Objective To explore the value of serum level of interleukin 6 (IL-6),tumor necrosis factor alpha (TNF-α) and high mobility group box-1 protein( HMGB1) in early assessment the severity and prognosis of acute pancreatitis (AP). Methods Thirty-three severe acute pancreatitis (SAP) patients and 38 mild acute pancreatitis (MAP) patients were selected as study objects;and 51healthy individuals were set as control group. Serum IL-6,TNF-α and HMGB1 concentrations were determined by enzyme-linked immunosorbent assay (ELISA),the association of them and the scores of Ranson,APACHE Ⅱ . Balthazar CT,serum biochemical parameters and prognosis was analyzed.Results The serum IL-6 levels of SAP group,MAP group and healthy control group were (553. 72±175.76) pg/ml,(265. 73±179. 95) pg/ml and (16. 43±3. 32) pg/ml;and there were statistical significance of these three groups (all P＜0. 01). There was no significant difference of TNF-α in the three groups (all P＞0. 05). The serum HMGB1 levels of SAP group,MAP group and healthy control group were (11. 48±6. 94)μg/L,(6. 13±5. 80)μg/L and (1. 82±0. 64)μg/L respectively,and there were statistical significant of these three groups (all P＜0. 05). The correlation coefficient of serum HMGB1 with IL-6 and TNF-α were 0. 896 and 0. 724 (P＜0. 01) respectively. The IL-6 level was positively correlated with the scores of Ranson,APACHE Ⅱ and Balthazar CT. The TNF-αconcentration was positively correlated with APACHE Ⅱ score,and the HMGB1 concentration positively correlated with scores of Ranson and Balthazar CT. The levels of IL-6,TNF-α and HMGB1were all positively correlated with the serum creatinine concentration. The IL-6 levels of patients with local and/or systemic complications were significantly higher than those without complications.Conclusion The serum levels of IL-6,TNF-α and HMGB1 are significantly correlated with the severity of pancreatitis,all of them take part in the development of acute renal insufficiency. The high level of serum IL-6 significantly correlated with complications.

Objective To quantify the concentration of peripheral blood plasma neutrophil gelatinase-associated lipocalin (NGAL) in patients with acute pancreatitis (AP) and to explore its value in assessment of the severity of AP.Methods From June 2011 to March 2012,83 patients with AP were selected,among those 43 cases were mild acute pancreatitis (MAP) and 40 were severe acute pancreatitis (SAP).The control group included 30 healthy individuals.The peripheral blood of patients with AP and healthy controls was collected,and plasma was isolated after centrifuged.The concentration of NGAL in plasma was detected by enzyme-linked immunosorbent assay (ELISA).The t-test was performed for comparison between groups.The correlation between the concentration of NGAL in plasma and clinical parameters of AP was analyzed by Spearman rank order correlation analysis.The diagnosis value of the concentration of NGAL in SAP was evaluated by receiver operating characteristic (ROC) curve and area under the curve (AUC).Results The concentration of plasma NGAL in AP group ((10.30± 5.97)nmol/L) was higher than that in healthy control group ((1.94±1.35) nmol/L) and the difference was statistically significant (t=11.924,P＜0.01).The concentration of plasma NGAL in SAP group ((14.61 ±5.28) nmol/L) was higher than that in MAP group ((6.27±-3.09) nmol/L) and healthy control group,the differences was statistically significant (t=8.677 and 14.539,both P＜0.01).The concentration of plasma NGAL of AP patients was positively correlated with acute physiology and chronic health evaluation-Ⅱ score,Ranson score,bedside index for severity in acute pancreatitis score,computed tomography (CT) severity index,C-reactive protein,white blood cells and the days of hospitalization (r=0.651,0.556,0.514,0.620,0.320,0.458 and 0.346,all P＜0.05).The area under the ROC curve of plasma NGAL concentration in diagnosis of SAP was 0.926 (95％CI:0,870-0.983).The cutoff value of plasma NGAL level in diagnosis of SAP was 8.44 nmol/L.The sensitivity and specificity was 87.5 ％ and 88.9％,respectively.Conclusions Plasma NGAL level is correlated with the severity of patients with AP.NGAL may be one of the markers for the early diagnosis of SAP.

Objective To investigate the antitumour effects of transfected gut-enriched Krüppellike factor(GKLF) on human gastric carcinoma cell line SGC-7901 in vitro and in vivo. Methods The expression of GKLF mRNA and protein in human gastric carcinoma cell line SGC-7901 were detected before and after transfection by real-time fluorescence quantitative PCR and Western blot,respectively. Proliferation and invasion in SGC-7901 were measured respectively by MTT assay, flow cytometry, colony formation assay and cell invasion assay after transfected with GKLF. The growth of xenograft was observed, the microvessel density(MVD) of xenograft tissue was determined by immunohistochemistry. Results The GKLF mRNA and protein in SGC-7901 were overexpressed after transfected with GKLF(P＜0.05). The proliferative speed of SGC7901-pcDNA3.1-GKLF group was markedly lower than that of SGC-7901 and SGC7901-pcDNA3.1 groups (P＜0.05). Transfected with GKLF caused part of the G0/G1 arrest, decreased clone formation rate and the invasion ability (P＜0.05). The growth speed of xenograft in SGC7901-pcDNA3.1-GKLF group was lower, the weight and MVD of xenograft tissue in SGC7901-pcDNA3. 1-GKLF group were less (P＜ 0. 05).Conclusion Transfected with GKLF maysuppress proliferation and invasion in human gastric carcinoma cell line SGC-7901, inhibit the growth and the angiogenesis of xenograft in nude mice.

Objective To determine the inhibitory effect and mechanism of exogenous carbon monoxide against excessive neutrophil infiltration in liver and lung tissues during sepsis.Methods Thirty-two mice were subjected to sham operation (sham group),cecal ligation and perforation (CLP) group,CLP with 8 mg/kg of exogenous carbon monoxide releasing molecule Ⅱ (CORM-2) (CORM-2 group),and CLP with 8 mg/kg of inactive variants of CORM-2 (iCORM-2) (iCORM-2 group) according to the random number table,with 8 mice per group.Liver and lung tissues were collected at 24 hours after surgery to examine the pathologic changes,myeloperoxidase (MPO) activity and malonaldehyde (MDA) content.Another 60 mice were enrolled into the same 4 groups with 15 mice per group and were tested for 72-hour survival rate.Bone marrow neutrophils were isolated and divided into normal control group,1 μg/ml lipopolysaccharide (LPS) group,1 μg/ml LPS plus 10 μmol/L CORM-2 group (low dose group),1 μg/ml LPS plus 50 μmol/L CORM-2 group (high dose group),1 μg/ml LPS plus 50 μmol/L iCORM-2 group (iCORM-2 group).Under the agarose chemotaxis,qPCR and immunofluorescence detection of formyl peptide receptor 1 (FPR1) were performed.Results CLP group presented enhanced activity of MPO [liver:(9.1 ± 1.1) U/g,lung:(16.3 ± 2.8) U/g],increased MDA content [liver:(76.5 ±11.3) nmol/mg,lung:(32.4 ± 10.3) nmol/mg] and 72-hour survival rate of 20％ as compared with the sham group (all P ＜ 0.05).CORM-2 group showed inhibited activity of MPO [liver:(5.2 ± 0.8) U/g,lung:(7.5 ± 2.4) U/g],increased MDA content [liver:(46.7 ± 6.1) nmol/mg,lung:(23.8 ±7.3) nmol/mg] and 72-hour survival rate of 67％ as compared with the sham group (all P ＜ 0.05).LPS enhanced neutrophil migration (61.3 ± 7.1) (P ＜ 0.05) and expression of FPR1 which was enriched in the membrane.Meanwhile,neutrophil migration was significantly inhibited in a dose-dependent of CORM-2 (low dose group:43.3 ±6.1,high-dose group:23.3 ±5.9) (P＜0.05).Conclusions Exogenous carbon monoxide is effective to inhibit the excessive neutrophil infiltration,attenuate oxidative stress or pathological injury,and improve the survival from sepsis.The mechanism is associated with the down-regulation of FPR1,inhibition of FPR1 enrichment in the membrane,and decreased neutrophil migration.

ObjectiveTo study the role of Kukoamine B (KB) in inhibiting the inflammatory response of small intestine in septic mice and its molecular mechanisms.Methods Twenty-four male ICR mice were randomly divided into control group, model group, and KB intervention group (each,n= 8). Sepsis model was reproduced by intra-peritoneal injection of 20 mg/kg lipopolysaccharide (LPS), while equivalent normal saline was given in control group, and 20μg/kg KB was injected through caudal vein 4 hours after LPS challenge in KB intervention group. The blood/tissue samples (jejunum and ileum) were harvested 8 hours after LPS injection. The levels of plasma LPS, tumor necrosis factor-α (TNF-α) and interleukin-1β (IL-1β) were measured. The pathological changes in small intestine tissues were observed under light microscope, while the levels of inflammatory cytokines TNF-α and IL-1β in the tissue homogenates (jejunum and ileum) were assessed by enzyme linked immunosorbent assay (ELISA). The activity of myeloperoxidase (MPO) was measured by colorimetry. The expression of intercellular adhesion molecule-1 (ICAM-1) was determined by immunohistochemistry. The expressions of inducible nitric oxide synthase (iNOS) mRNA was assayed by reverse transcription-polymerase chain reaction (RT-PCR). The activation of nuclear factor-κΒ (NF-κΒ) was determined by Western Blot.Results The mice in model group were found to have an increase in microvascular permeability, interstitial edema, and infiltration of white blood cells, and the levels of LPS, TNF-α and IL-1β in their plasma, with an increase in concentrations of TNF-α and IL-1β, activity of MPO, positive expression of ICAM-1, expression of iNOS mRNA and NF-κB protein in small intestine (jejunum and ileum). Compared with model group, in mice with KB intervention, microvascular permeability, interstitial edema, and infiltration of white blood cells were reduced significantly, while the levels of LPS, TNF-α and IL-1β in plasma, the concentration of TNF-α and IL-1β, the activity of MPO, the positive expression of ICAM-1, the expression of iNOS mRNA and NF-κB protein in small intestine (jejunum and ileum) were significantly decreased [plasma LPS (kEU/L): 654.09±28.13 vs. 1 155.65±47.15, TNF-α (ng/L): 12.75±0.47 vs. 30.61±0.71, IL-1β (ng/L): 53.06±5.32 vs. 64.47±2.61; jejunum TNF-α(ng/L): 43.27±1.20 vs. 64.82±2.09, IL-1β (ng/L): 326.38±14.47 vs. 535.22±13.48, MPO (U/g): 0.14±0.01 vs. 0.32±0.02, iNOS mRNA (2-ΔΔCt): 2.39±0.13 vs. 10.80±0.22, NF-κB protein (gray value): 0.687±0.062 vs. 1.404±0.046; ileum TNF-α (ng/L): 62.75±3.92 vs. 104.24±2.82, IL-1β(ng/L): 408.06±1.70 vs. 521.97±1.16, MPO (U/g): 0.36±0.08 vs. 0.66±0.05, iNOS mRNA (2-ΔΔCt): 1.65±0.11 vs. 3.59±0.29, NF-κB protein (gray value):0.830±0.114 vs. 1.609±0.051, allP< 0.05].Conclusion KB can combine with LPS and inhibit LPS/Toll-like receptor 4 (TLR4) signaling pathway, thereby significantly inhibit the inflammatory response and protect the function of the small intestine in LPS-induced septic mice.

Objective To investigate the inhibitory effect of kukoamine B (KB) on lung inflammatory responses in mice with sepsis and its possible molecular mechanism.Methods Twenty-eight male mice were randomly divided into control group (n=8),lipopolysaccharide (LPS) group (n=10),and LPS + KB group (n=10).Sepsis model was reproduced by intra-peritoneal injection of 20 mg/kg LPS,while equivalent normal saline was given in control group,and 20 μg/kg KB was injected through caudal vein 4 hours after LPS challenge in LPS + KB group.After 8 hours of LPS challenge,the concentration of LPS in plasma and the activity of myeloperoxidase (MPO) in the lung tissue were determined.The contents of tumor necrosis factor-α (TNF-α) and interleukin-lβ (IL-1β) in plasma,alveolar lavage fluid and lung tissue homogenates were assessed by enzyme linked immunosorbent assay (ELISA).The activation of nuclear factor-κB (NF-κB) and the expression of inducible nitric oxide synthase (iNOS) in lung tissue were determined by Western Blot.The pathological changes in lung tissues were observed with hematoxylin-eosin (HE) staining.The expression of intercellular adhesion molecule-1 (ICAM-1) in lung tissue was determined by immunohistochemistry.Results Compared with control group,the concentration of LPS in plasma (kEU/L:1 155.650 ± 147.149 vs.31.390 ± 18.859),MPO activity (U/g:1.177 ±0.093 vs.0.775 ±0.166),NF-κB activity (gray value:1.557 ±0.105 vs.0.824 ±0.032) and the expression of iNOS (gray value:0.650 ±0.129 vs.0.392 ±0.097) were significantly increased in LPS group (all P＜0.05).After KB intervention,the concentration of LPS (624.461 ± 149.012),MPO activity (0.919 ±0.023),NF-κB activity (1.127 ±0.074) and the expression ofiNOS (0.425 ± 0.066) were significantly lowered (all P＜0.05).Compared with control group,the contents of TNF-α (ng/L:47.325 ± 13.864 vs.6.534 ± 0.544,13.382 ± 2.231 vs.3.748 ± 0.692,31.127 ± 7.399 vs.14.948 ± 4.673) and IL-1β (ng/L:74.329 ± 11.890 vs.29.921 ± 6.487,9.422 ± 2.674 vs.1.105 ± 0.364,528.509 ± 32.073 vs.109.945 ± 13.561) in plasma,alveolar lavage fluid and lung tissue homogenates were obviously enhanced in LPS group (all P＜0.05).With KB intervention,the contents of TNF-α (20.331 ± 7.789,7.145 ± 1.202,15.966 ± 2.946) and IL-1β (57.707 ±8.098,2.212 ± 0.878,426.154 ± 11.270) were markedly reduced (plasma TNF-α:F=16.052,P=0.002; IL-1β:F=20.649,P=0.000; lung tissue homogenates TNF-α:F=31.134,P=0.001; IL-1β:F=22.792,P=0.002;alveolar lavage fluid TNF-α:F=10.013,P=0.009; IL-1β:F=319.857,P=0.000).In addition,leukocyte infiltration to the lung tissue was attenuated,and the expression of ICAM-1 was reduced by KB in histological examination.Conclusion KB,as a neutralizer of LPS,can inhibit the release of inflammatory mediators,reduce the pulmonary inflammatory response and protect the function of lung in septic mice.

Objective To evaluate the clinical value of bedside index for severity in acute pancreatitis (BISAP) and APACHEⅡ score in predicting the severity and organ failure of acute pancreatitis (AP).Methods One hundred eighty-five patients of AP admitted to Department of Gastroenterology of First affiliated Hospital of Soochow University from January 2012 to December 2014 were studied retrospectively.According to BISAP score, patients who were ≥3 points were considered as high risk group, while ＜3 points were considered as low risk group.According to APACHEⅡ score, patients who were ≥ 8 points were considered as high risk group, while ＜ 8 points were considered as low risk group.According to the criteria of Pancreatic Diseases Group of Chinese Society of Gastroenterology of Chinese Medical Association, the patients were diagnosed as mild acute pancreatitis (MAP), moderately severe acute pancreatitis (MSAP), and severe acute pancreatitis (SAP).The BISAP, APACHEⅡ scores were calculated and compared between MAP group and MSAP + SAP group, respectively.The incidence of MSAP + SAP between high risk group and low risk group was also compared.The area of ROC curve (AUC) was used to evaluate the ability of BISAP and APACHEⅡ scoring system for predicting the severity of AP and the multiple organ dysfunction syndromes (MODS).Results Among 185 patients, MAP was identified in 101 patients, MSAP in 76 patients and SAP in 8 patients.Twenty-five MSAP patients developed organ dysfunction, and all the 8 SAP patients developed organ dysfunction.The BISAP scores of MSAP + SAP group and MAP group were (1.43 ± 0.89), (0.38 ± 0.61),andAPACHⅡ scores were (2.45± 1.36), (0.87± 0.62), the scores of MSAP+ SAP group were significantly higher than those in MAP group (P ＜0.01).In the 137 patients of low risk BISAP group, there were 47 MSAP + SAP patients (34.3％), while in the 48 patients of high risk BISAP group, there were 37 MSAP + SAP patients (77.0％);in the 153 patients of low risk APACHEⅡ group, there were 56 MSAP + SAP patients (36.6％), while in the 32 patients of high risk APACHEⅡ group, there were 28 MSAP + SAP patients (87.5％);the incidence of MSAP + SAP patients was significantly higher in high risk group than that in low risk group (P＜0.01).The AUC of BISAP, APACHEⅡ for MSAP+ MAP was 0.804 (95％ CI 0.738 ～ 0.870), 0.794 (95％ CI 0.725 ～ 0.863), and the AUC for organ dysfunction was 0.758 (95％ CI 0.686 ～0.830), 0.781 (95％ CI 0.710 ～0.852) , and the difference between BISAP and APACHE Ⅱ was not statistically significant (P ＞ 0.05).Conclusions The BISAP has the prediction ability for AP severity and prognosis similar to APACHEⅡ , and it consists of only 5 parameters and can be completed in the first 24 h of admission, therefore it is worth of clinical application.

Objective To investigate the microRNAs expression profile in human colorectal cancer with or without liver metastasis and try to screen miRNA associated with liver metastasis in colorectal cancer. Methods Twenty five surgical resected colorectal cancer specimens were collected and frozen in liquid nitrogen. Three without liver metastasis and three with liver metastasis were selected, from which total RNAs were isolated. The expressions of miRNAs in these two types of specimens were detected by illumine microRNA microarray, and the difference of miRNA expression was screened. The biochip results were verified with real-time RT-PCR in all colorectal caner specimens. Results The miRNA expression was significantly different in colorectal cancer with liver metastasis and without liver metastasis. Compared with colorectal cancer without liver metastasis, 28 miRNA expressions was different in colorectal cancer with liver metastasis, 4 up regulated and 24 down regulated. The quantity of miR-139-3p expression in colorectal cancer with liver metastasis was 1.75±0.40, up regulated compared with that incolorectal cancer without liver metastasis(0. 69 ±0.58,P＜0.05). The quantity of miR-19a expression in liver metastasis was 0. 39±0. 20, downregulated compare with no liver metastasis( 1.38 ± 0.98, P＜0. 05). The result of miRNA biochip was consistent with that of RT-PCR. Conclusion The difference of miRNA expression might relate to liver metastasis of colorectal cancer. The specific miRNAs expression profile might provide new target for diagnosis and treatment of colorectal cancer with liver metastasis.