Abstract:Abstract:Objective:To analyze and compare the effect of early minimally invasive surgical treatment and conventional treatment for patients with severe acute pancreatitis.
Methods :According to the including criteria, eight randomized-controlled trials of this topic were enrolled into the analysis. The detail about the trial design, characters of the subjects and results of the studies were reviewed and analyzed by using Revman 4.2 software.
Results:Compared with conventional treatment, early minimally invasive surgical treatment was associated with a significantly lower incidence of mortality(RR=0.35,95%CI:0.20~0.63,P＜0.01), complications (RR=0.46, 95%CI:0.33~0.63,P＜0.01) and hospital stay duration (WMD=-22.47,95%CI:-35.41 ~ -9.53, P＜0.001), and with a higher cure rate (RR=1.24,95%CI:1.13～1.36,P＜0.01).
Conclusions:The findings suggest that for early severe acute pancreatitis, minimally invasive surgical treatment, should be selected as the first choice of treatment, while conventional treatment was used.

Abstract:Abstract:Objective:To study the selection of method and timing of operation in the treatment of acute biliary pancreatitis(ABP)
Methods :The clinical records of 236 cases of ABP from our hospital were reviewed and analyzed.
Results:All types of ABP underwent early or delayed operation, respectively.Mild pancreatitis patients were cured and discharged. For severe acute pancreatitis with bile duct obstruction, early surgery had no mortalily, but delayed operation mortality was 14.29%（1/7）. The mortality in patients with severe pancreatitis without bile duct obstruction who underwent early operation was 7.14%(1/14), and for those who had delayed operation was 14.29%(1/7). The complicalion score between the various groups was not statistically significant (P>0.05). The mortality rate for early operation of fulminant pancreatitis was 20.00%(1/5), and another case died after delayed operation.
Conclusions:In cases of ABP, early operation should be performed after aggressive non-surgical treatment for 24-48 h. Selection of operative method, according to the patient′s condition, can improve cure rate and decrease mortality rate.

Abstract:Abstract:Objective:To investigate the relationship between inducible nitric oxide synthase (iNOS ) mRNA expression of pancreatic tissues and intestinal permeability, and the effect of salviae miltiorrhizae on the change in gut permeability in rats with severe acute pancreatitis(SAP).
Methods :All rats were randomly divided into three groups: Sham operation（group C, n=15），SAP group（group SAP, n=15），and salviae miltiorrhizae therapeutic group （group T, n=15）. At 24h After operation, the iNOS mRNA expressions of pancreas were determined by in situ hybridization (ISH), and blood amylase(AMY), nitric oxide (NO) and lipopolysaccharide(LPS) were examined. The histopathologic changes of pancreas and ileum were observed. Intestinal permeability was measured by means of albumin clearance( AC)of 125I-labeled rat serum albumin.
Results:The iNOS mRNA expressions of pancreas in group SAP were markedly increased in comparison with group T (P＜0.01), while there was no iNOS mRNA expression of pancreas in group C. The blood AML, NO, LPS and ascites and intestinal permeability in group SAP were much higher than those of group C(P＜0.01),and also higher than those of group T (P＜0.01) except AC. The pathological changes of pancreas and ileum in group SAP were more severe than in group T(P＜0.01), however pathology of pancreas and ileum was normal in group C.
Conclusions:Excess NO produced by pancreatic iNOS mRNA overexpression may be a factor related to intestinal permeability in SAP. Salvia miltiorrhiza can down-regulate the iNOS mRNA expression of pancreas and improve the intestinal permeability in SAP.

Abstract:Abstract:Objective:To explore the role of apoptosis and the expression of X-linked inhibitor of apoptosis protein (XIAP) in polymorphonuclear neutrophils (PMNs) during acute pancreatitis (AP).
Methods :Blood from normal control (NC, n=15), mild acute pancreatitis (MAP, n=15) and severe acute pancreatitis (SAP, n=15) were collected. PMNs apoptosis was detected by flow cytometry. PMNs were isolated from each group and XIAP mRNA and protein levels were assessed by RT-PCR and Western Blotting.
Results:PMNs apoptosis in SAP group was (2.15±0.40)%, MAP group was (4.16±0.14)%，NC group was (4.31±0.12)%. PMNs apoptosis rate in SAP and MAP groups was decreased compared to NC group (P<0.05), in SAP group was markedly decreased compared to MAP group (P<0.01). The expression of XIAP in SAP and MAP groups was increased as compared to NC group (P<0.01), and XIAP expressed in SAP was markedly increased compared to MAP group (P<0.01).
Conclusions:XIAP may contribute to the severity of acute pancreatitis through suppression of apoptosis of PMNs.

Abstract:Abstract:Objective:To explore the possible mechanism of the protective effect of breviscapine on brain tissue in rats with acute hemorrhagic necrotizing pancreatitis, and to provide theoretical basis for the clinical treatment and prevention of pancreatic encephalopathy.
Methods :A total of 90 male SD rats were randomly divided into three groups with 30 rats for each group: normal control group (C group), acute hemorrhagic necrotizing pancreatitis group (AHNP group) and AHNP treated with breviscapine group(T group).The acute hemorrhagic necrotizing pancreatitis model in rats was induced by retrograde injection of 5％ sodium taurocholate into the pancreatobiliary duct. The brain samples were obtained in each group after the rats were killed at 6，12 and 24 hours respectively after operation. The activity of NF-κB and PKC-α in brain tissue was detected with immunohistochemistry and the expression of PKC-α mRNA in brain tissue was detected with RT-PCR. The level of serum TNF-α was measured by radioimmunoassay (RIA) and the changes of brain water content were measured.
Results:In T group, the activity of PKC-α and NF-κB, the expression of PKC-α mRNA in rat brain tissue, the serum TNF-α level and the changes of brain water contents were decreased significantly compared to the AHNP group(P<0.05).
Conclusions:Breviscapine can reduce inflammatory mediators by suppressing the activity of PKC-α, and then alleviate the acute pancreatitis-associated brain injury.

Abstract:Abstract:Objective:To investigate the effects of oxamate on concentrations of calcium in pancreatic cancer cells.
Methods :Pancreatic cancer cell panc-1 was cultured in vitro with oxamate for 48 h, then stained with Fluo-3/AM, and the light density of cells for different concentration of oxamate under confocal laser microscopy was observed.
Results:Oxamate influenced the concentration of calcium in panc-1,and this effect corresponded to the concentration of oxamate.
Conclusions:Oxamate can induce the increase of the concentration of calcium of panc-1,and then influence the cellular pathophysiologic process.

Abstract:Abstract:Objective:To explore the method of isolation and biological analysis of tumor stem cell of pancreatic adenocarcinoma cell line PANC-1.
Methods :PANC-1 cells were cultured. Subpopulation cells which have properties of tumor stem cells were isolated according to the cell surface markers CD44 and CD24 by flow cytometry from pancreatic adenocarcinoma cell line PANC-1. The obtained CD44-CD24-, CD44+CD24+ and unsorted PANC-1 cells were cultured in serum-free medium (SFM). The SFM was DMEM-F12 supplemented with 10 ng/mL fibroblast growth factor, 20 ng/mL epidermal growth factor, 5 kg/mL insulin, 2.75 mg/mL transferrin, 2.75 ng/mL selenium (insulin-transferrin-selenium solution), penicillin(1×105 U/L) and streptomycin (100 mg/L). Then the proliferative capability of these cells in vitro was estimated by MTT method. The tumor growth from different subpopulation cells inoculated into nude mice was studied, and expression of CD44 and CD24 of the CD44+ CD24+ cells -formed nodules and PANC-1 cells were detected by avidin-biotin-peroxidase complex (ABC) immunohistochemical staining.
Results:Expressed the cell surface marker CD44, in 5.1% to 17.5% of sorted PANC-1 cells, 21.8% to 70.1% expressed CD24, and only 0.9% to 3.5% of cells were CD44+CD24+. Compared with CD44-CD24- cells, CD44+CD24+ cells had a lower growth rate in vitro. For the former, index growth trend appeared at 5th day, while the latter appeared at 7th day. The sorted cell population with the highest tumorigenic potential were those cells expressing CD44 and CD24.After implantation of 1×105 CD44-CD24-cells in nude mice, no tumor growth was evident at 12 weeks. In contrast, nude mice implanted with 5×103 CD44+ CD24+ cells had large tumors evident at 4 weeks(2/8), and at least a 20～50-fold increase in tumorigenic potential(P<0.05 or P<0.01). There was no obvious histological difference between the cells of the CD44+ CD24+ cells-formed nodules and PANC-1 cells.
Conclusions:CD44 and CD24 may be used as the cell surface markers for isolation of pancreatic cancer stem cells from pancreatic adenocarcinoma cell line PANC-1. Subpopulation cells CD44+CD24+ have properties of tumor stem cells.

Abstract:Abstract:Objective:To study the expression of the apoptosis inhibitor Livin gene and its relationship with the expression of Bcl-2, p53 proteins in pancreatic carcinoma.
Methods :The expression of Livin mRNA was investigated by reverse transcriptase polymerase chain reaction (RT-PCR) in pancreatic carcinoma tissue of 52 cases and normal pancreatic tissue of 12 cases. The expression of Bcl-2 and p53 was detected by immunohistochemical assay.
Results:The expression rate of Livin gene in pancreatic carcinoma tissue was 71.2%, which was obviously higher than that in normal pancreatic tissue (P＜0.01). There was relationship between Livin gene expression and the differentiation and lymph node metastasis of pancreatic carcinoma (P＜0.05). The expression of Livin gene was correlated with Bcl-2 expression, but was not with p53 expression.
Conclusions:Livin gene may play an important role in the pathogenesis of pancreatic carcinoma. The up-regulated expression of Livin and Bcl-2 may play synergetic roles in the process of carcinogenesis of pancreatic carcinoma. There was no relationship between Livin expression and dysfunction of p53.

Abstract:Abstract:Objective:To study the relationship between chemoresistance and the expression of survivin，induced by gemcitabine, in pancreatic carcinoma cell line PANC-1.
Methods :The pancreatic carcinoma cell line PANC-1 was cultured by RPMI 1640 and gemcitabine with the concentration of 1 μg /ml and 10 μg /mL respectively, and survivin expression was examined by RT-PCR and Western blot. The relationship of the survivin level of the cell and its chemoresistance to gemcitabine was analyzed.
Results:Tweenty-four and 48 hours after culture with gemcitabine concentration of 1 μg /ml and 10 μg /mL respectively, survivin mRNA level was increased (1.34±0.12),(2.40±0.17),and(3.33±0.20),( 4.41±0.18) folds, and the protein expression was enhanced (1.20±0.07),(1.48±0.19),and(2.90±0.04),(4.50±0.20) folds respectively. There was a significant difference between the two groups (P＜0.05).
Conclusions:The intensity of pancreatic carcinoma cell lines chemoresistance could be enhanced by the action of gemcitabine on the up-regulated expression of survivin.

Abstract:Abstract:Objective:To investigate the potential mechanism of antigen-specific CD4+CD25+ regulatory T cells on the suppression of rejection for allogenetic islet transplantation in vivo.
Methods :Islets with 8×105 antigen-specific Treg were allogeneically transplanted under the kidney capsule of streptozotocin-induced diabetic BALB/cByJ mice. Mean survival　time(MST), the ratio of CD4+/CD8+ T cells, cytokine expression in both tolerant and rejected mice was detected.
Results:In vivo, the mean survival time of recipients with islets and antigen-specific CD4+CD25+ Treg group (C group) was (34.57±17.15) days, whereas transplanted islets without Treg treatment (B group) survived a mean time of (10.60±1.82)d(P<0.01). The ratio of CD4+/CD8+ T cells from antigen-specific CD4+CD25+ Treg transplantation group was significantly lower than that from islet transplantation group (P<0.01). The mRNA expressions of IL-10, TGF-β were upregulated in antigen-specific CD4+CD25+ Treg transplantation group. However, the mRNA expressions of IL-1β, IL-2, IFN-γ were similarly upregulated in islet B group.
Conclusions:Allogeneic antigen-specific CD4+CD25+ Treg cells can prolong islet graft survival by the mechanism of affecting Th2/Th1 cells responses.

Abstract:Abstract:Objective:To study the effect of glutamine and arginine in rats with hepatic fibrosis.
Methods :Thirty male Wistar rats with hepatic fibrosis induced by CCl4 for 6 weeks were randomly divided into control group(n=10)、glutamine treatment group(n=10) and arginine treatment group(n=10). At 8 weeks after treatment, the liver appearance、collagen protein level and the degree of hepatic fibrosis were observed.
Results:At the end of 8 weeks,there was no death of any group of the rats.The glutamine group showed the most severe hepatic fibrosis, the control group showed less, and the arginine group showed the least fibrosis.
Conclusions:The degree of hepatic fibrosis is aggravated by glutamine, while arginine obviously relieves rat hepatic fibrosis.

Abstract:Abstract:Objective:To investigate the effect of glucagon-like peptide-1(GLP-1) in a dose related manner on glucose metabolism after 65% hepatectomy in rats.
Methods :We determined the serum glucose levels of hepatomized rats at 0, 5, 10, 20, and 30 minutes after an intravenous glucose load (IVGTT, 0.5 g/kg glucose) on the first postoperative day, and the changes of blood glucose, serum insulin and glucagon concentrations of hepatomized rats that received the volum load with normal saline or 0.3 nmol/kg GLP-1, 0.45 nmol/kg GLP-1 respectively. Blood was drawn for determination of glucose (glucose oxidase), insulin, glucagon, and GLP-1 (radioimmunoassay).
Results:The peak glucose and 30-minute glucose levels and the area under the curve (AUC 0-30) were significantly higher in the hepatomized rats compared to the control rats, which had not undergone any operation and received a same intravenous glucose load (0.5 g/kg glucose with normal saline)(P<0.05), but were not significantly different compared to the rats that received the same volum load with 0.3 nmol/kg GLP-1 after liver resection (P>0.05). Nevertheless the peak glucose and 30-minute glucose levels and AUC 0-30 of the hepatomized rats that received with 0.45 nmol/kg GLP-1 were significantly lower compared to the rats that received the same volum load with normal saline or 0.3 nmol/kg GLP-1 respectively after liver resection. There was an increasing postoperative serum concentration of glucose, insulin, glucagon on the first day, then, the serum glucose concentration was significantly lowered after infusion of GLP-1 in rats undergoing hepatectomy (P<0.01), which might reach the glucose range in controls in a dose-dependent manner. Lowering of blood glucose was achieved by a significant rise of insulin secretion (P<0.05) and a suppression of glucagon release (P<0.05), with a potent stimulation of insulin secretion with infusion of 0.45nmol/kg GLP-1 on the first postoperative day.
Conclusions:In the early stage after hepatectomy in rats, the effect of GLP-1 in increasing insulin secretion and suppressing glucagon release were decteased, and increase of its dosage can augment these effects and further improve utilization of glucose.

Abstract:Abstract:Objective:To explore the effects of different preparation in adherent cells on the immunohistochemical display of membrane protein under light microscope.
Methods :Anchorage-dependant cells were cultured which included three groups: Gastric epithelial cell GES-1, gastric cancer cell SGC7901 and gastric cancer cell SGC7901/VCR. The cells were seeded on coverslips or cytospinned on the microscopic slides, and the MDR-1 detected by immunohistochemical method. Some of the specimens were re-stain with methyl green for afterstain. Finally, semiquantitative analysis was carried out.
Results:The MDR-1 immunoreactivity(IR) of specimens with re-stain was higher than that without re-stain in the same cell line and the same preparation, no matter whether the cells were seeded on coverslips or cytospinned on the microscopic slides. As to the same cell line, no matter whether afterstain or not, the MDR-1 IR in the cells cytospinned on the microscopic slides was higher than that in the cells seeded on coverslips. The MDR-1 expression level was different among three group cells, but their tendency of change was consistent in the same preparation and the same re-stain.
Conclusions:The adherent cells cytospinned on the microscopic slides have much better immunohistochemical display of membrane protein than those seeded on coverslips under light microscope.

Abstract:Abstract:Objective:To establish the standard method to extract total protein of pancreatic tissue by using different methods of pancreatic tissue protein extraction in rats.
Methods :The total protein extract kit and the author self-made lysate were used to extract the total protein of pancreatic tissue, then the protein level was measured and the protein was analysed by matrix -assisted laser desorption ionization-time of flight-mass spectra( MALDI-TOF-MS) respectively.
Results:The amount of total protein extracted with total protein extract kit and self-made lysate was （19.5±1.3 ）mg and （22.2±1.8 ）mg respectively. The MALDI-TOF-MS protein spectra peak was (404.4±13.7) and( 437.2±18.1) respectively.
Conclusions:The self-made lysate extraction method has higher protein extraction rate and better protein dissolubility as compared with total protein extract kit extraction method.

Abstract:Abstract:Objective:To study the diagnosis, treatment and prognostic factors of solid and pseudopapillary tumor of the pancreas (SPTP) in domestic.
Methods :Eight cases with SPTP in our hospital and a review of 397 others from the literature of China were analyzed. Clinical, radiographic and pathologic features were collected, and influence facters of prognosis were evaluated
Results:The patients (91.36% women) were of mean age 26.76 years; 40.99% of the patients were without subjective symptoms. CT was more precise than ultrasonography in determining tumor nature and location. The tumors were evenly distributed in the head/neck and body/tail of the pancreas. Surgery was the main therapeutic approach; 3.71% patients developed postoperative pancreatic fistula usually after local resection. Pathology was the definitive diagnostic method. Most tumors were positive for vimentin, α-antitrypsin and α-antichymotrysin, while negative for insulin, glucagon and CEA. Disease-free survival of mean 23.5 month was achieved in 95.40% cases after initial or second surgery. Tumor advanced in 5.02% of the patients. In our center, 62.50% of cases had been infected by HBV.
Conclusions:SPTP mainly occurred in young females and is a potentially low-malignancy tumor. Preoperative diagnosis is made through CT exam and postoperative diagnosis is confirmed by immunochemistry and pathology. Surgery is the predominant and effective method of treatment. Probable prognostic factors include gender, surgical modality and integrity of tumor capsule.

Abstract:Abstract:Objective:To summarize our experience in diagnosis and treatment of splenic tumors.
Methods :A retrospective study of the clinicopathologic data of 92 cases of splenic tumor was performed.
Results:There were 47 benign and 45 malignant tumors. The preoperative confirmed diagnostic rate by B-US was 85.87%(79/92), by CT was 91.30%(84/92). Splenectomy was performed in 81 cases, tumor resection or partial splenectomy in 7 cases, needle aspiration of splenic cyst in 3 cases and biopsy in one case. The 1-, 3-, 5-, 10-year survival rate of malignant tumor was 68.29%,31.70%,14.63%,0.24%,respectirely.
Conclusions:Imaging studies are the main diagnostic methods of splenic tumors. Splenectomy is the treatment of choice for primary splenic malignant tumor. Splenic benign tumor does not require any other therapy after operation. Splenic malignant tumor requires adjuvant treatment.

Abstract:Abstract:Objective:To investigate the factors of influence in post-traumatic stress bleeding.
Methods :The clinical data of 1986 trauma patients and postoperative patients in recent 5 years were studied retrospectively, and cases with or without stress ulcer bleeding were comparatively analyzed.
Results:The incidence of stress ulcer bleeding in cases with serious injury was significantly higher than in those with mild injury (P＜0.01).The incidence of stress ulcer bleeding in patients with diabetes, heart and lung disease, and patients ≥60 years old was significantly higher than those without diabetes, heart and lung disease, and patients ≤60 years old (P＜0.05）.
Conclusions:The post-traumatic stress ulcer bleeding is determined by combined factors including injury severity(ISS), patients′ age, underlying diseases and treatment methods.