OBJECTIVE: To analyze the carrier rates and profiles of pathogenic and likely pathogenic variants for hearing loss-related genes MYO7A, PCDH15, and CDH23 among neonates in Nanjing city through targeted next-generation sequencing (NGS). METHODS: Heel-prick blood samples were collected from 30 043 newborns delivered at Nanjing Women and Children's Health Care Hospital between March 2022 and April 2024. Dried blood spots were prepared, and genomic DNA was extracted. Targeted NGS was applied to detect variants across the full coding regions of the MYO7A, PCDH15, and CDH23 genes. The carrier rates and profiles of pathogenic and likely pathogenic variants of the three genes were analyzed. This study was approved by the Medical Ethics Committee of Nanjing Maternal and Child Health Care Hospital (Ethics No.: 2021KY-071). RESULTS: The carrier rates of pathogenic and likely pathogenic variants (with ≥ 1 variant site) for the MYO7A, PCDH15, and CDH23 genes were 0.340%, 0.226%, and 0.156%, respectively. A total of 65, 49, and 30 variant types were detected in the MYO7A, PCDH15, and CDH23 genes, respectively. For MYO7A, single base variants were predominant, with the most common variant being c.5581C>T, followed by c.1343+1G>A, c.2837T>G, and c.5660C>T, with allelic frequencies of 0.013% (8/60 086), 0.007% (4/60 086), 0.007% (4/60 086), and 0.007% (4/60 086), respectively. PCDH15 variants were mainly deletions, with the most common variant site being c.4699_4715dupAGAGAAAAGATTCAGAG, followed by c.3441delA, c.440T>G, and c.4733_4736delTCAG, with allelic frequencies of 0.015% (9/60 086), 0.005% (3/60 086), 0.005% (3/60 086), and 0.005% (3/60 086), respectively. For CDH23, single base variants were predominant, with c.6604G>A being the most common, followed by c.6085C>T, c.6050+9G>A, and c.6253+1G>A, with allelic frequencies of 0.013% (8/60 086), 0.012% (7/60 086), 0.005% (3/60 086), and 0.005% (3/60 086). CONCLUSION This study analyzed the carrier rates and profiles of pathogenic and likely pathogenic variants of the MYO7A, PCDH15, and CDH23 genes, which can provide more evidence for the prevention and management of deafness in the region.
Humans ; Cadherins/genetics* ; High-Throughput Nucleotide Sequencing/methods* ; Infant, Newborn ; Female ; Myosin VIIa/genetics* ; Cadherin Related Proteins ; Male ; Hearing Loss/genetics* ; Myosins/genetics* ; Heterozygote

Objective:To understand the regional prevalence and hotspot mutations through analysis of genetic screening results for newborns with pseudohypertrophic muscular dystrophy.Methods:A total of 22 813 newborns (12 065 males and 10 748 females) born at the Women's Hospital of Nanjing Medical University from March 18, 2022, to October 31, 2023, were selected. The Dystrophin gene ( DMD) was detected using chip capture next-generation sequencing technology, followed by bioinformatics analysis. Pathogenic mutations identified were validated using multiplex ligation-dependent probe amplification (MLPA) and Sanger sequencing. Serum creatine kinase levels were also tested in suspected male patients. Descriptive analysis was applied for this study. Results:Among the 10 748 girls, 14 carriers of DMD gene were detected (0.013%), of which, nine cases were validated in the family; one case was a de novo mutation, five were inherited from the mother, and three were inherited from the father. The screening identified nine suspected patients among the boys (0.075%), and eight of them were confirmed by family validation, in which three were de novo mutations, and five were inherited from the mother. Among all identified DMD mutations, deletions were the most common one, accounting for 52.2% (12/23), incluling four cases of deletion at 49-51 exon. Conclusions:Newborn genetic screening based on chip capture next-generation sequencing technology combined with bioinformatics analysis is helpful in early detection of patients and carriers of pseudohypertrophy muscular dystrophy. According to the preliminary statistics, the incidence rate of DMD/BMD in this area is 1/1 341 male infants and the hotspot mutation is exon 49 to 51 deletion.

Objective:To establish the cut-off value of tetradecenoyl carnitine (C14∶1)/dodecenoyl carnitine(C12∶1) based on non-derivatized tandem mass spectrometry (MS/MS), and to explore the application value of C14∶1/C12∶1 to screen newborns for very long chain acyl-CoA dehydrogenase deficiency (VLCADD), determining the best combination of indicators for screening VLCADD.Methods:This retrospective study included data from 17 newborns with VLCADD detected by MS/MS and confirmed by acyl-CoA dehydrogenase very long chain ( ACADVL) gene detection, and 423 507 newborns with normal MS/MS results. The data from these newborns were collected from January 2014 to December 2021 as the newborns received neonatal screening in Nanjing Neonatal Disease Screening Center and Suzhou Neonatal Disease Screening Center. All newborns were divided into 3 groups: all newborns group, full-term newborns group and normal-birth-weight newborns group, and the cut-off values of C14∶1/C12∶1 for VLCADD in these 3 groups were determined by their receiver operating characteristic (ROC) curves individually. With these results, a total of 5 interpretation schemes were composed using different indicators alone or jointly: scheme 1 being C14∶1/C12∶1, scheme 2 being C14∶1, scheme 3 being C14∶1+C14∶1/C2+C14∶1/C16, scheme 4 being C14∶1/C12∶1+C14∶1, and scheme 5 being C14∶1/C12∶1+C14∶1+C14∶1/C2+C14∶1/C16. The detection rate, false-positive rate and positive predictive value of each scheme were calculated, and their screening efficiencies were statistically compared by Chi-square tests. Results:The cut-off values of C14∶1/C12∶1 for VLCADD in the 3 newborn groups were all 2.80. The detection rates of VLCADD with all 5 interpretation schemes were 17/17. Scheme 1 had the highest false positive rate [26.15‰ (11 075/423 524)] and the lowest positive predictive value [0.15% (17/11 092)]. Scheme 4 (Scheme 5) had the lowest false positive rate [0.02‰ (10/423 524)] and the highest positive predictive value [62.96% (17/27)]. Comparing scheme 4 (Scheme 5) with scheme 1, scheme 2 and scheme 3, the differences of false positive rate (χ2=302.30,11 191.50,32.06) and positive predictive value (χ2=102.51,3 485.61,13.83) were statistically significant (all P<0.001). Conclusion:C14∶1/C12∶1 was an effective auxiliary interpretive indicator for VLCADD in newborn screening, and the combination of C14∶1/C12∶1+C14∶1 was tested to be the best indicator for VLCADD screening based on non-derivatized tandem mass spectrometry.

Objective:To analyze the results of ATP7B gene screening in neonates and explore the linkage disequilibrium between different mutation loci, providing a basis for the clinical diagnosis and genetic counseling of Wilson′s disease.Methods:A total of 12 619 newborns who were born in Women′s Hospital of Nanjing Medical University during March 18 and December 30, 2022, including 6 605 male neonates and 6 014 female neonates, with birth weight of (3.44±0.56) kg, were retrospectively collected. The results of ATP7B gene screening in all newborns were analyzed.Next-generation sequencing technology was employed to detect the pathogenic loci of ATP7B gene, and the identified loci were verified using Sanger sequencing. PLINK 1.9 software was used to analyze the linkage disequilibrium of different mutation loci.Results:Among 12 619 neonates, 22 cases were diagnosed with 2-3 pathogenic mutations in the ATP7B gene (suspected positive). Among them, 20 cases were recalled for family verification, and 2 cases refused to recall. The verification results showed that 3 newborns had mutations of two loci respectively from their parents and were preliminarily diagnosed with Wilson′s disease, the other 17 neonates were carriers of the c.3316G>A/c.588C>A or c.1708-1G>C/c.1168A>G mutation loci arranged in a cis-acting manner from the father source or maternal source. A total of 249 pathogenic mutation carriers were detected (232 cases carrying 1 pathogenic mutation, and 17 cases carrying 2 pathogenic mutations), with a carrier rate of 1/51. Among them, the mutation c.2333G>T was most frequently detected (1/207), followed by c.2975C>T (1/421), c.2621C>T (1/742), c.2755C>G (1/971) and c.2605G>A (1/971). The results of linkage disequilibrium analysis in both c.3316G>A/c.588C>A and c.1708-1G>C/c.1168A>G showed that D ′=1, which showed complete linkage disequilibrium. Conclusion:The carrier rate of pathogenic mutations in the ATP7B gene is relatively high.Moreover, the c.3316G>A/c.588C>A and c.1708-1G>C/c.1168A>G pathogenic mutation loci are likely to be arranged in a cis-acting manner, highlighting the existence of linkage disequilibrium between the two groups of mutations. This finding provides important reference value for the clinical diagnosis and genetic counseling of Wilson disease.

The etiology and pathogenesis of depression is complex, the regulation of serotonin signaling pathway is one of the most important mechanisms for commercially available antidepressants. In addition to serotonin transporter, recent studies have shown many other serotonin receptor subtypes also play an important role in the treatment of depression, which has become one of the hot topics of antidepressants. Therefore, the progress of serotonin receptors and new structure with the activity of regulating serotonin receptors as well as part of the structure-activity relationships are reviewed in this paper in order to provide reference for the further research and development of antidepressants.

Objective To establish a discriminant method based on clinical and laboratory data and common examinations for early predicting the severity of pediatric infection. Methods Consecutive hospitalized patients diag-nosed as septic shock were included who were admitted between June 2014 and May 2015 retrospectively. Gender (male - female ratio:1. 25∶ 1. 00)and age(1 month to 6 years old)were matched in all of 18 patients with septic shock,and 27 patients diagnosed as systemic inflammatory response syndrome(SIRS),sepsis and severe sepsis on ad-mission were included respectively in order of sequential admission number during the same period. Additional 36 gen-der - and age - matched children with common infection(non - SIRS)were enrolled as controls. The clinical and labo-ratory examination data of all the included patients were collected and then the pediatric critical illness scores(PCIS) were made according to the worst condition within 24 hours of hospitalization. The parameters correlated with the severi-ty of infection were evaluated by rank correlation and Logistic regression analysis. The discriminant models were estab-lished based on κth - nearest - neighbor analysis and evaluated with clinical diagnosis by interrater agreement test. Results Except for platelet count,the other indexes including PCIS,neutrophil count,C - reactive protein,procalcito-nin(PCT),international normalized ratio of prothrombin time,activated partial thromboplastin time,thrombin time,fi-brinogen,fibrin/ fibrinogen degradation product(FDP)and D - dimer(D - D)all had differences among groups with varying infection severity(all P ﹤ 0. 001). The Spearman's coefficient ρ of PCIS,PCT,D - D and FDP correlated to in-fection severity were - 0. 837,0. 680,0. 679 and 0. 648,respectively(all P ﹤ 0. 001). Multivariate cumulative odds Lo-gistic regression analysis showed PCIS,D - D and PCT were related to infection severity(all P ﹤ 0. 05). The total error rate of discriminant models based on 3 - index combination(Mahalanobis transformation,k = 2)was 0. 091 that was lower than any models based on 2 - index combination or single - index. Using the discriminant model based on three -index combination,the infection severity of 26 patients admitted during June 2015 were predicted with a high interrater a-greement(weighted Kappa coefficient:0. 670,P ﹤ 0. 001)compared to clinical diagnosis. Conclusion The discriminant model based on combination of PCIS,D - D and PCT could assist predicting the severity of pediatric infection earlier.