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 investigate the effect of capsaicin on hepatic stellate cells (HSCs) and liver fibrogenesis.Methods HSCs were cultured.The reactive oxygen in HSCs under capsaicin at different concentrations was tested by DCFH-DA kit.The proliferation of HSCs was detected by CCK-8 test kit.Smoothmuscle α-actin (α-SMA) expression of HSCs was evaluated by Western blot.The fibrosisrelated genes were tested by RT-PCR.The apoptosis of HSCs was measured by flow cytometer.Bcl-2,bax and cyt-c was detected by Western blot.A murine model of liver fibrogenes was established.Capsaicin of different concentration was injected intraperitoneally.Liver pathology was observed using HE staining.Hydroxyproline content of liver and levels of collagen Ⅲ and hyaluronic acid in serum were tested.Results In dose dependent manner capsaicin inhibited the generation of the reactive oxygen species.Proliferation and activation of HSCs was inhibited by capsaicin (respectively F =13.267,57.392,all P ＜ 0.05) and the apoptosis of HSCs was promoted by capsaicin (F =235.571,P ＜ 0.05).Bax,cyt-c and caspase-3 was increased obviously (respectively F =29.334,38.274,138.329,all P ＜ 0.05).Capsaicin changed the expression of fibrosis-related genes (TGF-β1,TIMP-1) in HSCs (respectively F =376.534,253.751,all P ＜0.05).Capsaicin downregulated the level of hydroxyproline,collagen Ⅲ and hyaluronic acid in the rat model (respectively F =153.397,27.149,38.392,all P ＜ 0.05).Conclusions Capsaicin inhibits the proliferation and activation of hepatic stellate cells.Capsaicin promotes the apoptosis of hepatic stellate cells,and inhibits liver fibrogenesis.

Objective To investigate the effect of adipose derived stem cells (ADSCs) on hepatic stellate cells (HSCs) in vitro and on liver fibrosis in vivo.Methods ADSCs and HSCs were isolated from adipose tissue and liver respectively in SD rats.The coculture system was set up by transwell insert.The 5th passage HSCs were cultured on the 6-well plastic plate,and ADSCs or BRLs seeded on the transwell insert.The proliferation of HSCs was tested by CCK-8 test kit.Smoothmuscle α-actin (α-SMA) expression of HSCs were tested by Western blot.Rat models of liver fibrosis was established.Rats in ADSCs treatment group were infused with ADSCs and those in control group were infused with Buffalo rat liver cells (BRLs).Liver sections were studied by immunocytochemistry.Liver hydroxyproline (Hyp) content,serum laminin (LN)and hyaluronic acid (HA) were tested,the cytokines in the culture medium were assayed.Results HSCs and ADSCs were isolated successfully.After coculture for 72 h,compared with the control group,the proliferation and activation of HSCs was inhibited by ADSCs( absorbance of each group were 2.172 ±0.107,1.424 ± 0.013,1.209 ± 0.117,F =90.605,P ＜ 0.05 ; Gray-scale values of each group were 1.4 ± 0.2,152 ± 14,258 ± 18,F =283.348,P ＜ 0.05 ),ADSCs infusion inhibits liver fibrosis in model rats ( F =77.234,65.164,58.309,all P ＜ 0.05 ).More hepatocyte growth factor(HGF) and less transforming growth factor-β1 (TGF-β1) (F=1.767,P＜0.05)and nerve growth factor (NGF) (F=2.301,P＜0.05) were secreted by ADSCs than by BRLs.Conclusions ADSCs inhibit the proliferation and activation of hepatic stellate cells.Treatment with ADSCs decreases collagen deposition in the liver and inhibits liver fibrosis.

Objective To establish human pancreatic cancer xenografts in nude mice, and to investigate the antitumor efficacy of human endostatin expressed by replication-competent adenovirus AdTPHre-hE in vivo. Methods Pancreatic cancer cells AsPC-1 were injected subcutaneously in BALB/c nude mice to establish the xenografts. Tumor growth was observed and measured after AdTPHre-hE treatment. Expression of endostatin was detected by ELISA assay. The tumors were harvested for pathologic examination and immunohistochemical staining. Results Tumors grew more slowly in the AdTPHre-hE group and their sizes were markedly smaller than those of the Ad-hE group (P＜0.01)and control group(P＜0. 01). Endostatin levels were detected in the sera of nude mice in all treated groups, and endostatin expression in AdTPHre-hE group increased with time. The endostatin level in the AdTPHre-hE treated group was much higher(P＜0. 01)and increased faster than that in the Ad-hE treated group. Immunohistochemical staining for Hexon of adenovirus capsid showed more positive tumor cells in the tumor tissues treated with AdTPHre-hE. Immunohistochemical staining for FⅧ revealed a decreased microvessel density in the tumor tissues treated with AdTPHre-hE. Conclusion The replication-competent adenovirus efficiently expressed high-level endostatin and significantly inhibited tumor growth in vivo.

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 low bilirubin level on hepatic stellate cells (HSCs) in vitro. Methods HSCs were isolated and cultured from the liver of SD rats. The effect of bilirubin in different concentration on reactive oxygen in HSCs was determined by DCFH-DA kit. The proliferation of HSCs was tested by CCK-8 test kit.Smoothmuscle α-actin (α-SMA) expression of HSCs was tested by Western blot.The apoptosis of HSCs was tested by flow cytometry.The fibrosis-related genes were tested by PCR. Results HSCs were isolated and cultured successfully.Bilirubin in low concentration (0,1,10,20 mg/L) inhibited the generation of the reactive oxygen.Proliferation and α-SMA expression of HSCs was inhibited by bilirubin (0.624 ± 0.092,0.536 ± 0.127,0.407 ± 0.033,0.399 ± 0.022,F =13.454,P ＜0.05 ; 339 ± 2,336 ± 10,246 ± 7,242 ± 5,3.7 ± 0.3,F =191.107,P ＜ 0.05 ) and the apoptosis of HSCs was promoted by bilirubin(2.69 ±0.07％,2.95 ±0.10％,4.41 ±0.22％,4.91 ±0.86％,F =34.731,P ＜0.05 ).Bilirubin in low concentration changed the expression of fibrosis-related genes in HSCs.The ratio of TIMP-1mRNA/MMP-2mRNA decreased (54 ± 2,65 ± 2,47 ± 2,44 ± 2,F =73.400,P ＜ 0.05).Conclusions Bilirubin in low concentration inhibits the proliferation and activation of hepatic stellate cells,orobablv bv a mechanism in which bilirubin promoted antioxidative function.

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.