Objective:The aim of this study is to analyze the clinical characteristics and genetic variants of a late-onset auditory neuropathy pedigree caused by maternally inherited- mitochondrial mutation.Methods:A male proband who presented with bilateral sensorineural hearing loss at Henan Children′s Hospital in September 2023 was chosen, along with his family members (4 generations, 20 individuals) as the study subjects. Data from this pedigree were collected, organized, and analyzed for clinical genetic characteristics. Medical histories were obtained from family members, pedigree charts were drawn, audiological, imaging, and physical examinations were conducted. Pathogenic genes and mutations were screened using high-throughput sequencing. Sanger sequencing was employed for variant confirmation and segregation validation in the family.Results:In this family, a total of 12 members (10 members collected) had sensorineural hearing loss, characterized by late-onset hearing impairment with an onset age ranging from 9 to 30 years. The patients exhibited poor speech recognition rates, and audiometric examinations are consistent with auditory neuropathy. There was no history of ototoxic drug use. High-throughput sequencing identified the variant NC_012920.1:m.7471dup in the mitochondrial MT-TS1 gene as the pathogenic variant. Sanger sequencing results confirmed that the pathogenic gene mutation site perfectly co-segregated with the auditory neuropathy phenotype in this family. According to the classification criteria and guidelines for genetic variations by the American College of Medical Genetics and Genomics, the variant was classified as a pathogenic mutation. Conclusion:The mitochondrial MT-TS1 gene mutation m.7471dup is considered to be the pathogenic cause in this late-onset auditory neuropathy pedigree.

Objective To analyze signals of adverse events(AEs)related to macrolide antibiotic in children from the US FDA Adverse Event Reporting System(FAERS),and provide a reference for clinical safe medication in children.Methods The AEs reports of children(less than 18 years old)were extracted from FAERS during January 2004 to March 2024,and the reports of odds ratio(ROR)and information component of Bayesian confidence propagation neural network(IC)were used to analyze the signals of macrolide antibiotics in children.Results There were 2 133 648 reports of children from FAERS,including azithromycin(7 589 cases),clarithromycin(3 673 cases),and erythromycin(820 cases).The significant signals of system organ class(SOC)were 16 in total,such as skin disorders,ear and labyrinth disorders,gastrointestinal disorders,nervous system,immune system disorders,etc.A proportional imbalance analysis was performed on macrolide antibiotics(azithromycin and clarithromycin),and obtained relevant positive signals,including azithromycin(232)and clarithromycin(194).The most frequently reported AEs included rash,vomiting,diarrhea,hypersensitivity and so on.The strongest signals of azithromycin were Stevens-Johnson syndrome(SJS),hypersensitivity reactions,drug-induced liver injury,etc.,while the strongest signals of clarithromycin were dysgeusia,lip swelling,toxic epidermal necrolysis(TEN),nightmare,psychotic disorder,etc.17 unlisted signals of azithromycin and 21 of clarithromycin were found.The same PT of azithromycin and clarithromycin had significant difference,such as dyspnea,hypersensitivity,rash,SJS,vomiting,and more.Conclusions To ensure the safety of children's treatment,children using macrolide should pay attention to skin,gastrointestinal,ear and other toxicity and hypersensitivity reactions,especially serious skin reactions,unlisted suspicious AEs and psychiatric disorders.

Objective To analyze signals of adverse events(AEs)related to macrolide antibiotic in children from the US FDA Adverse Event Reporting System(FAERS),and provide a reference for clinical safe medication in children.Methods The AEs reports of children(less than 18 years old)were extracted from FAERS during January 2004 to March 2024,and the reports of odds ratio(ROR)and information component of Bayesian confidence propagation neural network(IC)were used to analyze the signals of macrolide antibiotics in children.Results There were 2 133 648 reports of children from FAERS,including azithromycin(7 589 cases),clarithromycin(3 673 cases),and erythromycin(820 cases).The significant signals of system organ class(SOC)were 16 in total,such as skin disorders,ear and labyrinth disorders,gastrointestinal disorders,nervous system,immune system disorders,etc.A proportional imbalance analysis was performed on macrolide antibiotics(azithromycin and clarithromycin),and obtained relevant positive signals,including azithromycin(232)and clarithromycin(194).The most frequently reported AEs included rash,vomiting,diarrhea,hypersensitivity and so on.The strongest signals of azithromycin were Stevens-Johnson syndrome(SJS),hypersensitivity reactions,drug-induced liver injury,etc.,while the strongest signals of clarithromycin were dysgeusia,lip swelling,toxic epidermal necrolysis(TEN),nightmare,psychotic disorder,etc.17 unlisted signals of azithromycin and 21 of clarithromycin were found.The same PT of azithromycin and clarithromycin had significant difference,such as dyspnea,hypersensitivity,rash,SJS,vomiting,and more.Conclusions To ensure the safety of children's treatment,children using macrolide should pay attention to skin,gastrointestinal,ear and other toxicity and hypersensitivity reactions,especially serious skin reactions,unlisted suspicious AEs and psychiatric disorders.

Objective:The aim of this study is to analyze the clinical characteristics and genetic variants of a late-onset auditory neuropathy pedigree caused by maternally inherited- mitochondrial mutation.Methods:A male proband who presented with bilateral sensorineural hearing loss at Henan Children′s Hospital in September 2023 was chosen, along with his family members (4 generations, 20 individuals) as the study subjects. Data from this pedigree were collected, organized, and analyzed for clinical genetic characteristics. Medical histories were obtained from family members, pedigree charts were drawn, audiological, imaging, and physical examinations were conducted. Pathogenic genes and mutations were screened using high-throughput sequencing. Sanger sequencing was employed for variant confirmation and segregation validation in the family.Results:In this family, a total of 12 members (10 members collected) had sensorineural hearing loss, characterized by late-onset hearing impairment with an onset age ranging from 9 to 30 years. The patients exhibited poor speech recognition rates, and audiometric examinations are consistent with auditory neuropathy. There was no history of ototoxic drug use. High-throughput sequencing identified the variant NC_012920.1:m.7471dup in the mitochondrial MT-TS1 gene as the pathogenic variant. Sanger sequencing results confirmed that the pathogenic gene mutation site perfectly co-segregated with the auditory neuropathy phenotype in this family. According to the classification criteria and guidelines for genetic variations by the American College of Medical Genetics and Genomics, the variant was classified as a pathogenic mutation. Conclusion:The mitochondrial MT-TS1 gene mutation m.7471dup is considered to be the pathogenic cause in this late-onset auditory neuropathy pedigree.

Objective To investigate the characteristics of pathogenic bacteria in cancer patients with severe infection after radiotherapy and chemotherapy and the application value of metagenomic next-generation sequencing(NGS)in the detection of pathogenic bacteria.Methods A total of 112 patients with severe infection after malignant tumor radiation and chemotherapy admitted to Department of Critical Care Medicine of Wuwei Tumor Hospital from October 2019 to August 2023 were selected as the research objects.A total of 150 specimens from suspected infection sites were collected and tested by traditional etiology and NGS.The characteristics of pathogenic bacteria in severe infection of cancer patients after chemoradiotherapy and the application value of NGS in pathogen detection were analyzed.Results Among 150 samples of 112 patients with severe infection after radiotherapy and chemotherapy,the highest proportion of respiratory system infection was 51.79%(58 cases),followed by 25.89%(29 cases)of bloodstream infection,the lowest central nervous system infection rate was 1.79%(2 cases).Gram-negative bacteria were the most common pathogenic bacteria[NGS 35.33%(53 cases),traditional pathogen detection 23.33%(35 cases)],followed by Gram-positive bacteria[NGS 20.67%(31 cases),traditional pathogen detection 12.00%(18 cases)],and multi-drug resistant bacteria infection rate was more than 80.00%,multi-drug resistant fungal infection rate also reached 28.57%.In the patients with severe infection after radiotherapy and chemotherapy,the positive rate of pathogenic bacteria of NGS was significantly higher than that of traditional pathogen detection[87.33%(131/150)vs.42.67%(64/150),P＜0.01],and the positive detection rate of Gram-negative bacteria,Gram-positive bacteria,fungi and other pathogens was significantly higher than that of traditional pathogen detection[Gram-negative bacteria:35.33%(53/150)vs.23.33%(35/150),Gram-positive bacteria:20.67%(31/150)vs.12.00%(18/150),fungi:22.67%(34/150)vs.6.67%(10/150),others:8.67%(13/150)vs.0.67%(1/150),both P＜0.05].Conclusions The detection rate of pathogenic bacteria of NGS in patients with severe infection after radiotherapy and chemotherapy was significantly higher than that of traditional pathogen detection.The distribution and drug resistance of pathogenic bacteria in severe infection after radiotherapy and chemotherapy are unique,and strengthening the detection of pathogenic bacteria is helpful to improve the curative effect.

Objective:To investigate the molecular etiology of Perrault syndrome by analyzing the clinical phenotype and pathogenic gene variants of 2 male patients with bilateral severe sensorineural deafness.Methods:Two male patients with Perrault syndrome characterized by severe sensonrineual deafness adimitted to the First Affiliated Hospital of Zhengzhou University between February 2021 and March 2022 were selected, and the clinical phenotype and pathogenic gene variants of them and their family members were summarized. The whole exome sequencing technology was used to screen the pathogenic variants of the probands, and the candidate variants were determined by combining with clinical phenotype. The probands and their family members were verified by the Sanger sequencing method.Results:The whole exome sequencing results showed that the proband of family 1 had a compound heterozygous variants of the LARS2 (NM_015340.4) gene c.1565C>A (p.Thr522Asn) and c.1079T>C (p.Ile360Thr). The reported pathogenic variant c.1565C>A came from the mother, and the novel variant c.1079T>C came from the father. The second proband harbored compound heterozygous variants of HARS2 gene (NM_012208.4) c.1273C>T (p.Arg425Trp) and c.1403G>C (p.Gly468Ala), with the former from the proband′s mother, the latter from the father. The c.1273C>T was novel and c.1403G>C was the reported pathogenic variant. All above variants were respectively classified as pathogenic, uncertain significance, uncertain significance and likely pathogenic based on the ACMG guidelines. Conclusion:This study expands the mutational spectrum of LARS2 and HARS2 genes, which highlights that genetic testing plays an important role in the early diagnosis of syndromic deafness.

Objective:To screen the causative genes of five families with branchio-oto-renal syndrome (BORS) or branchio-oto syndrome(BOS) and to analyze the phenotypic characteristics and clinical management strategies of patients.Methods:Five families with BORS/BOR from December 2018 to September 2021 were recruited, information of patients, including family history and medical history, was collected, and genealogies were drawn. The examinations concerning audiology, nephrology, and radiology were performed on the affected individuals. Peripheral blood was obtained for DNA extraction, then next-generation sequencing technology was used to screen candidate variants associated with BORS/BOS. Based on patient′s clinical results, the appropriate interventions were recommended and implemented.Results:Eight individuals were diagnosed with BOS or BORS. Of the eight patients, all had hearing loss, preauricular pits and ear malformations, and only four presented with branchial cleft fistulae or cysts. Except for two patients(5-I-2, 5-II-2) who did not undergo renal examination, the remaining six lacked renal abnormalities. Genetic analysis identified four likely pathogenic or pathogenic EYA1 variants (c.1715G>T, c.1140+1G>A, c.639G>C, c.1475+1G>C; NM_000503.6), and c.1715G>T was first reported in this study. Middle ear ossicular reconstruction was performed in 1-II-2,2-I-2 and 3-II-2, but did not yield the expected results; then hearing aids and cochlear implantation were recommended and achieved satisfactory results. Conclusions:Next-generation sequencing technology facilitates the diagnosis and genetic counseling of BORS/BOS. Hearing loss, preauricular pits, ear malformations and branchial cleft fistulae or cysts are the most common manifestations of patients in this study. Middle ear surgeries for improving hearing loss may have some limitations in BORS/BOS patients, and hearing aids and cochlear implantation can contribute to hearing gains.

Objective:To explore the pathogenic variants of a family with syndromic deafness by high-throughput sequencing.Methods:The family was from Puyang City, Henan Province, and had four members, including two with syndromic deafness. The proband and his sister had congenital deafness, and their parents had normal phenotypes. The clinical phenotype of the family was characterized using clinical examinations and pedigree analysis. The clinical examinations included imaging examination, audiometry (pure tone audiometry, acoustic immittance, brainstem auditory evoked potential, and otoacoustic emission), vestibular function test, and ophthalmic examination (visual acuity test, visual field test, fundus examination, visual evoked potential, and electroretinogram). Target exome sequencing of 129 known deafness genes and bioinformatics analysis were used to screen suspected pathogenic variants. Sanger sequencing and minigene assay were used to verify and functionally investigate the mutation detected, respectively. According to the standards and guidelines for interpreting genetic variants proposed by the American College of Medical Genetics and Genomics, the variants c.6049G>A and c.8699A>G were classified as pathogenic/likely pathogenic, and the variant c.9856C>G was classified as variants of uncertain significance.Results:The probands and his sister had severe sensorineural hearing loss with decreased binocular vision, night blindness, decreased peripheral visual field sensitivity and partial visual field defect, and normal vestibular function. Both of them had three CDH23 mutations, including CDH23 (NM_022124.5) c.6049G>A (p.Gly2017Ser),c.9856C>G (p.His3286Asp), and c.8699A>G (p. Asp2900Gly), The first two were inherited from the father, and the last one was from the mother. The missense variants c.9856C>G and c.8699A>G were not included in the gnomad database. The missense mutation c.6049G>A was located in the last position of exon 46 and was predicted to affect splicing by bioinformatics software. The minigene experiment showed that the mutation cause exon skipping of exon 46, resulting in an abnormal protein. Conclusions:Compound heterozygous variations of the CDH23 are the leading cause of USH1D in the family. This study confirms that the compound heterozygosity of splicing and missense variants of the CDH23 gene could lead to USH1D.

Objective: To determine the extent of transfection and expression of adeno-associated virus (AAV) serotype 9 (AAV9) in the cochleae of mice at different ages. Methods: AAV9-green fluorescent protein (GFP) was inoculated into the cochlea of mice via the round window membrane (RWM) or through cochleostomy at different ages. Four groups were divided according to ages and injection sites: P1SM group, AAV9-GFP was delivered to the scala media by cochleostomy at postnatal day 1; P1RW group, AAV9-GFP was delivered to the scala tympani via RWM at postnatal day 1; P9RW group: AAV9-GFP was injected through RWM at postnatal day 9; and P30RW group, adult mice (P30) were injected through RWM. GFP expression in cochlear whole mount was analyzed and auditory brainstem response (ABR) tests were conducted one month after virus injection (for each animal, only left cochlea was injected and the right side was used as a control). GraphPad Prism 5 statistical software was used for data analysis. Results: All of inner hair cells (IHCs) and most of outer hair cells (OHCs) were transfected via two approaches at P1 injection. There was no significant difference in ABR threshold between injected ears and untreated ears (P>0.05). All of the IHCs and parts of OHCs (69% in apical turn) were transfected via RWM at P9. The strongest GFP expression was observed near the apical turn. Cochlear inoculation via RWM at P30 led to transgene expression in only IHCs. The ABR threshold of injected ears in P9RW group and P30RW group was significantly higher than that of contralateral ears (P<0.01). Conclusions AAV9 can be highly expressed in the inner and outer hair cells of the cochlea and hearing sensitivity can be preserved if virus injections are performed in neonatal mice. After AAV9 is transfected into the inner ear of adult mice, it is only expressed in the inner hair cells, which leads to the increase of the ABR response threshold of mice. Transfection efficiency is significant higher in neonate mice than in P9 and adult mice.

Objective Recent studies have indicated that early brain injury is the leading cause of death in patients with subarachnoid hemorrhage ( SAH) .Our study investigated the role of aminoguanidine ( AG) in early brain injury after SAH . Methods Sixty-eight male SD rats were equally randomized into four groups of equal number :control, sham, SAH, and AG.The animals in the sham group were injected with isotonic saline solution , while those of the latter two groups with femoral artery blood ( FAB) and FAB+AG, respectively, into the pre-chiasmatic cistern to induce SAH. At 24 hours after modeling , all the rats were killed for HE staining , obtainment of behavioral neurological assessment ( BNA ) scores by Garcia, measurement of the apoptosis of neurons by TUNEL , and de-termination of the expressions of the iNOS and NSE proteins by West-ern blot. Results The results of HE staining showed the presence of more red blood cells in the subarachnoid cavity of the rats in the SAH group, with a significantly decreased BNA score ( 14.47 ± 0.62) as compared with those in the control (17.94 ±0.24), sham (17.59 ±0.51), and AG group (15.71 ±0.47) (P<0.05). The rate of positive cells was remarkably higher in the SAH group ([42.38 ±2.38]%) than in the control ([6.35 ±0.94]%), sham ([6.85 ±0.69]%), and AG group ([30.48 ±2.89]%) ( P<0.01), with significant differences among the latter three groups (P<0.05).The expressions of iNOS and NSE were markedly higher in the SAH group ([3.86 ±0.07] and [1.59 ±0.06]) than in the control (0 and[0.35 ±0.09]), sham ([2.96 ±0.34] and [0.38 ±0.08]), and AG group ([3.41 ±0.04] and [0.70 ±0.12]) ( P<0.05).Both the expression levels of iNOS and NSE were positively correlated with the rate of positive cells (r=0 .879 and 0.935, P<0.01). Conclusion AG can alleviate early brain injury after SAH in SD rats by improving the neuro-ethologic function , suppressing the apoptosis of neurons , and reducing the expressions of iNOS and NSE .