Neutrophil gelatinase-associated lipocalin(NGAL) is a new member of lipid protein family,which is a kind of stable polypeptide firstly separated from neutrophils.As a new biomarker of acute kidney injury(AKI) for early clinical diagnosis.NGAL is less easily affected by factors such as the body itself,drugs or aspects outside kidney.The high levels of NGAL in the blood and urine can both predict AKI.Dynamic monitoring NGAL can assess the validity of clinical therapy and give reference to treatment of AKI.

The invasive fungal infection( IFI)in PICU has increased steadily during the recent years. Candida spp. and Aspergillus spp. are the most frequently fungi in children. Candida spp. is the leading cause and invasive Candida spp. Infection( ICI)is approximately five times frequency than invasive Aspergillus spp. Infection( IAI). The attributable mortality of ICI or IAI remains different mainly because of different basic diseases. Stay in the PICU presents risk factors for ICI especialy using central venous catheter,parenteral nutrition,dialysis,mechanical ventilation,and prolonged antibiotics application. The patients with hematologic malignancies and leukemia are higher prevalence of IAI who were treated with cytotoxic or immunosuppres-sive drugs,broad-spectrum antibiotics and stem cell transplantation. The most important challenge remains to propose targeted prophylaxis and to identify IFI earily in high risk critically ill children in PICU.

Adrenal crisis is a life-threatening emergency caused by the destruction or altered function of the adrenal gland with a primary deficit in cortisol secretion(primary adrenal insufficiency)or by hypotha-lamic-pituitary pathologies determining a deficit of adrenocorticotropic hormone(secondary adrenal insuffi-ciency).Infection and abrupt end glucocorticoid treatment are the major precipitating causes of adrenal crisis. Patients with adrenal crisis typically present with hypovolemic shock or hypotension,nausea,vomiting,and fe-ver responding well to parenteral hydrocortisone administration.The main laboratory findings include lower serum cortisol concentrations,hyponatremia,hypoglycaemia and/or hyperkalemia.Delay diagnosis of adrenal insufficiency leads to adrenal crisis which is potentially lethal complication.Empirical glucocorticoid replace-ment should be initiated as soon as the suspicious of adrenal crisis,or sooner if the patient presents in adrenal crisis in critically ill children.

Sepsis is the leading cause of acute kidney injury (AKI) in pediatric intensive care unit.Development of AKI during sepsis increased patient morbidity,predicts higher mortality and days of stay in the intensive care unit.The mechanisms behind AKI in sepsis remain controversial but were believed to be complex and multi-factorial.The pathophysiology of AKI in sepsis involved intrarenal hemodynamic changes,endothelial dysfunction,infiltration of inflammatory cytokines.The new markers of neutrophil gelatinase-associated lipocalin as the representative is helpful for early diagnosis of AKI.Renal replacement therapy (RRT)is the main treatment of sepsis related AKI.At present,the model,dose and exact timing of RRT is not well defined.A widely accepted viewpoint is that the injury stage of RIFLE diagnostic criteria and fluid overload up to 10％ ～ 20％ is the beginning of the most appropriate chance of RRT.

Objective To investigate the effects of microRNA-155 (miR-155) inhibitor on JAK/STAT1 (Janus kinase/signal transducer and activator transcription 1) signaling pathways in the injured lung tissue induced by lipopolysaccharide (LPS).Methods One hundred and twenty BALB/c mice were randomly divided into control group (n =40),LPS group (n =40),and inhibitor + LPS group (n =40).LPS group and inhibitor + LPS group were made by injection of LPS 20 mg/kg intra-peritonealy,whereas equivalent volume of normal saline was given instead in the control group.The 80 mg/kg of miR-155 inhibitor was injected into caudal vein 24 h before LPS injection in inhibitor + LPS group.Mice were sacrificed at 6 h,12 h,24 h,and 48h separately after LPS injection,and lung tissue were collected.The levels of tumor necrosis factor-α (TNF-α) and interleukin-10 (IL-10) of lung tissue were measured using the enzyme-linked immunosorbent assay (ELISA).Using histopathological examination,the injury of lung tissue was evaluated.The expressions of miR-155,STAT1 mRNA,SOCS1 mRNA in lung tissue were assayed by fluorescent quantitative reverse transcription polymerase chain reaction (qRT-PCR).Results The miR-155 expression induced by LPS increased at 6 h,12 h,24 h and decreased at 48 h.The miR-155 expressions in LPS group were (8.52 ± 1.12) at 6 h,(11.04 ±0.99) at 12 h,(15.84 ±0.80) at 24 h and (4.03 ± 2.55) at 48 h.In the inhibitor + LPS group,the expressions of miR-155 were lower compared with LPS group,showing significant differences at 12 h (t =6.08,P ＜ 0.01),and at 24 h (t =23.64,P ＜ 0.01).STAT1 mRNA and SOCS1 mRNA both reached peak levels at 6 h after LPS injection.The levels of STAT1 mRNA in LPS group were higher than those in inhibitor + LPS group,showing significant differencesat6h (t=4.41,P＜0.01),12h(t=2.69,P＜0.05),and24h (t=3.62,P＜0.01).The levels of SOCS1 mRNA in inhibitor + LPS group were higher than those in LPS group,showing significant differences at 6 h (t =4.55,P ＜0.01),12 h (t =4.12,P ＜0.01),24 h (t =2.38,P ＜ 0.05).TNF-α reached its peak value at 6 hours and IL-10 reached its peak value at 48 hours.Both TNF-α and IL-10 were higher in LPS group than those in inhibitor + LPS group showing significant differences at 6 h,12 h,24 h (P ＜0.01).The pathologic examination indicated the lung injury in inhibitor + LPS group was milder than that in LPS group.Conclusion The miR-155 increased in lung tissue of endotoxemic mice.miR-155 inhibitor may suppress JAK/STAT1 signaling pathway and protect the lung tissue.

Objective To explore the protective effect of rnicroRNA (miRNA)-155 inhibitor on interleukin-1 receptor-associated kinase (IRAK)-1 mRNA and IRAK-4 mRNA in endotoximia induced liver injury in mice.Methods One hundred and twenty male BALB/c mice were randomly divided into healthy control group(n =40),endotoximia group (n =40) and miRNA-155 inhibitor group (n =40).Each group were divided into 6 h,12 h,24 h,48 h subgroups,each of which consisted of 10 mice.The mice in miRNA-155 inhibitor group were administered with miRNA-155 inhibitor[80 mg(kg ·d)] via tail vein injection before lipopolysaccharide (LPS) administration while the other two groups treated with normal saline,following 24 hours,model of endotoximia mice was produced by injection of LPS intraperitoneally.At 6 h,12 h,24 h,48 h after LPS exposure,the experimental mice were sacrificed and the liver tissue samples were collected.Histopathological changes,the expression of miRNA-155,IRAK-1 mRNA,IRAK-4 mRNA,tumor necrosis factor (TNF)-α,IL-1,IL-10 were detected.Results LPS exposure resulted in increase of miRNA-155,IRAK-1 mRNA,IRAK-4 mRNA,TNF-α,IL-1 and IL-10 in both endotoximia group and miRNA-155 inhibitor group compared to the control group,miRNA-155 inhibitor resulted in decrease of miRNA-155,IRAK-1 mRNA,IRAK-4 mRNA,TNF-α,IL-1 and IL-10 in miRNA-155 inhibitor group compared to the endotoximia group.There were significant differences of miRNA-155 expression at 12 h,24 h,48 h after LPS exposure among 3 groups (P ＜ 0.05).Both IRAK-1 mRNA and IRAK-4 mRNA showed significant differences at 12 h,24 h,48 h.Turning to inflammation factors,differences were found among 3 groups at all time points (P ＜ 0.05).At light-scope,there was improvement in sepsis associated liver injury in miRNA-155 inhibitor group compared to endotoximia group.Conclusion miRNA-155 inhibitor administration appears to down regulate IRAK-1 mRNA and IRAK-4 mRNA expression and further deduce the excessive inflammatory and anti-inflammatory reaction,which may alleviate liver injury in endotoximia mice.

Objective To evaluate the value of urine neutrophil gelatinase-associated lipocalin (uNGAL) to early diagnose acute kidney injury(AKI) of critically ill children in PICU.Methods Eighty critically ill children at PICU of Children's Hospital Affiliated to Shanghai Jiaotong University were enrolled in this study from April to June 2013.They were continuously observed for 72 hours.According to pediatric RIFLE criteria for diagnosis of AKI,patients were divided into AKI group (15 cases) or non-AKI group (65 cases).Additionally,according to sepsis diagnostic criteria,patients were divided into sepsis group (31 cases) or non-sepsis group (49 cases).The levels of serum creatinine and uNGAL were measured within 6th hour,24th hour,48th hour,72th hour after admitted to PICU.The differences of uNGAL levels between AKI and non-AKI groups,sepsis without AKI and non-sepsis non-AKI groups,sepsis merged AKI and sepsis without AKI groups were analysed.The sensitivity and specificity of uNGAL and serum creatinine for diagnosis of AKI at 48th hour were evaluated by ROC curve.Results Thirteen cases of eighty children developed to AKI after admitted to PICU.(1)The uNGAL levels [M(QR),ng/ml] in AKI group within 6th hour,at 24th hour,48th hour,72th hour were 863.00 (696.00),700.50 (580.00),365.50 (285.00),289.50 (319.30),respectively,which were significantly higher than those in non-AKI group [20.00 (106.00),20.00 (85.30),20.00(101.00),20.00(36.00)] (P ＜0.01).(2)The uNGAL levels in new developed group were much higher than those in non-AKI group at each time point.The comparision of serum creatinine at 48th hour was statistic difference.(3)The uNGAL levels rised at early stage in sepsis without AKI group and down to normal gradually after 48th hour.(4)The uNGAL levels continued increasing in sepsis merged AKI group,and had significant differences comparing with sepsis without AKI group(P ＜ 0.01).(5) The areas under ROC curve of uNGAL and serum creatinine at 48th hour were 0.902(95％ CI:0.801 ～ 1.004) and 0.801 (95％ CI:0.768 ～ 0.981),respectively.Conclusion The level of uNGAL has earlier increase for 24 to 48 hours than that of serum creatinine in critically ill children,and it can also reflect the severity of AKI.Therefore it can be used as an early diagnostic biomarker for AKI in PICU.

Objective To investigate the effects of lipopolysaccharide (LPS) on the expressions of sodium taurocholate co-transporting polypeptide (Ntcp) and bile salt export pump (Bsep),as well as the liver function markers in the serum including total bilirubin (TBIL),total bile acids (TBA),alanine aminotransferase (ALT),aspartate aminotransferase (AST) in mice.Methods One hundred and twenty-eight C57BL/6 mice were intra-peritoneally injected with different doses of 5,10,20 or 40 mg/kg LPS (n =24),respectively.No treatment or treated with 0.9％ NaC1 in mice as controls.Serum TBIL,TBA,ALT and AST levels were measured at 24 h,48 h and 72 h after LPS injection in each group.The mRNA expressions of Ntcp and Bsep were detected by reverse transcription quantitative polymerase chain reaction (RT-qPCR).The liver histological sections were stained with haematoxylin and eosin (H&E).Results The Ntcp and Bsep mRNA expressions in mice liver were significantly lower in livers of LPS-treated mice within 24-72 h compared with control group,and the lowest level was reached at 24 h in a dose-dependent manner.And the relative expressions of Ntcp mRNA and Bsep mRNA were (0.64 ± 0.02),(0.53 ± 0.14),(0.25±0.09),(0.15±0.07)and (0.74±0.12),(0.58±0.11),(0.41±0.09),(0.27 ± ± 0.11) in livers of mice injected with LPS in the different doses of 5,10,20,40 mg/kg,respectively.In addition,serum levels of TBIL,TBA,ALT,and AST were significantly increased in mice of LPS-treated group compared with control group,particularly within 24 h after LPS treatment.Serum levels of TBIL,TBA,ALT,and AST were significantly decreased in mice of 40 mg/kg LPS-treated 72 h group compared with 24 h group presenting them with (1.29 ± 0.25) μ mol/L vs.(1.71 ± 0.22) μ moL/L,(6.97 ± 0.98) μmol/Lvs.(8.96±1.01) μmol/L,(120.17±21.08) U/L vs.(179.22±16.57) U/L,(360.34 ±35.31) U/L vs.(510.97 ± 34.70) U/L,respectively.Furthermore,histological changes in liver depend on dose and the course of LPS treatment.Cytoplasm rarefaction and inflammatory cells infiltration were detected at 24 h after treatment with 5 or 10 mg/kg LPS.Acidophilic and vacuolar degeneration,neutrophils infiltration in the hepatic sinusoid and portal area,the proliferation of bile ductulus were observed at 48 h,72 h after treatment with 5 or 10 mg/kg LPS.In the 20 or 40 mg/kg LPS treatment groups,focal necrosis,infiltration with inflammatory cells,proliferation of bile ductulus and expansion of duct were observed at 24 h,48 h and 72 h after LPS treatment.Conclusions LPS decreases the mRNA expressions of Ntcp and Bsep in a dose dependent manner in mice,contributing to mechanism of liver injury induced by endotoxin.

Objective To investigate the effects and underlying mechanisms of methylprednisolone (MP) on liver injury induced by lipopolysaccharide (LPS).Methods Total of 48 C57BL/6 mice (8-week old) were randomly divided into the control group,LPS-induced endotoxemia model (1 h,2 h,4 h,8 h,24 h,48 h) and intervention group with MP therapy (n =6).Mice were intraperitoneally injected withLPS (20 mg/kg) for indicated time (1 h,2 h,4 h,8 h,24 h,48 h),and MP (20mg/kg) was intraperitonealinjected into micetointervene LPS-induced liver injury.Saline was used as control.Pathological changes of liver tissues were analyzed by hematoxylin & eosin (HE) staining.The serum levels of ALT,TBIL and TBA were determined,and the mRNA levels of TNF-α,IL-6,IL-1β and the protein levels of P62,LC3 Ⅱ/Ⅰ in livers were detected by real time-PCR and Western-blot.Results (1) MP therapy protects mice against LPS-induced liver injury at the dose of 20 mg (kg · d).The pathological sections showed that the structure of hepatic lobule,the hepatocyte vacuolar degeneration,eosinophilic degeneration were improved in LPS + MP/group compared with LPS group;(2) The serum levels of ALT,TBIL,TBA in LPS + MP group was significantly decreased compared with LPS 48 h group [(63.40 ±11.55) vs.(104.50±29.34) U/L,(0.37 ±0.08) vs.(0.52 ±0.12) μmol/L,(4.67 ±2.58) vs.(10.33 ± 2.34) μmol/L,P =0.009,P =0.032,P ＜ 0.01];(3) The mRNA levels of TNF-α,IL-6,IL-1β in LPS + MP group was significantly lower than that of LPS 48 h group [(4.18 ±0.81) vs.(10.09 ±4.73),(0.31 ±0.14) vs.(1.06 ±0.68),(0.17 ±0.05) vs.(1.22 ±0.50),respectively,all P ＜0.05];(4) LPS activated autophagy within 2h after LPS treatment.Then,autophagy was suppressed from 2h to 24h after LPS treatment indicated as the decreased expression of LC3 Ⅱ/Ⅰ.Interestingly,MP treatment significantly reversed LPS-suppressed autophagy showing that the protein level of LC3 Ⅱ/Ⅰ was significantly increased in LPS + MP group compared with LPS 48 h group.Conclusions MP therapy protects mice against LPS-induced liver injury and inflammation,partially due to activation of autophagy in livers.

Objective: To investigate the effects of methylprednisolone on STAT3-ERK1/2 signaling pathway in lipopolysaccharide (LPS)-induced acute lung injury (ALI). Methods: The C57BL/6J male mice (8-week-old) were randomly(random number) divided into 4 groups: control group (control), LPS-induced endotoxemia model (LPS), only methylprednisolone (MP) administration group (MP), and intervention group with 2 mg/kg MP (LPS+MP) (n= 8 per group). The wet/dry (W/D) weight ratio of lung tissue, lung pathology by hematoxylin & eosin (HE) staining, serum and mRNA levels of TNF-α and IL-6 in lungs were determined. The protein levels of p-STAT3 and p-ERK1/2 in lungs were detected by Western blot. Statistical analyses were performed using One-way analysis of variance test to compare among multiple groups. Results: (1)MP treatment significantly decreased the lung W/D weight ratio compared with the LPS group[(3.01±0.84) vs(3.87±0.17), P = 0.038]; (2) The histopathological lesions of the lung were improved in the LPS+MP group compared with the LPS group accompanied with reduced inflammatory cell infiltration and attenuated the alveolar wall thickening; (3) The serum levels of TNF-α and IL-6 in the LPS+MP group was significantly decreased compared with the LPS group[(3.17±1.64) pg/mL vs (6.61±1.27) pg/mL, P = 0.003; (1.42±0.35) pg/mL vs (3.80±1.35) pg/mL, P = 0.008, respectively], and the mRNA levels of TNF-α and IL-6 in the LPS+MP group were significantly lower than those of the LPS group [(5.10±0.81) vs (12.2±5.05), P = 0.03; (1.62±1.00) vs (11.12±6.56), P=0.026; respectively]; (4) MP therapy significantly inhibited P-STAT3 and P-ERK1/2 protein levels [(0.26±0.05) vs (0.86±0.06), P < 0.001, (0.24±0.02) vs (1.34±0.32), P < 0.001]. Conclusions Methylprednisolone protects LPS-induced acute lung injury possibly via suppressing STAT3-ERK1/2 signaling pathway and reducing TNF-α and IL-6 expression.