OBJECTOVE: Early response to therapy is one of the most important prognostic factors in childhood acute lymphoblastic leukemia (ALL). This study aimed to assess the prognostic value of morphological assessment of bone marrow blasts during remission induction and determination of minimal residual disease (MRD) after remission induction. METHODS: From January 1998 to May 2003, 193 children with newly diagnosed ALL were enrolled on the ALL-XH-99 protocol. Blast cell count in the bone marrow was examined on day 19 of remission induction and by the completion of remission induction. MRD was measured with the flow cytometry. Event-free survival (EFS) was estimated by Kaplan-Meier analysis and the distributions of EFS were compared using the log-rank test. A Cox proportional hazards model was used to identify independent prognostic factors. RESULTS: The 4-year EFS was significantly worse in patients with ≥ 5% lymphoblasts in the bone marrow on day 19 as compared to those with <5% lymphoblasts on that date (42.59%±14.28% vs 74.24%±6.67%; P<0.01). The 4-year EFS was significantly worse in patients with any amount of lymphoblasts in the bone marrow on the remission date as compared to that of other patients with no morphologically identifiable blasts (63.47%±9.23% vs 76.41%±6.09%; P<0.05). The patients with MRD <0.01 had significantly better outcome than those with a level ≥ 0.01% (15-month EFS:94.44%±5.40% vs 23.81%±20.26%; P<0.01). CONCLUSIONS: Early treatment response as assessed by morphological examination or minimal residual leukemia determination by flow cytometry has important prognostic significance, and can be performed in a resource-poor patient population.［Chin J Contemp Pediatr, 2008, 11 (1):5-9］

OBJECTIVE: Caroli's syndrome is a rare autosomal recessive hereditary disease. Here a case of Caroli's syndrome associated with medullary sponge kidney was reported. The patient was a 2-years and 10 months-old boy. He presented with hepatosplenomegaly. Fever, abdominal pain or jaundice was not found. The imaging examination showed intrahepatic bile duct dilation, splenomegaly, medullary sponge kidney and nephrocalcinosis. After introduction of the case, this paper reviewed the clinical characteristics, diagnosis and treatment of Caroli's syndrome.［Chin J Contemp Pediatr, 2009, 11 (1):10-14］

OBJECTIVE: To understand the disease spectrum and treatment outcome on hospitalized neonates in China, a nationwide epidemiologic survey was performed. This paper reports the investigation results. METHODS: A retrospective study of 43 289 hospitalized neonates from 86 hospitals in 47 Chinese cities (22 provinces) between January 1, 2005 and December 31, 2005 was performed. RESULT: (1) The sex ratio of male to female was 1.73:1. (2) Premature infants accounted for 26.2% of the hospitalized neonates, which was higher than that reported in 2002 (19.7%). (3) The top three diseases that the neonates were susceptible to in turn were jaundice, pneumonia, and hypoxic-ischemic encephalopathy. (4) The percentage of pneumonia, meconium aspiration syndrome, and bilirubin encephalopathy in term infants was higher than that in premature infants, but the percentage of asphyxia, respiratory distress syndrome, and pulmonary hemorrhage in term infants was lower than that in premature infants. (5) The percentage of asphyxia, small for gestational age infant, and wet lung were higher in neonates whose mother had pregnancy induced hypertension. (6) The infants who recovered accounted for 63.9%, improved for 27.3%, requested own discharge for 7.6%, and died for 1.2%. Neonatal death of 46.4% occurred within 24 hrs after admission. CONCLUSIONS: The incidence of premature birth is increasing in hospitalized neonates. The neonatal deaths occur mostly within 24 hrs after admission.［Chin J Contemp Pediatr, 2009, 11 (1):15-20］

OBJECTIVE: A deficient interferon-gamma (IFN-gamma) response has been involved in the pathogenesis of severe respiratory syncytial virus (RSV) infection. Gene polymorphisms in IFN-gamma/A+874T have been associated with the susceptibility to asthma and might be related to disease severity of RSV infection. This study investigated the single nucleotide polymorphisms (SNPs) of IFN-gamma/A+874T in Han children in Wenzhou area and to explore the correlation between gene polymorphisms of IFN-gamma/A +874T and the susceptibility and disease severity of RSV bronchiolitis, as well as the effect of SNPs upon nasopharyngeal secretions (NPS) IFN-gamma and total serum IgE levels. METHODS: One hundred and fourteen hospitalized children with RSV bronchiolitis and 90 healthy controls were recruited. Sequence analysis was used for detecting the SNPs of IFN-gamma/A+874T. NPS IFN-gamma levels were measured using ELISA. Total serum IgE levels were assayed using the chemiluminescence method. RESULTS: IFN-gamma/A+874T gene polymorphisms were present in both the patient and the control groups. AA and AT genotypes were found in both groups, with a AA frequency of 82.5% vs 77.8% and a AT frequency of 17.5% vs 21.1% (P>0.05). The frequency of allele was 90.4% (A) and 9.6% (T) in the patient group, and 88.3% (A) and 11.7% (T) in the control group, respectively. There were no significant differences in the allele frequency between the two groups. Moreover, no difference was found both in NPS IFN-gamma and total serum IgE levels between AA and AT genotypes in the patient group. There were no significant differences in the variation of IFN-gamma/+874 between mild and moderate to severe cases. CONCLUSIONS: IFN-gamma/A+874T gene polymorphisms were present in Han children in Wenzhou area. Gene variations were not associated with the susceptibility and disease severity of RSV bronchiolitis as well as IFN-gamma and total serum IgE levels.［Chin J Contemp Pediatr, 2009, 11 (1):21-24］

OBJECTIVE: The frequency of the 22q11.2 deletion syndrome is increasing worldwide. The cardiovascular anomalies are one of the most frequent clinical manifestations in this syndrome. This study was designed to determine the frequency of 22q11.2 deletions in a prospectively ascertained sample from children with isolated conotruncal defects in China. METHODS: Twenty-four children with isolated conotruncal defects were prospectively enrolled and screened for the presence of 22q11.2 deletions using fluoresence in situ hybridization. The 24 patients consisted of two cases of persistent truncus arteriosus (PTA), five cases of pulmonary atresia/ventricular septal defect (PA/VSD), thirteen cases of tetralogy of Fallot (TOF), and four cases of double outlet right ventricle (DORV). RESULTS: Only 1 of the 24 patients had 22 q11.2 deletions. The frequency of 22q11.2 deletions (4.2%) was lower than that reported by other authors. CONCLUSIONS: Although 22q11.2 deletion is common in syndromic conotruncal anomalies, it is rare in isolated conotruncal anomalies.［Chin J Contemp Pediatr, 2009, 11 (1):25-28］

OBJECTIVE: To study the relationship of GYPC and TRIP3 gene expression and the prognosis of acute lymphoblastic leukemia (ALL) in children in order to explore the molecular biological mechanisms of recurrence and remission of ALL. METHODS: Thirty-eight newly diagnosed ALL children were enrolled. Of the 38 patients, 31 achieved complete remission (CR) and 12 relapsed. Semi-quantitative RT-PCR was employed to measure blood GYPC and TRIP3 gene expression. Twenty blood samples from normal children were used as controls. RESULTS: Blood GYPC expression in newly diagnosed ALL children was significantly higher than that in the control group (P<0.01) and the CR group (P<0.01). The expression of GYPC gene in the CR group was similar to that in the control group. Other than the control group (P<0.01) and the CR group (P<0.01), the GYPC expression of the relapse group was significantly higher than that in the newly diagnosed group (P<0.01). The CR group showed lower GYPC gene expression than the non-remission group before treatment (P<0.05). Blood expression of TRIP3 gene in the newly diagnosed and the relapse groups was significantly lower than that in the control group (P<0.05). The CR group had increased TRIP3 gene expression compared with the control group (P<0.01) as well as the newly diagnosed and the relapse groups (P<0.01). Of the 38 newly diagnosed ALL children, the patients with positive TRIP3 expression showed higher remission rate than those with negative TRIP3 (P<0.05). The TRIP3 gene expression before treatment in patients who achieved CR was higher than that in non-remission patients (P<0.05). CONCLUSIONS: A high GYPC gene expression is associated with an unfavorable outcome, in contrast, a high TRIP3 gene expression is associated with a favorable outcome in childhood ALL.［Chin J Contemp Pediatr, 2009, 11 (1):29-32］

OBJECTIVE: To evaluate the therapeutic effects of a combined immunotherapy, high-dose immunoglobulin (HDIG) plus cyclosporine A (CsA) plus prednisone (P), in children with aplastic anemia (AA) and to explore the association of peripheral blood lymphocyte subsets, peripheral blood cells and marrow CD34+ cells with therapeutic effects in AA. METHODS: The clinical data of 46 children with AA and who received the combined immunotherapy of HDIG + CsA + P were retrospectively studied. RESULTS: Of the 46 children with AA, 31 (67.4%) were responded to the combined immunotherapy. The binary logistic regression analysis showed low absolute neutrophil count (B=4.703, P<0.05), low percentage of peripheral blood CD4+ cells (B=0.142, P<0.05) and low ratio of peripheral blood CD4+/CD8+ （B=2.945, P<0.05）were associated with poor therapeutic effects. The ratio of CD34+/karyocytes of bone marrow in children with AA was lower than that in normal individuals, but it was not significantly related to the therapeutic effect. CONCXLUSIONS: The combined immunotherapy (HDIG+ CsA+P) was effective in children with AA. The absolute neutrophil count, the percentage of peripheral blood CD4+ and the ratio of peripheral blood CD4+/CD8+ were important prognostic factors in AA.［Chin J Contemp Pediatr, 2009, 11 (1):33-36］

OBJECTIVE: To study the roles of serum and urinary interleukins (IL)-13Rα2, IL-4, IL-6, IL8 and tumor necrosis factor-α(TNF-α) in pediatric Henoch-Schonlein purpura (HSP). METHODS: Serum and urinary levels of IL-13Rα2, IL-4, IL-6, IL-8 and TNF-α were examined using ELISA in 52 children with HSP and 45 healthy children. The results were compared between the two groups. RESULTS: Serum levels of IL-13Rα2, IL-4, IL-6, IL-8 and TNF-α in HSP patients with or without renal lesions were higher than those in the control group (P< 0.01 or 0.05). Urinary levels of IL-6 and TNF-α in HSP patients without renal lesions were higher than those in the control group (P<0.05). Except for urinary levels of IL-6 and TNF-α, urinary IL-13Rα2 levels in HSP patients with renal lesions (HSPN) were higher than those in the control group (P<0.05 ). CONCLUSIONS: Cytokines IL-13Rα2, IL-4, IL-6, IL-8 and TNF-α may play roles in the pathogenesis of pediatric HSP/HSPN.［Chin J Contemp Pediatr, 2009, 11 (1):37-40］

OBJECTIVE: Multilocular brain abscess in children is a serious neurosurgical emergency and remains a serious, life-threatening disease. This study evaluated the role of neuroendoscopy in treating multilocular brain abscess in children. METHODS: Between January 2002 and June 2007, 16 children with multilocular brain abscess underwent an operation using a pure endoscopic procedure. RESULTS: Increased intracranial pressure was relieved after operation in the 16 patients. CT/MRI after operation showed the abscess cavities disappeared and only the residual abscess walls existed in the 16 patients. Fourteen patients were followed up for 6 months to 5 years after surgery. Abscess walls disappeared in 13 patients and abscess recurred only in 1 patient. CONCLUSIONS: Neuroendoscopy for treatment of multilocular brain abscess is safe and effective in children.［Chin J Contemp Pediatr, 2009, 11 (1):41-43］

OBJECTIVE: To study the changes and significance of NF-kappa B activation in peripheral blood mononuclear cells (PBMC) of children with epilepsy. METHODS: NF-kappa B activation in PBMC was assayed by the flow cytometry in 32 healthy children and 64 children with epilepsy before and after treatment. The 64 epileptic children were subdivided into three groups: systemic seizure, partial seizure and unknown classification. RESULTS: NF-kappa B activation in PBMC in three epilepsy subgroups were significantly higher than that in healthy controls. The systemic seizure group showed significantly increased NF-kappa B activation in PBMC compared with the partial seizure group (P<0.01) and the unknown classification group (P<0.05). After treatment NF-kappa B activation in PBMC in three epilepsy subgroups was significantly reduced (P<0.01). CONCLUSIONS: NF-kappa B activation in PBMC increased in children with epilepsy, and it was positively correlated with the severity of seizures.［Chin J Contemp Pediatr, 2009, 11 (1):44-46］

OBJECTIVE: To examine plasma adiponectin (ADPN) and tumor necrosis factor-alpha (TNF-α) levels and their correlation in children with obesity in order to investigate the roles of both in the development of childhood obesity. METHODS: One hundred and forty-seven children with obesity and 118 normal children who were randomly sampled from five primary schools from the Kaifu District in Changsha were enrolled. Physical shape indexes, including height, weight, waist circumference, hip circumference, and waist to hip ratio (WHR) were measured. Body mass index (BMI) was calculated. Blood pressure was measured. Percentage of body fat (%BF) was measured with dual energy X-ray absorptiometry. Plasmal levels of ADPN and TNF-α were detected using ABC-ELISA. Blood concentrations of triglycerides (TG), total cholesterol (TC), high density lipoprotein-cholesterol (HDL-C) and low density lipoprotein-cholesterol (LDL-C) were measured by automatic biochemistry analyzer. Fasting blood glucose level was measured by glucose oxidase method. Fasting blood insulin level was assayed by radioimmunity. Homeostasis model assessment for insulin resistance (HOMA-IR) was performed. RESULTS: Plasma ADPN levels in obese children significantly decreased compared with those in normal children (8.12±2.54 mg/L vs 12.22±4.68 mg/L; P<0.05), and had a negative correlation with plasma TNF-α levels, BMI, WHR and HOMA-IR (P<0.01), and with %BF, fasting insulin, systolic blood pressure and TG (P<0.05). Plasma TNF-α levels in obese children significantly increased compared to normal children (171.38±34.33 ng/L vs 91.07±21.60 ng/L; P<0.01) and positively correlated with BMI, WHR, %BF, fasting insulin, HOMA-IR, TG and systolic blood pressure (P<0.01), and negatively with HDL (P<0.05). Multiple stepwise regression analysis showed that ADPN, BMI and TNF-α were main influential factors for %BF (R2=0.926, P<0.01). There was a significant interaction between ADPN and TNF-α (P<0.05). CONCLUSIONS: Plasma ADPN levels decreased and plasma TNF-α levels increased in children with obesity and both were main influential factors for %BF in children. There was an interaction between ADPN and TNF-α, suggesting that they both participate in the development of childhood obesity. ［Chin J Contemp Pediatr, 2009, 11 (1):47-50］

OBJECTIVE: Alveolar epithelium impairment is one of pathological changes associated with chronic lung disease (CLD). Hoxb5 is one of the few homeobox genes strongly expressed in the developing lung. This study investigated the expression of HoxB5, SPC and AQP5 in rats with CLD in order to explore the role of Hoxb-5 in impairment and reparation of alveolar epithelium. METHODS: Eighty neonatal rats were randomly exposed to hyperoxia (model group ) or to room air (control group) (n=40 each). The CLD model was induced by hyperoxia exposure. The expression of HoxB5, SPC and AQP5 protein and mRNA in the lung tissue was detected by immunohistochemistry and RT-PCR 1, 3, 7, 14 and 21 days after exposure. RESULTS: In the model group HoxB5 expression significantly decreased 7, 14 and 21 days after hyperoxia exposure. SPC expression decreased 3 days after hyperoxia exposure but increased significantly 7, 14 and 21 days after hyperoxia exposure as compared to the control group. AQP5 expression was progressively reduced with prolonged hyperoxia exposure. CONCLUSONS: Hyperoxia exposure may lead to alveolar epithelial cell (AEC) damage in neonatal rats. The increased SPC expression and decreased AQP5 expression suggested that the ability of differentiation and transformation of AECII into AECI decreased in neonatal rats with CLD. The decreased HoxB5 expression following hyperoxia exposure might contribute to a decreased ability of differentiation of AECII.［Chin J Contemp Pediatr, 2009, 11 (1):51-55］

OBJECTIVE: It has been proposed that nephrotic syndrome is a consequence of an imbalance between oxidant and anti-oxidant activity. Nephrin plays an important role in maintaining glomerular filtration barrier. This study aimed to explore the relationship between the expression of glomerular nephrin and oxidative stress reaction in rats with adriamycin (ADR)-induced nephrosis, and the protection of prednisone and vitamin E against renal injuries. METHODS: Nephrosis was induced by single intravenous injection of ADR (5 mg/kg). The prednisone intervention group was administered with prednisone (10 mg/kg daily) between 1 and 4 weeks after ADR injection. The vitamin E intervention group received vitamin E of 20 mg/kg daily from 1 week before ADR injection till to 4 weeks after ADR injection. Control rats were intravenously injected with normal saline. After 7, 14, 21 and 28 days of ADR injection, the indexes of oxidative stress reaction of the renal cortex, malondialdehyde (MDA), superoxide dismutase (SOD) and total antioxidative capacity (T-AOC), were measured using the chemical chromatometry. The protein expression of glomerular nephrin was measured by immunohistochemistry. RESULTS: Prednisone or vitamin E treatment reduced urinary protein from 14 days to 28 days after ADR injection. MDA levels of renal cortex increased, while renal activities of SOD and T-AOC as well as nephrin protein contents decreased in untreated nephrosis group from 7 days after ADR injection compared with those in the control rats. Compared with the untreated nephrosis group, prednisone treatment resulted in an increase in nephrin protein contents 28 days after ADR injection; Vitamin E treatment decreased renal MDA levels and increased renal activities of SOD and T-AOC and nephrin protein contents 28 days after ADR injection. Nephrin staining showed a sable linear-like pattern along the capillary loops of glomerulus in the control rats. Nephrin staining presented a light tan discontinuous short linear-like or punctiform pattern along the capillary loops of glomerulus in the untreated ADR group. Prednisone or vitamin E treatment ameliorated abnormal expression of nephrin induced by nephrosis. Glomerular nephrin expression level was negatively correlated with renal MDA level and positively correlated with renal activities of SOD and T-AOC. CONCLUSIONS: A reduction of glomerular nephrin expression is closely related to oxidative stress reaction. Prednisone and vitamin E have protective effects against renal injuries induced by ADR in rats.［Chin J Contemp Pediatr, 2009, 11 (1):56-60］

OBJECTIVE: To study the effect of H2O2 on the proliferation and apoptosis of endothelial progenitor cells (EPCs) and the antogonistic effects of catechin on the cell apoptosis induced by H2O2 in rats. METHODS: Immuno-fluoreascence assay was applied to detect CD34, CD133 and VEGFR-2 expression. EPCs of generation 2 were divided into control cells, H2O2-treated cells and catechin-H2O2-treated cells (H2O2: 100 mg/L; catechin: 10 mg/L). Genomic DNA was extracted by the conventional method after intervention for the analysis of apoptosis ladder pattern. The MTT assay was applied to detect proliferation rate of EPCs. RESULTS: The cultured cells at day 10 expressed CD34, CD133 and VEGFR-2. DNA apoptosis ladder pattern appeared in H2O2 -treated cells 2 days after intervention. After 3 days of intervention DNA apoptosis ladder pattern appeared in both H2O2-treated cells and H2O2-catechin-treated cells, with more ladders and grayer scale in H2O2 -treated cells. Compared with the controls, H2O2-treated cells and H2O2-catechin-treated cells showed significantly decreased proliferation rate (P<0.01), with the lowest proliferation rate at the 2nd day (P<0.05). The H2O2-catechin-treated cells showed increased proliferation rate than H2O2-treated cells at the 1st, 2nd and 3rd days. CONCLUSIONS: H2O2 may impair EPCs proliferation and induce EPCs apoptosis. Catechin may increase the capacity of EPCs for the resistance to apoptosis induced by H2O2.［Chin J Contemp Pediatr, 2009, 11 (1):61-64］

OBJECTIVE: Immunosuppressant tacrolimus (FK506) has shown neuroprotective effects on hypoxic-ischemic brain damage (HIBD) in the adult animal model. This study investigated whether FK506 has a protection against HIBD in neonatal rats by examining growth-associated protein-43 (GAP-43) expression in the hippocampus. METHODS: Ninety-six seven-day-old Sprague-Dawley rats were randomly divided into three groups: sham-operation, HIBD and FK506 intervention group. HIBD was induced in the later two groups. The FK506 intervention group was intraperitoneally injected with FK506 immediately after HIBD, at a dosage of 1 mg/kg daily, for three days. The HIBD group was injected with normal saline. Immunohistochemical technical was applied to examine GAP-43 expression in the hippocampus 24 and 72 hrs and 7 and 14 days after HIBD. RESULTS: Compared with the HIBD group, hematoxylin-eosin staining showed attenuated neuronal necrosis in the FK506 intervention group. In the HIBD group, the expression of GAP-43 increased significantly 72 hrs, and 7 and 14 days after HIBD compared with that in the sham-operation group. The GAP-43 expression in the FK506 intervention group was significantly higher than that in the HIBD group 72 hrs and 7 days after HIBD. CONCLUSIONS: FK506 might have neuroprotective effects against HIBD in neonatal rats.［Chin J Contemp Pediatr, 2009, 11 (1):65-68］

OBJECTIVE: To investigate the effects of ghrelin on the proliferation and differentiation of 3T3-L1 preadipocyte, and study the possible mechanisms. METHODS: 3T3-L1 preadipocytes were cultured in vitro. The proliferation potentials of 3T3-L1 preadipocytes that were treated with different concentrations of ghrelin were evaluated by MTT methods. The levels of c-myc and thymidine kinase mRNA were detected using RT-PCR. 3T3-L1 preadipocytes were differentiated into the matured adipocytes with insulin (INS) or ghrelin. The morphological changes of 3T3-L1 adipocytes were observed and the differentiation rate was assayed by oil-red O staining. Total RNA was extracted from adipocytes at various times, and the levels of peroxisome proliferation activated receptor γ (PPARγ) and CAAT/enhancer binding protein(C/EBPα) mRNA expressions were detected using RT-PCR. RESULTS: Ghrelin at concentrations of 10-7 to 10-15 mol/L significantly stimulated preadipocyte proliferation (P<0.05). The levels of c-myc and thymidine kinase mRNA significantly increased in 3T3-L1 preadipocytes with 10-9 mol/L and 10-11 mol/L ghrelin treatment (P<0.01). The 3T3-L1 preadipocytes treated with 10-11 mol/L ghrelin had lots of lipid droplets in the cytoplasma, but the differentiation rate was lower than those treated with INS. Ghrelin of 10-11 mol/L significantly increased the mRNA expression of PPARγ and C/EBPα in the course of 3T3-L1 preadipocyte differentiation, compared with the normal control group (P<0.05). The PPARγ and C/EBPα mRNA expression increased with the prolonged differentiation of preadipocytes induced by ghrelin or INS. There were significant differences in the levels of PPARγ and C/EBPα mRNA expression between the 2nd and 8th days of differentiation（P<0.01）. CONCLUSIONS: Ghrelin promotes the proliferation and differentiation of 3T3-L1 preadipocytes. The proliferation of 3T3-L1 preadipocytes induced by ghrelin may be associated with increased c-myc levels. Ghrelin may promote differentiation of 3T3-L1 preadipocytes by increasing mRNA expression of PPARγ and C/EBPα, thus enhances the sensitivity of adipocytes to INS.［Chin J Contemp Pediatr, 2009, 11 (1):69-73］