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Objective:To investigate the effects of fluoxetine (Flx) on lipidomics of hippocampal tissue in chronic unpredictable stress (CUS) model rats.Methods:A total of 30 Sprague-Dawley rats were randomly divided into Sham group, CUS group and CUS+ Flx group, with 10 rats in each group. Rats in the CUS group and CUS+ Flx group were received one or two random stimuli every day for 28 days, and then they were received intraperitoneal injection of normal saline(1 ml/kg) and fluoxetine(10 mg/kg) respectively once a day for 14 days. Rats in the Sham group were maintained in their home cages for 28 days, and then received intraperitoneal injection of saline (1 ml/kg) once a day for 14 days. The sugar water preference experiment was carried out 24 hours after the last injection, and then the rats were killed to separate the rat hippocampus. The levels of lipid composition in hippocampus were detected by high performance liquid chromatography-mass spectrometry. The relative content of lipid was analyzed by Simca-p 14.1 and LipidSearch software version 4.1. SPSS 19.0 was used for statistical analysis. One-way ANOVA or Kruskal-Wallis test was used for comparison among groups, and Bonferroni test was used for post-hoc test. Pearson correlation or Spearman correlation was used to analyze the correlation between behavioral indexes and lipid molecular level in hippocampus.Results:There was significant difference in sugar preference test among the three groups ( F=12.830, P<0.001). The percentage of sucrose intake of rats in CUS group ((43.57±12.38)%) was significantly lower than those in Sham group ((67.09±11.81)%) and CUS+ Flx group ((62.74±8.58)%) (both P<0.05). Ninety five differential lipid molecules were screened among the three groups by lipidomic analysis, mainly distributed in glycerophospholipids and sphingolipids. Among them, levels of PE (34∶1e)+ H( r=-0.477), PE(18∶1p/20∶1)+ H( r=-0.433), PE(18∶1/18∶1)+ Na( r=-0.603), PE(36∶2p)-H( r=-0.382), PE(16∶0/20∶4)-H( r=-0.464), PE(18∶0/18.2)-H( r=-0.482), PE(16∶0e/22∶6)-H( r=-0.514), PE(18∶1/20∶4)-H( r=-0.511) and CerG1 (d18∶2/24∶0+ O)+ H( r=-0.490) were negatively correlated with sucrose preference rate (all P<0.05), whereas levels of PE (42∶6p)+ Na( r=0.379), PE(34∶0p)-H( r=0.397) and SM (d22∶1/16.0)+ HCOO( r=0.388) were positively correlated with sucrose preference rate (all P<0.05). Conclusion:Flx improves the depressive-like behavior of CUS model rats, which may be related to the regulation of hippocampal glycerophospholipid and sphingolipid metabolism.
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Objective To study efficacy and safety of continuous renal replacement therapy (CRRT) in the treatment of neonatal sepsis-related acute kidney injury (AKI).Method From June 2011 to June 2018,neonates with sepsis-related AKI hospitalized in the neonatal intensive care unit of our hospital and treated with CRRT were enrolled.Before CRRT,12 h,24 h,48 h after CRRT and by the end of CRRT,their clinical data including renal function,acid-base balance,electrolytes,blood pressure (BP)and the change of hemodynamic indexes were retrospectively analysed.The efficacy and safety of CRRT was evaluated.Kruskal-wallis H test was used for statistical analysis.Result A total of 9 cases of sepsis-related AKI neonates were enrolled in the study,all treated with continuous veno-venous hemofiltration dialysis.5 cases had oliguria,2 cases fluid overload and 2 cases shock.The duration of CRRT was 49 ～ 110 h (76.2 ±23.5) h.12 h after CRRT,BP were maintained at 40 ～60 mmHg and stable during the treatment,the blood pH value increased to 7.35 ～ 7.45 and the oxygenation index reached 200 mmHg.24 h after CRRT,the oxygenation index rose to more than 300 mmHg.Serum potassium,urea nitrogen and creatinine levels decreased significantly after 12 h of CRRT,and reached the normal range after 24 h of CRRT.After 24 h of CRRT,the urine volume significantly increased.Venous catheterization was performed successfully in 9 cases.2 cases had thrombocytopenia,1 case catheterization obstruction and 1 case hypotension during CRRT.No complications such as hypothermia,hemorrhage,thrombosis or infection occurred.All 9 patients were cured and discharged.Conclusion CRRT is safe and effective for the treatment of neonatal sepsis-related AKI.
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Objective To evaluate the safety,feasibility,complications and outcome of continuous renal replacement therapy (CRRT) in neonates weighting less than 3 000 g.Method A total of 6 neonates weighting less than 3 000 g treated with CRRT in the Department of Neonatology,Shanghai Children's hospital,from January 2015 to December 2017 were studied.The birth weight,primary disease,indications of CRRT,treatment duration,age,complications and outcome of the neonates were collected and analyzed.Serum creatinine (Scr),blood urea nitrogen (BUN) and blood ammonia were analyzed before and after CRRT.T test was used for statistical analysis of the data.Result (1) Among the 6 neonates,2 were full-term infants and 4 were premature infants.The average gestational age of the neonates was (35.0± 2.1) weeks and the average birth weight was (2 542±586) g.(2) The catheterization was successful in all of the 6 neonates.The model for CRRT was continuous veno-venous hemofiltration dialysis,and the duration was 50(48,154)h,the neonates' age of CRRT was 3.0(2.0,4.5)days.The primary disease included 3 perinatal asphyxia,1 hemolytic uremic syndrome,1 ornithine transcarboxylase deficiency,1 jejunal atresia.There were 5 patients with acute kidney injury and fluid overload,and another one with hyperammonemia.(3) Compared with before CRRT,serum creatinine,urea nitrogen and serum ammonia all decreased significantly and reached the normal range after CRRT.(4)The complications of CRRT in the 6 neonates included 2 hypotension,1 hypokalemia,1 hypocalcemia and 1 hypophosphatemia.Catheter related infection,blockage and other complications had not occurred.(5) After treatment,3 patients survived,1 witdrew and 2 died.Conclusion The application of CRRT in neonates with weight less than 3 000 g is safe and feasible,the prognosis and survival rate of which can be improved with fewer and controllable complications.
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Objective To investigate the diagnostic value of cerebrospinal fluid protein in the assess-ment of neurological outcome in preterm infants with sepsis. Methods A total of 80 preterm infants with sepsis were enrolled in the department of neonatology of Shanghai Children's Hospital from June 2014 to June 2016. The lumbar puncture was completed within 24 hours after diagnosis of sepsis,and the results of ce-rebrospinal fluid protein were obtained. The prognosis of neurological development was assessed according to Gesell Developmental Quotient ( DQ) at 6 months of adjusted gestational age. DQ> 85 was used as an indi-cator of good prognosis group. DQ≤85 was assigned to the poor prognosis group. The differences in protein content of cerebrospinal fluid between these two groups were retrospectively analyzed. The receiver operating characteristic ( ROC) curve was used to evaluate the diagnostic value of cerebrospinal fluid in evaluating the prognosis of preterm infants with sepsis. Results Cerebrospinal fluid protein content of poor prognosis group was higher than those in good prognosis group[(2005. 56 ± 582. 85)mg/L vs. (1367. 92 ± 362. 29)mg/L, t= -6. 019,P<0. 01]. The area under the ROC curve was 0. 819(95％CI 0. 711 -0. 927,P<0. 05). The optimal threshold of cerebrospinal fluid protein was 1560 mg/L with specificity of 75. 5％ and sensitivity of 81. 5％. Conclusion Cerebrospinal fluid protein content has certain diagnostic value on the assessment of sepsis premature neurological prognosis.
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Objective To explore the clinical features, diagnosis, and treatment of non-immunologic hydrops fetalis (NIHF). Methods The clinical data of 10 cases of NIHF in neonatal intensive case unit during January 2011 to December 2016 were analyzed retrospectively. The related literatures were reviewed. Results In 10 cases of NIHF (6 males and 4 females). the gestational age were 32-42 weeks, and the birth weight was 2.25-3.95 kg. Among them, there were 3 cases of infectious diseases (cytomegalovirus, Streptococcus agalactiae, and parvovirus infection, one case each), 2 cases of fetal cardiovascular abnormalities, 2 cases of chromosomal abnormalities, 1 case of abnormal thoracic structures, 1 case of twin transfusion syndrome, and 1 case of etiology unknown of fetal hydrops. The clinical manifestations showed that there were 8 cases with 2 or more areas of edema (or hydrops), and only 2 cases with skin edema. Finally, 6 cases were cured and discharged, 2 cases were discharged by themself, and 2 cases died. Conclusions Prenatal ultrasound is a reliable method for the diagnosis of NIHF. Fetal edema in early pregnancy, especially with congenital malformations, is recommended for termination of pregnancy. After birth, NIHF should be diagnosed promptly so as to avoid or reduce severe complications.
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Objective To explore the effect of macrolide antibiotics(erythromycin) on tumor necrosis factor(TNF)-α and interleukin(IL)-8 in hyperoxia-induced lung tissue of premature newborn rats,and to study the intervention effect of erythromycin on hyperoxia-induced lung injury.Methods One-day old preterm Sprague Dawley rats were randomly divided into four groups by random number table method:air + sodium chloride group,air + erythromycin group,hyperoxia + sodium chloride group,hyperoxia + erythromycin group.Hyperoxia groups were continuously exposed to oxygen (oxygen ＞ 0.85) and air group in room air.After 1,7,14 days of exposure,the preterm rats of four groups were sacrificed,whole lung of these rats were isolated,the lung histological changes were observed by hematoxylin-eosin staining,TNF-α and IL-8 in pulmonary tissue homogenate were detected by ELISA.Results The results showed that:(1) Compared with air + sodium chloride group,TNF-α and IL-8 expression in hyperoxia + sodium chloride group were significantly increased(P ＜ 0.05) after 1,7 days of exposure [1 d:TNF-α:(16.163 ± 0.574) ng/ml vs.(21.923 ±2.066) ng/ml,IL-8:(18.214 ±3.649) ng/ml vs.(23.546 ± 5.240) ng/ml ;7 d:TNF-α:(15.940 ±0.821) ng/ml vs.(19.688 ±0.764) ng/ml,IL-8:(18.541 ± 4.114) ng/ml vs.(24.255 ±4.692) ng/ml],in particular,TNF-α expression appeared to increase earlier,their expression became significantly weak in 14 days (P ＜ 0.05).(2) Compared with hyperoxia + sodium chloride group,TNF-α and IL-8 expression in hyperoxia +erythromycin group became significantly weak after 1,7,14 days of exposure(P ＜0.05) after the intervention of erythromycin [1 d:TNF-α:(21.923 ± 2.066) ng/ml vs.(18.903 ± 1.851) ng/ml,7 d:IL-8:(24.255 ±4.692) ng/ml vs.(23.508 ±3.543) ng/ml,14 d:TNF-α:(16.443 ±5.466) ng/ml vs.(14.453 ±0.963)ng/ml],but their expression became weaker in 14 days than that in 1,7 days.Conclusion The release of inflammatory mediators TNF-α and IL-8 induced by oxidation outbreak participates in the development of hyperoxia induced lung injury,erythromycin may regulate immune function,inhibits the levels of oxidant-mediated TNF-α and IL-8 induced by oxidation outbreak,and alleviate hyperoxia lung injury in premature rats.
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Objective To explore the efficacy and safety of bedside continuous blood purification (CBP) in the treatment of critically ill neonates.Methods Totally ten critically ill neonates were hospitalized in Department of Neonatal Intensive Care Unit (NICU) in Shanghai Children's Hospital from June 2011 to May 2015, and managed with CBP treatment.The indications of CBP therapy were multiple organ dysfunction syndrome (MODS) failed to conventional treatment or combined with acute renal failure (ARF).The model for CBP was continuous veno-venous hemofiltration dialysis (CVVH).The clinical outcomes included blood electrolytes, serum bio markers, urine output, hemodynamic indicators, dose of intravenous epinephrine before treatment, 6, 12, 24, 48 h after treatment and at the end of CBP.Complications of CBP were also observed.Statistical analysis was performed with ANOVA and Dunnett-t test.Results The underlying problems of the ten newborns were septicemia (n=5), severe neonatal asphyxia (n=2), congenital hereditary metabolic disease (n=2) and traumatic asphyxia (n=l).The venous catheter was successfully inserted for all babies and CBP treatment continued for (86.7 ± 25.9) h averagely with obvious effect.Four of the ten cases were cured and discharged, and the rest six refused to treatment and died after due to irreversible injury of the nervous system although they had survived from the oliguric stage of ARF.The complications of CBP included thrombocytopenia (n=3), catheter blockage (n=2), hypotension (n=l).No hypothermia, thrombosis, bleeding or infection occurred.The mean blood pressure and partial pressure of oxygen in arterial blood/fraction of inspiration oxygen (PaO2/ FiO2) of the ten cases 6 h after the beginning of treatment were higher than those before [(46.4 ± 7.5) vs (36.5 ±8.3) mmHg, 1 mmHg=0.133 kPa;(210.0±62.0) vs (93.0±43.0) mmHg;t=2.647 and 6.378, both P ＜ 0.05].At the 12th hour since treatment start, the blood pH value was 7.4 ± 0.2, which was higher than that before treatment (6.9 ± 0.2, t=2.731, P ＜ 0.05), and kept in normal range.At the 24th hour, the serum levels of potassium, urea nitrogen and creatinine dropped to normal range compared to those before treatment [(4.8±2.9) vs (9.6± 3.6) mmol/L;(7.2±2.3) vs (13.6±6.3) mmol/L;(51.0± 12.0) vs (172.0±23.0) μ mol/L;t=4.571, 5.427 and 21.672, all P ＜ 0.05].Urine output increased from zero before the treatment to (0.7±0.3) ml/(kg · h) after 24 h (t=3.284, P ＜ 0.05).The maintaining dose of intravenous epinephrine decreased since 12 h after the beginning of treatment and was ceased at the 48th hour.Conclusion CBP is an effective and feasible treatment for critically ill neonates.
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Objective To investigate the effect of repetitive transcranial magnetic stimulation (rTMS) on the depressive like behaviors and expression of brain derived neurotrophic factor (BDNF),IL-1β and NF-κB of hippocampal in chronic unpredictable mild stress (CUMS) rats.Methods Thirty-two adult male rats were randomly divided into four groups (n =8):Control group,Control + rTMS group,CUMS group and CUMS + rTMS group.The sucrose preference test,forced swim test and open field test were used to evaluate depressive like behaviors for each groups.In addition,the expression of BDNF,NF-κB and IL-1 β in hippocampal were detected by western blot and ELISA after behavioral test,respectively.Results 1.The effects of rTMS on depressive like behaviors of CUMS rats:in the sucrose preference test,the sucrose preference rate of CUMS rats (0.67 ± 0.06) was significantly lower than Control group (0.91 ± 0.04),which was higher in the CUMS + rTMS group (0.83 ±0.08).In the forced swim test,the immobility time of CUMS group ((26.88 ± 11.33) s) was longer than Control group ((15.22 ± 6.75) s) and CUMS + rTMS group ((18.41 ± 6.95) s).In the open field test,both the total distance travelled and number of central area entry times of CUMS group((849.165 ± 769.01) cm,(7.42 ± 5.68))were significantly shorter ((6224.81 ± 1403.2) cm) and smaller (22.86 ± 3.72) than Control group,and those of the CUMS + rTMS were longer ((4105.57 ± 1516.92)cm) and larger (21.25 ± 3.45).All the behavioral results were statistically significant (P＜ 0.05).And of all the aforementioned behavioral parameters,there were no significant differences between Control group and Control + rTMS group(P＞0.05).2.The effects of rTMS on the hippocampal expressions of BDNF,NF-κB and IL-1β in CUMS rats:compared with Control group,the hippocampal expression of BDNF in CUMS rats was significantly decreased,while the expressions of NF-κB and IL-1β in the hippocampus were significantly increased (P＜ 0.05).Compared with CUMS group,the hippocampa expression of BD-NF in the CUMS + rTMS group was increased,and the expressions of NF-κB and IL-1β in the hippocampus was significantly decreased (P ＜ 0.05).The expressions of BDNF,NF-κB and IL-1β had no differences between Control group and Control + rTMS group.Conclusion rTMS increased the expression of BDNF,reduced the production of NF-κB and IL-1β,and alleviated depressive like behaviors in CUMS rats.
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Objective To investigate the effect of quetiapine on the behavior and expression of pERK1/2 in chronic unpredictable stress(CUS) model rats.Methods 32 adult male Sprague Dawley rats were randomly divided into four groups (n =8 for each group):control group,CUS group,CUS + QUE (5 mg/kg,L) group and CUS + QUE(10 mg/kg,M)group.The rats in control group were left undisturbed in their home cage for 28 days and the other groups were exposed to 28 consecutive days of CUS,then the rats in control group and CUS group were treated with 1％ DMSO in saline (5 ml/kg,intraperitoneal injection),the rats in CUS + QUE (L)group and CUS + QUE(M) group respectively treated with quetiapine (5 mg/kg)or quetiapine(10 mg/kg) for consecutive 7 days.The weight data of each group were recorded,and the behavioral changes in these rats were analyzed by open field test and forced swimming test;and the expression of pERK1/2 was measured by Western blot.Results (1)Compared with control group,quetiapine (10 mg/kg) ameliorated the inhibition of body weight gain that induced by chronic unpredictable stress (P ＜ 0.05),but quetiapine (5 mg/kg) did not have this effect.(2)Open field and Forced swimming test showed significant difference (P ＜ 0.05) of horizontal motion distance (F =17.846),the number of central region entering(F=4.720) and the immobility time(F=26.090) in each group.And these tests showed that horizontal motion distance and the number of central region entering in CUS group ((6696.30 ±1061.19)mm,(19.63 ±9.15)times) were significantly lower than that of control group ((10824.61 ± 1399.37) mm,(37.75 ± 13.02) times) and CUS + QUE (M) group ((9637.51 ± 1630.16) mm,(32.38 ± 6.23)),while the immobility time (110.73 ± 15.98)s were significantly higher than that of control group((66.13 ± 5.18)s)and CUS + QUE (M) group((73.40 ± 11.99) s,P ＜ 0.05).But there was no significant difference between that of CUS group and CUS + QUE(L) group(P＞0.05).(3)The expression of pERK1/2 in CUS group showed significant decrease when compared with control group or CUS + QUE (M) group,but showed no significant difference with CUS + QUE(L) group(F=6.641,P＜ 0.01).Conclusion Quetiapine can ameliorate depressive-like behaviors induced by chronic unpredictable stress,and this effect may be carried out by up-regulation the expression of pERK1/2 in the hippocampus.
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Objective To investigate the effect of rosmarinic acid on the behavioral changes in enhanced single prolonged stress (ESPS) model rats and the levels of interlukin-1β (IL-1β) and interlukin-6 (IL-6) in the hippocampus.Methods 48 adult male Sprague Dawley rats were randomly divided into six groups (n =8):Control group,Control + RA (L) group,Control + RA (H) group,ESPS group,ESPS + RA (L) group and ESPS + RA (H) group.Behavioral changes of these rats were analyzed by open field test and elevated plus-maze.The levels of IL-1 β and IL-6 were measured by enzyme linked immunosorbent assay (ELISA).Results (1) Open field test showed that the number of central region entering and the fraction of time exploring in center of ESPS group were significantly reduced than that of Control group ((18.13 ± 10.15) times,(26.68 ± 10.06) ％) and ESPS + RA (H) group ((16.88 ± 8.81) times,(25.08 ± 8.52) ％) (P ＜ 0.05).And it showed no significant difference among Control + RA(L) group,Control + RA(H) group and Control group.Meanwhile,there was also no statistic difference between ESPS group and ESPS + RA(L) group.(2) Elevated plus-maze test showed that percentages of open arm entries and fraction of time exploring in open arm in reference to total number of entries into all arms and total time spent on all arms in ESPS group were significantly reduced than that of Control group((37.38 ± 8.24)％,(17.63 ±4.74)％) and ESPS + RA(H) group((33.72 ±9.49)％,(16.99 ±4.28)％) (P ＜ 0.05),but there was no significant difference between that of ESPS group and ESPS + RA(L) group(P＞0.05).It also showed no significant difference among Control + RA (L) group,Control + RA (H) group and Control group.(3) Compared with ESPS group,RA(10mg/kg) reduced the levels of IL-1β and IL-6 in the hippocampus,but RA(5mg/kg) did not have this effect.(4) Correlation analysis results showed the level of IL-1β in the hippocampus was negatively related with the ameliorated PTSD-like behaviors of ESPS exposure rats.Conclusion RA can ameliorate PTSDlike behaviors of ESPS exposure rats,and this effect may be carried out by down-regulating the levels of IL-1β and IL-6 in the hippocampus,especially the IL-1β.
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ObjectiveTo investigate the effects of ziprasidone on the behavior and the expression of pERK1/2 in posttraumatic stress disorder(PTSD) model rats.Methods 24 adult male SD rats weighing (200 ±20) g were randomly divided into four groups (n =6):control group,single prolonged stress and foot shock (SPS&S) group,ziprasidone group and ziprasidone + U0126 group.The fear response to environment,high alertness,and anxiety & depression behavior of rats were tested by the open field,elevated plus-maze,and the expression of pERK1/2 was measured by Western blot.ResultsIn open field test(OFT),the SPS&S group( (76.23 ± 54.76) cm for horizontal motion distance,(4.60 ± 1.14) for the number of entering central region) showed significant difference compared with control group ( (343.77 ± 74.22 ) cm,( 12.40 ± 3.36 ) ) or ziprasidone group ( ( 274.98± 83.56) cm,( 12.00 ± 2.92) ) (P ＜ 0.01 ),but showed no significant difference with ziprasidone + U0126 group ( ( 138.14 ± 41.98) cm,(5.00 ± 1.58) ) (P ＞ 0.05 ).The results of elevated plus maze (EPM) were in accordance with the results of OFT.The expression of pERK1/2 in SPS&S group and ziprasidone + U0126 group showed significant decrease when compared with control group or ziprasidone group (P ＜ 0.01 ).ConclusionZiprasidone can obviously improve fear response to environment,high alterness and anxiety & depression behavior of rats,and these effects of ziprasidone may be carried out by up-regulation the expression of pERK1/2.
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ObjectiveTo explore the relation of heart rate variability (HRV) in neonatal asphyxia with myocardial injury.MethodsContinuous electrocardiographic monitoring by 24-hour Holter recordings was performed in 53 neonates with asphyxia and 40 healthy newborn.The difference of HRV with sinus rhythm was analyzed.Time-domain indexs included standard deviation of R-R intervals (SDNN) ; standard deviation of all mean 5-minute R-R intervals (SDANN) ;standard deviation of all R-R intervals for all 5-minute segments of 24 hours (SDNNindex ) ;root mean squared successive difference (rMSSD) ;percent of NN50 in the total number R-R intervals ( PNN50 ).Results( 1 ) Maximum heart rate,minimum heart rate and average heart rate of 24-hour Holter in healthy newborn were faster than those in newborn with neonatal asphyxia ( P ＜ 0.05 ).And the heart rate was faster in newborn with mild neonatal asphyxia than that in newborn with serious neonatal asphyxia ( P ＜ 0.05 ).(2) SDNN,SDANN of HRV index analysis showed significantly difference between healthy newborn and asphyxia newhom ( P ＜ 0.05 ).There were no difference of SDNN,SDANN,SDNNindex,rMSSD and PNN50 between mild and serious neonatal asphyxia (P ＞ 0.05 ).No significant differences of SDNNindex,rMSSD and PNN50 were found among three groups.ConclusionMyocardial injury caused by neonatal asphyxia can lead to damage of cardiac autonomic nevous and affect heart rate changes.The degree of myocardial injury is related to the degree of neonatal asphyxia.
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Objective To investigate the changes of N-terminal pro-brain natriuretic peptide (NT-proBNP) in neonates with hypoxic-ischemic encephalopathy (HIE) complicated by myocardial ischemic injury. Methods Forty neonates with HIE ( 16 cases with concurrent myocardial injury and 24 cases without) were enrolled. Twenty healthy neonates were used as control. Plasma NT-proBNP levels were measured using enzyme immunoassay. Cardiac function was measured by echocardiography. Results ( 1 ) The mean plasma NT-proBNP levels in patients with myocardial injury[(350 ± 56) pmol/L]were significantly higher than those in patients without myocardial injury[(135 ± 37 ) pmol/L]and in the control group [(117 ±23) pmol/L](P ＜0. 05). (2) The NT-proBNP levels in mild,moderate and severe HIE neonates were ( 132 ±34) pmol/L, (247 ±43) pmol/L and (343 ±53) pmol/L. Compared with the control group,the NT-proBNP levels in the neonates with moderate and severe HIE significantly increased. There were significant differences in the NT-proBNP levels among the mild, moderate and severe HIE neonates ( P ＜ 0. 05 ).(3) In patients with myocardial injury,the NT-proBNP levels significantly decreased in the convalescent phase [(250±78) pmol/L]compared with those in the acute phase[(350±56) pmol/L](P ＜0.05). (4) The NT-proBNP levels were significantly related with left ventricular ejection fraction. Conclusion Plasma NT-proBNP levels increase in neonates with HIE complicated with myocardial ischemic injury in the acute phase.Detection of NT-proBNP levels maybe useful in the diagnosis of myocardial ischemic injury and severe HIE.
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BACKGROUND: A great quantity of cell loss in early stage following stem cell transplantation can significantly affect transplantation effect. Presently, it is confirmed that overexpression of AKT1 gene significantly inhibit cell apoptosis. OBJECTIVE: To explore whether AKT1 gene overexpression can block stem cell apoptosis under hypoxic condition following pig autologous bone marrow mesenchymal stem cell (BMSC) transplantation, and the effect of repairing damaged myocardium. DESIGN, TIME AND SETTING: The randomized controlled animal study was performed at the Soochow University from August 2005 to February 2007.MATERIALS: A total of 24 healthy male Meishan pigs were supplied by the Animal Experimental Center of Soochow University. METHODS: The CDS (regulation domin of AKT1) AKT1-cDNA fragment was amplified. Lentivector Packaging Kit was used to transfect BMSCs after synthesized with pCDH1-AKT1 shuttling plasmid. Following BrdU labeling, models of myocardial infarction were constructed by occluding the distal left anterior descending coronary artery in pigs with gelatin sponge. 4 weeks later, pigs were randomly divided into four groups: the model control group, the DMEM group, the BMSCs group, and the AKT-transfected group. In model control group, there was no other injection after occluding the left anterior descending coronary artery. In the DMEM group, 5 mL DMEM was injected into the coronary artery. 5 mL BMSCs (1×10~7 cells) were infused into the coronary artery in the BMSCs group. 5 mL BMSCs transfected with the AKT1 gene were injected in the AKT-transfected group MAIN OUTCOME MEASURES: Western blot analysis and real time RT-PCR were used to test the plasmid. The cardiac function was evaluated by magnetic resonance image. Histological characteristics of the myocardium were observed using immunohistochemistry. Serum vascular endothelial growth factor and transforming growth factor β1 levels were determined by ELISA. RESULTS: AKT1-cDNA was cloned into pCDH1-MCS1-EF1-copGFP and the sequence was confirmed in comparison with the published one. AKT mRNA expression could be detected distinctly 24 and 48 hours after transfecting cells. The expression of AKT1 intensity in MSCs remained strong 2 weeks later with detected by real time RT-PCR and Western blot analysis. AKT1-mRNA transcriptional levels were 120 times of primary cells. Before the cell implantation, the left ventricular end-diastolic dimension increased and the stroke volume decreased in the myocardial infarction hearts. The cardiac function was significantly improved after cell implantation, and the implanted MSCs prevented the infarct region from thinning and expanding, improved contraction and increased perfusion in all groups relative to the control hearts. The left ventricular chamber size was smaller in the hearts with being transplanted cells than that in the control hearts. Moreover, the improvement was even markedly greater in AKT-transfected group (P < 0.05). Hematoxylin-eosin staining results showed that fibering was significant in the model control group and DMEM group. Island-like myocardium was observed in the infarct zone of the BMSCs group and AKT-transfected group, and plenty of small vessels-shape structure was detected in the AKT-transfected group. Immunohistochemistry demonstrated that Von Willebrand Factor (vWF) and Cx-43 expression was determined in the myocardium in the BMSCs group and AKT-transfected group, and the proportion of BrdU and Cx-43-positive cells to BrdU-positive cells was significantly greater in the AKT-transfected group compared with the BMSCs group 4 weeks following transplantation (P < 0.05). Following cell transplantation, vascular endothelial growth factor levels were gradually increased, peaked at 1 week, gradually decreased, and reached a normal level at 4 weeks. Transforming growth factor p1 levels were gradually reduced, and significantly less than the model control group, DMEM group 4 weeks later (P < 0.05), and significantly lower than that pretransplantation (P < 0.05).CONCLUSION: Using lentiviral vector to construct with AKT1 gene could stably make BMSCs overexpress AKT1. The BMSCs engraftment in host myocardium might improve the left ventricle function by attenuating the contractile dysfunction and pathologic thinning in this model of left ventricular wall infarction. AKT1 overexpression can significantly improve cardiac function following infarction.