DNA methylation regulates the secondary metabolism of saponins to improve the adaptability of Eleutherococcus senticosus during drought stress.

Plant growth and development can be significantly impacted by drought stress. Plants will adjust the synthesis and accumulation of secondary metabolites to improve survival in times of water constraint. Simultaneously, drought stress can lead to modifications in the DNA methylation status of plants, and these modifications can directly impact gene expression and product synthesis by changing the DNA methylation status of functional genes involved in secondary metabolite synthesis. However, further research is needed to fully understand the extent to which DNA methylation modifies the content of secondary metabolites to mediate plants' responses to drought stress, as well as the underlying mechanisms involved. Our study found that in Eleutherococcus senticosus (E. senticosus), moderate water deprivation significantly decreased DNA methylation levels throughout the genome and at the promoters of EsFPS, EsSS, and EsSE. Transcription factors like EsMYB-r1, previously inhibited by DNA methylation, can re-bind to the EsFPS promotor region following DNA demethylation. This process promotes gene expression and, ultimately, saponin synthesis and accumulation. The increased saponin levels in E. senticosus acted as antioxidants, enhancing the plant's adaptability to drought stress.

Identification and validation of chemokine system-related genes in idiopathic pulmonary fibrosis.

Background: Idiopathic pulmonary fibrosis (IPF) is a chronic progressive interstitial lung disease with limited therapeutic options. Recent studies have demonstrated that chemokines play a vital role in IPF pathogenesis. In the present study, we explored whether the gene signature associated with chemokines could be used as a reliable biological marker for patients with IPF. Methods: Chemokine-related differentially expressed genes (CR-DEGs) in IPF and control lung tissue samples were identified using data from the Gene Expression Omnibus database. A chemokine-related signature of the diagnostic model was established using the LASSO-Cox regression. In addition, unsupervised cluster analysis was conducted using consensus-clustering algorithms. The CIBERSORT algorithm was used to calculate immune cell infiltration across patient subgroups. Finally, we established a mouse model of bleomycin-induced pulmonary fibrosis and a model of fibroblasts treated with TGFß1. Expression levels of chemokine-related signature genes were determined using real-time quantitative polymerase chain reaction (RT-qPCR). Results: We established a chemokine-related eleven-gene signature of a diagnostic model consisting of CXCL2, CCRL2, ARRB1, XCL1, GRK5, PPBP, CCL19, CCL13, CCL11, CXCL6, and CXCL13, which could easily distinguish between IPF patients and controls. Additionally, we identified two subtypes of IPF samples based on chemokine-related gene expression. Pulmonary function parameters and stromal scores were significantly higher in subtype 1 than in subtype 2. Several immune cell types, especially plasma cells and macrophages, differ significantly between the two subtypes. RT-qPCR results showed that the expression levels of Cxcl2 and Ccl2 increased considerably in bleomycin-induced mice. Meanwhile, Arrb1, Ccrl2, Grk5, and Ppbp expression was significantly reduced. Furthermore, multiple chemokine-related genes were altered in TGFß1 or TNFα-induced fibroblast cells. Conclusions: A novel chemokine-related eleven-signature of diagnostic model was developed. These genes are potential biomarkers of IPF and may play essential roles in its pathogenesis.

Evolution of elemental nitrogen involved in the carbonization mechanism and product features from wet biowaste.

Hydrothermal carbonization (HTC) represents elegant thermochemical conversion technology suitable for energy and resource recovery from wet biowaste, while the elemental nitrogen is bound to affect the HTC process and the properties of the products. In this review, the nitrogen fate during HTC of typical N-containing-biowaste were presented. The relationship between critical factors involved in HTC like N/O, N/C, N/H, solid ratio, initial N in feedstock, hydrothermal temperature and residence time and N content in hydrochar were systematic analyzed. The distribution and conversion of N species along with hydrothermal severity in hydrochar and liquid phase was discussed. Additionally, the chemical forms of nitrogen in hydrochar were elaborated coupled with the role of N element during hydrochar formation mechanism and the morphology features. Finally, the future challenges of nitrogen in biowaste involved in HTC about the formation and regulation mechanism of hydrochar were given, and perspectives of more accurate regulation of the physicochemical characteristics of hydrochar from biowaste based on the N evolution is expected.

Genome-wide identification of MBD gene family members in Eleutherococcus senticosus with their expression motifs under drought stress and DNA demethylation.

BACKGROUND: Methyl-binding domain (MBD) is a class of methyl-CpG-binding domain proteins that affects the regulation of gene expression through epigenetic modifications. MBD genes are not only inseparable from DNA methylation but have also been identified and validated in various plants. Although MBD is involved in a group of physiological processes and stress regulation in these plants, MBD genes in Eleutherococcus senticosus remain largely unknown. RESULTS: Twenty EsMBD genes were identified in E. senticosus. Among the 24 chromosomes of E. senticosus, EsMBD genes were unevenly distributed on 12 chromosomes, and only one tandem repeat gene existed. Collinearity analysis showed that the fragment duplication was the main motif for EsMBD gene expansion. As the species of Araliaceae evolved, MBD genes also evolved and gradually exhibited different functional differentiation. Furthermore, cis-acting element analysis showed that there were numerous cis-acting elements in the EsMBD promoter region, among which light response elements and anaerobic induction elements were dominant. The expression motif analysis revealed that 60% of the EsMBDs were up-regulated in the 30% water content group. CONCLUSIONS: By comparing the transcriptome data of different saponin contents of E. senticosus and integrating them with the outcomes of molecular docking analysis, we hypothesized that EsMBD2 and EsMBD5 jointly affect the secondary metabolic processes of E. senticosus saponins by binding to methylated CpG under conditions of drought stress. The results of this study laid the foundation for subsequent research on the E. senticosus and MBD genes.

Hearing thresholds elevation and potential association with emotional problems among 1,914 children in Beijing, China.

Objectives: School-aged children may experience hearing loss and emotional problems. Previous studies have shown a bidirectional relationship between hearing loss and emotional problems in the elderly population, and we aimed to analyze the association between hearing thresholds and emotional problems in school-aged children. Methods: Based on the Beijing Child Growth and Health Cohort (PROC) study, the hearing screenings were conducted in November 2019 using pure tone audiometry. A total of 1,877 parents completed the Strengths and Difficulties Questionnaire (SDQ) to assess children's emotional and behavioral status. We used generalized linear regression analysis to assess the potential association of emotional problems with hearing thresholds, based on multiple imputed datasets with a sample size of 1,914. Results: The overall pass rate of hearing screening was 91.5%. The abnormal rate of SDQ total difficulties was 55.8%. Emotional symptoms were positively associated with left ear average hearing thresholds (ß = 0.24, 95%CI: 0.08-0.40), and right ear average hearing thresholds (ß = 0.18, 95%CI: 0.04-0.32). Conduct problems, hyperactivity/inattention, peer problems, and prosocial behaviors had no association with the pass rate of the hearing screening. Regarding emotional symptoms, boys with many fears and who are easily scared coincided with increased right ear average hearing thresholds (ß = 0.67, 95%CI: 0.01-1.33). Girls having many worries, frequently feeling unhappy and downhearted were positively associated with left and right ear average hearing thresholds, respectively (ß = 0.96, 95%CI: 0.20-1.73; ß = 0.72, 95%CI: 0.07-1.37). Conclusions: The co-occurrence of hearing problems and emotional problems of children aged 6-8 in Beijing attracts attention. It is important to address undiscovered hearing loss and emotional problems from the perspective of comorbidity driving factors.

Acoustic Change Complex Evoked by Horizontal Sound Location Change in Young Adults With Normal Hearing.

Acoustic change complex (ACC) is a cortical auditory-evoked potential induced by a change of continuous sound stimulation. This study aimed to explore: (1) whether the change of horizontal sound location can elicit ACC; (2) the relationship between the change of sound location and the amplitude or latency of ACC; (3) the relationship between the behavioral measure of localization, minimum audible angle (MAA), and ACC. A total of 36 normal-hearing adults participated in this study. A 180° horizontal arc-shaped bracket with a 1.2 m radius was set in a sound field where participants sat at the center. MAA was measured in a two-alternative forced-choice setting. The objective electroencephalography recording of ACC was conducted with the location changed at four sets of positions, ±45°, ±15°, ±5°, and ±2°. The test stimulus was a 125-6,000 Hz broadband noise of 1 s at 60 ± 2 dB SPL with a 2 s interval. The N1'-P2' amplitudes, N1' latencies, and P2' latencies of ACC under four positions were evaluated. The influence of electrode sites and the direction of sound position change on ACC waveform was analyzed with analysis of variance. Results suggested that (1) ACC can be elicited successfully by changing the horizontal sound location position. The elicitation rate of ACC increased with the increase of location change. (2) N1'-P2' amplitude increased and N1' and P2' latencies decreased as the change of sound location increased. The effects of test angles on N1'-P2' amplitude [F(1.91,238.1) = 97.172, p < 0.001], N1' latency [F(1.78,221.90) = 96.96, p < 0.001], and P2' latency [F(1.87,233.11) = 79.97, p < 0.001] showed a statistical significance. (3) The direction of sound location change had no significant effect on any of the ACC peak amplitudes or latencies. (4) Sound location discrimination threshold by the ACC test (97.0% elicitation rate at ±5°) was higher than MAA threshold (2.08 ± 0.5°). The current study results show that though the ACC thresholds are higher than the behavioral thresholds on MAA task, ACC can be used as an objective method to evaluate sound localization ability. This article discusses the implications of this research for clinical practice and evaluation of localization skills, especially for children.

Integrative Analysis of Metabolome and Transcriptome Reveals the Mechanism of Flavonoid Biosynthesis in Lithocarpus polystachyus Rehd.

Lithocarpus polystachyus Rehd has received great attention because of its pharmacological activities, such as inhibiting oxidation and lowering blood glucose and blood pressure, and flavonoids are one of its main pharmacodynamic components. It is important to understand the mechanisms of the flavonoid biosynthetic pathway of L. polystachyus, but the regulation of flavonoid biosynthesis is still unclear. In this study, differentially expressed genes and differentially accumulated metabolites in L. polystachyus were studied by integrating transcriptomics and metabolomics technologies. We confirmed the key genes involved in the flavonoid biosynthesis of L. polystachyus, including LpPAL3, LpCHS1, LpCHS2, LpCHI2, and LpF3H, which had consistent expression patterns with their upstream and downstream metabolites, and there is a significantly positive correlation between them. Compared to mature leaves, stems and young leaves are higher in the expression levels of key structural genes. We deduced that the MYB and bHLH transcription factors regulated the biosynthesis of different flavonoid metabolites and their regulatory patterns. Among them, LpMYB2, LpMYB20, LpMYB54, LpMYB12, and LpWD40-113 positively regulated the biosynthesis of flavones and flavanones. This discovery preliminarily revealed the pathways and key genes of flavonoid biosynthesis in L. polystachyus, which provided a reference for further study on flavonoid biosynthesis.

Transcriptome and Metabolome Analysis of the Synthesis Pathways of Allelochemicals in Eupatorium adenophorum.

Eupatorium adenophorum (Crofton weed) is an invasive weed in more than 30 countries. It inhibits the growth of surrounding plants by releasing allelochemicals during its invasion. However, the synthetic pathways and molecular mechanisms of its allelochemicals have been rarely reported. In this study, the related genes and pathways of allelochemicals in E. adenophorum were analyzed. Transcriptome analysis showed that differentially expressed genes (DEGs) were mainly enriched in the phenylpropanoid biosynthetic pathway and flavonoid biosynthetic pathway. Thirty-three DEGs involved in the synthesis of allelochemicals were identified, and 30 DEGs showed significant differences in blades and stems. Six allelochemicals were identified from blades and stems by ultraperformance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS). Correlation analysis of genes and metabolites showed a strong correlation between the five genes and allelochemicals. In addition, this study supplemented the biosynthetic pathway of Eupatorium adenophorum B (HHO). It was found that acyclic sesquiterpene synthase (NES), δ-cadinene synthase (TPS), and cytochrome P450 (P450) were involved in the synthesis of HHO. These findings provide a dynamic spectrum consisting of allelochemical metabolism and a coexpression network of allelochemical synthesis genes in E. adenophorum.

A systematic review of newborn and childhood hearing screening around the world: comparison and quality assessment of guidelines.

BACKGROUND: This study aimed to assess the quality of global guidelines or consensus statements for newborn and childhood hearing screening, as well as to compare various guidelines between other countries and China. METHODS: A PROSPERO registered systematic review (number CRD42021242198) was conducted. Multiple electronic databases and government websites including PubMed, EMBASE, Web of Science, CENTRAL, Cochrane Library, and BMJ Best Practice were searched from inception until May 2021. The latest national and international guidelines, consensus statements, technical specifications, and recommendations regarding newborn or childhood hearing screening that were published in Chinese or English medical journals or elsewhere with the full version available online. The following information was extracted independently by two reviewers for comparative analysis: titles, authors, publication year, country, the source organization, and main key recommendations using systems for assigning the level of evidence and strength of recommendations. The quality of the guidelines was assessed by three independent reviewers using the Appraisal of Guidelines for Research and Evaluation, 2nd edition. Intraclass correlation coefficients (ICCs) were calculated to assess among-reviewer agreement. RESULTS: We assessed 15 newborn and 6 childhood hearing screening guidelines, respectively. Most newborn guidelines recommend the 1-3-6 guidelines and pre-discharge screening; however, the specific screening times differ. 93.33% of newborn hearing guidelines recommend "primary screening-re-screening-diagnosis-intervention" for well-babies while 73.33% of the guidelines recommend "initial screening-diagnosis-intervention" for newborns in neonatal intensive care unit (NICU); 33.33% of the newborn hearing guidelines recommended initial screening coverage of > 95% while 46.66% did not mention it. Further, 26.66% of the newborn hearing guidelines recommended a referral rate to diagnosis within 4% while 60% did not mention it. Regarding childhood hearing screening guidelines, the screening populations differed across guidelines (age range: 0-9 years); most guidelines recommend pediatric hearing screening for all preschoolers. Only 50% of the guidelines specify screening and re-screening techniques, including pure-tone hearing screening, OAE, tympanometry, and others. The "Clarity of Presentation" domain achieved the highest mean score, and the lowest was "Editorial Independence" both in newborn and childhood guidelines. Overall score of newborn hearing screening guidelines ranged from 3 (2018 Europe) to 7 (2019 America), with an average score of 5.33. Average score of childhood hearing screening guidelines was 4.78, with the score ranging from 4 (2017 England, 2012 Europe, 2016 WHO) to 6.67 (2011 America). ICC analysis revealed excellent agreement across 21 guidelines (> 0.75). CONCLUSIONS: These findings indicated newborn hearing screening guidelines had superior quality over childhood ones. Comparative analysis suggested that recommendations of the Chinese newborn and pediatric hearing screening protocols are consistent with the mainstream international opinion. Moreover, this analysis demonstrated that "Editorial Independence" and "Stakeholder Involvement" have the greatest opportunities for improvement. These results may help to advance the quality of hearing screening guidelines in clinical practice and guide evidence-based updates.

[Current status of newborn deafness gene screeningin parts of China].

Objective:To analyze the current status of newborn deafness gene screening from 2016 to 2017 in multiple regions of China, and to provide a reference for further promotion and application. Method:The "newborn deafness gene screening questionnaire" was sent to 41 institutions in eastern, central and western China after expert demonstration. The survey content included status of genetic screening, screening methods, the number of screenings, and the status of positive detections from January 1st, 2016 to December 31th, 2017. Each institution returned the questionnaire, the investigator conducted data verification and quality sampling. Finally, we performed analysis of screening methods and the positive detection rate of each gene on questionnaires with complete data. Result:Forty-one questionnaires were sent out and 41 were returned, the questionnaire return rate was 100%, in which 12 questionnaires were complete. Of the 41 institutions, 15 carried newborn deafness gene screening, with a rate of 36.59%（15/41）. The highest rate was in the east（72.22%, 13/18）, and the differences among the regions were statistically significant. As for the screening methods, among 12 questionnaires with complete data, 9 variants in 4 genes and 20 variants in 4 genes accounted for the highest proportion, both with the rate of 33.33%（4/12）, followed by 15 variants in 4 genes（25%, 3/12） and 5 variants in 3 genes（8.34%, 1/12）. A total of 340, 521 neonates were included in the study, and 17, 036 were positive for screening, with a positive rate of 5.00%. Among them, the single heterozygous mutation rate of GJB2 gene was 2.43%（8269/340, 521）, the biallele mutation rate was 0.02%（56/340, 521）,the single heterozygous mutation rate of SLC26A4 gene was 1.99%（6771/340, 521）, the biallele mutation rate was 0.01%（39/340, 521）,the single heterozygous mutation rate of GJB3 gene was 0.33%（1140/340, 521）, the mitochondrial 12SrRNA gene mutation rate was 0.22%（746/340, 521） and the double-gene heterozygous mutation rate was 0.004%（15/340, 521）. Conclusion:From 2016 to 2017, the newborn deafness gene screening is more extensive in the eastern region of China than in the central and western regions. In institutions that have carried out deafness gene screening, 9 variants in 4 genes and 20 variants in 4 genes are widely used; the GJB2 gene and SLC26A4 gene mutations are the most common. The results could provide references for areas where deafness gene screening is about to be performed.

Current status of universal newborn hearing screening program at 26 institutions in China.

OBJECTIVES: The present study aimed to determine the status of a universal newborn hearing screening (UNHS) program being conducted in parts of China, by comparing differences in the program findings between 2016 and 2017, as well as across regions in China. METHODS: This study investigated a nationally representative sample of newborns from 26 provinces, autonomous regions, and municipalities in mainland China. A ''Newborn Hearing Screening Survey'' questionnaire was sent to 43 hearing screening institutions throughout China and the data were analyzed, with appropriate quality control throughout the study process. RESULTS: Twenty-six questionnaires, covering 55.88% (19/34) of the provincial administrative regions in China were appropriately completed. The overall sampling frame comprised 238,795 (year 2016) and 229,185 (year 2017) newborns, respectively. We found differences between two years, the initial screening coverage in 2017 (96.10%) was higher than that in 2016 (94.96%); the referral rate at initial screening in 2017 (9.21%) was lower than that in 2016 (10.26%); and the rescreening rate in 2017 (73.50%) was higher than that in 2016 (68.44%). We found differences across three regions, the rescreening rate were highest in West China, the referral rate at rescreening and the referral rate to diagnostic audiological assessment diagnosis were both highest, while the hearing-loss rate was lowest, in the East China in two years. Overall, 61.54% (n = 16) reported using otoacoustic emissions (OAEs), while 38.46% (n = 10) reported using OAEs in combination with automated auditory brainstem response (AABR) tests, for the initial screening. For rescreening, most sites (n = 19, 73.08%) reported using OAEs in combination with AABR, followed by OAEs only (n = 4, 15.38%) and AABR only (n = 3, 11.54%). Of the twenty-six institutions, 57.69% (n = 15) were equipped with a digital information management system for UNHS program, East China had the highest rate of it (81.82%, 9/11). CONCLUSIONS: This study indicated that implementation of a UNHS program had essentially been achieved in many regions of China under the guidance of technical specifications for newborn hearing screening. Compared with 2016, the overall quality of the UNHS program had improved in 2017 and that in East China was better than in the Midland and West China. However, national quality control of the UNHS program is still required to enhance the quality of the program and public education needs to be emphasized to improve the rescreening and reception rate.

[Analysis of genotypes and hearing phenotypes of mutation infants with deafness].

Objective:The aim of this study is to explore the genotype and hearing phenotype of deaf infants with mutation of GJB2 gene. Method:Subjects were 121 infants with GJB2 gene mutations who were treated in the Children's Hearing Diagnosis Center of Beijing Tongren hospital. All subjects were accepted to undertake the universal newborns hearing screening（UNHS） and series of objective audiometry, including auditory brainstem response, distortion product otoacoustic emission, auditory steady-state response and other audiological tests. All subjects were screened for nine pathogenic variants in four genes or all exons of the GJB2 gene, and then were diagnosed as infants with GJB2 gene mutations. Initially, analyzing their genotypes and hearing phenotypes generally. Then, the subjects were divided into two groups according to the genotypes: T/T group（truncated/truncated mutations, 89 cases） and T/NT group（truncated/non-truncated mutations, 32 cases）. Chi-square test was used to analyze the results of UNHS, hearing degree, audiogram patterns and symmetry/asymmetry of binaural hearing phenotype. Eventually, analyzing the results of UNHS. Result:The most common truncated mutation was c.235delC（64.88%, 157/242） and the most common non-truncated mutation was c.109G>A（11.16%, 27/242）. The homozygous mutation of c.235delC/c.235delC was the dominant in T/T group（38.84%, 47/121）, and the compound heterozygous mutation of c.235delC/c.109G>A was the dominant in T/NT group（18.18%, 22/121）. 81.82%（99/121） of subjects failed in UNHS, including 74.38%（90/121） with bilateral reference, 7.44%（9/121） with a single pass. The refer rate of UNHS of group T/T and T/NT were 86.52%（77/89） and 68.75%, respectively. There was a statistically significant difference between the two groups（P<0.05）. 85.95%（104/121） of subjects were diagnosed as hearing loss and 14.05%（17/121） of subjects were diagnosed as normal hearing. The degree of hearing loss: profound, severe, moderate and mild were 31.40%（38/121）, 19.01%（23/121）, 24.79%（30/121） and 10.74%（13/121）, respectively. There was no subjects with normal hearing in T/T group and individuals with severe and profound hearing loss accounted for the highest proportion（65.17%, 58/89）, while in T/NT group, normal hearing accounted for 53.13%（17/32） and mild and moderate hearing loss accounted for the highest proportion（37.5%, 12/32）. There was statistically significant difference between the two groups（P<0.05）. Of 104 patients（208 ears） with hearing loss, the audiogram patterns: flat, descending, ascending, residual, Valley and other types were 49.03%（102/208）, 12.02%（25/208）, 8.65%（18/208）, 7.69%（16/204）, 3.36%（7/204） and 19.23%（40/204）, respectively. The two most common types in T/T group were flat（47.19%, 84/178） and other types（20.22%, 36/178）, while in T/NT group were flat（60.00%, 18/30） and ascending（20.00%, 6/30）. There was statistically significant difference between the two groups（P<0.05）. There were 50 cases（48.07%） with symmetrical hearing phenotype and 54 cases（51.93%） with asymmetrical hearing phenotype. Asymmetry was predominant in T/T group（53.93%, 48/89）, and symmetry was predominant in T/NT group（60.00%, 9/15）. There was no statistically significant difference between the two groups（P>0.05）. Conclusion:In this study, c.235delC/c.235delC homozygous mutation was dominant in T/T group and c.235delC/c.109G>A heterozygous mutation was dominant in T/NT Group. The hearing phenotypes in T/T group were mostly bilateral asymmetric severe hearing loss, and those in T/NT Group were bilateral symmetric mild to moderate hearing loss, special attention should be paid to the audiological characteristics of different genotypes.

Mutation analysis of the SLC26A4 gene in three Chinese families.

In order to investigate the genetic causes of hearing loss in a Chinese proband (in Family A) with enlarged vestibular aqueduct (EVA) and to investigate the genotype of two Chinese probands with SLC26A4 singe-allelic mutation and normal hearing (in Families B and C, respectively), the three probands and their parents were clinically and genetically evaluated. Twenty exons and flanking splice sites of the SLC26A4 gene were screened for pathogenic mutations via amplification with PCR and bidirectional sequencing. As controls, a group of 400 healthy newborns from the same ethnic background underwent SLC26A4 gene screening using the same method. The three probands all harbored two mutations in the SLC26A4 gene in the form of compound heterozygosity. The genotypes of mutations in Families A, B, and C are c.1211C>A/c.919-2A>G, c.1729G>A/c.919-2A>G, and c.1286C>A/c.919-2A>G, respectively. The missense mutations c.1211C>A (p.T430Q) in exon 10 and c.1729G>A (p.V577I) in exon 16 are both reported for the first time and were absent in 400 healthy newborns. c.1211C>A has Glutamine (Gln) at amino acid 430 instead of Threonine (Thr), and c.1729G>A has Isoleucine (Ile) at amino acid 577 instead of Valine (Val). c.1286C>A, a mutation previously reported in DVD and HGMD, was associated with Mondini deformity, but a proband with the c.1286C>A mutation in this study was normal. This study has demonstrated that the novel missense mutation c.1211C>A in compound heterozygosity with c.919-2A>G in the SLC26A4 gene is likely to be the cause of deafness in Family A. A novel variant, c.1729G>A, was identified and is likely benign. The pathogenicity of the c.1286C>A mutation warrants more in-depth study. These findings will broaden the spectrum of known SLC26A4 mutations in the Chinese population, providing more information for genetic counseling and diagnosis of hearing loss with EVA.

Analysis of mutations in the FOXI1 and KCNJ10 genes in infants with a single-allele SLC26A4 mutation.

The current study investigated how the FOXI1 and KCNJ10 genes were affected in infants with a single-allele mutation in the SLC26A4 gene, and it determined the audiological phenotypes of infants with double heterozygous mutations (DHMs) in the three genes. Subjects were 562 infants with a single-allele SLC26A4 mutation detected during neonatal deafness genetic screening; the infants were seen as outpatients by Otology at Beijing Tongren Hospital. All subjects underwent SLC26A4 sequencing. Twenty infants had a second-allele variant while the remaining 542 had an SLC26A4 single-allele mutation. Infants also underwent FOXI1 and KCNJ10 sequencing. All patients with double heterozygous mutations in the aforementioned genes underwent an audiological evaluation and a limited imaging study; variants and audiological phenotypes were analyzed. Of 562 patients, 20 had SLC26A4 bi-allelic mutations; 8 carried single mutations in both SLC26A4 and KCNJ10. No pathogenic mutations in the FOXI1 gene were found. Four missense mutations in KCNJ10 were detected, including c.812G>A, c.800A>G, c.53G>A, and c.1042C>T. Eight individuals with a DHMs all passed universal newborn hearing screening, and all were found to have normal hearing. These data suggest that individuals with an SLC26A4 single-allele mutation, combined with FOXI1 or KCNJ10 gene mutations, do not suffer hearing loss during infancy, though this finding is worthy of further follow-up and in-depth discussion.

Genotyping and audiological characteristics of infants with a single-allele SLC26A4 mutation.

OBJECTIVES: To identify second-allele variant in infants with a known single-allele mutation of the SLC26A4 gene and to determine the frequency of their occurrence; and to investigate the clinical audiological characteristics of infants with bi-allelic mutations in SLC26A4. METHODS: The study subjects were 371 patients with a single-allele SLC26A4 mutation detected by neonatal deafness gene screening (4 genes and 9 pathogenic variants) who were treated at the otology outpatient department of Beijing Tongren Hospital. The exonic and flanking splice site regions of the SLC26A4 gene were sequenced for all patients. All patients with bi-allelic SLC26A4 mutations underwent audiological evaluation, and some also underwent temporal bone computed tomography and/or inner ear magnetic resonance imaging. RESULTS: Of the 371 patients, 314 (84.64%) had an c.919-2A > G heterozygous mutation and 57 (15.36%) had a c.2168A > G (p.H723R) heterozygous mutation. 13 patients (3.50%) had a second-allele variant, including 11 (2.96%) with pathogenic mutations and 1 (0.27%) with a likely benign variant. Of the 13 patients with bi-allelic mutations, 11 had hearing loss and 2 had normal hearing, the latter of whom had c.919-2A > G/c.1766A > G and c.919-2A > G/c.757A > G compound heterozygous mutations, respectively. Four of the 13 patients with bi-allelic mutations had passed the universal newborn hearing screening, including 2 cases (15.38%) with hearing loss. The most prevalent degree of hearing loss was profound (40.91%), followed by severe (36.36%). The most prevalent audiometric configuration was sloping hearing loss (50.00%), followed by flat-type hearing loss (40.91%). CONCLUSIONS: This is the first report in China of the frequency of occurrence of second-allele variant in infants with a known single-allele mutation of the SLC26A4 gene; the frequency was 3.50% for any type of variant and 2.96% for pathogenic mutations. A novel variant, c.1766A > G (p.Q589R), which is likely benign, was identified. The pathogenicity of c.757A > G (p.I253V) mutation deserves more in-depth research. For infants with bi-allelic SLC26A4 mutations, the degree of hearing loss was mainly severe-to-profound and the audiometric configuration was mainly sloping.

The higher serum endocan levels may be a risk factor for the onset of cardiovascular disease: A meta-analysis.

OBJECTIVE: Endothelial dysfunction was widely regarded as the initial lesion in the multifactorial pathogenesis of cardiovascular disease (CVD). Serum endocan, a novel endothelial dysfunction biochemical marker, is involved in the development of CVD. Here, we fulfilled a meta-analysis to evaluate the association between CVD and serum endocan levels. METHOD: The relevant published literature was searched through large literature databases, including PubMed, Embase, Cochrane Library, SinoMed, and Web of Science, up to June 1, 2018. The data were extracted from the studies. Stata software was used to perform a meta-analysis. RESULT: Fifteen original studies with a total of 1839 patients and 1258 controls fulfilled the inclusion criteria and were included in the study dataset. Meta-analysis showed that the levels of serum endocan in patients with hypertension, coronary artery disease, and coronary slow flow were higher than those in the control group. The pooled standardized mean differences and 95% confidence intervals of endocan concentrations in those 3 groups were 0.53 [0.19-0.86], P < .01; 0.99 [0.51-1.39], P < .01; and 0.62 [0.45-0.78], P < .01, respectively. Further analysis showed that the level of serum endocan in hypertension patients with coronary artery disease was higher than that in patients with hypertension (0.61 [0.30-0.92], P < .01). Sensitivity analysis and subgroup analysis were use to confirm the above results. CONCLUSIONS: In this meta-analysis, we further confirmed that serum endocan level was significantly increased in the CVD population. The high serum endocan level may be one of the risk factors for CVD.

Novel compound heterozygous mutations in SLC26A4 gene in a Chinese family with enlarged vestibular aqueduct.

In order to investigate the genetic causes of hearing loss in a Chinese proband with nonsyndromic hearing loss and enlarged vestibular aqueduct (EVA), we conducted clinical and genetic evaluations in a deaf proband and her parents with normal hearing. 20 exons and flanking splice sites of the SLC26A4 gene were screened for pathogenic mutations by PCR amplification and bidirectional sequencing. As a control, a group of 400 healthy newborns from the same ethnic background were subjected to SLC26A4 gene screening using the same method. The proband harbored two mutations in the SLC26A4 gene in the form of compound heterozygosity. She was found to be heterozygous for a novel mutation c.574delC (p.Leu192Ter) in exon 5 and for the known mutation c.919-2A>G(c.IVS7-2A>G). Her mother was a heterozygous carrier of the c.919-2A>G mutation, and her father was a heterozygous carrier of the c.574delC and therefore co-segregated with the genetic disease. The c.574delC mutation was absent in 400 healthy newborns. The frameshift mutation causes the leucine (Leu) at amino acid position 192 to become a termination codon, leading to termination of protein sequence coding. This study demonstrates that the novel frameshift mutation c.574delC (p.Leu192Ter) in compound heterozygosity with c.919-2A>G in the SLC26A4 gene is the main cause of deafness in a family. Our study will expand the spectrum of known SLC26A4 mutations in the Chinese population, providing more information on genetic counseling, and diagnosis in hearing loss with EVA.

Children with GJB2 gene mutations have various audiological phenotypes.

The current study retrospectively investigated variations in audiological phenotypes in children with GJB2 gene mutations. Subjects were 128 infants and young children who were seen as outpatients by Otology at Beijing Tongren Hospital from 2012 to 2018. Of the 128 subjects, 99 had biallelic truncating (T/T) mutations and 29 had truncating/nontruncating (T/NT) mutations. Genotypes, results of universal newborn hearing screening (UNHS), and the degree and symmetry of hearing loss were examined in the two groups. Twenty-two subjects (20.37%, 22/128) passed UNHS, including 13 children with T/T mutations and 9 with T/NT mutations. Of the 128 subjects, 22 had normal hearing, 2 had unilateral hearing loss, and 115 had bilateral hearing loss. Severe-to-profound hearing loss was the most prevalent phenotype in children with T/T mutations (73.23%), while normal hearing was prevalent in children with T/NT mutations (41.38%). Symmetrical hearing loss was the main phenotype in both groups, and the number of subjects with symmetrical hearing loss did not differ significantly between the two groups. Therefore, children with GJB2 gene mutations have phenotypic variability in terms of their results of UNHS and their degree and symmetry of hearing loss. Subjects with T/NT mutations of the GJB2 gene were more likely to pass UNHS and had milder hearing loss compared to those with T/T mutations. Symmetrical hearing loss was the main phenotype in the two groups, but 36.53% of children had bilateral asymmetric hearing loss. Parents of all subjects with sensorineural hearing loss were informed that their children may have a GJB2 mutation.