Immunomodulatory effect and molecular mechanism of deoxyribonucleic acid receptor toll-like receptor 9 signaling pathway on newborn babies with acute lung injury.

This work was to demonstrate the immunomodulatory effect of toll-like receptor 9 (TLR9) signaling on newborn babies with acute lung injury (ALI) and the mechanism of TLR9 in vivo, so as to provide experimental basis for clinical treatment of newborn babies with ALI. Firstly, the expression of TLR9 in peripheral blood mononuclear cell (PBMC) was compared among ALI and healthy newborn babies. Then, PBMCs of newborn babies with ALI were extracted and divided into three groups. They were added with non-methylated cytosine purine-guanine dinucleotide sequence oligodeoxyribonucleotide (CpG ODN), ODN without non-methylated CpG, and blank nutrient solution, respectively, so as to determine the proliferation changes of PBMC. The immunohistochemistry (IHC) method was applied to detect the protein expression of TLR9-myeloid differentiation factor 88 (MyD88) signaling in lung tissue, and the number of T cell subsets (CD3+, CD4+, and CD8+) was detected by flow cytometry. Besides, enzyme-linked immunosorbent assay (ELISA) was employed to determine the concentration of interferon-α (INF-α) and INF-γ. The results revealed a neglectable difference in TLR9 expression in PBMCs among ALI and healthy newborn babies (P>0.05). Additionally, the proliferation number of PBMC cultured in CpG ODN group was greatly superior to the number of ODN and blank groups (P<0.05), and the INF-α and INF-γ of CpG ODN group increased obviously versus those of blank and ODN groups (P<0.05). In conclusion, TLR9 in PBMCs was present in both ALI and healthy newborn babies. CpG ODN could specifically recognize the TLR9 signaling, so as to activate the immune function of T lymphocyte subsets in newborn babies with ALI and promote the release of inflammatory factors from the neonatal patient's immune cells, thereby mediating the immune response of neonatal patients.

Prognostic Value of Ionized Calcium Levels in Neonatal Sepsis.

BACKGROUND: Despite recent advances in the treatment of neonatal infection, mortality rates and comorbidities associated with neonatal sepsis remain high. Hypocalcemia has been reported in critically ill patients, especially in as-sociation with sepsis. However, the importance of hypo-calcemia in neonatal sepsis has not been explored in detail. OBJECTIVES: The purpose of this study was to evaluate the prognostic value of hypocalcemia in neonatal sepsis patients and to identify the risk factors associated with sepsis-related mortality. METHODS: This retrospective study examined perinatal data from patients in a level IV neonatal in-tensive care unit between January 2010 and June 2016. Univariate analysis was performed to understand the differences in clinical and laboratory characteristics between patients with and without neonatal sepsis. Neonates with sepsis were further stratified as having ionized hypocalcemia (if serum ionized calcium [iCa] <1.0 mmol/L) or not. Uni- and multivariate logistic regression analyses were utilized to evaluate the predictive potential of iCa for identifying sepsis-related mortality. RESULTS: A total of 472 neonates were enrolled in this study, including 169 neonates diagnosed with culture-proven sepsis and 303 neonates without infection (control group). The comparison of neonates with and without sepsis highlighted significant differences in levels of iCa (0.97 ± 0.26 vs. 1.12 ± 0.25 mmol/L), magnesium (0.75 ± 0.22 vs. 0.89 ± 0.12 mmol/L), and phosphate (2.26 ± 1.08 vs. 1.65 ± 0.85 mmol/L; all p < 0.001). When neonates with sepsis were stratified into 2 subgroups based on serum iCa, neonates with hypocalcemia showed higher rates of organ dysfunction than those with normal iCa, as well as higher rates of cardiovascular system dysfunction (37.35 vs. 17.44%), renal dysfunction (34.94 vs. 30.95%), disseminated intravascular coagulation (26.51 vs. 11.63%), and seizure (16.04 vs. 5.8%; all p < 0.05). Among all neonates who had sepsis, the mortality rate was 13.61%, and this rate was higher among neonates with hypocalcemia than among those with normal iCa (20.48 vs. 6.98%, p < 0.05). Uni- and multivariate analyses showed that acidosis, hypoalbuminemia, hypocalcemia, and hyperphosphatemia were independent prognostic markers of sepsis-related mortality. In receiver-operating characteristic curve analysis, the areas under the curve were 0.70 (95% CI 0.624-0.768; p = 0.0004), 0.74 (95% CI 0.671-0.808; p < 0.0001), 0.73 (95% CI 0.653-0.792; p = 0.0002), and 0.67 (95% CI 0.59-0.737; p = 0.0154) for serum albumin, iCa, phosphate, and acidosis, respectively. Based on these findings, we developed a nomogram to predict sepsis-related mortality. CONCLUSIONS: Hypocalcemia is common in neonates with sepsis and is significantly associated with organ dysfunction and sepsis-related mortality.

Surfactant protein b expression in bronchoalveolar lavage fluid of full-term neonates with respiratory distress syndrome.

The aim was to investigate the surfactant protein B (SP-B) expression in the bronchoalveolar lavage fluid (BALF) of full-term neonates with respiratory distress syndrome (RDS). The enzyme-linked immunosorbent assay was performed to assess SP-B expression in BALF of 60 full-term neonates with RDS and 23 healthy neonates and correlation of SP-B level with RDS classification according to chest x-ray findings and PaO2/FiO2 before mechanical ventilation in neonates with RDS. The SP-B level was significantly lower in the RDS group (17.63 +/- 6.80 ng/mL) than in healthy neonates (103.95 +/- 6.38 ng/mL) (P < 0.001). The SP-B level correlated positively with PaO2/ FiO2 before mechanical ventilation (r = 0.838, P < 0.001). Moreover, the lower the SP-B level, the more severe was the RDS as determined by chest x-ray (P < 0.001). In conclusion, full-term neonates with RDS had reduced SP-B in BALF, which was related to the severity of RDS, suggesting that SP-B supplement may be an effective strategy in the treatment of RDS in full-term neonates.

Value of amplitude-integrated electroencephalograph in early diagnosis and prognosis prediction of neonatal hypoxic-ischemic encephalopathy.

UNLABELLED: To investigate value of amplitude-integrated electroencephalograph (aEEG) in early diagnosis and prediction of long-term prognosis of neonatal hypoxic-ischemic encephalopathy (HIE), 120 HIE Children were randomly assigned into aEEG group and control group (n = 60 per group). Children in each group were sub-divided into mild, moderate and severe HIE groups (n = 20 per group). 1, 3, 14 and 28 days after birth, aEEG was performed in aEEG group; 3, 14 and 28 days after birth, neonatal behavioral neurological assessment (NBNA) was done in both groups. Children who discharged were followed up at adjusted gestational age of 12 months with Denver Developmental Screening Test (DDST) and prognosis evaluation. RESULTS: aEEG manifestation was positively related to clinical severityb of HIE (r = 0.843, P < 0.01). On day 3 and 14, NBNA score was comparable between two groups (P > 0.05), but significant difference in NBNA score was noted on day 28 (P < 0.05). On day 3, 14 and 28, aEEG manifestation was positively associated with prognosis at adjusted gestational age of 12 months (r = 0.832, 0.857, 0.778, 0.743, P < 0.01). In aEEG group, disability rate was 13.8%, which was significantly lower than that in control group (23.2%); cure rate in aEEG group (60%) was significantly higher than that in control group (40%). Moreover, long-term prognosis was also dramatically different between aEEG group and control group (χ(2) = 4.107, P < 0.05). CONCLUSION: aEEG manifestation is significantly associated with clinical severity of HIE and may be helpful for early diagnosis of HIE. aEEG may be used to predict long term prognosis of HIE children.

Development of neural stem cells at different sites of fetus brain of different gestational age.

OBJECTIVE: This study aimed to investigate the development of neural stem cells (NSCs) in fetal brain, which may provide experimental evidence for the clinical treatment of brain injury in children. METHODS: A total of 60 fetuses were collected after labor induction and divided into 6 groups according to the gestational age (16 w, 20 w, 24 w, 28 w, 32 w and 36 w; n=10 per group). The hippocampus, striatum, subventricular zone, frontal lobe, temporal lobe, occipital lobe and parietal lobe were harvested. In situ hybridization, immunohistochemistry and light microscopy were done to determine the morphology and quantity of NSCs. RESULTS: NSCs were identified in the brain of fetuses with different gestational age. NSCs were round, oval, spindle-shaped, starlike, triangular or polygonal. NSC colony was also observed with symmetrical or asymmetrical division. Single NSC, group-like NSCs and cluster-like NSCs were found in the different sites of fetal brain, and NCSs interacted with each other via synapses. However, the distribution, morphology, growth and quantity of NSCs were different in the brain of fetuses with different gestational age. The number of NSCs reduced with the increase in gestational age, but they were always observed. CONCLUSION: The morphology of NSCs in fetal brain is variable and they are widely distributed in the hippocampus, subventricular zone, striatum and cortex. The number of NSCs reduced with the increase of gestational age.

Role of mouse nerve growth factor in neural recovery following hypoxic-ischemic brain damage.

OBJECTIVE: To investigate the role of mouse nerve growth factor (mNGF) in neutral repair following hypoxic-ischemic brain damage (HIBD) in a neonatal rat model. METHODS: A total of 120 neonatal rats aged 7 days were randomly divided into control group, HIBD group and mNGF group (n=40 /group). Immediately after HIBD, mNGF was intramuscularly injected into rats in the mNGF group. 7, 14, 21 and 28 days after injection, immunohistochemistry and TUNEL staining were performed. RESULTS: In the HIBD group, Nestin expression was mainly found in the CA1, CA2 and CA3 regions of hippocampus. In the mNGF group, Nestin was mainly noted in the DG and CA3 regions. 7, 14 and 28 days after treatment, significant difference was found in the Nestin expression among three groups, but not observed 21 days after treatment. 21 and 28 days after treatment, the number of apoptotic neurons was markedly reduced when compared with that 7 and 14 days after treatment. CONCLUSION: mNGF can improve the neurogenesis and inhibit neural apoptosis in the hippocampus following HIBD in a neonatal rat model.

Effect of mouse nerve growth factor on the expression of glial fibrillary acidic protein in hippocampus of neonatal rats with hypoxic-ischemic brain damage.

The present study aimed to investigate the influence of mouse nerve growth factor (mNGF) on glial fibrillary acidic protein (GFAP) expression in neonatal rats with hypoxic-ischemic brain damage (HIBD). A total of 60 7-day-old neonatal rats were randomly divided into control, HIBD and mNGF groups (n=20). The rats in the mNGF group were injected intramuscularly with mNGF once a day for 5 days. Each group was randomly divided into a day 7 subgroup and a day 14 subgroup according to the time of sacrifice. After the rats were sacrificed, the expression of GFAP in the hippocampus in the three groups was confirmed by immunohistochemical analysis. The results revealed that the expression level of GFAP in the ischemic side of the hippocampus in the mNGF and HIBD groups was higher compared with that in the control group at days 7 and 14 after surgery, respectively (P<0.01). GFAP-positive cells were mainly distributed in the ischemic side of the hippocampal dentate gyrus (DG) region in the mNGF group while in the HIBD group they were in the ischemic side of the hippocampal CA1 region. Compared with day 7, the expression of GFAP in the ischemic side of the hippocampus in the mNGF group increased at 14 days (P<0.01), but decreased in the HIBD group (P<0.01); however, this was still higher than that in the control group (P<0.01). This study revealed that mNGF increases the expression of GFAP in the hippocampus of neonatal rats with HIBD and therefore may have a role in the repair of HIBD.

Effect of hyperbaric oxygen on neurological recovery of neonatal rats following hypoxic-ischemic brain damage and its underlying mechanism.

OBJECTIVE: To investigate the mechanism underlying the effect of hyperbaric oxygen (HBO) on hypoxic/ischemic brain damage (HIBD) in a neonatal rat model. METHODS: A total of 30 neonatal SD rats aged 7 days were randomly assigned into control group, HIBD group and HBO group (n=10 per group). Following HIBD modeling in neonatal rats, HBO treatment was performed for consecutive 7 days. Immunohistochemistry was done to measure the expression of bone morphogenetic protein-4 (BMP-4) and nestin in the hippocampus. In situ hybridization was employed to detect the mRNA expression of BMP-4 and nestin in the hippocampus. TUNEL staining was done to detect the apoptosis of nerve cells. RESULTS: HIBD was successfully established in the present study. Among three groups, the protein expression of BMP-4 in the hippocampus was the highest in the HBO group, and the smallest in the HIBD group. The BMP-4 expression in the HIBD group was significantly lower than that in the control group. The protein expression of nestin in the hippocampus was the highest in the HBO group, and the smallest in the HIBD group. The nestin protein expression in the hippocampus of HIBD group was significantly lower than that in the control group. The mRNA expression of BMP-4 in the hippocampus was the highest in the HBO group, and the smallest in the HIBD group. The mRNA expression of nestin in the hippocampus was the highest in the HBO group, and the smallest in the HIBD group. The number of apoptotic cells was the largest in the HIBD group, and the number of apoptotic cells in the HBO group was still larger than that in the control group (P<0.01). CONCLUSION: HBO may promote the neurological recovery in neonatal rats with HIBD, which may be attributed to the increased protein and mRNA expression of BMP-4 and nestin in the hippocampus and the inhibition of neural apoptosis.