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Objective To determine the dynamic changes in adenosine monophosphate-activated protein kinase (AMPK) in neurons of neonatal rats suffering from hypoxic ischemic brain injury.Methods Twenty-four-hour old and seven-day old neonatal rats were used in this study.A classic primary cortical neuron oxygen glucose deprivation (OGD) model and neonatal rat hypoxic ischemic encephalopathy (HIE) model were employed.Lactic dehydrogenase (LDH) and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) were used to evaluate neuron viability and damage.The expression of phosphorylated adenosine monophosphate-activated protein kinase (P-AMPK),phosphorylated activated protein kinase (P-Akt) and Cleaved Caspase-3 in neurons and brain tissue was measured by Western blot at different time points after OGD or HIE.The Student-t test was used for statistical analysis.Results (1) Compared with the control group,LDH levels at 2,4,8 and 24 h after OGD were higher (all P＜0.05) and optical absorption levels of MTT were lower (all P＜0.05).(2) Levels of P-AMPK in the OGD group were higher than those in the control group,and showed a time-dependent increase at 30 min and 2,4,8 and 24 h (all P＜0.05).The expression levels of P-AMPK in the HIE group were higher than those in the control group (0.345 ± 0.038,0.387 ± 0.112 and 0.618 ± 0.075 at 1,3 and 7 days after HIE,and 0.132±0.032 in the control group,all P＜0.05).(3) The levels of P-Akt increased above the control levels at 30 min (0.991 ±0.134 vs 0.304±0.050),reached a maximum level at 2 h (1.183± 0.107),and then gradually declined,whereas the levels of Cleaved Caspase-3 started to increase at 30 min,and remained elevated at 24 h (all P＜0.05).Conclusion Following hypoxic ischemic brain damage,the expression of P-AMPK is significantly increased in both in vivo and in vitro studies in a time-dependent manner.

7.Efficacy comparison of aminophylline combined with naloxone and caffeine citrate in prevention of apnea of prematurity

Wenbin LI ; Liwen CHANG ; Wei LIU ; Zhihui RONG ; Baohuan CAI

Chinese Journal of Applied Clinical Pediatrics 2014;29(18):1381-1384

Objective To investigate the efficacy and safety of aminophylline,caffeine citrate and aminophylline combined with naloxone in prevention of apnea of prematurity(AOP).Methods Ninety-four infants with a birth weight ＜ 1 500 g and gestational age ＜ 34 weeks admitted to Department of Pediatrics,Tongji Hospital Affiliated to Tongji Medical College,Huazhong University of Science and Technology between Jan.2010 and Jan.2012 were randomly divided into 3 groups.(1) Aminophylline group (n =30):30 infants received a loading dose of 4-5 mg/kg of aminophylline and then maintained by a dose of 2 mg/kg,with intravenous drip q12 h.(2) Caffeine citrate group(n =32):a loading dose of 20 mg/kg of caffeine citrate was followed by a daily maintained dose of 5 mg/kg,with intravenous drip per day.(3) Aminophylline combined naloxone group (observation group,n =32):32 infants were treated with Aminophylline combined with naloxone.After 6 hours of the first dose of aminophylline,a dose of 0.1 mg/kg naloxone was injected,q12 h.Then the two drugs were used alternately.The mortality and incidence of AOP,bronchopulmonary dysplasia(BPD),retinopathy of prematurity (ROP) and brain injury were evaluated,and drug-related side effects were recorded.Results 1.There was no significant difference in gender,gestational age,birth weight,maternal antenatal glucocorticoid application,pregnancy (including multiple pregnancy) and delivery,5 min Apgar score,oxygen therapy,and the application of positive airway pressure as well as pulmonary surfactant among the 3 groups(all P ＞0.05).2.Compared with aminophylline group,the incidence of apnea of caffeine group and observation group were significantly lower (F =6.704,P ＜ 0.05),but there was no significant difference between caffeine group and observation group (P ＞0.05).3.There was no statistically significant difference in mortality,duration of oxygen therapy,the incidence of ROP,brain injury and hearing loss,postmenstrual age,body weight at discharge,the duration and cost of hospitalization among the 3 groups(all P ＞0.05).4.The BPD incidence in caffeine group[9.4％ (3/32 cases)] and observation group [12.5％ (4/32 cases)] were lower than that in Aminophylline group [20.0％ (6/30 cases)],but there was no statistical significance among the 3 groups(P ＞ 0.05).5.No drug-related side effects were recorded in the 3 groups.Conclusions It is safe and effective to use aminophylline combined with naloxone in prevention of AOP,and its efficiency is similar to caffeine citrate.