[4 + 1 + 1] Tandem Cyclization Reaction Involving Isocyanides: Access to 2-(Trifluoromethyl)quinazolin-4(3H)-imines.

A palladium-catalyzed three-component reaction of isocyanides, 2,2,2-trifluoro-N-(2-iodophenyl)acetimidoyl chlorides, and amines for the one-pot synthesis of 2-(trifluoromethyl)quinazolin-4(3H)-imines was described. The protocol features a wide substrate scope, high efficiency, and readily available raw materials.

Recommendations for head and neck surgical oncology practice in a setting of acute severe resource constraint during the COVID-19 pandemic: an international consensus.

The speed and scale of the global COVID-19 pandemic has resulted in unprecedented pressures on health services worldwide, requiring new methods of service delivery during the health crisis. In the setting of severe resource constraint and high risk of infection to patients and clinicians, there is an urgent need to identify consensus statements on head and neck surgical oncology practice. We completed a modified Delphi consensus process of three rounds with 40 international experts in head and neck cancer surgical, radiation, and medical oncology, representing 35 international professional societies and national clinical trial groups. Endorsed by 39 societies and professional bodies, these consensus practice recommendations aim to decrease inconsistency of practice, reduce uncertainty in care, and provide reassurance for clinicians worldwide for head and neck surgical oncology in the context of the COVID-19 pandemic and in the setting of acute severe resource constraint and high risk of infection to patients and staff.

Assessment of a biofluid mechanics-based model for calculating portal pressure in canines.

BACKGROUND: Portal hypertension is a severe complication caused by various chronic liver diseases. The standard methods for detecting portal hypertension (hepatic venous pressure gradient and free portal pressure) are available in only a few hospitals due to their technical difficulty and invasiveness; thus, non-invasive measuring methods are needed. This study aimed to establish and assess a novel model to calculate free portal pressure based on biofluid mechanics. RESULT: Comparison of each dog's virtual and actual free portal pressure showed that a biofluid mechanics-based model could accurately predict free portal pressure (mean difference: -0.220, 95% CI: - 0.738 to 0.298; upper limit of agreement: 2.24, 95% CI: 1.34 to 3.14; lower limit of agreement: -2.68, 95% CI: - 3.58 to - 1.78; intraclass correlation coefficient: 0.98, 95% CI: 0.96 to 0.99; concordance correlation coefficient: 0.97, 95% CI: 0.93 to 0.99) and had a high AUC (0.984, 95% CI: 0.834 to 1.000), sensitivity (92.3, 95% CI: 64.0 to 99.8), specificity (91.7, 95% CI: 61.5 to 99.8), positive likelihood ratio (11.1, 95% CI: 1.7 to 72.8), and low negative likelihood ratio (0.08, 95% CI: 0.01 to 0.6) for detecting portal hypertension. CONCLUSIONS: Our study suggests that the biofluid mechanics-based model was able to accurately predict free portal pressure and detect portal hypertension in canines. With further research and validation, this model might be applicable for calculating human portal pressure, detecting portal hypertensive patients, and evaluating disease progression and treatment efficacy.

The discovery of a novel eight-mRNA-lncRNA signature predicting survival of hepatocellular carcinoma patients.

Increasing evidence indicates that the expressions of messenger RNAs (mRNAs) and long non-coding RNAs (lncRNAs) undergo a frequent and aberrant change in carcinogenesis and cancer development. But some research was carried out on mRNA-lncRNA signatures for prediction of hepatocellular carcinoma (HCC) prognosis. We aimed to establish an mRNA-lncRNA signature to improve the ability to predict HCC patients' survival. The subjects from the cancer genome atlas (TCGA) data set were randomly divided into two parts: training data set (n = 246) and testing data set (n = 124). Using computational methods, we selected eight gene signatures (five mRNAs and three lncRNAs) to generate the risk score model, which were significantly correlated with overall survival of patients with HCC in both training and testing data set. The signature had the ability to classify the patients in training data set into a high-risk group and low-risk group with significantly different overall survival (hazard ratio = 4.157, 95% confidence interval = 2.648-6.526, P < 0.001). The prognostic value was further validated in testing data set and the entire data set. Further analysis revealed that this signature was independent of tumor stage. In addition, Gene Set Enrichment Analysis suggested that high risk score group was associated with cell proliferation and division related pathways. Finally, we developed a well-performed nomogram integrating the prognostic signature and other clinical information to predict 3- and 5-year overall survival. In conclusion, the prognostic mRNAs and lncRNAs identified in our study indicate their potential role in HCC biogenesis. The risk score model based on the mRNA-lncRNA may be an efficient classification tool to evaluate the prognosis of patients' with HCC.

Palladium Catalyzed Insertion Reaction of Isocyanides with 3-Arylisoxazol-5(4H)-ones: Synthesis of 4-Aminomethylidene Isoxazolone Derivates.

A palladium catalyzed insert reaction of isocyanides to 3-arylisoxazol-5(4H)-ones for the construction of 4-aminomethylidene isoxazolone derivates is reported. In this transformation, only the C-H bond of the methylene group was involved while the remaining ring structure was retained. In general, this work provided a new protocol for the synthesis of 4-aminomethylidene isoxazolones.

Identification of potential diagnostic biomarkers of acute pancreatitis by serum metabolomic profiles.

Acute pancreatitis (AP) is defined as an acute inflammation of pancreas that may cause damage to other tissues and organs depending upon the severity of symptoms. The diagnosis of AP is usually made by detection of raised circulating pancreatic enzyme levels, but there are occasional false positive and false negative diagnoses and such tests are often normal in delayed presentations. More accurate biomarkers would help in such situations. In this study, the global metabolites' changes of AP patients (APP) were profiled by using gas chromatography-mass spectrometry (GC-MS). Multivariate pattern recognition techniques were used to establish the classification models to distinguish APP from healthy participants (HP). Some significant metabolites including 3-hydroxybutyric acid, phosphoric acid, glycerol, citric acid, d-galactose, d-mannose, d-glucose, hexadecanoic acid and serotonin were selected as potential biomarkers for helping clinical diagnosis of AP. Furthermore, the metabolite changes in APP with severe and mild symptoms were also analyzed. Based on the selected biomarkers, some relevant pathways were also identified. Our results suggested that GC-MS based serum metabolomics method can be used in the clinical diagnosis of AP by profiling potential biomarkers.

Arthroscopic Latarjet Procedure With Anterior Capsular Reconstruction: Clinical Outcome and Radiologic Evaluation With a Minimum 2-Year Follow-Up.

PURPOSE: To investigate the clinical and radiographic outcomes of the modified arthroscopic Latarjet procedure at a minimum of 2 years after surgery. METHODS: Patients who had traumatic unidirectional anterior shoulder instability and treated with a modified arthroscopic Latarjet procedure were included. During surgery, the anterior capsule was preserved and repaired back to the glenoid after the coracoid transfer. The clinical results (range of motion, American Shoulder and Elbow Surgeons [ASES] score, Constant-Murley score, and Rowe score) and computed tomographic results were followed. RESULTS: From February 2013 to September 2014, 52 consecutive patients were included. The average duration of follow-up was 28.4 months (range, 24.0-41.7 months). At final follow-up, no recurrent dislocation had occurred. The ASES score and Rowe score improved significantly (ASES score from 85.6 ± 12.7 before surgery to 93.6 ± 5.4 after surgery, P < .0001; Rowe score from 41.5 ± 7.2 before surgery to 92.2 ± 8.7 after surgery, P < .0001). No significant change was found regarding range of motion and the Constant-Murley score. Bone union was achieved in all cases. The transferred coracoid was at the level of the glenoid in all cases. The transferred coracoid was placed below the equator in 48 of 52 cases (92.3%). The orientation of the screw was 22.6° ± 10.8°. Bone resorption around the proximal screw was significantly more prominent than that around the distal screw (P < .0001). CONCLUSIONS: The arthroscopic Latarjet procedure with concomitant anterior capsular reconstruction can achieve satisfactory clinical outcomes for the treatment of anterior shoulder instability with marked glenoid bone loss at a minimum of 2 years' follow-up. A satisfactory coracoid graft position, proper screw orientation, and high healing rate of the transferred coracoid can be expected. Bone resorption around the proximal screw is more severe than that around the distal screw. LEVEL OF EVIDENCE: Level IV, therapeutic case series.

Coracoid bone graft resorption after Latarjet procedure is underestimated: a new classification system and a clinical review with computed tomography evaluation.

BACKGROUND: This study proposes a simple and reliable classification system to evaluate the severity of the bone resorption of the transferred coracoid bone block after the Latarjet procedure. The incidence and severity of the graft resorption was also investigated. METHODS: Between January 2009 and January 2012, 63 patients underwent an open Latarjet procedure and were included. Four independent surgeons used the classification system we proposed to evaluate the severity of the graft resorption on the computed tomography scan performed 1 year postoperatively. Each surgeon did the evaluation twice at a 3-month interval. The interobserver and intraobserver reliability of the classification system were analyzed using intraclass correlation coefficients. Among these 63 patients, 57 patients were available for clinical evaluation at 2 years postoperatively. RESULTS: The American Shoulder and Elbow Surgeons score, Constant-Murley score, and Rowe score were improved significantly after the surgery. No redislocation occurred during follow-up. The incidence of graft resorption was 90.5% based on the computed tomography evaluation. The coracoid graft resorption was classified as grade 0 in 6 patients, grade I in 26, grade II in 25, and grade III in 6. The classification system had excellent interobserver and intraobserver reliability. CONCLUSION: The open Latarjet procedure is effective in treating anterior shoulder instability with marked glenoid bone loss. The incidence of the graft resorption at 1 year postoperatively is high. Our classification system on the graft resorption after Latarjet procedure has good interobserver and intraobserver reliability.

Comparison of complications of the suprameatal approach and mastoidectomy with posterior tympanotomy approach in cochlear implantation: a meta-analysis.

BACKGROUND: Cochlear implantation (CI) is a popular procedure to preserve hearing in patients with severe-to-profound hearing loss. Evidence shows that the suprameatal approach (SMA) may help reducing the risk of the incidence of complications and shortening the surgery time, but there is still dispute. OBJECTIVES: The aim of this study was to compare the incidence of complications of SMA and the mastoidectomy with posterior tympanotomy approach (MPTA), and to find whether SMA yields better outcomes than MPTA. METHODS: We searched PubMed, the Cochrane Library, the Web of Science and Chinese Biomedical Literature databases, Chinese National Knowledge Infrastructure, the Chinese Science and Technology Journal Full-Text database, and Wangfang database. The latest data was accessed in March 2013. Review Manager 5.1 software was used for comprehensive quantification data analysis. RESULTS: Three studies were included in the meta-analysis, composed of 799 participants and reporting major and minor complications. The meta-analysis indicated no statistically significant difference in major and minor complications between the two approaches, except for facial nerve and chorda tympani injuries (OR = 0.13; 95% CI: 0.02, 0.67; p = 0.02; I(2) = 0%). CONCLUSIONS: Current evidence suggests that SMA may be clearly a good alternative to the classical surgery technique for CI in terms of reducing the incidence of facial nerve injury and chorda tympani sacrifice.

A novel intronic variant causing aberrant splicing identified in two deaf Chinese siblings with enlarged vestibular aqueducts.

OBJECTIVE: We aimed to evaluate the genotype-phenotype relationship in two Chinese family members with enlarged vestibular aqueduct (EVA). METHODS: We collected blood samples and clinical data from each pedigree family member. Genomic DNA was isolated from peripheral leukocytes using standard methods. Targeted next-generation sequencing and Sanger sequencing were performed to find the pathogenic mutation in this family. Minigene assays were used to verify whether the novel intronic mutation SLC26A4c.765+4A>G influenced mRNA splicing. RESULTS: Hearing loss in the patients with EVA was diagnosed using auditory tests and imaging examinations. Two pathogenic mutations, c.765+4A>G and c.919-2A>G were detected in SLC26A4. In vitro minigene analysis confirmed that c.765+4A>G variant could cause aberrant splicing, resulting in skipping over exon 6. CONCLUSIONS: The SLC26A4c.765+4A>G mutation is the causative variant in the Chinese family with EVA. Particular attention should be paid to intronic variants.

Divergent Synthesis of Pentacyclic Isoindolinones Enabled by Sequential Insertion of Two Different Isocyanides and Acid Promoted Cyclization of Ketenimines.

A palladium-catalyzed multicomponent reaction involving o-bromobenzaldehydes and two different isocyanides was developed to assemble series of isoindolinones with spiroindolenine or azepinoindole skeletons. This sequential insertion reaction features mild conditions, a wide substrate scope, and high efficiency. Preliminary mechanistic study indicated that the difference in steric hindrance between isocyanide components is crucial when regulating the reaction sequence, whereas the ligand also played an important role during the whole process.

Integrated approaches for increasing plant yield under salt stress.

Salt stress affects large cultivated areas worldwide, thus causing remarkable reductions in plant growth and yield. To reduce the negative effects of salt stress on plant growth and yield, plant hormones, nutrient absorption, and utilization, as well as developing salt-tolerant varieties and enhancing their morpho-physiological activities, are some integrative approaches to coping with the increasing incidence of salt stress. Numerous studies have been conducted to investigate the critical impacts of these integrative approaches on plant growth and yield. However, a comprehensive review of these integrative approaches, that regulate plant growth and yield under salt stress, is still in its early stages. The review focused on the major issues of nutrient absorption and utilization by plants, as well as the development of salt tolerance varieties under salt stress. In addition, we explained the effects of these integrative approaches on the crop's growth and yield, illustrated the roles that phytohormones play in improving morpho-physiological activities, and identified some relevant genes involve in these integrative approaches when the plant is subjected to salt stress. The current review demonstrated that HA with K enhance plant morpho-physiological activities and soil properties. In addition, NRT and NPF genes family enhance nutrients uptake, NHX1, SOS1, TaNHX, AtNHX1, KDML, RD6, and SKC1, maintain ion homeostasis and membrane integrity to cope with the adverse effects of salt stress, and sd1/Rht1, AtNHX1, BnaMAX1s, ipal-1D, and sft improve the plant growth and yield in different plants. The primary purpose of this investigation is to provide a comprehensive review of the performance of various strategies under salt stress, which might assist in further interpreting the mechanisms that plants use to regulate plant growth and yield under salt stress.

Effects of gradient high-field static magnetic fields on diabetic mice.

Although 9.4 T magnetic resonance imaging (MRI) has been tested in healthy volunteers, its safety in diabetic patients is unclear. Furthermore, the effects of high static magnetic fields (SMFs), especially gradient vs. uniform fields, have not been investigated in diabetics. Here, we investigated the consequences of exposure to 1.0-9.4 T high SMFs of different gradients (>10 T/m vs. 0-10 T/m) on type 1 diabetic (T1D) and type 2 diabetic (T2D) mice. We found that 14 h of prolonged treatment of gradient (as high as 55.5 T/m) high SMFs (1.0-8.6 T) had negative effects on T1D and T2D mice, including spleen, hepatic, and renal tissue impairment and elevated glycosylated serum protein, blood glucose, inflammation, and anxiety, while 9.4 T quasi-uniform SMFs at 0-10 T/m did not induce the same effects. In regular T1D mice (blood glucose ≥16.7 mmol/L), the >10 T/m gradient high SMFs increased malondialdehyde ( P<0.01) and decreased superoxide dismutase ( P<0.05). However, in the severe T1D mice (blood glucose ≥30.0 mmol/L), the >10 T/m gradient high SMFs significantly increased tissue damage and reduced survival rate. In vitro cellular studies showed that gradient high SMFs increased cellular reactive oxygen species and apoptosis and reduced MS-1 cell number and proliferation. Therefore, this study showed that prolonged exposure to high-field (1.0-8.6 T) >10 T/m gradient SMFs (35-1 380 times higher than that of current clinical MRI) can have negative effects on diabetic mice, especially mice with severe T1D, whereas 9.4 T high SMFs at 0-10 T/m did not produce the same effects, providing important information for the future development and clinical application of SMFs, especially high-field MRI.

[Primary shoulder arthroplasty for treatment of complex proximal humeral fractures with the trabecular metal prosthesis].

OBJECTIVE: To evaluate the clinical results of primary shoulder arthroplasty for the treatment of complex proximal humeral fractures with the trabecular metal prosthesis. METHODS: In the study, 57 consecutive patients with complex proximal humeral fractures underwent primary shoulder arthroplasty with the trabecular metal prosthesis, of whom 45 patients were available for the final follow-up (78.9%), and were retrospectively assessed. There were 15 men and 30 women with the average age of (64.9±10.6) years (range: 45 to 85 years), and 14 left shoulders and 31 right shoulders were involved. The mean time from injury to surgery was (10.2±4.8) days. According to Neer's classification, there were 3 patients with a three-part fracture, 4 with a three-part dislocation, 2 with a four-part fracture, 22 with a four-part dislocation, 4 with a four-part valgus-impaction, and 10 with a head splitting. Thirty-nine patients were treated with hemiarthroplasty, and the rest with total shoulder arthroplasty. RESULTS: After a mean follow-up time of (18.7±8.1) months (range: 12 to 41 months), the average range of motion of patients were 130.4°±36.5° for forward elevation, 37.6°±16.6° for external rotation and L3 level (±3 vertebrae) for internal rotation. The average VAS pain score, ASES score, Constant-Murley score and UCLA score were 0.5±1.2 (0-4), 81.4±13.9 (43-100), 81.6±13.6 (52-100) and 28.5±5.1 (16-35), respectively. The average patient satisfaction rate was 95.6%. The greater tuberosity healed anatomically in 42 patients, which was confirmed by the postoperative radiographs, while the greater tuberosity nonunion was found in the other three patients. Superior migration of humeral head was found in two patients. There were no complications, such as infection, prosthetic loosening or neurovascular injury, related to the surgery. CONCLUSION: Primary shoulder arthroplasty for complex proximal humeral fractures with the trabecular metal prosthesis could yield good clinical results. The healing rate of the greater tuberosity was 93.3%.

[Preliminary histological and biomechanical study about the timing of surgical repair for acute rotator cuff tears in rabbits].

OBJECTIVES: To establish the animal model of acute rotator cuff tear in rabbits, and study the effect of timing of surgical repair on healing of tendon-bone interface, formation and distribution of collagens in the supraspinatus tendon insertion and biomechanical properties of supraspinatus. METHODS: Supraspinatus tenotomy was performed in the right shoulder of 90 skeletally matured male New Zealand white rabbits to establish the animal model of acute rotator cuff tear. The rabbits were randomly divided into 3 groups : group of early repair, repaired at 1 week after tenotomy; group of late repair, repaired at 4 weeks after tenotomy; and group without repair, used as control. At 2 weeks, 4 weeks and 8 weeks after repair, healing of tendon-bone interface was observed by HE staining. Collagens were observed by Sirius Red F 3B (SR) in saturated carbazotic acid staining. The areas of type I and III collagens were measured by using imaging analysis software and the ratio of type I and III collagens were calculated. Failure loads of supraspinatus on both sides were measured. The percentage of failure loads of the surgical side was calculated and contralateral supraspinatus were uninjured. RESULTS: There was no obvious fatty infiltration and muscle atrophy in supraspinatus in all groups. At 8 weeks, the formation of a new enthesis of supraspinatus in groups of early and late repair were observed. In groups of early and late repair, the ratio of areas of type I and III collagens at 8 weeks (2.02 ± 0.77 and 2.06 ± 0.58) was larger than that at 2 weeks (1.10 ± 0.24 and 1.14 ± 0.50, t = 3.082, 3.655, P < 0.01). At 2, 4 and 8 weeks, the percentages of failure loads of the surgical side and uninjured contralateral supraspinatus in group of early repair(38% ± 11%, 66% ± 7%, 89% ± 4%) and group of late repair (41% ± 16%, 63% ± 7%, 89% ± 9%) were both higher than that in group without repair (14% ± 6%, 32% ± 4%, 56% ± 12%); the differences were all statistically significant (group of early repair: t = 3.311, 8.549, 5.719; group of late repair: t = 3.713, 8.063, 6.044; P < 0.01). The percentage of failure loads of the surgical side and uninjured contralateral supraspinatus at 8 weeks was higher than those at 4 weeks (t = 3.878 - 4.613, P < 0.01) and 2 weeks (t = 7.158 - 10.024, P < 0.01) in all groups. CONCLUSIONS: Surgical repair within 4 weeks of acute rotator cuff tear lead to formation of a new enthesis of supraspinatus, improvement of both ratio of type I collagen in the supraspinatus tendon insertion and biomechanical properties of supraspinatus.

[Clinical follow-up study of combined tenodesis for long head of biceps tendon lesion with massive rotator cuff tear].

OBJECTIVE: To evaluate the combined tenodesis for long head of bicep tendon lesion with massive rotator cuff tear. METHODS: From January 2004 to June 2009, 41 patients of long head of bicep tendon lesion with massive rotator cuff tear were treated by arthroscopy. The follow-up period was over 12 months. There were 19 males and 22 females with an average age of 57 ± 11 year old. All patients were treated by arthroscopy with double row technique for rotator cuff tear repair and tenodesis for lesions of long head of bicep tendon. It combined the long head of bicep tendon with anterior edge of rotator cuff. The mean visual analogue scale (VAS) was 5.1 ± 23, the range of forward flexion (100 ± 55)°, external rotation (27 ± 24)° and internal rotation up to T(12) (T(5)-S(1)) level pre-operation on average. The Constant-Merly score was 50 ± 23, the University of California at Los Angeles (UCLA) score 14 ± 6 and the simple shoulder test (SST) score 4.0 ± 2.7 pre-operation on average. The strength of flexed elevation was (17 ± 20)% and strength of elbow flexion (101 ± 16)% versus the other side. RESULTS: All patients healed without any complication. Their outcomes improved significantly (P < 0.01). The mean VAS score improved to 1.5 ± 1.8, forward flexion (155 ± 21)°, external rotation (37 ± 24)° and post-operative internal rotation up to T(10) (T(4)-L(3)) on average. The mean Constant improved to 87 ± 13, mean UCLA 29 ± 5 and mean SST 9.3 ± 2.4. The strength of flexed elevation recovered to (68 ± 21)% post-operation. Significant differences were found in range of motion, VAS, strength and functional score (P < 0.01). No difference was observed for Mayo elbow performance score (MEPS) and elbow flexion strength at pre-operation versus post-operation. CONCLUSION: The combined tenodesis with double row rotator cuff repair is an effective approach for treating lesion of long head of bicep tendon with massive rotator cuff tear.

[Clinical results after all arthroscopic reduction and fixation of bony Bankart lesion].

OBJECTIVE: To investigate the shoulder function after arthroscopic reduction and internal fixation in patients with bony Bankart lesion. METHODS: Between May 2004 and May 2008, 45 patients with bony Bankart lesion who were treated with all arthroscopic reduction and internal fixation with metal anchors were included in this study. Among them 40 patients were male and 5 patients were female. The average age at the surgery was 27.6 years (16.5 - 50.1 years). The average duration of follow-up was 29.7 months (24.8 - 49.0 months). A history of recurrent dislocation of affected shoulder was found in all patients. Metal anchors were used to fix the bony Bankart lesion during the surgery. Hill-Sachs remplissage technique was used to treat the Engaging Hill-Sachs lesion. The preoperative American Shoulder and Elbow Society (ASES) score, Constant-Murley score, Rowe score and the VAS score for instability were 84 ± 14, 95.1 ± 4.6, 39.4 ± 2.9 and 5 ± 3 respectively. RESULTS: No significant change was found regarding active forward elevation, external rotation and internal rotation after the surgery. The ASES score, Constant-Murley score, Rowe score and the VAS score of stability were 95 ± 7, 98.3 ± 2.2, 84.5 ± 22.0 and 1 ± 2, improved significantly higher after the surgery (P < 0.01). One subluxation and 3 redislocation were happened during follow-up. The overall failure rate was 8.9% (4/45). CONCLUSIONS: All arthroscopic reduction and fixation of bony Bankart lesion can achieve a good result.

Divergent Reaction of Isocyanides with o-Bromobenzaldehydes: Synthesis of Ketenimines and Lactams with Isoindolinone Cores.

A divergent reaction of isocyanides with o-bromobenzaldehydes for the synthesis of isoindolinone-derived ketenimines and lactams was disclosed. The reaction features readily available reactants, relatively mild conditions, and high yields of products. Ketenimines could be applied in further transformations for access to other functional molecules. A mechanism study showed that the palladium-migration/imine-insertion process was the key step in this reaction.

Familial Temperature-Sensitive Auditory Neuropathy: Distinctive Clinical Courses Caused by Variants of the OTOF Gene.

Objective: To investigate the clinical course and genetic etiology of familial temperature-sensitive auditory neuropathy (TSAN), which is a very rare subtype of auditory neuropathy (AN) that involves an elevation of hearing thresholds due to an increase in the core body temperature, and to evaluate the genotype-phenotype correlations in a family with TSAN. Methods: Six members of a non-consanguineous Chinese family, including four siblings complaining of communication difficulties when febrile, were enrolled in this study. The clinical and audiological profiles of the four siblings were fully evaluated during both febrile and afebrile episodes, and the genetic etiology of hearing loss (HL) was explored using next-generation sequencing (NGS) technology. Their parents, who had no complaints of fluctuating HL due to body temperature variation, were enrolled for the genetics portion only. Results: Audiological tests during the patients' febrile episodes met the classical diagnostic criteria for AN, including mild HL, poor speech discrimination, preserved cochlear microphonics (CMs), and absent auditory brainstem responses (ABRs). Importantly, unlike the pattern observed in previously reported cases of TSAN, the ABRs and electrocochleography (ECochG) signals of our patients improved to normal during afebrile periods. Genetic analysis identified a compound heterozygous variant of the OTOF gene (which encodes the otoferlin protein), including one previously reported pathogenic variant, c.5098G > C (p.Glu1700Gln), and one novel variant, c.4882C > A (p.Pro1628Thr). Neither of the identified variants affected the C2 domains related to the main function of otoferlin. Both variants faithfully cosegregated with TSAN within the pedigree, suggesting that OTOF is the causative gene of the autosomal recessive trait segregation in this family. Conclusion: The presence of CMs with absent (or markedly abnormal) ABRs is a reliable criterion for diagnosing AN. The severity of the phenotype caused by dysfunctional neurotransmitter release in TSAN may reflect variants that alter the C2 domains of otoferlin. The observations from this study enrich the current understanding of the phenotype and genotype of TSAN and may lay a foundation for further research on its pathogenesis.

Targeted next-generation sequencing identifies a novel frameshift EYA1 variant causing branchio-otic syndrome in a Chinese family.

OBJECTIVE: To evaluate the genotype-phenotype correlation of branchio-otic syndrome (BOS) in a Chinese family. METHODS: The proband in this study was an 18-month-old boy with hearing loss, preauricular pit, and branchial fistula without a renal anomaly. We collected blood samples from 6 family members, including 4 who were affected by the syndrome. Targeted next-generation sequencing and Sanger sequencing were performed to identify pathogenic mutations in this family. RESULTS: Pedigree analysis indicated that the mode of inheritance in the family was consistent with the autosomal dominant pattern. Hearing loss was the most common manifestation, occurring in 4 patients. Other findings included preauricular pits (n=2), cervical fistulas (n=3) and abnormal pinnae (n=4). None of the patients had renal anomalies. Evaluation by pure-tone audiometry and temporal bone imaging demonstrated bilateral mixed hearing loss, as well as middle ear and inner ear deformities, in two patients. Mutational analysis of candidate genes in the selected patients led to the identification of a novel frameshift variant NM_000503.4: c.1075_1077delinsAT (p.Gly359Ilefs*7) in the EYA1 gene. CONCLUSIONS: The EYA1 c.1075_1077delinsAT mutation is the causative variant in the Chinese family with BOS, although the penetrance is variable within patients.