Objective To introduce the nursing care of severe asthma patients treated by sedatives and muscle relaxants combined with mechanical ventilation. Methods Nurses with more than 3 years of working experiences in respiratory intensive care unit participated in the nursing care,such as ventilation monitoring,sedative effect assessment and artificial airway management. Results All the 18 patients were treated successfully,and the duration of mechanical ventilation ranged from 6 to 192 hours. The indicators of circulation and arterial gas after weaning were improved significantly (P<0.01). Conclusion Proper nursing care and correct sedative effect assessment is an important point to ensure treatment efficacy and reduce complications for severe asthma patients treated by sedatives and muscle relaxants combined with mechanical ventilation.

Objective Inflammatory bowel disease is an important chronic gastrointestinal disease of childhood and adolescence.Intestinal mucosa barrier damage plays an important role in its pathogenesis.This study attempts to use IL-1β stimulating Caco-2 cell monolayer simulates inflammatory intestinal epithelial barrier in vitro,provides the basis for studying the pathogenesis and treatment of IBD.Methods Caco-2 cells were cultivated in vitro until the 21st day to simulate intestinal epithelial cell monolayer barrier.The cells of inflammation group one were disposed with IL-1β for 2 h since the 5th day and detected TEER every other day until the 21st day.The cells of inflammation group two were disposed with IL-1 β at the 18th day for 0,12,24,48,72h, and detected TEER respectively.Normal control group cells were cultured with common medium and detected TEER at the corresponding time point.Results The TEER of Caco-2 cells gradually increased from the 5th to 15th days,reached 600Ω·cm2 in the 15th day of a plateau until to the 21st day.Since the 5th day,the TEER of inflammation group one were all lower than normal group,and still to the 21st day ＜ 500Ω·cm2.Inflammation group two shows the time dependence TEER gradually reduce,peaked at 48 hours,then a slight increase in 72 hours.Conclusion The Caco-2 cells cultured for 2 ～ 3 weeks can form intestinal epithelial monolayer barrier with polarity,then treated with IL-1 β can manufacture inflammatory intestinal epithelial barrier model in vitro.

Objective To evaluate the efficacy and safety of bicyclol tablet on hepatic lesions caused by Epstein-Barr virus( EBV) infection in children.Methods A single-center controlled retrospective study was conducted in 121 children with hepatic lesions caused by EBV infection for evaluating safety,tolerability, and efficacy of treatment with bicyclol tablets or Glycyrrhizin capsules.Childer n in bicyclol group ( n=63 ) were treatedw ith bicyclol at blets and cotn rol group ( n =58 ) were treated with Glycyrrhizin capsules.The course of the treatment were both 8 weeeks for two groups.The level of the EBV load pretreatment and plas-ma aminotransferase,blood routine,urine routine pretreatment and 1 week,4 weeks and 8 weeks after treat-mentw ere analyzed er trospectively.Rse ults (1) The pal smaA LT level sigin ficantly decreased in theb icy-clol group compared with that in the contro l group(P<0.01), se pecai lly the levle after 8 weeks treatm ent. (2) Bicycol l was more effce tive in the bicyclol group than Glycyrrhizin capslu se in the control group( P<0.01).(3) Both grousp had no significantlya dvesr e events.Conclusion Bicyclol tablet can derc ease plas-ma aminotransferase level,espce ially ALT,inc hildren caused by EBV infection with better efficiency and safety.

Objective Intestinal epithelial barrier damage is closely related to a variety of gastrointestinal disease,how to maintain its function effectively is the key to treat all these diseases.This research attempts to explore the protective effect and its mechanism of toll-like receptor 2 (Toll-like receptor,TLR2)on permeability of intestinal epithelial barrier by experiments in vitro,to lay a foundation for new treatment methods.Methods We cultured non-transfected Caco-2 cells,TLR2-deficiency Caco-2 cells,TLR2-overexpressed Caco-2 cells in normal control group until the 21 st d,then tested transepithelial electrical resistance(TEER) which reacts the permeability of epithelial barrier.We cultured 3 types of cells in inflammation group until the 19th d treated with 10 ng/ml IL-1 beta for 48 h,then tested TEER values at the 21st d.We treated 3 types of cells in inhibition group with PI3K/Akt pathway inhibitor for 1h befor IL-1 beta,then tested TEER values at the 21st d.Results TEER value of TLR2-deficiency Caco-2 cell monolayer significantly reduced (P ＜ 0.01),whereas TEER value of TLR2-overexpressed Caco-2 monolayer raised,but without statistically significant.TLR2 can prevent IL-1 beta caused TEER decreasing (P ＜ 0.01),but the effect disappeared after given PI3K/Akt pathway inhibitor.Conclusion TLR2 can regulate the permeability of intestinal epithelial barrier.In addition,TLR2 can protect permeability increasing caused by inflammation,this effect mediated by PI3 K/Akt pathway.

Objective Gastrointestinal dysfunction is closely correlated with the impairment of intestinal barrier caused by serious infection.We focused on the role of platelet activating factor(PAF)in the intestinal impairment caused by endotoxemia by studying on the morphology of intestinal epithelial ceils and diamine oxidase (DAO)levels with the application of Ginkgolide B(PAF receptor antagonist).Method Eighteen-day-old Wistar rats were randomized into lipopolysaccharide(LPS)(5 mg/kg),PAF receptor antagonist(pretreatment and treatment)and normal saline(Control)groups(n=8 at each time point).Ginkgolide B(PAF receptor antagonist BN52021)5 mg/kg was administered 30 minutes before(in pretreatment group)and after LPS injection(in treatment group).The ileum specimens(n=8)were harvested at 1.5,3,6,24,48 and 72 hours after LPS or NS injection.The ultrastructures of intestinal epithelial cells were studied by transmission electron microscopy (TEM)and with hematoxylin and erosin staining.The contents of DAO in ileum tissue and plasma were measured respectively with spectrophotometer.According to data Normality and Variance equality,ANOVA analysis and LSD (least significant difference)-t test were used for multiple group difference.The whole test Was performed in the animal laboratory,pathological laboratory,biochemical laboratory of our hospital and electron microscopy laboratory of Liao-ning University of Traditionary Chinese Medicine.Results Histologic examination of intestinal injury in LPS group showed the edema of intestinal villi,the capillary congestion in lamina propria,the dilation of interstitial lymphatic vessel.and the polymorphonuclear infiltration in enteric cavity in LPS group at 1.5,3,6,24 hours.The edema of the intestinal villi were shown in antagonist group.Ultrastructural study showed microvilli and tight junctions were intact in the control group.The tight junctions enlarged and the microvilli were thin,rare or disrupted in the experimental group.The pathological changes in PAF antagonist group were slightly lighter than that in the LPS group.The DAO content in the ileum tissue was obviously decreased in the LPS group compared with that in the control group.It reached to a nadir at 6 hrs[from(0.172±0.004)U/mg to(0.096±0.010)U/mg,F=13.372,P＜0.01).The DAO content in plasma was obviously higher in the LPS group than that in the control group.The patterns of DAO changes in the PAF antagonist group were as the same as that in the LPS group at each time point.Conclusions PAF may play a certain role in the injury of intestinal barrier in endotoxemia.Preventive and remedial administration of Ginkgolide B may relieve intestinal injury.The activity changes of DAO in plasma synchronized with that in ileum tissue.It may be deduced that the alterations of DAO in plasma may indicate the destruction of intestinal mucosa in the early stage sensitively.

Objective To evaluate the protection of combined clostridium butyricum and bifidobac-terium living powders for antibiotic-associated diarrhea ( AAD ) with all kinds of infections in hospitalized children,and to compare the therapeutic effect with saccharomyces boulardii. Methods This study was a prospective,randomized case-control clinical trial which collected the data of the hospitalized children with all kinds of infections in Pediactric Department of Shengjing Hospital of China Medical University between May 2011 to May 2012. A total of 552 cases were enrolled and 480 cases completed the study. A total of 240 chil-dren were in experimental group,80 cases received combined clostridium butyricum and bifidobacterium liv-ing powders 840 mg per time,twice a day and the other 160 cases received saccharomyces boulardii 250 mg per time,twice a day,for one week; the control group took none of probiltics. Two groups received routine antibiotic therapy. Everyday′s defecate frequency was recorded, the traits of excrement according to bristol stool assessment scale were evaluated,the incidence of diarrhea and drug related adverse reactions were coun-ted. Results During the studied 7 days,the AAD incidence was 4. 2%(10/240) in experimental group and 20. 4%(49/240) in control group,there was significant difference between two groups. The risk of AAD in experimental group decreased 58. 5%. Compared to saccharomyces boulardii,combined clostridium butyricum and bifidobacterium living powders decreased 38. 2% (RR=0. 728, 95%CI 0. 257~0. 784, P=0. 009). Compared to control group,the average defecate frequency decreased in experimental group,diarrhea duration contracted,there was statistic difference between two groups ( P<0. 01 ) . No drug related adverse reactions happened during the trial. Conclusion Both combined clostridium butyricum and bifidobacterium living powders and saccharomyces boulardii could effectively reduce the risk of AAD in hospitalized children with bacterial infection,relieve diarrhea symptoms,short the duration of diarrhea,and did not find the adverse reac-tions. Combined clostridium butyricum and bifidobacterium living powders and saccharomyces boulardii had the same protective effect for AAD of northern China children.

Objective To establish and evaluate intestinal epithelial barrier model using Caco-2 cell so as to play a foundation for next study of barrier permeability.Methods Caco-2 cells were cultured in vitro then seeded into Transwell cell culture inserts.The permeability of the intestinal epithelial barrier was detected by transepithelial electrical resistance(TEER)and lucifer yellow flux,and verified by transmission electron micro-scope.Different concentrations of PAF(0,50,100,and 200 nmol /L)were exposed for 24 hours to Caco-2 mono-layer when cultured 21 days.The tight junction was observed under transmission electron microscope.Assess-ment of ZO-1 protein localization and expression were detected by immunofluorescence and Western blot analy-sis.Results Cultured Caco-2 cell confluencd as monolayer with time passed.From 5th day,TEER increased, then reached 600Ω?cm2 at 15th day and lasted to 21 st day,there was little flux of lucifer yellow,transmission e-lectron microscopy also found cells differentiated better,had well-arranged villi and polarity alined as monolayer, forming completed tight junction which was the marker of intestinal epithelial barrier model in vitro.TEER de-creased and lucifer yellow flux increased in cells exposed to PAF.The permeability reached the peak when ex-posed to 100 nmol /L PAF(P <0.01 ),tight junction disrupted,ZO-1 protein expression downregulated,abnor-mal localization and distribution was assessed by immunofluorescence staining.Conclusion Cultured Caco-2 cells for 2-3w can be used to study intestinal epithelial barrier as a model in vitro.PAF increased intestinal epi-thelial permeability,which would correlate to the decreased protein expression and abnormal distribution of ZO-1.

ObjectiveTo investigate the significances of clinical diagnostic approaches to biliary atresia (BA) and intrahepatic cholestasis (IHC) in children, and to improve clinicians′ understanding of BA and reduce the rates of missed diagnosis and false diagnosis. MethodsA total of 133 children diagnosed with cholestasis with persistent jaundice admitted to our department from July 2011 to June 2014 were divided into IHC group with 111 patients and BA group with 22 patients. The general clinical trial data were reviewed and analysed and the significances of clinical manifestations, laboratory examination, and imaging features for differential diagnosis of BA and IHC were evaluated. Comparison of continuous data between the two groups was made by t test and comparison of categorical data between the two groups was made by chi-square test. When the sample characteristics for chi-square test were not suitable, the comparison was made by Fisher′s test. ResultsSignificant differences in clinical manifestations of kaolin stools and enlarged and hardened liver and spleen were observed between the two groups (P＜0.01). Total bilirubin (TB), direct bilirubin (DB), gamma GGT (γ-GT), alanine aminotransferase (ALT), and aspartate aminotransferase (AST) levels in BA group were significantly higher than those in IHC group (P＜0.01). The specificity, positive predictive value, and positive likelihood ratio of γ-GT were the highest among them, while TB had the highest sensitivity, the highest negative predictive value, and the lowest negative likelihood ratio. The sensitivity and negative predictive value of TB, DB, and γ-GT in the parallel experiment reached 100%. The specificity and positive predictive value of them in the serial experiment were 98% and 88.9%, respectively. There were significant differences in the hepatic portal fibrous mass and gallbladder hypokinesia detected with ultrasonography of the liver, gallbladder, and spleen and in magnetic resonance cholangiopancreatography (MRCP) features between the two groups (P＜001). The specificity and positive predictive values of them in serial experiment reached 100%. ConclusionKaolin stools, enlarged and hardened liver and spleen, TB, DB, γ-GT, ultrasonography, and MRCP of the liver and gallbladder are important indices to distinguish between BA and IHC and a combined analysis of them can improve the diagnostic accuracy.

Progressive familial intrahepatic cholestasis (PFIC) is a rare heterogenous groups of autosomal recessive monogenic disease, resulting in abnormal bile acid formation and secretion.According to the type of gene mutation, PFIC was classified as PFIC 1-6 and more new mutated genes have been identified, such as ABCC12, VPS33B, et al, making diagnosis and treatment more accurate.PFIC is characterized by progressive jaundice and pruritus, growth and development disorders, and fat-soluble vitamin deficiency.Except for PFIC3, the level of γ-glutamyl transpeptidase in serum of PFIC is normal, which is an important clue for clinical diagnosis.However, each type of PFIC has its own characteristics such as the age of onset, disease severity, extrahepatic symptoms, and prognosis.With the progress of the disease, it eventually develops into fibrosis, cirrhosis and liver failure.Treatment includes nutrition, medication, partial external/internal biliary diversion, and liver transplantation.Recent studies on transporters and gene therapy have brought new hope to patients with PFIC.

Matrix metalloproteinases(MMPs)are a large family of zinc-dependent endopeptidases, which are mainly synthesized by connective tissues, it can degrade the extracellular matrix(ECM)and basement membrane, affect the regeneration and reconstruction of normal tissues, and participate in the pathological process of malignant tumors.Matrix metalloproteinase-7(MMP-7)is the smallest member of the metalloproteinase family.It is expressed in many tissues of the body, such as thyroid, breast, lung, digestive tract, reproductive system and hepatobiliary system.In recent years, the expression of MMP-7 in hepatobiliary diseases has attracted more and more attention.MMP-7 is not only involved in the growth, metastasis and invasion process of hepatobiliary malignant tumors, but also highly expressed in liver fibrosis, biliary atresia and other diseases.This paper reviews the expression of MMP-7 in the above diseases.