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.

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.

Newborn brain damage is the main cause of new-borns' death and disabilities.Current research difficulties lie in analyzing characteristics of cerebral injuries,making objective prognosis and early intervention,as well as analysis of therapeutic effects after recovery.Since subjects are not requested to complete complex tasks while doing resting state functional MRI (rs-fMRI) tests,rs-fMRI is reckoned to be suitable for neonatal brain function research.So far,most rs-fMRI reports regarding cerebral injury are for adults,with only a few have been done on neonates.Foreign research are mainly focused on new borns' brain development.If relevant rs-fMRI research can be done on newborn brain damage,it would be helpful to accurately evaluate structure and function of patients' brain tissue damage.Further research can provide more valuable information in clinics.

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.

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.

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.

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). The cell proliferative activity had significant differences among MOI 300, 500, 700 and 900(P < 0.05 for all comparisons), but had no differences among MOI 0, 100 and 300 (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.

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.

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.

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.