OBJECTIVE: To investigate the antimicrobial resistance of Streptococcus pneumoniae (S.pneumoniae) isolated from Chinese children with pneumonia.METHODS: Hypopharyngeal aspirate specimens were collected from hospitalized children with pneumonia who were admitted to the children's hospital located in Beijing, Shanghai, Guangzhou or Shanghai from Feburary 16, 2006 to Feburary 16, 2007. The minimum inhibitory concentration (MIC) of S.pneumoniae isolates against penicillin, amoxicillin, cefuroxime (sodium), ceftriaxone, erythromycin, vancomycin, ofloxacin and imipenem was determined by E-test method. RESULTS: A total of 279 S.pneumoniae isolates were obtained. Eighty-six percent of the isolates were not susceptive to penicillin, and 23.3% was resistant to penicillin. The rate of susceptibility of the isolates to amoxicillin was 92.1%, and to cefuroxime and ceftriaxone was 19.0% and 75.3%, respectively. The isolates also showed a high susceptibility to vancomycin (99.6%) and ofloxacin (97.8%). Seventeen point six percent of the isolates were not susceptive to imipenem, and most of those were intermediate. Almost of all isolates were resistant to erythromycin. There were some distinct regional differences in the susceptibility to antimicrobials tested except for erythromycin, vancomycin and ofloxacin. CONCLUSIONS: The S.pneumoniae isolates from Chinese children with pneumonia were susceptive to amoxicillin, vancomycin and ofloxacin, but were not susceptive or resistant to penicillin, cefuroxime and erythromycin. The isolates kept susceptibility to ceftriaxone and imipenem to a certain extent.

OBJECTIVE: Bacterial meningitis is a kind of central nervous system infection with a high incidence, disability and fatality in children. Prompt diagnosis and treatment are associated with an improved prognosis. Low positive rate of bacterial cultures of the cerebrospinal fluid (CSF) makes it difficult to make a definite diagnosis. This experiment aimed to investigate a proteome profile of normal CSF of Chinese children by two-dimensional polyacrydamide gel electrophoresis (2-DE), and to sieve the disease-specific proteins of Staphylococcus epidermidis meningitis (SeM) to provide basis for early diagnosis and treatment of SeM.METHODS: Four mL CSF samples were obtained respectively from SeM and normal children. The separated proteins with immobile pH gradient (IPG) 2-DE technology and protein spots were visualized by Coomassie Brilliant Blue staining. The stained 2-DE gels were scanned on the Imagescanner and pictures were obtained through Labscan software. The images were analyzed with PDQuest software and the differences of protein spots were compared between the SeM and normal children. RESULTS: Mean protein spots of the 2-DE gels were 438 and 425 in the SeM and normal groups respectively. Twenty-five protein spots only occurred in normal CSF and 12 spots only occurred in the SeM group. The expression of 6 protein spots showed up-regulation and that of 19 showed down-regulation in the SeM group compared with that in the normal group. CONCLUSIONS: A 2-DE profile of CSF proteome was successfully established in SeM and normal children through proteomic technique. By the differentiated analysis of these CSF 2-DE gels, the differences of CSF proteome profiles were found between SeM and normal children. Future analysis and identification of these spots will contribute to find out the disease specific proteins of SeM and to provide basis for early diagnosis and therapy of this disorder.

OBJECTIVE: Vascular endothelial cell injury contributes to the pathogenesis of viral encephalitis. This study was designed to investigate the roles of vascular endothelial growth factor (VEGF) and vascular cell adhesion molecule-1(VCAM-1) in cerebral spinal fluid (CSF) in the pathogenesis of viral encephalitis and in the evaluation of the severity and the prognosis of viral encephalitis in children. METHODS: CSF VEGF and VCAM-1 levels were measured using ELISA in 65 children with viral encephalitis and 20 age-matched controls (10 cases of epilepsy and 10 cases of congenital abnormality). RESULTS: CSF levels of VEGF and VCAM-1 in the viral encephalitis group in the acute phase were significantly elevated compared with those in the congenital abnormality (P<0.01) and the epilepsy groups (P<0.05). CSF levels of VEGF and VCAM-1 in the viral encephalitis group in the recovery phase decreased significantly and were similar to the levels of the epilepsy group, but remained higher than those in the congenital abnormality group (P<0.05). There was a positive correlation between CSF levels of VEGF and VCAM-1 in the viral encephalitis group in the acute and recovery phases. CSF levels of VEGF and VCAM-1 were positively correlated to CSF protein contents and the degree of MRI abnormality in the viral encephalitis group. CONCLUSIONS: VEGF and VCAM-1 may participate in the pathogenesis of viral encephalitis. Detection of the two parameters may be helpful to the evaluation of the severity and prognosis of viral encephalitis.

OBJECTIVE: To study the prognostic factors for events-free survival (EFS) in children with acute non-mature B-lymphoblastic leukemia.METHODS: One hundred and sixty-one children with newly diagnosed acute non-mature B-lymphoblastic leukemia received the ALL-XH-99 protocol treatment. Their medical data, including clinical, biological and molecule features, early responses to treatment (bone marrow evaluation on the 19th day of induction therapy), minimal residual disease (MRD) in bone marrow after remission induction therapy, the risk grade of disease before the beginning of chemotherapy and the outcome, were retrospectively studied.RESULTS: Univariable analysis indicated that the gender and P170 levels before therapy had no effect on the outcome. Age, initial white blood cell count (WBC), prednisone response, early response to treatment, fusion genes (BCR/ABL or MLL/AF4) and MRD level were significantly related to the EFS (P<0.01). Immunophenotype, myeloid-associated antigen and the risk grade of disease were also related to the EFS (P<0.05). Multivariable analysis showed that WBC ≥50×109/L, Cμ positive, BCR/ABL or MLL/AF4 positive and MRD positive (≥0.01%) were risk factors for the poor prognosis (P<0.01). The early response to treatment was important to modify the therapy protocol. CONCLUSIONS: WBC ≥50×109/L, Cμ positive, BCR/ABL or MLL/AF4 positive and MRD positive have important prognostic values in childhood acute non-mature B-lymphoblastic leukemia. Early response to treatment is an important index for modifying the chemotherapy protocol.

OBJECTIVE: To investigate the characteristics of gene expression of surfactant protein A in Chinese premature infants and the association between surfactant protein A and the risk of neonatal respiratory distress syndrome (RDS).METHODS: Vein-blood samples (2 mL) from 18 Chinese premature infants with RDS and 28 controls were assayed for SP-A genotypes 6A2, 6A3, 1A0 and 1A1 by SSCP.RESULTS: The frequency of allele distribution of SP-A1 allele 6A2 and 6A3 was 0.50 and 0.056 respectively in the RDS group and was 0.214 and 0.107 in the control group. Compared with the controls, SP-A1 allele 6A2 was over-represented in the RDS group (P<0.05). In contrast, SP-A1 allele 6A3 tended to be under-represented in the RDS group but there was no statistical difference when compared with the controls. The frequency of allele distribution of SP-A2 allele 1A0 and 1A1 was 0.722 and 0.667 respectively in the RDS group and was 0.679 and 0.821 respectively in the control group. There were no significant differences in the distribution frequency of SP-A2 allele 1A0 and 1A1 between the two groups. In the infants born at gestation >32 weeks, SP-A1 allele 6A2 was over-represented in the RDS group compared with the control group (frequency: 0.56 vs 0.15; P<0.05). CONCLUSIONS: The frequency of SP-A1 allele 6A2 and 6A3 was low, in contrast, the frequency of SP-A2 allele 1A0 and 1A1 was high in normal Chinese premature infants. SP-A1 allele 6A2 may be a susceptible gene for RDS.

OBJECTIVE: To explore the value of electroencephalogram (EEG) in early diagnosis of brain injury in neonates with asphyxia.METHODS: EEG examination was performed in 49 neonates with asphyxia (mild: n=9; severe: n=40) within 6 hrs of their births. Of the 49 asphyxiated neonates, 33 had concurrent HIE, including 20 cases of mild, 9 cases of moderate and 4 cases of severe HIE. RESULTS: Twenty-one (63.6%) out of the 33 patients with HIE showed abnormal EEG, but only one (6.3%) in the asphyxia group without HIE. All of 13 patients with moderate-severe HIE showed abnormal EEG. The degree of EEG abnormality in neonates with HIE was consistent with the clinical grading of HIE. The neonates whose EEG showed electrical silence and burst suppression and the abnormalities were kept unrecoverable for more than 2 weeks had very poor prognosis.CONCLUSIONS: EEG can reflect brain injury caused by neonatal asphyxia and the severity of brain injury. It may be useful for early diagnosis of brain injury following asphyxia in neonates.

Hypophosphatasia is a rare inborn disease of metabolism. This paper reviewed its pathogenesis, forms, clinical manifestations, differential diagnosis, treatment and prognosis. Here a case of neonatal hypophosphatasia is reported. This baby was female (30 minutes old). Prenatal ultrasound showed disproportionate biparietal diameter and long bones of limbs in the baby. After birth, she presented with obvious craniomalacia, respiratory distress and cyanosis. Serum alkaline phosphatase level was significantly reduced. Both X-ray and autopsy showed extremely insufficient skeletal mineralization. Four days later she died of respiratory failure.

OBJECTIVE: To assess the values of serum fatty acid-binding protein （FABP) and brain natriuretic peptide (BNP） in pneumonia complicated by acute congestive heart failure (CHF) in children.METHODS: Serum levels of FABP and BNP were determined using ELISA in 36 children with pneumonia complicated by CHF (pneumonia group) and 28 healthy children (control group).RESULTS: Serum levels of FABP and BNP in the pneumonia group at the acute stage were significantly higher than those in the control group and those at the recovery stage (P<0.01). Compared with the control group, serum levels of FABP and BNP in the pneumonia group at the recovery stage increased significantly (P<0.01). At the acute stage, 35 patients (97.2%) showed increased serum FABP level but 28 (77.8%) showed increased serum BNP level (P<0.05) in the pneumonia group. At the recovery stage, the incidence of abnormal serum FABP (72.2%) was significantly higher than that of BNP (44.4%) in the pneumonia group (P<0.05).CONCLUSIONS: Serum levels of FABP and BNP can be regarded as biochemical markers of myocardial damage in children with pneumonia complicated by CHF and serum FABP appears to be a more sensitive one. Serum FABP and BNP remained at higher levels through the recovery stage, suggesting that myocardial damage existed though the clinical symptoms were improved at the stage.

OBJECTIVE: Neonatal respiratory failure (RF) is a serious clinical problem associated with high mortality. This study aimed to investigate the incidence and outcome of neonatal RF in the Children's Hospital of Hebei Province. METHODS: The medical data of 304 RF infants who were admitted to the neonatal intensive care unit (NICU) of the Children's Hospital of Hebei Province between March 2004 and February 2005 were prospectively studied. RESULTS: The incidence of RF was 35.7% in the NICU during the 12-month period. Respiratory distress syndrome (28.4%), amniotic fluid aspiration syndrome (22.9%), and pneumonia (14.7%) were leading diseases resulting in RF. Of the 304 pati-ents, 17 (5.6%) died of RF, 96 (31.6%) discontinued therapy on account of various causes and 191 (62.8%) recovered or improved. The mean length of hospital stay was 14.9±7.1 days and the mean medical cost was 7 977±3 426 CNY for the 191 survivors.CONCLUSIONS: The morbidity, mortality and medical costs of neonatal RF are high in Hebei Province. It is essential to improve the quality in respiratory therapy and perinatal care in order to reduce the morbidity and mortality related to high-risk pregnancies.

OBJECTIVE: This research reported the experience of early surgical treatment for infants with large atrial septal defects (ASD) or ventricular septal defects (VSD) complicated by pneumonia.METHODS: Between January 2003 and January 2008, 39 infants with large ASD or VSD complicated by pneumonia were admitted to the Second Xiangya Hospital. Thirty-six patients underwent surgical repair within 7-10 days after pneumonia had been controlled. Mean age was 5.4±3.4 months and mean weight was 4.7±1.6 kg in the 36 patients. Three patients received conservative treatment due to uncontrolled lung infections.RESULTS: Of the 36 patients, 33 had successful surgery and 3 (8.3%) died of serious low cardiac output (n=1) or respiratory failure due to congenital tracheostenosis (n=2). The 33 survivors showed normal growth and development in a 6 month-5 year follow-up. Of the 3 patients receiving conservative treatment, 1 died of cardiopulmonary failure and 2 were discharged after the symptoms had been improved. CONCLUSIONS: With increasing medical experience and technique, early surgical operation may be performed with good outcomes in infants with large ASD or VSD complicated by pneumonia.

OBJECTIVE: To study the value of bronchofibroscopy in the etiologic identification of chronic cough in children.METHODS: Under local anesthesia of lidocaine, bronchofibroscopy was performed in 118 children with chronic cough of unknown origin (73 males and 45 females). Their ages ranged from 3 months to 13 years. RESULTS: The cause of chronic cough was identified in 115 cases. The most common cause was respiratory infection (n=39)，followed by bronchial foreign bodies （n=19）, upper airway cough syndrome (n=17), bronchial asthma or cough variant asthma (n=17）, bronchomalacia (n=7), bronchial congenital malformation (n=5), primary ciliary dyskinesia (n=3), gastro-esophageal reflux (n=3), bronchial tumor (n=2), bronchial tuberculosis (n=1), pulmonary fibrosis (n=1) and idiopathic pulmonary hemosiderosis (n=1). CONCLUSIONS: Bronchofibroscopy is useful in the etiologic identification of chronic cough in children.

OBJECTIVE: To study the features of interictal epileptiform discharges (IED) during sleep and wakefulness in children with epilepsy.METHODS: The polysomnography, active EEG and video EEG were performed on 48 children with epilepsy during the whole night, and wakefulness of pre- and post-sleep. The epileptiform sharp/spike discharge indexes during sleep and wakefulness were recorded. The positive rate of IED in focal and generalized epilepsy was compared.RESULTS: Of the 48 patients, 25 showed IED, including 9 cases (36.0%) in the generalized seizure group and 16 cases（64.0%） in the focal seizure group（P<0.05）. The epileptiform sharp/spike discharge indexes in the whole non-rapid eye movement (NREM) sleep stage (stages S1-S4: 21.13±19.96, 19.59±17.76, 22.85±18.99, and 20.37±16.63) were significantly higher than that in the wakefulness stage (8.20±6.21) (P<0.05). The discharge index in the S3 stage during NREM sleep was higher than that during the rapid eye movement (REM) sleep (22.85±18.99 vs 12.91±10.95; P<0.05).CONCLUSIONS: The positive rate of IED in the focal seizure group was higher than that in the generalized seizure group. Sleep, especially NREM sleep, facilitates IED in children with epilepsy.

OBJECTIVE: Age, body weight and dose have been shown as important influencing factors for sodium valproate plasma concentrations. However it is unclear whether there is interaction among them and whether the interaction could influence sodium valproate plasma concentrations. This study aimed to evaluate the influence of age, body weight and dose on plasma concentrations of sodium valproate and the interaction among them.METHODS: One hundred and thirty-two children with epilepsy (age: 4 months-6 years, weight: 5-25 kg) were enrolled. Sodium valproate was administered at the dosage of 10-30 mg/kg.d. Plasma concentrations of sodium valproate were measured by high-performance liquid chromatography 3-5 days after administration. The relationship of sodium valproate plasma concentrations with age, body weight, and dose of sodium valproate was examined using variance analysis, pearson correlation analysis and stepwise regression analysis. RESULTS: Age (F=8.630, P<0.01), body weight (F=3.650, P<0.05) and dose of sodium valproate (F=11.720, P<0.01) were influencing factors for sodium valproate plasma concentrations. The interaction between age and oral dose (F=2.484, P<0.05) and the interaction of age and body weight with oral dose (F=4.923, P<0.01) had significant effects on sodium valproate plasma concentrations. Stepwise regression analysis showed that dose of sodium valproate and body weight were entered to the regression equation.CONCLUSIONS: Age, body weight and dose of sodium valproate as well as the interactions between age and dose and between age, body weight and dose were influencing factors for valproate plasma concentrations.

OBJECTIVE: To evaluate the effectiveness of AML-XH-99-M3 protocol for treatment of acute promyelocytic leukemia (APL) in children.METHODS: Thirty-three children with APL received AML-XH-99-M3 protocol treatment. The event-free survival (EFS), disease-free survival (DFS), and overall survival (OS) were evaluated by the Kaplan-Meier medthod with SPSS13.0 software.RESULTS: Thirty patients (90.9%) achieved a complete remission (CR) after one course of treatment. The total CR rate was 100%. Six patients (18.2%) relapsed in an average of 29.17 months (16-38 months). Two patients (6.1%) died. The 7-year EFS and DFS in the 30 patients were 73.4±9.4%. The overall survival rate was 91.2±6.0%. The difference of EFS was observed in patients receiving intermittent all-trans-retinoic acid (ATRA) administration or not in the maintenance therapy (88.9±10.5% vs 62.5±13.6%) (P<0.05).CONCLUSIONS: The AML-XH-99-M3 protocol for the treatment of APL produced a higher CR rate and higher EFS, DFS and OS rates in children. Intermittent administration of ATRA in the maintenance therapy can improve EFS rate.

OBJECTIVE: To study the role of minimal residual disease (MRD) in the evaluation of therapeutic effectiveness of childhood B-cell acute lymphoblastic leukemia (ALL)．METHODS: MRD testing was performed in 124 children with B-cell ALL, who were newly diagnosed and enrolled in the ALL-XH-99 treatment protocol from September 2001 to April 2005．MRD was determined by 4-color flow cytometry in the different time points during the treatment period．RESULTS: After induction therapy, 103, 13 and 8 patients showed MRD <0.01%, 0.01%-0.1% and >0.1%, respectively. The 5-year relapse-free survival (RFS) in the patients with MRD <0.01%, 0.01%-0.1% and >0.1% was 88.9±3.9%, 70.0±14.5% and 0%, respectively and the 5-year event-free survival (EFS) was 82.4±4.4%, 21.2±18.0% and 0%, respectively. There were significant differences in the RFS and EFS among the patients with different MRD levels (P<0.01). Within half a year after induction remission, the 5-year RFS in patients with MRD negative (<0.01%) and positive was 87.7±4.1% and 58.3±14.2%, respectively (P<0.01) and the 5-year RFS was 80.7±4.6% and 25.6±13.8%, respectively (P<0.01). After half a year with induction remission, the patients with MRD negative and positive also showed statistical differences in the 5-year RFS (92.0±3.6% vs 48.5±15.5%; P<0.01) and EFS (85.6±4.5% vs 21.4±11.0%; P<0.01). Multivariate analysis confirmed that the MRD level after induction chemotherapy together with the reaction to prednisone, the bone marrow features on the 19th day of induction, and the fusion gene with BCR-ABL or MLL-AF4 had prognostic significance in childhood B-cell ALL.CONCLUSIONS: The MRD level in the whole course of therapy is an important outcome indicator in childhood B cell ALL.

OBJECTIVE: Identifying the stressors, coping strategies, and psychosocial state of children with chronic illness would be very useful to help them to adapt to chronic medical conditions. This study aimed to investigate the stressors, coping strategies, and psychosocial state of Chinese children with chronic illness.METHODS: Two hundred and three children with chronic illness and aged 8-16 years were administered a semi-structured interview for the identification of stressors. Children's coping strategies and psychosocial state were investigated by the Coping with a Disease (CODI) scale, the Screen for Child Anxiety Related Emotional Disorders (SCARED) scale, and the Depression Self-rating Scale for Children (DSRSC). RESULTS: The stressors in children with chronic illness mainly included four aspects: school performance, medication and treatment, daily life, and peer relationships. "Wishful thinking" was the most common coping strategy, followed by "acceptance" . "Negative emotional reaction" was rarely seen in children with chronic illness. The scores of anxiety and depression scales of children with chronic illness were higher than those of the norm. The prevalence of anxiety disorders was 43.8%, the prevalence of depression disorders was 30.0%, and 26.1% of the children had both anxiety and depression disorders.CONCLUSIONS: Children with chronic illness have many stressors. Though they usually use active coping strategies, the prevalence of anxiety disorders and the prevalence of depression disorders were high.

OBJECTIVE: To study the changes and roles of plasma thromboxane A2 (TXA2) and prostaglandin I2 (PGT2) levels and their ratio in Henoch-Schonlein purpura nephritis (HSPN) in children. METHODS: Plasma levels of TXA2 and PGI2 were measured using ELISA in 45 children with HSPN and 20 healthy children. RESULTS: Plasma TXA2 level was significantly higher, while plasma PGI2 level was significantly lower in HSPN children in the acute phase than in the control (P<0.01). The ratio of TXA2/PGI2 in HSPN children in the acute phase was statistically higher than in the control (9.55±3.56 vs 0.87±0.21; P<0.01). In the convalescence phase, plasma TXA2 level remained higher and plasma PGI2 level was elevated and higher than in the control, so the ratio of TXA2/PGI2 was reduced to normal level.CONCLUSIONS: The imbalance of TXA2 and PGI2 may be involved in the development of renal damage in children with HSPN. The balance of TXA2 and PGI2 contributes to renal recovery.

OBJECTIVE: In some hospitals an adult colonoscope is used for colon examination in children because they do not have child colonscope equipment. This has some disadvantages and this paper reports the experience for colon examination in children with an adult gastroscope instead of an adult colonoscope .METHODS: One hundred and three children aged from 1.3 to 14 years who required routine colon examination were randomly assigned to adult gastroscope (n=49) and adult colonoscope groups (n=54).RESULTS: There were no significant differences in the success rate of implantation into the ileocecum between the gastroscope and colonoscope groups (93.9% vs 94.4%; P>0.05). The average time of implantation into the ileocecum in the gastroscope group was shorter than that of the colonoscope group (5.2±1.1 min vs 7.3±2.9 min; P<0.05). Seventeen patients showed implantation-related complications in the colonoscope group but only 5 patients in the gastroscope group (P<0.01).CONCLUSIONS: An adult gastroscope appears to be safer and more feasible than an adult colonoscope for colon examination in children.

OBJECTIVE: To investigate calcium, iron and magnesium intakes of preterm infants' mothers before and during pregnancy and calcium, iron and magnesium levels of preterm infants and their mothers in order to provide basis for studying the effect of nutritional factors on the occurrence of prematurity. METHODS: Two hundred and forty matched cases ( preterm infants and their mothers) and controls (term infants and their mothers) were recruited. A nutritional survey of calcium, iron and magnesium intakes was performed in the mothers before and during pregnancy. Calcium, iron and magnesium levels in maternal plasma and in cord blood, placenta, breast milk, meconium, and amniotic fluid were measured with axial view inductively coupled plasma optical emission spectrometry (ICP-OES).RESULTS: Iron and magnesium intakes in preterm infants' mothers were significantly less than those in term infants' mothers before pregnancy (P<0.05). Iron and calcium intakes in preterm infants' mothers were also significantly less than those in term infants' mothers during pregnancy (P<0.05). Multivariate analysis of variance showed that iron and calcium levels of preterm infants'mothers were significantly lower than those of term infants' mothers (P<0.05). The preterm infants showed significantly lower iron and magnesium levels than term infants (P<0.05). Plasma levels of calcium, iron and magnesium in infants were positively correlated to maternal plasma levels of calcium, iron and magnesium (r=0.517, 0.622, 0.518, respectively; P<0.05).CONCLUSIONS: The iron and calcium levels of preterm infants' mothers were lower than those of term infants' mothers, and the iron and magnesium levels of preterm infants were lower than those of term infants. The exact relationship between calcium, iron and magnesium levels and intakes before and during pregnancy needs to be explored further.

OBJECTIVE: The development of pulmonary vascular bed is strongly flow-dependent. Abnormal pulmonary blood flow leads to pulmonary pathological changes. This study aimed to observe the pathological changes of small pulmonary arteries and alveoli in complex congenital heart defect with diminished pulmonary blood flow but without aortopulmonary collateral artery (APCA) and patent ductus arteriosus (PDA) in infants and young children. METHODS: Autopsy pulmonary specimens obtained from 5 children who died of non-cardiovascular diseases were used as the control group (age: 4-18 months). Fifty-six children (age: 4-36 months) with complex congenital heart defect with diminished pulmonary blood flow but without APCA and PDA served as the study group, including 34 cases of tetralogy of Fallot, 7 cases of double outlet right ventricle with pulmonary stenosis, 9 cases of single ventricle with pulmonary stenosis, 4 cases of tricuspid atresia with pulmonary stenosis and 2 cases of complete atrioventricular septal defect with pulmonary stenosis. Pulmonary specimen sections were stained by hematoxylin-eosin and Weigert-Van Gieson. Percentage of media thickness (MT%), percentage of media section area (MS%), number of small arterial per square centimeter (APSC), mean alveolar number (MAN), mean linear intercept (MLI), proportion of parenchyma area in total area (PPA%) and alveolar to small arterial ratio per unit area (AAR) were measured by morphologic quantitative analysis.RESULTS: MT% (10.93±2.87% vs 15.08±2.51%), MS% (18.97±5.56% vs 25.04±3.87%) and APSC (202.43±67.45 vs 441.69±65.29) decreased significantly in the study group compared with the control group (P<0.01). The internal diameter of small pulmonary artery (80.26±21.57 μm vs 58.53±10.29 μm; P<0.05), AAR (46.59±14.43 vs 34.46±4.98; P<0.01) and MLI (144.98±44.87 μm vs 108.39±20.76 μm; P<0.05) increased significantly compared with the control group.CONCLUSIONS: The media of small pulmonary arteries becomes thinner, the lumen of small pulmonary arteries becomes larger, and the number of small arterial per square centimeter and the mean alveolar number are reduced in infants and young children with complex congenital heart defect with diminished pulmonary blood flow but without APCA and PDA.

OBJECTIVE: To study the sleep time and the prevalence of sleep disorders in children at ages of 2-12 years in Changsha City. METHODS: A total of 3 756 children at ages of 2-12 years were randomly sampled from five districts of Changsha City from June 2006 to April 2007. A questionnaire survey was performed on their parents.RESULTS: The average daily sleep time in the subjects was 10.60 hrs. The average daily sleep time among different age groups (1 year as a group separation) was different. It was 12.26, 11.57, 11.33, 11.26, 10.95, 10.64, 10.62, 10.45, 10.28, 9.83 and 9.61 hrs respectively in the 11 age groups of 2 to 12 years of age. The prevalence of sleep disorders in children at ages of 2-12 years was 40.9%, including frequent sleep snoring (8.2%), choke/gargling (1.5%), sleep apnea (0.8 %), sleep inquietude (7.6%), mouth breathing (4.9%), hyperhidrosis (22.6%), limbs spasm (3.2 %), sleep teeth grinding (9.5 %), sleep talking (5.5 %), sleep walking (0.9 %), nocturnal enuresis (2.5%), waking up by choke (1.9%), remaining wakefulness in the night due to too much daytime sleep time (1.5%), going to sleep too early (2.1%), night awakenings (1.6%), and screeching or crying during sleep (1.8%). The prevalence of different sleep disorders was different in children between boys and girls and among different age groups. CONCLUSIONS: The average sleep time in children at ages of 2-5 years is less than the reference value recommended by the domestic child health care textbook. There is a higher prevalence rate of sleep disorders in children at ages of 2-12 years in Changsha City than the reported data in other cities.

OBJECTIVE: To study the effects of androgen on the expression of phosphacan and NG2 proteoglycan (NG2) and neurite regeneration in neonatal rats with hypoxic-ischemic brain damage (HIBD) and the potential mechanism underlying the protective effect of androgen against HIBD.METHODS: One hundred and twenty neonatal Sprague-Dawley rats were randomly divided into three groups: sham-operated, HIBD and androgen treatment. HIBD was induced by the ligation of left common carotid artery and hypoxia exposure. The androgen treatment group rats were injected with testosterone propionate (25 mg/kg) immediately after HIBD. Phosphacan and NG2 expression in the cortex and the hippocampus was detected with the immunohistochemical method 24 and 72 hrs and 7 and 10 days after hypoxia-ischemia (HI). The ultrastructure and neurite regeneration of neurons in the cortex and the hippocampus were observed under a transmission electron microscope.RESULTS: The neurite regeneration was obvious in the sham-operated group, but seldom in the HIBD group. The androgen treatment group showed increased neurite regeneration compared with the HIBD group. There were fewer phosphacan and NG2 positive cells in the cortex and the hippocampus in the sham-operated group. Phosphacan and NG2 expression in the cortex and the hippocampus was observed at 24 hrs, increased at 72 hrs, and peaked at 7 days after HI in the HIBD group and remained at a higher expression 10 days after HI than in the sham-operated group. The levels of phosphacan and NG2 expression in the cortex and the hippocampus in the androgen treatment group were significantly reduced compared with those in the HIBD group 24 and 72 hrs and 7 and 10 days after HI (P<0.01). CONCLUSIONS: Phosphacan and NG2 may be important inhibitory factors for neurite regeneration following HIBD in neonatal rats. The neuroprotection of androgen against neonatal HIBD is produced possibly through an inhibition of phosphacan and NG2 expression.

OBJECTIVE: To evaluate the brain pathological changes following exdogenous neural stem cells (NSCs) intraventrilar transplantation in neonatal rats with periventricular leukomalacia (PVL), and to explore the feasibility of NSCs transplantation for the treatment of PVL in premature infants. METHODS: NSCs were prepared from E14 embryonic rat brain. Two-day-old neonatal rats were randomly divided into six groups: PVL, PVL+culture medium, PVL+NSCs, sham operation, sham operation+culture medium, and sham operation+NSCs (18-21 rats each group).Intraventricular transplantation of exdogenous NSCs was performed 72 hrs after PVL induction or sham operation. The cerebral pathological evaluation was undertaken by light microscopy 7, 14 and 21 days after transplantation. RESULTS: The pathological changes in the cerebral white matter were gradually improved with the prolonged time after transplantation. After 21 days of transplantation, 50% of the cerebral white matter showed mild pathological changes and 50% of that showed severe pathological changes, with neuronal pathological scores of 1.28±0.86, in the untreated PVL group. In the PVL+NSCs group, 30% of normal white matter, 40% of mild and 30% of severe pathological changes in the white matter were observed, with neuronal pathological scores of 0.32±0.16, 21 days after transplantation. There were very significant differences in both of pathological changes in the cerebral white matter and neuronal pathological scores between the PVL and PVL+NSCs groups (χ2=10.7, P<0.01; F=29.664, P<0.01).CONCLUSIONS: Intraventricular transplantation of exdogenous NSCs can apparently improve cerebral white matter damage. It is suggested that intraventricular transplantation of NSCs is of a great potential feasibility for the treatment of PVL in premature infants.

OBJECTIVE: To investigate the short-term effects of flurothyl-induced neonatal recurrent seizures on γ-aminobutyric acid A receptor (GABAAR) α1 and β2 subunit expression in the rat brain, and to study the relationship between the alterations of GABAAR subunits in the developing brain and seizure-induced brain injury.METHODS: Sixty-four 7-day-old Sprague-Dawley rats were randomly divided into two groups: control and seizure. Seizures were induced by inhalant flurothyl daily for six consecutive days. The expression of GABAAR α1 and β2 subunits protein in the cerebral cortex and the hippocampus were detected by Western blot and immunohistochemistry method 1 and 7 days after recurrent seizures.RESULTS: Compared to the control, the accumulated optical density (AOD) of GABAAR α1 subunit immunoreactivity (IR) in the parietal cortex, the CA3-CA4 regions and the dentate gyrus in seizure rats increased significantly 1 day after recurrent seizures (P<0.05). The AOD of GABAAR α1 subunit IR in the parietal cortex, the CA1-CA4 regions and the dentate gyrus in seizure rats increased significantly 7 days after recurrent seizures compared with the control (P<0.05). The expression of GABAAR α1 subunit in the hippicampus and the cerebral cortex increased significantly in seizure rats compared with that in control rats 1 and 7 days after recurrent seizures. After 7 days of recurrent seizures, the AOD of GABAAR β2 subunit IR in the CA1-CA2 regions increased significantly in the seizure group compared with that in the control group (P<0.05), but the AOD of GABAAR β2 subunit IR in the thalamus decreased significantly in the seizure group compared with that in the control group (P<0.05). The expression of GABAAR β2 subunit protein in the hippocampus increased significantly in the seizure group compared with that in the control group 7 days after recurrent seizures (P<0.05). CONCLUSIONS: Recurrent neonatal seizures may result in the short-term alterations of GABAAR α1 and β2 subunits expression in the cerebral cortex and the hippocampus in rats, suggesting the alterations of GABAAR subunit expression may be related to the developing brain injury following recurrent seizures.

OBJECTIVE: To study the expression of heme oxygenase-1 (HO-1) gene and protein and the effect of budesonide (BUD) on the HO-1 expression in lung tissues in rats with asthma.METHODS: Fifty male Sprague-Dawley rats were randomly divided into 5 groups: normal control, asthma model, dexamethasone (DXM)-, hemin (HO-1 challenger)-or BUD-treated asthma. The asthma model was prepared by ovalbumin sensitization and challenge. The rats were sacrificed 24 hrs after the last challenge. The blood COHb content, and the total cell count and the percentage of differential cells in bronchoalveolar lavage fluid (BALF) were measured. The expression of HO-1 protein and mRNA in lung tissues was detected with immunohistochemistry and RT-PCR, respectively. The airway inflammation situations were evaluated by histopathology.RESULTS: The airway inflammatory cell infiltration in the DXM-, hemin- and BUD-treated asthma groups was remarkably alleviated compared with that in the asthma model group. Compared with the normal control group, the expression of HO-1 mRNA and protein in lung tissues and the blood COHb content in the asthma model and the DXM-, hemin- and BUD-treated asthma groups were significantly up-regulated. The DXM-, hemin- and BUD-treated asthma groups showed significantly increased expression of HO-1 protein and mRNA in lung tissues compared with the asthma model group. The blood COHb content in the DXM-and the hemin-treated asthma groups was significantly higher than that in the asthma model group.CONCLUSIONS: The expression of HO-1 protein and mRNA in lung tissues and blood HO-1 activity increased in rats with asthma, suggesting that HO-1 may be involved in the pathogenesis of asthma. HO-1 may have a protective effect against the airway inflammation in asthmatic rats. BUD and DXM can up-regulate the expression of HO-1 protein and mRNA, thus providing protective effects against the airway inflammation in asthmatic rats.

OBJECTIVE: To study the effect of extracts of Ginkgo biloba leaf (EGb), a catalyzer of central nervous system, on learning-memory ability and possible mechanism in rats with kindling-induced epilepsy. METHODS: Forty postnatal day 21 (P21) and 40 postnatal day 35 (P35) Sprague-Dawley (SD) rats were randomly respectively assigned to five groups: normal sodium (NS) control, kindling epilepsy model, high, middle and low dosage of EGb-treated kindling epilepsy. The kindling epilepsy model was established by an intraperitoneal injection of pentetrazole (PTZ). The learning-memory ability and NMDA receptor 1 (NMDAR1) expression in the hippocampus were measured by Y-maze test and immunohistochemistry assay respectively. RESULTS: The stimulation times for reaching to academic standard in the Y-maze test in the two ages PTZ kindling groups was significantly more than that in the corresponding NS control groups (P<0.01). After EGb treatment the achievement of the Y-maze test in the three treatment groups was significantly improved in a dose-dependent manner, the higher the dosage, the better the achievement (P<0.01). Immunohistochemistry assay showed that the expression of NMDAR1 in the two ages PTZ kindling groups was significantly higher than that in the corresponding NS control groups (P<0.01). Compared with the corresponding untreated kindling model groups, the expression of NMDAR1 in the two ages EGb treatment groups was significantly reduced in a dose-dependent manner (P<0.01).CONCLUSIONS: EGb can improve learning-memory ability in epileptic rats at different developmental phases in a dose-dependent manner, possibly through a reduction of NMDAR1 expression in the hippocampus.

OBJECTIVE: Calcium plays an important role in the impairment of heart function and arrhythmia under the condition of acute hypoxia, but the mechanism is different from that of chronic hypoxia. This study aimed to evaluate the effect of chronic hypoxia on the expression of calmodulin (CaM) and calcicum/calmodulin-dependent protein kinase II (CaMKII) and the calcium activity in myocardial cells through an animal model of chronic hypoxia in order to get a deeper sight into the mechanism.METHODS: A chronic hypoxia model of the rat was prepared by hypoxia exposure (FiO2 = 10%). The expression of mRNA and protein of CaM and CaMKIIγ and CaMKIIδ in myocardial cells were measured by RT-PCR and Western Blot in normal rats and hypoxia rats 1 and the 3 weeks after exposure. The cardiac cells of the rats from the control group and the 3-week hypoxia group were cultured. Then the intracellular calcium activity was detected using laser confocal equipment. The effect of CaMKII on the calcium activity in myocardial cells was evaluated by the application of KN-62 (CaMKII specific inhibitor). RESULTS: The expression of CaM, CaMKIIγ and CaMKIIδ mRNA in myocardial tissues increased in hypoxia rats compared with that in normal controls (P<0.01). The CaM and CaMKIIδ mRNA expression was different between the 1-week and the 3-week hypoxia groups (P<0.01). The laser confocal demonstrated that the amplitude of calcium wave in hypoxic myocardial cells was not different from that in normal controls, but the duration of calcium wave in hypoxic myocardial cells was longer than that in normal controls (P<0.01). After KN-62 use, the amplitude of calcium wave decreased and the duration of calcium wave prolonged significantly. CONCLUSIONS: The contents of CaM and CaMKII in myocardial cells increased under condition of chronic hypoxia as a compensation to keep calcium homeostasis in a certain time. With more prolonged hypoxia time, abnormal electric activities of heart occurred and the heart function may be impaired.

OBJECTIVE: To study the effect of integrin α2β1 on invasion and migration of SK-N-SH neuroblastoma cells.METHODS: Neuroblastoma SK-N-SH cell line was cultured in the modified eagle's medium. The effects of monoclonal antibodies to integrin α2 and integrin β1 on migration and invasion were measured by inclined test and polycarbonate filters incorporated in modified Transwell chambers respectively．The migration and invasion cells were stained with Gimsa staining and counted under a 200 multiplied microscope. The blocking rate of migration and invasion of cells was calculated.RESULTS: The number of migrated SK-N-SH cells in the anti-α2 and anti-β1 treatment groups (50.9±10.5 and 54.3±9.0 respectively) was significantly less than that in the control group without monoclonal antibody treatment (98.1±7.4) (P<0.01), with a blocking rate of cell migration of 48.1% and 44.5% respectively. The invasion to matrigel of SK-N-SH cells exposed monoclonal antibodies to integrin α2 and integrin β1 was significantly blocked compared with the control SK-N-SH cells, with the number of invasion cells in the anti-α2 and anti-β1 treatment groups of 25.3 ± 4.4 and 18.8 ± 3.9 respectively vs 41.5 ± 4.8 in the control group (P<0.01). The blocking rate of cell invasion in the anti-α2 and anti-β1 treatment groups was 39.0% and 54.7% respectively.CONCLUSIONS: Integrin α2β1 may promote migration and invasion of neuroblastoma cells.