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Abstract:OBJECTIVE: To investigate possible mechanisms resulting in pulmonary hypertension (PH) in patients with congenital heart disease (CHD). METHODS: Thromboxane B2/6keto Prostaglandin F1a (TXB2/6-K-PGF1a), Von Willebrand factor (VWF: Ag) and tissue plasminogen activator (tPA) plasma levels were measured. Changes in pulmonary vascular endothelial cells were assessed with light and electron microscopy. Ninety-nine children with CHD were divided into 3 groups according to pulmonary vascular resistance (PVR) (Group Ⅰ: PVR≤3.5 wood's unit; Group Ⅱ: PVR 3.5～5 wood's unit; Group Ⅲ: PVR>5 wood's unit). RESULTS: TXB2/6-K-PGF1a and VWF:Ag increased significantly in PH, and varied in different PH groups (P<0.05). There was a positive association between PVR and VWF: Ag (r=0.89, P<0.05) and between pulmonary artery resistance (PAR)and VWF: Ag (r=0.82, P<0.05). The levels of tPA in PH did not differ from normal. With the progressive changes on the light microscopy, the increases in the volume density of rough endoplasmic recticulum and microfilament bundles on the transmission electron microscopy were more significant and the immunostain for VWF: Ag was more intense. CONCLUSIONS: The functional and structural changes of endothelial cells are important in the pathogenesis of PH secondary to CHD. The experiments provide a theoretical basis for the modification of PH secondary to CHD with drug therapy.

Abstract:OBJECTIVE: To assess micturition characteristics, bladder capacity and plasma and urine osmolarity in neonates. METHODS: Fortyfive neonates were divided into three groups according to post natal ages (Group A=～3 d, Group B=～7 d, Group C=>7 d). The time of micturition and voided volume were recorded. The residual urine volume was detected using a bladder volume instrument (BladderScan BVI 2500). Plasma and urine osmolarity were measured using the FM-6 pattern freezing point osmometer. RESULTS: ① There were significant differences (P<0.05) in the daynight voided volume and voided frequency between Group A and Groups B and C. ② The bladder emptying rates, maxium bladder capacity(MBC) and plasma urine osmolarity of Groups A, B and C were 0.80±0.07, 0.82±0.09, 0.79±0.10; 56±6, 65±14, 61±15 (ml); 285±5, 287±7, 286±8 (mOsm/L); 202±59, 185±65, 216±92 (mOsm/L) respectively and there was no significant difference between the 3 groups. ③ The multiple stepwise regression method was used to estabilish the best regression equation for predicting the MBC, which was 42±0.17X (ml) ［X=8 hours (10 Am～6 Pm) voided volume, F=15.561 (P<0.01)］. CONCLUSIONS: The bladder of normal neonates empties almost completely and its function is stable. There is no difference in the renal concentration function of neonates between day and night.

Abstract:OBJECTIVE: To study the changes in Nacety-β-D-amino glucosidase (NAG) and urinary retinal binding protein (RBP) observed after renal function impairment in asphyxiated newborns. METHODS: Colorimetry of p nitrophoenol and ELISA were adopted to detect the activity of NAG and the content of RBP in asphyxiated full term and premature infants on days 1, 3 and 7, and in matched nonasphyxiated control infants. RESULTS: The levels of NAG and RBP were higher in both asphyxiated full term and premature infants than in the control group (P<0.01). The changes were most significant on day 3 after birth, and correlated with the degree of asphyxia. The degree and duration of renal dysfunction also correlated significantly with the degree of asphyxia. Urinary RBP levels in the asphyxiated newborns rose significantly within 1 day after birth compared to the controls (P<0.01), while there was no significant difference in the levels of NAG on day 1 of life in asphyxiated infants (P>0.05). CONCLUSIONS: NAG and RBP appear to be indexes of early diagnosis in renal dysfunction in asphyxiated infants, and RBP is a more sensitive one.

Abstract:OBJECTIVE: To study the changes of serum procalcitonin (PCT) in neonates with sepsis. METHODS: Serum PCT was determined using an immunoradiometric assay in 20 newborn infants with neonatal sepsis, 24 neonates with hypoxicischemic encephalopathy (HIE) and 16 healthy newborn controls. RESULTS: Increased levels of serum PCT were found in neonates with sepsis compared with the control group ［(112.23±10.13) μg/L vs (8.65±2.14) μg/L］, (P<0.01). After 1 week of treatment with appropriate antibiotics, PCT levels returned to normal. There was no difference between HIE neonates and the control group. CONCLUSIONS: Elevated serum PCT levels are noted in neonatal sepsis. PCT might be of value in the diagnosis of neonatal sepsis.

Abstract:OBJECTIVE: To explore the relationship between cytomegalovirus (CMV), herpes simplex virus (HSV) and human parvovirus B19 (HPV-B19) and fetal malformation. METHODS: There were 16 infants with congenital malformation in the malformation group and 25 normal infants in the control group. CMV, HSV and HPV-B19 infections were diagnosed using the polymerase chain reaction technique (PCR) in the main organs and placentas of the 2 groups. RESULTS: The infection rate with CMV and HSV in the malformation group was 50.0% and 37.5% respectively, compared with the 8.0% and 8.0% respectively in the control group. (P<0.05). The HPV-B19 infection rate didn't differ in the 2 groups. The brain, liver and lungs were the organs most commonly involved in CMV infection, whereas in HSV infection the liver was most commonly involved, with HPV-B19 most commonly affecting the spleen. CONCLUSIONS: CMV and HSV infections are closely related to congenital malformation, but HPV B19 infection is not related to malformation.

Abstract:OBJECTIVE: To study the dynamic changes in renal Ca2+ ATPase after acute intrauterine ischemia in fetal rats,and to explore the underlying mechanism of intracelluar Ca2+ overload secondary to perinatal asphyxia.METHODS: A rat model of acute intrauterine ischemia was developed using different degrees of clamping of the vesseles supplying blood to the uterus. A graded reperfusion model was made by reperfusing the uterine vessels for different lengths of time after clamping. The activity of Ca2+ATPase in cell membranes and mitochondrial fractions in fetal rat kidneys was assayed biochemically.RESULTS: Ca2+ATPase activity in cell membranes and mitochondria in the shamoperated group was (7.476±0.353) and (3.470±0.270) μmol.Pi/mgprot.h respectively. Activities in both fractions decreased after 15 minutes of ischemia ［(6.411±0.210) and (2.886±0.245) μmol.Pi/mgprot.h respectively; P<0.01］ and continued to decrease after 45 minutes ［(5.772±0.177) and (2.500±0.282) μmol.Pi/mgprot.h respectively; P<0.05］. During the reperfusion stage (after 15 minutes of ischemia), activities continued to decrease reaching a nadir after 8 hours ［(5.513±0.197) and (2.411±0.197) μmol.Pi/mgprot.h respectively］. There was significant difference between the 8hour reperfusion group and the 15-minute ischemia group (P<0.01). After 15 h of reperfusion enzyme activity levels began to increase. By 24 hours, the activity of Ca2+ATPase in cell membranes had increased to normal levels, but the actvity in mitochondria was still low compared with the 15-minute ischemia group ［(3.234±0.143) and (2.886±0.245) μmol.Pi/mgprot.h respectively; P<0.05］. CONCLUSIONS: Ca2+ATPase activity decreases in the fetal kidney after acute intrauterine ischemia. Dynamic changes in Ca2+ATPase activity may explain the intracellular Ca2+ overload leading to hypoxic ischemic renal injury.

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Abstract:OBJECTIVE: To investigate the role of endogenous nitric oxide (NO) in hyperoxiainduced lung injury, by studying the effect of hyperoxia on NO synthesis and nitric oxide synthase (NOS) expression in the lung of preterm rats. METHODS: Threedayold preterm rats were randomly assigned to a hyperoxia group (90% oxygen) and room air group. After 3 days or 7 days of exposure, the ratio of lung wet weight/dry weight (W/D), lung morphometry, NO content in bronchoalveolar lavage fluid (BALF), and the cellular distribution and expression of endothelial NOS (eNOS) and inducible NOS (iNOS) in the lung were measured. RESULTS: After 3 days of exposure, when compared with the control group, the hyperoxia group showed acute lung injury characterized by the presence of hyperaemia, red cell extravasation, and inflammatory infiltration. After 7 days of exposure, W/D was also increased in the hyperoxia group compared to the controls (5.54±0.41 VS 5.00±0.15, P<0.05), in addition to the pathologic changes. After 3 and 7 days of exposure, the NO content in BALF was significantly elevated in the hyperoxia group compared with that of the air group ［(17.06±5.86) μmol/L vs (5.59±2.03) μmol/L and (23.75±4.07) μmol/L vs (7.93±2.33) μmol/L, respectively, P<0.01］. In the lungs of the hyperoxia group, immunostaining for iNOS was observed in the airway and alveolar epithelium, and in inflammatory cells, to a greater degree than in the air group. The expression of iNOS in rats was stronger after 7 days of hyperoxic exposure than after 3 days. After 7 days of exposure, stronger immunostaining for eNOS in the airway epithelium of the hyperoxia group than of the air group was seen. CONCLUSIONS: Hyperoxia can significantly upregulate the expression of iNOS and eNOS in inflammatory cells and epithelium in the lungs of preterm rats and thus promote NO generation, suggesting that endogenous NO may mediate hyperoxiainduced pulmonary injury.

Abstract:OBJECTIVE: To study the effect of MK-801, the antagonist of NMDA receptor, on Caspase-3 activation and apoptosis after cerebral hypoxic ischemic injury in neonatal rats. METHODS: Sevendayold rat pups were injected with either 0.5 mg/kg MK-801 or normal saline immediately after cerebral hypoxicischemic injury (HI). The pups were killed 24 h after the injection. Brain damage was evaluated using MAP-2 immunostaining. Caspase-3 activation was examined using antip17 subunit antibody. Apoptosis was examined by in situ labeling of hairpin probe (HPP). The infarction area was calculated according to MAP-2 staining. Active Caspase-3 cells and HPP positive cells were counted in the MAP-2 negative area of the cortex. RESULTS: The infarction area was reduced from (52±12)% to (23±5)% after the treatment with MK-801. A few Caspase-3 and HPP positive cells were also found in the cortex of normal rat brains. The number of positive cells reached a peak at 24 h after HI. Both the number of Caspase-3 (from 65±8 to 40±6.7 per high power visual field) and HPP (from 61.6±11.5 to 12.6±5.2 per high power visual field) positive cells were decreased after the treatment with MK-801. CONCLUSIONS: MK-801 inhibits Caspase-3 activation and reduces the number of apoptotic cells. NMDA is involved in the activation of the apoptotic process in the immature brain after HI.

Abstract:OBJECTIVE: To investigate the mechanism of apoptosis in cultured rat islets. METHODS: Rat islets were sampled for isolation, 1, 3, 7, 14, 21 day (s) in culture. The distribution of reticulin for peri insular basement membrane was observed and stained by Foot's assay. Apoptotic cells were assessed by electron microscopy and fluorescent microscopy with Hoechst 33258 staining. The islet viability was determined by MTT colorimetric assay indirectly. The positive rate of islet Fas and Fas ligand (Fas L) protein expression was measured by immunocytochemistry ABC staining. RESULTS: Immediately after islet isolation, the reticulin periinsular basement membrane was absent. Typical morphological characteristics of islet apoptosis were observed, such as nuclear shrinkage and migration of chromatin towards the nuclear membrane, after 3 d of culture. The apoptotic rate was (7.6±5.8)% after 3 d, and increased to (63.0±2.6)% and (47.2±8.1)% after 14 d and 21 d of culture. Catenin β expression diminished after 7 d of culture by immunocytochemistry ABC staining. The positive rate of islet Fas protein expression increased from (8.1±1.8) after 3 d to (38.5±4.7)% after 14 d, and (35.6±6.5)% after 21 d of culture. Although the expression of Fas-L protein was negative before days 3, the positive rate of Fas-L protein was (6.8±3.2)% after 7 d, and was upregulated significantly after 14 d and 21 d of culture to (19.6±4.8)% and (12.4±7.1)% respectively. Moreover, cumulative quantities of insulin production and islet viability were reduced significantly. CONCLUSIONS: Rat islets are primed to undergo apoptosis in culture, and this event involves some association between cell-surface Fas and its ligand.

Abstract:OBJECTIVE: To study the relationship between calmodulin (CaM) concentrations in brain tissues of newborn rats and the pathogenesis of hypoxicischemic encephalopathy (HIE) and the therapeutic effect of cerebrolysin on HIE. METHODS: Morphological and metabolic effects of cerebral ischemia were investigated in a newborn rat model of neonatal HIE. In addition, the effect of cerebrolysin (EBEWE, Austria) on acute brain hypoxicischemia was studied. Eighty eight HIE newborn rats (48 cerebrolysintreated and 40 controls) were subjected to left cerebral artery ligation and one hour later they were placed in a hypoxic environment (8% oxygen and 92% nitrogen mixed) for two hours. Immediately afterwards, cerebrolysin (2.5 ml/kg body mass) was administered daily, either by intracerebroventricular infusion (i.c.v) or by intramuscular injection. CaM concentrations were determined using cyclicnucleotide dependent phosphodiesterase methods, and 15 normal rats were used as the control group. RESULTS: CaM concentrations in brain tissues increased in HIE group at different duration after hypoxicischemia: 24 h=(521.27±46.04) μg/g brain tissue; 48 h=(509.52±35.98) μg/g brain tissue; 72 h=(421.05±31.81) μg/g brain tissue, compared to the control group ［(187.63±54.22) μg/g brain tissue］ (P<0.01). Cerebrolysin treatment groups facilitated recovery, as indicated by a decrease in CaM concentration (i.c.v group: 24 h=(435.21±56.17) μg/g brain tissue; 48 h=(260.38±32.43) μg/g brain tissue; 72 h=(197.64±19.21) μg/g brain tissue; i.m.group: 24 h=(441.04±30.66) μg/g brain tissue; 48 h=(305.39±32.99) μg/g brain tissue; 72 h=(217.71±52.89) μg/g brain tissue (P<0.01). Morphological examination showed that the volume of neuron degeneration, necrosis and intercellular edema was increased in HIE groups and significantly reduced in cerebrolysin treatment groups. CONCLUSIONS: CaM concentration in brain tissues is increased after hypoxicischemic injury and may be implicated in the pathogenesis of newborn rats with HIE. Cerebrolysin is effective in protecting brain cells from further damage through its influence on cerebral CaM concentration.

Abstract:OBJECTIVE: To observe the effect of hypoxia on the activity of the tissue-type plasminogen activator (TPA) of brain microvascular endothelial cells in mice. METHODS: Microvascular endothelial cells obtained from the brains of newborn mice were cultured in primary and passage under room air and hypoxic conditions to observe the variation in TPA activities. TPA activity was assayed using the enzyme linked immunosorbent assay (ELISA) technique. RESULTS: Following hypoxia, TPA activity in endothelial cells increased in primary culture compared with the controls: (22.5±1.5)×10-2 IU/ml vs (19.4±1.7)×10-2 IU/ml (P<0.01). TPA activity following hypoxia was not altered in passaged cells. CONCLUSIONS: Neuronal damage following hypoxia may be associated with increased endothelial TPA activity.

Abstract:OBJECTIVE: To explore the effect of angelica polysaccharide (AP) on the thymocyte proliferation and RBC C3bR immune adhesion function in radiated mice. METHODS: Mice were divided into control, irradiated and AP treated also irradiated groups (10 per group). The proliferation of thymocytes and RBC C3bR rosette formation rate were then examined on day 10 after AP was given. RESULTS: The thymocyte proliferation responses (0.24±0.01) and the RBC C3bR rosette formation rate ［(16.46±0.76)%］ of the irradiated control group were significantly lower than those of the non irradiated controls ［(0.35±0.01) and (22.30±1.26)%, respectively; P<0.01］］. The thymocyte proliferation (0.31±0.01) and RBC 36bR rosette formation ［(21.07±1.22)%］ in the treatment group were significantly higher (P<0.01) than those of the irradiated control group, and approached nonirradiated control levels. CONCLUSIONS: AP can reverse radiationinduced reduction in the proliferation response of thymocytes and in RBC immune adhesion function.

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Abstract:OBJECTIVE: To explore the mechanism of ameliorative effects of the nerve growth factor (NGF) on newborn rats with hypoxic ischemic brain damage (HIBD). METHODS: NGF dosage (0.1 μg/kg and 0.3 μg/kg) was administrated intraperitoneally to the rat pups 30 minutes before and immediately following the hypoxic exposure. The percentage of brain water content was determined according to the formula: (wet weight dry weight)/wet weight×100=% water content. Brain tissue calcium concentration was measured with atomic absorption spectroscopy. Monoamine neurotransmitter concentration in the rat brain was measured using high performance liquid chromatography with electrochemical detector (HPLCECD). RESULTS: Brain water content, calcium conentration and monoamine neurotransmitters (except DOPAC) concentration increased significantly after hypoxicischemic insult in the ipsilateral hemisphere of the HIBD group compared with the contralateral hemisphere and/or control group: water contents=(91.44±2.21)% vs (82.99±6.53)% (P<0.05) and (91.44±2.21)% vs (87.22±1.13)% (P<0.05) respectively; calcium concentration=(255.24±122.6) μmol/g dry weight vs (44.12±13.54) μmol/g dry weight (P<0.01) and (255.24±122.6) μmol/g dry weight vs (42.68±4.35) μmol/g dry weight (P<0.01), respectively. There were significant discreases in water content, calcium concentration and monoamine neurotransmitters (except DOPAC) concentration in the 0.1 μg/kg and 0.3 μg/kg NGFtreated HIBD group (compared with untreated HIBD group): water contents=(86.86±0.86)% and (87.45±0.3)%, respectively; calcium concentration=(44.34±3.10) μmol/g dry weight and (42.03±14.99) μmol/g dry weight, respectively. No differene was found between the NGF group and control group. CONCLUSIONS: Water content, calcium concentration and monoamine neurotransmitters may be involved, alone or interactively, in the pathophysiologic process of HIBD. NGF has ameliorative effects on newborn rats with HIBD, possibly through maintaining calcium homeostasis, ameliorating brain edema and attenuating or inhibiting the neurotoxic effects of monoamine neurotransmitters.

Abstract:OBJECTIVE: To investigate nosocomial infection risk factors in acute leukemia in children in order that they are wellrecognized by clinic doctors and the mortality rate associated with them will be decreased. METHODS: A retrospective review of the medical records of 306 children with acute leukemia admitted between January, 1995 and December, 2000 was performed. The data were analyzed with the single factor χ2 test, t test and multifactor logisticregression analysis. RESULTS: The incidence of nosocomial infection was 28.43%(87/306), and consisted mainly of respiratory tract and oral infections. The length of hospitalization, white blood cell count, neutrophil and platelet count were found to be independent risk factors for nosocomial infection. CONCLUSIONS: The length of hospitalization, reduced white blood cell count and neutrophil and platelet count are independent risk factors for nosocomial infection in children with leukecnia.

Abstract:OBJECTIVE: To study the expression of CD4+ (helper T lymphocyte), CD45RA+ and CD45RO+ in PBMCs of infants with RSV LRTIs. METHODS: CD4+ and CD45RA+ were detected in 30 patients with RSV LRTIs and CD45RO+ in PBMCs was also assayed except for CD+4 and CD45RO+ in 11 of them with the flow cytometric analysis. Nine healthy children were used as the controls. RESULTS: In RSV infants, CD4+ cells ［(32.74±10.60)%］ decreased and CD45RA+ ［(80.65±10.35)%］ increased compared with the controls ［(40.76±6.82)% and (67.34±7.54)%, respectively］ (P<0.01). CD45RO+ cells also decreased, but not significantly. CONCLUSIONS: There is a disorder of immune function in PBMCs in infants with RSV.

Abstract:OBJECTIVE: To explore changes of the serum granulocyte colony-stimulating factor (G-CSF) in children with acute respiratory airway infection. METHODS: Venous blood was obtained from 30 normal children, 189 children with acute respiratory airway infection without treatment and 50 children (from the group of 189 children) treated with antibiotics for 5～7 days. G-CSF levels were determined using the double antibody sandwich enzymelinked immunosorbent assay (PAS ELISA) and C reaction protein (CRP) levels were assayed using singleradial immunodiffusion. RESULTS: Of the 189 patients with acute respiratory airway infection, 105 were positive for G-CSF and 68 were positive for CRP (>12 ug/ml). Both G-CSF and CRP were positive in 23.8% of the patients. After a 5～7 day treatment course, the CRP of 50 turned from positive to negative and the G-CSF of 16 remained positive. CONCLUSIONS: Serum G-CSF levels increase in the stage of acute infection.

Abstract:OBJECTIVE: To explore the effect of cardiopumlonary bypass using the bloodless priming technique in open-heart surgery in children. METHODS: Fifty cases of atrial septal defect (ASD) or ventricular atrial septal (VSD) were randomly divided into a bloodless priming group (n=25) and a control group receiving blood priming (n=25). The Hct, furosemide dosage, urine output, pericardial and mediastinal drainage, and time of wakening and of mechanical ventilatory support were deternined intraoperatively and postoperatively. RESULTS: The intra and 12 h post operative Hct ［(19.0±6.8) % and (30.4±25.2)%, respectively］ in the bloodless group was lower than that in the blood primed group ［(23.4±10.6)% and (33.2±23.4)%, respectively］ (P<0.05). The intra and post operative doses of furosemide ［(5.2±0.8) mg and (4.5±0.6) mg, respectively］ in the bloodless group were larger than those in the blood primed group ［(1.2±1.0) mg and (1.5±0.5) mg, respectively］ (P<0.05). The intra and 24 h post operative urine output in the blood less group ［(218±56) ml and (278±38) ml, respectively］ was higher than that in the bloodprimed control group ［(78±36) ml and (189±62) ml, respectively］. The Hct did not differ between the bloodless group and bloodprimed group 24 h after the operation ［(38.6±25.2) % vs (38.8±24.3) %］. Pericardial and mediastinal drainage and time of wakening and postoperative mechanical ventilatory support did not differ in the two groups. CONCLUSIONS: Cardiopulmonary bypass using the bloodless priming tecnique appears to be safe and practicable in pediatric openheart surgery procedures.

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