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Objective:To investigate the effects of disialyllacto-N-tetraose (DSLNT) on low molecular weight metabolic profile of intestinal contents in neonatal rats with necrotizing enterocolitis (NEC), in an attempt to explore the protective mechanism of DLSNT on intestinal tract of neonates.Methods:Immediately after birth, SD rats were randomly divided into the control group, the NEC group and the NEC+ DSLNT group according to random number tale method.All rats were hand-fed by special formula milk.Rats in the NEC group and NEC+ DSLNT group were exposed to hypoxia (950 mL/L nitrogen, 10 min, thrice per day) and cold stress (4 ℃, 10 min, thrice per day) for continuous 3 days to establish rodent NEC model.Rats in the NEC+ DSLNT group were hand-fed with special formula containing 300 μmol/L DSLNT.All rats were sacrificed after 72 h, and intestinal contents were collected from ileum and colon, followed by untargeted metabolomic determination with the ultrahigh-performance liquid chromatography Q extractive mass spectrometry (UHPLC-QE-MS) method.The terminal ileum was examined by hematoxylin-eosin staining.The metabolome data were analyzed with multivariable analysis using SIMCA 14.1.The metabolites that met both variable importance in the projection (VIP) >1 in the orthogonal partial least squares analysis (OPLS-DA) model and P<0.05 in the t-test were screened as differential metabolites between groups. Results:DSLNT reduced the incidence of NEC and pathological scores of ileum tissue from neonatal rats with NEC [3.0(2.0, 3.0) scores vs.1.0(1.0, 2.0) scores, P<0.01], and also significantly suppressed inflammatory infiltration.OPLS-DA model based on the metabolome data determined by UHPLC-QE-MS could perform effective discrimination between the NEC group and the control group, as well as the NEC+ DSLNT group and the NEC group.There were 64 differential metabolites between the NEC group and the control group (VIP value>1 and P<0.05 for the OPLS-DA model). These metabolites included docosahexaenoic acid (+ 288.0%, P=0.028), xanthine (+ 372.1%, P=0.007), L-arginine (+ 233.1%, P=0.027), L-leucine (+ 232.7%, P=0.015), N-acetylneuraminic acid (-41.6%, P=0.014), and so forth.These metabolites were associated with 34 metabolic pathways.Among them, such 6 pathways as arginine biosynthesis, arginine and proline metabolism were the most disturbed pathways affected by NEC.There were 15 diffe-rential metabolites in between NEC+ DSLNT group and NEC group, which included D-mannose (-73.5%, P=0.032), xanthine (-63.4%, P=0.008), linoleic acid (+ 137.9%, P=0.047), nicotinamide adenine dinucleotide (+ 278.2%, P=0.005), and so forth.These metabolites were mapped to 7 metabolic pathways, among them, linoleic acid metabolism pathway was the most relevant differential pathway affected by DSLNT.There were 8 overlapped meta-bolites in both comparison strategies, and the variation trend of these overlapped metabolites in the NEC group was significantly reversed by DSLNT supplementation. Conclusions:DSLNT could significantly attenuate the NEC pathological damage caused by hypoxia/cold stress in neonatal rats.This protective effect is associated with the improvement of the metabolic profile of intestinal contents caused by NEC and the modulation of the linoleic acid metabolic pathway.The early preventive supplementation of DSLNT is of great significance in maintaining neonatal intestinal homeostasis and preventing the process of NEC.
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Objective To study the change and characterization of metabolic profile of intestinal contents of the neonatal rats with necrotizing enterocolitis (NEC) using metabolomics approach,in order to figure out potential biomarkers of NEC.Method Twenty rats with three-postnatal day-old fed with special formula were assigned to control group (n =8) and NEC group (n =12) randomly.Experimental NEC of rats in NEC group were induced by exposing to cold stimulation at 4 degrees Celsius for 10 minutes and to hypoxia at 95％ nitrogen for 10 minutes,three times a day for three consecutive days.All the rats were sacrificed after model preparation.Segments of the ileum of all the rats were collected for hematoxylin-eosin staining and subsequent pathological damage evaluation.The intestinal contents of the ileum and colon were collected by perfusion,followed by lyophilization and analyzed by UHPLC-QE-MS in order to conduct the non-target metabolomic determination.The information of the metabolites determined was calculated by multivariable analysis using SIMCA software.Result The pathological damage scores of NEC group were higher than those of the control group [(3.13 ± 0.83) vs.(0.25 ± 0.46),P ＜ 0.001].The results of orthogonal partial least squares discriminant analysis (OPLS-DA) model showed that in the ESI + mode,R2(x) =0.604,R2(y) =0.583,Q2 =0.960,while in the ESI-mode,the OPLS-DA model R2(x) =0.828,R2(y) =0.999,and Q2 =0.713,indicating that there is a significant difference in the intestinal content metabolic profile between the control group and the NEC group.Forty-eight differential metabolites related to NEC were identified.In ESI-mode,there were 22 differential metabolites,including L-isoisoleucine (+ 221％) and D-phenylalanine (+ 230％),L-histidine (+ 284％),xanthine (+ 207％),glutamyl leucine (+ 246％),allose (-70％),myristic acid (-57％) and pentadecanoic acid (-35％).What is more,in the ESI + mode,26 other differential metabolites were identified,including ornithine (+ 268％),D-leucine (+ 176％),L-iso Leucine (+ 213％),acetylcholine (+ 195％),nicotinamide adenine dinucleotide (+ 199％),citrulline (+ 158％),cytosine (-58％),xanthoic acid (-64％).These metabolites were reflected to 33 different metabolic pathways in KEGG databases.The pathway enrichment analysis and pathway topology analysis with MetaboAnalyst indicated that the arginine and proline metabolic pathways,histidine metabolic pathways,and glutathione metabolic pathways were the top altered pathways in the condition of NEC.Conclusion The metabolic profile of intestinal contents in NEC rats was significantly different from that in normal rats,which was characterized by amino acid accumulation,mainly involving the metabolic pathways of arginine,proline,histidine and glutathione.The detection of intestinal contents metabolic profile,especially amino acid metabolize group may be of great significance for the diagnosis of NEC,and improving intestinal microenvironment may be the key strategy for the prevention and treatment of NEC.
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Infancy is a critical period for the formation and development of intestinal flora, the intestinal flora mainly comes from the exchange between matrix and infants.Cesarean section, early exposure to antibiotics and formula feeding may disrupt the establishment of microbiota and adversely affect health.Human milk oligosaccharides (HMOs) is a group of unconjugated polysaccharides with diverse structures.It is rich in breast milk, and can regulate the composition of infant intestinal flora and have a beneficial impact on infant′s intestinal health.In order to understand the characteristics of the infant gut bacteria colonize, and the effect of HMOs on the intestinal flora, this review is as follows in this regard.
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Objective To explore the functional properties of the brain's resting state networks in neonates with hypoxic ischemic encephalopathy.Methods Use the methods of functional MRI probabilistic tractography and graph theory to compare the global and local functional properties of the brain's resting state network between 12 cases of children with mild HIE and 14 cases of children with moderate/severe HIE.Results In terms of global topological properties,both the mild and moderate/severe group showed small-world properties.The γ and λ in the mild group were 2.450± 1.642 and 1.542±0.564 and in the moderate/severe group were 2.331± 1.554 and 1.353±0.672,respectively,which were in line with the characteristics of small-world properties (γ＞ 1 and λ ≈ 1).As far as local topological properties were concerned,the distribution of hub regions in the functional networks had smaller number of nodes in the moderate/severe group (8 nodes) than the mild group (14 nodes).The comparison of nodal efficiencies showed that the moderate/severe group had significantly reduced nodal efficiency in the left insula opercula,left supramarginal gyrus,left and right temporal pole and right middle temporal gyrus(the nodal efficiencies of the moderate/severe group:0.4089±0.0865,0.3377±0.1223,0.3842±0.0898,0.3508±0.1295,0.3564±0.0843;the nodal efficiencies of the mild group:0.4801±0.0762,0.4465±0.0898,0.4655 ±0.0812,0.4640±0.0690,0.4271±0.0636,all P＜0.05).Conclusion The topological structure of resting state functional network in children with moderate and severe hypoxic-ischemic encephalopathy is obviously backward than that in children with mild hypoxic-ischemic encephalopathy,which may be related to abnormal language,movement and cognitive function in the late stage.
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Neonatal necrotizing enterocolitis (NEC) is the most common neonatal gastrointestinal emergency,and its pathogenesis is still not clear.In recent years,intestinal flora has been keenly concerned about its pivotal role in the progress of NEC pathology.At the technical level,the popularity and in-depth development of high throughput sequencing,fluorescence in situ hybridization and polymerase chain reaction-denaturing gradient gel electrophoresis are also favorable conditions for further study of the correlation between NEC and intestinal flora.Now,the relationship between the pathological development of NEC and intestinal flora was reviewed.
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Objective To discuss the prognostic evaluation of magnetic resonance diffusion tensor imaging (DTI) in term neonates with mild and severe asphyxia.Methods Eleven neonates with mild asphyxia,14 neonates with severe asphyxia and 10 control neonates were studied.All the neonates were examined by conventional magnetic resonance imaging(MRI) and DTI.Fraction anisotropy (FA) values,apparent diffusion coefficient (ADC) values,voxel numbers and fiber numbers were calculated in seven regions of interest(ROI) and compared among the 3 groups.The correlation between FA values and neonatal behavioral neurological assessment (NBNA) scores were analyzed.Results (1) FA values in the left and the right thalamus were 0.54 ± 0.08 and 0.56 ± 0.15 in control group,0.45 ±0.03 and 0.44 ± 0.10 in mild group,and 0.21 ± 0.11 and 0.25 ± 0.13,respectively in severe group.FA values in posterior limbs of internal capsule were 0.49 ± 0.09 and 0.48 ± 0.08 in control group,0.37 ± 0.08 and 0.38 ± 0.03 in mild group,and 0.20 ± 0.04,0.19 ± 0.13 in severe group;FA values in thalamus and posterior limbs of internal capsule had statistical differences among the 3 groups (F =9.12,9.11,8.18,8.55,all P ＜ 0.05).Voxel numbers in the left and the right superior longitudinal fasciculus were 1 094 ± 112 and 1 123 ± 113 in control group,986 ± 111 and 1 009 ± 144 in mild group,450 ± 116 and 671 ± 126 in severe group.Voxel numbers in anterior limbs of internal capsule were 947 ± 104 and 1 237 ± 184 in control group,854 ± 118 and 799 ± 114 in mild group,324 ± 110 and 311 ± 126 in severe group.Voxel numbers in posterior limbs of internal capsule were 2 047 ± 129 and 2 137 ± 238 in control group,1 843 ± 233 and 1 753 ± 247 in mild group,867 ± 118 and 999 ± 167 in severe group.Voxel numbers in superior longitudinal fasciculus,anterior and posterior limbs of internal capsule had statistical differences among the 3 groups (F =10.11,9.45,7.33,8.45,12.65,11.23,all P ＜ 0.05);Fiber numbers in the left and the right cingulate gyrus were 245 ±72 and 405 ±94 in control group,225 ±52 and 365 ± 114 in mild group,145 ±62 and 185 ±84 in severe group.Fiber numbers in inferior front-occipital fasciculus were 56 ± 19 and 212 ± 33 in control group,49 ±22 and 197 ± 33 in mild group,33 ± 12 and 156 ± 39 in severe group.Fiber numbers in posterior limbs of internal capsule were 284 ± 112 and 988 ± 233 in control group,234 ± 67 and 678 ± 234 in mild group,114 ± 67 and 188 ± 84 in severe group.Fiber numbers in cingulate gyrus,inferior front-occipital fasciculus and posterior limbs of internal capsule had statistical differences among the 3 groups (F =3.11,3.45,9.88,9.12,7.45,8.88,all P ＜ 0.05).(2) The correlation between FA value and NBNA scores was analyzed:the correlation coefficient in posterior limbs of internal capsule was 0.666 which was higher than those of the other areas.The area under the ROC curve of FA values in anterior limbs of internal capsule was 0.816 (P =0.005),used NBNA scores ≥ 35 as a good outcome.FA values (≥ 0.375) in the posterior limbs of the internal capsules predicted a good outcome and the corresponding sensitivity and specificity was 73.1％ and 77.8％,respectively.Conclusion In some ROIs,FA values,voxel numbers,and fiber numbers can quantitatively reflect the degree of white matter injury in neonates with asphyxia.Furthermore,the FA values in the posterior limbs of the internal capsules are closely correlated with NBNA scores,so it has more important clinical significance,and can accurately and objectively assess the prognosis in neonates with asphyxia.
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Necrotizing enterocolitis (NEC) is one of the common intestinal critical disease in newborns especially in premature infants.However,the etiology and pathogenesis of NEC are not fully clear.There were a lot of research which had shown that breast-feeding could significantly reduce the incidence of NEC.The composition in human milk,such as human milk oligosaccharides,growth factors,glutamine,lactoferrin,has effects on anti-infection,maintaining intestinal mucosal integrity and immune regulation,and the proportion of nutrients,and osmotic pressure in human milk is especially suitable for the needs of gastrointestinal physiological of premature infants,which can effectively prevent the happening of the NEC.Now,the research progress on components in human milk and their mechanism for preventing necrotizing enterocolitis was reviewed.
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Objective To investigate the characteristics of cognitive development of newborn's cerebral hemispheres with event related potential (ERP). Methods Forty-five healthy full-term newborns and 11 infants of 29-60 days in the Department of Newborns, Changzhou Children's Hospital between December 2013 and October 2015 were randomly included in this study. They fulfilled all inclusion/exclusion criteria. An auditory oddball paradigm was used with frequently occurring standand tones at 1 000 Hz with 90% probability and infrequent deviant tones at 2 000 Hz with 10% probability. Waveforms were recorded at Fz, Cz, F3, F4, C3 and C4 scalp locations. And a comparative study of area and latency of N2 was done at F3, F4, C3 and C4 scalp locations in the two hemispheres. Results N2 wave of 1-28 days neonatal at C3, C4 scalp locations: area (ms·μV) was 5 193.68±2 137.45 vs 4 324.51±1 334.73, with statistically significant difference (t=2.288, P=0.025);latency (ms) was 764.27±216.60 vs 878.29±294.06, also with statistically significant difference (t=2.185, P=0.031). N2 wave of 1-28 days neonatal at F3,F4 scalp locations: area (ms·μV) was 5 739.44±2 271.03 vs 5 614.87±2 280.83, without statistically significant difference;latency (ms) was 876.20±316.02 vs 860.45±289.04, also without statistically significant difference. N2 wave of 29-60 days infants at F3, F4 scalp locations: area (ms·μV) was 7 681.35±1 723.49 vs 5 993.55±735.63, with statistically significant difference (t=2.848, P=0.011);latency (ms) was 571.60±1.2248 vs 733.40±232.73, without statistically significant difference though F3 was shorter than F4. Conclusions The newborn's auditory perception cognition in the left side of central lobe is superior to the right side. With the growth of the newborn, the left side of the frontal lobe is also superior to that of the right side. Dominant hemisphere has appeared in the neonatal period. Proper stimulation can promote the newborn's cognitive development.
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Objective To analyze the biological activity of bioactive peptides in human breast milk and to find the polypeptides so as to investigate the preventive and therapeutic effects of breast milk-derived bioactive peptides on neonatal necrotizing enterocolitis(NEC).Methods Six mothers who gave birth to preterm neonates were enrolled in this study and 5 mL of their breast milk secreted within 2-5 postnatal days were collected for 6 times and blended subsequently.Bioactive peptides from maternal milk of the preterm infants were separated by ultrafiltration and analyzed by using tandem mass spectrometry.Polypeptides possibly with biological function were screened out by using bioinformatics software and the protein function cluster online analysis software was used to predict the polypeptides associated with infection according to the biological function of their precursor proteins.The ATCC25922,an Escherichia coli strain commonly associated with infection in NEC and drug solution (sulbactarr/cefoperazone) were used to conduct the drug susceptibility testing and bactericidal kinetics testing,so as to verify the antibacterial effects of bioactive peptides in the breast milk.Results Four thousand three hundred and eleven peptides contained in breast milk were identified successfully,of which 1 370 were non-differential peptides,and 188 peptides possibly with biological activity and 11 peptides were associated with infection.The peptide compound in the breast milk had antimicrobial activity and bactericidal power against Escherichia coli.Conclusions The active peptide compounds in the breast milk have antimicrobial activity,which play an important role in the prevention of NEC.Finding the true antimicrobial peptides with in vivo and in vitro biological activity by using antimicrobial spectrum test is expected.
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Objective To discuss the prognostic evaluation of magnetic resonance diffusion tensor imaging (DTI) in term neonates with mild and severe asphyxia.Methods Eleven neonates with mild asphyxia,14 neonates with severe asphyxia and 10 control neonates were studied.All the neonates were examined by conventional magnetic resonance imaging(MRI) and DTI.Fraction anisotropy (FA) values,apparent diffusion coefficient (ADC) values,voxel numbers and fiber numbers were calculated in seven regions of interest(ROI) and compared among the 3 groups.The correlation between FA values and neonatal behavioral neurological assessment (NBNA) scores were analyzed.Results (1) FA values in the left and the right thalamus were 0.54 ± 0.08 and 0.56 ± 0.15 in control group,0.45 ±0.03 and 0.44 ± 0.10 in mild group,and 0.21 ± 0.11 and 0.25 ± 0.13,respectively in severe group.FA values in posterior limbs of internal capsule were 0.49 ± 0.09 and 0.48 ± 0.08 in control group,0.37 ± 0.08 and 0.38 ± 0.03 in mild group,and 0.20 ± 0.04,0.19 ± 0.13 in severe group;FA values in thalamus and posterior limbs of internal capsule had statistical differences among the 3 groups (F =9.12,9.11,8.18,8.55,all P ＜ 0.05).Voxel numbers in the left and the right superior longitudinal fasciculus were 1 094 ± 112 and 1 123 ± 113 in control group,986 ± 111 and 1 009 ± 144 in mild group,450 ± 116 and 671 ± 126 in severe group.Voxel numbers in anterior limbs of internal capsule were 947 ± 104 and 1 237 ± 184 in control group,854 ± 118 and 799 ± 114 in mild group,324 ± 110 and 311 ± 126 in severe group.Voxel numbers in posterior limbs of internal capsule were 2 047 ± 129 and 2 137 ± 238 in control group,1 843 ± 233 and 1 753 ± 247 in mild group,867 ± 118 and 999 ± 167 in severe group.Voxel numbers in superior longitudinal fasciculus,anterior and posterior limbs of internal capsule had statistical differences among the 3 groups (F =10.11,9.45,7.33,8.45,12.65,11.23,all P ＜ 0.05);Fiber numbers in the left and the right cingulate gyrus were 245 ±72 and 405 ±94 in control group,225 ±52 and 365 ± 114 in mild group,145 ±62 and 185 ±84 in severe group.Fiber numbers in inferior front-occipital fasciculus were 56 ± 19 and 212 ± 33 in control group,49 ±22 and 197 ± 33 in mild group,33 ± 12 and 156 ± 39 in severe group.Fiber numbers in posterior limbs of internal capsule were 284 ± 112 and 988 ± 233 in control group,234 ± 67 and 678 ± 234 in mild group,114 ± 67 and 188 ± 84 in severe group.Fiber numbers in cingulate gyrus,inferior front-occipital fasciculus and posterior limbs of internal capsule had statistical differences among the 3 groups (F =3.11,3.45,9.88,9.12,7.45,8.88,all P ＜ 0.05).(2) The correlation between FA value and NBNA scores was analyzed:the correlation coefficient in posterior limbs of internal capsule was 0.666 which was higher than those of the other areas.The area under the ROC curve of FA values in anterior limbs of internal capsule was 0.816 (P =0.005),used NBNA scores ≥ 35 as a good outcome.FA values (≥ 0.375) in the posterior limbs of the internal capsules predicted a good outcome and the corresponding sensitivity and specificity was 73.1％ and 77.8％,respectively.Conclusion In some ROIs,FA values,voxel numbers,and fiber numbers can quantitatively reflect the degree of white matter injury in neonates with asphyxia.Furthermore,the FA values in the posterior limbs of the internal capsules are closely correlated with NBNA scores,so it has more important clinical significance,and can accurately and objectively assess the prognosis in neonates with asphyxia.
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Objective:To investigate the changes of gene expression profile in lipopolysaccharide (LPS)-stimulated primary Kupffer cells ( KCs ) . Methods: Rat KCs were isolated and purified by means of in situ perfusion and density gradient centrifugation. After being identified by ink phagocytosis and ED2 staining test,KCs were stimulated with LPS. Gene expression profile were studied using gene microarrays,and the most significant upregulated gene was verified using real-time PCR. Results:27 genes were upregulated including Ces1f, Slc17a3, Slc21a4, Hsd17b2, Sorbs2, Ccdc116, Mgam, Myo5b, Etl4, Fabp1, Kif4b, Fosl1, Cyp4a1, Penk, Tmem221,Rpl5,Nr2f1,Hoxb1,Gpr165,Fam90a13p,Kpna6,Irak1bp1,Kcnh1 and 4 unnamed genes and 4 downregulated including Oc90,Tagln,Arxes2 and Olr830 in LPS-stimulated KCs. Among the upregulated genes, Ces1f was the most significant upregulatory gene. Real-time PCR confirmed that the levels of Ces1f were 23. 88 times higher in LPS-stimulated than control cells. Conclusion:There is a significant difference between LPS-stimulated and normal control cells in gene expression profile by microarray analysis,and Ces1f is the most significantly upregulated gene.
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Objective:To investigate the effects of IRF 3 shRNA adenovirus on dynamic changes of early cytokines in LPS-stimulated primary Kupffer cells ( KCs ).Methods: Rat KCs were isolated and purified by means of in situ perfusion.After being infected with adenovirus carrying IRF 3 shRNA for 48 h, KCs were stimulated with LPS.Cell culture supernatants were collected respectively at 0,2,4 and 6 h after LPS stimulation as well as cells at 6 h.Supernatant cytokine secretion levels were detected by enzyme-linked immunosorbent assay ( ELISA).Intracellular gene expressions were tested by RT-PCR and Westeron blot.Results:IRF3 mRNA and protein were induced by LPS ,but suppressed by IRF 3 shRNA adenovirus in LPS-stimulated or non-stimulated KCs.IFN-βsecretions rose in the very early stage ( at 2 h) ,reached the peak at 4 h,and began to reduce but still remained high levels at 6 h after LPS stimulation in KCs.Interference adenovirus pretreatment suppressed IFN-βsecretions ( especially the secretion peak ) at each time point after LPS stimulation.IFN-βsecretions reached normal levels at 6 h after the stimulation in adenovirus-pretreated cells;TNF-αse-cretions rapidly increased in the very early stage and reached the peak at 2 h,then began to decrease gradually ,but remained high levels at 6 h after LPS stimulation in KCs.Interference adenovirus pretreatment inhibited LPS-induced TNF-αsecretions, especially the secretion peak;IL-1βsecretions did not increase untill 4 h, but reached a higher level at 6 h after LPS stimulation.Interference adenovirus suppressed IL-1βsecretions in the early stage of LPS stimulation;IL-10 secretions began to rise in the very early stage ,and gradually increased over time after LPS stimulation in KCs.Pretreatment of adenovirus with IRF 3 shRNA promoted upregulations of IL-10 secretions at each time point of the early of LPS stimulation.Conclusion:IRF3 gene expression can be silenced by IRF 3 shRNA ad-enovirus.IRF3 can promote its downstream signaling molecule IFN-βand pro-inflammatory cytokines including TNF-αand IL-1β,and block anti-inflammation cytokine IL-10 secretions in LPS-stimulated primary KCs.Therefore,IRF3 may play a central role in immune inflammatory injury of liver tissues.
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Cognitive function is the advanced features of brain and neonatal cognitive function has its own development characteristics. Quantitative electroencephalogram can reflect the state of brain function directly and objectively and can be used in cognitive research. Event-related potential,induced by stimulation,is the neu-roelectrophysiologic of brain cognitive processing and time-locked to cognitive events. In recent years,research-ers have made great progress in exploring the neonatal cognitive function with the help of quantitative electroen-cephalogram and event-related potential. This review summarizes the progress of these two technologies in neo-natal brain cognitive function.
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Objective To investigate the characteristics of cognitive development in different aged neonates.Methods Sixty-two newborns were randomly selected from relatively normal full-term babies in Changzhou Children's Hospital from December 2013 to September 2015.Electroencephalogram (EEG) and event-related potentials (ERP) were recorded with the auditory Oddball paradigm.Cognitive EEG delta power and the N2 wave area of different ages (1-10,11-20 and 21-28 days) were compared.Paired t test,analysis of variance and the LSD test were used for statistical analysis.Results (1) Delta power in the resting and cognition state:neonatal cognitive delta power in the 11-20 and 21-28 days old groups was (268.22± 132.09) and (236.01±97.40) μ V2,respectively,significantly higher than the resting delta power of the same groups [(175.80 ± 80.80) and (178.78 ± 104.74) μ V2,t=2.539 and 2.845,P=0.020 and 0.010,respectively].(2) Cognitive delta power in different aged neonates:cognitive delta power in the 11-20 and 21-28 days old groups was (268.22± 132.09) and (236.01 ±97.40) μ V2,respectively,higher than that of the 1-10 days old group [(116.70± 56.70) μV2],with statistically significant difference (LSD test,both P＜0.05).(3) Neonatal ERP:ERP of the 1-10 days old group presented with multiple peaks of a flat composite wave,in the 11-20 and 21-28 days old groups,the N2 wave showed a regular and rising trend,gradually to a single wave,and became gradually mature.The N2 wave area in the 11-20 and 21-28 days old groups was (6 435.08±2 212.34) and (6 536.75± 1 969.86) ms · μ V,respectively,which was larger than that in the 1-10 days old group [(4 230.04± 1 550.55) ms · μ V] (LSD test,both P＜0.05).Conclusions Neonatal cognitive development is enhanced with age and there may be a period of more rapid cognitive development,especially at 11-20 days of age.
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Objective To investigate the impacts of adenovirus on the transfection efficiency and proliferative activity of primary Kupffer cells (KCs). Methods Rat liver KCs were separated and purified by density gradient centrifugation , and was then transfected with adenovirus carrying green fluorescence protein (GFP) gene at different multiplicity of infection (MOI). After 24 h, the transfection efficiency was evaluated by fluorescence microscope and flow cytometry. The proliferative activity of KCs was assessed by colorimetric method. Results The positive percentages of GFP staining cells were statistically different among different doses of adenovirus (MOI 0, 100, 300, 500, 700 and 900) under fluorescence microscopy or by flow cytometry (P 0.05 for all comparisons) by CCK8 assay. Conclusions KCs can effectively be transfected by GFP adenovirus; and with an increase in virus MOI, the transfection efficiency rises gradually. A higher dose of adenovirus may have a negative effect on cell proliferative.
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Objective To explore Chinese character semantic processing feature and the relationship with behavioral problems in early childhood with ADHD.Methods Twenty ADHD children and twenty normal children (7-8 years old) performed a Chinese character semantic judgment task through event related potential (ERP),meanwhile Conners parent rating scale (PSQ) was completed by parents to analyze behavior problems in children.The relationship between behavioral problems and ERPs was analyzed.Results (1) ADHD group showed smaller amplitudes of P200 compared to the normal group (related (6.6±4.8) μV vs (11.6±6.4) μV;unrelated (6.7±3.7) μN vs (10.8±5.0) μV),and longer latency (related(240±30) ms vs (198±17)ms;the unrelated (232±26)ms vs (194±16)ms;psudoword (233±26)ms vs (197±15)ms),all P＜0.05).The amplitudes of N400 wave only showed a decreasing trend in ADHD group(P＞0.05),while the latency of N400 was significantly longer than that in normal children in three conditions(P＜0.05).(2) Four subscale scores of CPSQ(conduct problem,learning problem,hyperactivity-impulsion and hyperactivity index) were significantly higher in ADHD children compared with normal group.(3)The amplitudes of P200 in related condition had negative correlation with the scores of CPSQ subscale (conduct problem,psychoso matic disorder,hyperactivity-impulsion and hyperactivity index) (r=-0.51,-0.47,-0.45,-0.46,all P＜0.05.The latency of P200 in related condition had positive correlation with psychosomatic disorder scores (r =0.54,P=0.01).Conclusion Early semantic recognition disorder exists in 7～ 8 years old ADHD children.P200 can be used as an objective index to evaluate early childhood ADHD's language cognition,and P200 is related to behavior problems (such as conduct problem,impulsions).
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Objective To discuss the value of magnetic resonance (MR) diffusion tensor imaging (DTI) technology in the diagnosis of the apparent diffusion coefficient (ADC) and the fraction anisotropy (FA) of neonates with hypoxic-ischemic encephalopathy (HIE) in the subacute stage.Methods Ten cases of normal full-term neonates and 34 term neonates diagnosed with HIE in the subacute stage were investigated with conventional MR and DTI.Of the 34 cases, 15 cases were mild HIE, 12 cases were moderate HIE,7 cases were severe HIE.ADC/FA values in different parts of the brain were analyzed.And the receiver operating characteristic (ROC) curve was used,the sensitivity and specificity of the FA in specific parts were also analyzed.Results (1) On convention MRI : according to the MRl scoring system,score of mild group was(2.6 ±-1.7) points, moderate group was (3.0 ± 1.5) points, severe group was (5.8 ± 1.3) points.There was significant difference between mild group and severe group, moderate and severe group (P ＜ 0.05).(2) The ADC and FA values in each group on DTI : ADC values in all of the regions of interest (ROIs) had no statistic difference between the HIE group and the controls(all P ＞ 0.05).FA values in posterior limb of internal capsule (PLIC) ,splenium had statistic difference between the HIE group and the controls.FA values in mild group were (0.317-0.018,0.339 ±0.025) ,moderate group (0.295 ±0.025,0.324 ±0.026) and severe group (0.252 ±0.039,0.253-0.036) and control group (0.455 ± 0.047,0.372 ± 0.032).(3) ROC analysis of the neonates diagnosed with HIE:the area under the ROC curve of FA values in PLIC, Splenium had statistic difference between mild group and moderate group (P ＜ 0.05).The area under the ROC curve in each group was 0.744 (P =0.032) ,0.658 (P =0.164).ROC curve was used to evaluate the diagnostic accuracy of FA values in PLIC between HIE group and control group,when FA value was lower than 0.337,0.803 was the largest area under the ROC curve, and there was significant difference between them(P ＜ 0.01).Conclusions FA values can reflect the brain white matter WM injury of HIE neonates quantitatively,and its diagnostic veracity is better than ADC value and conventional MR during the subacute phase of HIE.The diagnostic veracity of FA values in PLIC is higher than other parts.When the FA value is lower than 0.337, there is good sensitivity and specificity in diagnosis of HIE, so DTI technology can provide accurate and objective basis for clinical treatment with good sensitivity and specificity about the diagnosis of HIE in early stage.
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Objective To study the characteristics of cognitive development of newborns with auditory event related potentials (aERP).Methods Thirty full-term neonates who were admitted to Changzhou Children's Hospital and met the inclusion criteria, those with neonatal neural behavioral score over 37 and both ears passed the hearing screening, and exclusion criteria, those who suffered from perinatal brain injury, were sclccted randomly.Thcy wcre divided into three groups according to the days of age (group 1 : 1-10 days;group 2:11-20 days;group 3:21-28 days).aERP was recorded with auditory Oddball paradigm.The data were collected and analyzed by Brain Electrical Source Analysis software.Relationship between N2latencies/areas among the three groups and their age was researched with Pearson correlation analysis.Results During the neonatal period, there was a significant negative correlation between N2 latency and neonatal age in days (r=-0.609, P ＜ 0.05), while significant positive correlation was shown between N2 area and the age (r=0.689, P ＜ 0.05).In Fz and Cz leads, the total average waveform diagram of the three groups showed spacious flat, less smoothed curve, and mostly jagged pattern.N2 area in group 1 showed a complex plurality of peaks, and regular, soaring waves, which changed into single wave thereafter, in group 2 and 3.The N2 latencies tended to shorten gradually.Conclusions Neonatal cognitive competence, including concentration and discrimination ability, would be improved with their growth.
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Objective To investigate the effects of interferon regulator factor 3 (IRF3) shRNA on the expression of TLR4 downstream signal molecules including IRF3-IFN-β, NF-κB/p38 MAPK-TNF-α/IL-1βand IL-10 in lipopolysaccharide (LPS)-stimulated Kupffer cells (KCs). Methods KCs were isolated from rats by in situ perfusion. The adenovirus strains carrying IRF3 shRNA were used for the transfection of purified KCs. The isolated KCs were randomly divided into four groups including adenovirus(-) LPS(-) treatment group, adenovirus(-) LPS(+) treatment group, adenovirus(+) LPS(-) treatment group and ad-enovirus(+) LPS(+) treatment group. The levels of cytokines in the supernatants of KC culture were detec-ted by enzyme-linked immunosorbent assay ( ELISA ) . Real-time PCR and Western blot assay were per-formed to analyze the expression of related cytokines at mRNA and protein levels, respectively. Results The expression of IRF3 at mRNA and protein levels in primary cultured KCs were induced by LPS. The cel-lular constitutive expression of IRF3 at mRNA level and the LPS-induced expression of IRF3 were signifi-cantly inhibited after transfection of KCs with adenovirus strains carrying IRF3 shRNA. However, the nucle-ar constitutive expression of IRF3 protein was not affected by IRF3 shRNA. The expression of IFN-βat mR-NA and protein levels in KCs were induced by LPS, but were suppressed by the interference with IRF3 shR-NA. No significant changes of the cellular constitutive expression of IFN-βat mRNA and protein levels were observed in IRF3 shRNA-treated KCs. Enhanced expression of proinflammatory cytokines including TNF-αand IL-1β at mRNA and protein levels were detected in LPS-stimulated KCs. Transfection of KCs with ade-novirus strains carrying IRF3 shRNA inhibited the LPS-induced secretion of TNF-α and IL-1β, but neither LPS-induced expression of TNF-α and IL-1β at mRNA level nor cellular constitutive expression of TNF-αand IL-1βat mRNA and protein levels were affected by IRF3 shRNA. The LPS-induced expression of IL-10 at mRNA and protein levels were enhanced in IRF3 shRNA-treated KCs. However, the cellular constitutive expression of IL-10 at mRNA and protein levels were not affected by the adenovirus. The levels of phosphor-NF-κB p65 subunit and phosphor-p38 MAPK protein in the nuclei of KCs were increased upon the stimula-tion with LPS. Treatment of KCs with IRF3 shRNA showed no significant effects on nuclear phosphor-NF-κB p65 subunit and phosphor-p38 MAPK. Conclusion Transfection of LPS-stimulated primary KCs with ade-novirus strains carrying IRF3 shRNA could effectively inhibit the expression of IRF3 and the transduction of downstream signals. IRF3 enhanced the secretion of TNF-αand IL-1β, but inhibited the expression of IL-10 in LPS-treated KCs. The LPS-induced activation of NF-κB and p38 MAPK in KCs were not affected by IRF3 signal.
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Objecitve:To investigate effects of urotensin Ⅱ( UⅡ)/UT system on innate immune inflammatory signal pathway TLR4-IRF3 in the lipopolysaccharide (LPS)-stimulated Kupffer cells (KCs).Methods: Rat KCs were isolated and cultured.Pro-in-flammatory cytokines including IL-6,IFN-βand IFN-γwere assayed by ELISA in culture supernatant of KCs.Cell surface TLR4 were tested with flow cytometry technique.Expression of IRF3 were tested with real-time PCR and Western blot.Results: Significant increases were showed in IL-6, IFN-βand IFN-γsecretion, TLR4-expressed positive rates and IRF3 mRNA levels in KCs after stimulated by LPS,but were inhibited via urantide pretreatment.In addition,LPS induced upregulation of nuclear IRF3 protein and downregulation of cytoplasm IRF3 protein in KCs,which were blocked by urantide pretreatment.Conclusion:UⅡ/UT system mediates immune inflammatory response in part through activating TLR 4-IRF3 pathway in LPS-stimulated KCs.