Objective Retinoic acid receptor(RAR?) selective antagonist,Ro41-5253(Ro) was used in the counteract test,to confirm the role of RAR? in enhancement of IgM synthesis in cord blood lymphocytes(CBLs) by retinoic acid(RA).Methods CBLs were cultured in vitro and stimulated with or without RAR? agonist RA and(or) antagonist Ro.The cells were harvested at 24 hours and 48 hours of culture time to measure the levels of gene expression of RARa,IL-4 and IL-6.ELISA was used to measure the concentration of IgM in the supernatant of 5 days culture.Results Ro could inhibit the promotion of RA on IgM synthesis in CBLs.RA could up-(regulate) RAR? gene expression,which could be restrained by Ro.Ro also could counteract the up-regulation of RA on IL-4 and IL-6 gene expression.Conclusion The effect of RA on IgM synthesis in CBLs is modulated by RAR?.

ObjectiveTo explore the therapeutic effect of continuous veno-venous hemofiltration (CVVH) on the treatment of severe acute pancreatitis (SAP).MethodsAll data about forty-five patients with SAP admitted to the intensive care unit (ICU) from June 2005 through June 2010 were reviewed.These 45 patients were randomly (random number ) divided into routine treatment group (n =22 )and comprehensive treatment group ( n =23 ).In control group,patients were rapidly given with a suffficient liquid support,vasoactive drug to increase organ perfusion,trypsin secretion inhibitor,broad-spectrum antibiotics,enteral nutrition with intestine membrane protective agent in early stage.In the comprehensive treatment group,patients received CVVH integrated with routine treatment.On admission and 72 h posttreatment,the scores of acute physiology and chronic health evaluation Ⅱ ( APACHE Ⅱ ) and multiple organ dysfunction syndrome (MODS),and the results of standard bettery of biochemistry tests indcluding blood urea nitrogen (BUN),serum cratinine (Scr),total bilirubin (TBIL),alanine aminotransferase (ALT),amylase (AMS),C-reactive protein (CRP),TNF-α,IL-6,IL-8 were observed.Time of mechanical ventilation support,length of ICU stay,and survival rate were compared between two groups.ResultsOn admission between the two groups,no statistical significance was seen in the APACHE Ⅱ and MODS score,BUN,Scr,TBIL,ALT,AMS,CRP,TNF-α,IL-6,IL-8 (P ＞ 0.05).But APACHE Ⅱ and MODS score were decreased significantly in comprehensive treatment group than in the routine treatment group,as well as the the level of BUN,Scr,TBIL,ALT,AMS,TNF-α,IL-6,IL-8 and CRP after 72h post-treatment (P＜0.05 ).In routine treatment group and comprehensive treatment group,the time of respirator intervention and length of stay in ICU were (7.6±3.4) d vs.(11.5±4.7) d,(12.3±7.8) dvs.(17.6±9.2) d respectively,the statistical significance was shown ( P ＜ 0.05 ).Compared to the comprehensive treatment group ( 86.96％ ),the survival rate ( 59.09％ ) were lower in routine treatment group ( P ＜ 0.05 ).ConclusionsCVVH combined with routine treatment,which can remove inflammatory agents and toxins,maintain homoeostasis,and improve oxygenation,is effective in treatment of SAP and can improve patient survival rate.

Objective:To determine the di-allelic polymorphisms of TNF gene and their association with Helicobacter Pylori(H.pylori) infection and gastroduodenal diseases in Chinese population with Han nationality.Methods:Two hundreds and ten patients with gastroduodenal diseases(73 chronic gastritis,78 duodenal ulcer and 59 noncardia gastric cancer) and 264 healthy controls were genotyped by the PCR-RFLP method for TNF-? 308,lymphotoxin-?(LT-?) Nco Ⅰ and AspH Ⅰ gene polymorphisms.H.pylori infection status was determined by a validated serological test.Results:H.pylori infection was detected in 90.5% of 210 patients and 62.1% of 264 healthy controls(P

Objective:To investigate the psychological state of the patients from Wenchuan Earthquake treated in some local and military hospitals.Methods:We consulted the general data and pathogenetic condition of 116 patients from the earthquake-stricken area of Sichuan,studied their mental health using the Hamilton Anxiety Scale(HAMA) and investigated their psychological changes during the rescue and treatment with self-designed questionnaires.Results:The earthquake patients manifested obvious anxiety in the initial stage and their mental health was gradually improved towards the end of the rescue process.Conclusion:Wenchuan Earthquake caused great psychological trauma in the earthquake patients,for whom timely psychological intervention is of utmost importance.

Objective To observe the effect of dexmedetomidine on plasma SDF-1 level in in hepatic portal occlusion operation.Methods Fifty patients with live cancer undergoing elective partial hepatectomy were selected,no gender limitation,aged 42 to 71,body mass index(BMI) 18.5 ～ 26.0 kg/m2,ASA grade Ⅱ or Ⅲ.The patients were randomly divided into 2 groups(n=25):control group and dexmedetomidine group.The dexmedetomidine group was performed the pump injection of dexmedetomidine 1 μg/kg at 15 min before induction of anesthesia.After induction the rate was changed to 0.4μg · kg-1 · h-1 until 15 min before the end of operation;the control group adopted the same method for conducting continuous intraverous infusion of the same capaci ty of 0.9％ sodium chloride.The peripheral venous blood was collected in 2 groups at preoperative 1 h (T0),postoperative 1 h (T1),postoperative 1 d (T2),postoperative 3 d(T3).The plasma SDF-1 level was detected by using enzyme-linked immunosorbent assay(ELISA).Results There was no statistically significant difference in liver resection range,blood loss,first porta hepatis vessel occlusion time,anesthesia time and plasma SDF-1 level before surgery between the two groups (P＞0.05).Compared with pre-operation,plasma SDF-11evel at T1,T2,T3 time point was significantly increased (P＜0.05).The plasma SDF-1 level at T1,T2,T3 time point in the dexmedetomidine group was lower than that in the control group(P＜0.05).Conclusion SDF-1 expression is significantly increased during perioperative period in the patients with hepatic portal occlusion operation,and intraoperative continuous dexmedetomidine can significantly reduce the SDF-1 level,which inhibits the chemotaxis and accumulation of inflammatory ceils to some extent.

Objective To evaluate the changes in the expression of annexin A2 (ANXA2) in lung tissues in rats with hepato-pulmonary syndrome.Methods Healthy 3-4-month-old Sprague-Dawley rats of both sexes,weighing 220-250 g,were randomly divided into 3 groups:control group (group C,n =10),sham operation group (group S,n =10) and hepatopulmonary syndrome group (group HPS,n =20).The rats were anesthetized with intraperitoneal 3％ pentobarbital sodium 0.1 ml/100 g.Hepatopulmonary syndrome was produced by chronic ligation of the common bile duct.After 5 weeks,the rats were sacrificed and lungs were removed for determination of ANXA2 and smooth muscle actin α (SM-α-actin) mRNA and protein expression in lung tissues by RT-PCR and Western blot,respectively.Results There was no significant difference in the expression of ANXA2 and SM-α-actin mRNA and protein between groups C and S (P ＞ 0.05).The expression of ANXA2 and SM-α-actin mRNA and protein was significantly higher in group HPS than in groups C and S (P ＜ 0.05).Conclusion The expression of ANXA2 in lung tissues is up-regulated in rats with hepato-pulmonary syndrome.

Objective To investigate the role of serine threonine protein kinase-1 (Akt1) and Smad3 in lung tissues in development of hepatopulmonary syndrome in rats.Methods Seventy-two healthy male SpragueDawley rats,aged 3-4 months,weighing 200-250 g,were randomly divided into 3 groups (n=24 each):control group (C group),sham operation group (S group) and common bile duct ligation (CBDL) group.The rats were anesthetized with 3％ pentobarbital sodium 45 mg/kg.In group CBDL,laparotomy was performed,the common bile duct was ligated and then the abdomen was closed,while the common bile duct was only exposed,but not ligated and then the abdomen was closed in group S.At 1st,3rd and 5th weeks (T1-3),8 rats were chosen randomly in each group and blood samples were obtained from the abdominal aorta for blood gas analysis.The rats were then sacrificed and lungs were isolated to detect the expression of Aktl and Smad3 mRNA and protein in lung tissues (by RT-PCR and Western blot).The lung tissues were sliced and stained with hematoxylin eosin for examination of the pathological changes of pulmonary capillaries.Results Compared with C and S groups,the expression of Akt1 and Smad3 mRNA and protein in lung tissues was significantly up-regulated at T2,3,and alveolar-arterial oxygen tension difference was increased at T3 in CBDL group (P ＜ 0.05).The pulmonary capillary was obviously dilated at T3 in CBDL group.Conclusion The expression of Akt1 and Smad3 in lung tissues is up-regulated,which may be one of the mechanisms of development of hepatopulmonary syndrome in rats.

Objective To establish a method for culture of rat pulmonary capillary pericytes.Methods Six male Sprague-Dawley rats,aged 6-7 weeks,weighing 200-220 g,were anesthetized and the chest was opened.The pulmonary capillary was isolated by type Ⅰ collagenase digestion and micropore filtration and cultured in highglucose DMEM/F12 1∶1 containing 10％ fetal bovine serum and 0.5％ mixture of penicillin and streptomycin.The morphology and growth of cells were observed with inverted phase contrast microscope.The positive cells of αsmooth muscle actin (α-SMA),desmin,neuron-glial antigen 2 (NG2),cluster of differentiation 31 (CD31) were counted by immunofluorescence.The percentage of positive cells was calculated.Results The microscopic examination showed cells of shuttle shape or star shape,mononuclear cells,binuclear cells occasionally,oval nucleus,rich cell plasma,growth in the shape of vortex or fence,and no contact inhibition.The percentage of positive cells ofα-SMA,desmin,NG2,and CD31 was (99.0± 1.2)％,(96.0±2.1)％,(99.0±0.7)％ and0,respectively.Conclusion The culture method for rat pulmonary capillary pericytes is successfully established.

Objective To evaluate the effect of bilirubin on proliferation of pulmonary microvascular endothelial cells (PMVECs) of rats.Methods Primary PMVECs harvested from 10 healthy male Sprague-Dawleyrats,weighing 120-150 g,aged 2-3 months,were cultured,purified and identified.PMVECs were seeded in low-glucose DMEM culture medium or 96-well culture plates,and divided into 5 groups according to the random number table:control group (group C) and different concentrations of bilirubin groups (B1-4 groups) with 30 wells or48 flasks in each group.In group C,low-glucose DMEM 1 ml was added.In B1-4 groups,5,10,20 and 50 μmol/L bilirubin 1 ml were added,respectively.At 24,48 and 72 h of incubation,proliferation of PMVECs was measured using CCK-8 assay and 3 H-TdR incorporation assay,Akt1 mRNA and Smad3 mRNA expression was measured by RT-PCR,and phosphor-Akt1 (p-Akt1) protein and Smad3 protein expression was detected using Western blot.Results Compared with group C,no significant changes were found in each parameter mentioned above at each time point in B1 and B2 groups,the proliferation of PMVECs was weakened,and Akt1 mRNA,p-Akt1 protein and Smad3 protein and mRNA expression was down-regulated at 48 h of incubation in B3 group,and the proliferation of PMVECs was weakened,and the expression of Akt1 mRNA,p-Akt1 protein and Smad3 mRNA and protein was down-regulated at 48 and 72 h of incubation in B4 group.Compared with group B3,the proliferation of PMVECs was weakened,and the expression of Akt1 mRNA,p-Akt1 protein and Smad3 mRNA and protein was down-regulated at 72 h of incubation in B4 group.Conclusion High concentration of bilirubin can inhibit proliferation of PMVECs and down-regulated expression of Akt1 and Smad3 is involved in the mechanism.

Objective To investigate the effect and mechanism of soluble epoxide hydrolase inhibitor (sEHI) for NF-κB pathway and cell circle arrest of tubular epithelial cell in unilateral ureteral obstruction (UUO) mice model.Methods Thirty-two healthy C57BL/6 male mice performed UUO surgery to induce renal interstitial fibrosis.Animals were randomly divided into 4 groups:sham group (n=8),sEHI (1 mg· kg-1·d-1) group (n=8),UUO group (n=8) and UUO+sEHI (1 mg· kg-1· d-1) group (n=8).Daily sEHI [1-(1-methylsulfonyl-piperidin-4-yl)-3-(4-trifluoromethoxy-phenyl)-urea,TUPS] or 2％ DMSO was applied to mice by oral gavage from day 1 to day 14 after surgery.All mice were sacrificed at day 14 and kidneys were harvested for further analysis.The changes of renal tissue morphology and pathology were observed by Hematoxylin and eosin (HE) and sirius red staining.The expressions of sEH,nuclear factor κB p65 (NF-κB p65) and IκB were measured by Western blotting.The expressions of TNF-α,IL-1β,MCP-1,IL-6,TGF-β,CTGF,collagen-Ⅳ and α-SMA were analyzed by real-time PCR.Immunofluorescence staining of phospho-histone H3 (p-HH3) and Ki67 was performed to determine the stage of cell cycle G2/M arrest.Results The expression and activity of sEH increased in UUO group (P ＜ 0.05).Administration of sEHI inhibited activity of sEH and infiltration of inflammatory cell in tubular interstitial,as well as attenuated tubular damage and tubular interstitial fibrosis.Western blotting analysis revealed administration of sEHI inhibited up-regulated NF-κB p65 and down-regulated IκB in UUO group (P ＜ 0.05).Real-time PCR demonstrated that administration of sEHI obviously decreased the mRNA expression of cytokines and fibrosis markers,including of TNF-α,IL-1 β,MCP-1,IL-6,TGF-β,CTGF,Collagen-Ⅳ,α-SMA (P ＜ 0.05).Immunofluorescence staining showed that there were much more p-HH3 and Ki67 double positive nuclear tubular epithelial cells and interstitial cells in UUO group,compared with Sham group (P ＜ 0.05).Administration of sEHI reduced the number of double positive nuclear cell only in tubular epithelial cells (P ＜ 0.05),but not in interstitial cells.Conclusions In UUO tubular interstitial fibrosis model,sEHI inhibits the activation of NF-κB pathway by down-regulating p65 and up-regulating IκB and ameliorates the infiltration of inflammatory cells.In addition,sEHI plays anti-fibrosis effect by moderating cell cycle G2/M arrest and reducing the excrete of pro-fibrosis factors of tubular epithelial cells.