Novel IARS1 variants cause syndromic developmental disorder with epilepsy in a Chinese patient and the literature review.

BACKGROUND: Isoleucinyl-tRNA synthetase (IARS) is encoded by the IARS1 gene and catalyzes the binding of isoleucine to specific tRNA. OBJECTIVE: This study aims to investigate the pathogenicity of novel IARS1 variants and the genotype-phenotype association, in order to expand the spectrum of pathogenic variants and phenotypes of IARS1-related disease and provide new evidence for the phenotypic spectrum of IARS1 variants. METHODS: Clinical data of the proband were collected, and trio whole-exome sequencing (WES) was performed on the proband and the parents. Candidate variants were validated using Sanger sequencing. Bioinformatics software was utilized to analyze the functional consequences of identified variants and predict their potential deleteriousness. RESULTS: A 17-month-old female patient presented with microcephaly, left external ear malformation, decreased muscle strength and tone in all limbs, epileptic seizures, global developmental delay, and developmental regression. Trio WES identified compound heterozygous variants in the IARS1 gene, c.120-1G>A and c.2164C>A, which were novel pathogenic and likely pathogenic variants, respectively. The phenotype of developmental regression has not been reported before. Only one patient with IARS1 compound heterozygous variants has been reported in the world to have an epileptic phenotype, and this is the second patient with an epileptic phenotype. Bioinformatics analysis revealed that the splicing variant disrupted the canonical splice donor site, while the missense variant altered the local electrostatics of the IARS1 protein surface, potentially leading to functional abnormalities. CONCLUSION: This study identified novel IARS1 variants and the phenotype of developmental regression, expanding the spectrum of pathogenic variants and phenotypes of IARS1-related diseases and providing new evidence for the rare phenotype of epileptic seizures caused by IARS1 variants.

[Genetic analysis of a child with Complex cortical dysplasia with other brain malformations type 6 due to a p.M73V variant of TUBB gene].

OBJECTIVE: To explore the genetic basis for a child with multiple malformations. METHODS: A child who had presented at Shanxi Provincial Children's Hospital in February 2021 was selected as the study subject. Clinical data of the patient was collected, and whole exome sequencing (WES) was carried out to screen pathogenic variants associated with the phenotype. Candidate variant was validated by Sanger sequencing of her family members. RESULTS: The child had normal skin, but right ear defect, hemivertebral deformity, ventricular septal defect, arterial duct and patent foramen ovale, and separation of collecting system of the left kidney. Cranial MRI showed irregular enlargement of bilateral ventricles and widening of the distance between the cerebral cortex and temporal meninges. Genetic testing revealed that she has harbored a heterozygous variant of NM_178014.4: c.217A>G (p.Met73Val) in the TUBB gene, which was unreported previously and predicted to be likely pathogenic based on the guidelines from the American College of Medical Genetics and Genomics (ACMG). The child was diagnosed with Complex cortical dysplasia with other brain malformations 6 (CDCBM6). CONCLUSION: CDCBM is a rare and serious disease with great genetic heterogeneity, and CDCBM6 caused by mutations of the TUBB gene is even rarer. Above finding has enriched the variant and phenotypic spectrum of the TUBB gene, and provided important reference for summarizing the genotype-phenotype correlation of the CDCBM6.

Correlation of TGF-ß signaling pathway gene polymorphisms with unexplained recurrent spontaneous abortion.

BACKGROUND: The association of key genes in the transforming growth factor-ß (TGF-ß) signaling pathway and their gene polymorphisms with unexplained recurrent spontaneous abortion (URSA) is unclear. OBJECTIVE: To investigate the association of gene polymorphisms related to the TGF-ß signaling pathway in URSA women. METHODS: The study population consisted of 80 women with URSA and 90 normal control women, of which 10 women with URSA and 10 normal control women underwent high-throughput sequencing to select loci, and the remaining 70 women with URSA and 80 normal control women underwent flight mass spectrometry experiments to verify gene loci polymorphism. A total of 7 polymorphic loci in interleukin-6 (IL-6), TGF-ß1, TNF-α, SMAD1, and TNFRSF4 genes were screened by high-throughput sequencing combined with a review of databases. An SNP flight mass spectrometer (Mass ARRAY detection system) was applied to detect the polymorphisms and their frequencies in 70 women with URSA and 80 normal control women at the 7 gene loci. RESULTS: Among the 7 loci of IL-6, TGF-ß1, TNF-α, SMAD1, and TNFRSF4 genes, 2 loci were found to have significantly different allele and genotype frequency distributions between the 70 URSA and 80 normal controls, one was the IL-6 gene -174G/C locus (rs1800795), the risk of disease was 2.636 and 3.231 times higher in individuals carrying the C allele and CC genotype than in those carrying the G allele and GG genotype, respectively; the other was the TGF-ß1 gene -509T/C locus (rs1800469), and the risk of disease was 1.959 and 3.609 times higher in individuals carrying the T allele and TT genotype than in those carrying the C allele and CC genotype, respectively. The remaining 5 genetic loci have no statistically significant. CONCLUSION: IL-6 gene -174G/C locus (rs1800795) genotype CC and allele C may be the causative factor of URSA, TGF-ß1 gene -509T/C locus (rs1800469) genotype TT and allele T may be the causative factor of URSA, and polymorphisms of the 2 loci may be associated with URSA.

Prenatal diagnosis in a fetuses with a clenched hands, overlapping fingers, and clubfoot due to MED12 deficiency in three affected siblings: A case report.

A fetal clenched hand with overlapping fingers is more common in aneuploidy syndrome and was not well-documented in MED12 deficiency. This study reports the clinical and genetic findings of three affected siblings from a Chinese family. The chromosome karyotype analysis diagram shows that karyotypes of the three children were normal. Trio whole-exome sequencing and Sanger sequencing verification found that there was a MED12 R296Q variant in normal mothers and their two offspring. A pattern of clenched hand with overlapping fingers (clinodactyly) and clubfoot was found in all the three affected siblings by three-dimensional ultrasound. The discovery of this case shows that even if the chromosome karyotype is normal, comprehensive prenatal genetic diagnosis is required when the ultrasound results show a clenched hand with clinodactyly and clubfoot symptoms.

A first-order kinetic model for simulating the aerobic degradation of municipal solid waste.

Aerobic degradation models are important tools for investigating the aerobic degradation behavior of municipal solid waste (MSW). In this paper, a first-order kinetic model for aerobic degradation of MSW was developed. The model comprehensively considers the aerobic degradation of five substrates, i.e., holocellulose, non-cellulosic sugars, proteins, lipids and lignin. The proportion ranges of the five substrates are summarized with the recommended values given. The effects of temperature, moisture content, oxygen concentration and free air space (FAS) on the reaction rates are considered, and the effect of settlement is accounted for in the FAS correction function. The reliability of the model was verified by comparing simulations of the aerobic degradation of low food waste content (LFWC-) and high food waste content (HFWC-) MSWs to the literature. Afterwards, a sensitivity analysis was carried out to establish the relative importance of aeration rate (AR), volumetric moisture content (VMC), and temperature. VMC had the greatest influence on the aerobic degradation of LFWC-MSW, followed by temperature and then AR; for HFWC-MSW, temperature was the most important factor, then VMC and last was AR. The degradation ratio of LFWC-MSW can reach 98.0% after 100 days degradation under its optimal conditions (i.e., temperature: 55 °C, VMC: 40%, AR: 0.16 L min-1 kg-1 DM), while it is slightly higher as 99.5% for HFWC-MSW under its optimal conditions (i.e., temperature: 55 °C, VMC: 40%, AR: 0.20 L min-1 kg-1 DM).

Diagnostic value of CBCT in Chinese children with adenoid hypertrophy.

Objective: The main objectives of the study were to investigate the reliability and accuracy of cone-beam computed tomography (CBCT) in the diagnosis of adenoid hypertrophy in Chinese children and to evaluate its value in clinical diagnosis. Methods: From January 2019 to January 2020, 300 children with sleep snoring in Xinhua Hospital Affiliated to Medical College of Shanghai Jiaotong University were retrospectively studied. All patients underwent nasopharyngoscopy (NE) and CBCT scanning. The sensitivity, specificity, positive predictive value, negative predictive value, positive likelihood ratio, and negative likelihood ratio of CBCT were determined according to the diagnostic criteria of NE, and the consistency between CBCT and NE was evaluated. Results: The clinical study of 300 children patients found that compared with NE, CBCT had a sensitivity of 87.3%, specificity of 89.2%, the positive predictive value of 93.20%, the negative predictive value of 80.5%, the positive likelihood ratio of 8.08, the negative likelihood ratio of 0.14, and Kappa value of .748. Conclusion: CBCT is a reliable and accurate tool for the diagnosis of adenoid hypertrophy and can be used as an alternative examination method for children with contraindications or intolerance during NE. Level of Evidence: 4.

Potential vestibular pathway impairment in children with recurrent vertigo: An investigation through air-conducted sound and galvanic vestibular stimulation-triggered vestibular evoked myogenic potentials.

Objective: This study aims to investigate the potential vestibular pathway impairment through vestibular evoked myogenic potentials (VEMPs) and to explore the pathophysiological significance of these instrument-based findings in children with recurrent vertigo. Materials and methods: The clinical data of 21 children (mean age 4.67 ± 1.39 years) diagnosed as RVC who met the inclusion criteria of the Bárány Society and 29 healthy children (mean age 4.83 ± 1.34 years) enrolled as the control group from February 2021 to December 2021 were collected and analyzed retrospectively. All the subjects underwent both cervical VEMP (cVEMP) and ocular VEMP (oVEMP) triggered by air-conducted sound (ACS) and galvanic vestibular stimulation (GVS), respectively. The elicit rate, latency, and amplitude asymmetry ratio (AAR) of ACS-cVEMP, ACS-oVEMP, GVS-cVEMP, and GVS-oVEMP were analyzed. Results: (1) The elicit rates of ACS-cVEMP and ACS-oVEMP were similar in the two groups (P > 0.05), as well as GVS-cVEMP and GVS-oVEMP (P > 0.05). (2) P1 and N1 latencies of ACS-cVEMP and GVS-cVEMP in the RVC group were longer than those in the control group (P < 0.05). (3) The N1 latency of ACS-oVEMP in the RVC group was shorter than that in the control group (P < 0.05), while there was no significant difference in the P1 latency of ACS-oVEMP (P > 0.05). The N1 and P1 latencies of GVS-oVEMP were not significantly different (P > 0.05). (4) There was no statistical difference in the AAR of ACS-cVEMP and GVS-cVEMP. Although there was an increased AAR of ACS-oVEMP in the RVC group (P < 0.05), the AAR was within the normal range. However, no statistical difference was found in the AAR of GVS-oVEMP in the two groups (P > 0.05). Conclusion: The latencies of ACS-cVEMP and GVS-cVEMP in children with recurrent vertigo were significantly prolonged compared with those in healthy children, and there was no difference in elicit rates of ACS-cVEMP and GVS-cVEMP, suggesting that there might be potential impairment in the inferior vestibular nerve and the subsequent nerve conduction pathway in RVC.

Clinical characteristics of vestibular evoked myogenic potentials in children with recurrent vertigo of childhood.

OBJECTIVE: To explore the possible pathogenesis of recurrent vertigo of childhood (RVC) and the clinical diagnosis value of vestibular-evoked myogenic potentials (VEMPs). METHODS: The clinical data of 19 children (5.95 ± 0.38 years) diagnosed with RVC and 17 normal children (5.35 ± 0.31 years) enrolled in the control (NC) group from April 2017 to February 2021 was collected and analyzed. All subjects were tested for both cervical VEMP (cVEMP) and ocular VEMP (oVEMP). The elicit rate, thresholds, N1 latency, P1 latency, interval, amplitude, and amplitude asymmetry ratio (AAR) of VEMPs were compared and analyzed between the two groups. RESULTS: (1) The elicit rates of cVEMP and oVEMP have no significant difference between the two groups (P > 0.05). (2) The thresholds of cVEMP and oVEMP in the RVC group were higher than that in the NC group (P < 0.05). (3) The N1 latency of cVEMP in the RVC group was longer than that in the NC group (P < 0.05). The P1 latency of cVEMP and latencies of oVEMP have no significant difference between the two groups (P > 0.05). (4) The interval of cVEMP in the RVC group was longer than that in the NC group (P < 0.05), while the interval of oVEMP has no significant difference between the two groups (P > 0.05). (5) The amplitude of cVEMP in the RVC group was higher than that in the NC group (P < 0.05), while the amplitude of oVEMP was similar in the two groups (P > 0.05). (6) The AAR values of oVEMP and cVEMP were similar in the two groups (P > 0.05). CONCLUSIONS: The elicit rates of VEMPs in children with RVC did not differ from that of normal children, but the thresholds were all increased, suggesting reduced sensitivity of the otolith organ and vestibular nerve conduction pathways. The P1 latency of cVEMP was normal in children with RVC, but N1 latency and interval of cVEMP were increased, we finally reached a conclusion that there might be potential impairment in the inferior vestibular nerve and the subsequent nerve conduction pathway in RVC.

Pseudo-Temporal Analysis of Single-Cell RNA Sequencing Reveals Trans-Differentiation Potential of Greater Epithelial Ridge Cells Into Hair Cells During Postnatal Development of Cochlea in Rats.

The hair cells of the cochlea play a decisive role in the process of hearing damage and recovery, yet knowledge of their regeneration process is still limited. Greater epithelial ridge (GER) cells, a type of cell present during cochlear development that has the characteristics of a precursor sensory cell, disappear at the time of maturation of hearing development. Its development and evolution remain mysterious for many years. Here, we performed single-cell RNA sequencing to profile the gene expression landscapes of rats' cochlear basal membrane from P1, P7, and P14 and identified eight major subtypes of GER cells. Furthermore, single-cell trajectory analysis for GER cells and hair cells indicated that among the different subtypes of GER, four subtypes had transient cell proliferation after birth and could transdifferentiate into inner and outer hair cells, and two of them mainly transdifferentiated into inner hair cells. The other two subtypes eventually transdifferentiate into outer hair cells. Our study lays the groundwork for elucidating the mechanisms of the key regulatory genes and signaling pathways in the trans-differentiation of GER cell subtypes into hair cells and provides potential clues to understand hair cell regeneration.

Overexpression of c-Jun inhibits erastin-induced ferroptosis in Schwann cells and promotes repair of facial nerve function.

Myelin undergoes various changes after nerve injury, and c-Jun has a close relationship with Schwann cells (SCs). However, it remains unclear whether c-Jun can be involved in nerve repair by regulating ferroptosis. To explore this, we first set up a facial nerve injury model and detected the changes of ferroptosis-related proteins and c-Jun by immunofluorescence and Western blot. Then, we cultured RSC 96 and pSCs, and studied the potential regulatory relationships by a combination of experimental methods such as CCK-8, ELISA, immunofluorescence, qRT-PCR, Western blot and viral transfection. Finally, we corroborated the role of c-Jun through animal experiments. Our experiments revealed that ferroptosis occurs after facial nerve injury. Erastin decreased GPX4, c-Jun proteins and GSH content, while PTGS2, NRF2, HO-1 proteins, MDA, Fe2+ and ROS contents increased. This effect was inhibited after c-Jun overexpression but was reversed after the addition of c-Jun siRNA. Besides, we proved in vivo that c-Jun could inhibit ferroptosis and promote the recovery of facial nerve function. In conclusion, our study identified the relationship between c-Jun and ferroptosis during peripheral nerve injury repair, which provides new ideas for studying peripheral nerve injury and repair.

DHI Increases the Proliferation and Migration of Schwann Cells Through the PI3K/AKT Pathway and the Expression of CXCL12 and GDNF to Promote Facial Nerve Function Repair.

The facial nerve is one of the vulnerable nerves in otolaryngology. Repair and recovery of facial nerve injury have a high priority in clinical practice. The proliferation and migration of Schwann cells are considered of great significance in the process of nerve injury repair. Danhong injection (DHI), as a common drug for cardiovascular and cerebrovascular diseases, has been fully certified in neuroprotection research, but its role in facial nerve injury is still not clear. Our study found that DHI can promote the proliferation and migration of RSC96 cells, a Schwann cell line, and this effect is related to the activation of the PI3K/AKT pathway. LY294002, an inhibitor of PI3K, inhibits the proliferation and migration of RSC96 cells. Further studies have found that DHI can also promote the expression of CXCL12 and GDNF at gene and protein levels, and CXCL12 is, while GDNF is not, PI3K/AKT pathway-dependent. Animal experiments also confirmed that DHI could promote CXCL12 and GDNF expression and promote facial nerve function recovery and myelin regeneration. In conclusion, our in vitro and in vivo experiments demonstrated that DHI could promote the proliferation and migration of Schwann cells through the PI3K/AKT pathway and increase the expression of CXCL12 and GDNF to promote facial nerve function repair.

Single-Cell RNA Sequencing Analysis Reveals Greater Epithelial Ridge Cells Degeneration During Postnatal Development of Cochlea in Rats.

Greater epithelial ridge cells, a transient neonatal cell group in the cochlear duct, which plays a crucial role in the functional maturation of hair cell, structural development of tectorial membrane, and refinement of audio localization before hearing. Greater epithelial ridge cells are methodologically homogeneous, while whether different cell subtypes are existence in this intriguing region and the degeneration mechanism during postnatal cochlear development are poorly understood. In the present study, single-cell RNA sequencing was performed on the cochlear duct of postnatal rats at day 1 (P1) and day 7 (P7) to identify subsets of greater epithelial ridge cell and progression. Gene ontology and Kyoto Encyclopedia of Genes and Genomes pathway enrichment analysis were used to examine genes enriched biological processes in these clusters. We identified a total of 26 clusters at P1 and P7 rats and found that the cell number of five cell clusters decreased significantly, while four clusters had similar gene expression patterns and biological properties. The genes of these four cell populations were mainly enriched in Ribosome and P13K-Akt signal pathway. Among them, Rps16, Rpsa, Col4a2, Col6a2, Ctsk, and Jun are particularly interesting as their expression might contribute to the greater epithelial ridge cells degeneration. In conclusion, our study provides an important reference resource of greater epithelial ridge cells landscape and mechanism insights for further understanding greater epithelial ridge cells degeneration during postnatal rat cochlear development.

Hearing Protection Outcomes of Analog Electrode Arrays Coated with Different Drug-Eluting Polymer Films Implanted into Guinea Pig Cochleae.

OBJECTIVE: To investigate the hearing protection outcomes of different drug-eluting analog electrode arrays implanted into guinea pig cochleae. METHODS: Sixty guinea pigs were randomly divided into a negative control group and five experimental groups implanted separately with blank (drug carrier), dexamethasone (DXM), aracytine (Ara-C), Ara-C+DXM, and nicotinamide adenine dinucleotide (NAD+) eluting analog electrode arrays. Micro CT was used to supervise the surgical procedure. Auditory brainstem response (ABR) thresholds of the guinea pigs were measured and analyzed. RESULTS AND CONCLUSIONS: Compared with the negative control, all other groups showed a significant increase in ABR threshold (p<0.001) after surgery. Among them, there was no obvious difference between the blank (0 vs 90 days: 59.70±10.57 vs 64.60±9.47 dB SPL) and the NAD+ group (0 vs 90 days: 59.90±9.87 vs 64.70±8.65 dB SPL). On the other hand, the ABR thresholds in the DXM (0 days: 58.10±10.73 dB SPL; 90 days: 51.70±9.07 dB SPL) and the Ara-C group (0 days: 59.00±10.05 dB SPL; 90 days: 51.60±8.48 dB SPL) decreased significantly compared with the former two groups (p<0.001). However, the Ara-C+DXM group showed no further benefit (p>0.05). In addition, a significantly higher survival rate of spiral ganglion neurons in cochleae was observed in the Ara-C and/or DXM groups.

[Research advances on umbilical cord blood derived mesenchymal stem cell in therapy of sensorineural hearing loss].

Sensorineural hearing loss（SNHL） is a difficult and hot issue in the field of otology. The effect of medications targeting the mechanism of SNHL are often unsatisfactory for hearing improvement. Hearing aids and cochlear implants are the mainstream treatment methods at present, but neither of them can reverse the pathological changes of affected inner ear. In recent years, mesenchymal stem cells transplantation has been confirmed by several animal studies and clinical trials to have great potential for clinical applications in restoring the structure of the inner ear and hearing improvement to some extent. In this review, we review the characteristics of umbilical cord blood derived mesenchymal stem cells（UCBMSCs）, the approaches of transplantation, the efficacy and mechanism of UCBMSCs in the treatment of SNHL, and the safety of clinical application, covering the existing problems and future prospects of this rising treatment.

The Correlation of a 2D Volume-Referencing Endolymphatic-Hydrops Grading System With Extra-Tympanic Electrocochleography in Patients With Definite Ménière's Disease.

Background: Although magnetic resonance imaging (MRI) of the membranous labyrinth and electrocochleography (ECochG) have been used to diagnose endolymphatic hydrops (ELH) in patients with Ménière's disease (MD), the relationship between imaging and ECochG is not well-documented. Objectives: This study evaluates the ELH using 3D-FLAIR MRI and extra-tympanic ECochG (ET-ECochG) and correlates the results from 3D-FLAIR MRI to those from ET-ECochG. Materials and Methods: 3D-FLAIR MRI images of 50 patients were assessed using a 2D volume-referencing grading system (VR scores, relative scores according to the known volumes of the cochlea, vestibule, and semicircular canals). Forty healthy subjects were included and compared to 51 definite MD ears of 50 patients while analyzing the ET-ECochG, which used a self-made bronze foil electrode. The amplitude ratio of the summating potential (SP) to the action potential (AP) (SP/AP) and the area ratio of SP to AP (Asp/Aap) were collected. Relative ELH grade scores were then correlated to ET-ECochG (SP/AP, Asp/Aap). Results: The VR scores showed a better correlation (r = 0.88) with the pure tone average (PTA), disease duration, and vertigo frequency of MD than the Bernaerts scores (grading the cochlea and vestibule separately) (r = 0.22). The SP/AP and Asp/Aap of the unilateral MD patients were statistically comparable to those measured in contralateral ears and the results between the definite MD ears with healthy ears were statistically comparable (p < 0.05). In a ROC analysis Asp/Aap (area under curve, AUC 0.98) significantly (p = 0.01) outperformed SP/AP (AUC 0.91). The total score of ELH, vestibular ELH, and cochlear ELH were also correlated with SP/AP and Asp/Aap. The strongest correlation was found between the Asp/Aap and cochlear ELH (r = 0.60). Conclusion: The 2D volume-referencing grading system was more meaningful than the Bernaerts scores. A correlation was found between ELH revealed by 3D-FLAIR MRI and the SP/AP of ET-ECochG in evaluating definite MD patients. The Asp/Aap appeared a more sensitive and reliable parameter than SP/AP for diagnosing the ELH of the membranous labyrinth.

Low Expression of LEFTY1 in Placental Villi Is Associated with Early Unexplained Miscarriage.

OBJECTIVE: To investigate the function and underlying mechanism of transforming growth factor­beta (TGF-ß)/bone morphogenetic protein (BMP) signaling pathway in early unexplained miscarriage. STUDY DESIGN: Expression profiles of genes involved in TGF-ß/BMP signaling pathway were compared between placental villous tissue samples from 2 women with missed abortion and those from 2 women with induced abortion by microarray assay. The protein expression level of the most downregulated gene­LEFTY1­was further measured using western blotting in another 8 women with missed abortion and 7 women with induced abortion. RESULTS: A total of 24 genes showed differential expression level between the 2 groups. Their functions were further investigated, of which 6 of 13 upregulated genes were TGF-ß responsive genes. The most reduced gene is LEFTY1, an antagonist of TGF-ß ligand. The protein expression level of LEFTY1 was confirmed to show the same trend as microarray using western blotting. CONCLUSION: A reduced expression of LEFTY1 in women with missed abortion was identified as com-pared with women with induced abortion, which may result in a dysregulation of TGF-ß signaling and may be the underlying mechanism of missed abortion.

[Phenotypic and genetic analysis of a child featuring multiple malformations due to copy number variation on chromosome 5].

OBJECTIVE: To determine the origin of chromosomal aberration for a child featuring multiple malformation, and to correlate the genotype with phenotype. METHODS: Routine G-banding was performed to analyze the karyotype of the patient and her parents, and array comparative genomic hybridization (array CGH) was used for fine mapping of the aberrant region. RESULTS: The karyotype of the child was ascertained as 46,XY. Array CGH has mapped a 14.21 Mb deletion to 5p15.2p15.33, and a very small 3.67 Mb duplication to 5q35.3. The patient has presented features such as mental retardation, heart defect, low-set ears, hypertelorism and down-slanting palpebral fissures. CONCLUSION: Chromosome 5 copy number variation can cause multiple malformation. In contrast to routine karyotype analysis, array CGH can map aberrant region with much higher resolution and accuracy.