OBJECTIVE:To characterize the clinical features, management, outcomes, and predictors of recurrence of cerebrospinal fluid otorrhea in patients with inner ear malformations. METHODS:This retrospective cohort study included 40 patients with inner ear malformation-associated cerebrospinal fluid otorrhea who underwent surgical repair at our institution between January 2009 and December 2024. Data on patient demographics, clinical presentation, imaging findings, surgical management, and outcomes were collected and analyzed. RESULTS:The cohort predominantly comprised minors (97.5%, 39/40), with a mean age of onset of 5.83 years. Young children (≤7 y) accounted for 80% (32/40) of patients. Meningitis was the most common presenting symptom (75%), and the overall incidence throughout the disease course was 97.5% (39/40). Recurrent meningitis was observed in 57.5% (23/40) of patients preoperatively, with the frequency of episodes significantly correlating with the duration of the preoperative interval. Radiologically, an incomplete partition type I (IP-I) malformation was the most prevalent inner ear anomaly, and the cerebrospinal fluid (CSF) leakage sites were almost exclusively localized to the oval window region. The initial surgical success rate was 77.5% (31/40), with most recurrences occurring within 2 years postoperatively. Causes of recurrence included reperforation at the primary leak site, misdiagnosis, missed diagnosis, and failure to address the oval window region. CONCLUSIONS:Cerebrospinal fluid otorrhea associated with inner ear malformations primarily afflicts the pediatric population and most commonly presents as meningitis. Early and accurate diagnosis is critical for definitive management. The selection of an optimal surgical approach and meticulous repair technique are pivotal for achieving a high cure rate and minimizing the risks of failure or recurrence.

OBJECTIVE:This study aimed to investigate the spectrum of deafness gene variants among newborns in Hainan Province, China. We also evaluated the effectiveness of a combined hearing and genetic screening approach in this region. Special attention was given to differences between the Han and Li populations, as well as the relationship between genetic variants and hearing follow-up outcomes. METHODS:From October 2020 to June 2022, 2128 newborns in Hainan (1502 Han, 626 Li) received combined hearing and genetic screening. Hearing screening followed national standards. Genetic screening was carried out using targeted next-generation sequencing of GJB2, SLC26A4, GJB3, and MT-RNR1. Infants with detected variants or those referred from hearing screening received diagnostic audiological follow-up. RESULTS:Among the 2128 newborns (1502 Han, 626 Li), 514 (24.15%) carried deafness gene variants. Variants in GJB2 were the most common, with a carrier rate of 22.51% (479/2128). The GJB2 c.109G > A (p.V37I) variant was the hotspot variant, with an overall carrier rate of 20.91% (445/2128). This rate was significantly higher in the Li ethnic group (28.6%, 179/626) than in the Han group (17.7%, 266/1502; P < 0.01). Thirty-seven newborns carried biallelic pathogenic variants in GJB2. Nine newborns (0.42%) were diagnosed with hearing loss at three months, and all 9 had biallelic GJB2 variants (7 homozygous c.109G > A, 1 compound heterozygote c.109G > A/299_300del, and 1 homozygous c.235del). Longitudinal follow-up of children with biallelic c.109G > A variants showed delayed-onset and progressive hearing loss, with confirmed cases increasing at ages 2 and 4. There are no significant differences in mean ABR thresholds or their standard deviations between Han and Li individuals with homozygous GJB2 c.109G > A mutations at 4 years of age. For patients with biallelic c.109G > A variants, the current setting of ABR thresholds may lead to missed diagnosis. CONCLUSION:This study showed a high prevalence of deafness gene variants, especially GJB2 c.109G > A, among newborns in Hainan, with clear ethnic differences. Combined screening was effective in identifying genetic hearing loss, including delayed-onset and progressive cases that could be missed by hearing screening alone. These findings support adding genetic screening to regional newborn screening programs and highlight the need for long-term monitoring of at-risk children.

OBJECTIVE:This study aimed to characterize spontaneous CSF otorrhea or otorhinorrhea from skull base defects, including its clinical presentation, management outcomes, and causes of recurrence. METHODS:A retrospective cohort study was performed on 20 patients who underwent surgical repair at Chinese PLA General Hospital between 2009 and 2024. RESULTS:The cohort was predominantly adult (75%), male, and right-sided. Key presentations were conductive hearing loss (85%), otorrhea (50%), and meningitis (35%). Most defects (85%) involved the middle cranial fossa floor. Initial surgical success was 75%, with recurrence (mean 3.78 months) attributed to inadequate obliteration, recanalization, or missed defects. Notable complications included intracranial hypertension and facial paralysis. CONCLUSION:Spontaneous CSF otorrhea or otorhinorrhea typically originates from the middle cranial fossa floor in adults. While multi-layered surgical repair is effective, its success can be compromised by technical factors, underscoring the necessity of meticulous preoperative assessment to reduce recurrence.

Cerebral palsy (CP) is a leading cause of childhood motor disability with a notable male predominance, suggesting that X-linked genetic factors may contribute to CP susceptibility. Although SHROOM4 has been implicated in several neurodevelopmental disorders, its role in CP remains unclear. This study aimed to investigate the contribution of SHROOM4 variants to male CP susceptibility. Whole-exome sequencing was performed in 1,010 Chinese male patients with sporadic CP and 1,014 male controls. Association analysis focused on common variants and haplotypes within SHROOM4. Rare SHROOM4 variants identified in CP patients were further validated and characterized using qPCR, immunofluorescence, western blotting and CRISPR/Cas9-mediated knockout cell lines. A common T-A-G haplotype comprising rs2873098, rs2295544 and rs2295543 in SHROOM4 was significantly associated with male CP susceptibility (OR = 6.091, Pc = 3.26E-07) and was enriched in CP patients with intrauterine growth restriction. Additionally, a rare nonsense variant, c.C2050T (p.Arg684*), was identified in a patient presenting with spastic CP and intellectual disability. Functional analyses showed that p.Arg684* and a population-derived frameshift variant (p.Glu1140fs*42) were associated with reduced SHROOM4 transcript abundance, consistent with NMD-mediated transcript reduction, while residual mutant transcripts produced detectable truncated proteins with variant-specific effects on protein stability, subcellular localization, and actin cytoskeletal organization. Our findings support SHROOM4 as an X-linked susceptibility gene associated with male CP, suggesting that both common haplotypes and rare SHROOM4 variants may contribute to the genetic susceptibility to CP. These results expand our current understanding of the genetic architecture of CP and highlight cytoskeletal regulation as a potentially relevant mechanism associated with SHROOM4 variants.

Postoperative cognitive dysfunction (POCD) is common in older surgical patients, yet its mechanistic underpinnings remain incompletely defined. Building on the premise that immune-cell metabolism shapes inflammatory signaling, we investigated whether microglia export metabolic instructions through extracellular vesicles (EVs) to remodel astrocytic function. We show that plasma EVs from POCD patients and EVs from activated microglia are enriched for ATP citrate lyase (ACLY) and are efficiently taken up by astrocytes. EV-delivered ACLY increases acetyl-CoA availability and enhances NF-κB p65 acetylation, which in turn suppresses the astrocytic glutamate transporter EAAT1 (SLC1A3). This program diminishes glutamate clearance, leading to synaptic dysfunction and cognitive impairment. In aged mice, systemic or hippocampal EV administration recapitulated microgliosis/astrogliosis, dendritic spine loss, impaired long-term potentiation, and deficits in the Morris water maze and novel object recognition. Chromatin immunoprecipitation confirmed acetyl-p65 occupancy at the SLC1A3 promoter, linking ACLY activity to direct transcriptional repression of EAAT1. Interventions that blocked EV biogenesis/uptake or reduced ACLY (genetically or pharmacologically) attenuated p65 acetylation, restored EAAT1 expression and glutamate uptake, and improved synaptic and behavioral outcomes. These findings identify a microglia-to-astrocyte ACLY-p65-EAAT1 axis that couples immunometabolism to excitatory neurotransmitter dysregulation and cognitive decline in POCD, highlighting ACLY and EV pathways as actionable therapeutic targets.

Cerebral palsy (CP) is a neurodevelopmental disorder with a significant male predisposition, yet the underlying genetic mechanisms driving this sex-specific risk remain poorly understood. Given the hemizygous state of X-linked variants in males, we hypothesized that X-chromosomal genetic variations may contribute to CP susceptibility in male patients. We performed an X-chromosome-wide association study (XWAS) using whole-exome sequencing (WES) data from 1,076 male CP patients and 1,057 healthy male controls of Chinese ancestry. Quality control, principal component analysis (PCA), and logistic regression were applied to identify risk loci for CP. Six candidate single nucleotide variants (SNVs) in UTP14A were genotyped via MassARRAY, and haplotype analysis was conducted using SHEsis. We identified a significant association between the intronic variant rs2281277 in UTP14A and CP (P = 1.38E-05, FDR q-value = 0.0465, OR = 0.644). In addition to the lead SNV rs2281277, five other SNVs in UTP14A also showed a suggestive association with CP, including rs141750783, rs61318448, rs2273021, rs2281278, and rs111975985. Haplotype analysis further revealed a risk haplotype (CGTATC) defined by these six SNVs, which was associated with a 53.1

BACKGROUND AND AIMS:USP53 variants cause low-GGT progressive familial intrahepatic cholestasis (PFIC). The mechanism is not well understood. USP53, which encodes a ubiquitin-specific protease, has been proposed to be involved in blood-bile barrier impairment. However, Usp53 knockout mice did not show blood-bile barrier impairment. The aim of this study was to investigate the molecular mechanism underlying USP53 -PFIC. APPROACH AND RESULTS:Immunohistochemistry of patient tissue, confocal immunofluorescence microscopy and surface protein biotinylation were performed to investigate the localization of proteins of interest. Small-interference RNA and CRISPR-Cas9 technology were used to downregulate or knock out genes, respectively. Site-directed mutagenesis was performed to generate gene variants for expression in cells. Co-immunoprecipitation experiments were performed to investigate (variant) protein-protein interactions. Live cell imaging and fluorescence recovery after photobleaching were performed to investigate protein dynamics. The mislocalization of the bile salt export pump (BSEP) was demonstrated in hepatocytes of a USP53-associated PFIC patient and USP53 -KO cells. Loss of USP53 caused BSEP accumulation in MYO5B-positive and RAB11A-positive recycling endosomes and impaired BSEP trafficking to the plasma membrane. USP53 colocalized with MYO5B and interacted with its IQ domain. The recurrent MYO5B-PFIC-associated p.(Arg824Cys) variant, located in the IQ domain, failed to interact with USP53. Loss of USP53 expression resulted in increased ubiquitination of MYO5B and interfered with the endosomal recruitment of MYO5B. CONCLUSIONS:Loss of USP53 interaction with MYO5B and its p.(Arg824Cys) variant impaired BSEP trafficking in USP53 -associated and MYO5B -associated low-GGT intrahepatic cholestasis. These results provide a novel mechanism that underlies USP53 -PFIC and implicates USP53 in the pathogenesis of MYO5B -PFIC.

The dynamic nature of protein conformations is central to their biological functions. Conventional structural biology techniques provide static snapshots, whereas a comprehensive understanding requires an analysis of the dynamic conformations. In this study, we develop a transient cross-linking mass spectrometry method using a photo-cross-linker DCD. This cross-linker can be transiently activated to accomplish cross-linking, and with sample freezing, transient conformations are preserved, allowing temporal control and on-demand cross-linking. Its cross-linking site covers all amino acids, exhibiting diversity and providing rich structural information. Additionally, we develop a data-processing strategy by integrating a DCD-specific reporter ion and a defined ambiguous site annotation criterion, thereby ensuring the confidence in identification and cross-link site annotation. Thus, the developed transient cross-linking mass spectrometry, leveraging the distinctive features of DCD, has enabled us to analyze protein conformations and protein complexes with high resolution, take conformational snapshots, discern the coexistence of conformational intermediates, and decipher conformational fluctuations, shedding light on how proteins conformationally respond to biological signals and engage with interacting partners. Our results highlight DCD's potential for probing protein conformational changes, facilitating the elucidation of their pivotal roles within biological systems.

Background Noninvasive prenatal diagnosis (NIPD) for autosomal recessive nonsyndromic hearing loss (ARNSHL) was rarely reported until recent years. However, previous method would be unavailable without the proband genotype or challenging genome loci(e.g. CNV, deletions, inversions, or gene recombinants). Here, We assess the performance of relative haplotype dosage analysis (RHDO) - based NIPD for identifying fetal genotyping of pregnancies at risk of ARNSHL. Methods Fifty couples carrying pathogenic variants associated with ARNSHL in either GJB2 or SLC26A4 were recruited. The fetal cell-free DNA (cfDNA) of 49 families that met the quality control standard was genotyped using RHDO-based targeted linked-read sequencing. Fetal amniocyte samples were genotyped by invasive prenatal diagnosis (IPD) to evaluate the performance of NIPD. Results The 49 fetal genotypes were identified as 14 normal homozygotes, 17 heterozygotes, and 18 affected homozygotes or compound heterozygotes by NIPD, which were all concordant with those diagnosed by IPD. The concordance rate was 100% for the GJB2 (31/31) as well as SLC26A4 (18/18) genotyping. The sensitivity and specificity were both 100%. Conclusions Sufficient specific informative SNPs for haplotyping, the fetal cfDNA concentration as well as the sequencing depth are prerequisites for the RHDO-based NIPD, which has the merits of being proband-free and qualified for copy number variation analysis, thus it is suitable for universal application. It also has clinical potential as an alternative to traditional IPD for ARNSHL.

Noninvasive prenatal diagnosis (NIPD) for autosomal recessive nonsyndromic hearing loss (ARNSHL) has been rarely reported until recent years. Additionally, the existing method can not be used for challenging genome loci (eg, copy number variations, deletions, inversions, or gene recombinants) or on families without proband genotype. This study assessed the performance of relative haplotype dosage analysis (RHDO)-based NIPD for identifying fetal genotyping in pregnancies at risk of ARNSHL. Fifty couples carrying pathogenic variants associated with ARNSHL in either GJB2 or SLC26A4 were recruited. The RHDO-based targeted linked- read sequencing combined with whole gene coverage probes was used to genotype the fetal cell-free DNA of 49 families who met the quality control standard. Fetal amniocyte samples were genotyped using invasive prenatal diagnosis (IPD) to assess the performance of NIPD. The NIPD results showed 100% (49/49) concordance with those obtained through IPD. Two families with copy number variation and recombination were also successfully identified. fi ed. Sufficient fi cient specific fi c informative single-nucleotide polymorphisms for haplotyping, as well as the fetal cell-free DNA concentration and sequencing depth, are prerequisites for RHDObased NIPD. This method has the merits of covering the entire genes of GJB2 and SLC26A4, qualifying for copy number variation and recombination analysis with remarkable sensitivity and specificity. fi city. Therefore, it has clinical potential as an alternative to traditional IPD for ARNSHL. (J Mol Diagn 2024, 26: 638-651; https:// doi.org/10.1016/j.jmoldx.2024.04.002)

Perinatal white matter injury (PWMI) can lead to permanent neurological damage in preterm infants and bring a huge economic burden to their families and society. Liver X receptors (LXRs) are transcription factors that have been confirmed to mediate the myelination process under physiological conditions and are involved in regulating neurogenesis in adult animal models of acute and chronic cerebral ischemia. However, the role of LXRs in PWMI induced by both ischemic and hypoxic stimulation in the immature brain has not been reported. Herein, we investigated the role of LXRs in a neonatal rat model of white matter loss after hypoxia-ischemia (HI) injury through intraperitoneal injection of the LXR agonist T0901317 (T09) 1 day before and 15 min postinjury. The in vivo data showed that T09 treatment significantly facilitated myelination and ameliorated neurological behavior after PWMI. Moreover, T09 enhanced the proliferation of oligodendrocyte lineage cells and reduced microgliosis and astrogliosis in the microenvironment for oligodendrocytes (OLs), maintaining a healthy microenvironment for myelinating OLs. In vitro data suggested that the expression of the myelin-related genes Plp and Cnpase was increased in OLN-93 cells after T09 intervention compared with OLN-93 cells injured by oxygen and glucose deprivation (OGD). In primary mixed astrocytes/microglia cells, T09 also reduced the expression of Il6, Cox2, Tnfa and Il10 that was induced by OGD. Mechanistically, the mRNA expression level and the protein level of ATP binding cassette subfamily A member 1 (Abca1) decreased after HI injury, and the protective effect of T09 might be related to the activation of the LXR beta-ABCA1 signaling pathway. Our study revealed the protective role of LXRs in myelination and white matter homeostasis, providing a potential therapeutic option for PWMI.

Waardenburg syndrome(WS)is an autosomal-dominant neurocristopathy characterized by sensorineural hearing loss(SNHL)and pigmentary abnormalities of the iris,hair,and skin.It is responsible for approximately 3％of congenital hearing loss[1].WS is thought to be fully penetrant when considering at least one clinical feature,but the penetrance of each feature is not complete[2].

Pathogenic variants in MYO15A are known to cause autosomal recessive nonsyndromic hearing loss (ARNSHL), DFNB3. We have previously reported on one ARNSHL family including two affected siblings and identified MYO15A c.5964+3G > A and c.8375 T > C (p.Val2792Ala) as the possible deafness-causing variants. Eight year follow up identified one new affected individual in this family, who also showed congenital, severe to profound sensorineural hearing loss. By whole exome sequencing, we identified a new splice-site variant c.5531+1G > C (maternal allele), in a compound heterozygote with previously identified missense variant c.8375 T > C (p.Val2792Ala) (paternal allele) in MYO15A as the disease-causing variants. The new affected individual underwent unilateral cochlear implantation at the age of 1 year, and 5 year follow-up showed satisfactory speech and language outcomes. Our results further indicate that MYO15A -associated hearing loss is good candidates for cochlear implantation, which is in accordance with previous report. In light of our findings and review of the literatures, 58 splice-site variants in MYO15A are correlated with a severe deafness phenotype, composed of 46 canonical splice-site variants and 12 non-canonical splice-site variants.

Abstract Cerebral palsy (CP) is the most common physical disability in childhood that results from the interaction of environmental and genetic factors. Yet in many patients, the etiology remains unknown. We identified significant association at rs3131787 within the human leukocyte antigen (HLA) region using two-stage association study between 1,090 CP cases and 1,100 controls. Fine mapping of the HLA region indicated that the carrier frequency of HLA-B*13:02 was significantly higher in CP, particularly in CP without preterm birth, low birth weight, birth asphyxia or periventricular leukomalacia (PVL). DRB1*07:01/DQA1*02:01 was also significantly enriched in CP and more specifically in dyskinetic type. Additionally, significant enrichment of carrier frequency was detected for HLA-A*32:01 in CP with either preterm birth or low birth weight and for HLA-B*27:05 in CP with birth asphyxia. These data suggest that immune dysregulation resulting from immunogenetic variants or environmental exposures may underlie the pathogenesis of CP.

Hereditary hearing loss is genetically heterogeneous, with diverse clinical manifestations. Here we performed targeted genome sequencing of 227 hearing loss related genes in 1027 patients with bilateral hearing loss and 520 healthy volunteers with normal hearing to comprehensively identify the molecular etiology of hereditary hearing loss in a large cohort from China. We obtained a diagnostic rate of 57.25% (588/1027) for the patients, while 4.67% (48/1027) of the patients were identified with uncertain diagnoses. Of the implicated 35 hearing loss genes, three common genes, including SLC26A4 (278/588), GJB2 (207/588), MT-RNR1 (19/588), accounted for 85.54% (503/588) of the diagnosed cases, while 32 uncommon hearing loss genes, including MYO15A , MITF , OTOF , POU3F4 , PTPN11 , etc. accounted for the remaining diagnostic rate of 14.46% (85/588). Apart from Pendred syndrome, other eight types of syndromic hearing loss were also identified. Of the 64 uncertain significant variants and 244 pathogenic/likely pathogenic variants identified in the patients, 129 novel variants were also detected. Thus, the molecular etiology presented with high heterogeneity with the leading causes to be SLC26A4 and GJB2 genes in the Chinese hearing loss population. It’s urgent to develop a database of the ethnicity-matched healthy population as well as to perform functional studies for further classification of uncertain significant variants.

Abstract Background Surgical intervention can effectively treat venous pulsatile tinnitus. Aim/objectives To assess the effectiveness of treating of venous pulsatile tinnitus (VPT) by compression reconstruction of sigmoid sinus (SSCR) under local anesthesia. Material and methods This study retrospectively reviewed 41 patients with VPT in our otolaryngology department between September 2009 and February 2019. Under local anesthesia, all patients were received SSCR. Pre- and postoperative Tinnitus Handicap Inventory (THI) degree and scores were used to evaluate the efficacy of SSCR for VPT. Results Of the 41 patients, 36 patients were followed up from 9 months to 8 years and 5 patients were lost to follow-up and were excluded from the data analysis. SSCR was clinically effective in 86% of patients with complete disappearance in 18 patients (50%), partial remission in 10 patients (28%), slight alleviation in 3 patients (8%), and no change in 5 patients (14%). The pre- and postoperative THI degree and scores were significantly different (p < .001 and p = .002, respectively). Conclusions and significance SSCR under local anesthesia is effective for treating patients with VPT. It is critical to perform a rigorous pre-operative clinical and radiological evaluation to reduce intra- and postoperative complications.

Cochlear implantation (CI) is a safe and beneficial surgery for children with congenital inner ear malformations, with the exception of cochlear nerve aplasia. The combination of microtia with middle and inner ear abnormalities is extremely uncommon and sufficiently severe to make a surgical approach to the cochlea difficult. We report herein the case of a 2-year-old girl who presented with profound bilateral sensorineural hearing loss, congenital aural atresia, microtia, and inner ear malformations. High-resolution computed tomography revealed poor development of the bilateral middle ear spaces, absence of the incus and stapes, aberrant courses of facial nerves, aplastic lateral semicircular canals, and covered round windows. With intraoperative imaging assistance, sequential bilateral CI was performed using a transmastoid approach with no complication. We propose that CI is feasible in patients with severe external and middle ear malformations. However, major malformations increase the risk of complications. As the facial nerve and cochlea are difficult to locate due to the lack of important anatomical landmarks, detailed planning and adequate preparation, including review of the preoperative imaging data, and the use of facial nerve monitoring and intraoperative imaging are very important. In addition, experienced surgeons should perform CI to ensure the success of the operation.

Waardenburg syndrome (WS) is a phenotypically and genetically heterogeneous disorder characterised by hearing loss and pigmentary abnormalities. We clarified the clinical and genetic features in 90 Chinese WS probands. Disease-causing variants were detected in 55 probands, for a molecular diagnosis rate of 61%, including cases related to PAX3 (14.4%), MITF (24.4%), and SOX10 (22.2%). Altogether, 48 variants were identified, including 44 single-nucleotide variants and 4 copy number variants. By parental genotyping, de novo variants were observed in 60% of probands and 15.4% of the de novo variation was associated with mosaicism. Statistical analyses revealed that brown freckles on the skin were more frequently seen in probands with MITF variants; patchy depigmented skin, asymmetric hearing loss, and white forelocks occurred more often in cases with PAX3 variants; and congenital inner ear malformations were more common and cochlear hypoplasia III was exclusively observed in those with SOX10 variants. In addition, we found that ranges of W-index values overlapped between WS probands with different genetic variants, and the use of the W-index as a tool for assessing dystopia canthorum may be problematic in Chinese. Herein, we report the spectrum of a cohort of WS probands and elucidate the relationship between genotype and phenotype.