annealing method and PCR method and then co-transfected with pEGFP-N1-1158 into HNE1 cells to observe the mRNA and protein levels of LMP-1 genes by green fluorescence analysis,RT-PCR and western blot.pshLMP1 expression vectors were successfully obtained by both methods but better cloning efficiency was achieved and fewer deletions and mutations of nucleotides were achieved with the PCR method.Furthermore,the mRNA and protein levels of LMP-1 genes were down-regulated by pshLMP1 expression vectors.According to our research,we found that the PCR method provides a more efficient way to construct pshLMP1 expression vectors which have the ability to inhibit the function of LMP-1 genes expressed in HNE1 cells,and also provides a novel application of RNA interference technology against-EBV.

Lentivirus carrying the Atoh1 gene infects normal rat cochlea

Lentivirus carrying the Atohl gene can infect Corti＇s organ and express a hair-like cell surface marker in the supporting cell area. However, expression of the gene carried by adenovirus is instantaneous, which undoubtedly limits its clinical application. Lentivirus acts as a carrier that can stably and continuously express genes. In this study, the cochlear structure and hearing level were not affected, and Atohl gene carried by lentivirus promoted the production of hair-like cells in the cochlear supporting cell area. This led to expression of the hair-like cell surface marker myosin 7a 30 days after lentivirus carrying Atohl was microinjected into the cochlear round window of rats.

Autonomic responses to blast overpressure can be elicited by exclusively exposing the ear in rats

Blast overpressure has become an increasing cause of brain injuries in both military and civilian populations. Though blast＇s direct effects on the cochlea and vestibular organs are active areas of study, little attention has been given to the ear＇s contribution to the overall spectrum of blast injury. Acute auto- nomic responses to blast exposure, including bradycardia and hypotension, can cause hypoxia and contribute to blast-induced neurotrauma. Existing literature suggests that these autonomic responses are elicited through blast impacting the thorax and lungs. We hypothesize that the unprotected ear also provides a vulnerable locus for blast to cause autonomic responses. We designed a blast generator that delivers controlled overpressure waves into the ear canal without impacting surrounding tissues in order to study the ear＇s specific contribution to blast injury. Anesthetized adult rats＇ left ears were exposed to a single blast wave ranging from 0 to 110 PSI （0-758 kPa）. Blast exposed rats exhibited decreased heart rates and blood pressures with increased blast intensity, similar to results gathered using shock tubes and whole-body exposure in the literature. While rats exposed to blasts below 50 PSI （345 kPa） exhibited increased respiratory rate with increased blast intensity, some rats exposed to blasts higher than 50 PSI （345 kPa） stopped breathing immediately and ultimately died. These autonomic responses were significantly reduced in vagally denervated rats, again similar to whole-body exposure literature. These results support the hypothesis that the unprotected ear contributes to the autonomic responses to blast.

Exposure to blast shock waves via the ear canal induces deficits in vestibular afferent function in rats

The ears are air-filled structures that are directly impacted during blast exposure.In addition to hearing loss and tinnitus,blast victims often complain of vertigo,dizziness and unsteady posture,suggesting that blast exposure induces damage to the vestibular end organs in the inner ear.However,the underlying mechanisms remain to be elucidated.In this report,single vestibular afferent activity and the vestibuloocular reflex(VOR)were investigated before and after exposure to blast shock waves(~20 PSI)delivered into the left external ear canals of anesthetized rats.Single vestibular afferent activity was recorded from the superior branch of the left vestibular nerves of the blast-treated and control rats one day after blast exposure.Blast exposure reduced the spontaneous discharge rates of the otolith and canal afferents.Blast exposure also reduced the sensitivity of irregular canal afferents to sinusoidal head rotation at 0.5e2Hz.Blast exposure,however,resulted in few changes in the VOR responses to sinusoidal head rotation and translation.To the best of our knowledge,this is the first study that reports blast exposure-induced damage to vestibular afferents in an animal model.These results provide insights that may be helpful in developing biomarkers for early diagnosis of blast-induced vestibular deficits in military and civilian populations.

Traumatic brain injury induced by exposure to blast overpressure via ear canal

Exposure to explosive shockwave often leads to blast-induced traumatic brain injury in military and civilian populations.Unprotected ears are most often damaged following exposure to blasts.Although there is an association between tympanic membrane perforation and TBI in blast exposure victims,little is known about how and to what extent blast energy is transmitted to the central nervous system via the external ear canal.The present study investigated whether exposure to blasts directed through the ear canal causes brain injury in LongEvans rats.Animals were exposed to a single blast(0–30 pounds per square inch(psi))through the ear canal,and brain injury was evaluated by histological and behavioral outcomes at multiple time-points.Blast exposure not only caused tympanic membrane perforation but also produced substantial neuropathological changes in the brain,including increased expression of c-Fos,induction of a profound chronic neuroinflammatory response,and apoptosis of neurons.The blast-induced injury was not limited only to the brainstem most proximal to the source of the blast,but also affected the forebrain including the hippocampus,amygdala and the habenula,which are all involved in cognitive functions.Indeed,the animals exhibited long-term neurological deficits,including signs of anxiety in open field tests 2 months following blast exposure,and impaired learning and memory in an 8-arm maze 12 months following blast exposure.These results suggest that the unprotected ear canal provides a locus for blast waves to cause TBI.This study was approved by the Institutional Animal Care and Use Committee at the University of Mississippi Medical Center(Animal protocol#0932 E,approval date:September 30,2016 and 0932 F,approval date:September 27,2019).

Passive eye movements induced by electromagnetic force(EMF) in rats

Accurate information on eye position in the orbit is available from visual feedback,efference copy of the oculomotor commands and proprioceptive signals from the extraocular muscles (EOM).Whereas visual feedback and oculomotor commands have been extensively studied,central processing of EOM proprioceptive signals remains to be elucidated.A challenge to the field is to develop an approach to induce passive eye movements without physically contacting the eyes.A novel method was developed to generate passive eye movements in rats.A small rare-earth magnet disk (0.7 mm diameter,0.5 mm thickness) was attached to the surface of a rat's eyeball.A metal rod (5 mm diameter) wrapped with an electromagnetic (EM) coil was placed near the magnet (8-15 mm).By passing currents to the EM coil,electromagnetic force (EMF) was generated and acted upon the magnet and induced passive eye movements.The EMF induced well-defined passive eye movements,whose directions were dependent on current polarity and amplitudes and peak velocities were dependent on current intensity and duration.Peak velocities of the EMF-induced eye movements were linearly related to amplitudes,exhibiting main sequence relationships similar to that of saccades in awake rats and eye movements induced by electrical microstimulation of the abducens nucleus in anesthetized rats.Histological examination showed that repetitive EMF stimulations did not appear to result in damages in the EOM fibers.These results validated the EMF approach as a novel tool to investigate EOM proprioceptive signals and their roles in visual localization and gaze control.

Transnasal endoscopic repair of acquired posterior choanal stenosis and atresia

Background There are congenital and acquired choanal atresias and many approaches have been used for their repair. We assessed the clinical effect of power instrument, endoscopic repair of acquired choanal stenosis and atresia. Methods Nineteen patients, aged from 32 to 61 years, with acquired choanal stenosis and atresia （from trauma in 5 cases and from radiotherapy after nasopharyngeal carcinoma in 14; 6 bilateral and 13 unilateral cases）, underwent transnasal endoscopic repair of choanal stenosis and atresia. No patient had stenting. Antibiotic and local glucocorticoid were administered postoperatively.Results Eighteen patients remained free of symptoms for 12-40 months after the surgery, and the diameter of the neochoana was more than 1 cm after the procedure. One patient required revision surgery and recovered completely with no restenosis at 12 months after the second surgery. There were no postoperative complications. Histology of the resected tissue revealed respiratory epithelial-lined stromal tissue with chronic inflammation, edema and fibrosis, but no tumor cells.Conclusions Transnasal endoscopic approach is a useful procedure for the repair of acquired choanal stenosis and atresia： it is highly successful, safe and effective with swift recovery and short time of hospitalization. It is very important in postoperative care to remove any granulation or polyps at the site of the neochoana at that time.

Giant cell angiofibroma in the vocal cord

Giant cell angiofibroma （GCA） was first described in 1995, and it is a rare tumor in soft tissues,1 Since the initial report of the disease in orbital tissue, GCA has been reported in a variety of other soft tissues.2,3 The morphological hallmark is richly vascularized, patternless spindle cell proliferation in the presence of pseudovascular spaces and multinucleated giant cells. The pathological diagnosis of GCA is mainly based on positive staining of CD34. GCA is usually considered to be a benign tumor. In clinic, GCA needs to be distinguished from other tumors in soft tissues. Herein,we report two rare cases of GCA in the vocal cord and describe its histological features.

IVS8 ＋ 1 DelG, a Novel Splice Site Mutation Causing DFNA5 Deafness in a Chinese Family

Background： Nonsyndrornic hearing loss （NSHL） is highly heterogeneous, in which more than 90 causative genes have currently been identified. DFNA5 is one of the deathess genes that known to cause autosomal dominant NSHL. Until date, only five DFN.45 mutations have been described in eight families worldwide. In this study, we reported the identification of a novel pathogenic mutation causing DFNA5 deafness in a five-generation Chinese family. Methods： Alter detailed clinical evaluations of this family, the genomic DNA of three affected individuals was selected for targeted exome sequencing of 101 known deafness genes, as well as mitochondrial DNA and microRNA regions. Co-segregation analysis between the hearing loss and the candidate variant was confirmed in available family members by direct polymerase chain reaction （PCR）-Sanger sequencing. Real-time PCR （RT-PCR） was pertormed to investigate the potential effect of the pathogenic mutation on messenger RNA splicing. Results： Clinical evaluations revealed a similar deafness phenotype in this family to that of previously reported DFNA5 families with autosomal dominant, late-onset bearing loss. Molecular analysis identified a novel splice site mutation in DFNA5 intron 8 （IVSS＋ 1 delG）. The mutation segregated with the hearing loss of the family and was absent in 120 unrelated control DNA samples of Chinese origin. RT-PCR showed skipping of exon 8 in the mutant transcript. Conclusions： We identified a novel DFNA5 mutation IVS8＋1 delG in a Chinese family which led to skipping ofexon 8. This is the sixth DFNA5 mutation relates to hearing loss and the second one in DFNA5 intron 8. Our findings provide further support to the hypothesis that the DFNA5-associated hearing loss represents a mechanism of gain-of-function.

Gait instability in patients with small acoustic neuroma

Background Small acoustic neuromas seldom result in typical vestibular symptoms, despite the tumor arising from the vestibular nerve. In this study, we have shown that abnormal gait in eleven patients with small acoustic neuroma could be detected in gait analysis by the use of tactile sensor. Patients displayed no oculomotor abnormality and had tumors less than 10 mm from the porus acoustics. Methods Gait related parameters including the coefficients of variations （CV） of stance, swing, double support, area ratio of trajectories of center of force （TCOF）, in addition to the foot pressure difference between both feet, were used for assessment of gait.