Abstract:OBJECTIVE: To investigate the levels of CD4+CD25+CD127low regulatory T cells (Tregs) and the expression of Foxp3 gene in peripheral blood of children with aplastic anemia (AA) and to study their roles in the pathogenesis of AA. METHODS: Twenty-one children with chronic AA, 9 with acute AA and 15 healthy children were enrolled. The proportion of CD4+CD25+ CD127low Tregs in CD4+ T cells was evaluated by flow cytometric analysis. The level of Foxp3 mRNA was ascertained by RT-PCR. RESULTS: The percentage of peripheral blood CD4+T cells and CD4+CD25+ and CD4+CD25+CD127low Tregs in CD4+T cells in both the acute and chronic AA groups was significantly lower than that in the normal control group (P﹤0.05). The acute AA group had more decreased CD4+ T cells and CD4+CD25+ and CD4+CD25+CD127low Tregs percentage compared with the CAA group (P<0.05). The expression of Foxp3 mRNA in peripheral blood decreased obviously in the acute AA group (0.47±0.08%) compared with that in the normal control (0.71±0.12%) and the CAA groups (0.68±0.14%) (P<0.05). CONCLUSIONS: The low expression of Tregs and Foxp3 mRNA in peripheral blood may be involved in pathogenesis of AA. The more decreased Tregs and Foxp3 mRNA expression in acute AA than chronic AA suggests their possible roles in the assessment of the severity of AA.［Chin J Contemp Pediatr, 2010, 12 (4):241-243］

Abstract:OBJECTIVE: To investigate the efficacy of high dose chemotherapy combined with autologous peripheral blood stem cell transplantation (APBSCT) for the treatment of neural ectodermal solid tumor originated from neural crest in children. METHODS: Twenty-three children at a medium age of 5.8±3.5 years with neural ectodermal solid tumor originated from neural crest were enrolled. Of the 23 children, 20 with stage IV neuroblastoma (9 were in complete remission, 7 were in partial remission and 4 were in progressive disease), 2 with stage IV primitive neuroectodermal tumor (PNET) in complete remission, and 1 with retinoblastoma in partial remission. Before APBSCT the children received 8.0±4.3 courses of chemotherapy. During chemotherapy the autologous peripheral blood stem cells were harvested and the tumor excision was performed. Then APBSCT was performed. RESULTS: The reconstruction of the hematopoietic system was noted in 19 of 20 children with stage IV neuroblastoma 16.5±0.9 days after transplantation. A follow-up (median 15.8 months) was done in these children. The follow-up showed that the survival rate in children in complete remission before transplantation was 100%, 57% in those in partial remission, and none of children in progressive disease survived (P<0.05). The total survival rate was 67% in children with neuroblastoma. The child with retinoblastoma had complete remission in a 6-months follow-up. The tumors recurred in children with PNET 5 to 8 months after transplantation and all died within one year after transplantation. CONCLUSIONS: High dose chemotherapy combined with APBSCT can result in a good outcome in children with neural ectodermal solid tumor originated from neural crest in complete remission before transplantation and can improve the outcome in patients in partial remission before transplantation. However, the children with PNET, even in complete remission before transplantation, do not respond to the therapy,［Chin J Contemp Pediatr, 2010, 12 (4):244-247］

Abstract:OBJECTIVE: To assess the regional left ventricular systolic function of children with Kawasaki disease before and after treatment by vector myocardial strain and strain rate imaging (VSI) technology. METHODS: The regional left ventricular systolic function was assessed using VSI technology in 32 children with Kawasaki disease before treatment and one month after treatment and in 30 age-matched normal children. RESULTS: Nine segments of the left ventricular in the Kawasaki disease group before treatment had decreased longitudinal peak systolic strain rate (LSRs) compared with the normal control group. After treatment, the LSRs in 9 segments in the Kawasaki disease group increased, but 6 segments had decreased LSRs compared with the normal control group. The radial peak systolic strain rate (RSRs) of 8 segments in the Kawasaki disease group before treatment was lower than that in the control group. After treatment, only one segment had decreased RSRs compared with the control normal group and 5 segments had increased RSRs compared with that before treatment. The circumferential peak systolic velocity (CVs) of 6 segments in the Kawasaki disease group before treatment group was lower than that in the control normal group. After treatment, only one segment had decreased CVs in the Kawasaki disease group compared with the control normal group and 3 segments had increased CVs compared with that before treatment. CONCLUSIONS: The regional left ventricular systolic function in children with Kawasaki disease before and after treatment can be accurately assessed using VSI technology, which shows the clinical significance of this technology in assessment of treatment outcome in children with Kawasaki disease.［Chin J Contemp Pediatr, 2010, 12 (4):248-251］

Abstract:OBJECTIVE: To investigate the changes and the clinical significance of N-terminal pro-brain natriuretic peptide (NT-proBNP) and glycogen phosphorylase isoenzyme BB (GPBB) levels in neonates with asphyxia complicated by myocardial injury. METHODS: Sixty-four neonates with asphyxia (39 mild, 25 severe) were enrolled. Of the 64 neonates, 30 had myocardial injury and 34 did not develop myocardial injury. Twenty-five healthy neonates served as a control group. Plasma levels of NT-proBNP and GPBB were measured using ELISA. Myocardial enzymes and cardiac troponin I were stimultaneously measured, and electrocardiography and chest radiographs were obtained. RESULTS: The plasma levels of NT-proBNP and GPBB in neonates with myocardial injury were significantly higher than those in neonates without myocardial injury and in the control group (P<0.01). The neonates with severe asphyxia had significantly increased plasma NT-proBNP and GPBB concentrations compared to those with mild asphyxia and the control group (P<0.01). Spearman rank correlation analysis showed that plasma NT-proBNP level was positively correlated with plasma GPBB level in neonates with asphyxia. Plasma levels of NT-proBNP and GPBB were also positively correlated with plasma levels of CK-MB, CK and LDH (P<0.01). CONCLUSIONS: Both NT-proBNP and GPBB can be used as biomarkers of myocardial injury in neonates with asphyxia .The measurement of plasma NT-proBNP and GPBB levels was useful in early identification of myocardial injury and severity evaluation in neonates with asphyxia.［Chin J Contemp Pediatr, 2010, 12 (4):252-255］

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OBJECTIVE: To evaluate of the efficacy and safety of adjunctive levetiracetam (LEV) in children younger than 4 years with refractory epilepsy. METHODS: One hundred and twelve children at age of 4 months to 4 years with refractory epilepsy received LEV as adjunctive therapy. LEV was administered in two equal daily doses of 10 mg/kg. The dose was increased by 10 mg/kg every week up to the target dose (20-40 mg/kg). The efficacy and tolerability were evaluated. RESULTS: At an average follow-up period of 13 months (6-22 months), LEV administration was found to be effective in 43 children (38.4%) (responders showing more than a 50% decrease in seizure frequency) and 14 children (12.5%) became seizure-free. Fifty-three children (47.3%) did not respond to the treatment and 2 children (1.8%) worsened. The therapy-related adverse events were mild, including restlessness,. reduction in sleep time, night terrors, debility, somnolence, nausea and vomiting. The adverse events were either tolerable or resolved in time with dosage reduction in most of children, and only 3 cases required discontinuation. CONCLUSIONS: LEV as adjunctive therapy is effective and well-tolerated in children younger than 4 years with refractory epilepsy, suggesting that it represents a valid option for the treatment of refractory epilepsy in this age group.［Chin J Contemp Pediatr, 2010, 12 (4):256-258］

Abstract:OBJECTIVE: To study the changes in the hippocampal volume in the acute phase after febrile convulsions (FC) in children. METHODS: The brain MRI was performed on 30 children with FC (15 simple and 15 complex) and 30 normal children (control). The hippocampal volume for both sides was compared between groups. RESULTS: In the control group, the average volume of the right hippocampus was significantly larger than that of the left side (P<0.05). There was no significant difference in the hippocampal volume between the left and right sides in the FC group. The average volumes of both the left and right hippocampus in the FC group (2.03±0.25 cm3 and 2.18±0.21 cm3 respectively) were enlarged compared to the control group (1.90±0.24 cm3 and 1.97±0.20 cm3 respectively) (P<0.05). There was no significant difference in the hippocampal volume between the simple and the complex FC groups. CONCLUSIONS: The hippocampal volume is enlarged in the acute phase after FC in children. The hippocampal volume in children with simple FC is similar to that in children with complex FC in the acute stage.［Chin J Contemp Pediatr, 2010, 12 (4):259-261］

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OBJECTIVE: To investigate the relationship of the promoter of matrix metalloproteinase-9 (MMP-9) gene polymorphisms with the susceptibility and clinical features of Helicobacter pylori (H. pylori)-related chronic gastritis and duodenal ulcer in children. METHODS: One hundred children with chronic gastritis, 32 children with duodenal ulcer and 102 healthy children were enrolled．The promoter of MMP-9-1562C/T gene polymorphisms were genotyped by polymerase chain reaction restriction fragment length polymorphism (PCR-RFLP) and sequencing. MMP-9 mRNA expression in gastric mucosa was confirmed by reverse transcription polymerase chain reaction. RESULTS: The genotype distributions and allele frequencies of MMP-9-1562C/T gene polymorphisms were similar in gastric upper gastrointestinal disease and healthy subjects. The relative risk for H.pylori infection in C/C genetype carriers was 3.1 times as high as that in T allele (C/T+T/T) carriers in children with chronic gastritis. MMP-9-1562 C/T gene polymorphisms did not affect MMP-9 mRNA expression level. CONCLUSIONS: These data suggest that MMP-9-1562 C/T gene polymorphisms are not associated with susceptibility to chronic gastritis and duodenal ulcer in children. The C/C genotype of MMP-9-1562 C/T gene polymorphism might be associated with H.pylori infection.［Chin J Contemp Pediatr, 2010, 12 (4):262-266］

Abstract:OBJECTIVE: To investigate cagA, vacA and iceA genotypes of Helicobacter pylori (H. pylori) isolated from children suffering from gastric and duodenal diseases in Shanghai and to explore a possible genotype-phenotype correlation. METHODS: From May 2007 to January 2008, 59 children were confirmed with Hp infection by gastroscopy. Biopsied specimens were taken from the gastric antrum. cagA, vacA and iceA genes were determined by PCR. The histological changes in the gastric mucosa were evaluated. The levels of IFN-γ and IL-4 in the gastric mucosa were measured using ELISA. RESULTS: cagA, vacAs1/m1, vacAs1/m2, iceA1 and iceA2 were found in 65%, 19%, 40%, 63% and 19% of H. pylori strains, respectively. Both iceA1 and iceA2 were detected in 9% of strains. There were no statistical differences in the distribution of various genotypes between the children with chronic gastritis and peptic ulcer. No association was observed between the genotypes and the degree of inflammation of gastric mucosa. There were no significant differences in levels of IFN-γ and IL-4 in the gastric mucosa infected by different genotypes of H. pylori strains. CONCLUSIONS: cagA/vacAs1/m2/iceA1 may be the commonest genotype combination of H.pylori in children from Shanghai. That there was no association between H.pylori genotypes and clinical variables suggests the potential role of host and environment factors in the development of clinical diseases at a later life.［Chin J Contemp Pediatr, 2010, 12 (4):267-271］

Abstract:OBJECTIVE: To study the significance of plasma D-dimer and von Willebrand factor (vWF) and the therapeutic effect of compound glycyrrhizin in children with cytomegalovirus (CMV) hepatitis. METHODS: Twenty healthy children, 16 asymptomatic cases with CMV infection and 52 cases of CMV hepatitis (21 cholestatic and 31 non-cholestatic) were enrolled. The 52 children with CMV hepatitis were randomly administered with conventional treatment alone or conventional treatment plus compound glycyrrhizin treatment. Plasma D-dimer and vWF levels were measured before and after treatment. RESULTS: Plasma D-dimer and vWF levels in the CMV hepatitis group were markedly higher than those in the healthy control and asymptomatic CMV infection groups (P<0.01). The cholestatic hepatitis group had more increased plasma D-dimer and vWF levels compared with the non-cholestatic hepatitis group (P<0.01). Plasma D-dimer and vWF levels in the CMV hepatitis group were markedly reduced after conventional or compound glycyrrhizin treatment (P<0.01). Compound glycyrrhizin treatment decreased more significantly plasma D-dimer and vWF levels compared with the conventional treatment in children with CMV hepatitis (P<0.01). CONCLUSIONS: The detection of plasma D-dimer and vWF is useful in the early assessment of liver damage in children with CMV hepatitis. Compound glycyrrhizin can decrease obviously plasma D-dimer and vWF levels and might thus provide protective effects against liver damage.［Chin J Contemp Pediatr, 2010, 12 (4):272-274］

Abstract:OBJECTIVE: To examine serum levels of interleukin-13 (IL-13) and tumor necrosis factor alpha (TNF-α) in children with Mycoplasma pneumoniae (MP) pneumonia. METHODS: Eighty children with MP pneumonia complicated by wheezing or without (n=40 each), 40 children with pneumonia from non-MP infection and 40 healthy children were enrolled. Serum levels of IL-13 and TNF-α were measured using ELISA. RESULTS: The serum levels of IL-13 and TNF-α in the MP pneumonia group were significantly higher than those in non-MP pneumonia group and the healthy control group (P<0.01). The children with MP pneumonia complicated by wheezing had increased serum levels of IL-13 (214.6±67.2 ng/L vs 189.6±52.1 ng/L; P<0.01) and TNF-α(0.55±0.13 ng/mL vs 0.42±0.16 ng/mL; P<0.01)compared with those without wheezing. CONCLUSIONS: The increase in serum levels of IL-13 and TNF-α may play important roles in the pathogenesis of MP pneumonia and wheezing attack in children.［Chin J Contemp Pediatr, 2010, 12 (4):275-277］

Abstract:OBJECTIVE: To understand the prognosis of the children with urinary calculi associated with exposure to melamine-contaminated formula. METHODS: A follow-up study was performed in 47 out-patients from the Third Hospital of Peking University who were diagnosed with urinary calculi associated with exposure to melamine-contaminated formula. Urinary tract B-ultrasound and renal function examinations were done during the 1 to 6-month follow-up. RESULTS: By the 6th month of follow-up, spontaneous stone passage was found in 36 children (77%). The follow-up failed in four children. None of the patients had any complications. Spontaneous stone passage was not associated with the volume and the period exposed to melamine in formula, but was associated with the location of calculi. The time to spontaneous stone passage in boys appeared to be longer than in girls, but there were no statistical differences. CONCLUSIONS: Spontaneous stone passage can be found in most children and the prognosis of children with urinary calculi associated with exposure to melamine-contaminated formula is good.［Chin J Contemp Pediatr, 2010, 12 (4):278-279］

Abstract:OBJECTIVE: To study the expression of Toll-like receptor 4 (TLR4) in renal tubular epithelial cells exposed to high glucose and the effect of spironolactone on the TLR4 expression. METHODS: In vitro renal tubular epithelial cells (NRK-52E) were randomly exposed to DMEM culture solution with low glucose (5 mmol /L), high glucose (25 mmol/L) or 10-7 mol/L spironolactone plus 25 mmol/L glucose. Immunohistochemistry, RT-PCR and Western blot were used to determine TLR4 protein and mRNA expression. The levels of IL-6 and TNF-α in the cell culture supernatant were determined using ELISA. RESULTS: The expression of TLR4 mRNA in the high glucose group began to increase 6 hrs and remained at a higher level up to 24 hrs after exposure as compared with the low glucose group. The TLR4 mRNA expression in the spironolactone treatment group was significantly lower than that in the high glucose group, although it was higher than that in the low glucose group between 6 and 24 hrs after exposure. TLR4 protein expression increased significantly in the high glucose group 24 and 48 hrs after exposure compared with that in the low glucose group. The TLR4 protein expression in the spironolactone treatment group was lower than that in the high glucose group, but higher than that in the low glucose group. IL-6 and TNF-α expression in the supernatant from the NRK-52E cells in the high glucose groups increased significantly as compared with the low glucose group. The spironolactone treatment group had significantly reduced IL-6 and TNF-αexpression compared with the high glucose group. CONCLUSIONS: High glucose triggers an increase in the expression of TLR4 and inflammatory factors in NRK-52E cells. TLR4 may participate in the progress of diabetic nephropathy. Spironolactone can reduce expression of TLR4 and inflammatory factors, which might be attributed to one of the mechanisms of protection by spironolactone against diabetic nephropathy.［Chin J Contemp Pediatr, 2010, 12 (4):280-283］

Abstract:OBJECTIVE: Some research has shown that learning and memory function impairments in rats with hypothyroidism are associated with triiodothyronine (T3) deficiency in neurons. This study aimed to investigate the effects of L-T3 administration on learning and memory behaviors in neonatal mice with excitotoxic brain damage. METHODS: Seventy-one 5-day-old ICR neonatal mice were randomly assigned to five groups: controls that received intracerebral and intraperitoneal injections of phosphate buffered saline (PBS) (n=14); a group that received intracerebral injections of ibotenic acid (IA) and intraperitoneal injection of PBS (n=14); 3 groups that received intracerebral injections of IA and intraperitoneal injection of L-T3 at 0.2, 0.5, and 1 μg/kg, respectively (n=14-15). Intraperitoneal injections were done 1, 24, 48, 72 and 96 hrs after intracerebral injections. Learning and memory functions were evaluated by the Y-maze discrimination learning test on postnatal days 33-34. RESULTS: The learning and memory functions in the highest L-T3 dose group were significantly better than those in the IA, and the lower L-T3 dose groups, presenting with decreased number of trials to criterion ［15.8±4.5 vs 21.3±6.3 (IA group), 20.5±6.0 (0.2 μg/kg L-T3 group) or 21.0±6.5 (0.5 μg/kg L-T3 group); P<0.05］, and achieving a higher correct percentage ［91.4±9.5% vs 79.3±10.0% (IA group), 77.9±14.2% (0.2 μg/kg L-T3 group) or 80.7±12.2% (0.5μg/kg L-T3 group); P<0.05］. CONCLUSIONS: High-dose L-T3 (1 μg/kg) may improve learning and memory functions in mice following excitotoxic brain damage.［Chin J Contemp Pediatr, 2010, 12 (4):284-286］

Abstract:OBJECTIVE: To study the role of urotensionII in serum and bronchoalveolar lavage fluid (BALF) in the process of airway remodelling in asthmatic rats. METHODS: Thirty-two male Sprague-Dawley (SD) rats were randomly divided into normal control and 2-week, 4-week and 8-week asthmatic groups (OVA inhalation of 2, 4 and 8 weeks respectively). Rats were sensitized and challenged by OVA to establish a model of asthma. The bronchial wall thickness and the airway smooth muscle thickness were measured by image analysis system. The urotension-II contents in serum and BALF were determined using ELISA. RESULTS: The bronchial wall thickness and the airway smooth muscle thickness in the three asthmatic groups significantly increased compared with those in the normal control group (P<0.01). The urotension-II contents in serum and BALF in the three asthmatic groups also increased significantly compared with those in the normal control group (P<0.01). The urotension-II contents in serum and BALF in the 8-week asthmatic group were the highest, followed by the 4-week and the 2-week asthmatic groups (P<0.01). BALF urotension-II contents were positively correlated with the bronchial wall thickness and the airway smooth muscle thickness as well as serum U-II contents in the four groups. CONCLUSIONS: The urotension-II contents in serum and BALF in the process of airway remodeling increase in asthmatic rats. The changes in serum and BALF urotension-II contents may be associated with airway remodeling in asthmatic rats.［Chin J Contemp Pediatr, 2010, 12 (4):287-289］

Abstract:OBJECTIVE: To study the effect of astragaloside IV on the expression of cytokines in bone mesenchymal stem cells (MSCs) in rats. METHODS: MSCs were isolated from Wistar rats by the method of adhesive cultiration and clone, and then their biological activities were assessed using indirect immunofluorescence. Proliferation of MSCs stimulated with astragaloside IV was ascertained by the MTT method. Expression of cytokines was ascertained using RT-PCR in MSCs with astragaloside IV stimulation or not. RESULTS: MSCs were effectively isolated and purified in vitro, and had expression of many cytokines except IL-3, such as stem cell factor (SCF), thrombopoietin (TPO), granulocyte macrophage colony stimulating factor (GM-CSF) and transforming growth factor (TGF-β1). Astragaloside IV stimulation promoted MSCs proliferation, and 200 mg/mL astragaloside IV treatment produced a peak effect 72 hrs after culture. The SCF expression in MSCs stimulated with astragaloside IV increased significantly compared with that in MSCs without astragaloside IV stimulation. CONCLUSIONS: Astragaloside IV may promote MSCs proliferation and increase SCF secretion in vitro.［Chin J Contemp Pediatr, 2010, 12 (4):290-292］

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