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Objective:To explore the value of video-electroencephalography (VEEG) combined with amplitude-integrated electroencephalography (aEEG) in evaluating the condition and prognosis of neonatal hyperbilirubinemia brain injuries.Methods:A total of 120 children with hyperbilirubinemia treatedin the Central Hospital of Enshi Tujia and Miao Autonomous Prefecture from July 2019 to July 2020 were enrolled. According to MRI with or without T 1 weighted imging (T 1WI) hyperintensity changes in the globus pallidus, they were divided into the brain injury group (52 cases) and the normal group(68 cases). According to the severity of brain injury, the brain injury group was divided into bilirubin encephalopathy group (23 cases) and subclinical bilirubin brain injury group (29 cases). According to the scores of Gesell Development Scale, the brain injury group was divided into good prognosis group (37 cases)and poor prognosis group (15 cases). The diagnostic value in brain injury with hyperbilirubinemia, the evaluation of the severity of brain injury and the predictive value of VEEG and aEEG were analyzed. Results:The abnormal rates of VEEG and aEEG in the brain injury group were higher than those in the normal group: 76.92% (40/52) vs. 8.82% (6/68), 80.77% (42/52) vs. 11.76% (8/68), the differences were statistically significant ( χ2 = 57.81 and 57.73, P<0.01). The abnormal rates of VEEG and aEEG in bilirubin encephalopathy group were higher than those in subclinical bilirubin brain injury group: 91.30% (21/23) vs. 65.52% (19/29), 95.65% (22/23) vs. 68.97% (20/29), the differences were statistically significant ( χ2 = 4.80 and 5.88, P<0.05). There was no significant difference in abnormal rates of VEEG and aEEG between the good prognosis group and poor prognosis group ( P>0.05). The results of operating characteristic curve analysis showed that the areas under the curve of VEEG combined with aEEG in the diagnosis of brain injury with hyperbilirubinemia, evaluation of the severity of brain injury, predicting the prognosis of children were higher than those of each examination method used alone ( P<0.05). Conclusions:VEEG combined with aEEG has diagnostic value for neonatal brain injury with hyperbilirubinemia, and has evaluation value for severity and prognosis of the disease.
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Objective To study the regulatory role of microRNA-16 (miR-16) on human pulmonary surfactant associated protein (SP).Method Human alveolar epithelial A549 cells were transfected by miR-16 analogue,analogue negative control,inhibitor and inhibitor negative control.Blank control group was also set up at the same time.The proliferative abilities of the cells in each group were measured using cell counting kit-8 (CCK8) test.The expressions of miR-16,SP-A,SP-B and SP-C mRNA were examined using reverse transcription polymerase chain reaction (RT-PCR).The protein levels of SP-A,SP-B and SP-C were examined using western blotting method.Result RT-PCR showed that the expression of miR-16 after transfected with miR-16 analogue (34.11± 1.79) was higher than the negative control group (1.65 ± 1.07) and the blank control group (1.07 ±0.50).The expression of miR-16 after transfected with miR-16 inhibitor (0.36±0.05) was lower than the negative control group (0.96±0.13) and the blank control group (1.05±0.20).The differences were significant (all P＜0.05),and indicated that miR-16 over-expression and suppression were successfully achieved.Compared with the blank control group,cell proliferation at different time points in the analogue negative control group and the inhibitor negative control group showed no significant differences (all P＞0.05).Compared with the blank control group (1.02±0.19,1.01±0.09,1.01± 0.12) and the analogue negative control group (1.08±0.24,1.00±0.14,1.00±0.05),miR-16 down-regulated the mRNA expressions of SP-A,SP-B and SP-C (0.58±0.16,0.67±0.05,0.61±0.12).On the other hand,compared with the blank control group (1.02±0.19,1.01±0.09,1.01±0.12) and the inhibitor negative control group (1.05±0.22,0.99±0.13,0.98±0.10),miR-16 up-regulated the mRNA expressions of SP-A,SP-B and SP-C (1.66±0.33,1.29±0.11,1.23±0.12)(all P＜0.05).The trends of protein level of SP-A,SP-B and SP-C were related to their mRNA expression.Conclusion This study indicates that miR-16 inhibits pulmonary surfactant associated protein in A549 cells.
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Objective To investigate the incidence and risk factors of neonatal asphyxia in Hubei Enshi Tujia and Miao Autonomous Prefecture. Methods Live born infants, including those diagnosed with neonatal asphyxia, were recruited from 16 different hospitals in Hubei Enshi Tujia and Miao Autonomous Prefecture from January to December of 2016. The 16 hospitals included four grade A tertiary hospitals (three general hospitals and one traditional Chinese medicine hospital) and 12 grade A secondary hospitals (eight general hospitals, one maternal and child health hospital and three traditional Chinese medicine hospitals). A retrospective investigation was conducted using questionnaire to analyze the basic information, perinatal risk factors and prognosis of those infants. Chi-square test was used for statistical analysis. Results Among 22 294 recruited live born infants, 733 (3.29%) were diagnosed with neonatal asphyxia on discharge, including 627 (85.54%) mild cases and 106 (14.46%) severe cases. And neonatal asphyxia resulted in deaths of 27 cases (3.68%). The risk factors for neonatal asphyxia included multiple pregnancy, pregnancy conceived with assisted reproductive technology, premature infant, low birth weight infant, fetal malposition, congenital malformation, male infant, born during transfer, mother of Tujia nationality, low educational level (primary school or lower), living in rural area, the number of antenatal visits ≤3, history of early threatened abortion, anemia in pregnancy, hypertensive disorders of pregnancy, chorioamnionitis, abnormal pregnancy history and abnormality of umbilical cord, amniotic fluid or placenta. Conclusions The incidence of neonatal asphyxia in Enshi area is obviously higher than the national average. The main risk factors for neonatal asphyxia in this area are related to maternal background and the living condition of the mother during pregnancy, delivery as well as the newborn at birth.
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Objective To explore the role of miR-20a on pulmonary surfactant synthesis of alveolar epithelial cells A549 and its potential mechanism.Methods Lentivirus miR-20a overexpression vector(miR-20a group) or lentivirus no-load vector(no-load group) was transfected into A549 cells,and the expression of green fluorescent protein(GFP) was observed to determinate the transfection effficiency;cell proliferation was detected by using 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT);the bioinformatics software and database were applied to predict and analyze the target genes of miR-20a about lung development;expressions of miR-20a,pulmonary surfactant-associated protein A(SP-A),pulmonary surfactant-associated protein B(SP-B),pulmonary surfactant-associated protein C(SP-C) and pulmonary surfactant-associated protein D(SP-D) mRNA were detected by using quantitative real-time PCR(qPCR);the expressions of SP-A protein,SP-B protein,SP-C protein,SP-D protein and protein signal transducers and activators of transcription 3 (STAT3) were detected by using Western blot.Results Observation of GFP expression under a fluorescent microscope indicated similar transfection efficiency,and real time-PCR showed that the expression of miR-20a increased after being transfected with lentivirus miR-20a overexpression vector(3.85 ± 0.18)compared with the normal group (0.99 ± 0.04)and the no-load group (1.21 ± 0.12),and the differences were significant(t =10.85,9.64,all P ＜0.001).As a result,lentivirus miR-20a overexpression vector was constructed successfully.Online software predicted that STAT3 gene was likely to be the target gene of miR-20a.Compared with the normal group (24 h,48 h,72 h:0.23 ± 0.01,0.39 ± 0.01,0.56 ± 0.03) and the no-load group (24 h,48 h,72 h:0.25 ± 0.01,0.44 ± 0.05,0.59 ± 0.01),miR-20a did not change the cell proliferation at different time points(24 h,48 h,72 h:0.26 ± 0.01,0.41 ± 0.02,0.58 ± 0.02) (all P ＞ 0.05).Compared with the normal group (1.00 ± 0.05,1.24 ± 0.20,1.31 ± 0.09,0.89 ± 0.12) and the no-load group (0.76 ± 0.10,1.31 ± 0.13,1.50 ± 0.11,1.01 ± 0.11),miR-20a up-regulated the mRNA expressions of SP-A,SP-B,SP-C and SP-D (2.05 ± 0.17,2.14 ± 0.10,2.84 ± 0.09,1.66 ± 0.08),and the differences were significant (all P ＜ 0.05).Compared with the normal group (0.46 ± 0.01,0.27 ± 0.03,0.69 ± 0.01,0.43 ± 0.01) and no-load group (0.43 ± 0.01,0.21 ± 0.01,0.79 ± 0.02,0.44 ± 0.02),miR-20a also increased the protein expressions of SP-A,SP-B,SP-C and SP-D (0.55 ±0.01,0.47 ±0.05,0.96 ±0.02,0.59 ±0.03),the diffe-rences were statistically significant (all P ＜0.05).The expression of STAT3 in miR-20a group(0.37 ±0.05) was significantly lower than that in the normal group(0.60 ±0.04) and the no-load group (0.68 ±0.06),and the differences were statstically significant (all P ＜ 0.05) in A549.Conclusions STAT3 is a downstream target gene of miR-20a.miR-20a can promote pulmonary surfactant synthesis of alveolar epithelial cells A549 by inhibiting STAT3.
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Objective To study the roles of miR-20a in lipopolysaccharide induced inflammation of A549 cells and the possible mechanisms.Method The miR-20a mimic/inhibitor were transfected into A549 cells, and the cells were stimulated using lipopolysaccharide for 24 h.Interleukin-6 ( IL-6) and IL-8 were detected at mRNA level and protein level using real-time PCR and ELISA method , respectively.Protein expression of apoptosis signal regulating kinase 1 (ASK1)、P38、P-P38、JNK and P-JNK were detected using Western blot. Result Compared to mimic negative control group , the levels of mRNA and protein expression of IL-6 and IL-8 in the mimic group were all significantly decreased ( P<0.05).Compared to inhibitor negative control group , the levels of mRNA and protein expression of IL-6 and IL-8 in the inhibitor group were all significantly increased (P<0.05).The levels of ASK1, P-P38 and P-JNK protein in the mimic group were significantly lower than the mimic negative control group (P<0.05);the level of protein expression of ASK1, P-P38 and P-JNK in the inhibitor group were all higher than the inhibitor negative control group (P<0.05).Conclusion The regulation of ASK1 by miR-20a may play an important role in the inflammation process of acute respiratory distress syndrome .