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Effect of NADPH oxidase on hypoxia-inducible factor-1alpha and endothelin-1 expression in human umbilical endothelial cells and its possible mechanism.

Lan DONG ; Ke-Zheng CHEN ; Liu-Fang HE

Chinese Journal of Contemporary Pediatrics.2006;8(4):323-326.

OBJECTIVETo study the effect of NADPH oxidase on hypoxia-inducible factor (HIF)-1alpha and endothelin (ET)-1 expression in human umbilical endothelia cells (HUVECs) and its possible mechanism.

METHODSTwenty-five bottles of HUVECs culture fluid were randomly assigned into five groups: group A (normoxic control), group B (hypoxic), group C (NADPH oxidase inhibitor apocynin + normoxic), group D (H2O2 which can degrade HIF-1alpha rapidly+hypoxic) and group E (H2O2+apocynin+normoxic), with five bottles in each group. The culture supernates were collected and the total protein was extracted 3 hrs after treatment. Western Blot and ELISA were used to detect the HIF-1alpha protein expression in HUVECs and the ET-1 level in the culture supernates respectively.

RESULTSThere was a lower expression of HIF-1alpha protein (0.336 +/- 0.012) and lower ET-1 levels (5.87 +/- 2.22 pg/mL) in group A. The HIF-1alpha protein expression in groups B and C (0.773 +/- 0.018 and 0.888 +/- 0.022) and ET-1 levels (95.38 +/- 8.06 and 33.67 +/- 4.21 pg/mL) were noticeably higher than in group A (P < 0.05). The groups D and E had the HIF-1alpha protein expression levels similar to group A, but the ET-1 levels in group D (108.43 +/- 8.38 pg/mL) and group E (109.66 +/- 5.80 pg/mL) were significantly higher than in group A (P < 0.05).

CONCLUSIONSHypoxia or apocynin can increase the HIF-1alpha and ET-1 expression in HUVECs. H2O2 can inhibit the HIF-1alpha expression but increase the ET-1levels. It is speculated that NADPH oxidase as an oxygen sensor regulates the HIF-1alpha expression by changing the intracellular redox reaction and that except HIF-1, H2O2 might contribute to ET-1 synthesis and release.

Blotting, Western ; Cell Hypoxia ; Cells, Cultured ; Endothelial Cells ; metabolism ; Endothelin-1 ; analysis ; genetics ; metabolism ; Enzyme-Linked Immunosorbent Assay ; Humans ; Hypoxia-Inducible Factor 1, alpha Subunit ; analysis ; metabolism ; NADPH Oxidases ; physiology

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Effects of L-arginine on the expression of insulin-like growth factors and insulin-like growth factor binding protein 3 in rats with intrauterine growth retardation.

Yan LU ; Xiao-Mei LIU ; Shu-Qin LI

Chinese Journal of Contemporary Pediatrics.2006;8(4):319-322.

OBJECTIVEIntrauterine growth retardation (IUGR) may contribute to the disorder of development of fetal brains. L-arginine has been known to be effective in blood vessel distension and improving the blood circulation of placentas. Recent studies have shown that L-arginine can ameliorate the placental hypoxia and improve the development of fetus. This study aimed to explore the effects of L-arginine on the expression of insulin-like growth factor (IGF)-I, IGF-II, IGF binding protein-3(IGFBP3)and IGF-I mRNA in brains of IUGR rats and the possible mechanisms of L-arginine.

METHODSThirty-six pregnant rats were randomly assigned into four groups: Control, Model, Low dose L-arginine (100 mg/kg) and High-dose L-arginine (200 mg/kg L-arginine) groups (n=9 each). IUGR was induced by passive smoking in rats from the last three groups. L-arginine was administered for the last two groups between days 8 and 20 of gestation. On day 21 of gestation, the pup rats were delivered by cesarean section. The levels of IGF-I, IGF-II and IGFBP3 in the brains of pup rats were measured by enzyme-linked immunoadsordent assay (ELISA) and the expression of IGF-I mRNA was detected by fluorescence quantitative PCR (FQ-PCR).

RESULTSThe levels of IGF-I, IGF-II and IGF-I mRNA expression in the Model group were significantly lower than in the Control group, with the IGF-I levels of 0.789 +/- 0.062 ng/mg vs 0.947 +/- 0.042 ng/mg, the IGF-II levels of 0.270 +/- 0.020 ng/mg vs 0.374 +/- 0.015 ng/mg and the IGF-I mRNA expression of (13.12 +/- 1.39) x 10(4) cps/mug RNA vs (21.28 +/- 3.54) x 10(4) cps/mug RNA (P < 0.01). In contrast, the IGFBP3 levels in the Model group were significantly higher than in the Control group (0.253 +/- 0.011 ng/mg vs 0.089 +/- 0.015 ng/mg; P < 0.01). Low or high dose L-arginine treatment increased significantly the IGF-I levels from 0.789 +/- 0.062 ng/mg (Model group) to 0.937 +/- 0.067 ng/mg (low dose group) or 0.858 +/- 0.077 ng/mg (high dose group), the IGF-II levels from 0.270 +/- 0.020 ng/mg (Model group) to 0.318 +/- 0.018 ng/mg (low dose group) or 0.354 +/- 0.021 ng/mg (high dose group) and the IGF-I mRNA expression from (13.12 +/- 1.39) x 10(4) cps/mug RNA (Model group) to (19.24 +/- 2.48) x 10(4) cps/mug RNA (low dose group) or (17.35 +/- 2.30) x 10(4) cps/mug RNA (high dose group) (P < 0.01). The IGFBP3 levels were significantly reduced after low or high dose L-arginine treatment (0.132 +/- 0.006 ng/mg or 0.146 +/- 0.009 ng/mg) compared with those of the Model group (0.253 +/- 0.011 ng/mg) ( P < 0.01).

CONCLUSIONSL-arginine can increase the levels of IGF-I and IGF-II and the IGF-I mRNA expression, and decrease the IGFBP3 level in the brain of rats with IUGR induced by passive smoking, thereby offering protective effects against IUGR.

Animals ; Arginine ; pharmacology ; therapeutic use ; Female ; Fetal Growth Retardation ; metabolism ; prevention & control ; Insulin-Like Growth Factor Binding Protein 3 ; analysis ; Insulin-Like Growth Factor I ; analysis ; genetics ; Insulin-Like Growth Factor II ; analysis ; Male ; RNA, Messenger ; analysis ; Rats ; Rats, Wistar

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Neuroprotective effect of hypothermia on hypoxic-ischemic brain injury in mice.

Xiao-Yang WANG ; Chang-Lian ZHU ; Fa-Lin XU ; Xiu-Yong CHENG ; Lin QIU ; Sheng-Hai HU

Chinese Journal of Contemporary Pediatrics.2006;8(4):315-318.

OBJECTIVEThe study was to investigate the effect of different temperatures during hypoxia on brain injury in mice of different ages.

METHODSNewborn C57/BL6 mice at 7 days or 21 days of life were subjected to left carotid artery ligation followed by exposure with 10% oxygen. The mice were kept in a incubator with a predetermined, constant temperature, either 34 degrees centigrade (Hypothermia group) or 36 degrees centigrade (Normothermia group). Brain injury was evaluated 7 days after hypoxia-ischemia (HI). Active caspase-3 and apoptosis-inducing factor (AIF) expressions in the brain tissue were detected by immunohistochemistry and Western Blot was used to evaluate the phosphor-Akt (P-Akt) expression in the brain tissue at 24 hrs post-HI.

RESULTSBrain injuries, including the cortex, hippocampus, striatum and thalamus injuries, occurred in the Normothermia group at 7 days post-HI. The brain cortex showed cystic cavitation in the postnatal day (P)7 pups mice and laminar infarct of the brain cortex was observed in P21 mice. In the Hypothermia group, the P7 mice did not present with laminar infarct of the cortex and had lower scores of neuropathological lesions in cortex, hippocampus, striatum and thalamus than P7 mice from the Normothermia group (P < 0.01); the cortex injuries were significantly relieved but the injuries of hippocampus, striatum and thalamus in P21 mice were similar to those from the Normothermia group. Active caspase-3 (7.0 +/- 5.6) and AIF positive cells (3.7 +/- 6.2) in the cortex of P7 mice from the Hypothermia group were significantly lower than those of the Normothermia group (51.5 +/- 23.2 and 31.8 +/- 22.4) at 24 hrs post-HI (P < 0.01). Wetstern Blot showed the P-Akt expression was obviously decreased in the ipsilateral hemisphere to the occlusion compared with that of the contralateral hemisphere after HI in the Normothermia group (P < 0.05), while in the Hypothermia group the P-Akt expression was not significantly different between the two hemispheres.

CONCLUSIONSHypothermia has protective effects against HI insults. The protection was more pronounced for the immature brain than the mature brain.

Active Transport, Cell Nucleus ; Age Factors ; Animals ; Apoptosis Inducing Factor ; metabolism ; Brain ; pathology ; Caspase 3 ; Caspases ; metabolism ; Hypothermia, Induced ; Hypoxia-Ischemia, Brain ; metabolism ; pathology ; therapy ; Mice ; Mice, Inbred C57BL ; Proto-Oncogene Proteins c-akt ; metabolism

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McCune-Albright syndrome: a difficult and complicated case study.

Li-Yang LIANG ; Zhe MENG ; Qiao-Hui ZENG ; Wen-Yi LI

Chinese Journal of Contemporary Pediatrics.2006;8(4):311-314.

McCune-Albright syndrome is a rare G proteins alpha disorder. The disorder is characterized by polyostotic fibrous dysplasia, sexual precocity and hyperpigmented macules. It is caused due to mutations in the gene Gsalpha that incodes the alpha subunit of the trimeric guanosine triphate-binding protein. There is no specific treatment for this syndrome. Treatment is generally symptomatic. This paper reported three cases of McCune-Albright syndrome and reviewed the relevant literatures regarding to the pathogenesis, pathological features, diagnosis and treatment. All three cases presented with a characteristic triad: polyostotic fibrous dysplasia, sexual precocity and hyperpigmented macules and were thus definitely diagnosed with McCune-Albright syndrome.
Child ; Child, Preschool ; Diagnosis, Differential ; Female ; Fibrous Dysplasia, Polyostotic ; diagnosis ; etiology ; pathology ; therapy ; Humans ; Prognosis

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Levels of IL-12 produced by dendritic cells and changes of TH1/TH2 balance in children with Henoch-Schonlein purpura.

Da-Qing SUN ; Qiu-Ye ZHANG ; Zeng-Yi DONG ; Feng BAI

Chinese Journal of Contemporary Pediatrics.2006;8(4):307-310.

OBJECTIVETo study the changes of IL-12 produced by dendritic cells in peripheral blood in children with Henoch-Schonlein purpura (HSP), and to explore its influence on TH1/TH2 balance in order to elucidate its significance in the pathogenesis of HSP.

METHODSThe levels of interferon-gamma (IFN-gamma), interleukin-4 (IL-4) and interleukin-12 (IL-12) in plasma were determined by ELISA in 60 HSP children (HSP group) and 21 healthy children (Control group). Peripheral blood mononuclear cells (PBMC) of 22 HSP patients and 21 healthy children were cultured in vitro and then were transformed into dendritic cells. The levels of IL-12 in the supernatant were detected by ELISA and the positive expression rate of CD1a(+) was detected by indirect immunofluorescence procedure.

RESULTS1) The levels of IFN-gamma and the ratio of IFN-gamma/IL-4 in plasma of the HSP group were lower than those of the Control group (IFN-gamma 30.59 +/- 11.27 pg/mL vs 43.38 +/- 19.19 pg/mL; IFN-gamma/IL-4 ratio 0.70 +/- 0.28 vs 1.33 +/- 0.57) (P < 0.01). The levels of IL-12 in the HSP group were also lower than those of the Control group (153.95 +/- 91.88 pg/mL vs 323.06 +/- 162.34 pg/mL; P < 0.01). In contrast, the levels of IL-4 were higher than those of the Control group (45.08 +/- 9.19 pg/mL vs 32.95 +/- 7.10 pg/mL; P < 0.01). The plasma levels of IL-12 positively correlated with the IFN-gamma levels (r=0.52, P < 0.01) and the ratio of IFN-gamma/IL-4 (r=0.43, P < 0.01) in the HSP group. 2) The IL-12 levels in the supernatant of the HSP group were lower than those of the Control group (357.06 +/- 153.56 pg/mL vs 489.80 +/- 213.45 pg/mL; P < 0.05), and had a positive correlation with the plasma IL-12 levels (r=0.74, P < 0.01). 3) The positive expression rate of CD1a(+) of the HSP group was lower than that of the Control group [(27.42 +/- 10.75)% vs (35.68 +/- 12.18)%; P < 0.05], and positively correlated with the IL-12 levels in the supernatants (r=0.57, P < 0.01) and in plasma (r=0.68, P < 0.01).

CONCLUSIONSThere was an imbalance of TH1/TH2 in HSP children. The decrease of TH1 function had a positive correlation with the low levels of IL-12 in plasma, while the latter correlated closely with decreased number and / or function of dendritic cells, suggesting that the decreased number and / or function of dendritic cells in peripheral blood resulted in the imbalance of TH1/TH2 indirectly.

Adolescent ; Antigens, CD1 ; analysis ; Cells, Cultured ; Child ; Child, Preschool ; Dendritic Cells ; immunology ; Female ; Humans ; Interferon-gamma ; blood ; Interleukin-12 ; biosynthesis ; blood ; Interleukin-4 ; blood ; Male ; Purpura, Schoenlein-Henoch ; immunology ; Th1 Cells ; immunology ; Th2 Cells ; immunology

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Variation and significance of serum and stool IL-18 and IFN-gamma levels in children with rotavirus enteritis.

You-Gui GAO ; Yu JIN ; Yi-Ling LIU ; Xin-Hua YE

Chinese Journal of Contemporary Pediatrics.2006;8(4):304-306.

OBJECTIVETo study the variation and significance of serum and stool IL-18 and IFN-gamma levels in children with rotavirus enteritis.

METHODSSerum and stool specimens from 50 children with acute rotavirus enteritis were collected before treatment. Serum and stool levels of IL-18 and IFN-gamma were measured using ELISA. Serum and stool specimens from 21 age and gender-matched healthy children were used as the Control group.

RESULTSSerum and stool levels of IL-18 and IFN-gamma in patients with rotavirus enteritis were significantly higher than those in the Control group. There was a negative correlation between the serum IFN-gamma level and the frequency of vomiting (r=-0.368, P < 0.05). The stool IL-18 level negatively correlated to the frequency of diarrhea (r=-0.414, P < 0.05). A positive correlation was found between the serum levels of IL-18 and IFN-gamma (r=0.416, P < 0.05).

CONCLUSIONSSerum and stool levels of IL-18 and IFN-gamma were increased and associated with the severity in children with rotavirus enteritis. IL-18 and IFN-gamma might have protective effects against acute rotavirus infection at the early stage.

Child, Preschool ; Enteritis ; immunology ; Feces ; chemistry ; Female ; Humans ; Infant ; Interferon-gamma ; blood ; physiology ; Interleukin-18 ; blood ; physiology ; Male ; Rotavirus Infections ; immunology

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Efficacy of intravenous or atomizing ambroxol for prevention of respiratory distress syndrome in preterm infants.

Qin HU ; Jin-Mei LIAN ; Jian-Qing LI

Chinese Journal of Contemporary Pediatrics.2006;8(4):301-303.

OBJECTIVEAmbroxol induces the synthesis of surfactant in lung alveolar type II cells. Some studies have shown its effectiveness for the prevention of respiratory distress syndrome (RDS) in preterm infants. This study aimed to compare the efficacy of two different ways of ambroxol administration, ie, intravenous injection and atomizing inhalation, for the prevention of RDS in preterm infants.

METHODSA total of 125 preterm infants born between 28-37 weeks of gestation were randomly assigned into three groups: Intravenous and Atomizing ambroxol treatment groups (n=40 each) or Control group (n=45). The Intravenous group was injected with 15 mg/kg of ambroxol through the umbilical vein immediately after birth and then received 30 mg/kg of ambroxol daily for 2 days by intravenous drip. The Atomizing group was administered with 30 mg/kg of ambroxol daily for 2 days by atomizing inhalation immediately after birth. The Control group received no ambroxol treatment. The incidences of RDS and complications as well as the blood gas results 6 hrs after birth were compared among the three groups.

RESULTSThe incidence of RDS was 7.5%, 5.0% and 24.4% in the Intravenous, Atomizing and Control groups respectively. There were no significant differences in the incidence of RDS between the two ambroxol treatment groups. However, the incidence of RDS in the two treatment groups were noticeably lower than in the Control group (P < 0.05). The blood gas results did not show significant differences between the two ambroxol treatment groups but both groups demonstrated improved blood gas results compared with the Control group at 6 hrs after birth (P < 0.05). The incidence of complications, such as pulmonary hemorrhage, respiratory failure, intraranial hemorrhage, in the two ambroxol treatment groups was reduced compared with the Control group (P < 0.05), but there were no differences between the two ambroxol groups.

CONCLUSIONSEarly administration of either intravenous or atomizing ambroxol can produce a positive efficacy for the prevention of RDS in preterm infants. The two different ways of administration seem to result in a similar efficacy in the prevention of RDS.

Administration, Inhalation ; Ambroxol ; administration & dosage ; adverse effects ; pharmacology ; Female ; Humans ; Infant, Newborn ; Infant, Premature ; Injections, Intravenous ; Male ; Pulmonary Surfactants ; metabolism ; Respiratory Distress Syndrome, Newborn ; prevention & control

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Characteristics of community-acquired Methicillin-resistant Staphylococcus aureus infection in children.

Ying-Hui HU ; Jing-Hui ZHEN ; De-Huan ZHAO

Chinese Journal of Contemporary Pediatrics.2006;8(4):298-300.

OBJECTIVETo investigate the prevalence and drug resistant patterns of community-acquired Methicillin-resistant Staphylococcus aureus (MRSA) in children.

METHODSSamples of sputum, blood, liquor puris/secretion of skin or stool in Beijing Children's Hospital between January, 2002 and March, 2005 were cultured. The characteristics of community-acquired MRSA infection were analyzed and compared with hospital-acquired MRSA infection.

RESULTSA total of 25 strains of MRSA were found during the study period and they accounted for 4.7% in 512 strains of Staphylococcus aureus. Of the 25 strains of MRSA, 20 strains were community-acquired but only 5 were hospital-acquired. The prevalence of MRSA infection in Staphyloccus aureus has kept rising over last two years, 3.1% in 2003, 5.4% in 2004 and 7.2% in the first season of 2005. There were no statistical differences in the results of antimicrobial susceptibility testing and multi-resistance testing between the groups of community-acquired and hospital-acquired MRSA. In both groups, all isolates were susceptible to vancomycin. The percentage of the patients with underlying disease in the hospital-acquired infection group was significantly higher than in community-acquired infection group (P < 0.05), but the onset age was not different.

CONCLUSIONSThe prevalence of community-acquired MRSA infection tends to increase in children. The drug resistant patterns of community-acquired MRSA were not significantly different from the hospital-acquired MRSA in children.

Child ; Child, Preschool ; Community-Acquired Infections ; drug therapy ; microbiology ; Humans ; Infant ; Infant, Newborn ; Methicillin Resistance ; Staphylococcal Infections ; drug therapy ; microbiology ; Staphylococcus aureus ; drug effects

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Effects of salbutamo and ipraopium bromide inhalation on pulmonary function in young children with asthmatoid bronchitis.

Wen-Hui JIANG ; Li DENG ; Hui-Hong WEN ; Jia-Lu YU ; Qiang ZENG

Chinese Journal of Contemporary Pediatrics.2006;8(4):295-297.

OBJECTIVEThe efficacy of bronchodilator in asthmatoid bronchitis remains controversial. This study was designed to investigate the effects of bronchodilators, salbutamo and ipraopium bromide, on the pulmonary function in young children with this disease.

METHODSPulmonary function tests were performed in 20 children with asthmatoid bronchitis (2 months-2.5 years of age) before and 30, 60, and 120 minutes after salbutamo and ipratropium bromide inhalation. The indexes of pulmonary function measured included tidal breathing flow volume (TBFV) loop, percent of tidal volume to peak tidal expiratory flow (%V-PF), terminal flows per peak expiratory flow (25/PF), peak tidal expiratory flow (PTEF), rate of mid-expiratory to mid-inspiratory flow (ME/MI), respiratory rate (RR) and tidal volume per kilogram (TV/kg).

RESULTSBefore drug inhalation, the descending branch of the TBFV loop was depressed. The PTEF shifted forward and %V-PF (0.19 +/- 0.04) and 25/PF (0.42 +/- 0.11) decreased. These changes did not improve and the remaining indexes, RR, ME/MI and TV/kg, 30, 60, and 120 minutes after drug inhalation also remained similar to before inhalation.

CONCLUSIONSSalbutamo and ipratropium bromide inhalation did not improve the airway resistance and ventilation function in children with asthmatoid bronchitis. This suggests that the efficacy of bronchodilator in the treatment of this disease is doubtful.

Administration, Inhalation ; Albuterol ; administration & dosage ; Asthma ; drug therapy ; physiopathology ; Bronchitis ; drug therapy ; physiopathology ; Bronchodilator Agents ; administration & dosage ; Child, Preschool ; Female ; Humans ; Infant ; Ipratropium ; administration & dosage ; Lung ; drug effects ; physiopathology ; Male

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Value of carboxyhemoglobin in the diagnosis of neonatal jaundice.

Xiu-Hong PU ; Qian LI ; Xiao-Qing GUO ; Tao AN ; Meng QIU ; Xiao-Qing WANG

Chinese Journal of Contemporary Pediatrics.2006;8(4):291-294.

OBJECTIVETo study the value of carboxyhemoglobin (COHb) in the diagnosis of neonatal jaundice.

METHODSThis study consisted of 189 patients with neonatal jaundice due to hemolytic disease (n=75), infectious disease (n=52), intracranial hemorrhage (n=32) and breast-milk feeding (n=30). One hundred and forty-two neonates without pathological jaundice that were gestational age, postnatal age- and birth weight-matched were used as the Control group. The level of arterial capillary blood COHb was detected by a 270 CO-oximeter connected to an 800 series system. Total serum bilirubin (STB) content was measured using an Abbott Spectrum CCX chemistry analyzer. The levels of COHb and STB were measured at baseline, and again in patients with jaundice due to hemolytic disease after intravenous gammaglobulin treatment for 2 days.

RESULTSThe levels of COHb [(3.64 +/- 0.83)%] and STB (330.84 +/- 77.15 micromol/L) in patients with jaundice due to hemolytic disease were significantly higher than those measured in the Control group [COHb (2.38 +/- 0.35) %; STB 130.18 +/- 32.86 micromol/L] (P < 0.01). The levels in patients with jaundice due to intracranial hemorrhage were also significantly higher than those in the Control group [COHb (2.48 +/- 0.53) % vs (2.24 +/- 0.32) %; STB 184.15 +/- 29.35 micromol/L vs 112.11 +/- 17.45 micromol/L; P < 0.05]. The patients with jaundice due to infectious disease or breast-milk feeding only demonstrated higher levels of serum STB (P < 0.01) while COHb levels were not different compared with the Control group. The patients with jaundice due to hemolytic disease or intracranial hemorrhage presented with hemolytic unconjugated hyperbilirubinemia and had significantly higher COHb levels and lower STB levels than those patients with nonhemolytic unconjugated hyperbilirubinemia (caused by breast jaundice) (P < 0.01). The levels of COHb [(2.68 +/- 0.51) %] and STB (230.18 +/- 42.96 micromol/L) in patients with jaundice due to hemolytic disease decreased markedly after intravenous gammaglobulin treatment (P < 0.01).

CONCLUSIONSThe COHb level can be used as a supplementary indicator of increased bilirubin production. The elevation of COHb can be useful in the diagnosis of neonatal jaundice since COHb is elevated in hemolytic disease and intracranial hemorrhage.

ABO Blood-Group System ; immunology ; Bilirubin ; blood ; Carboxyhemoglobin ; analysis ; Female ; Humans ; Immunoglobulins, Intravenous ; therapeutic use ; Infant, Newborn ; Jaundice, Neonatal ; diagnosis ; therapy ; Male

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