Association of FAT1 with focal epilepsy and correlation between seizure relapse and gene expression stage.

PURPOSE: The FAT1 gene encodes FAT atypical cadherin 1, which is essential for foetal development, including brain development. This study aimed to investigate the relationship between FAT1 variants and epilepsy. METHODS: Trio-based whole-exome sequencing was performed on a cohort of 313 patients with epilepsy. Additional cases with FAT1 variants were collected from the China Epilepsy Gene V.1.0 Matching Platform. RESULTS: Four pairs of compound heterozygous missense FAT1 variants were identified in four unrelated patients with partial (focal) epilepsy and/or febrile seizures, but without intellectual disability/developmental abnormalities. These variants presented no/very low frequencies in the gnomAD database, and the aggregate frequencies in this cohort were significantly higher than those in controls. Two additional compound heterozygous missense variants were identified in two unrelated cases using the gene-matching platform. All patients experienced infrequent (yearly/monthly) complex partial seizures or secondary generalised tonic-clonic seizures. They responded well toantiseizure medication, but seizures relapsed in three cases when antiseizure medication were decreased or withdrawn after being seizure-free for three to six years, which correlated with the expression stage of FAT1. Genotype-phenotype analysis showed that epilepsy-associated FAT1 variants were missense, whereas non-epilepsy-associated variants were mainly truncated. The relationship between FAT1 and epilepsy was evaluated to be "Strong" by the Clinical Validity Framework of ClinGen. CONCLUSIONS: FAT1 is a potential causative gene of partial epilepsy and febrile seizures. Gene expression stage was suggested to be one of the considerations in determining the duration ofantiseizure medication. Genotype-phenotype correlation helps to explain the mechanisms underlying phenotypic variation.

[Clinical characteristics and prognosis of acute pancreatitis in children]. / å„¿ç«¥æ€¥æ€§èƒ°è ºç‚Žä¸´åºŠç‰¹å¾ä¸Žé¢„åŽåˆ†æž.

OBJECTIVES: To study the clinical characteristics of acute pancreatitis (AP) in children. METHODS: A retrospective analysis was conducted on the children with AP who were hospitalized in the First Affiliated Hospital of Zhengzhou University from January 2020 to June 2022, and their clinical characteristics were summarized and analyzed. RESULTS: A total of 92 children with AP were included, with a male/female ratio of 1:1 and a mean age of (9±4) years. Adolescents (34%, 31/92) and pre-school children (33%, 30/92) were more commonly affected, while infants and toddlers (7%, 6/92) were less commonly affected. The etiology of the disease from most to least was as follows: drug-induced (40%, 37/92), biliary (18%, 17/92), dietary (14%, 13/92), idiopathic (13%, 12/92), trauma-related (9%, 8/92), and infectious (5%, 5/92). Mild, moderate, and severe AP accounted for 68% (63/92), 21% (19/92), and 11% (10/92), respectively. Among all 92 children, 62 (67%) received abdominal ultrasound, with a positive rate of 66% (41/62); 67 (73%) underwent abdominal CT, with a positive rate of 90% (60/67); 20 (22%) underwent magnetic resonance cholangiopancreatography (MRCP), with a positive rate of 95% (19/20). There were significant differences in the levels of D-dimer, procalcitonin, and amylase among children with different degrees of severity of the condition (P<0.05), and there were significant differences in the levels of leukocyte count, hematocrit, blood urea nitrogen, albumin, and blood calcium among children with different etiologies (P<0.05). Of all 92 children, 89 (97%) had a good prognosis. CONCLUSIONS: The primary cause of pediatric AP is medication-induced, with a predominantce of mild cases. Abdominal CT has a high rate of utilization and positivity in the diagnosis of pediatric AP, while MRCP has the highest specificity among imaging techniques. Laboratory tests aid in determining the severity and etiology of AP. The prognosis of AP is favorable in children.

Two cases of Kawasaki disease: Clinical characteristics and onset of fever and gastrointestinal symptoms.

BACKGROUND: Kawasaki disease (KD) is a prevalent form of systemic vasculitis that can damage various organs and systems in children. Typical KD is not difficult to diagnose in clinical practice. In recent years, it has been shown that an increasing number of children do not satisfy the diagnostic criteria for typical KD. This condition is known as incomplete KD (IKD). It is challenging to promptly diagnose and treat such children in clinical practice. CASE DESCRIPTION: A 10-year-old girl was admitted to the hospital due to fever and abdominal pain. She presented with shock symptoms. An enhanced abdominal computed tomography scan revealed intestinal pneumatosis, effusion, and gallbladder enlargement, indicating intestinal obstruction. Due to the poor outcome following an emergency laparoscopic cholecystectomy, IKD was suspected. A 3-month-old male pediatric patient was admitted to the hospital due to a fever. Patchy, congestive rashes formed on the patient's body as KD progressed. IKD was suspected based on the clinical signs of fever, rash, and hyperemia of the lips. The two patients were then treated with human immunoglobulin and aspirin treatment. The prognosis for the two children was favorable following prompt treatment. CONCLUSION: Due to the fact that IKD is frequently misdiagnosed, it is vital to (1) improve the patient prognosis for the early identification of children with KD with prolonged fever and anti-infection failure as the initial manifestation and (2) perform timely diagnosis and comprehensive treatment.

The first case report of McLeod syndrome in an infant with a novel mutation (c.89C>A, p. Ser30X) in XK.

McLeod syndrome (MLS) is a rare multisystem disorder and X-linked recessive inheritance disorder caused by mutations of the X-linked Kx blood group (XK) gene. The manifestations progress slowly and mainly appear in middle age, which make it difficult to distinguish MLS from other neuromuscular disorders. Here, we present a case of a 10-month-old Chinese boy who was taken to hospital for herpes of the extremities and oral cavity along with febrile seizures in June 2017. The laboratory test revealed persistent elevated levels of serum creatine phosphokinase and abnormal liver function. The results of the electrocardiogram showed sinus tachycardia, and magnetic resonance imaging of the brain showed enlarged bilateral ventricles and third ventricle. Genetic analysis by next-generation sequencing revealed a novel nonsense mutation c.89C > A (p. Ser30X) in exon 1 of XK. To the best of our knowledge, this is the first case report of infants with MLS confirmed by genetic analysis.

[Application of next-generation sequencing in the molecular diagnosis of Duchenne muscular dystrophy].

The purpose of this study is to analyze the family's clinical data of 22 children who were given an intended clinical diagnosis of Duchenne muscular dystrophy (DMD), and to explore the clinical value of next-generation sequencing (NGS) in the molecular diagnosis of DMD. The probands were simultaneously tested by NGS for a gene panel associated with hereditary neuromuscular disease and multiplex ligation-dependent probe amplification (MLPA) for the Dystrophin gene. The exon deletion/repetition mutations of the Dystrophin gene determined by both methods were compared and the point mutations of the Dystrophin gene were verified by Sanger sequencing. Dystrophin gene mutations were found in all the 22 probands, including 14 exon deletion/repetition mutations and 8 point mutations/minor variations. The results of MLPA detection were consistent with those of NGS. The results of Sanger sequencing showed that the point mutations and minor variations determined by NGS were correct. One missense mutation (c.6290G>T), 1 nonsense mutation (c.3487C>T) and 4 minor deletion-induced frameshift mutations (c.1208delG, c.7497_7506delGGTGGGTGAC, c.9421_9422delAA and c.8910_8913delTCTC) had not been reported in the Human Gene Mutation Database, and thus were considered as novel mutations of the Dystrophin gene. The results of this study showed that NGS can detect variations in the Dystrophin gene, including exon deletion/repetition, point mutation, minor deletion and intron mutation. Therefore, NGS is of certain clinical value in the molecular diagnosis of DMD and is worthy of recommendation.

Two new compounds from the roots of Scrophularia ningpoensis and their anti-inflammatory activities.

Aiming to investigate the bioactive constituents with anti-inflammatory activity from the roots of Scrophularia ningpoensis, two new compounds (1 and 3) were isolated from the extract of the roots of the plant. Their structures were elucidated on the basis of spectral analyses (UV, IR, NMR, and MS spectroscopy), as well as experimental and calculated electronic circular dichroism (ECD) analyses. All of the isolates were tested for their anti-inflammatory properties in terms of suppressing the production of NO in lipopolysaccharide-induced BV2 cells. Compound 2 exhibited stronger anti-inflammatory effects (77.65%) than the positive control curcumin (69.75%) at 10 µM.

[Change in serum intestinal fatty acid binding protein and its significance in children with pneumonia and gastrointestinal injury].

OBJECTIVE: To study the change in serum intestinal fatty acid binding protein (IFABP) in children with pneumonia and its correlation with gastrointestinal injury. METHODS: A total of 82 children with community-acquired pneumonia who were treated from January to October, 2015 were enrolled, among whom 34 had mild pneumonia and 48 had severe pneumonia. According to pediatric critical illness score (PCIS), the children with severe pneumonia were further divided into non-critical group (25 patients) and critical group (23 patients). Thirty healthy children who underwent physical examination at outpatient service were enrolled as the control group. ELISA was used to measure serum IFABP level, and the acute gastrointestinal injury (AGI) grade was determined for children with severe pneumonia. Serum IFABP level was compared between groups, and the correlations of IFABP with AGI grade and PCIS were analyzed. RESULTS: The severe pneumonia group showed a significantly higher serum IFABP level than the control group and the mild pneumonia group (P<0.01), and the mild pneumonia group also showed a significantly higher serum IFABP level than the control group (P<0.01). The critical group showed a significantly higher serum IFABP level than the non-critical group (P<0.01). The patients with grade I-IV AGI had significantly higher serum IFABP levels than the control group (P<0.01), and the serum IFABP level increased significantly with the increasing AGI grade (P<0.01). Serum IFABP level was positively correlated with AGI grade (P<0.01) but negatively correlated with PCIS (P<0.01). CONCLUSIONS: Children with pneumonia experience an increased serum IFABP level which can be used as a sensitive indicator for the early diagnosis of gastrointestinal injury and the evaluation of conditions in children with pneumonia.

Selective targeting of MAPK family kinases JNK over p38 by rationally designed peptides as potential therapeutics for neurological disorders and epilepsy.

Human mitogen-activated protein kinase (MAPK) family members JNK and p38 are two homologous protein-serine/threonine kinases but play distinct roles in the pathological process of neurological disorders. Selective targeting of JNK over p38 has been established as a potential therapeutic approach to epilepsy and other nervous system diseases. Herein, we describe an integrated in vitro-in silico protocol to rationally design kinase-peptide interaction specificity based on crystal structure data. In the procedure, a simulated annealing (SA) iteration optimization strategy is described to improve peptide selectivity between the two kinases. The optimization accepts moderate compromise in peptide affinity to JNK in order to maximize the affinity difference between peptide interactions with JNK and p38. The structural basis, energetic properties and dynamic behavior of SA-improved peptides bound with the peptide-docking sites of JNK and p38 kinase domains are investigated in detail using atomistic molecular dynamics (MD) simulations and post binding free energy analysis. The theoretical findings and computational designs are then confirmed by fluorescence polarization assays. Using the integrated protocol we successfully obtain three decapeptide ligands, namely RLHPSMTDFL, RAKLPTSVDY and KPSRPWNLEI, that exhibit both potent affinity to JNK (K = 8.0, 5.4 and 12.1 µM, respectively) and high selectivity for JNK over p38 (K/K = 9.2, 17.9 and 6.3 fold, respectively). We also demonstrate that a JNK-over-p38 selective peptide should have a positively charged N-terminus, a polar central region and a negatively charged C-terminus, in which a number of hydrophobic residues distribute randomly along the peptide sequence. In particular, the residue positions 1, 6 and 9 play a crucial role in shaping peptide selectivity; the presence of, respectively, Arg, Thr and Asp at the three positions confers high specificity to kinase-peptide interactions.

Elevated interictal serum galectin-3 levels in intractable epilepsy.

BACKGROUND: Intractable epilepsy is defined as the occurrence of seizures that cannot be controlled with medical treatment. The discovery of epilepsy biomarkers is increasingly attracting more attention from both clinical physicians as well as neuroscientists. Increased levels of soluble and/or cellular galectin-3 (Gal-3) have been associated with various diseases. However, the effects of Gal-3 in epilepsy are still unknown. In this study, we evaluated the association of higher interictal serum Gal-3 protein levels in patients diagnosed with intractable epilepsy. PATIENTS AND METHODS: A group of 38 patients with intractable epilepsy and 26 healthy age-matched control subjects were included in this study. A commercially available electrochemiluminescence immunoassay (ECLIA) kit was used to determine serum Gal-3 protein levels. RESULTS: Our results indicated that serum Gal-3 protein level in the patient group was 6.67 ± 0.34 ng/ml, and in the age-matched control group was 5.40 ± 0.34 ng/ml. The difference between the two groups was found to be statistically significant (P = 0.003). CONCLUSION: This study found a detectable elevation in serum Gal-3 concentration in patients with focal epilepsy. Given its secretory nature and detectable levels in the serum, Gal-3 could be a potential biomarker for intractable epilepsy.

[Hydroa vacciniforme-like cutaneous T cell lymphoma: a case report and literature review].

OBJECTIVE: To study the clinical features, diagnosis and therapy of hydroa vacciniforme-like cutaneous T cell lymphoma. METHODS: The clinical presentations and the findings of laboratory examinations and skin biopsy of affected tissue in a child with hydroa vacciniforme-like cutaneous T cell lymphoma were retrospectively reviewed. RESULTS: The child manifested as rash, fever and lymph node intumesce. Rash was pantomorphia, including edematous erythema, vesicles, crusts, necrosis and depressed scar, and it was mild in winter and severe in summer, mainly involving in the face and extremities. Epstein-Barre virus (EBV)-IgM was positive. Histopathological findings revealed focal lymphocyte invasion in subcutaneous panniculus adiposus, mainly surrounding the blood vessels. Immunohistochemistry showed CD3 (+), CD43 (+), CD20 (-), pax-5 (-), TIA (+), CD5 (+), CD8 (+), Granmye (+) and CD4 (-). The clinical symptoms were improved after glucocorticoid treatment in this child. CONCLUSIONS: Hydroa vacciniforme-like cutaneous T cell lymphoma has special clinical manifestations. This disorder may be definitely diagnosed by skin biopsy of affected tissue and immunohistochemistry assay. Glucocorticoid treatment is effective. EBV infection may be related to the development of this disorder.

[Ten-year changes in pathogen, antimicrobial susceptibility and clinical feature of children with bacterial meningitis].

OBJECTIVE: Despite progress in antibiotic therapy and intensive care, childhood bacterial meningitis (BM) remains a devastating disease. We conducted this study to investigate the changes in clinical characteristics, the etiologic agents and antimicrobial susceptibility of BM during the past 10 years in children under 14 years of age. METHODS: These 126 patients were divided into two groups according to their date of admission. Group 1 included 64 patients admitted from January 1998 to December 2002, and group 2 included 62 cases admitted from January 2003 to December 2007. All pediatric medical charts of them were reviewed. RESULTS: The predominant isolated bacteria from CSF were coagulase-negative staphylococcus (17/62, 27.4%) and Escherichia coli (9/62, 14.5%) in group 2. The resistance rate of staphylococcus against oxacillin (MRS) was 68.4% (13/19) in group 2, significantly higher than that of group 1 (16.7%, 2/12). Among 126 cases, 42 had seizure attack and 16 had consciousness disturbance, the proportions of them in group 2 (11/62, 17.7%; 4/62, 6.4%) were lower than those in group 1 (31/64, 48.4%; 12/64, 18.8%, P < 0.05). Cases in group 2 survived with complications [13/62 (21.0%)] and sequelae [11/62 (17.7%)] were lower than those in group 1 (24/64, 37.5%, 23/64, 35.9%, P < 0.05), but the rate of empirical therapy modification in group 2 (21/62, 33.9%) was higher than that in group 1 (7/64, 10.9%, P < 0.01). CONCLUSION: The predominant bacteria in children with BM are staphylococcus and Escherichia coli in recent years. The antibiotic resistance rate of bacteria has been higher year after year. The clinical patterns of pediatric BM have changed with a decrease in clinically serious cases, complications and sequelae, but an increase in modification of empirical therapy.

[Effect of a novel splicing mutation (IVS2-2A-->C) of SEDL gene on RNA processing].

X-linked spondyloepiphyseal dysplasia tarda (SEDL) is a rare osteochondrodysplasia caused by the mutation of SEDL gene, which mainly involves vertebral bodies and hips. To explore the effect of the novel splicing mutation (IVS2 -2A-->C) of SEDL gene on mRNA processing in a large Chinese family with X-linked spondyloepiphyseal dysplasia tarda and to elucidate the molecular base of SEDL, total RNA was isolated from EDTA blood samples of patients and controls. RT-PCR was performed on total RNA. cDNA was analyzed by bi-directionally direct sequencing of PCR products and Polyacrylamide gel electrophoresis (PAGE). Sequencing analysis revealed that there were two kinds of cDNA in patients. One is that exon 2 directly spliced exon 4, that is, exon 3 absence from the mature mRNA; and the other is that exon 1 directly spliced exon 4, meaning both exon 2 and 3 being spliced out completely. Meanwhile one kind of cDNA that exon 1 directly spliced exon 3 was found in normal controls. By PAGE, RT-PCR amplified products, 679bp and 537bp, were detected in normal controls, while 567bp and 425bp fragments were found in affected individuals. Our data show that the mutation of the splice-acceptor site in intron 2 causes exon 3 entirely exclusion from the mature RNA transcripts in affected individuals. As the translation start site of the SEDL gene locates on exon 3, the splicing defect causes affected individuals failure to produce sedlin, which elucidates the causative role of SEDL gene in the pathogenesis of SEDL. The absence of exon 2 indicates that there is alternative splicing in SEDL gene. The alternative splicing was also found in normal controls, which demonstrated that the alternative splicing might not be related to the phenotype of SEDL. Because the alternative splicing of exon 2 occurred in the 5'UTR, it is not clear whether it affects the gene expression.

[Clinical study of 83 cases with spinal muscular atrophy in children].

OBJECTIVE: Spinal muscular atrophy (SMA) is a common autosomal recessive disorder and represents one of the most common genetic causes of death in childhood. The last 10 years have seen major advances in the field of SMA, but no curative treatment is available so far. This study aimed to analyze the clinical characteristics of SMA, improve the clinical diagnosis of SMA, and explore the importance of gene diagnosis and prenatal diagnosis of SMA by gene deletion analysis. METHODS: Totally 83 cases with SMA including 55 males and 28 females were enrolled in this study. The age was between 1 day and 14 years (average 23.7 months). The clinical characteristics and changes of electromyography were assessed in all cases. The muscular biopsy was performed in 2 of 83 cases. The deletion of survival of motor neuron gene (SMN) was detected by PCR and restriction endonuclease spectrum analysis in 13 of 83 cases. RESULTS: The 83 cases were subdivided into three clinical groups based on age of onset of symptom, age at death and achievement of certain motor milestone, 60 cases with type I, 19 cases with type II and 4 cases with type III. They were all characterized by symmetric muscle weakness (more proximal than distal) associated with atrophy, absence or marked decrease of deep tendon reflexes. Electromyographic studies showed a pattern of denervation with neither sensory involvement nor marked decrease of motor nerve conduction velocities in all cases. Muscle biopsy provided evidence of skeletal muscle denervation with groups of atrophy in 2 cases. The SMN detection revealed deletion of exon 7 and exon 8 in 11 of 13 cases, only lacking exon 7 in 1 of 13 cases and lacking exon 8 in 1 of 13 cases. CONCLUSION: SMA is characterized by degeneration of lower motor neuron associated with muscle paralysis and atrophy. The definite diagnosis of SMA will rely on the typical clinical characteristics, changes of electromyogram and muscle biopsy and gene deletion analysis. Gene diagnosis of SMA can provide a basis for prenatal diagnosis which is of great importance in preventing SMA.

[Identification of a novel mutation IVS2-2A-->C of SEDL gene in a Chinese family with X-linked spondyloepiphyseal dysplasia tarda].

OBJECTIVE: To identify the mutation of spondyloepiphyseal dysplasia tarda (SEDL) gene in a large Chinese family with X-linked spondyloepiphyseal dysplasia tarda and to make a discussion on the pathogenesis of SEDL at the molecular level. METHODS: In two patients, four exons comprising the SEDL open reading frame as well as their exon/intron boundaries were analyzed by bi-directional direct sequencing of PCR products. The sequencing results were compared against the normal sequences in GenBank to find the mutation. Then the mutation was identified in other members of the family. RESULTS: A nucleotide substitution of the splice acceptor in SEDL intron 2, IVS2 -2A-->C,was detected in two affected individuals (IV(15) V(3)) in the Chinese family with SEDL, but no sequence change occurring on exons 3-6 was detected. The transversion was also identified in four heterozygous carriers. The mutation was not found in two unaffected male individuals and fifteen normal controls. Furthermore, four potential carriers were identified in the family. CONCLUSION: The mutation IVS2 -2A-->C of SEDL gene was firstly determined in the world. The change of the splice acceptor in SEDL intron 2 may cause skipping of exon 3 which is responsible for the disease. Molecular diagnosis can be made by detecting the mutation.

[Gene diagnosis of X-linked spondyloepiphyseal dysplasia tarda by linkage analysis and DNA sequencing].

OBJECTIVE: X linked spondyloepiphyseal dysplasia tarda (SEDL) is heritable osteochondrondysplasia characterized in affected males by disproportional short stature with short neck and trunk resulting from a growth defect of the vertebral bodies, accompanied by barrel chest and degenerative osteoarthropathy of hip joints. This progressive skeletal dysplasia is caused by the SEDL gene located approximately 100 kb centromeric of DXS16 at Xp22. The disorder usually manifests in late childhood without systemic complications, and generally female carriers of SEDL are asymptomatic. So the diagnosis of potential carriers and presymptomatic patients is almost impossible. This study aimed to establish methods of gene diagnosis for finding out potential carriers and presymptomatic patients. METHODS: The blood samples were collected from 21 individuals in a large Chinese pedigree with SEDL. Microsatellite marker DXS16 was selected for linkage analysis. In order to confirm the allele of DXS16 linked to the pathogenic SEDL gene, polymerase chain reaction (PCR) and polyacrylamide gel electrophoresis (PAGE) were used to examine the variability of the lengths of DXS16, and linkage analysis was performed for the diagnosis of potential carriers and presymptomatic patients. Then the pathogenic mutation of the SEDL gene in the family was identified by bi-directionally direct sequencing of PCR products amplified for each of the four coding exons as well as their exon/intron boundaries. The potential carriers and presymptomatic patients were also diagnosed in this way. RESULTS: Six young individuals (IV(14), IV(19), IV(21), IV(23), V(4), V(7))who wanted to know whether they were carriers or presymptomatic patients were diagnosed by linkage analysis. Four females of them (IV(14), IV(19), IV(21), V(7)) were determined being carriers because they carry the allele of DXS16 which links the pathogenic SEDL gene, and the other two (IV(23), V(4)) being normal individuals for their alleles of DXS16 linked with wild SEDL gene. DNA sequencing identified that the pathogenic mutation of SEDL gene in the family, which was a nucleotide substitution of the splice-acceptor site in intron 2, IVS2 -2 A-->C. This is a novel mutation in the SEDL gene. Four female individuals (IV(14), IV(19), IV(21), V(7)) carried the mutation; individuals IV(23) and V(4) carried the wild SEDL gene. The results of diagnosis of linkage analysis coincide completely with that of DNA sequencing. CONCLUSION: Linkage analysis is a simple, rapid and inexpensive gene diagnosis method for SEDL and its accuracy was the same as DNA sequencing. Each of linkage analysis and DNA sequencing can be used to diagnose SEDL, which is very helpful for finding potential carriers and presymptomatic patients.