Objective To explore the effects of obesity on the peak level of luteinizing hormone (LH) in the gonadotropin-releasing hormone (GnRH) agonist test and obesity-related hormones in girls with central precocious puberty (CPP). Methods Three hundred and thirty-three girls with CPP who underwent the GnRH agonist test between 2012 and 2014 were classified into three groups: normal weight (n=123), overweight (n=108), and obesity (n=102), according to body mass index (BMI). The sexual development indices were compared between the three groups. Twenty girls were randomly selected from each group for evaluation of the serum levels of leptin, sex hormone binding globulin (SHBG), neurokinin B, and kisspeptin. The correlation of BMI with the levels of various hormones was assessed using Pearson correlation analysis. Results There was no significant difference in mean age at diagnosis between the three groups; however, the bone age was significantly higher in the overweight and obesity groups than in the normal weight group (P< 0.05). The peak level of LH in the GnRH agonist test and SHBG level in the normal weight group were significantly higher than those in the overweight and the obesity groups, while the serum levels of leptin and neurokinin B were significantly lower in the normal weight group than in the overweight and the obesity groups (P< 0.05). BMI was negatively correlated with the peak level of LH in the GnRH agonist test and SHBG level (P< 0.05), and positively correlated with the levels of leptin and neurokinin B (P< 0.05). Conclusions The effects of BMI on the result of the GnRH agonist test and levels of obesity-related hormones should be taken into account in girls with precocious puberty.

Objective To analyze mutation types, clinical features, and treatment outcomes of cobalamin C (cblC) type combined methylmalonic aciduria and homocystinuria (MMA-HC) and to investigate the relationship of genotypes with clinical phenotypes and outcomes. Methods The clinical data of 16 Chinese children diagnosed with cblC type MMA-HC by gene analysis were retrospectively analyzed. According to the onset age, the patients were classified into early onset (≤1 year) and late onset (> 1 year). According to the clinical phenotype, the patients were classified into mild, moderate, and severe groups. All the patients were treated with vitamin B12 (cyanocobalamin) or hydroxocobalamin, betaine, folate, vitamin B6, and L-carnitine. Results Fifteen patients belonged to the early onset type, including 11 in the severe group and 4 in the moderate group. The remaining one belonged to the late onset type. Seven reported mutations and two novel mutations (c.445_446delTG and c.349G> c) were detected. The c.609G> A and c.658_660delAAG were the most common mutations detected in 13 (81%) out of 16 patients. The genotype caused by compound heterozygous mutations of these two alleles (c.609 G> A/c.658_660delAAG) was the most common in the patients, detected in 4 (25%) out of 16 patients. Patients with this genotype had severe microcephaly and eye diseases and these clinical manifestations were not improved after the treatment. The patient with late-onset cblC type MMA-HC had normal clinical phenotypes after treatment. In the 15 early onset patients, the more severe the clinical phenotype, the worse the treatment outcome. Conclusions The cblC type MMA-HC mainly manifests as early onset in China and c.609G > A and c.658_660delAAG are the most common mutations causing this disease. The clinical phenotypes are associated with the outcomes in children with cblC type MMA-HC.

Objective To study the molecular genetic mechanism and genetic diagnosis of pyruvate dehydrogenase complex deficiency (PHD), and to provide a basis for genetic counseling and prenatal genetic diagnosis of PHD. Methods Polymerase chain reaction (PCR) was performed to amplify the 11 exons and exon junction of the PDHA1 gene from a child who was diagnosed with PHD based on clinical characteristics and laboratory examination results. The PCR products were sequenced to determine the mutation. An analysis of amino acid conservation and prediction of protein secondary and tertiary structure were performed using bioinformatic approaches to identify the pathogenicity of the novel mutation. Results One novel duplication mutation, c.1111_1158dup48bp, was found in the exon 11 of the PDHA1 gene of the patient. No c.1111_1158dup48bp mutation was detected in the sequencing results from 50 normal controls. The results of protein secondary and tertiary structure prediction showed that the novel mutation c.1111 _1158dup48bp led to the duplication of 16 amino acids residues, serine371 to phenylalanine386, which induced a substantial change in protein secondary and tertiary structure. The conformational change was not detected in the normal controls. Conclusions The novel duplication mutation c.1111_1158dup48bp in the PDHA1 gene is not due to gene polymorphisms but a possible novel pathogenic mutation for PHD.

Objective To study the clinical characteristics of children with an initial onset of IgA nephropathy with nephrotic syndrome and compare them with children with primary nephrotic syndrome, in order to provide a theoretical basis for the differential diagnosis of the two diseases. Methods Fifty children diagnosed with an initial onset of IgA nephropathy with nephrotic syndrome were included in this study. Seventy-two children diagnosed with an initial onset of primary nephrotic syndrome served as the control group. The clinical and laboratory examination characteristics were compared between the two groups. Results The IgA nephropathy group had significantly higher incidence rates of gross haematuria, microscopic haematuria, hypertension, acute kidney injury, low serum high-density lipoprotein cholesterol, anemia, low serum complement C4, steroid resistance, and nephritis-type nephrotic syndrome and a significantly lower incidence of elevated serum IgE compared with the control group (P< 0.05). There were significant differences in serum creatinine, serum uric acid, serum total cholesterol, serum high-density lipoprotein cholesterol, serum IgE, serum complement C4, and hemoglobin levels between the IgA nephropathy and the control groups (P< 0.05). The thresholds of serum IgE (< 131.2 IU/mL) and high-density lipoprotein cholesterol (< 1.35 mmol/L) were reference parameters in the differential diagnosis of IgA nephropathy with nephrotic syndrome and primary nephrotic syndrome. Conclusions Children with IgA nephropathy presenting nephrotic syndrome manifest mainly as nephritis type and steroid-resistant type in the clinical classification. Cinical manifestations accompanied by serum levels of high-density lipoprotein cholesterol and IgE are helpful for differential diagnosis of IgA nephropathy presenting nephrotic syndrome and primary nephrotic syndrome.

Objective To study the effects of hemoperfusion treatment on serum interleukin-17 (IL-17) and IL-23 levels in children with Henoch-Schönlein purpura (HSP). Methods Eighty-seven children who were diagnosed with HSP and who had received hemoperfusion treatment between January 2011 and December 2012 were enrolled. Twenty-seven sex- and age-matched healthy children were recruited as normal controls. Enzyme-linked immunosorbent assay (ELISA) was used to measure serum concentrations of IL-17 and IL-23. Results The serum IL-23 and IL-17 levels in the HSP group were significantly higher than in the control group (P< 0.05). After hemoperfusion treatment, the serum IL-23 and IL-17 levels in the HSP group were significantly reduced to the levels of the control group. Serum serum IL-17 level was positively correlated with serum IL-23 level (P< 0.05) in children with HSP. Conclusions Hemoperfusion treatment can reduce serum IL-23 and IL-17 levels in children with HSP, suggesting that the treatment may be effective for HSP.

Objective To study the diagnostic values of fractional exhaled nitric oxide (FeNO) for typical bronchial asthma and cough variant asthma in children, and to explore whether FeNO can be applied to differentiate typical bronchial asthma from cough variant asthma in children. Methods A total of 150 children who were newly diagnosed with typical bronchial asthma between June 2012 and June 2014, as well as 120 children who were newly diagnosed with cough variant asthma during the same period, were selected as subjects. FeNO measurement, spirometry, and methacholine provocation test were performed for both groups. Meanwhile, 150 healthy children were selected as the control group, and their FeNO was measured. The diagnostic values of FeNO for typical bronchial asthma and cough variant asthma were analyzed using the receiver operating characteristic curve. Results The FeNO values in the typical bronchial asthma and cough variant asthma groups were significantly higher than in the control group (P< 0.01), and the FeNO value in the typical bronchial asthma group was significantly higher than in the cough variant asthma group (P< 0.01). FEV1/FVC%, FEV1%pred, and PD20 were significantly lower in the typical bronchial asthma group than in the cough variant asthma group (P< 0.01). The optimal cut-off value of FeNO was 19.5 ppb for the diagnosis of typical bronchial asthma, with a sensitivity of 83.3% and a specificity of 86.7%; the optimal cut-off value of FeNO was 15.5 ppb for the diagnosis of cough variant asthma, with a sensitivity of 67.5% and a specificity of 78.0%; the optimal cut-off value of FeNO was 28.5 ppb for the differentiation between typical bronchial asthma and cough variant asthma, with a sensitivity of 60.7% and a specificity of 82.5%. Conclusions Measurenment of FeNO may be useful in the diagnosis and differential diagnosis of typical bronchial asthma and cough variant asthma.

Objective To study the association between serum-specific immunoglobulin E (sIgE) allergens and asthma in children. Methods The serum sIgE allergens were determined using Western blot in 2 239 children aged 1-14 years, consisting of 1 415 children with asthma alone and 824 children with non-allergic diseases between December 2004 and April 2013. The case-control models of asthma alone and non-allergic diseases were established. The association between allergens and asthma was investigated using multivariate logistic regression analysis. Results In the 2 239 children, 1 028 children (45.91%) were serum sIgE-positive, and the allergen with the highest positive rate was house-dust mite (15.68%), followed by house dust (14.29%) and moulds (13.40%). The results of the case-control analysis showed that house-dust mite, moulds, house dust, and cashew nut/peanut/soybean were significantly associated with the development of asthma. House dust was associated with the development of asthma in the 1-2 years old group (P< 0.05). House dust and house-dust mite as allergens were identified as the risk factors for the development of asthma in the 3-14 years old group (P< 0.05). In the 6-14 years old group, moulds as allergens were identified as the risk factors for the development of asthma (P< 0.05). House dust and house-dust mite as allergens increased the risk of asthma in boys and girls, while moulds and cashew nuts/peanuts/soybeans as allergens increased the risk of asthma in boys. Conclusions House-dust mite, house dust, and moulds are the most common allergens in children with asthma, and they are closely associated with the development of asthma.

Objective To study the association of single nucleotide polymorphisms (SNP) (rs2393903 and rs10995251) in ZNF365 gene with bronchial asthma and its clinical characteristics in Han Chinese children in Hubei, China. Methods A total of 221 children with bronchial asthma and 243 normal children, all of whom were from Hubei, were recruited to carry out a case-control study. The genotype and allele frequencies of two SNPs in ZNF365 gene were determined using the polymerase chain reaction-restriction fragment length polymorphism technique. Results There were no significant differences in the distribution of three genotypes (GG, GA, AA) and allele frequency in SNP rs2393903 between the asthma and control groups (P> 0.05). However, there were significant differences in the distribution of three genotypes (CC, CT, TT) and allele frequency in SNP rs10995251 between the asthma and control groups (P< 0.05); C allele was a risk factor (OR=1.380). The asthmatic children with CC genotype of SNP rs10995251 had a significantly higher serum level of total immunoglobulin E (IgE) than those with TT genotype (P< 0.05). Conclusions The SNP rs10995251 in ZNF365 gene is associated with the susceptibility to bronchial asthma in children in Hubei, China, and the SNP may affect the level of serum IgE in these children.

Objective To study the efficacy and safety of Chinese Childhood Leukemia Group ALL 2008 (CCLG-ALL2008) protocol combined with tyrosine kinase inhibitor (TKI, imatinib) for the treatment of Philadelphia chromosome-positive (Ph⁺) acute lymphoblastic leukemia (ALL) in children. Methods The clinical data of 53 patients aged less than 15 years when first diagnosed with Ph⁺ ALL between October 2008 and December 2013 were retrospectively analyzed. The patients were assigned to two groups: HR (n=26) and HR+TKI (n=27). The HR group was treated with CCLG-ALL2008 protocol (for high-risk patients). The HR+TKI group was treated with imatinib in combination with CCLG-ALL2008 protocol (for high-risk patients). Results The complete remission rate and chemotherapy induction-related mortality rate in the TKI+HR and HR groups were 100% vs 75% and 0 vs 15%, respectively. The 3-year event-free survival (EFS) rate in the HR group was (6±5)%; the 5-year EFS rate of the TKI+HR group was (52±11)%. Compared with the HR group, the TKI+HR group had no increase in the toxic responses to chemotherapy and had a decrease in the infection rate during the induction period. Conclusions Application of imatinib significantly improves the clinical efficacy in children with Ph⁺ ALL and has good safety.

Objective To measure the expression of lymphocyte function-associated antigen-3 (CD58) in childhood B-lineage acute lymphoblastic leukemia (B-ALL) and to explore the feasibility of CD58 as an indicator for minimal residual disease (MRD) detection in childhood B-ALL. Methods Eighty-seven children diagnosed with B-ALL between January 2014 and September 2014 were enrolled, and 20 hospitalized children who had no tumor or blood disease and had normal bone marrow cell morphology served as the control group. The expression features of CD58 in bone marrow samples from the two groups (at diagnosis, on day 15 of induction chemotherapy) were analyzed by four-color flow cytometry (FCM). Quantitative real-time polymerase chain reaction (qRT-PCR) and FCM were used to detect MRD in B-ALL patients on day 33 of induction chemotherapy. Results The mean fluorescence intensity of CD58 expression in the 87 B-ALL cases (91±33) was significantly higher than that in the 20 controls (14±6) (P< 0.01); CD58 was over-expressed in 44 of the B-ALL cases. In the B-ALL children, the expression of CD58 on day 15 of induction chemotherapy (105±22) was not significantly different from that at diagnosis (107±26) (P> 0.05). In the 44 B-ALL patients with CD58 over-expression, FCM showed 9 MRD(+) cases and 35 MRD(-) cases, while qRT-PCR showed 11 MRD(+) cases and 33 MRD(-) cases; 42 cases (95%) showed consistent results of the two tests, so there was no significant difference between the two methods in detecting MRD (P> 0.05). Conclusions CD58 is over-expressed and stable in children with B-ALL, and it can be considered as an indicator for MRD detection in childhood B-ALL.

Objective To study the risk factors for moderate and severe iron deficiency anemia (IDA) in infants aged 6-12 months, and to preliminarily investigate the effects of IDA on the neuromotor development and temperament characteristics of infants. Methods A total of 326 infants aged 6-12 months with IDA were classified into three groups: mild IDA (n=176), moderate IDA (n=111), and severe IDA (n=39) according to the severity of anemia. The risk factors for moderate or severe IDA were investigated by multivariate logistic regression analysis. Three hundred and forty-six infants without IDA who showed matched age, sex, and other backgrounds were selected as the control group. The Gesell Development Diagnosis Scale was used to evaluate children's mental development. The Temperament Scale for infants was used for evaluating children's temperament. Results The univariate analysis showed that the severity of IDA was associated with sex, birth weight, gestational age, multiple birth, maternal anemia during pregnancy, and mother's lack of knowledge about IDA (P< 0.05). Setting the mild IDA group as control, the multivariate logistic regression analysis showed that multiple birth, premature birth, low birth weight (< 2 500 g), maternal anemia during pregnancy, breast feeding, and mother's lack of knowledge about IDA were the risk factors for severe IDA (OR> 1;P< 0.05); premature birth, breast feeding, and mixed feeding were the risk factors for moderate IDA (OR> 1; P< 0.05). The IDA group had significantly lower scores in Gesell general development quotient, gross motor, adaptive behavior, and fine motor than the control group (P< 0.05). The IDA group had higher percentages of children with difficulty and intermediate difficulty temperaments than the control group (P< 0.05). The IDA group had significantly higher scores in activity level, rhythmicity, adaptability, and perseverance than the control group (P< 0.05). Conclusions The severity of IDA is associated with premature birth, multiple birth, low birth weight, feeding pattern, maternal anemia during pregnancy and mother's lack of knowledge about IDA in infants aged 6-12 months. Infants with IDA have delayed neuromotor development and most of them have negative temperaments. More attention should be paid to mental and behavior problems for the infants. It is necessary to provide guidance for their parents in feeding and education.

Objective To examine serum 25-hydroxyvitamin D levels in children with attention deficit hyperactivity disorder (ADHD) and to explore the relationship between vitamin D level and ADHD. Methods Ninety-seven children with ADHD who were diagnosed according to DSM-V were selected as the ADHD group, including 46 cases of ADHD-I, 10 cases of ADHD-HI, and 41 cases of ADHD-C. Ninety-seven healthy children served as the control group. Serum levels of 25-hydroxyvitamin D were measured using electrochemiluminescence immunoassay. Results Mean serum levels of 25-hydroxyvitamin D in the ADHD group (17±7 ng/mL) were significantly lower than in the control group (23±8 ng/mL; P< 0.01). The serum levels of 25-hydroxyvitamin D in the three subtypes groups of ADHD (ADHD-I, ADHD-HI, and ADHD-C) were all lower than in the control group (P< 0.05). The rates of vitamin D insufficiency, deficiency or normal in the ADHD group were different from the control group (P< 0.01). The distributions of vitamin D levels in the three subtypes groups of ADHD were all different from the control group (P< 0.05). Conclusions Serum levels of 25-hydroxyvitamin D in children with ADHD are lower than in healthy children, suggesting vitamin D level might be related to ADHD.

Objective To investigate the growth rate of corpus callosum by cranial ultrasound in very low birth weight preterm infants and to provide a reference for early evaluation and improvement of brain development. Methods A total of 120 preterm infants under 33 weeks' gestation were recruited and divided into 26-29⁺⁶ weeks group (n=64) and 30-32⁺⁶ weeks group (n=56) according to the gestational age. The growth rate of corpus callosum was compared between the two groups. The correlation between the corpus callosum length and the cerebellar vermis length and the relationship of the growth rate of corpus callosum with clinical factors and the neuromotor development were analyzed. Results The growth rate of corpus callosum in preterm infants declined since 2 weeks after birth. Compared with the 30-32⁺⁶ weeks group, the 26-29⁺⁶ weeks group had a significantly lower growth rate of corpus callosum at 3-4 weeks after birth, at 5-6 weeks after birth, and from 7 weeks after birth to 40 weeks of corrected gestational age. There was a positive linear correlation between the corpus callosum length and the cerebellar vermis length. Small-for-gestational age infants had a low growth rate of corpus callosum at 2 weeks after birth. The 12 preterm infants with severe abnormal intellectual development had a lower growth rate of corpus callosum compared with the 108 preterm infants with non-severe abnormal intellectual development at 3-6 weeks after birth. The 5 preterm infants with severe abnormal motor development had a significantly lower growth rate of corpus callosum compared with the 115 preterm infants with non-severe abnormal motor development at 3-6 weeks after birth. Conclusions The decline of growth rate of corpus callosum in preterm infants at 2-6 weeks after birth can increase the risk of severe abnormal neuromotor development.

Objective To compare the differences of clinical efficacy between heated humidified high-flow nasal cannula (HHHFNC) ventilation and nasal continuous positive airway pressure (NCPAP) in the treatment of respiratory distress syndrome (RDS) in very low birth weight (VLBW) infants. Methods A total of 66 VLBW infants who were admitted to the neonatal intensive care unit were diagnosed with RDS, and they were randomly assigned to HHHFNC group and NCPAP group after receiving treatment with porcine pulmonary surfactant and conventional treatment. The changes in clinical symptoms and the incidence of complications were observed in the two groups. Results The HHHFN group had significantly earlier first milk feeding and full enteral feeding, significantly shorter oxygen exposure time and invasive ventilation time, and significantly lower incidences of second intubation within 7 days, nasal injury, air leak, and abdominal distention, as compared with the NCPAP group. Conclusions Compared with NCPAP, HHHFNC causes slighter injury and has better tolerability, and it can be considered as the first choice of noninvasive ventilation in the treatment of RDS in VLBW infants.

Objective To preliminarily investigate the long-term structural and functional injuries of mitochondria in rat brain caused by sepsis. Methods Wistar rats were randomly assigned into sepsis and control groups. A rat model of sepsis was prepared by an intraperitoneal injection of 10 mg/kg lipopolysaccharide (LPS) of gram-negative bacteria, and the survival assay was performed. Eight rats in the sepsis group were sacrificed at 12, 24, 48, or 72 hours after LPS injection, while rats in the control group were sacrificed after an intraperitoneal injection of an equal volume of normal saline. Mitochondria were extracted from rat brain tissue. Mitochondrial membrane potential (MMP) and mitochondrial swelling level were determined by flow cytometry, and the activities of electron transport chain complexes (I-V) were measured using enzyme assay kits. Hematoxylin-eosin (HE) staining and electron microscopy were used to observe morphological changes in brain tissue and mitochondria. Results The sepsis group had a significantly lower survival rate than the control group (P< 0.01). The MMP and activities of electron transport chain complexes (I-V) in the sepsis group, which were significantly lower than those in the control group (P< 0.05), were reduced to the lowest levels at 48 hours and partially recovered at 72 hours. The mitochondrial swelling level in the sepsis group, which was significantly higher than that in the control group (P< 0.05), increased to the peak level at 48 hours and partially recovered at 72 hours. Hematoxylin and Eosin staining revealed substantial damages in the structure of brain tissue, and electron microscopy showed mitochondrial swelling, and vacuolization in a few mitochondria. Conclusions In the rat model of LPS-induced sepsis, both structural and functional injuries are found in cerebral mitochondria, and achieve the peak levels probably at around 48 hours.

Objective To explore the effect of non-methylated cytosine-phosphate-guanine oligodeoxynucleotides (CpG-ODN) on serum transforming growth factor (TGF)-β and immune regulation in ovalbumin (OVA)-sensitized young mice. Methods Thirty female BALB/c mice (2-3 weeks old) were randomly divided into control, model, and CpG-ODN intervention groups. A young mouse model of food allergy was established by OVA sensitization. Normal saline of the same volume was used for replacement in the control group. The mice in the intervention group were intraperitoneally injected with CpG-ODN solution 1 hour before every OVA sensitization. Allergic symptoms were observed and scored for each group. The jejunal tissue was histopathologically examined with hematoxylin-eosin staining. Serum OVA-IgE level was measured using ELISA. Serum concentrations of interleukin (IL)-4, interferon (IFN)-γ, and TGF-β were determined by CBA. Results Allergic symptoms were observed in the model group and the jejunal tissue showed the pathological characteristics of type I allergic reaction. The allergic symptom scores in the model and CpG-ODN intervention groups were significantly higher than in the control group (P< 0.01). The serum levels of OVA-IgE, IL-4, and TGF-β were significantly higher in the model group than in the control and CpG-ODN intervention groups (P< 0.05). The CpG-ODN intervention group had significantly higher serum levels of OVA-IgE, IL-4, and TGF-β than the control group (P< 0.05). Compared with the control and CpG-ODN intervention groups, the model group had a significantly reduced IFN-γ level (P< 0.05). Conclusions The serum TGF-β level is increased in the young mouse model of OVA-sensitized food allergy and is involved in the allergy mechanism. Non-methylated CpG-ODN can reduce the serum TGF-β level in sensitized young mice and play an immunoregulatory role in food allergy.