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Objective:To measure the predictive value of miR-424 on the risk of sepsis complicated with acute respiratory distress syndrome(ARDS), and to explore its correlation with the prognosis of ARDS children.Methods:A prospective study was conducted.The data of children with sepsis (some complicated with ARDS), who were treated in Henan Provincial People′s Hospital (People′s Hospital, Zhengzhou University) were collected from February 2020 to February 2021.The incidence of ARDS and the fatality rate of ARDS children were recorded.Children were divided into survival group and death group according to whether death or not.The expression of miR-424 in peripheral blood mononuclear cells was detected by quantitative reverse transcription polymerase chain reaction (RT-qPCR). Pearson correlation analysis was made to investigate the relationship between the expression level of miR-424 and Pediatric Critical Illness Score(PCIS), Sequential Organ Failure Assessment(SOFA) score, oxygenation index(P/F ratio), C-reactive protein(CRP), procalcitonin (PCT), interleukin 6 (IL-6) and interleukin 8 (IL-8). The factors affecting the prognosis of ARDS children were analyzed by multivariate Logistic regression.Receiver operating characteristic curve(ROC)was used to assess the accuracy of miR-424 in early diagnosis of sepsis complicated with ARDS, and to measure the predicative value of miR-424 on the risk of death in sepsis patients with ARDS. Results:A total of 121 sepsis patients were included in this study, and 36 cases of them were complicated with ARDS.The expression level of miR-424(0.56±0.17)in the blood of sepsis patients complicated with ARDS was significantly lower than that of sepsis patients without ARDS (0.98±0.26)( t=8.776, P<0.001). The expression of miR-424 was negatively co-rrelated with IL-6( r=-0.627, P<0.01), IL-8 ( r=-0.651, P<0.01) and CRP( r=-0.472, P<0.05)in sepsis patients with ARDS.The expression of miR-424 was positively correlated with PCIS score( r=0.330, P<0.05), P/F ratio ( r=0.592, P<0.001) and albumin ( r=0.496, P<0.05) in sepsis patients with ARDS.The ROC showed that miR-424 [area under curve(AUC)=0.908, 95% CI: 0.856-0.960] could differentiate sepsis patients with ARDS from those without ARDS.Compared with that of the survival group(0.63±0.15), miR-424 of the death group decreased significantly(0.42±0.14)( t=3.890, P<0.05). The decreased miR-424 (AUC=0.845, 95% CI: 0.696-0.995)indicated an increased risk of death in children with ARDS.Multivariate Logistic regression analysis showed that miR-424( OR=0.001, P=0.033)and albumin ( OR=0.553, P=0.040) were independent risk factors for death in ARDS children. Conclusions:miR-424 can help with early diagnosis of sepsis complicated with ARDS, and can be used as a predictor of the prognosis of sepsis children complicated with ARDS.
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Objective:To measure the predictive value of miR-424 on the risk of sepsis complicated with acute respiratory distress syndrome(ARDS), and to explore its correlation with the prognosis of ARDS children.Methods:A prospective study was conducted.The data of children with sepsis (some complicated with ARDS), who were treated in Henan Provincial People′s Hospital (People′s Hospital, Zhengzhou University) were collected from February 2020 to February 2021.The incidence of ARDS and the fatality rate of ARDS children were recorded.Children were divided into survival group and death group according to whether death or not.The expression of miR-424 in peripheral blood mononuclear cells was detected by quantitative reverse transcription polymerase chain reaction (RT-qPCR). Pearson correlation analysis was made to investigate the relationship between the expression level of miR-424 and Pediatric Critical Illness Score(PCIS), Sequential Organ Failure Assessment(SOFA) score, oxygenation index(P/F ratio), C-reactive protein(CRP), procalcitonin (PCT), interleukin 6 (IL-6) and interleukin 8 (IL-8). The factors affecting the prognosis of ARDS children were analyzed by multivariate Logistic regression.Receiver operating characteristic curve(ROC)was used to assess the accuracy of miR-424 in early diagnosis of sepsis complicated with ARDS, and to measure the predicative value of miR-424 on the risk of death in sepsis patients with ARDS. Results:A total of 121 sepsis patients were included in this study, and 36 cases of them were complicated with ARDS.The expression level of miR-424(0.56±0.17)in the blood of sepsis patients complicated with ARDS was significantly lower than that of sepsis patients without ARDS (0.98±0.26)( t=8.776, P<0.001). The expression of miR-424 was negatively co-rrelated with IL-6( r=-0.627, P<0.01), IL-8 ( r=-0.651, P<0.01) and CRP( r=-0.472, P<0.05)in sepsis patients with ARDS.The expression of miR-424 was positively correlated with PCIS score( r=0.330, P<0.05), P/F ratio ( r=0.592, P<0.001) and albumin ( r=0.496, P<0.05) in sepsis patients with ARDS.The ROC showed that miR-424 [area under curve(AUC)=0.908, 95% CI: 0.856-0.960] could differentiate sepsis patients with ARDS from those without ARDS.Compared with that of the survival group(0.63±0.15), miR-424 of the death group decreased significantly(0.42±0.14)( t=3.890, P<0.05). The decreased miR-424 (AUC=0.845, 95% CI: 0.696-0.995)indicated an increased risk of death in children with ARDS.Multivariate Logistic regression analysis showed that miR-424( OR=0.001, P=0.033)and albumin ( OR=0.553, P=0.040) were independent risk factors for death in ARDS children. Conclusions:miR-424 can help with early diagnosis of sepsis complicated with ARDS, and can be used as a predictor of the prognosis of sepsis children complicated with ARDS.
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Objective:To analyze clinical factors related to nosocomial infection in children with extracorporeal membrane oxygenation(ECMO)support.Methods:General data, infection data and relevant factors in children with ECMO support in Bayi Children′s Hospital, the 7 th Medical Center of People′s Liberation Army General Hospital and Henan Provincial People′s Hospital from September 2012 to February 2020 were reviewed.Relevant factors of nosocomial infection in them were analyzed. Results:Among 163 cases, 36(22.1%) children supported with ECMO had infections during the period of ECMO, and 72 pathogenic microorganisms were detected, including 67 bacteria (33 Acinetobacter baumannii, 21 Klebsiella pneumoniae, and 6 Pseudomonas aeruginosa) and 5 fungi.Pathogens from the respiratory system, blood system, urinary tract and abdominal cavity were detected in 45 cases(62.5%), 25 cases (34.7%), 1 case (1.4%), and 1 case (1.4%), respectively.Drug sensitivity analysis of the Acinetobacter baumannii showed that it was the extensively resistant strain.Compared with uninfected children supported with ECMO, ECMO support time[(10.0±6.7) d], hospitalization[(34.0±25.3) d], hospitalization cost[(234 368±113 234) yuan], preoperative oxygenation index(52.8±23.0) and lactate value[(9.6±5.9) mmol/L]were significantly higher in nosocomial infection ones[(4.6±3.2) d, (24.3±19.8) d, (161 416±65 847) yuan, 35.6±10.4, (5.6±5.4) mmol/L] supported with ECMO (all P<0.05). There was no significant difference in the mortality between 2 groups ( P>0.05). In addition, lactate level (9.8 mmol/L) and oxygenation index (36.0±12.7) were significantly higher in died children(2.7 mmol/L, 22.1±10.4) with nosocomial infection during the period of ECMO support than those of survivors (all P<0.05). Multivariate Logistic regression analysis showed that ECMO support time( OR=7.054, 95% CI: 2.206-25.525) and preoperative lactate value( OR=2.250, 95% CI: 1.378-4.611) were independent risk factors of nosocomial infection. Conclusions:Correcting underlying diseases of ECMO supporting and shortening the duration of ECMO can reduce the incidence and mortality of nosocomial infection in children who are supported with ECMO.
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Objective:To investigate the relationship between the biochemical parameters, the pulmonary pathologic injury and the immune mechanism of severe sepsis in infant porcine, and the intervention effect of Shenfu injection.Methods:Panamanian infant porcine (2-3 months old) were divided into sham operation group (Sham group; intravenous injection of normal saline), lipopolysaccharide (LPS) induced severe sepsis model group (LPS group; intravenous injection of LPS 1 mg/kg, and continuing at 0.5 mg·kg -1·h -1 for 12 hours), and Shenfu injection intervention group (SF group; intravenous injection of Shenfu injection 10 mL/kg at the same time of modeling, twice a day) according to the random number table method, with 5 in each group. Forty-eight hours after the challenge, the changes of C-reactive protein (CRP), procalcitonin (PCT), oxygenation index (PaO 2/FiO 2), base excess (BE), blood lactate (Lac) and other biochemical indexes were detected with blood sampling; the number of neutrophils [myeloperoxidase positive (MPO +)] and their activated subsets CD11b + CD64 +, M1 macrophages (CD80 + CD64 +), CD4 + and CD8 + T cells were analyzed in peripheral blood by flow cytometry. The levels of plasma cytokines were detected by enzyme linked immunosorbent assay (ELISA). The pathological damage of lung tissue was observed by hematoxylin-eosin (HE) staining. The mRNA expression of lung injury related molecules and their receptors, chemokines, cytokines, vascular endothelial related molecules and tight junction protein were detected by reverse transcription-polymerase chain reaction (RT-PCR). Results:Compared with Sham group, the levels of CRP, PCT and Lac in LPS group significantly increased, and PaO 2/FiO 2 and BE significantly decreased. In the peripheral blood, CD80 + CD64 + macrophages, CD11b + CD64 + and MPO + neutrophils, CD4 +, CD8 + T cells and tumor necrosis factor-α (TNF-α) significantly decreased, but interleukin-10 (IL-10) and vascular endothelial cell growth factor (VEGF) significantly increased; the mRNA expressions of high mobility group protein B1 (HMGB1), receptor for advanced glycation end products (RAGE), angiopoietin 2/angiopoietin 1 (Ang2/Ang1) significantly increased, Toll like receptor 9 (TLR9), chemokines (CXCL9 and CXCL10), TNF-α, IL-27, Tek tyrosine kinase 2 (TIE2), vascular endothelial cadherin (VE-CAD) and Occludin significantly decreased in lung tissue. HE staining showed inflammatory cell infiltration and exudation in the alveolar cavity, alveoli consolidation, thickening of the alveolar interstitial layer and emphysema. Compared with LPS group, Lac in SF group significantly decreased (mmol/L: 4.2±1.0 vs. 6.3±1.1, P < 0.05), while BE and PaO 2/FiO 2 significantly increased [BE (mmol/L): -6.4±2.6 vs. -11.6±2.5, PaO 2/FiO 2 (mmHg, 1 mmHg = 0.133 kPa): 180±36 vs. 105±35, both P < 0.05]; the percentage of CD80 + CD64 + macrophages, CD11b + CD64 + and MPO + neutrophils significantly increased [CD80 + CD64 +: (7.13±2.01)% vs. (3.80±0.46)%, CD11b + CD64 +: (8.33±2.55)% vs. (2.15±0.47)%, MPO +: (21.22±2.33)% vs. (8.31±0.46)%, all P < 0.05]; the mRNA expressions of HMGB1 and RAGE decreased to some extent [HMGB1 mRNA (2 -ΔΔCT): 1.81±0.45 vs. 2.23±0.85, RAGE mRNA (2 -ΔΔCT): 6.69±3.48 vs. 11.60±6.91, both P < 0.05], the mRNA expressions of CXCL9 and TIE2 increased to a certain extent [CXCL9 mRNA (2 -ΔΔCT): 1.06±0.63 vs. 0.50±0.12, TIE2 mRNA (2 -ΔΔCT): 1.42±0.68 vs. 0.27±0.16, both P < 0.05]; the pathological damage of lung tissue were significantly alleviated. Conclusions:The increased abnormality of BE, Lac, and PaO 2/FiO 2, the immune exhausting and paralysis may be important factors leading to the fatal lung injury in infant porcine with severe sepsis. The possible mechanism is that the excessive activation of HMGB1 and its receptor RAGE, the suppression of TLR9 and corresponding chemokine and inflammatory factors lead to increased endothelial damage and decreased tight connection, which in turn induces capillary leakage. The intervention of immune disorders by Shenfu injection in the severe pneumonia accompanied by sepsis may be related to elevated M1 macrophages and activated neutrophils in the peripheral blood, inhibition of HMGB1 and its receptor RAGE mRNA expression, and elevated CXCL9 and TIE2 expression.
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Objective:To evaluate the efficacy and mechanism of continuous blood purification(CBP) in the treatment of severe sepsis in children.Methods:A total of 42 patients with severe sepsis were enrolled in the study.According to the parents undefined desire to treatment, on the basis of the same treatment condition, 30 cases in the blood purification group were treated with CBP, and 12 cases in the control group were given the routine treatment without CBP treatment.The difference of each index between the two groups at the baseline(before treatment) and 72 hours after treatment were observed.Results:The differences of heart rate, mean arterial pressure and brain natriuretic peptide before and after treatment in the blood purification group were significantly higher than those in control group( P<0.05), and the differences of oxygenation index, 24-hour average urine volume, blood urea nitrogen, creatinine and Glasgow coma score in the blood purification group were significantly higher than those in control group( P<0.05). The difference of capillary refill time, lactic acid and central venous oxygen saturation in blood purification group were significantly higher than those in control group( P<0.05). The level of white blood cell count, C-reactive protein, procalcitonin and interleukin-6 in blood purification group were significantly higher than those in control group, with all significant difference( P<0.05). There was significant difference in critical illness score between two groups after treatment and before treatment.The mortality rate of the blood purification group was significantly lower than that in control group( P<0.01). Conclusion:CBP can improve cardiovascular function, brain function, renal function, tissue perfusion, inflammation index, internal environment, and improve the score of children with severe sepsis.
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Objective@#To investigate application and safety of pediatric interfacility-transport with extracorporeal membrane oxygenation (ECMO) in China.@*Methods@#The data of 48 patients transported inter-hospital from February 2016 to May 2018 were collected from the following 4 centers: pediatric intensive care unit (PICU) of Bayi Children′s Hospital Affiliated to the 7th Medical Center of PLA General Hospital, Pediatric Hospital of Fudan University, Henan Provincial People′s Hospital and Children′s Hospital of Zhejiang University School of Medicine. The data of patients′ characteristics, ECMO mode and wean rate, and mortality were reviewed, which was further compared with the data of 57 compatible inner-hospital ECMO cases with t test, Rank sum test or chi-square test.@*Results@#All the 48 interfacility-transports were accomplished by ambulance on land, with an average transfer distance of (435±422) km. The incidence of ECMO complications was 13% (6 case), without death. There were no significant differences in lactic acid, PaO2 or SaO2 before and after transport (4.0 (2.0, 7.5) vs. 3.0 (1.5, 6.0) mmol/L, Z=-1.579, P>0.05; 112(47, 405) vs. 166(122, 240) mmHg (1 mmHg=0.133 kPa), Z=-0.104, P>0.05; 0.97±0.02 vs. 0.96±0.03, t=1.570, P>0.05). Instead, PaCO2 and pH were significantly different ((47±8) vs. (42±5) mmHg, t=2.687, P<0.05; 7.3±0.2 vs. 7.5±0.2, t=3.379, P<0.05). The total ECMO weaned rate was 73% (35/48) and the survival rate was 67% (32/48). No significant differences in demographic characteristics, ECMO mode or duration, transport distance or duration, or complications existed between the survival group and the death group (7/25 vs. 2/14, χ2=0.615, P>0.05; 4/28 vs. 2/14, χ2=0, P>0.05; (405±404) vs. (493±465) km, t=0.525, P>0.05; (5±4) vs. (5±5) h, t=0.388, P>0.05; 166 (128, 239) vs. 187(52, 405) h, Z=-0.104, P>0.05; 3/32 vs. 3/16, χ2=0.734, P>0.05). The lowest lactate value in survival group before ECMO transport was significantly lower than that in the death group ((5±5) vs. (8±6) mmol/L, t=2.151, P<0.05). There were neither significant differences in age, ECMO mode or support pattern (9/39 vs. 15/42, χ2=0.845, P>0.05; 6/42 vs. 7/50, χ2=0.001, P>0.05; 29/19 vs. 38/19, χ2=0.441, P>0.05), nor in ECMO weaned rate, survival rate or complications between interfacility-transport group and inner-hospital group (35/48 vs. 37/57, χ2=0.775, P>0.05; 32/48 vs. 35/57, χ2=0.313, P>0.05; 20/48 vs. 22/57, χ2=0.102, P>0.05).@*Conclusion@#With appropriate transport equipment and mature teams who handle problems timely during the transport, critically ill children could be safely transported to the destination with ECMO.
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Objective To discuss the mechanism of miR-24-3P in the process of acute respiratory distress syndrome (ARDS).Methods miR-24-3P mimics,miR-24-3P inhibitor and their negative controls were transfected in RAW264.7 cells respectively:The relative expression of miR-24-3P in each cell was detected.The relative expression of TNF-α,IL-6,SP1 and NF-κB were detected in each cell,and the regulation of miR-24-3P on the target gene SP1 was detected by dual luciferase reporter gene.Results The expression level of TNF-α mRNA and IL-6 mRNA in each group was statistically significant (P＜0.05).The relative expression level of SP1 mRNA and NF-κB mRNA in each group was not statistically significant (P＞0.05).The expression of SP1 in each group was statistically significant (P＜0.05).The gene analysis of dual luciferase reporter showed that the fluorescence activity was significantly inhibited after cotransfection of miR-24-3P and SP1 in HEK293 cells(P＜0.05).Conclusion miR-24-3P can play an important role in the process of ARDS by influencing the translation of SP1 gene and affecting the release of inflammatory mediators.
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Objective To detect the expression pattern of microRNA in A549 cells treated by lipopolysaccharide, study the expression of miRNA-1247-3P in A549 cells under LPS treatment and explore the possible mechanism of miRNA-1247-3P in A549 cells under LPS treatment. Methods A549 cells were divided into experimental and control groups. Immunocytochemical method and RT-PCR were used to detect the changes of SP-A and SP-C. The expression of miRNAs were detected using miRNAs array in different groups. The key miR-1247-3P was collected to detect the changes of miR-1247-3P in all groups using quantitative real-time polymerase chain reaction. Results Compared with control group, the expressions of SP-A and SP-C were significantly decreased in the experimental groups (P < 0.05). MiRNA array showed that 31 miRNAs were up-regulated and 3 miRNAs were down-regulated. Compared with control group, the expression of miR-1247-3P was significantly increased in the experimental groups (P < 0.05). Conclusion The increased expression of miR-1247-3P may play an important role in the pathogenesis of ARDS.
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BACKGROUND:Foreign scholars have researched hypoxia reperfusion in zebrafish embryos, but there is no research on c-fos gene expression and the mechanism during zebrafish cerebral hypoxia reperfusion. OBJECTIVE:To observe the zebrafish embryonic brain cel apoptosis and expression of c-fos gene in brain tissues after hypoxia reperfusion. METHODS:Zebrafish embryos were selected at 48 hours post fertilization. Neonatal hypoxia reperfusion injury was simulated by gradual y leading nitrogen (99.999%) into the device. After hypoxia treatment for 6, 12 and 24 hours, the embryos received reperfusion for 6 hours under normal oxygen concentration. The embryos in the control group received normoventilation (the dissolved oxygen concentration was about 7.0 mg/L). Acridine orange staining was performed to observe the effect of different hypoxia durations on the apoptosis of neurons in zebrafish, and then the c-fos gene expression was quantitative analyzed with real-time quantitative nucleic acid amplification detection system. And the expression level of c-fos gene was compared before and after hypoxia reperfusion. RESULTS AND CONCLUSION:A smal amount of apoptotic brain cel s could be detected in the control group, and the c-fos gene expression level was decreased;in the experimental group, the number of apoptotic cel s was increased after hypoxia for 6, 12 and 24 hours, and the gene expression after hypoxia for 6 hours was increased distinctly. The results indicate that hypoxia can increase the c-fos gene expression in brain cel s of zebrafish embryos, which may be one of the mechanisms of brain cel apoptosis increasing after hypoxia.