Objective To propose a rational clinical classification of gallstone pancreatitis for better guide and select clinical treatment scheme. Methods On the basis of severity of pancreatitis and the presence or absence of biliary obstruction, 273 cases of gallstone pancreatitis were classified into four types: non obstructive mild type (type Ⅰ) , obstructive mild type (type Ⅱ) , obstructive severe type (type Ⅲ) , non obstructive severe type (type Ⅳ). Moreover, according to the presence or absence of common bile duct stone, every type was further classified into two subtypes: subtype a and subtype b. Then, the results of clinical classification, treatment methods and prognosis were analyzed. Results Ⅰa subtype: 34 cases, Ⅰ b subtype: 112 cases; Ⅱ a subtype; 59 cases, Ⅲ subtype; 11 cases; Ⅲa subtype; 6 cases, Ⅲ b subtype: 4 cases; Ⅳa subtype: 3 cases, Ⅳb subtype: 44 cases. The overall mortality was 3.3% (9/273) , the mortality in Ⅰ type, Ⅱ type, Ⅲ type or Ⅳ type was 0, 0, 10% (1/10), 17.0% (8/47), respectively. The difference was statistically significant (P<0.05). The mortality of Ⅳ type in early operation group, traditional non-operative group, and regional intra-arterial infusion group was 30. 8% (4/13) , 25% (3/12) , 4. 5% (1/22) , respec tively. The mortality of regional intra-arterial infusion group was significantly lower than those in other two groups (P<0.05). Conclusions This 4 types and 2 subtypes classification method of gallstone pancreatitis was rational. The treatment efficacy may be improved according to the clinical classification. However, attention shall be paid to the transformation of these clinical types.

Objective To investigate the effect of liver function with portal vein occlusion (PVO) in various phases and the following restoration portal vein flow on intestinal mucosal cells. Methods Twenty-four healthy adults white Japanese rabbits were randomized into 1 control group and 2 experimental groups (according to portal vein clamping for 30 min and 45 min). Each experimental group's blood samples were collected from caval vein 1 h before operation, by the end of portal vein oc-cluded, 30 min, 60 rain after relief of portal vein blocking, then with restoration of portal vein flow for 2 h and rabbit guts were continuously cut to sections for HE, TUNEL staining and Bcl-2, Bax protein immunohistochemical staining to observe the injury of intestinal mucosa cell apoptosis and the relation-ship between the expressions of Bcl-2 and Bax. The levels of blood glutamate-pyruvate transaminase (ALT), glutamic oxalacetic transaminase (AST) were measured and compared. Results The levels of ALT, AST in the control group did not significantly change. Compared with control group, group B did not change significantly with PVO 30 rnin in liver enzyme and they were significantly increased after portal vein occlusion relief. The levels of ALT and AST were increased obviously at 45 min with PVO, then raised again. Down-regulation of Bcl-2 expression, up-regulation of Bax expression and the increased index of apoptotic cell were found in each experimental group. Conclusion It may be more safe with PVO for 30 min. But the following restoration portal vein flow will bring about ischemia-reperfusion injury that is mainly apoptosis in the small intestine. The index of apoptosis will be raised with time prolongation of PVO.

Objective To investigate the effect of hepatic stellate cells (HSC) on proliferation and invasion of hepatocellular carcinoma cells and the possible mechanism involved.Methods The HSC was isolated by optiprep method.Methyl thiazolyl tetrazolium(MTT) assay was used to detect the proliferation of hepatocellular carcinoma cells.The effect of invasion was measured with transwell assay.Matrix metalloproteinase-2 (MMP-2) and nuclear factor-κB (NF-κB) were detected by Western blotting.Results HSC was isolated and cultured successfully.HSC promoted the proliferation and invasion of hepatocellular carcinoma cells (proliferation:0.571 ±0.024 vs 0.803 ±0.048,1.271 ±0.044,1.973 ±0.036; invasion:25.2 ± 1.9 vs 35.8 ±3.3,44.4 ±2.7,53.9 ±3.6) (P ＜0.05).MMP-2 (1.32 ±0.22 vs 2.46 ±0.39) and NF-κB(0.85 ±0.09 vs 1.44 ±0.21) were increased obviously in hepatocellular carcinoma cells stimulated by HSC.Conclusions HSC can promote the proliferation and invasion of hepatocellular carcinoma cells.The mechanism might be related to up-regulation of the expressions of MMP-2 and NF-κB.

Objective To evaluate a new method for the isolation of rat pancreatic stellate cells (PSCs) and to investigate the influence of adipose derived stem cells (ADSCs) on PSCs in vitro.Methods Normal rat PSCs was isolated by collagenase and Optiprep density gradient centrifugation.The coculture system of ADSCs and PSCs was set up by transwell insert.The proliferation of PSCs was tested by CCK-8 test kit.Smoothmuscle α-actin (α-SMA) expression of PSCs were tested by Western blot.The apoptosis of HSCs was tested by flow cytometer.The cytokines in the culture solution were tested by ELISA kit.Results The quantity of PSCs was above 5 × 106 cells per rat.The purity and the viability of the cells were about 90-97 percent.After coculture for 72 h,the proliferation and activation of PSCs was inhibited by ADSCs (F =223.27,P ＜ 0.05 ; F =52.97,P ＜ 0.05) and the apoptosis of PSCs was promoted by ADSCs (F =43.62,P ＜ 0.05).more NGF and less TGF-β1 was secreted by ADSCs than by PSCs (NGF:14.68 ± 0.94 vs.8.31 ±0.86,t =4.67,P ＜0.05;TGF-β1:10.65 ±0.46 vs.70.47 ±0.99,t =21.72,P＜0.01).Conclusions ADSCs inhibit the proliferation and activation of PSCs by ADSCs secreting cytokines.

Objective To compare the difference between bone marrow stomal cell (BMSC) and adipose-derived stem cell (ADSC) of liver fibrosis in rats.Methods BMSC and ADSC of Sprague-Dawley (SD) rats were isolated and purified.The stem cell markers were detected with flow cytometry.The coculture system was set up with 0.4 μm Transwell insert semipermeable membrane.ADSC or BMSC were co-cultured with hepatic stellate cells (HSC).Normal hepatocyte cell line of rat (BRL) was co-cultured with HSC as negative control group and HSC cultured alone was blank control group.After cultured for 72 hours,the proliferation of HSC was determined by cell counting kit-8 (CCK-8) method.The expression of α-smooth muscle actin (α-SMA) of HSC was detected by Western blotting.The apoptosis of HSC was examined by flow cytometry.After BMSC,ADSC and BRL cultured alone for 72 hours,expression level vascular endothelial growth factor (VEGF),interleukin-10 (IL-10),nerve growth factor (NGF) and transforming growth factor-β1 (TGF-β1) in the culture medium were detected by enzymelinked immunosorbent assay (ELISA) method.The rats model of liver fibrosis were established.The rats were divided into BMSC treatment group,ADSC treatment group,BRL group and culture medium group,six rats in each group,which were injected with 1.5 mL BMSC,ADSC and BRL cells suspension (5 × 106) through portal vein,respectively,and same volume of culture medium was injected to the rate of culture medium group,once every two weeks for four weeks.The pathological changes of liver tissue sections were observed and liver fibrosis markers were tested.T test was performed for comparison between two samples and analysis of variance was used for comparison among multiple groups.Results BMSC and ADSC were successfully isolated and cultured.The phenotype of BMSC and ADSC was similar.Compared with blank control group and negative control group,both ADSC and BMSC could inhibit the proliferation of HSC and promote apoptosis (proliferation,2.43±0.27,2.39±0.33,1.92±0.38 and 2.18±0.31,FBMSC =25.61,FADSC =38.63,both P＜0.05 ;apoptosis rate,(5.59 ± 0.40)％,(6.82±0.57)％,(8.31± 1.03) ％ and (9.36 ± 0.54) ％,FBMSC =73.69,FADSC =97.41,both P＜ 0.05).The effects of ADSC were more significant than those of BMSC (t=5.76 and 5.18,both P＜0.05).There was difference in the cytokine levels secreted by ADSC and BMSC (NGF,(7.46 ± 0.54) pg/mL vs (3.95 ± 0.71) pg/mL,t =10.92,P＜0.05; TGF-β1,(8.79 ±0.93) pg/mL vs (6.36±0.85) pg/mL,t=7.58,P＜0.05).The cell transplantation experiment indicated that both BMSC and ADSC had significant inhibitory effect on liver fibrosis.The activity index of inflammation and degree of fibrosis in BMSC treatment group and ADSCs treatment group were 9.87±2.07,4.17 ± 0.94 and 10.13 ± 1.81,3.98 ± 0.82,which were significantly lower than those in blank control group (13.78±2.53 and 5.09±1.15)and negative control group (13.34± 1.89 and 4.95± 1.22,FBMSC=51.26 and 32.29,P＜0.05; FADSC =46.73 and 40.94,P＜0.05).The level of hyaluronic acid ((191.5±33.2) μg/L and (178.8±28.2) μg/L),type Ⅲ collagen ((19.9±5.1) μg/L and (21.7± 3.3) μg/L) and hydroxyproline ((312.6±38.8) μg/g and (325.8±28.2) μg/g) of BMSC treatment group and ADSC treatment group were significantly lower than those of negative control group and blank control group (hyaluronic acid,(282.3 ± 18.7) μg/L and (287.5 ± 26.7) μg/L),F =73.51 ; type Ⅲ collagen,(35.3± 3.3) μg/L and (32.5±4.3) μg/L,F=76.19; hydroxyproline,(458.4 ± 38.1) μg/g and (473.9 ± 63.7) μg/g,F=60.37,all P＜0.05).However,there was no difference between BMSC treatment group and ADSC treatment (all P＜0.05).Conclusions ADSC and BMSC had similar stem cell characteristics.There was difference in inhibiting the activation of HSC between ADSC and BMSC.But there was no significant difference in inhibiting liver fibrosis of rats in vivo.

Objective To study the clinical value of peritoneal cavity non-drainage after the operation of acute perforating appendicitis.Methods 196 patients with perforating appendicitis were randomly divided into drainage group and non-drainage group.The incidence rates of wound infection and ankylenteron and hospital durations in the two groups were observed and compared with each other.Results The incidence rate of wound infection and ankylenteron were 19.0%,10.7% in the drainage group and 8.0%,4.5% in the non-drainage group respectively(P0.05).The mean postoperative hospital stay of the drainage group was(9.3?2.7)days,which was significantly longer that of the non-drainage group(5.1?1.9)days,P

Objectives In this retrospective study we investigated the causes of misdiagnosis and incorrect treatment of hepatic cholangiocarcinoma and ways helpful in the improvement of correct diagnosis.Methods There were altogether pathollgy proved 40 cases of hepatic cholangiocarcinoma. The preoperative diagnostic procedures and surgical measurcs adopted were reviewed. Results The primary misdiagnosis rate was 68%. Patients were misdiagnosed as cholelithiasis complicated by intrahepatic inflammatory mass, hepatic abscess, hepatic hydrocyst, and hepatic adenoma;The surgical procedure performed were: choledocholithotomy, hepatophyma incision drainage or liver puncture drainage on B mode ultrasound localization, and hepatic cyst fenestration. Conclusions The hepatic cholangiocarcinoma can mimic many other benign diseases leading to misdiagnosis and improper surgery. Hence clinical features,history and laboratory evidence characteristic of the cancer must be sought preoperatively and intraoperative biopsy should be taken before definite surgical procedure. [

Objective To study the protective effects of resveratrol against hepatic stellate cells (HSCs) and liver fibrogensis.Methods HSCs were isolated from liver of SD rats.The reactive oxygen output in HSCs under resveratrol in different concentrations was tested by DCFH-DA kit.The proliferation of HSCs was tested by CCK-8 test kit.Smoothmuscle α-actin (α-SMA) expression of HSCs was evaluated by Western blotting.The activity-related genes were measured by PCR.The models of liver fibrogenes were established.Resveratrol in different concentrations was administrated intraperitoneally.Liver was studied by pathology and SMA staining.Hydroxyproline content of liver and levels of collagen Ⅲ and hyaluronic acid in serum were tested.Results HSCs were isolated from liver and cultured successfully.Resveratrol inhibited the generation of the reactive oxygen.Proliferation and activation of HSCs was inhibited by resveratrol (0.536 ±0.052,0.411 ±0.047,0.327 ±0.063,0.312 ±0.032,F =12.776,P ＜0.05) (103 ±7,90 ±7,63 ± 4,53 ± 3,F =62.179,P ＜ 0.05).Resveratrol inhibited the expression of genes (myogenic determination gene MyoD,collagen 11 and collagen Ⅰ) in HSCs(122 ± 5,96 ± 3,68 ± 3,60 ± 3,F =180.600,P＜0.05) (100±8,82 ±3,53 ±3,51 ±2,F=77.451,P ＜0.05) (170 ±3,147 ±4,92 ±3,90 ±2,F =462.878,P ＜ 0.05).Resveratrol downregulated the level of hydroxyproline,collagen Ⅲ and hyaluronic acid (358.3 ± 20.2,320.5 ± 15.3,290.3 ± 24.5,F =23.929,P ＜ 0.05) (32.8 ± 3.1,28.9 ±1.3,25.3±1.8,F=20.050,P＜0.05)(276.3 ±17.8,225.3 ±28.3,195.4 ±11.2,F=18.585,P＜0.05).Conclusions Resveratrol can inhibit the proliferation and activation of HSCs and downregulate the fibrogensis level of the liver of rats.

Objective To investigate the influence of fat-specific protein 27 (Fsp27) gene on the regulation of liver fibrogenesis in vivo.Methods Hepatic stellate cells (HSCs) were isolated from rat liver.Fsp27 gene was detected in primary HSCs and activated HSCs by real-time quantitative PCR (RTqPCR).Lentiviral vector carrying Fsp27 gene was constructed.The model of liver fibrosis was established by infusing carbon tetrachloride (CC14).The rats with liver fibrogenesis were infected by the virus.Liver sections were made to observe the structure and form of liver histocytes.The content of fibrous protein in liver and serum was detected by enzyme-linked immunosorbent assay (ELISA) and radioimmunoassay.Resukts HSCs were isolated and cultured successfully.The difference of Fsp27 gene between primary HSCs and activated HSCs was significant(P ＜ 0.01).The model of liver fibrosis was achieved.After infecting the model rats,we found the fibrosis level in treatment group was lower compared with control group.Conclusions Fsp27 treatment can decrease collagen deposition in the liver and inhibit the formation of fibrosis.

Objective To explore the effects of adipose tissue-derived stem cells (ADSCs) on the proliferation and invasion of pancreatic cancer cell lines in vitro.Methods ADSCs were isolated and co-cultured with pancreatic cancer cells.The proliferation and invasion of pancreatic cancer cells was tested by CCK-8 test kit.Stromal cell derived factor (SDF-1),vascular endothelial growth factor(VEGF),matrix metalloproteinase(MMP-9)and transforming growth factor-β (TGF-β1)in the culture medium were assayed by ELISA.The expression of cytokines in pancreatic cancer cells and ADSCs were detected by qRT-PCR.CCK-8 test kit was used to measure the AMD3100 regulation for the co-culture of ADSCs and pancreatic cancer cells.Results ADSCs can promote the proliferation and invasion of pancreatic cancer cells (all P ＜0.05); more SDF-1,VEGF,MMP-9 and less TGF-β1 was secreted by ADSCs than by pancreatic cancer cells(SDF-1:1100±100 vs.0 vs.0,F=389.134,P＜0.01;VEGF:140±4 vs.99±5 vs.93 ±4,F=174.102,P ＜0.05;MMP-9:61.8 ±4.2 vs.43.5 ±2.8 vs.54.5 ±3.0,F=76.279,P＜0.05;TGF-β1:20.6 ±3.0 vs.35.6 ±2.6 vs.41.3 ±5.5,F =79.338,P ＜0.05).ADSCs promote the expression of VEGF and MMP-9 in pancreatic cancer cells(VEGF:63.7 ±5.9 vs.50.6 ±4.1,t =7.536,P＜0.05; MMP-9:mRNA(55.8±3.6 vs.42.7 ±3.1,t =8.279,P＜0.05).AMD3100 significantly downregulates these growth-promoting effects of ADSCs on pancreatic cancer cells(SW1990:1.539 ±0.140 vs.1.361±0.066,t=2.835,P＜0.05;PANC-1:1.376±0.100 vs.1.281±0.031,t=2.860,P＜0.05).Conclusions ADSCs promote the proliferation and invasion of pancreatic cancer cells.