EGFR-TNFR1 pathway in endothelial cell facilitates acute lung injury by NF-κB/MAPK-mediated inflammation and RIP3-dependent necroptosis.

Tumor necrosis factor-α (TNFα) has emerged as a pivotal effector critically correlated with disease severity in acute lung injury (ALI). Because both the excessive activation of epidermal growth factor receptor (EGFR) and tumor necrosis factor receptor 1 (TNFR1) in sepsis-induced vasculitis are markedly diminished through EGFR tyrosine kinase inhibitor, a specific mechanism must exist to modulate TNFR1 cellular fates regulated by EGFR. Here, we demonstrated that EGFR, a specific binding partner of TNFR1, exhibited an increased NF-κB/MAPK-mediated inflammation that was governed by enhanced recruitment of TNFR-associated factor 2 (TRAF2) to TNFR1 complex I in endothelial cell (EC). Moreover, EGFR activation triggered a remarkable increase in the phosphorylation of receptor-interacting protein 1 (RIP1) and its binding with receptor-interacting protein 3 (RIP3) which led to enhanced frequency of necroptosis in complex IIb. Inhibiting the kinase of EGFR disrupted the formation of complex I and complex IIb and prevents EC from NF-κB/MAPK-mediated inflammation and RIP3-dependent necroptosis. Consistently, pharmacological inhibition of EGFR can limit the destructive effects of neutrophils activation and the hyperpermeability of lung vascular in hyperinflammation period. Collectively, we have identified EC-EGFR as a modulator of TNFR1-mediated inflammation and RIP3-dependent necroptosis, providing a possible explanation for the immunological basis of anti-EGFR therapy in sepsis-induced ALI.

Predicting risk of severe neonatal outcomes in preterm infants born from mother with prelabor rupture of membranes.

BACKGROUND: Perinatal complications are common burdens for neonates born from mother with pPROM. Physicians and parents sometimes need to make critical decisions about neonatal care with short- and long-term implications on infant's health and families and it is important to predict severe neonatal outcomes with high accuracy. METHODS: The study was based on our prospective study on 1001 preterm infants born from mother with pPROM from August 1, 2017, to March 31, 2018 in three hospitals in China. Multivariable logistic regression analysis was applied to build a predicting model incorporating obstetric and neonatal characteristics available within the first day of NICU admission. We used enhanced bootstrap resampling for internal validation. RESULTS: One thousand one-hundred pregnancies with PROM at preterm with a single fetus were included in our study. SNO was diagnosed in 180 (17.98%) neonates. On multivariate analysis of the primary cohort, independent factors for SNO were respiratory support on the first day,, surfactant on day 1, and birth weight, which were selected into the nomogram. The model displayed good discrimination with a C-index of 0.838 (95%CI, 0.802-0.874) and good calibration performance. High C-index value of 0.835 could still be reached in the internal validation and the calibration curve showed good agreement. Decision curve analysis showed if the threshold is > 15%, using our model would achieve higher net benefit than model with birthweight as the only one predictor. CONCLUSION: Variables available on the first day in NICU including respiratory support on the first day, the use of surfactant on the first day and birthweight could be used to predict the risk of SNO in infants born from mother with pPROM with good discrimination and calibration performance.

Latency period of PROM at term and the risk of neonatal infectious diseases.

To find the risk of time thresholds of PROM for infectious diseases of term neonates. A multi-center prospective cohort study including pregnancies with PROM at term with a single fetus were conducted. Time thresholds of the duration from PROM to delivery were examined in 2-h increments to assess the rates of infectious neonatal diseases. 7019 pregnancies were included in the study. Neonatal pneumonia and sepsis were most frequent infectious diseases in neonates born from mother with PROM at term. Rates of early-onset pneumonia varied significantly when comparing length of time of PROM greater than 16 h vs. less than 16 h (for EOP in 3 days of life, adjusted OR 1.864, 95% CI 1.159 ~ 2.997, p = 0.010; for EOP in 7 days of life, adjusted OR 1.704, 95% CI 1.104 ~ 2.628, p = 0.016). Neonates born from mother of whom the length of time from PROM to delivery ≥ 16 h were at a higher risk of acquiring EOP.

Treatment of neonates with respiratory distress syndrome by proportional assist ventilation plus synchronized intermittent mandatory ventilation: a comparison study.

BACKGROUND: The proportion assisted ventilation (PAV) can improve patient-ventilator interaction, reducing the incidence of end-expiratory asynchrony and increasing the time of synchrony. PAV could compensate for the leaks by elastic and resistive unloading and thus is ideal for neonates with uncuffed airways. The aim of this study was to compare the relevant clinical parameters of neonates with respiratory distress syndrome (RDS) who are supported by PAV plus synchronized intermittent mandatory ventilation (SIMV) and SIMV. METHODS: Forty-six neonates diagnosed as RDS who required mechanical ventilation were randomly divided into observer group (support by PAV+SIMV mode, N.=23) and control group (support by SIMV mode, N.=23). The X-ray grading situation, the number of asynchrony-delayed trigger, mean arterial blood pressure (MABP), spontaneous respiratory rate (RR), heart rate (HR), blood gas analysis values and circulation and respiratory parameters at each timepoint after 30 minutes, 12, 24, 48 and 72 hours of mechanical ventilation were observed. RESULTS: The forty-four neonates in two groups have been cured, the other 2 neonates (one in each group) gave up treatment and automatically discharged. There were no statistically significant differences in male, gestational age, body weight, duration of mechanical ventilation, oxygen dependence and hospital stay between the two groups (all P>0.05). There were no statistically significant differences in MABP, HR and ratio of arterial-to-alveolar partial pressure of oxygen (a/APO2) at each time point after mechanical ventilation between the two groups(all P>0.05). The number of asynchrony-delayed trigger in observer group was lower than that in control group, the difference was statistically significant (P<0.05). The number of spontaneous RR in observer group was higher than that in control group. The difference was statistically significant (P<0.05). CONCLUSIONS: The PAV plus SIMV shows a good clinical effect in treatment of neonates with RDS. It could be better to use neonatal spontaneous breathing and might cause less damage to the lung than SIMV mode.

Caffeine versus aminophylline in combination with oxygen therapy for apnea of prematurity: A retrospective cohort study.

The present study was conducted to investigate the clinical significance of caffeine and aminophylline in the treatment of premature infants with apnea under varying conditions of oxygen (O2) delivery. The clinical data of 120 premature infants with apnea treated with oxygen therapy and either caffeine citrate (20 mg/kg/day; n=77) or aminophylline (10 mg/kg/day; n=43) were retrospectively examined. The therapeutic performance of the drugs after the completion of the treatment was evaluated primarily according to the risk of recurrent episodes of apnea, the changes in the duration and concentration of inhaled O2 and the incidence of complications. In contrast to aminophylline, caffeine treatment significantly reduced the duration of O2 inhalation and the inhaled O2 concentration in the infants treated with mechanical ventilation or O2 delivery devices (P<0.05). Treatment with caffeine also decreased the incidence of recurrent apnea events and complications in the investigated population (P<0.05 or P<0.01). Caffeine performs better than aminophylline in the treatment of premature infants with apnea under different conditions of O2 delivery. The therapeutic performance of caffeine is achieved primarily via improving the efficacy of supplemental O2 and reducing the incidence of complications.

The correlation between prelabour rupture of the membranes and neonatal infectious diseases, and the evaluation of guideline implementation in China: a multi-centre prospective cohort study.

BACKGROUND: The aim of this study was to describe the epidemiology of prelabour rupture of membranes (PROM) in China and to assess the association between clinical practice following the guidelines and early neonatal infections. METHODS: We conducted a prospective cohort study of 15926 deliveries in ShenZhen Baoan Women's and Children's Hospital, Xibei Women's and Children's Hospital and Chengdu Women's and Children's Hospital between August 1, 2017, to March 31, 2018. Clinical data were collected for each participant. The epidemiology of PROM was described. The association between PROM with early neonatal infectious outcomes and the influence of the implementation of the guideline on early neonatal infectious outcomes were assessed. FINDINGS: The incidence of PROM was 18•7%. PROM was showed to be a risk factor for neonatal infectious diseases (adjusted OR 1•92, 95%CI 1•49~2•49, p<0•0001), early-onset pneumonia (EOP) (adjusted OR 1•81, 95%CI 1•29~2•53, p=0•0006) and early-onset sepsis(EOS) (adjusted OR 14•56, 95%CI 1•90~111•67, p=0•01) for term neonates. For term neonates born from mother with PROM, induction of labor according to the guideline was a protective factor for neonatal diseases(adjusted OR 0•50, 95%CI 0•25~1•00, p=0•00498) and EOP(adjusted OR 0•32, 95%CI 0•11~0•91, p=0•03). For preterm neonates born from mother with PROM, using antibiotics according to the guideline showed to be protective for neonatal infectious diseases (adjusted OR 0•14, 95%CI 0•09~0•23, p<0•0001) and EOP (adjusted OR 0•08, 95%CI 0•04~0•14, p<0•0001). INTERPRETATION: Our study showed the risk of PROM for infectious diseases (including EOP and EOS) and the benefit of the usage of antibiotics according to the guideline for infectious diseases and EOP for preterm neonates. FUNDING: National Natural Science Foundation of China, Capital Medical Development Research Fund of Beijing.

miR-206 inhibits FN1 expression and proliferation and promotes apoptosis of rat type II alveolar epithelial cells.

Bronchopulmonary dysplasia (BPD) is a syndrome of respiratory distress caused by chronic lung injury, primarily in preterm infants. miR-206 and fibronectin 1 (FN1) are associated with the development of BPD. The present study used rat type II alveolar epithelial cells (AECII) to investigate the underlying mechanisms of BPD. AECII were isolated using a primary cell culture prior to alkaline phosphatase staining and immunofluorescence of surfactant protein C (SP-C). These were used to verify the presence of AECII. AECII were then divided into four groups, which were transfected with four different plasmids. Reverse transcription-quantitative polymerase chain reaction (RT-qPCR) was performed to determine the relative expression of miR-206 in the each group. The gene and protein expression level of FN1 was detected by RT-qPCR and immunofluorescence. The proliferation of AECII in each of the four groups was evaluated using an MTT assay 48 h following transfection. The percentage of apoptotic cells was determined by flow cytometric analysis. The present study demonstrated that upregulation of miR-206 decreased the expression of FN1 (P<0.05) and low levels of miR-206 led to increased expression of FN1 (P<0.05) in AECII. Furthermore, the forced expression of miR-206 suppressed proliferation and promoted apoptosis of AECII while downregulation of miR-206 had the opposite effect (P<0.05). The results of the current study provide valuable insights into the prevention of BPD and suggest that miR-206 may be used as a potential molecular target for BPD therapy in the future.

Association between Hemoglobin Levels in the First 3 Days of Life and Bronchopulmonary Dysplasia in Preterm Infants.

Objective The objective of this study was to determine the association between hemoglobin (Hb) levels in the first 3 days of life and bronchopulmonary dysplasia (BPD) in preterm infants. Study Design The study population comprises 147 neonates with a gestational age (GA) of less than 32 weeks who were admitted to BaYi Children's Hospital Affiliated to Beijing Military General Hospital from January 2014 to May 2015. Hb levels in the first 3 days of life, maternal and infant characteristics, were recorded and then analyzed. Results BPD patients had a lower GA and birth weight than non-BPD patients. Rates of surfactant use, use of early inhalation hormone, days of mechanical ventilation > 2 weeks, and patent ductus arteriosus in BPD patients were higher and have a significant difference. Number of transfusions was higher in BPD patients. Lower Hb levels in the first 3 days of life were also observed in BPD patients. A cutoff value of Hb levels was determined as 155.5 g/L. Hb ≤ 155 g/L in the first 3 days of life was a significant risk factor for BPD. Conclusion Our study demonstrated that lower Hb levels in the first 3 days of life may increase the risk of developing BPD in preterm infants.

Association between anemia and bronchopulmonary dysplasia in preterm infants.

Anemia is commonly seen in preterm infants. It may reduce the capacity of hemoglobin to transport oxygen throughout the body and may result in tissue and organ dysfunction. This study aimed to investigate the effect of anemia on the development of bronchopulmonary dysplasia (BPD) in preterm infants. 243 infants who were admitted to BaYi Children's Hospital Affiliated to Clinical Medical College in Beijing Military General Hospital with gestational age (GA) less than 32 weeks from February, 2014 to February, 2015 were included in the study. Maternal and infant data were recorded. Multivarariate logistic regression analysis was performed to determine the association between anemia and BPD. Of 243 preterm infants, the incidence of anemia was higher in BPD patients than non-BPD patients (p < 0.001). Mean Hct in BPD patients was lower than non-BPD patients at different time points in 1d, 7d, 14d, and 21d. Controlling for other confounding factors, early anemia was associated with an increased risk of BPD. Number of transfusions is also a significant risk factor for BPD (p = 0.001). Therefore, prevention and treatment of early anemia is necessary and reducing number of transfusions may reduce the incidence of BPD in preterm infants.

[Clinical features of Candida albicans sepsis in preterm infants: an analysis of 13 cases].

OBJECTIVE: To investigate the clinical features of Candida albicans sepsis in preterm infants. METHODS: Retrospective analysis was performed on the clinical data of 13 preterm infants with Candida albicans sepsis, who were born at 28 to 36 weeks of gestational age and who weighed between 1400 and 2815 g. RESULTS: The infants were infected with Candida albicans at the age of 19±11 d, with the main clinical manifestations being apnea, poor response, poor skin perfusion, blood oxygen concentration decrease, dark skin, yellowish skin, heart rate increase in the rest state, copious phlegm and difficulty in weaning from the ventilator. The infants showed significantly decreased platelet and increased C-reactive protein (CRP), platelet distribution width (PDW), alanine transaminase (ALT), creatine kinase isoenzyme-MB (CK-MB), total bilirubin (TBIL), creatine kinase (CK), and lactate dehydrogenase (LDH). CK and LDH were significantly decreased after 2 weeks of antifungal therapy. Only 3 cases developed drug resistance to fluconazole and these showed response when treated with voriconazole instead. Of the 13 cases, 10 were cured, 2 abandoned therapy and 1 died. CONCLUSIONS: The clinical manifestations of Candida albicans sepsis are nonspecific in preterm infants. Infectious diseases are probably caused by Candida albicans in preterm infants 2-3 weeks after birth. Preterm infants show decreased platelet and increased CRP, PDW, ALT, CK-MB, TBIL, CK, and LDH when infected with Candida albicans.

Effects of different ventilation strategies on lung injury in newborn rabbits.

BACKGROUND: The results from experimental and clinical studies have shown that mechanical ventilation (MV) and/or hyperoxia may aggravate a pre-existing lung injury, or even cause lung injury in healthy lungs, despite the fact that it might be the only life-saving intervention available to a patient. The present study was designed to investigate the roles of MV and hyperoxia in the pathogenesis of lung injury. METHODS: Newborn New Zealand white rabbits were randomly assigned to an unventilated air control group or to one of the 2 × 3 × 3 ventilation strategies using a factorial design. The experimental groups were assigned different fractions of inspired oxygen (FiO(2)), peak inspiratory pressures (PIP), and respiratory times (RT). The lung wet-to-dry ratio (W/D), lung histopathology scores, and cells in the bronchoalveolar lavage fluid (BALF) were analyzed for each group. The apoptosis levels were studied by immunohistochemistry and a terminal deoxynucleotidyl transferase dUTP nick end-labeling (TUNEL) assay. RESULTS: Different ventilation regimes induced alterations in microvascular permeability, differential histopathological grading, WBC and/or neutrophil and/or lymphocyte influx, and apoptosis levels; moreover, there were significant correlations and interaction effects between these indices. CONCLUSIONS: Our data demonstrate that different ventilation regimes can induce lung injury and that the interaction effects of the FiO(2), the PIP and the RT may play crucial roles in the pathogenesis of lung injury.

A comparison study between Candida parapsilosis sepsis and Candida albicans sepsis in preterm infants.

In this study, we aimed to investigate the clinical characteristics of Candida parapsilosis sepsis in preterm infants. In this retrospective analysis of the clinical data of 11 cases of Candida parapsilosis sepsis and 13 cases of C. albicans sepsis, the two groups were compared for research using one-way analysis of variance (one-way ANOVA), chi2 test, and non-conditional logistic regression analysis. Compared to the C. albicans sepsis group, the C. parapsilosis group demonstrated a significantly lower birth weight (1331.8 +/- 252.41 vs. 1721.2 +/- 589.08) and significantly longer hospital stay (69.909 +/- 20.782 vs. 38.385 +/- 19.923) (t'/t = 2.160, -3.787; p = 0.045, 0.01, respectively); the incidences of retinopathy of prematurity (ROP) (72.7% vs. 30.8%), tienam administration (72.7% vs. 23.1%), pneumothorax, and thoracic closed drainage (27.3% vs. 0) were higher (chi2 = 4.196, 5.916, 4.052; p = 0.041, 0.015, 0.044, respectively). Logistic regression analysis indicated only hospital stay and tienam administration in the regression equation (chi2 = 18.008, p = 0.000). Compared with C. albicans sepsis, an average length hospital stay and administration of tienam are the high-risk factors for C. parapsilosis sepsis. With regard to the other predisposing factors of preterm infant fungal sepsis, there were no differences between the two groups.

[Screening and risk factors analysis of retinopathy of prematurity].

OBJECTIVE: To analyze the risk factors of retinopathy of prematurity (ROP) and provide evidence for the rational establishment of screening standard. METHODS: The clinical data of 1675 preterm infants at gestational age < or = 36 weeks or birth weight < or = 2500 g who were admitted to the neonatal intensive care unit and had been screened in our hospital from July 2006 to May 2008 were analyzed retrospectively by univariate analysis and Logistic regression analysis. Gender, birth count, gestational age, birth weight, oxygen therapy, and mother's conditions were recorded. RESULTS: ROP was detected in 195 (11.6%) of 1675 infants, of whom 35 infants (2.1%) had type 1 or threshold ROP. The lower the birth weight, the smaller the gestational age and the longer the time of oxygen therapy were, the higher the incidence of ROP was. For the infants whose birth weight was < or = 1200 g, 1201 - 1500 g, 1501 - 2000 g, 2001 - 2500 g, the incidence of ROP was 73.2%, 30.4%, 8.0%, and 1.1%; for those at gestational age < or = 30 weeks, 30(+1)-32 weeks, 32(+1)-34 weeks, 34(+1)-36 weeks, the incidence of ROP was 67.6%, 16.9%, 3.9%, and 1.0%; for the infants underwent oxygen therapy for 0 d, -3 d, -5 d, -8 d, > 8 d, the incidence of ROP was 1.5%, 3.3%, 9.6%, 23.2% and 38.8%;in the infants who inhaled oxygen at concentrations of 0.40, -0.60, -0.80 and > 0.80, the incidence of ROP was 11.8%, 18.1%, 26.8%, and 52.6%, respectively. Logistic regression analysis indicated that low birth weight, small gestational age, asphyxia, apnea, oxygen therapy were the high risk factors of ROP (the odds ratio was 0.957, 1.052, 1.186, 5.314, and 1.881). CONCLUSIONS: Low birth weight, small gestational age, asphyxia, apnea, and oxygen therapy were the high risk factors of ROP. It is recommended that all preterm infants with high risk factors should be screened.

[Construction and identification of RNA interference vector for human tissue factor gene].

OBJECTIVE: To construct a RNA interference vector for human tissue factor (TF) gene. METHODS: Human TF short hairpin RNA (shRNA) sequence was designed using online design software (Invitrogen) and synthesized into double-strand oligonucleotide (ds oligo), which was cloned into the pENTRTM/U6 plasmid, followed by transformation of the product into competent Top10 E. coli cells. After expansion of the transformed bacteria, the plasmid was extracted and sequenced, which was subsequently transfected into human umbilical vein endothelial cells (HUVECs). The interference effect of the vector on the target gene expression was detected by RT-PCR and immunofluorescence assay. RESULTS: The sequencing result indicated that the plasmid pENTRTM/U6-RelB-shRNA was constructed correctly, which resulted in effective inhibition of TF expression in HUVECs after transfection. CONCLUSION: The RNA interference vector against human TF gene has been constructed successfully, which may provide a stable transfection vector for potential treatment of blood coagulation abnormalities.