A c.726C>G (p.Tyr242Ter) nonsense mutation-associated with splicing alteration (NASA) of WDR45 gene underlies ß-propeller protein-associated neurodegeneration (BPAN).

Neurodegeneration with brain iron accumulation (NBIA) is a clinically and genetically heterogeneous disease characterized by increased iron deposition in the basal ganglia and progressive degeneration of the nervous system in adulthood. However, in early childhood, there were no characteristic features to perform early diagnosis. In our study, a female child exhibited global developmental delay, intellectual disability, and febrile seizure without other distinct clinical phenotypes. Through whole exome sequencing (WES), a de novo nonsense mutation (c.726C > G, p. Tyr242Ter) of WDR45 gene was identified in this child. She was finally diagnosed as ß-propeller protein-associated neurodegeneration (BPAN), one of the recently identified subtypes of NBIA. This mutation could act as a premature stop codon (PSC) which rendered the mutated transcripts to be degraded by nonsense-mediated mRNA decay (NMD), leading to decreased levels of PSC-containing mRNAs. Additionally, through mini-gene splicing assays, this mutation could result in an unprecedented novel transcript with the exon 9 of WDR45 excluded by nonsense-associated splicing alteration (NASA). Transcriptome sequencing (RNA-seq) on total RNAs from PBMCs of the trio revealed three types of alternative splicing events in the patient. Further research implied that downregulation of iron transport genes (TFRC, TFR2, SCARA5) might be the underlying mechanism for the iron accumulation in patients with deficient WDR45. This is the first report about NASA happening in WDR45. It implies that nonsense mutations approximal to splicing sites could affect the disease pathogenesis through more than one molecular mechanism and should be taken into consideration when conducting genetic counseling.

[Differential diagnosis of autism spectrum disorder and global developmental delay based on machine learning and Children Neuropsychological and Behavioral Scale]. / åŸºäºŽæœºå™¨å­¦ä¹ å’Œå„¿ç«¥ç¥žç»å¿ƒç†è¡Œä¸ºæ£€æŸ¥é‡è¡¨é‰´åˆ«å­¤ç‹¬ç—‡è°±ç³»éšœç¢å’Œå ¨é¢å‘è‚²è¿Ÿç¼“å„¿ç«¥çš„ç ”ç©¶.

OBJECTIVES: To investigate the efficacy and required indicators of Children Neuropsychological and Behavioral Scale-Revision 2016 (CNBS-R2016) in the differential diagnosis of autism spectrum disorder (ASD) and global developmental delay (GDD). METHODS: A total of 277 children with ASD and 415 children with GDD, aged 18-48 months, were enrolled as subjects. CNBS-R2016 was used to assess the developmental levels of six domains, i.e., gross motor, fine motor, adaptive ability, language, social behavior, and warning behavior, and a total of 13 indicators on intelligence age and developmental quotient (DQ) were obtained as the input features. Five commonly used machine learning classifiers were used for training to calculate the classification accuracy, sensitivity, and specificity of each classifier. RESULTS: DQ of warning behavior was selected as the first feature in all five classifiers, and the use of this indicator alone had a classification accuracy of 78.90%. When the DQ of warning behavior was used in combination with the intelligence age of warning behavior, gross motor, and language, it had the highest classification accuracy of 86.71%. CONCLUSIONS: Machine learning combined with CNBS-R2016 can effectively distinguish children with ASD from those with GDD. The DQ of warning behavior plays an important role in machine learning, and its combination with other features can improve classification accuracy, providing a basis for the efficient and accurate differential diagnosis of ASD and GDD in clinical practice.

Identification of a de novo mutation of the FOXG1 gene and comprehensive analysis for molecular factors in Chinese FOXG1-related encephalopathies.

Background: FOXG1-related encephalopathy, also known as FOXG1 syndrome or FOXG1-related disorder, affects most aspects of development and causes microcephaly and brain malformations. This syndrome was previously considered to be the congenital variant of Rett syndrome. The abnormal function or expression of FOXG1, caused by intragenic mutations, microdeletions or microduplications, was considered to be crucial pathological factor for this disorder. Currently, most of the FOXG1-related encephalopathies have been identified in Europeans and North Americans, and relatively few Chinese cases were reported. Methods: Array-Comparative Genomic Hybridization (Array-CGH) and whole-exome sequencing (WES) were carried out for the proband and her parent to detect pathogenic variants. Results: A de novo nonsense mutation (c.385G>T, p.Glu129Ter) of FOXG1 was identified in a female child in a cohort of 73 Chinese children with neurodevelopmental disorders/intellectual disorders (NDDs/IDs). In order to have a comprehensive view of FOXG1-related encephalopathy in China, relevant published reports were browsed and twelve cases with mutations in FOXG1 or copy number variants (CNVs) involving FOXG1 gene were involved in the analysis eventually. Feeding difficulties, seizures, delayed speech, corpus callosum hypoplasia and underdevelopment of frontal and temporal lobes occurred in almost all cases. Out of the 12 cases, eight patients (66.67%) had single-nucleotide mutations of FOXG1 gene and four patients (33.33%) had CNVs involving FOXG1 (3 microdeletions and 1 microduplication). The expression of FOXG1 could also be potentially disturbed by deletions of several brain-active regulatory elements located in intergenic FOXG1-PRKD1 region. Further analysis indicated that PRKD1 might be a cooperating factor to regulate the expression of FOXG1, MECP2 and CDKL5 to contribute the RTT/RTT-like disorders. Discussion: This re-analysis would broaden the existed knowledge about the molecular etiology and be helpful for diagnosis, treatment, and gene therapy of FOXG1-related disorders in the future.

A synonymous variant contributes to a rare Wiedemann-Rautenstrauch syndrome complicated with mild anemia via affecting pre-mRNA splicing.

Wiedemann-Rautenstrauch syndrome (WDRTS) is an extremely rare autosomal recessive neonatal disorder. Currently, over 50 cases with variable phenotypes of WDRTS have been reported. In our cohort of prenatal and postnatal growth retardation, a female proband was found to have general growth retardation, neurocutaneous syndrome, and anemia. Karyotype test and array-CGH detected no obvious chromosomal aberrations. Trio-based whole-exome sequencing (Trio-WES) identified bi-allelic compound mutations in the coding sequence (CDS) of POLR3A gene (c.3342C > T, p.Ser1114 = and c.3718G > A, p.Gly1240Ser). For the mild anemia phenotype, the underlying causal genetic factors could be attributed to the compound heterozygous mutations in FANCA gene (c.2832dup, p.Ala945CysfsTer6 and c.1902 T > G, p.Asp634Glu). Mini-gene reporter assays revealed that the synonymous variant of POLR3A and the missense variant of FANCA could affect pre-mRNA splicing of each gene. For POLR3A, the synonymous mutation (c.3342C > T, p.Ser1114=) generated three types of aberrant isoforms. Therefore, the female patient was finally diagnosed as WDRTS caused by POLR3A. For FANCA, the missense variant (c.1902 T > G, p.Asp634Glu) disrupted the normal splicing between exon 21 and 22, and produced two types of abnormal isoforms, one carrying the 1902G and the other spliced between exon 21 and 23 to exclude exon 22. Network analysis showed that POLR3A and FANCA could be STRINGed, indicating both proteins might collaborate for some unknown functions. Current investigation would broaden the knowledge for clinicians and genetic counselors and remind them to interpret those synonymous or predicted "benign" variants more carefully.

A de Novo ZMIZ1 Pathogenic Variant for Neurodevelopmental Disorder With Dysmorphic Facies and Distal Skeletal Anomalies.

Background: Neurodevelopmental disorder with dysmorphic facies and distal skeletal anomalies (NEDDFSA) is a rare syndromic disorder characterized by global neurodevelopmental delay, early-onset hypotonia, poor overall growth, poor speech/language ability, and additional common phenotypes such as eye anomalies, joint hypermobility, and skeletal anomalies of the hands and feet. NEDDFSA is caused by heterozygous pathogenic variants in the ZMIZ1 gene on chromosome 10q22.3 with autosomal dominant (AD) mode of inheritance. All the 32 reported cases with variants in ZMIZ1 gene had a genetic background in Caucasian, Hispanic, North African, and Southeastern Asian. Until now, there are no reports of Chinese patients with ZMIZ1 pathogenic variants. Methods: A 5-year-old girl was found to have the characteristic phenotypes of NEDDFSA. Array-Comparative Genomic Hybridization (array-CGH) and whole exome sequencing (WES) were applied for the trio of this female patient. Sanger sequencing was used to verify the selected variants. A comprehensive molecular analysis was carried out by protein structure prediction, evolutionary conservation, motif scanning, tissue-specific expression, and protein interaction network to elucidate pathogenicity of the identified ZMIZ1 variants. Results: The karyotype was 46, XX with no micro-chromosomal abnormalities identified by array-CGH. There were 20 variants detected in the female patient by WES. A de novo heterozygous missense variant (c.2330G > A, p.Gly777Glu, G777E) was identified in the exon 20 of ZMIZ1. No variants of ZMIZ1 were identified in the non-consanguineous parents and her healthy elder sister. It was predicted that G777E was pathogenic and detrimental to the spatial conformation of the MIZ/SP-RING zinc finger domain of ZMIZ1. Conclusion: Thus far, only four scientific articles reported deleterious variants in ZMIZ1 and most of the cases were from Western countries. This is the first report about a Chinese patient with ZMIZ1 variant. It will broaden the current knowledge of ZMIZ1 variants and variable clinical presentations for clinicians and genetic counselors.

Assessment of growth pattern of preterm infants up to a corrected age of 24 months. / æ—©äº§å„¿æ ¡æ­£24æœˆé¾„å† ç”Ÿé•¿è½¨è¿¹ç ”ç©¶.

OBJECTIVES: To assess the growth of preterm infants up to a corrected age of 24 months, and to understand the growth trend and pattern of preterm infants. METHODS: A preterm infant follow-up database was established based on the Internet Plus follow-up system. A total of 3 188 preterm infants who were born from April 2018 to April 2021 were enrolled. Their length, weight, and head circumference were recorded at birth and at the corrected ages of 1, 3, 6, 12, 18, and 24 months. The preterm infants were grouped by perinatal factors. The growth curves of these infants were plotted and compared with the International Fetal and Newborn Growth Consortium for the 21st Century (INTERGROWTH-21st) standard and World Health Organization (WHO) standard. RESULTS: The weight, length, and head circumference curves of each group of preterm infants grouped by various perinatal factors all rose rapidly within the corrected age of 6 months, but the growth rate slowed down after the corrected age of 6 months. Based on the actual age for the groups of preterm infants with different gestational ages (<28 weeks, 28-31+6 weeks, 32-33+6 weeks, and 34-36+6 weeks), the length curve gradually coincided with the WHO curve after the actual age of 9 months (P=0.082), while for the preterm infants with a gestational age of <32 weeks, the weight and head circumference curves were significantly lower than the WHO curves (P<0.001). Based on the corrected age, the physical growth curve of preterm infants with different gestational ages (<28 weeks, 28-31+6 weeks, 32-33+6 weeks, and 34-36+6 weeks) basically coincided with each other (P>0.05). For the infants with extremely low birth weight and the small-for-gestational-age infants, the length, weight, and head circumference curves were significantly lower than those of the INTERGROWTH-21st standard and the WHO standard (P<0.05). CONCLUSIONS: The physical growth rate of preterm infants is faster within the corrected age of 6 months, and the growth rate slows down after the corrected age of 6 months. Preterm infants with a smaller gestational age need longer time to catch up in weight and head circumference. More attention should be paid to the physical growth of extremely preterm infants, extremely low birth weight infants, and small-for-gestational-age infants.

Restricted Kinematics in Children With Autism in the Execution of Complex Oscillatory Arm Movements.

Restricted and repetitive behavior is a core symptom of autism spectrum disorder (ASD) characterized by features of restrictedness, repetition, rigidity, and invariance. Few studies have investigated how restrictedness is manifested in motor behavior. This study aimed to address this question by instructing participants to perform the utmost complex movement. Twenty children with ASD and 23 children with typical development (TD) performed one-dimensional, left-right arm oscillations by demonstrating varying amplitudes and frequencies. The entropy of amplitude and velocity was calculated as an index of kinematic complexity. Results showed that the velocity entropy, but not the amplitude entropy, was significantly lower in ASD than in TD (p < 0.01), suggesting restricted kinematics. Further analysis demonstrated that a significantly higher proportion of the velocity values was allocated at a low-speed level in the children with ASD (p < 0.01). A qualitative comparison of the complex movement with movement at preferred frequency suggested that the children with ASD might be less likely to shift away from the preferred movement. However, our study can be improved in terms of recruiting a larger sample of participants, measuring the level of motivation, and collecting both complex and preferred movements of the same participant.

Identification of de novo mutations for ARID1B haploinsufficiency associated with Coffin-Siris syndrome 1 in three Chinese families via array-CGH and whole exome sequencing.

BACKGROUND: Coffin-Siris syndrome (CSS) is a multiple malformation syndrome characterized by intellectual disability associated with coarse facial features, hirsutism, sparse scalp hair, and hypoplastic or absent fifth fingernails or toenails. CSS represents a small group of intellectual disability, and could be caused by at least twelve genes. The genetic background is quite heterogenous, making it difficult for clinicians and genetic consultors to pinpoint the exact disease types. METHODS: Array-Comparative Genomic Hybridization (array-CGH) and whole exome sequencing (WES) were applied for three trios affected with intellectual disability and clinical features similar with those of Coffin-Siris syndrome. Sanger sequencing was used to verify the detected single-nucleotide variants (SNVs). RESULTS: All of the three cases were female with normal karyotypes of 46, XX, born of healthy, non-consanguineous parents. A 6q25 microdeletion (arr[hg19]6q25.3(155,966,487-158,803,979) × 1) (2.84 Mb) (case 1) and two loss-of-function (LoF) mutations of ARID1B [c.2332 + 1G > A in case 2 and c.4741C > T (p.Q1581X) in case 3] were identified. All of the three pathogenic abnormalities were de novo, not inherited from their parents. After comparison of publicly available microdeletions containing ARID1B, four types of microdeletions leading to insufficient production of ARID1B were identified, namely deletions covering the whole region of ARID1B, deletions covering the promoter region, deletions covering the termination region or deletions covering enhancer regions. CONCLUSION: Here we identified de novo ARID1B mutations in three Chinese trios. Four types of microdeletions covering ARID1B were identified. This study broadens current knowledge of ARID1B mutations for clinicians and genetic consultors.

Late-onset sepsis in very low birth weight preterm infants: 7 years' experience at a tertiary hospital in China.

OBJECTIVE: Studies about the epidemiology and risk factors of late onset of sepsis (LOS) in preterm very low birth weight (PVLBW) infants of developing countries are limited and variable. The objective of this study was to explore the epidemiology, risk factors and pathogens of LOS in VLBW infants in a Chinese tertiary hospital. METHODS: Data concerning demographies, pathogens, risk factors and outcomes of LOS were collected in a cohort of 710 VLBW infants discharged from January 2012 to December 2018. Chi-square, t-test and multivariable logistic regression analysis were used to identify risk factors. RESULTS: The overall incidence of LOS was 24.08% and blood culture-proven sepsis was 19.58%. Gram-negative bacteria accounted for 57.93%, and 32.41% of the isolates were Klebsiella pneumoniae. LOS led to a longer hospital stay, higher hospitalization cost and higher mortality compared to infants without LOS. Mortality in Gram-negative or fungi LOS was higher than in Gram-positive LOS. The factor of PVLBW infants with registered permanent residence (PR) was associated with 80% lower likelihood of LOS compared to those without registered PR (P = 0.000, 95% CI 0.096-0.420). Infants with thrombocytopenia had 2.5 times LOS of control (P = 0.014, CI 1.210-5.132). Duration of parenteral nutrition was a risk of LOS (P = 0.009, CI 1.010-1.069). CONCLUSION: Gram-negative bacteria, especially Klebsiella pneumonia, were the dominant pathogens in this study. Social factors might be a risk of LOS in PVLBW infants in developing countries. Taking measures to reduce the duration of parenteral nutrition may be helpful in decreasing the incidence of LOS. Awareness of thrombocytopenia may help to recognized diagnosis of LOS.

Atypical Head Movement during Face-to-Face Interaction in Children with Autism Spectrum Disorder.

The present study implemented an objective head pose tracking technique-OpenFace 2.0 to quantify the three dimensional head movement. Children with autism spectrum disorder (ASD) and typical development (TD) were engaged in a structured conversation with an interlocutress while wearing an eye tracker. We computed the head movement stereotypy with multiscale entropy analysis. In addition, the head rotation range (RR) and the amount of rotation per minute (ARPM) were calculated to quantify the extent of head movement. Results demonstrated that the ASD group had significantly higher level of movement stereotypy, RR and ARPM in all the three directions of head movement. Further analyses revealed that the extent of head movement could be significantly explained by movement stereotypy, but not by the amount of visual fixation to the interlocutress. These results demonstrated the atypical head movement dynamics in children with ASD during live interaction. It is proposed that head movement might potentially provide novel objective biomarkers of ASD. LAY SUMMARY: Our study used an objective tool to quantify head movement in children with autism. Results showed that children with autism had more stereotyped and greater head movement. We suggest that head movement tracking technique be widely used in autism research.

[Application of the Children Neuropsychological and Behavioral Scale-Revision 2016 in young children with autism spectrum disorder].

OBJECTIVE: To compare the assessment results of the Children Neuropsychological and Behavioral Scale-Revision 2016 (CNBS-R2016) between young children with autism spectrum disorder (ASD) and global developmental delay (GDD, without ASD) and to explore whether CNBS-R2016 could be helpful to early identification of ASD. METHODS: A total of 260 ASD and 371 GDD children aged 18-30 months were enrolled to finish the assessment of CNBS-R2016. The development quotients (DQs) of the five domains of CNBS-R2016 including gross motor, fine motor, adaptability, language and social behavior were compared between the two groups. The receiver operating characteristic (ROC) curve was used to evaluate the value of the autism-predicted domain in identifying ASD and GDD. RESULTS: The DQs of all the five domains in the ASD group were lower than those in the GDD group (P<0.05). The language DQ and total DQ of the ASD group had a negative correlation with the score of the autism-predicted domain (rs=-0.566, -0.552 respectively, P<0.01). When the cut-off value of the autism-predicted domain was 10.5, the largest area under the ROC curve was 0.835, and the sensitivity and specificity for the diagnosis of ASD were 0.750 and 0.798 respectively. CONCLUSIONS: The development of ASD children aged 18-30 months is worse than that of GDD children. CNBS-R2016 may be helpful to distinguish ASD from children with developmental delay.

Altered composition and function of intestinal microbiota in autism spectrum disorders: a systematic review.

At present, the pathophysiology of autism spectrum disorder (ASD) remains unclear. Increasing evidence suggested that gut microbiota plays a critical role in gastrointestinal symptoms and behavioral impairment in ASD patients. The primary aim of this systematic review is to investigate potential evidence for the characteristic dysbiosis of gut microbiota in ASD patients compared with healthy controls (HCs). The MEDLINE, EMBASE, Web of Science and Scopus were systematically searched before March 2018. Human studies that compared the composition of gut microbiota in ASD patients and HCs using culture-independent techniques were included. Independent data extraction and quality assessment of studies were conducted according to PRISMA statement and Newcastle-Ottawa Scale. Phylogenetic Investigation of Communities by Reconstruction of Unobserved States (PICRUSt) was used to infer biological functional changes of the shifted microbiota with the available data in four studies. Sixteen studies with a total sample size of 381 ASD patients and 283 HCs were included in this systematic review. The quality of the studies was evaluated as medium to high. The overall changing of gut bacterial community in terms of ß-diversity was consistently observed in ASD patients compared with HCs. Furthermore, Bifidobacterium, Blautia, Dialister, Prevotella, Veillonella, and Turicibacter were consistently decreased, while Lactobacillus, Bacteroides, Desulfovibrio, and Clostridium were increased in patients with ASD relative to HCs in certain studies. This systematic review demonstrated significant alterations of gut microbiota in ASD patients compared with HCs, strengthen the evidence that dysbiosis of gut microbiota may correlate with behavioral abnormality in ASD patients. However, results of inconsistent changing also existed and further big-sampled well-designed studies are needed. Generally, as a potential mediator of risk factors, the gut microbiota could be a novel target for ASD patients in the future.

Elevated levels of cerebrospinal fluid S100B are associated with brain injury and unfavorable outcomes in children with central nervous system infections.

PURPOSE: This work aimed to assess whether elevated levels of cerebrospinal fluid (CSF) S100B are associated with brain injury and unfavorable outcomes at discharge in children with central nervous system (CNS) infections. METHODS: CSF S100B and associated clinical parameters were retrospectively analyzed in 83 children with CNS infections and 88 children without neurological pathology served as controls. Children with CNS infections were divided into an infectious encephalitis group and an infectious meningitis group based on whether cerebral parenchyma was involved, and CSF S100B levels in different age subgroups between the two groups were compared. The predictive value of CSF S100B in children with infectious encephalitis was evaluated by multivariate logistic regression analysis, and the discriminative power was investigated by receiver operating characteristic (ROC) analysis. RESULTS: CSF S100B levels in the infectious encephalitis group were significantly higher than the infectious meningitis and the control group at each age range. CSF S100B ≥ 0.96 µg/L had 62.9% sensitivity and 76.2% specificity for diagnosing cerebral parenchyma injury in children with CNS infections. Increased CSF S100B levels were proven to be an independent predictor of unfavorable outcomes in children with infectious encephalitis and the optimal cut-off value (1.77 µg/L of CSF S100B) for predicting unfavorable outcomes in children with infectious encephalitis showed 61.1% sensitivity and 96.2% specificity. CONCLUSIONS: This study has demonstrated that elevated levels of CSF S100B are associated with brain injury and could be used as an independent predictor of clinically unfavorable outcomes at discharge in children with CNS infections.

Effects of hyperoxia on the dynamic expression of Aquaporin5 in premature rats lung development.

To explore the dynamic expression and role of Aquaporin5 (AQP5) in lung development and hyperoxia lung injury, gestation 21-day Sprague-Dawley (SD) rats (term=22 days) were randomly assigned to air group and hyperoxia group within 12-24 h after birth. The rats in hyperoxia group were continuously exposed to about 85% oxygen and those in air group to room air. After 1 to 14 days of exposure, total lung RNA was extracted and the expression of AQP5 mRNA was detected by reverse transcription polymerase chain reaction (RT-PCR). Immunohistochemistry and western-blot were used to detect the expression of AQP5 protein. The results showed that the expression of AQP5 in premature rats lung could be detected at various time points after birth, and the positive staining was restricted to the type I alveolar epithelial cells. In air group, the AQP5 expression was detected in a very low level at day 1, but exhibited a persistent increase after birth. Compared with the air group, the expression of AQP5 in hyperoxia group was increased at day 1, and had significant difference in mRNA level (P<0.05), but decreased significantly in mRNA and protein levels after 4 to 14 days (P<0.01 or P<0.05 respectively). It was concluded that AQP5 might play a key role in the alveolar period of premature rats by regulating the lung water balance. Hyperoxia exposure leads to a down-regulation of the AQP5 expression, which may be an important factor for the development of hyperoxia lung injury.