Objective To compare the diagnostic accuracy of mass spectrometry and high throughput sequencing in screening neonates at high risk of suspected inherited metabolic disorders (IMD). Methods Neonates with suspected IMD were referred to mass spectrometry and next generation sequencing (NGS) with the consent of their guardians from August 2016 to June 2018. Taking NGS (containing 2 742 known pathogenic genes) as the gold standard, the diagnostic value of mass spectrometry (screening diseases including 20 amino acid metabolic diseases, 12 fatty acid metabolic diseases and 19 organic acid diseases) for IMD was discussed. Results Thirty-three of 2 000 neonates with suspected IMD were diagnosed by NGS, including 8 kinds of abnormal organic acid metabolism(15 cases), 7 kinds of abnormal amino acid metabolism(9 cases), 3 kinds of abnormal fatty acid oxidation metabolism(6 cases) and 3 cases of other types of IMD. In these patients, mass spectrometry indicated that there were 128 cases of secondary changes and 26 cases of IMD. When both IMD and secondary changes were considered positive according to the results of mass spectrometry, the sensitivity and specificity of mass spectrometry were 93.9% (95%CI: 79.8%-99.3%) and 93.8% (95%CI:92.6%-94.8%) respectively. In special types of IMD indicated by mass spectrometry, there were 13 cases consistent with NGS, but 2 cases were inconsistent with NGS. Indicated by mass spectrometry, NGS detected pathogenic/likely pathogenic variants of related IMD in 11 cases with several alternatives of IMD, 5 cases with secondary changes and 2 cases with normal results. Five IMDs identified by NGS were not included in the disease spectrum of mass spectrometry in our study. Among the patients with IMD, 12 had family history of suspected IMD. In the 6th month after birth, 13 cases with IMD died of abandoned treatments and 2 cases died of invalid rescue. NGS established therapeutic schemes for 18 infants with IMD. In these patients, 7 developed well and 11 had developmental delay, mainly psychomotor retardation, some of which had hearing impairment. Conclusion Compared with mass spectrometry, the combination of mass spectrometry and NGS can diagnose neonates of suspected IMD more quickly so as to implement individualized treatment as early as possible and improve the prognosis of patients.
Objective To investigate the etiology and prognosis of epileptic encephalopathies(EEs) in children. Methods We reviewed the clinical data of patients with EEs in childhood to analyze their etiologies. The therapeutic effect and development outcome were followed up. Results We enrolled 234 cases of EEs, including West syndrome (n=92), Dravet syndrome (n=53), Lennox-Gastaut syndrome (n=32), Landau-Kleffner syndrome (n=15), Ohtahara syndrome (n=13), epileptic encephalopathy with continuous spike-and-wave during sleep (n=10), myoclonic-atonic epilepsy (n=2), Rasmussen syndrome (n=2), and the unclassified 15 cases. Genetic etiology accounted for 67.9% (31.2% confirmed by gene detection), structural for 12.0%, infectious for 4.7%, metabolic and immunogenic for 0.9% respectively, and 19.2% remained unknown. The effective rates of antiepileptic drugs (AEDs), ketogenic diet, surgery and vagus nerve stimulation (VNS) were 33.8% (79/234), 47.3% (35/74), 60.0% (3/5) and 44.4% (4/9) respectively.The effective rate of comprehensive treatment (drug treatment alone or ketogenic diet/surgery/VNS based on drug treatment) was 48.7%. Development outcome was that 223 cases (95.3%) showed developmental retardation in the first intelligence assessment after onset and 215 cases (91.9%) showed developmental retardation after treatment (median follow-up duration: 44 months). Conclusion Genetic etiology was the primary cause of EEs. AEDs were effective in only one-third of cases, and the incidence of developmental retardation was high, leading to poor prognosis. Multidisciplinary diagnosis and treatment (MDT) played a key role in the diagnosis and management of EEs.
Objective To investigate the clinical features, diagnosis and treatment of inherited coagulation factor Ⅶ deficiency (FⅦD) in newborns. Methods The clinical manifestations, diagnosis and treatment process, and genetic testing of two cases of neonatal FⅦD diagnosed at Guangzhou Women and Children's Medical Center were retrospectively analyzed. The literature about the clinical features of FⅦD in the neonatal period was reviewed and summarized. Results Two female cases of FⅦD presented with severe gastrointestinal tract and intracranial hemorrhage. Their laboratory tests were characterized by repeated and non-VitK1-dependent prothrombin time (PT) prolongation and normal activated partial thromboplastin time (APTT). The factor Ⅶ activity was 1.5% and 3% respectively. These two cases both died of a large area of intracranial hemorrhage at 31 days and 6 months after birth respectively. Gene sequencing results showed a homozygous mutation in the F gene IVS7+1G>T cleavage site in both two cases. In addition, we reviewed 24 cases of full-term neonatal FⅦD of both sexes. Among them, 22 cases were reported by the literature, and 2 by this report. Most of newborns(73.7%) had the first onset of the disease mostly within 7 days after birth. The main initial symptoms of disease onset included gastrointestinal bleeding (blood stool, vomiting blood) (9/24), nervous system signs (drowsiness, convulsions, poor response) (8/24) and pale skin(7/24) .The proportion of intracranial hemorrhage was high(23/24). PT was significantly prolonged, and FⅦ activity was significantly decreased(≤ 5% accounting for 83.3%). The incidence of mortality and disability was 70.8%(17/24). Fourteen of 24 babies got positive F gene diagnosis. Conclusion FⅦD is a rare disease in the neonatal period. Once the disease occurs, life-threatening bleeding is prone to occur and the prognosis is always poor. FⅦD can be clinically confirmed with the presence of prolonged PT that is difficult to correct and significantly decreased FⅦ activity. As mutations in some sites of F gene can cause severe bleeding, a genetic diagnosis contributes to prenatal diagnosis.
Objective To improve the prenatal diagnosis accuracy of anomalous origin of one pulmonary artery branch from the aorta (AOPA) by analyzing fetal echocardiography features. Methods Fetal echocardiographic features were analyzed retrospectively in 6 cases with prenatal diagnosis of AOPA from June 2012 to October 2018, and the fetal echocardiography characteristics of AOPA were summarized. Results Among the 6 fetuses with AOPA, 5 cases were anomalous origin of right pulmonary artery from the aorta (AORPA), including 3 cases with isolated AORPA and 2 cases combined with Berry syndrome, and one case was anomalous origin of left pulmonary artery from the aorta (AOLPA) with tetralogy of Fallot (TOF) with an absent pulmonary valve. All cases had proximal form. Among the 6 fetuses, 2 cases had induced labor, including one case with TOF and one case combined with Berry syndrome. One case with Berry syndrome died of recurrent pneumonia and heart failure at 63 days after birth without surgical treatment. Other 2 cases with isolated AORPA underwent surgical repair after birth and recovered well after surgery. One case was just born and still under observation. Characteristic sonographical findings included: ① The main pulmonary artery was continuous with one pulmonary artery and there was no confluence with the other pulmonary artery. The diameter of normal pulmonary artery branch was widened. ② On the basis of three vessels and trachea view (3VT) and long axial view of left ventricle (LV), two-dimensional and color Doppler imaging (CDFI) showed a large vessel originated from the aortic arch, and the anomalous vessel arose proximally from the right or left lateral posterior aspect of the ascending aorta close to the aortic valve. ③ CDFI showed that the blood flow of the anomalous pulmonary artery branch was isolated and there was no exact connection with the main pulmonary artery. ④ Spectrum Doppler of the anomalous vessel of the lung confirmed that it was the anomalous origin of one pulmonary artery branch. ⑤ When other intracardiac abnormalities were not present, the inner diameter and proportion of the heart cavity were normal. Conclusion Proximal form of AOPA in the prenatal period can be accurately diagnosed by echocardiography. The 3VT and long axial view of LV are very important for the diagnosis of proximal form of AOPA.
Objective We aimed to report the clinical manifestations, immunological features, genetic diagnosis and treatments of two cases with LRBA (LPS-responsive beige-like anchor protein) gene mutation. Methods Two male cases with LRBA gene mutations were enrolled in our study. The clinical features, immunological features and treatments of the two patients were retrospectively summarized. The lymphocyte subsets were detected by Flow Cytometry. WES was adapted for genetic analysis and the LRBA gene mutations were confirmed by Sanger sequencing，while the deletion of a gene was confirmed by fluorescence quantitative PCR. Results Patient 1 (P1) presented with recurrent throbocytopenia and hematocytopenia for six years. He also had recurrent fungal infection, persistent EBV viraemia, growth retardation and apparent hepatosplenomegaly. Patient 2 (P2) presented with recurrent thrombocytopenia after birth. They were diagnosed at the age of 13 years and 4 months respectively. Both of them presented with increased naive B cells and decreased memory B cells. P1 presented with decreased levels of immunoglobulin(Ig) M, IgG and IgA. Decreased naive CD4+ T and CD8+ T cells, expanded central memory CD4+ T and CD8+ T cells, reduced CD4+ T lymphocytes, B lymphocytes and NK cells, and increased CD8+ T cells were also observed in P1. P2 presented with increased IgG level. Sequencing results revealed that P1 had c.1933C>T (p.R645X) mutation and exon 29 deletion in LRBA gene. P2 had compound heterozygous mutations in LRBA gene (c.3778G>C, p.A1260P and c.1570G>A, p.G524S). Finally, P2 underwent hematopoietic stem cell transplantation (HSCT) and was currently in remission. Meanwhile P1 was under symptomatic treatment. Conclusion LRBA deficiency has a wide spectrum of phenotypes, characterized by hypogammaglobulinemia, recurrent infections, autoimmune diseases and susceptibility to EBV infection. Intravenous immune globulin could help relieve infection. Abatacept targeted therapy and HSCT might be the availabe therapeutic options for patients with LRBA deficiency.
Objective We aimed to analyze the onset age and genotype of CHD2-related epileptic encephalopathy and to improve the understanding that the disease can occur in the neonatal period. Methods We analyzed one newborn with epileptic encephalopathy admitted to Children's Hospital of Fudan University. We searched the gene "CHD2" in HGMD database and key words including "CHD2 gene" in CNKI, VIP database, Wanfang database, PubMed and the Web of Science database. All of the databases were searched up to Nov 29, 2018. Results The male newborn in the hospital mainly presented with seizures, dystonia and feeding difficulties. A de novo mutation［NM_001271:exon31:c.3951G>C(p.L1317F)］was detected in CHD2 gene. We reviewed 67 cases from Chinese and foreign literature databases. None of the reported cases had seizures during the neonatal period. Sixty-one point one percent (11/18) of cases showed developmental delay before the onset of seizures. According to the reported cases and the case from this study, we found that all of the three cases who presented with seizures within 6 months (including 6 months old) carried missense variants of CHD2 (3/3，100%), and 5 of 19 cases (5/19，26.3%) occuring seizures at the age of older than 6 months carried missense variants. Conclusion The defected CHD2 can cause epileptic encephalopathy in the neonatal period. The missense variants of CHD2 may have a tendency to cause early epilepsy. Defected CHD2 should be taken into consideration for patients with neurodevelopmental disorders and no seizures.
Objective To analyze the effects of preterm birth and low birth weight on infant development, and to explore the potential interaction effect of preterm birth and low birth weight on the development of infants. Methods Data of infants at the age of 8 to 10 months receiving Denver Development Screening Test (DDST) from September 2013 to December 2017 in Tongzhou District of Beijing City were collected through the electronic information system of Maternal and Child Health Care Hospital of Tongzhou District. Using the method of nested case-control study, the children with DDST screening results of "suspicious" or "abnormal" were taken as the case group, and the children with DDST screening results of "normal" were taken as the control group. Chi-square test was used to analyze the development of infants and its influencing factors. Univariate and multivariate unconditional logistic regression models were used to analyze the possible influence of preterm birth and low birth weight on infant development. The interaction effect of preterm birth and low birth weight on infant development was investigated by using the additive and multiplicative interaction models. Results A total of 22 455 infants aged from 8 to 10 months were included in this study and the prevalence of suspicious/abnormal DDST results was 2.6%. The infant with younger gestational age or lower birth weight might have higher risk of suspicious/abnormal DDST results at the age of 8 to 10 months. In interaction analysis, preterm birth and low birth weight were observed to have additive interaction to the infant development (RERI=2.55, 95%CI: 0.54-4.55), but their multiplicative interaction was not observed (OR=1.50，95%CI：0.68-3.30). Conclusion There was an additive interaction between preterm birth and low birth weight, and no multiplicative interaction was found between these two factors. It should be paid more attention on the early development of infants with preterm birth and low birth weight.
Objective The clinical features of gynecomastia in children and adolescents under 18 years old were analyzed retrospectively to explore the main influencing factors of gynecomastia in children and adolescents. Methods This study was a case-control study. The male patients with gynecomastia as the chief complaint and without drug intervention at the department pediatric endocrinology of the First Affiliated Hospital of Zhejiang University were enrolled into the gynecomastia group. The control group consisted of healthy boys of the same age. Height and weight were measured; body mass index (BMI) was calculated; sex hormone levels were detected by chemiluminescence immunoassay. Results The mean age of gynecomastia group (n=298) was (12.4±2.8). Gynecomastia reached its peak around the age of 11, and then showed a decreasing trend. G2 and G3 stage of sexual development accounted for 65.4%. The mean age of the control group (n=197) was (12.1±2.5). Logisitic regression analysis showed that BMI, T, FSH, LH, PRL and E2 / T were the risk factors of the gynecomastia. The sensitivity of the six indicators in combination was 0.748, and the area under the curve (AUC) was 0.813 (95% CI: 0.767-0.858). Conclusion The combination of BMI, T, E2/T, FSH, LH and PRL had a good predictive value for gynecomastia.
ObjectiveTo investigate the clinical and molecular characteristics of hospital-associated (HA) infections caused by methicillin-resistant Staphylococcus aureus (MRSA) in pediatric patients. MethodsWe collected clinical S. aurues isolates from the patients (＜14 years old) with HA-MRSA infections from 2010 to 2016 at Beijing Children's Hospital. The medical records were retrospectively reviewed and collected. Isolates as HA-MRSA were further characterized by MLST, SCCmec and spa typing methods. Carriage of pvl genes were also detected. Antibiotic susceptibility test was performed to detect drug resistance to antibiotics. ResultsA total of 150 cases were collected, with a median age of 18 months. Pneumonia (55.3%) was the most common disease, followed by skin and soft tissue infections (46.0%). A total of 16 ST genotypes were obtained with common clones of ST239 (46%) and ST59(28%). Three SCCmec types were detected and SCCmecⅢ was the predominant one, accounting for 52.7%. Thirty-two spa types were detected and t030 (22.0%) and t437 (23.4%) were the top two clones. The multidrug resistance rate for all isolates, MRSA-ST59 and MRSA-ST239 were 84.0%, 78.6% and 81.2% respectively. ConclusionHospital-associated MRSA infections were common in young children, and the genetic background was diverse. The most common clones were ST239-SCCmecⅢ-t030 and ST59-SCCmecⅣ-t437. The rate of multiple drug resistance was high in the isolates.
ObjectiveTo explore the potential role of LINC01503 in the regulation of macrophage polarization and its function in the pathology of children asthma to achieve a better understanding of the pathology of asthma and find new strategies for asthma treatment. MethodsBlood was collected from asthmatic patients and healthy controls from August 26, 2017 to January 11, 2018 at Children's Hospital of Fudan University. PBMCs were separated from blood by gradient centrifuge. RT-qPCR was used for the detection of the expression of LINC01503 in PBMCs. LPS and IL-4 were used for the inducing of M1 and M2 polarization of macrophage separately. siRNA was used for knocking down LINC01503. RT-qPCR and Western blot were used for the detection of the effect of LINC01503 on the regulation of macrophage polarization. ResultsWe enrolled 28 patients of asthma and 46 healthy controls. Comparing with healthy controls，the expression of LINC01503 was significantly enhanced in asthmatic patients. LINC01503 was up-regulated in M2 macrophage but down-regulated in M1 macrophage. LINC01503 inhibited the M2 macrophage polarization indicated as the down regulation of CD206 and CD209. On the opposite, LINC01503 promoted the M1 macrophage polarization indicated as the up regulation of the expression of inflammatory mediators such as IL-6, TNF-α and CXCL10 through enhancing the phosphorylation of ERK. ConclusionLINC01503 was up-regulated in the asthmatic patients and it regulated macrophage polarization through a negative feedback loop.
ObjectiveTo observe the expression of mucin-1 (MUC1) and mucin-2 (MUC2) in intestinal tissue of neonatal necrotizing enterocolitis (NEC). MethodsThis is a case-control study. Ten intestinal tissue samples of neonates with NEC at Children's Hospital of Chongqing Medical University were selected randomly as NEC group and their general clinical data were collected. Neonates with congenital intestinal atresia were matched 1∶1 with NEC group mainly based on gestational age and day of disease onset. All the samples were tested by HE-staining and immunohistochemistry methods. ResultsThe structure of intestinal tissue with NEC was significantly different from that in the control group. The former was mainly demonstrated as rupture of the integrity of intestinal tissue. Expression levels of MUC1 and MUC2 in intestinal tissue of neonatal NEC were both significantly decreased compared with the controls (statistically analyzed by integrated optical density, IOD). The median of MUC1 IOD was 780 455.5 in NEC group, and 19 175 070.4 in the control group, P=0.004. The median of MUC2 IOD was 3 039 120 in NEC group, and 45 750 707.5 in the control group, P=0.001. ConclusionMUC1 and MUC2 were key elements of intestinal mucosal barrier and may take part in the pathogenesis of NEC.
ObjectiveTo provide the distribution of different etiologies in pediatric hemoptysis in China and abroad.MethodsChinese databases (Wanfang Data, CBM, VIP and CNKI) and English databases (PubMed, Cochrane Library and Embase) were searched for the retrospectively observational studies based on the criteria, which had been done by 2 independent reviewers up to January 1st, 2018. The quality of enrolled studies was assessed according to CARE checklist. The meta-analysis was performed by Stata 14.1.ResultsA total of 11 studies met the inclusion criteria, and a total of 523 children were involved. It was demonstrated that pneumonia and bronchitis were the most common causes(36.4%), and the other common causes included idiopathic pulmonary hemosiderosis (IPH, 10.3%), pulmonary tuberculosis (5.0%), cardiopulmonary vascular malformations (5.8%), bronchiectasis (5.7%), bronchial foreign bodies (1.9%), trauma (1.3%), other causes (10.4%) and unclear etiologies (9.1%). Outcomes in meta-analysis demonstrated that in total the prevalence rate of respiratory infection was 39.8% (95%CI: 35.4%-44.1%), and 8.3% (95%CI: 4.6%-12.1%) for IPH, 4.5% (95%CI: 2.5%-6.5%) for pulmonary tuberculosis, 4.9% (95%CI: 2.7%-7.1%) for bronchiectasis, and 9.0%(95%CI: 4.9%-13.2%) for cardiopulmonary vascular malformations. The distribution of etiologies was proved different in China.ConclusionThe causes leading to hemoptysis in children were different from those in adults, and the most common causes were infectious diseases, IPH and cardiopulmonary vascular malformations. Additionally, about 20% of pediatric hemoptysis was caused by other diseases and unclear conditions.
