AbstractObjective: To determine the reliability and validity of Chinese version of Mini Manual Ability Classification System(Mini-MACS) for Cerebral Palsy. Methods: Inclusion criteria: Children diagnosed as cerebral palsy aged from 1 to <4 in Children's Hospital of Fudan University and Shanghai cooperation hospitals of children's rehabilitation cooperation group from October to November 2017. Exclusion criteria: Children with severe hearing and visual impairments. The test retest reliability of Mini-MACS was determined by analyzing the relationship between site operation evaluation and video evaluation results. The Mini-MACS inter rater reliability was determined by analyzing different inter rater (caregivers, occupational therapists and physical therapist). By analyzing the correlation of the fine motor ability scores of Fine Motor Function Measure scale(FMFM), the validity of Mini-MACS was determined.Results: 76 children with CP were included, of them 18 cases were from Children's Hospital of Fudan University, 58 cases were from Shanghai children's rehabilitation group cooperation units; 46 male, 30 female; average age was（2.5±0.9） months, 25 cases aged 1 to 2 years, 23 cases aged~<3 years, 28 cases aged ~4 years; 29 spastic quadriplegia, 11 double paralysis, 27 hemiplegia, 2 athetosis, 2 ataxia type, and 5 could not be classified; 7 of them Gross Motor Function Classification System(GMFCS) grade Ⅰ, 27 grade Ⅱ, 16 grade Ⅲ, 16 grade Ⅳ, 10 grade Ⅴ. The results of OT's site evaluation and video evaluation showed that the Mini-MACS had a good retest reliability, and the ICC value was 0.96 (95% CI: 0.94-0.98). At the same time, the mini-MACS had a good inter rater reliability. The ICC value between OT and PT was 0.97 (95% CI: 0.94-0.98), and the ICC value of OT and primary caregivers was 0.92 (95% CI: 0.86-0.95). Among 1~2, 2~3 and 3~4 years old group, all three age groups showed high retest reliability (ICC value between 0 remarks ~1.00), and 3 ~4 inter rater reliability between the evaluators was lower than 1~2 and 2~3 years old groups; In particular, the reliability between OT and primary caregivers was significantly lower than that of the other two groups (ICC= 0.64). There was a good parallel validity between Mini-MACS and fine motor ability score, and Spearman correlation coefficient was -0.76.Conclusion: The Chinese version of Mini MACS has a good inter rater reliability and parallel validity. It is suitable for the classification of younger children with cerebral palsy.
AbstractObjective: To evaluate the association between the antibiotic use during pregnancy and childhood asthma. Methods: The Chinese and English databases CNKI, Wanfang Data，Chinese VIP science and technology periodical database, PubMed, EMBASE, Cochrane and ovid were searched for cohort studies on the association between the use of antibacterial agents in pregnancy and childhood asthma. This meta-analysis with Stata12.0 software explored the relationship between antibiotic and asthma.Results: There were 9 studies with the prevalence of childhood asthma and asthma exposure results related factors (OR, RR or HR) and were adjusted for maternal asthma, smoking and other confounding factors.The results of the meta-analysis showed that the use of antibacterial agents in pregnancy increased the risk of childhood asthma (OR=1.14, 95%CI: 1.13-1.15).Because of the high heterogeneity(I2=96.5%),analysis of the heterogeneity among literatures was carried out: the sensitivity analysis was made by removing each article,and the pooled risk estimates of asthma (OR=1.27, 95% CI: 1.17-1.38, I2=0）after excluding the studies of high heterogeneity that did not affect the final result;analysis of clinical heterogeneity: the subgroup analysis with the age of 3 years of asthma onset was performed ,and the pooled effect size OR of studies with < 3 years and ≥3 years respectively was 1.85(95%CI: 0.80-4.29, I2=78.8%) and OR=1.19 (95%CI: 1.08-1.31, I2=96.8%); stratified by trimesters of pregnancy, the OR was 1.29(95%CI: 1.23-1.34), 1.30(95%CI: 1.25-1.35) and 1.26(95%CI: 1.21-1.31) for the early-, mid-and late- pregnancy, respectively, and I2 =0; stratified by antibiotic assessment, the pooled OR from self reporting(interviews) and from the database was 1.27 (95%CI: 1.10-1.48, I2=71.2%), and 1.20 (95%CI:1.08-1.32, I2=98.6%) respectively; stratified by antibiotic tepy,the OR was 1.18（95%CI: 1.08-1.30，I2=0） for Beta- lactam antibiotics and 1.19（95%CI: 0.78-1.83，I2=83.5%）for Sulfonamides and trimethoprim. ②In sibling analyses, the pooled OR was 0.91 (95%CI: 0.79-1.06, I2=93.3%). There was no publication bias by the Begg rank correlation test and the Egger linear regression test.Conclusion: This meta-analysis suggested that antibiotic exposure during pregnancy may increase the risk of asthma in offspring.But in the sibling control analysis the associations disappeared, indicating the residual factors such as family environment,heredity and so on were important confounding factors that may lead to reverse causality. Therefore, the causal relationship between antibiotic use during pregnancy and risk of asthma in offspring needs further investigation.
AbstractObjective: To evaluate the effects of very preterm birth (born≤32 weeks of gestation) and/or very low birth-weight (VLBW, birth weight≤1 500 g) on brain volume development during adolescence of 13 to 18 years old. Methods: Databases including PubMed, EMBASE, the Cochrane Library, Web of Science, Chinese Biomedical Database (CBM), WanFang Data and CNKI from inception to October 20, 2017 were searched for studies that reported volumetric outcomes in adolescents using magnetic resonance imaging, including cohort studies in which experimental groups were born with gestational age ≤32 weeks and / or birth weight ≤1 500 g, control group with gestational age of 38 to 42 weeks and birth weight ≥2 500 g, and excluding studies with congenital malformations or diseases at birth, presence of disease that may affect measurement outcome during follow-up, and studies failure to extract the measurements herein. The outcome indicators were whole brain, white matter and gray matter, cerebellum, hippocampal and corpus callosum volume. The quality of the literature was evaluated with Newcastle-Ottawa scale (NOS) and meta-analysis was conducted with R 3.4.0 software, and the heterogeneity among the studies was assessed by I2 test. Results: Thirteen English studies (n=1 272) entered the meta-analysis, including 686 cases in the experimental group and 586 cases in the control group. None of the 13 studies was followed completely. In 9 studies non-exposed and exposed groups were not from the same community. 3 studies were only based on retrospective questionnaire. All of the 13 studies used MRI with field strength of 1.5T, computer software to measure the volume automatically, and the same endpoint age for both cohorts. Meta-analysis showed that compared with the control group, the whole brain volume (SMD=-0.66, 95%CI: -0.81--0.51), white matter volume (SMD=-0.51, 95%CI: -0.64--0.38), gray matter volume (SMD=-0.60, 95%CI: -0.93--0.28), Cerebellum volume (SMD=-0.45, 95%CI: -0.64--0.25), hippocampus volume (SMD=-0.48, 95%CI: -0.73--0.24) and corpus callosum volume (SMD=-0.43, 95%CI: -0.63--0.24) in the experimental group were significantly reduced, respectively. Meta-analysis of whole-brain and white matter volume using funnel plot tests did not find publication bias.Conclusion: Very preterm birth and very low birth weight have a significant negative effect on volumes of whole brain and specific region of brain in adolescents.
AbstractObjective: To assess the effectiveness and safety of vitaminA(VitA) for Preventing bronchopulmonary dysplasia (BPD) in preterm infants systematically. Methods: The PubMed, EMBASE,Cochrane LibraryClinical controlled trial database, Chinese Journals full-text Database (CNKI), Chinese Technological Journals Database (VIP) and Wan Fang Digital Journal Full-text Database were searched for randomised control trials (RCTS) on whether VitA could prevent BPD in preterm infants (<37 weeks of gestational age) . The retrieval time was from inception to Oct.2017,at the same time, MESH and free words were used to retrieve. The primary outcome was the incidence of BPD and mortality before discharge; secondary outcomes included the rate of retinopathy of prematurity (ROP), patent ductus arteriosus (PDA) and intraventricular hemorrhage (IVH), hospitalization time, oxygen time, mechanical ventilation time, adverse reactions during the treatment. Bias risk was assessed based on the Cochrane handbook for systemic reviews. Revman 5.3 software was used for meta-analysis.Results: A total of 6 RCTS (both in English) including 1170 preterm infants met the inclusion criteria. The way of administration was intramuscular injection except a study's way was gastric tube. 6 articles all described the method of generating random sequences, all of them referred to allocation concealment, no ending data loss or selective report results, and 5 trials used the double blind method. Meta-analysis showed that the incidence of BPD in VitA group was lower than that in control group ［OR=0.67, 95%CI: 0.52~0.87 and P=0.003］. There was no significant difference in the mortality before discharge［OR=1.05，95％CI：0.77~1.44，P=0.76］, as well as in the secondary outcomes, including the incidences of ROP, PDA, IVH, adverse reactions, hospitalization time, mechanical ventilation time, and oxygen time, between the two groups. Conclusion: VitA can reduce the incidence of BPD.
AbstractObjective: To investigate the incidence and clinical characteristics of Clostridium difficile infection (CDI) in children with antibiotic-associated diarrhea (AAD), and to provide evidence for the diagnosis and treatment of antibiotic-related CDI. Methods: Inpatient clinical data of antibiotic-associated diarrhea from June 1st 2016 to October 1st 2017 were collected in Children's Hospital of Fudan University. Toxin A/B were tested and Clostridium difficile (CD ) was cultured. Neonates, incomplete information such as lack of fecal conventional bacteria culture and virus detection and the repeated cases were excluded . Toxin A/B(+) or the colonoscopy suggested pseudomembranous colitis cases were CDI group, the rest were in non-CDI group.Results: There were 150 patients with AAD. The incidence of CDI was 16.0%(24/150). The onset age of diarrhea was from one month ten days to 15 years 2 months old (median age 1.4 years old), among them boys were 103(68.7%) cases. ①Among 24 CDI patients, 22 children had acute diarrhea,1 had aggravated chronic diarrhea, because of Crohn's disease,1 had transitive diarrhea due to rectal polyp,11 (45.8%) had fever ,8(33.3%) had vomitus，2(8.3%) had abdominal pain,1 had abdominal distension ,1 had pseudomembranous colitis. ②There was no significant difference(P>0.05) in age of onset of diarrhea, gender, concomitant disease and antibiotic therapy two months before diarrhea between CDI group and non-CDI group. There was no statistic difference(P>0.05) in operations history, glucocorticoids and acid inhibitors therapy one month before suffering from diarrhea and lab index. The multiple factors Logistic models showed that clinical characteristics and routine laboratory test indicators had no statistically significant diffierence (P>0.05) between CDI group and non-CDI group.③The main treatments of AAD were to discontinue the broad-spectrum antibiotics, probiotics adjuvant therapy. If symptoms of CDI patients were not improved, metronidazole was the first choice.Failure to respond to metronidazole within 5-7 days should chang to vancomycin (oral) at standard dosing. The patients with symptomatic treatments of diarrhea were relieved in all CDI group，117 were improved in non-CDI group, 9 died of non-diarrhea. Only clinical performance and routine laboratory test indicators can't distinguish CDI and non-CDI among children AAD.Conclusion: The incidence of CDI of AAD patients was 16.0%. Fever and vomiting were the most common manifestations and prognosis was good. Only clinical performance and routine laboratory test indicators can't distinguish CDI and non-CDI among children AAD.
AbstractObjective: To analyze and discuss the risk factors of biliary atresia, so as to provide the possible clues for the early differential diagnosis and the study of its etiology and pathogenesis.Methods: Questionnaire of biliary atresia risk factors was designed, including a total of 22 factors.Patients with unexplained jaundice admitted to Children's Hospital Of Chongqing Medical University from July 2016 to June 2017 were enrolled.The clinical data of all patients were collected in September 2017, the biliary atresia group and the control group were determined and matched by sex and age. Using a matched pair study, the relevant risk factors were analyzed in both one-way ANOVA analysis and multivariate Logistic regression analysis. Results: A total of 146 cases were surveyed by questionnaires, and the questionnaires were returned to meet the inclusion and exclusion criteria of biliary atresia group and control group. There were 96 biliary atresia cases and 42 control cases. 42 cases of biliary atresia were matched to control group ,84 cases entered the analysis in the end.In one-way ANOVA analysis, pregnant four-dimensional color Doppler ultrasound, birth weight, adverse events at birth, stool color becoming shallow, trace elements supplement during pregnancy, folic acid supplement during pregnancy, pregnancy complication were statistically significantly different. In multivariate Logistic regression analysis, pregnancy complications and stool color becoming shallow were independent risk factors of biliary atresia (OR=21.27，95%CI：3.50~129.21，P=0.03；OR=6.70，95%CI：1.73~26.02，P=0.01); low birth weight was the independent protective factor of biliary atresia (OR=0.05，95%CI：0.00~0.67，P=0.03). Conclusion: Pregnancy complications of mother (gestational diabetes mellitus, hypertension, intrahepatic cholestasis) and color stool becoming shallow are the risk factors of biliary atresia,they have certain guiding values for early differential diagnosis.of biliary atresia.
AbstractObjective: To explore the clinical and genetic features of CDKL5 disorder.Methods: Using the next genomic sequencing technology to detect 7 cases of unexplained early infantile epileptic encephalopathy diagnosed by Xiangya Hospital of Central South University from the year 2011 to 2017,the clinical and molecular features of 7 cases with CDKL5 mutations were summarized ,and the related literatures were reviewed. Results: Five females and two males were diagnosed as CDKL5 disorder.Before the genetic tests,four patients were diagnosed as west syndrome and the other three patients were diagnosed as unclassified early onset epilepsy encephalopathy. The median age of seizure onset was 2 months,and the epilepsy started with partial seizure or tonic seizure,then turned into spasm or myoclonus.The EEG findings in five patients indicated hypsarrhythmia.Seven mutations of CDKL5 were found in seven patients,including three missenses mutations(c.464G>A / p．G134P、c.58G>C / p．G20R、c.464G>A / p．G155D),four frameshift mutations(c.1110del C / p．E370fs、c.160-163del / p．K54fs,c.278dupA / p．E93fs,heterozygous large deletion of the exon 3-8).All the mutations were not found in the parents.The median follow-up time was 36 months,the epileptic seizure of all patients could not be controlled.The number of antiepileptic drugs used ranged from 1 to 8.Four cases underwent the ACTH therapy but didn't get remarkable progress, five cases received ketogenic diet and only two of them achieved a little.All the patients had psychomotor develpoment retardation.Conclusion: CDKL5 disorder is a newly defined central nervous system syndrome caused by CDKL5 mutation,patients with different mutations have different phenotypes and prognosis.
AbstractObjective: To summarize the clinical and genotypic features of 3 childhood interstitial lung disease（chILD）caused by SFTPC mutations with reduced penetrance. Methods: The clinical and genetic data of the 3 chILD patients caused by SFTPC mutations with reduced penetrance were analyzed. Literature on genotypes and phenotypes of chILD caused by SFTPC mutations were reviewed. Results: The three cases were full-term newborns with uneventfully delivery. They developed cough, tachypnea, cyanosis and continuous oxygen dependence during 2-15 months of age. One patient's brother has chILD consistent with SP-C (surfactant protein C) dysfunction. Pathogenic heterozygous missense mutations in SFTPC were detected in all 3 causes (c.218T>C, p.I73T, the most common SFTPC mutation reported, in 2 girls; c.314A>G, p.D105G, in a boy). There are healthy carriers in all 3 families which were found by pedigree validation. They are treated with mechanical ventilation, exogenous surfactant, steroid, and hydroxychloroquine. Conditions of 2 cases improved and 1 died respectively. There are 10 cases caused by SFTPC mutations with reduced penetrance reported in 7 literature with complete details. Altogether with the 3 cases in this study, chest CT of all 13 cases showed diffuse ground-glass changes and the onset ranged from birth to 11 years old. As for the outcomes, 2 cases died, 11 survived with chILD, and conditions of 9 improved. Conclusion: Reduced penetrance of chILD caused by SFTPC mutations also exists in Chinese Han population. Understanding of the clinical characteristics and genotypes of chILD contributes to early diagnosis, intervention, prognosis evaluation and genetic counseling.
AbstractObjective: To summarize and review the clinical data of a case with systemic lupus erythematosus(SLE) caused by germline mutation in NRAS gene so as to improve the knowledge in spectrum of phenotype. Methods: Clinical data of a case with SLE were summarized, including clinical manifestations, laboratory findings and family investigation. The next generation sequencing was used to screen all exons of genomics in proband and her parents. Significant variants detected by next generation sequencing were confirmed by conventional Sanger sequencing and segregation analysis was performed using parental DNA and her brother samples.Results: The proband, 2.6-year-old boy, presented with recurrent fever and thrombocytopenia after age of 1 year. She often had rash, left knee joint swelling and pain, and mild proteinuria since the age of 2.6 years. At the same time, laboratory tests showed that ANA, anti-dsDNA antibody and other autoantibodies were positive. The patient had no abnormal facial features and no abnormalities in other organs. Whole exon sequencing was performed in core family, including proband and her parents. Heterozygous c.38G>A (p.G13D) mutation in NRAS gene was detected in proband, which was not carried by his parents. It was a de novo mutation.The mutation was confirmed by Sanger sequencing in core family. Conclusion: This study showed that a patient with germline mutation in NRAS gene could cause presenting with only SLE phenotype, and further extended the spectrum phenotype of mutations in NRAS gene.
AbstractObjective: To detect ZFPM2 mutations in conotruncal abnormality (CTA) patients by Sanger sequencing and study its pathogenicity and related molecular mechanism in CTA.Methods: Sanger sequencing was performed to detect ZFPM2 mutations in myocardial tissues of non-syndromic CTA patients in Children's Hospital of Fudan University. Plasmids were constructed, including expression constructs encoding GATA4, ZFPM2 wild type(WT) or variant and a reporter construct containing ANF promoter. The co-immunoprecipitation (Co-IP) and luciferase reporter gene assays were performed to study the change of interaction between ZFPM2 and GATA4 or the effect on ANF promoter activity in ZFPM2 WT or mutant. Injection of mRNA was performed to determine the effect of hZFPM2 mutant on cardiac phenotypes of zebrafish.Results: 107 non-syndromic CTA patients including 64 males and 43 females, aged from 1 to 168 months (median 8 months), were sequenced. Among them, there were 39 tetralogy of Fallot (TOF), 29 TOF with patent foramen ovale (PFO), 6 TOF with atrial septal defect (ASD), 8 TOF with PFO and patent ductus arteriosus (PDA), and other 25 multiple phenotypes, such as atrioventricular septal defects (AVSD) and left superior vena cava (LSVC). A heterozygous missense variant c.397A>G, p.M133V (rs77117583) was found in a CTA patient with TOF and PFO, which was located in the exon 4 and N-terminal transcriptional repression domain with 0.3‰ mutation frequency in Asian population. The Co-IP and luciferase reporter assays in HEK293T and HL-1 cells revealed that the M133V variant protein enhanced significantly the interaction between ZFPM2 with GATA4. And M133V mutant displayed a higher repression for GATA4-mediated transcriptional activation of ANF. Injection of hZFPM2 mRNA in zebrafish embryos demonstrated that the rate of cardiac abnormalities in M133V group(27.5%) was higher than control group without injection(11.3%) and WT group(15.0%), P＜0.05. The cardiac malformations were observed, including abnormal looping and the ventricle and atrium displayed the complete mirror image of normal.Conclusion: A low frequency variant of ZFPM2 c.397A>G, p.M133V was detected in a CTA patient and it was involved in the pathogenesis of CTA by affecting the combination with GATA4, and GATA4 transcriptional regulation.
AbstractObjective: To construct lentivirus-mediated shRNA targeting β-catenin to infect DOAY cells, to observe the expression of β-catenin and the biological behavior of cells. Methods: 3 kinds of shRNA targeting β-catenin were designed and inserted into the pCH-CMV-MCS-EF1-copGFP, the recombinant pCHD-shRNA was obtained. The pCHD-shRNA was transfected into HEK293T cells, and the lentivirus-shRNA was obtained. DOAY cells were infected by lentivirus-shRNA and divided into 5 groups: Lentivirus-shRNA 1group (LV-shRNA 1), Lentivirus-shRNA 2 group (LV-shRNA 2), Lentivirus-shRNA 3 group (LV-shRNA 3), Lentivirus-NC group (LV-NC) and blank group (Blank). After 72 h, the expression of green fluorescent protein (GFP) was observed, the expression of β-catenin was detected by reverse transcription PCR and Western Blot, the β-catenin gene stable suppression cells were screened out by puromycin. Effects of silencing β-catenin expression on cell biological behavior were observed. There were 3 groups: LV-shRNA 1, LV-NC and Blank. The proliferation and apoptosis of cells were detected by MTT assay and flow cytometry, the invasion and migration of the cells were detected by Transwell and cell scratch test. Results: The lentivirus-mediated shRNA targeting β-catenin was constructed successfully. 72 h after injection, the expression of GFP was not found in the blank group, the infection efficiency of LV-shRNA 1, LV-shRNA 2, LV-shRNA 3 and LV-NC was more than 89% (P＞0.05). The expression of β-catenin mRNA (0.24±0.09) and protein (0.15±0.07) in LV-shRNA 1 group was significantly lower than that in the other 4 groups, and the difference was statistically significant (P<0.05). Compared with LV-NC and blank group, the OD (570 nm) values of LV-shRNA 1 group at 24, 36, 48, 72 h were significantly decreased, the total apoptosis rate (31.28%) was significantly increased, the numbers of crossed cells (8.41±21.27) and migration distance (497.33±45.28) μm were significantly reduced, the difference was statistically significant (P<0.05).Conclusion: Lentivirus-mediated shRNA targeting β-catenin was successfully constructed. LV-shRNA 1 could significantly down-regulate the expression of β-catenin, significantly inhibit the proliferation of DOAY cells, reduce their invasion and migration ability, promote their apoptosis.
