Phenotypes and ATP7B gene variants in 316 children with Wilson disease / 中华儿科杂志

Objectives: To summarize the clinical phenotypes and the variation spectrum of ATP7B gene in Chinese children with Wilson's disease (WD) and to investigate their significance for early diagnosis. Methods: Retrospective analysis was performed on the clinical data of 316 children diagnosed as WD in Guangzhou Women and Children's Medical Center during the period from January 2010 to June 2021. The general situations, clinical manifestations, lab test results, imaging examinations, and ATP7B gene variant characteristics were collected. The patients were divided into asymptomatic WD group and symptomatic WD group based on the presence or absence of clinical symptoms at the time that WD diagnosis was made. The χ2 test, t test or Mann-Whitney U test were used to compare the differences between groups. Results: Among the 316 children with WD, 199 were males and 117 were females, with the age of 5.4 (4.0, 7.6) years at diagnosis; 261 cases (82.6%) were asymptomatic with the age of 4.9 (3.9, 6.4) years; whereas 55 cases (17.4%) were symptomatic with the age of 9.6 (7.3, 12.0) years. The main symptoms invloved liver, kidney, nervous system, or skin damage. Of all the patients, 95.9% (303/316) had abnormal liver function at diagnosis; 98.1% (310/316) had the serum ceruloplasmin lever lower than 200 mg/L; 97.7% (302/309) had 24-hour urine copper content exceeding 40 μg; only 7.4% (23/310) had positive corneal K-F rings, 8.2% (23/281) had abnormal MRI signals in the lenticular nucleus, and all of them had symptoms of damage in liver, kidney or nervous system. Compared with the group of symptomatic WD, asymptomatic group had higher levels of serum alanine aminotransferase and lower levels ceruloplasmin and 24-hour urine copper [(208±137) vs. (72±78) U/L, (55±47) vs. (69±48) mg/L, 103 (72, 153) vs. 492 (230, 1 432) μg; t=9.98, -1.98, Z=-4.89, all P<0.001]. Among the 314 patients completing genetic sequencing, a total of 107 mutations in ATP7B gene were detected, of which 10 are novel variants, and 3 cases (1.0%) had large heterozygous deletion (exons 10 to exon 11) in ATP7B gene. The percentage of missense mutation in asymptomatic WD children was significantly higher than that in symptomatic WD (81.5% (422/518) vs. 69.1% (76/110), χ²=8.47, P<0.05). WD patients carrying homozygous variant of c.2 333G>T had significantly low levels of ceruloplasmin than those not carrying this variant ((23±5) vs. (61±48) mg/L, t=-2.34, P<0.001). Conclusions: The elevation of serum ALT is an important clue for early diagnosis of WD in children, while serum ceruloplasmin and 24-hour urine copper content are specific markers for early diagnosis of WD. In order to confirm the diagnosis of WD, it is necessary to combine the Sanger sequencing with multiplex ligation-dependent probe amplification or other testing technologies.

Clinical features and StAR gene mutations in children with congenital lipoid adrenal hyperplasia / 中国当代儿科杂志

This article reported the clinical manifestations, steroid profiles and adrenal ultrasound findings in two unrelated Chinese girls with lipoid congenital adrenal hyperplasia (LCAH). Direct DNA sequencing and restriction fragment length polymorphism (RFLP) analysis were used to identify the mutations of steroidogenic acute regulatory protein (StAR) gene. The two patients with 46,XX karyotype, presented hyperpigmentation, growth retardation, and hyponatremia. Steroid profiles analysis revealed elevated plasma adrenocorticotrophic hormone levels, decreased or normal serum cortisol levels and low levels of androgens. Ultrasound examinations revealed that enlarged adrenals in patient 1 and normal adrenals in patient 2. Direct DNA sequencing of StAR gene showed a reported homozygous for c.772C>T(p.Q258X) in patient 1. Compound heterozygous for c.367G>A(p.E123K) and IVS4+2T>A (both novel mutations) were found in patient 2, inherited from her mother and father respectively. The amino acid of mutant position of the novel p.E123K was highly conserved in ten different species and was predicted to have impacts on the structure and function of StAR protein by the PolyPhen-2 prediction software. RFLP analysis revealed three bands (670, 423 and 247 bp) in patient 2 and her father and two bands (423 and 247 bp) in her mother and 50 controls. It is concluded that LCAH should be considered in girls with early onset of adrenal insufficiency and that steroid profiles, karyotype analysis, adrenal ultrasound and StAR gene analysis may be helpful for the definite diagnosis of LCAH.

Identification of a novel pathogenic mutation in PDHA1 gene for pyruvate dehydrogenase complex deficiency / 中国当代儿科杂志

<p><b>OBJECTIVE</b>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.</p><p><b>METHODS</b>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.</p><p><b>RESULTS</b>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.</p><p><b>CONCLUSIONS</b>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.</p>

Acupuncture combined with traction therapy for lumbar disc herniation: a systematic review / 中国针灸

To evaluate the efficacy and safety of acupuncture combined with traction therapy for lumbar disc herniation, providing the basis for future research strategies. Randomized control trials. (RCT) of acupuncture combined with traction therapy for lumber disc herniation at home and abroad from 2000 to 2013 were searched, analysis and evaluation of literature and strength of evidence were based on the principles and methods of Evidence-based Medicine. The total effective rate and curative rate were considered as primary outcome measures; pain improvement, quality of life, relapse rate and adverse effects were considered as secondary outcome measures. Seventeen RCTs were identified, Meta-analysis showed that (1) total effective rate and curative rate: acupuncture combined with traction therapy was better than single therapy (acupuncture or traction); (2) pain improvement: acupuncture combined with traction therapy was better than traction therapy; (3) relapse rate: current evidence could not support the conclusion that acupuncture combined with traction therapy was better than traction therapy. Acupuncture combined with traction therapy for lumbar disc herniation was effective. However, the included studies were with high risk of bias, important outcome measures such as quality of life, relapse rate and adverse effects were not found in most of the studies. Current evidence has not yet been able to fully reflect acupuncture combined with traction therapy for lumbar disc herniation is better than single therapy, so more RCTs of higher quality are needed to further confirm its efficacy and safety.

Protective effects of interleukin-10 and insulin-like growth factor-1 gene combination therapy on islet β cells of non-obese diabetic mice / 中华实用儿科临床杂志

Objective To investigate whether IL-10 gene combined with insulin-like growth factor-1 (IGF-1)gene transfer could attenuate pancreatic insulitis,increase the percentage of CD4 + CD25 + Foxp3 + regulatory T cells,and protect β cells from autoimmune destruction.Methods An adenoviral vector containing IL-10 gene (Ad-IL-10) or IGF-1 gene(Ad-IGF-1) was constructed separately.Forty female non-obese diabetic (NOD) mice were injected intraperitoneally with Ad-IL-10 and/or Ad-IGF-1,Ad-green fluorescent protein(GFP) and phosphate buffered saline(PBS)separately,repeated after 3 weeks.Blood glucose concentration was measured weekly.Serum insulin,cytokine production were tested by enzyme-linked immunosorbent assay.CD4 + CD25 + Foxp3 + Treg cells were determined by flow cytometry.Pancreatic histology was measured for determination of insulitis grades.Pancreatic insulin content and β-cell mass,proliferation were measured.Apoptosis was measured by using a terminal deoxynucleotidyl transferase dUTP nick end labeling assay.Results A significantly lower diabetes incidence (P ＜ 0.01) was observed in NOD mice treated with Ad-IL-10 and/or Ad-IGF-1,compared with mice treated with Ad-GFP or PBS alone,especially combined group.Lower insulitis score compared to control mice was found in Ad-IL-10 + Ad-IGF-1 group (all P ＜ 0.01).The serum level of TNF-α and IFN-γwere decreased and the level of IL-10 increased in combination therapy.The CD4 + CD25 +Foxp3 + cells was (7.17 ±0.38)％ in combined group,higher than that in the control groups.There was significantly less β-cell apoptosis(10.29 ±2.20)％ in combined group than that in other groups(all P ＜ 0.05).Conclusions Combination therapy with IL-10 and IGF-1 gene is able to increase the percentage of CD4 + CD25 + Foxp3 + regulatory T cells,reduce autoimmunity and increase pancreatic β-cell mass,indicating promising potential of these therapies as a new treatment strategy for diabetes mellitus.

Clinical and gene mutation analyses of three patients with ornithine carbamoyltransferase deficiency / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To analyze the clinical and genetic characteristics of three children with ornithine carbamoyltransferase deficiency(OTCD), and to provide a practical method for gene diagnosis and genetic counseling of the disease.</p><p><b>METHODS</b>All exons of the ornithine carbamoyltransferase (OTC) gene were screened by polymerase chain reaction-DNA direct sequencing in the three OTCD patients.</p><p><b>RESULTS</b>One patient firstly presented as vomiting at 6 month of age. A missense mutation of T262I was detected. His mother had the same mutation without any clinical symptoms. The second patient presented as restlessness, and had a missense mutation of R277W. Gene analysis of his parents was not available. The third patient presented as neonatal lethargy, harbored a missense mutation of I172M. His mother had the same mutation without any clinical symptoms.</p><p><b>CONCLUSION</b>Gene mutation analysis is a feasible way for diagnosing OTCD. Patients with I172M mutation present symptom early, while those with T262I and R277W mutations manifest symptoms later. Gene mutation analysis will be important for asymptomatic and prenatal diagnosis and genetic counseling.</p>

Neonatal diabetes mellitus: a clinical analysis of 13 cases / 中华儿科杂志

<p><b>OBJECTIVE</b>To study the clinical features of neonatal diabetes mellitus (NDM).</p><p><b>METHOD</b>Thirteen cases with NDM were seen in our department between Jul. 2004 and Sept. 2009. Their clinical features were reviewed retrospectively.</p><p><b>RESULTS</b>The average birth weight of the 13 cases was 2.30 kg. The median age at diagnosis was 2 months. The mean blood glucose at diagnosis was 22.2 mmol/L. Symptoms in 9 of 13 cases were exacerbated by infection and only 5 had typical symptoms of diabetes mellitus including polydipsia, polyuria, polyphagia and body weight loss. The common clinical findings included athrepsia, diuresis, and moderate dehydration. Ketoacidosis attacked 3 cases and 3 children had hypertriglyceridemia, meanwhile, 2 children had complications of blood clotting dysfunction and congenital cardiopathy, respectively. Autoantibody to insulin (IAA) was tested in 11 cases, all but one case was negative. Glycosylated hemoglobin was increased in 6 cases. Insulin treatment was started in all the 13 cases. The initial dose was 0.56-1.00 U/(kg × d), and the maximal dose was 1.35 U/(kg × d) depending on the variety of blood glucose. Blood glucose decreased significantly within 24 hours. Unfortunately, 1 case developed progressive blood glucose decline and recurrent hypoglycemia. Symptoms of the 3 cases who developed DKA were relieved 48 hours later, and their blood glucose was well under control. Among the 8 cases followed up, 4 had TNDM and 2 had PNDM. Unfortunately, 1 case died at the age of 3 months because insulin injection was stopped by the parents.</p><p><b>CONCLUSION</b>Early diagnosis and prompt management may lead to favorable prognosis. Blood glucose monitoring is a valuable method to avoid misdiagnosis and NDM should be differentiated from stress hyperglycemia, iatrogenic, or other causes of hyperglycemia.</p>

Persistent hyperinsulinemic hypoglycemia of infancy: clinical analysis of 12 cases / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To study the diagnosis and treatment of persistent hyperinsulinemic hypoglycemia of infancy.</p><p><b>METHODS</b>The clinical data of 12 infants with persistent hyperinsulinemic hypoglycemia were retrospectively reviewed.</p><p><b>RESULTS</b>Convulsion, cyanosis, lethargy, refusing milk sucking, irritability and sweating were common symptoms. The laboratory findings displayed persistent hypoglycemia and hyperinsulinism in all of the 12 infants. The glucagon test showed positive and no urine ketones were detected in all of the 12 infants. Seven infants were treated with oral diazoxide (5-15 mg/kg daily) and 4 infants showed effective to the therapy. One patient was given subtotal pancreatectomy and the blood glucose level was restored to normal after operation. Of the 12 infants, 6 presented psychomotor retardation in a follow-up of 2 months to 67 months, 3 had loss to follow-up and 3 were still in a follow-up.</p><p><b>CONCLUSIONS</b>The measurement of blood glucose, blood insulin and urinary ketons is helpful in the diagnosis of persistent hyperinsulinemic hypoglycemia of infancy. Diazoxide therapy is effective in some of patients.</p>

Complex glycerol kinase deficiency in three children / 中国当代儿科杂志

Glycerol kinase deficiency (GKD), a rare X-linked recessive disorder, is classified into two types: isolated and complex. Complex GKD is an Xp21 contiguous gene deletion involving the glycerol kinase locus together with the adrenal hypoplasia congenita (AHC) or Duchenne muscular dystrophy (DMD) loci or both. Its clinical features depend on the involved loci. GKD can be confirmed by an elevated urinary glycerol concentration tested by gas chromatography mass spectrometry (GC/MS). The three cases reported here were all male, presenting symptoms from neonatal period. The predominant clinical profile was characterized by hypoadrenocorticism, glyceroluria and Duchenne muscular dystrophy. After receiving a low fat diet and glucocorticoid replacement, they improved with relieved symptoms of hypoadrenocorticism. But they had significant developmental delays and myasthenia. In the follow-up two of them died of adrenal crisis.

Gene mutation analysis in four Chinese patients with multiple carboxylase deficiency / 中华儿科杂志

<p><b>OBJECTIVE</b>Multiple carboxylase deficiency (MCD) is an autosomal recessive disorder. MCD is characterized by skin rash, metabolic acidosis, vomiting and psychomotor retardation. Depending on deficiency of the enzyme, MCD includes two different forms, biotinidase deficiency (BTD, OMIM 253260) and holocarboxylase synthetase deficiency (HLCSD, OMIM 253270). In this study, we analyzed gene mutations of four Chinese MCD patients and to explore the mutation spectrum and possibility of a molecular diagnosis.</p><p><b>METHODS</b>All exons and their flanking introns of biotinidase gene and HLCS gene were screened by polymerase chain reaction combined with DNA direct sequencing in four Chinese MCD patients. Genomic DNA was extracted using a kit from the peripheral blood leukocytes of each patient. PCR amplification products were checked by 2% agarose gel electrophoresis and were subsequently sequenced with both the forward and reverse primers.</p><p><b>RESULTS</b>All patients showed mutations in HLCS gene, whereas no mutation was found in biotinidase gene, proving that all the four patients had HLCS deficiency. Four previously reported mutations in HLCS gene were detected (Y456C, R508W, D634N and 780delG). A missense mutation of 1522C > T in exon 11 of HLCS gene, which was a homozygotic mutation, was identified in patient 1; a mutation of 1522C > T in exon 11 combined with a mutation of 1367A > G in exon 9, which was a compound heterozygotic mutation, was identified in patient 2; a mutation of 1522C > T in exon 11 combined with a mutation of 1900G > A in exon 13, which was a compound heterozygotic mutation, was identified in patient 3; a mutation of 1522C > T in exon 11 combined with a mutation of 780delG in exon 7, which was a compound heterozygotic mutation, was identified in patient 4. All the parents were carriers of mutations. No additional carrier of this four mutations was identified from 50 samples of Chinese controls.</p><p><b>CONCLUSION</b>The 1522C > T (R508W) mutation probably represents a mutational hot-spot in Chinese HLCS deficiency patients while the 780delG mutation which was reported only in Japanese patients was found firstly in Chinese patients.</p>

Clinical and Laboratorial Assessment for Identifying Isolated Premature Thelarche and Central Precocious Puberty / 实用儿科临床杂志

3-8 years old group(group C2) with 50 cases] were detected.A gonadotropin releasing hormone analogue(GnRHa) stimulation test was performed in 140 girls with IPT.The 140 girls were divided into 3 groups:IPT group,CPP group,and peripheral precocious puberty group(PPP group).Kruskal-Wallis and Mann-Whitneg tests were performed on the data between every groups.Results For basal LH levels,there were significant diffe-rences between IPT1 group and group C1,among IPT2 group,CPP group and group C2(Pa0.05).For peak LH/FSH ratios,there was significant difference between IPT2 group and CPP group(P

Effects of power frequency magnetic field on gap junction intercellular communication of astrocytes / 中华劳动卫生职业病杂志

<p><b>OBJECTIVE</b>In order to explore if power frequency magnetic field (PFMF) can act as cancer promoter or be synergistic with phorbol 12-myristate 13-acetate (TPA) in cancer promotion, the effects of 50 Hz MF on gap junction intercellular communication (GJIC) of astrocytes were observed.</p><p><b>METHODS</b>Fluorescence redistribution after photobleaching (FRAP) was adopted to observe the recovery of fluorescence intensity in the bleached cells thus to estimate intercellular communication by gap junction. Comparative fluorescence intensity recovery rate (CFIRR) was as evaluation index. The effects of 50 Hz MF alone or with TPA on GJIC of astrocytes were studied.</p><p><b>RESULTS</b>After 3 ng/ml TPA treatment for 1 hour, M(d) of CFIRR was 4.53%/min, whereas that in the control group was 9.74%/min (H = 12.084, P < 0.005). After exposure to 0.8 and 1.6 mT magnetic field for 24 hours respectively, M(d) of CFIRR was 8.25%/min and 6.68%/min respectively, no significant difference from that of control (H = 32.617, P > 0.05). After exposure to 0.8 and 1.6 mT magnetic field for 23 hours then combined with 3 ng/ml TPA treatment for 1 hour, M(d) of CFIRR was 3.32%/min and 2.85%/min respectively, also no significant difference from that in the group treated with 3 ng/ml TPA alone (H = 2.589, P > 0.05).</p><p><b>CONCLUSION</b>50 Hz MF (within 0 - 1.6 mT) alone could not inhibit GJIC of astrocytes; with TPA, could not enhance the inhibition of TPA on GJIC of astrocytes. But with MF intensity increasing, the inhibition of MF on GJIC showed elevated tendency.</p>