Objective To investigate the potential role of MCP-1/CCL2 in experimental acute necrotizing pancreatitis (ANP) and complications. Methods 60 SD male rats were randomly divided into 3 groups: sham operation group ( n = 20 ), ANP group ( n = 20 ) and MCP-1 group ( n = 20 ). ANP model was induced by retrograde infusion of 3.5% sodium taurocholate, MCP-1 group received subcutaneous injection of MCP-1 antibody 0 h and 6 h after ANP induction. The serum levels of amylase, MCP-1, D-lactic acid,histological changes and the expression of MCP-1 mRNA of lung, small intestine and pancreas, the expression of MCP-1 protein in pancreas, MPO levels of small intestine MPO were determined. Results The serum levels of amylase, MCP-1, D-lactic acid in MCP-1 group at 12 h were (4666 ±412)U/L, (39.53 ±8.25)pg/ml and (6.3 ±2.2)mg/L, which were significantly lower than those in ANP group [ (9611 ±363)U/L, (63.42 ±9.32) pg/ml, (9.3 ± 2. 1 ) mg/L, P< 0.05 ) ]; the expression of MCP-1 mRNA in pancreas, small intestine and lung were 0.431 ± 0.009, 0. 211 ± 0.018 and 0.442 ± 0.017, which were significantly lower than those in ANP group [ (0.624 ±0. 010, 0. 523 ±0. 019 and 0. 569 ±0. 024, P <0.05) ]; the expression of MCP-1 protein in pancreas was 2.0 ± 0. 1, which was significantly lower than that in ANP group (4. 0 ± 0. 2, P <0.05). Lung and small intestine MPO were (11.1 ±3.0)U/g and ( 19.2 ±2.0)U/g, which were significantly lower than those in ANP group[(39.2±3.1)U/g and(13.1±2.1)U/g, P<0.05]. Conclusions Early blockade of MCP-1 not only attenuates the severity of ANP, but also decreases the degree of acute lung injury and intestine barrier dysfunction.

AIM: To investigate the effects of the dominant-negative I?B? plasmid on the expression of NF-?B and cyclooxygenase-2 (COX-2) after its being transfected into pancreatic carcinoma PC-3 cell line. METHODS: The expression of NF-?B and COX-2 in PC-3 cell line was confirmed by immunohistochemistry. The effects of dominant-negative I?B? plasmid transfection on the expression of NF-?B and COX-2 were studied by reverse transcription polymerase chain reaction (RT-PCR) and Western blotting. RESULTS: Both NF-?B and COX-2 were expressed in pancreatic carcinoma PC-3 cell line, and the expression of NF-?B and COX-2 were down-regulated in a certain time-independent manner after dominant-negative I?B? plasmid transfection. CONCLUSIONS: The NF-?B and COX-2 are expressed in pancreatic carcinoma PC-3 cell lines. The expression of NF-?B and COX-2 are inhibited by dominant-negative I?B? plasmid, while NF-?B is likely to play an important role in regulating the expression of COX-2.

Objective To investigate the expression of Iroquois homeobox protein 1 ( IRX1 ) gene and the hypermethylation status of its promoter in pancreatic cancer,and their relationship.Methods Real-time PCR was used to quantitatively detect IRX1 gene expression level of 12 sets of resected pancreatic cancer tissue and 6 sets of pancreatic cancer cell lines; gene sequences analysis was used to detect the structure of IRX1 promoter; DNA methylation inhibitor 5-Aza-2′-deoxycytidine (5-Aza-dC) was used in pancreatic cancer cell lines,and then the methylation of IRX 1 was measured by methylation-specific PCR (MSP) and unmethylation sequence-PCR (USP) methods.Results Expression of IRX1 mRNA in pancreatic cancer tissue was 0.31 ± 0.11,which were significantly lower than that in normal pancreatic tissue ( 1.05 ±0.32,P ＜0.01 ).IRX1 mRNA expression of AsPCl,BxPC3,Capan 2,PANCl,PaTu8988 and SW1990 were 0.36 ± 0.08,0.34 ±0.16,0.37 ±0.11,0.25 ±0.06,0.31 ±0.04,0.36 ±0.02,which were significantly lower than that in human kidney epithelial 293 cells ( 1.03 ± 0.28,P ＜ 0.05 or ＜ 0.01 ).Analysis of IRX1 gene sequence showed abundant CpG islands in promoter.Hypermethylation of IRX1 promoter site was found in all pancreatic cancer cell lines.However,its methylation status could be reversed by 5-Aza-dC,and the IRX1 expression was also restored.Conclusions IRX1 mRNA expression is down-regulated in pancreatic cancer,and it is related with promoter CpG islands hypermethylation.

Objective Metabonomics method based gas chromatography-mass spectrometry (GC/MS)were used to analyze the urine samples of patients with hyperlipidemic pancreatitis (HLP) to describe the characteristics of metabolism changes of HLP,identify potential biomarkers,and investigate the role of metabonomics study in the management of AP.Methods 24 patients of HLP and 40 age,sex matched volunteers were enrolled and their urine samples were collected.The urine samples underwent preparation,derivation and GC/MS analysis,Orthogonal-Projection to Latent Structures-Discriminant Analysis (OPLS-DA)were performed to detect the metabolic profile difference between the HLP and control group.Results HLP patients can be precisely distinguished from healthy controls.21 metabolites (credibility ＞ 700 ) were identified using the reference compounds available in the libraries of NIST and Wiley.It was identified that levels of nicotinic acid,aconitie acid,citric acid,hippurie acid,hydroxyphenylacetic acid,hydroxyphenylpropionicacid were decreased,while the levels of tryptophan,tyrosine,tyramine,16-hexadecanoic acid,18octadecanoie acid were increased.It was also suggested that there was change in tricarboxylic acid cycle and gut bacterial flora,as well as fat metabolism and metabolism of amino acid.Conclusions There are differences between healthy controls and HLP patients in the term of GC/MS metabolic profiling,and the biomarkers in the metabolites could be found through metabonomics analysis,and the mechanisms of the metabolic changes could be explored.It was noted that the research of metabolites in the urine samples may be a useful tool to help diagnose and understand the pathogenesis of HLP.Metabonomics analysis is a promising research method.

Objective To investigate the mechanism of hypedipidemia on pancreas of rats by comparative proteomic analysis.Methods Ten male SD rats were randomly divided into two groups,the experimental group Was fed with high lipid forage and the control group Was fed with normal food.Pancreatic samples from the two groups were harvested six weeks later.Differential protein analysis Was performed using differential in-gel electrophoresis(DIGE),and characterizing the protein biomarkers using tandem mass spectrometry.Western blot Was used to confirm the expression of significantly changed proteins.Results Compared to the normal pancreas tissue,a total of 3 protein spot-features were found to be significantly increased and 11 significantly decreased in pancreatic samples with hyperlipidemia.Significantly increased proteins in hyperlipidemia pancreatic samples were arginaseⅡ,ribonuclease inhibitor and glyeine amidinotransferase,which increased by 2.19,1.82 and 1.12 fold,respectively.Significantly decreased proteins in hyperlipidemia group were tyrosyl-tRNA synthetase,alpha-amylase,triacylglycerol lipase,DJ-1protein,Cu,Zn superoxide dismutase,which dicreased by 2.48,2.37,1.85,1.73 and 1.65 folds,respectively.Western blot analysis revealed increased arginase Ⅱ levels and decreased alpha-amylase in pancreatic samples with hyperlipidemia.Conclusions Pancreas wag possibly injured by hyperlipidemia via increase of arginase Ⅱ.Decreased amylase and lipase may be the protection mechanism of pancreas.

Objective:To investigate the expression and clinical significance of cell division cycle 42 (Cdc42) and WASP family verprolin-homologous protein l (WAVE1) in non-small cell lung cancer (NSCLC). Methods:The expression of Cdc42 and WAVE1 was detected in 106 paraffin-embedded NSCLC tissues and 46 adjacent normal lung tissues (control group) using immunohistochemis-try. Results:The expression levels of Cdc42 and WAVE1 was distinctly higher in NSCLC than in the control group. The expression of Cdc42 in NSCLC significantly correlated with tumor differentiation, TNM stage, and lymph node metastasis (P<0.05). The expression of WAVE1 in NSCLC was significantly correlated with TNM stage and lymph node metastasis (P<0.05 or P<0.01). The expression of Cdc42 was significantly correlated with WAVE1 in NSCLC (r=0.469, P<0.01). The 3-year survival rates were significantly lower in the group with high Cdc42 expression (44.16%) than in the low expression group (72.41%;P<0.01). Similarly, the 3-year survival rates were significantly lower among patients with high WAVE1 expression (39.44%) than in those with low expression (77.14%;P<0.01). Lymph node metastasis and the common high Cdc42 and WAVE1 expression were independent prognostic factors for NSCLC. Conclu-sion:The Cdc42 expression is correlated with WAVE1 expression. They may act together and have an important function in NSCLC. The expression of both Cdc42 and WAVE1 in NSCLC tissue may be used as markers for assessing the clinicopathologic features and prognosis.

Objective To compare the value of bedside index for severity in acute pancreatitis (BISAP),Ranson score and Balthazar computed tomography severity index (CTSI) in predicting the severity and prognosis of acute pancreatitis (AP).Methods From 2005 to 2011 in Shanghai,the clinical data of 1004 AP cases from seven hospitals was collected and retrospectively analyzed.The value of BISAP score,Ranson score and Balthazar CTSI in predicting the severity and prognosis of AP were assessed with receiver operator characteristic (ROC) curve.Results Among 1004 patients,the main cause of AP was biliary disease (580 cases),about 57.77％.The incidence of pancreatic necrosis,mortality and SAP increased along with BISAP score.The risk of pancreatic necrosis in patients with CTSI ≥ three was significantly higher than that of ＜ three.The risk of pancreatic necrosis and SAP in patients with BISAP score ≥ two was significantly higher than that of ＜ two (OR:4.93,95％CI 3.62-6.70; OR 2.62,95％CI 1.59-4.31,respectively).There was no significant difference in the accuracy of predicting the progression and mortality of AP among these three score systems.However the sensitivity of BISAP score (OR:61.54,95％CI 35.09-87.99) in predicting the progression and mortality of AP was better than that of Ranson (OR:46.15,95 ％ CI 19.05-73.25) and CTSI (OR:46.15,95％CI 19.05-73.25).Conclusions BISAP score is easy to perform and when combined with CTSI,it helps to make the diagnosis and classification of AP in time,predict the prognosis accurately.Compared with Ranson score,BISAP score has higher clinical value.

Objective To investigate analgesic effects of the selective blocking of descending facilitation targeting μ opioid receptor positive neurons in the rostral ventromedial medulla ( RVM) in a rat model of bone cancer pain. Methods Forty-eight adult female Wistar rats weighing 180-200 g were randomly divided into 6 groups: group Ⅰ control ( n = 3) ;group Ⅱ bone cancer pain induced by intra-tibia inoculation of Walker 256 mammary gland carcinoma cells ( n = 9) ;group Ⅲ-Ⅵ received a single intra-RVM micro-injection of PBS (group Ⅲ), dermorphin (group Ⅳ) , saporin (group Ⅴ) and dermorphin-saporin ( group Ⅵ) respectively at 28 days before intra-tibia inoculation ( n = 9 each) . Starting from 3 to 20 days after intra-tibia inoculation, mechanical allodynia was assessed and recorded. The animals were sacrificed on 7, 14 and 20 days after intra-tibia inoculation, after repetitive non-noxious tactile stimulation of the hindpaw. The total number of Fos-positive neurons in the spinal dorsal horn was measured as a marker indicative of central sensitization. Results The animals developed nociceptive hypersensitivity after intra-tibia cancer cell inoculation in group Ⅱ -Ⅵ . Nociceptive hypersensitivity was significantly decreased during 4-7 days after the onset of nociception in group Ⅵ (dermorphin-saporin). The number of Fos positive neurons in bilateral spinal dorsal horn was significantly increased by intra-tibia inoculation of cancer cells in group Ⅱ-Ⅵ as compared with control group and was significantly lower at day 14 and 20 after inoculation in group Ⅵ (dermorphin-saporin) than in group Ⅱ - Ⅴ.Conclusion Selective blocking of descending facilitation targeting μ opioid receptor positive neurons in RVM can effectively reduce nociceptive hypersensitivity induced by intra-tibia inoculation of Walker56 mammary gland carcinoma cells.

【Objective】 To study the application effect of gel adsorbent tank in the production of human prothrombin complex concentrate(PCC). 【Methods】 Six batches of PCC were produced from 1000 L cryoprecipitated plasma, using the same gel twice for adsorption within the tank.The number of gel repeated application was examined by retrospective confirmation, and the adsorption rate, specific activity and residue of finished virus inactivation reagent were determined before and after adsorption. 【Results】 All 6 batches of PPC, produced by the same gel, satisfied quality criteria. Both PPC solution and the gel presented good color. The average activities of coagulation Factors Ⅱ, Ⅶ, Ⅸ and Ⅹ of six batches of PCC were 118.2%, 157.0%, 140.5% and 176.8%, respectively. The The adsorption capacity of coagulation factor Ⅱ, Ⅸ and Ⅹ were both 100% in the first and second adsorption, while coagulation factor Ⅶ were 75% and 81%, respectively. The average specific activity of coagulation factor Ⅸ was 0.7 IU/mg. The average residues of polysorbate 80 and tributyl phosphate products were 0 μg/mL and 33 μg/mL, respectively. The same batch of gel can be repeatedly used up to 6 times during the PCC process. 【Conclusion】 The gel adsorption tank presents good application value in the production of PCC, which can realize process amplification and automatic control.

Rats ; Animals ; Urinary Bladder Neck Obstruction/pathology* ; Survivin/genetics* ; Urinary Bladder/pathology* ; Fibrosis