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Objective:To observe the therapeutic effect of echinacoside (ECH) on liver injury and glucose metabolism disorder in sepsis rats induced by cecal ligation and puncture (CLP), and to explore its possible mechanism.Methods:Forty eight male Sprague Dawley (SD) rats were randomly divided into four groups: sham group (sham), model group (CLP), treatment group (CLP+ ECH) and inhibitor group (CLP+ ECH+ EX527). The sham group only received laparotomy, and the model group underwent CLP. The treatment group was intragastric administration of echinacea (30 mg/kg) every day after CLP modeling. The inhibitor group was injected with silence information regulator 1 (SIRT1) inhibitor EX527 (5 mg/kg) one hour before CLP, and then treated the same as the treatment group. Fasting blood glucose, insulin and serum biochemical indexes were detected in virous groups. The serum levels of interleukin (IL)-1 β, IL-6 and tumor necrosis factor-α(TNF-α) were detected by enzyme-linked immunosorbent assay (ELISA). 2′, 7′- dichlorofluorescein diacetate (DCFH-DA) staining was used to observe the production of reactive oxygen species (ROS) in liver tissue of rats in each group; hematoxylin-eosin (HE) staining was used to observe the pathological changes of liver tissue in each group; The expressions of SIRT1, glucose-6-phosphatase (G6Pase), phosphoenolpyruvate carboxykinase (PEPCK), phosphorylated signal transducers and activators of transcription 3 (p-STAT3) and phosphorylated protein ki-nase B(p-AKT) were detected by Western blot.Results:Compared with sham group, the levels of serum glucose, serum insulin, alanine aminotransferase (ALT), aspartate aminotransferase (AST), ROS, IL-1β, IL-6 and TNF-α in model group increased, while the liver glycogen and survival rate decreased (all P<0.05). After echinacoside treatment, the serum glucose, serum insulin, ALT, AST, ROS , IL-1β, IL-6 and TNF-α levels decreased, and the liver glycogen and survival rate increased (all P<0.05); After SIRT1 inhibitor intervention, the levels of serum insulin, ALT, AST, IL-6 and ROS in the inhibitor group increased ( P<0.05). HE staining showed that there were infiltration and necrosis of inflammatory cells in the liver tissue of model group, and echinacoside could significantly reduce the focal and massive necrosis; Western blot showed that compared with the sham group, the expression levels of SIRT1, p-STAT3 and p-AKT protein in the model group decreased, while the expression levels of G6Pase and PEPCK protein increased ( P<0.05); After echinacoside treatment, the expression levels of SIRT1, p-STAT3 and p-AKT increased, while the expression levels of G6Pase and PEPCK decreased ( P<0.05). After SIRT1 inhibitor intervention, the expression of SIRT1, p-STAT3 and p-AKT protein decreased, and the expression of G6Pase and PEPCK protein increased in the inhibitor group ( P<0.05). Conclusions:Echinacoside is a potential therapeutic agent for sepsis associated liver injury and glucose metabolism disorders, which may play a role by targeting SIRT1 to activate STAT3 and AKT in the liver.
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Objective:To investigate whether miRNA-296-5p can inhibit enterovirus 71 (EV71) virus replication in neural cells SK-N-SH by targeting the phosphatase and tensin homology deleted on chromosome 10 (PTEN)/phosphatidylinositol 3-kinase (PI3K)/serine/threonine kinases (AKT) signaling pathway.Methods:Serum samples were collected from patients with EV71 virus-infected hand-foot-mouth disease and normal physical examination, and the expressions of serum inflammatory factors procalcitonin (PCT), C-reactive protein (CRP), tumor necrosis factor-α (TNF-α) , interleukin-6 (IL-6), IL-1β, and IL-13 were detected by enzymelinked immunosorbent assay (ELISA). Human neuroblastoma SK-N-SH cells infected by EV71 virus in logarithmic growth phase were set up as control group, miRNA-296-5p mimic group and miRNA-296-5p inhibitor group. The transfection was carried out according to the Lipofectamine 2000tm cell transfection reagent. The expression of EV71-VP1 gene mRNA and protein and PTEN/PI3K/Akt signal pathway related molecules in three groups of cells was observed by real-time fluorescent quantitative polymerase chain reaction (qRT-PCR) and Western blot.Results:ELISA test results showed that the levels of serum inflammatory factors PCT, CRP, TNF-α, IL-1β, IL-6 and IL-13 in patients with EV71 virus-infected hand, foot and mouth disease were significantly higher than those in normal physical examination ( P<0.05). The results of qRT-PCR and Western blot showed that the mRNA and protein levels of mirna-296-5p and PTEN in SK-N-SH were significantly decreased after EV71 virus infection, while the mRNA and protein levels of EV71-VP1 and molecules related to PI3K/AKT signaling pathway were significantly increased ( P<0.05). The expression of PTEN was significantly increased in the miRNA-296-5p mimic group, and the expression of EV71-VP1 and the activation of the PI3K/AKT signaling pathway were inhibited, while the effect was reversed in the miRNA-296-5p inhibitor group ( P<0.05). Conclusions:MiRNA-296-5p inhibits the replication of EV71 virus in neural cells SK-N-SH by targeting the PTEN/PI3K/AKT signaling pathway, while reducing the cellular inflammatory response, targeting miRNA-296-5p and downstream PTEN/PI3K/AKT The signal pathway is expected to provide therapeutic targets and theoretical basis for the treatment of hand-foot-mouth disease caused by clinical EV71 virus.
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To analyze the clinical manifestations and gene mutations in children with congenital insensitivity to pain with anhidrosis (CIPA), and review related literature. An infant diagnosed with congenital insensitivity to pain with anhidrosis was reported. The main clinical manifestations of the infant were painless, no sweat, and repeated fever. Peripheral blood of the infant and his parents was collected, and candidate variants were confirmed by Sanger sequencing. The results of molecular genetic analysis showed that there were compound heterozygous mutations (c.36G>A, c.851-33T>A) of neurotrophic tyrosine kinase receptor type 1 (NTRK1) in the infant. c.36G>A and c.851-33T>A were inherited from his father and mother, respectively. c.851-33T>A is a previously reported mutation, c.36G>A is an unreported mutation, which can lead to the tryptophan changing into a stop codon. According to the American College of Medical Genetics and Genomics (ACMG) variant interpretation guidelines, the mutation is interpreted as pathogenic, and the biological hazard is potentially harmful. Congenital insensitivity to pain with anhidrosis is a rare inherited disorder. Genetic molecular genetic analysis is helpful to diagnose and discover new gene mutations.
Subject(s)
Humans , Infant , Channelopathies , Mutation , Pain Insensitivity, Congenital , Receptor, trkAABSTRACT
Objective@#To analyze the clinical features of acute liver failure in critically ill children, to investigate the risk factors affecting the prognosis of acute liver failure in critically ill children, and to provide evidence for clinical diagnosis and prognosis.@*Methods@#The data of 85 children with acute liver failure who conformed to the diagnostic criteria from January 2011 to January 2017 at Hunan Children′s Hospital were analyzed.According to the prognosis of children with acute liver failure, all children were divided into the survival group (49 cases) and the death group (36 cases). Univariate and multivariate Logistic regression methods were used to analyze the risk factors affecting the prognosis of children with acute liver failure.@*Results@#There were statistically significant differences of enzyme bile isolation[26.5% (13/49 cases) vs.55.6%(20/36 cases), χ2=7.361, P=0.007], hepatic encephalopathy [16.3%(8/49 cases) vs.38.9%(14/36 cases), χ2=5.507, P=0.019], total bilirubin [72.30(3.93-428.80) μmol/L vs.153.08(3.23-776.26) μmol/L, U=-2.283, P=0.027], albumin [35.02(22.89-45.30) g/L vs.28.90(18.30-40.26) g/L, U=4.640, P=0.000], alanine aminotransferase [1 626.10(23.01-9 518.41) U/L vs.533.08(7.02-5 163.83) U/L, U=2.992, P=0.004], aspartate aminotransferase [1 611.20(34.51-15 850.02) U/L vs.715.54(60.06-10 280.42) U/L, U=2.312, P=0.023], blood ammonia [71.02(16.04-148.56) μmol/L vs.81.02(33.04-274.02) μmol/L, U=-2.057, P=0.046], prothrombin time [23.10(13.61-81.23) s vs.33.91(12.62-84.57) s, U=-2.364, P=0.022], activated partial prothrombin time [52.91(4.02-181.02) s vs.67.35(31.20-180.02) s, U=-2.226, P=0.029], and end-stage liver disease model score [13.00(-9.00-52.00) score vs.27.50(-9.00-88.00) score, U=-3.082, P=0.003] in the children with acute liver failure between the survival group and the death group.Multivariate Logistic regression analysis showed that total bilirubin (OR=0.236, 95%CI: 0.059-0.946, P=0.041), albumin(OR=0.854, 95%CI: 0.746-0.976, P=0.019), enzyme bile separation (OR=1.063, 95%CI: 1.031-1.119, P=0.005) and hepatic encephalopathy (OR=1.439, 95%CI: 1.021-2.043, P=0.017) were risk factors for the prognosis of children with acute liver failure.@*Conclusions@#The level of the total bilirubin and the ratio of enzyme bile isolation and hepatic encephalopathy are positively correlated but the level of albumin is negatively correlated with the prognosis of critically ill children with acute liver failure.
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Objective To comment the severity of severe hand,foot and mouth disease(HFMD)by pediatric risk of mortality score(PRISM),and assess the performance of PRISM in predicting mortality or complication probability in HFMD.Methods Four hundred and twenty-four severe HFMD pediatric patients were recruited in the study from 1th Jan 2010 to 31th June 2013.Information on the outcome and the varia-bles required to calculate PRISM score were collected.The logistic regression model developed in the learning sample was evaluated in the test sample by calculating the area under the receiver operating characteristic (ROC)curve to assess discrimination pneumorrhagia and death.Calibration across deciles of risk was evalua-ted using the Hosmer-Lemeshow goodness-of-fit χ2 test.Results The area under the ROC curve were 0.87 (95%CI 0.80~0.94 )for PRISM in predicting pneumorrhagia probability.The area under the ROC curve were 0.87(95%CI 0.80~0.95)for PRISM in predicting mortality probability.The PRISM in observed and expected pneumorrhagia did not demonstrate good calibration at ten mortality risk intervals (χ2 =36.66, P<0.001 ).The PRISM in observed and expected mortality did not demonstrate good calibration at ten mortali-ty risk intervals(χ2 =41.11,P<0.001).Conclusion The PRISM score is demonstrated good discrimination of pneumorrhagia and death in HFMD pediatric patients,but the performance of calibration is not good.
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Objective To investigate the serum procalcitonin (PCT) levels in sepsis caused by the bacteria,virus and mycoplasma and explore the role of PCT in etiology diagnosis of sepsis in children.Methods Three hundreds and thirty critically ill children with sepsis caused by bacteria,virus and mycoplasma admitted in PICU of Hunan Children' s Hospital from Feb 1,2011 to Sep 1,2012 were reviewed and analyzed.The PCT levels were measured at admission and day 3.The differences in accidence of sepsis caused by bacteria,viruses and mycoplasma according to different serum PCT levels were analyzed.The differences of PCT levels at admission and day 3 in sepsic children caused by bacteria,viruses and mycoplasma were analyzed.Results The level of serum PCT in sepsis caused by bacterial infection were distinctly increased,caused by virus and mycoplasma infections was not obvious but the increases of serum PCT [0.71 (8.14)ng/ml,0.15 (1.68) ng/ml,0.28 (1.89) ng/ml].According to various PCT levels(0.05 ～ ng/ml,0.5 ～ng/ml,2 ～ ng/ml,10 ～ 300 ng/ml),the differences of accidence of sepsis caused by bacteria,virus and mycoplasma were also statistically significant(x2 =84.50,P ＜ 0.01).The PCT level of septic children caused by bacterial infection in day 3 was significantly decreased compared with that at admission [0.32 (5.68) ng/ml vs 0.71 (8.14) ng/ml] (U =19.34,P ＜0.05).Conclusion PCT plays a certain role in etiology diagnosis of sepsis in children.The increased PCT levels which can be reduced by anti-inflammatory treatment indicate the likelihood of bacterial infection and sepsis.The increase of PCT induced by viral and mycoplasma infections is not obvious,but bacterial infection can not be completely ruled out.
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Objective To discuss the signiifcance of serum albumin level in assessing severity, progress and prognosis of sepsis in children. Methods The clinical data of 212 patients diagnosed with sepsis admitted to PICU from February 2010 to July 2010 were retrospectively analyzed, and 52 patients had severe sepsis and 31 patients had septic shock. Meanwhile, 110 non-sepsis patients were selected as controls. The relationships of hypoalbuminemia with pediatric critical illness score (PCIS), pediatric risk of mortality III (PRISM III) and prognosis were evaluated, and the change of albumin level in patients with dif-ferent severity of sepsis was observed. Relative factors analysis of albumin level ≤25 g/L was performed. Results As the serum albumin level was decreased, the PCIS was signiifcantly decreased while the PRISM III was increased (P<0.01). The se-rum albumin level was signiifcantly different among children with septic shock, severe sepsis and sepsis and controls (F=13.938, P=0.000). The results of relative factors analysis showed that sepsis children with an albumin level≤25 g/L had more organ failures, higher mortality, longer hospital and PICU stay and more likelihood for ventilator support (P<0.01). Lower albumin levels were accompanied with lower rates of recovery and improvement but higher mortality (rs=-0.161, P=0.000). Conclusions Hypoalbuminemia can be used as indirect indicator for severity of infection. The albumin level≤25 g/L indicated the severity of illness and prognosis in children with sepsis.