Objective To investigate the effect of vasoactive intestinal peptide (VIP) and methylprednisolone (MP) on Toll-like receptor (TLR)2/4 mRNA expression in endotoxin (lipopolysaccharide,LPS) induced shock.Methods Ninety Sprague-Dawley rats were randomly divided into LPS group (n =20),LPS + VIP group (n =20),LPS + MP group (n =20),LPS + VIP + MP group (n =20) and control group (n =10).LPS group injected intravenously LPS (E Coli O55B5) 10 mg/kg.LPS + VIP group,LPS + MP group and LPS + VIP + MP group were injected intravenously VIP 5 nmol/kg,MP 3 mg/kg and VIP 5 nmoL/kg + MP 3 mg/kg after LPS 10 mg/kg injection.The control group injected normal saline intravenously instead of LPS.The rats were sacrificed at 6 h and 24 h after injection and the intestine samples were collected.Pathological changes of the intestine were observed by microscopy.RT-PCR was used to detect the intestinal TLR2 mRNA and TLR4 mRNA expressions.Results Intestinal mucosa showed edema or necrotic change with structure of the microvilli disappeared after LPS injection.The inestinal lesions in VIP,MP and VIP + MP groups were milder than LPS group.At 6 h after LPS injection,TLR2 mRNA and TLR4 mRNA expressions were significantly up-regulated in LPS group,LPS + VIP group,LPS + MP group and LPS + VIP + MP group (TLR2 mRNA:1.14 ±0.38,1.17 ±0.42,1.16 ±0.41,0.92 ± 0.29;TLR4 mRNA 1.21 ±0.18,1.04 ± 0.38,1.11 ± 0.34,1.01 ± 0.20) compared with the control group (0.32 ± 0.20,0.24 ± 0.17) (P ＜ 0.01).But there was no significant difference between LPS group,LPS + VIP group,LPS + MP group and LPS + VIP + MP group (P ＞ 0.05).At 24 h after LPS injection,TLR2 mRNA and TLR4 mRNA expressions in LPS + VIP group,LPS + MP group and LPS + VIP + MP group (TLR2 mRNA:0.63 ± 0.12,0.59 ± 0.13,0.52 ±0.19;TLR4 mRNA 0.67 ±0.09,0.64 ±0.09,0.51 ±0.13) were significantly lower than LPS group (1.04 ± 0.38,0.82 ±0.18) (P ＜0.01) (P ＜0.05).Conclusion VIP and/or MP can mitigate intestinal injury induced by LPS shock.The gastrointestinal protection of VIP and glucocorticoids were related to downregulation signaling TLR2 mRNA and TLR4 mRNA expression.But VIP/MP and VIP + MP have no significant effect on expression of intestinal TLR2/4 mRNA until 24 h after LPS shock.

Objective To explore the inner feelings of gestational diabetes at their primary use of the real-time blood glucose monitoring. Method Using the qualitative method, 10 gestational diabetic patients monitored their blood glucose using the real-time blood glucose monitoring for the first time and then were interviewed, the acquired results treated with phenomenological analysis. Result The inner feelings of the 10 patients were summarized including 5 themes:worry about the safety of the fetus, fear on alarming at monitoring, different views on four times input of finger blood glucose values, worry about inaccuracy in blood glucose monitoring data, changes in mental conditions and diets. Conclusion Health care workers should provide the gestational diabetic patients with targeted care strategies based on presence of psychological status and knowledge needs so as to improve their compliance, reduce the psychological burden of patients and control their blood glucose level.

Objective To investigate the effect of different doses of lidocaine on expression of the high mobility group box 1 protein (HMGB1) in small intestine in septic rats.Methods Fifty adult male Wistar rats weighing 200-250 g were randomly divided into 5 groups ( n =10 each):sham operation group (group S),sepsis group( group Sep ) and different doses of lidocaine group (group L1～3 ).Group S were not applied cecal ligation and puncture (CLP).Sepsis intestinal damage model was performed in group Sep by CLP.Group L1～3 were given intraperitoneally lidocaine in a dose ofS,10 and 20 mg/kg at immediately,1 and 2 h after CLP,respectively.Isometric normal saline was given intraperitoneally in group S and group Sep.The small intestine tissues were taken at 24 and 48 h after CLP.The small intestine morphology was observed with optical microscope.The expression of the HMGB1 mRNA were examined by PCR and the activity of diamine oxidase (DAO) were determined by ELISA.Results Compared with group S,the expression of HMGB1 mRNA was increased and the activity of DAO decreased in group Sep and groups L1～3 at 24 and 48 h after CLP ( P ＜ 0.05 ).Compared with group Sep,the expression of HMGB1 mRNA was decreased and the activity of DAO increased in a dose-dependent manner in groups L1～3 ( P ＜0.05),The injury of pathology of small intestine was slighter in groups L1～3 than in group Sep.Conclusion Lidocaine can reduce samll intestine injury through inhibiting HMGB1 expression in septic rats by a dose-dependent manner.

In order to understand the intracellular mechanism of preconditioning, we investigated the relationship among activities of extracellular signal-regulated protein kinases (ERKs), the expression of hypoxia-inducible factor -1alpha (HIF-1alpha) and the effect of hypoxic preconditioning (HPC) on cell injury induced by hypoxia-reoxygenation in cultured neonatal rat cardiomyocytes 24 h after brief hypoxia. Cultured cardiomyocytes of neonatal Sprague-Dawley rats were divided into four groups: hypoxia/reoxygenation (H/R), hypoxia preconditioning (HPC), hypoxia preconditioning + mitogen-activated protein kinase (MAPK) inhibitor PD98059 (HPC+PD98059), and control (C). We measured the survival rate and apoptosis rate of cardiomyocytes at 6 or 12 h after hypoxia/reoxygenation, activities of extracellular signal-regulated protein kinases (ERKs), and expression of hypoxia-inducible factor-1alpha (HIF-1alpha). We found that the survival rate of cardiomyocytes in hypoxic preconditioning group increased by 6.08% and 7.91% at 6 and 12 h after hypoxia/reoxygenation (n=6, P<0.05), respectively, and the apoptotic rate decreased by 10.92% and 14.34% (n=6, P<0.05) respectively. Hypoxic preconditioning increased the abundance of phospho-ERK1/2 by 3-folds and expression of HIF-1alpha by 1-fold in whole cell extracts from hypoxic preconditioned cardiomyocytes. PD98059, an inhibitor of the upstream kinase of ERKs, abolished the anti-injury effect, ERKs activation, and expression of HIF-1alpha induced by hypoxic preconditioning. Statistical analysis indicated that there was negative correlation between apoptotic rate and activities of ERKs or expression of HIF-1alpha, and positive correlation between activities of ERKs and expression of HIF-1alpha. It is concluded that hypoxic preconditioning protects cardiomyocytes from hypoxia/reoxygenation-induced injury and that upregulation of HIF-1alpha through ERKs pathway mediates the cardioprotection of hypoxic preconditioning.
Animals ; Animals, Newborn ; Apoptosis ; drug effects ; Cell Hypoxia ; Cell Survival ; Cells, Cultured ; DNA-Binding Proteins ; biosynthesis ; physiology ; Extracellular Signal-Regulated MAP Kinases ; metabolism ; Hypoxia-Inducible Factor 1 ; Hypoxia-Inducible Factor 1, alpha Subunit ; Ischemic Preconditioning, Myocardial ; Myocytes, Cardiac ; cytology ; pathology ; Nuclear Proteins ; biosynthesis ; physiology ; Rats ; Rats, Sprague-Dawley ; Transcription Factors ; biosynthesis ; physiology

OBJECTIVETo investigate the correlation between Chinese medical syndrome type (CMST) and the ID4 gene promoter methylation state in the bone marrow cells of acute myeloid leukemia (AML) patients, and to discuss the correlation between ID4 gene methylation and the occurrence, development of AML.

METHODSThirty-five inpatients or outpatients from the Department of Hematology, the Affiliated Hospital of Shandong University of Traditional Chinese Medicine were recruited as the test group, while 10 healthy volunteers from the health medical center of the Affiliated Hospital, Shandong University of Traditional Chinese Medicine were recruited as the control group. The ID4 gene promoter methylation states were detected using methylation specific polymerase chain reaction (MS-PCR) in the two groups. Inter-group comparison was performed and CMSTs were compared to analyze the correlation between CMSTs and the gene promoter methylation.

RESULTSTwenty-seven AML patients (77.1%) had methylation of ID4 gene, showing statistical difference when compared with the control group (P < 0.01). The ID4 methylation positive rate of ID4 gene promoter methylation was sequenced from low to high as qi and yin deficiency syndrome < inter-accumulation of blood stasis and phlegm syndrome < toxic heat inflaming syndrome, showing statistical difference when compared with the control group (P < 0.05). The peripheral white blood cells and the bone marrow blast cells were higher in ID4 methylation positive patients than in the ID4 methylation negative control patients with statistical difference (P < 0.05).

CONCLUSIONSPatients of inter-accumulation of blood stasis and phlegm syndrome and toxic heat inflaming syndrome were more likely to have ID4 gene methylation. The ID4 methylation positive expression has verified the essence of evil domination in the early AML at the molecular level. It can also reflect the degree of malignancy of AML to some extent.

Adolescent ; Adult ; Aged ; Case-Control Studies ; DNA Methylation ; Female ; Humans ; Inhibitor of Differentiation Proteins ; genetics ; metabolism ; Leukemia, Myeloid, Acute ; diagnosis ; genetics ; metabolism ; Male ; Medicine, Chinese Traditional ; Middle Aged ; Promoter Regions, Genetic ; Young Adult

Objective To evaluate the value of noninvasive monitoring of pulmonary arterial pressure in the children with severe pneumonia and respiratory failure.Methods A prospective study was adopted to investigate 69 patients who suffered from severe pneumonia and respiratory failure in Pediatric Intensive Care Unit in Shanghai Children's Hospital from June 2013 to December 2013 were involved in this study,except for heart disease.The pulmonary arterial pressure (PAP) and cardiac function were monitored by using bedside color doppler ultrasound cardiogram,such as PAP,cardiac index (CI),left ventricle ejection fraction(LEFT),and heart early diastolic filling velocity maximum/heart late diastolic filling velocity maximum (E/A ratio).They were divided into 2 groups according to PAP,one group as pulmonary arterial pressure normal group,the other group as pulmonary arterial hypertension(PAH) group,and the impact of the PAP on the prognosis and mechanical ventilation was assessed.Milrinone[0.5 μg/(kg · min)] were given the patients who were combined with pulmonary hypertension,and the PAP and cardiac function before using Milrinone and 24 h,48 h and 72 h after giving medicine was observed.Results Among 69 cases,40 cases were male and 29 cases were female,age ranging from 2 months to 12 years old,and the weight range was (14.3 ± 8.9) kg.The pediatric critical illness score(PICS) was 70.5 ± 9.6,and the pediatric risk of score m ortality Ⅲ was 13.5 ± 5.0.Among 69 cases,46 cases had pulmonary arterial hypertension,38 cases of them experienced mechanical ventilation,and 9 cases died.Among 23 cases who had no pulmonary arterial hypertension,only 8 cases experienced mechanical ventilation.There was a significant difference in the mechanical ventilation rate and mortality between two groups(x2 =15.78,P ＜0.0l ; x2 =5.18,P ＜ 0.05).The mechanical ventilation time was longer in pulmonary arterial hypertension group (t =3.89,P ＜0.01).PAP was (58.23 ±5.44) mmHg(1 mmHg =0.133 kPa),(49.10 ±4.69) mmHg,(42.53 ±4.54)mmHg and(35.63 ±4.78) mmHg respectively before and after using Milrinone 24 h,48 h and 72 h in 46 cases with pulmonary arterial hypertension,and the pressure decreased significantly after using medicine (F =67.11,P ＜ 0.01).There was no significant difference in CI,LVEF and E/A(all P ＞0.05).However,9 cases of them did not show any response to Milrinone,and in the end they couldn't live without mechanical ventilation,they died.Conclusions Noninvasive pulmonary arterial pressure monitoring could be beneficial in judging patient's condition and assessing prognosis of children with severe pneumonia and respiratory failure,and milrinone could decrease PAP.

Objective To explore the clinical features and diagnostic methods of tuberculosis infection in PICU,and improve the understanding of tuberculosis.Methods We analysed the clinical features and diagnostic methods of severe tubercle bacillus infectious diseases in PICU from Jan 2009 to Dec 2012.Severity of disease was graded by pediatric critical illness score.The diagnosis of the pulmonary tuberculosis was in accord with the diagnostic criteria of paediatric pulmonary tuberculosis established by Chinese Medical Association paediatrics branch.And the diagnosis of tuberculosis meningitis and tuberculosis peritonitis based on the clinical physical examination,laboratory examination and pathologic finding.Results Among 22 cases enrolled in this study,totally 16 cases were pulmonary tuberculosis,6 cases were extrapulmonary tuberculosis,and 3 cases were tuberculosis meningitis.The clinical feature of severe tuberculosis infection in PICU was accompanying with one or multiple organ dysfunction besides tuberculosis infection symptom,among them,respiratory dysfunction occurred in 16 cases,cardiovascular dysfunction was observed in 2 cases,and central nervous system dysfunction was found in 3 cases,even 1 patient experienced cardiovascular system dysfunction,respiratory disorder as well as gastrointestinal system dysfunction simultaneously.Sixteen cases of pulmonary tubercle bacillus infection manifested respiratory failure besides fever,cough,shortness of breath and tuberculosis toxicosis symptom,2 cases of them developed into acute respiratory distress syndrome,8 cases needed mechanical ventilation.Two cases of pericardial effusion presented cardiac tamponade.The level of adenosine deaminase elevated in 12 cases,and the positive result of enzyme-linked immunospot assay for tubercle bacillius was observed in 14 cases.Conclusion It is very important to be aware of that severe tubercle bacillus infection exist in critically ill patients admitted in PICU,measuring the level of adenosine deaminase and taking enzyme-linked immunospot assay for tubercle bacillius test are important accessory examination for tuberculosis diagnosis in children.

Objective To study the changes of P-selectin and E-selectin in pediatric patients with critical illness ,and analyze their relationship with the severity and prognosis of diseases.Methods Forprospective study,42 critically ill patients admitted in pediatric intensive care unit (PICU ) from September,2012 to March,2013 as critically ill group were enrolled,and blood specimens were collected with 24 hours after admission.Another 42 cases blood samples were collected from children's physical examination as control group.The severity of the critically ill patients were evaluated by Pediatric Critical illness Score (PICS)and Pediatric risk of score mortality (PRISM)-III.The levels of serum P-selectin and serum E-selectin were measured by double antibody sandwich enzyme-linked immunoassay (ABC-ELISA). Results P-selectin and E-selectin in control group children and critically ill patients group were (37.23 ± 8.99)ng/mL,(36.24 ±17.82)ng/mL,and (107.24 ±35.53)ng/mL,(114.93 ±40.17)ng/mL, respectively.There were statistical differences between two groups (P=0.000).The levels of P-selectin and E-selectin in acute phase were higher than that of levels in recovery phase in critically ill group (P =0.000).Negative correlation was observed between P-selectin concentration and the PCIS score (r =-0.673,P=0.000),as well as E-selectin (r=-0.548,P=0.000).P-selectin level and E-selectin level based upon PRISMⅢ≥10 group were significantly higher than they in PRISMⅢ <10 group (P=0.003,P=0.014).In critically ill children,the differences in P-selectin,E-selectin were significant higher in death patients (P=0.003;P =0.000).Compared with the non-sepsis illness group,the level of P-selectin and E-selectin in the severe sepsis patients were significantly higher (P =0.04,P =0.025 ). Conclusions The levels of P-selectin and E-selectin are closely related to the severity and prognosis in critically ill children.Measuring the level of P-selectin and E-selectin could be used as a judegment the severity and to understand pathological physiological process.

Objective To investigate the changes of epithelial neutrophil activating peptide-78 (ENA-78) in the serum of patients with critical illness,and to analyze the relationship between the severity and prognosis.Methods Prospective case-control study was performed,and 42 cases of critically ill patients admitted to Pediatric Intensive Care Unit,Children's Hospital Affiliated to Shanghai Jiaotong University from Sep.to Nov.2013 were selected as critically ill group,blood specimens were collected within 24 hours and 7 days after their admission.Another 42 cases of blood samples were collected during physical examinations in this hospital as control group.The severity of critically ill patients were graded by Pediatric Critical Illness Score (PICS) and Pediatric Risk of Score Mortality (PRISM) Ⅲ,and the serum ENA-78 was measured by double antibody sandwich enzyme-linked immunoassay.Results 1.The level of ENA-78 in the control group was (0.44 ± 0.28) ng/L; ENA-78 in acute phase and recovery phase of critically ill group were (2.85 ± 0.89)ng/L and (1.00 ± 0.64)ng/L,respectively,there were statistical differences between control group and critically ill group,acute phase group and recovery phase group (all P =0.000).2.The negative correlation was observed between ENA-78 concentration and PCIS score(r =-0.724,P =0.000).ENA-78 in PRISM Ⅲ ≥ 10 group was significantly higher than that in PRISM Ⅲ＜ 10 group(P =0.000).The ENA-78 between death group and the survival group was significantly different(P =0.000).3.ENA-78 in patients with severe infection was higher than that in the non-infectious cases(P =0.000).4.With the organ dysfunction expanded ENA-78 rose accordingly,and the difference was statistically significant (P =0.000).Conclusions The level of ENA-78 is different in critically ill patients in children.It can provide reference of assessing the severity of disease and predicting prognosis by determing the ENA-78 level.

Objective To study critical hemophagocytic syndrome (HPS) or macrophage activation syndrome (MAS) presented with multiple organ dysfunction syndrome (MODS) in pediatric intensive care unit (PICU),including clinical features and outcomes In order to explore the effect of bedside continuous hemodialysis/hemofiltration (CBP) as adjuvant treatment for severe HPS/MAS.Methods A total of 19 children with HPS/MAS were hospitalized met the diagnostic criteria for HPS from January,2009 to December,2012.Twelve cases were treated with CBP by continuous venin-venin hemodialysis/hemofiltration (CVVHDF) or high-volume hemofiltration (HVHF) following conventional anti-inflammatory therapy.The replacement liquid dose was 50-75 ml/ (kg · h).The organs function were evaluated and laboratory biomarkers including blood 、electrolytes,ferritin changes were measured before and after CBP treatment.Results Ninteen cases of HPS were acute onset and developed to MODS rapidiy after admission to PICU.The main clinical features were the irregular fever or high fever,hepatosplenomegaly and significant liver damage,nervous system dysfunction and disseminated intravascular coagulation (DIC).Eight cases were death and mortality rate was 42.1％,and all death occurred in those aged less than 3 years old.The mortality rate were 25％ (3/12) and 71.4％ (5/7) in CBP group and non-CBP group respectively.After CBP for 6-24 hours,the fever returned to normal range and blood electrolytes improved.The serum ferritin,serum alanine aminotransferase (ALT) and lactate dehydrogenase (LDH) reduced significantly.Serum creatinine (sCr),blood urea nitrogen (BUN) level improved.Four cases with acute respiratory distress syndrome (ARDS) improved and the ventilator parameters were downregulated.Conclusions Our findings indicate that HPS/MAS complicated with MODS is life threatening with high mortality rate.CBP therapy can lower the fever within a short time,correct electrolyte imbalance,stable circulatory function,improve the lung,liver,and brain function.It is suggested that CBP may be the potential effective therapy in severe HPS/MAS with MODS in children.