[Clinical features and genetic analysis of five children with epilepsies due to variants of SCN8A gene].

OBJECTIVE: To explore the clinical and genetic characteristics of five children with epilepsies due to variants of SCN8A gene. METHODS: Clinical data of five children (four males and one female) admitted to Linyi People's Hospital due to hereditary epilepsies between August 2015 and August 2022 were collected. Whole exome sequencing was carried out for these children, and candidate variants were verified by Sanger sequencing. RESULTS: All of the five children were found to harbor variants of the SCN8A gene. Case 1, who had benign familial infantile epilepsy, inherited a known pathogenic c.4840A>G variant from his father with similar symptoms. Cases 2 to 4 had presented with intermediate epilepsy. Among these, case 2 has harbored a de novo c.3967G>A variant which was rated as pathogenic (PS1+PS2+PM1+PM2_Supporting+PP3) based on the guidelines from the American College of Medical Genetics and Genomics. Cases 3 and 4 were found to respectively harbor a de novo c.415A>T and a c.4697C>T variant, which were both rated as likely pathogenic (PS2+PM1+PM2_Supporting+PP3). Case 5, who had early-onset infantile epileptic encephalopathy transformed into Lennox Gastaut-like syndrome, has harbored a de novo c.5615G>A variant, which was known to be pathogenic. The children had their age of onset ranging from 2 to 14 months, and all had focal seizures and generalized tonic clonic seizures. Four children (cases 1, 2, 3 and 5) had cluster seizures, four (cases 1 to 4) had become seizure-free after single or dual treatment and showed normal growth and development, whilst case 5 was drug-resistant and showed severe developmental retardation. CONCLUSION: The five children had new features such as cluster seizures, occasional benign seizures in adulthood, and intermediate epilepsy which are prone to relapse after discontinuation of medication, which may be attributed to the pathogenic variants of the SCN8A gene.

[Genetic analysis of a family with epilepsy accompanied by developmental delay and brain deformity due to a de novo variant of TUBB2A gene].

OBJECTIVE: To explore the clinical manifestations and pathogenic variant in a family with epilepsy, developmental delay and brain deformity. METHODS: Clinical data of the child and his family members who had visited the Department of Pediatrics, Linyi People's Hospital on July 2, 2022 were collected. The child, his sister and parents were subjected to high-throughput sequencing, and the result was verified by Sanger sequencing. RESULTS: The child was a 6-year-old boy with developmentally delay and had epileptic seizures with fever sensitivity for four years. Cranial imaging showed brain dysplasia, while the video electroencephalogram showed abnormal discharge. High-throughput sequencing showed the child has harbored a heterozygous c.5G>T (p.Arg2Leu) variant of TUBB2A gene, which was unreported previously. His sister also carried the variant and had similar clinical manifestations, whilst his parents were of the wild-type and had normal clinical phenotypes. Based on the guidelines from the American College of Medical Genetics and Genomics (ACMG), the variant was classified as pathogenic (PS2+PM2_Supporting+PM5+PP1+PP2+PP3). CONCLUSION: The heterozygous c.5G>T (p.Arg2Leu) variant of the TUBB2A gene, in the form of gonadal mosaicism, probably underlay the disorders in this family.

Clinical phenotype and genetic characteristics of SZT2 related diseases: A case report and literature review.

PURPOSE: Seizure threshold 2 protein homolog gene (SZT2, MIM: 615463) related diseases are extremely rare autosomal recessive disorders with a wide spectrum of clinical phenotypes ranging from mild intellectual impairment to severe developmental epileptic encephalopathy (DEE). Most SZT2 related diseases are accompanied by craniofacial malformation and corpus callosum malformation. This study attempts to analyze and summarize the clinical phenotype and genetic characteristics of SZT2 related diseases, providing a basis for early diagnosis, treatment, and prognosis. METHOD: We analyzed the clinical characteristics of a Chinese child with pathogenic variants of SZT2. We also performed whole-exome sequencing (WES) on the patient. In addition, we conducted a literature review of previously reported patients with pathogenic mutations in the SZT2 gene. RESULT: The proband was a boy aged 1 year and 9 months with severe global developmental delay, transient drug-controlled focal epilepsy, cluster epilepsy, autism spectrum disorder, craniofacial deformity, hypotonia, focal EEG discharge, corpus callosum malformation, and persistent cavum septum pellucidum. WES revealed that the patient carried the SZT2 gene c.7584dupA and c.6302A>C complex heterozygous variants; the former being Likely Pathogenic (LP) and the latter Uncertain Significance (VUS) according to ACMG classification guidelines. According to our literature review, 43 cases of SZT2 related diseases have been reported so far; these include 15 cases with homozygous variations and 28 cases with complex heterozygous variations. A total of 57 types of variation were found, including 47 genetic variants, 2 de novo variants, and 8 unknown genetic modes. In addition, 2 high-frequency variants were found (c.5949_5951delTGT and c.6553C>T). The main clinical manifestations of the 40 patients were global developmental delay (GDD) of varying degrees (38/40, 95.00 %), seizures (36/40, 90.00 %), cranial deformity (27/40, 67.50 %), facial deformity (22/40, 55.00 %), hypotonia (22/40, 55.00 %), abnormal interseizure EEG discharge (26/40, 65.00 %), slow background activity (20/40, 50.00 %), corpus callosum deformity (18/40, 45.00 %). There was also one case of sudden unexpected death in epilepsy (SUDEP) and 3 cases of death from infection. In addition, three fetuses with the same variant had hydrocephalus and encephalocele. CONCLUSION: The compound heterozygous mutation of c.7584dupA and c.6302A>C in the SZT2 gene is the genetic etiology of this patient, expanding the mutation spectrum of SZT2 related diseases. Early genetic testing is the best choice for clear diagnosis, treatment, and prognosis.

[Genetic analysis and prenatal diagnosis of a child with Multiple congenital malformations-hypotonia-epilepsy syndrome type 3 due to variants of PIGT gene].

OBJECTIVE: To explore the clinical features and genetic etiology of a child with Multiple congenital malformations-hypotonia-epilepsy syndrome type 3 (MCAHS3) and provide prenatal diagnosis for her parents. METHODS: A female child who had presented at Linyi People's Hospital on 27 July 2022 for recurrent convulsions for over 4 years was selected as the study subject. Clinical data of the child were collected. Peripheral blood samples were taken from the child and her parents and subjected for whole exome sequencing (WES). Candidate variants were verified by Sanger sequencing. Prenatal diagnosis was carried out on amniotic fluid sample at 18 weeks' gestation. Bioinformatic software was used to analyze the pathogenicity of the protein model for the variant loci. RESULTS: The child was a 4-year-old female with frequent seizures, peculiar facial appearance, hypotonia and severe developmental delay. Genetic analysis revealed that she has harbored compound heterozygous variants of the PIGT gene, namely c.1126del (p.H376Tfs*56) and c.1285G>C (p.E429Q), which were respectively inherited from her mother and father. Based on the guidelines from the American College of Medical Genetics and Genomics, the c.1126del (p.H376Tfs*56) variant was predicted to be pathogenic (PVS1+PM2_Supporting+PM4), and c.1285G>C (p.E429Q) variant was predicted to be likely pathogenic (PM2_Supporting+PM3+PM4). Prenatal diagnosis suggested that the fetus also harbored the same compound heterozygous variants, and the pregnancy was terminated with induced labor. CONCLUSION: The c.1126del (p.H376Tfs*56) and c.1285G>C (p.E429Q) compound heterozygous variants of the PIGT gene probably underlay the MCAHS3 in this patient, and prenatal diagnosis has prevented birth of further affected child in this family.

A novel heterozygous ATP1A2 pathogenic variant in a Chinese child with MELAS-like alternating hemiplegia.

BACKGROUND: Pathogenic variants of ATP1A2 (OMIM ID: 182340) are usually associated with familial hemiplegic migraine type 2 (FHM-2), alternating hemiplegia of childhood (AHC), early infantile epileptic encephalopathy (EIEE), transient cytotoxic edema, and so on. Here, we present a novel heterozygous ATP1A2 variant in a girl with alternating hemiplegia, febrile seizures, developmental delay (which subsequently subsided), and MELAS-like syndrome (as indicated by brain MRI). The patient did not experience migraine with aura. METHODS: The patient was an 8-year-old girl with normal growth and development. Beginning from the age of 3 years and 8 months, the patient experienced several episodes of alternating limb paralysis. The episodes were accompanied by the appearance of MELAS-like findings on brain MRI, which corresponded to the hemiplegia. There were abnormal linear signals in the cerebral cortex on the opposite side of the hemiplegic limb. Each time the patient recovered from hemiplegia, and each time MRI showed no lesions remained after recovery. No obvious abnormality was found in other examinations. Finally, the patient underwent whole-exome sequencing (WES). RESULTS: WES revealed a novel and de novo heterozygous variant in the ATP1A2 (NM_000702.3) c.335C>A:p.Ala112Asp (not previously reported). We examined the variant position in the 3D protein structure and found that a missense mutation at this site is a nonconservative substitution. The variation is nonpolymorphic. It occurs at a very low frequency in the population, and its ACMG classification is likely pathogenic. CONCLUSION: At present, there are limited reports of mutations in the ATP1A2 gene causing AHC. This is the first case of brain MRI showing MELAS-like imaging in an AHC patient, and more cases are needed for verification. Early genetic testing and family screening can aid in the diagnosis and treatment of genetic diseases. The relationship between ATP1A2 gene mutation genotype and clinical phenotype needs to be further studied.

A novel composition of two heterozygous GFM1 mutations in a Chinese child with epilepsy and mental retardation.

INTRODUCTION: G elongation factor mitochondrial 1 (GFM1) encodes one of the mitochondrial translation elongation factors. GFM1 variants were reported to be associated with neurological diseases and liver diseases in a few cases. Here, we present a novel composition of two heterozygous mutations of GFM1 in a boy with epilepsy, mental retardation, and other unusual phenotypes. METHODS: The patient was found to be blind and experienced recurrent convulsive seizures such as nodding and hugging at the age of 3 months. After antiepileptic treatment with topiramate, he had no obvious seizures but still had mental retardation. The patient vomited frequently at 16 months old, sometimes accompanied by epileptic seizures. Hematuria metabolic screening, mutation screening of mitochondrial gene, and mitochondrial nuclear gene were negative. Then, he was analyzed by whole-exome sequencing (WES). RESULTS: Whole-exome sequencing revealed a novel composition of two heterozygous mutations in GFM1, the maternal c.679G > A (has not been reported) and the paternal c.1765-1_1765-2del (previously reported). At present, there is no specific and effective treatment for the disease, and the prognosis is very poor. CONCLUSION: The discovery of new phenotypes and new genotypes will further enrich the diagnosis information of the disease and provide more experiences for clinicians to quickly diagnose the disease and judge the prognosis.

Novel and de novo point and large microdeletion mutation in PRRT2-related epilepsy.

BACKGROUND: Point and copy number variant mutations in the PRRT2 gene have been identified in a variety of paroxysmal disorders and different types of epilepsy. In this study, we analyzed the phenotypes and PRRT2-related mutations in Chinese epilepsy children. METHODS: A total of 492 children with epilepsy were analyzed by whole exome sequencing (WES) and low-coverage massively parallel CNV sequencing (CNV-seq) to find the single nucleotide variants and copy number variations (CNVs). And quantitative polymerase chain reaction was utilized to verify the CNVs. Their clinical information was followed up. RESULTS: We found PRRT2-related mutations in 19 patients (10 males and nine females, six sporadic cases and 13 family cases). Twelve point mutations, four whole gene deletion, and three 16p11.2 deletions were detected. The clinical features of 39 patients in 19 families included one early childhood myoclonic epilepsy (ECME), one febrile seizure (FS), two infantile convulsions with paroxysmal choreoathetosis (ICCA), six paroxysmal kinesigenic dyskinesias (PKD), 12 benign infantile epilepsy (BIE), and 17 benign familial infantile epilepsy (BFIE). All patients had normal brain MRI. Interictal EEG showed only one patient had generalized polyspike wave and five patients had focal transient discharges. Focal seizures originating in the frontal region were recorded in one patient, two from the temporal region, and two from the occipital region. Most patients were treated effectively with VPA or OXC, and the child with myoclonic seizures was not sensitive to antiepileptic drugs. CONCLUSION: PRRT2 mutations can be inherited or de novo, mainly inherited. The clinical spectrum of PRRT2 mutation includes BIE, BFIE, ICCA, PKD, FS, and ECME. The PRRT2-related mutations contained point mutation, whole gene deletion and 16p11.2 deletions, and large microdeletion mutations mostly de novo. It is the first report of PRRT2 mutation found in ECME. Our report expands the mutation and clinical spectrum of PRRT2-related epilepsy.

Conventional vs. pulsed-light accelerated corneal collagen cross-linking for the treatment of progressive keratoconus: 12-month results from a prospective study.

The aim of the present study was to compare the clinical outcomes of conventional corneal collagen cross-linking (CXL) and pulsed-light accelerated CXL (pl-ACXL) in the eyes of patients with progressive keratoconus. A total of 72 eyes with progressive keratoconus in 58 patients were equally divided into the CXL and pl-ACXL treatment groups. The CXL treatment was performed using the UVX 1000 system with 0.1% riboflavin solution in 20% dextran presoak for 30 min, and 3 mW/cm2 ultraviolet A (UVA) light for 30 min. The pl-ACXL group was treated with the KXL system using 0.1% riboflavin with HPMC presoak for 10 min, followed by 8 min (1 sec on/1 sec off) of 30 mW/cm2 UVA light. Patients were evaluated according to the uncorrected distance visual acuity (UDVA), corrected DVA (CDVA), refraction, maximum keratometry (Kmax), endothelial cell density (ECD), anterior segment optical coherence tomography and in vivo confocal microscopy. The follow-up period was 12 months. Transient haze was observed in 17 eyes (47.22%) in the CXL group and 8 eyes (22.22%) in the pl-ACXL group at 1 month postoperatively. There were no significant postoperative differences in the astigmatism, manifest refraction spherical equivalent, ECD or thinnest corneal thickness. By contrast, UDVA, CDVA and Kmax presented significant improvement at 12 months postoperatively in the two groups. The demarcation line depth was 284.94±33.29 µm in the CXL group, which was significantly deeper in comparison with that in the pl-ACXL group (201.64±27.72 µm; P<0.01) at 1 month postoperatively. In vivo confocal microscopy revealed keratocyte apoptosis and stromal edema at 1 month postoperatively, which gradually recovered towards the normal status after 12 months in the two groups. There were no apparent changes in the posterior stroma and endothelium in either group. The results of the present study revealed that CXL and pl-ACXL were safe and effective procedures in stabilizing the progression of keratoconus. The CXL technique offers more effective visual and topographic outcomes compared with pl-ACXL, while pl-ACXL ensures shorter treatment time and reduced microstructural damage.

Lack of association between SLC5A7 polymorphisms and Tourette syndrome in a Chinese Han population.

Although Tourette syndrome (TS) is a chronic neuropsychiatric disorder whose pathogenesis remains unclear, genetic factors play an important role in the occurrence and development. A variety of studies have been shown that the candidate genes related to cholinergic neurons may be associated with the onset of TS. To investigate the association between the SLC5A7 polymorphisms and Tourette syndrome (TS) in the Chinese Han population, the SNP rs1013940, rs2433718, and rs4676169 were genotyped in 401 TS trios and 400 controls. The transmission disequilibrium test (TDT) and haplotype relative risk (HRR) compared genetic distributions of trios, while the chi-square test compared patients and controls. However, no transmission disequilibrium was found between the three SLC5A7 SNPs and TS. Therefore, we think that this gene may not be the main risk factor on the onset of TS. However, these results should be further validated in different populations.

Therapeutic and inducing effect of corneal crosslinking on infectious keratitis.

The corneal crosslinking (CXL) with riboflavin and ultraviolet-A (UVA) is a new therapy method to successfully treat infectious keratitis in clinical practice. However, there are rare reports on the complications of CXL such as the secondary keratitis. The diverse clinical outcomes on keratitis have highlighted the necessity to further evaluate the efficacy and complications of CXL. We reviewed the positive and negative reports on UVA/riboflavin related with keratitis and provided our opinion on the therapeutic and side effect of UVA/riboflavin crosslinking on keratitis.

[Effects of macrophage migration inhibitory factor in pathogenesis of acute respiratory distress syndrome in children].

OBJECTIVE: Acute respiratory distress syndrome (ARDS) is usually diagnosed on the basis of clinical manifestations. However, a sensitive and effective biochemical index is important for early diagnosis of ARDS. It has been confirmed that macrophage migration inhibitory factor (MIF) expression is increased in patients with ARDS (adults and children). The present study aimed to investigate the expression and pathogenic role of MIF in children with ARDS by determining the serum level of MIF and expression of MIF in lung tissues. METHODS: Totally 18 children with ARDS, who were diagnosed in the department of pediatrics, the People's Hospital of Nanhai District, and 8 healthy children (control) were enrolled into the study. The serum level of MIF in ARDS children and normal children were measured by using enzyme-linked immunosorbent assay (ELISA). Peripheral blood mononuclear cell (PBMC) MIF expression was determined by flow-cytometry. The expression of MIF mRNA and protein in lung tissues were detected by using double-staining immunohistochemistry and in situ hybridization. RESULTS: (1) The serum level of MIF and PBMC MIF expression were (2040 +/- 146) microg/L and (8.98 +/- 2.76)%, respectively in ARDS children, which were significantly higher than those in the healthy children [(53 +/- 31) microg/L and (0.97 +/- 0.28)%, (P < 0.01)]. (2) In situ hybridization and immunohistochemistry showed that the number of KP(1)(+) cells and the percentages of MIF(+) expressed cells and MIF mRNA expressed cells were (229 +/- 87)/mm(2), (31.4 +/- 7.8)% and (34.71 +/- 8.91)%, respectively in the bronchoalveolar lavage fluid of ARDS children, which were significantly higher than those in the healthy children [(11 +/- 6)/mm(2), (1.9 +/- 0.8)% and (1.17 +/- 0.16)%, P < 0.01)]. (3) The number of KP(1)(+) cells and the percentages of MIF(+) expressed cells and MIF mRNA expressed cells were (319 +/- 129)/mm(2), (41.7 +/- 11.6)% and (45.13 +/- 13.2)% in ARDS lungs interstitium, which were significantly higher than those in the healthy children [(11 +/- 6)/mm(2), (1.9 +/- 0.8)% and (1.40 +/- 0.25)%, (P < 0.01)]. CONCLUSIONS: The level of MIF increased in serum and lung interstitium in ARDS children. Macrophage infiltration was demonstrated in lung interstitium and bronchoalveolar lavage fluid of ARDS children, MIF was expressed and significantly associated with the number of macrophage, which suggest that MIF plays an important role in the pathogenesis of ARDS. The level of MIF expression and macrophage infiltration can be regarded as a sensitive and effective biochemical index in the early diagnosis of ARDS.