Objective To observe the effect of high mobility group box-1 protein (HMGB1) on the expression of intestinal epithelial tight junction protein occludin in murine severe acute pancreatitis (SAP).Methods Rat SAP model was estabilished by retrograde injection of 5 ％ sodium taurocholate into choledochopancreatic duct.Healthy wistar rats were divided randomly (random number) into three groups:control group,SAP group,pyrrolidine dithiocarbamate (PDTC) therapy group.Levels of plasm amylase,lipopolysaccharide (LPS) and D-lactate were determined.The changes of morphological damage of pancreasand intestinal tissues were observed by microscopy.The distribution and expression of occludin protein were observed by SP immunohistochemistry. The mRNA expression of HMGB1 in the intestinal mucosa was detected by reverse-transcription polymerase chain reaction (RT-PCR).The expressions of HMGB1 and occludin were determined by western blotting. One-way analysis of variance was performed with SPSS Windows 13.0 statistical analysis software,and a difference was accepted as significant if P ＜ 0.05.Results In comparison with the other two groups,levels of plasma LPS and D-lactate in SAP group increased markedly at 24 h after operation,which indicated that the penetrability of intestinal mucosal barrier increased (P ＜ 0.05 ). The expression of HMGB1 in the intestinal mucosa of SAP group increased significantly compared with control group (P ＜ 0.05).Whereas,the expression of occludin was significantly lower than control group (P ＜0.05).Compared with SAP group,the expression of HMGB1 was lower and the expression of occludin was higher in PDTC group ( P ＜ 0.05).Conclusions The over-expression of HMGB1 could down regulate the expression of occludin in intestinal tissues of SAP rats,and thus mediate an increase in penetrability of intestinal mucosal barrier.As PDTC inhibited the expression of HMGB1,the expression of occludin protein was up-regulated and the function of intestinal mucosal barrier was improved.

Objective To explore the effects of ethyl pyruvate (EP) on hepatic high mobility group box-1 protein (HMGB1) expression in experimental routine with acute necrotizing pancreatitis (ANT). Method ANP model was induced by retrograde injection of 5 % sodium taurocholate into pancreatic duct. Twenty-four male wistar rats were divided randomly into 3 groups(8 rats in each group): group A (ANT group); group B (ANP rats re-ceived ethyl pyruvate therapy) and group C (control group with sham operation). The concentration of plasma amylase (AMY), A.sr and ALT, and the activity of myeloperoxidase (MPO) in the liver were determined. The ex-pression of HMGB1 mRNA in liver was detected by using reverse transcription polymerase chain reaction (RT-PCR). The changes of morphological damage were observed under microscopy. The expression of HMGB1 in the liver was observed by using SP immunohistochemistry. ANOVA was performed with SPSS 10.0 statistical analysis software and the difference was accepted as significant if the P<0.05, as verified by using Duncan's and Tukey' s post hoc test. Results Compared with gxoup A,levels of plasma AMY,AST and ALT in group B were markedly lower (P<0.05). Compared with group C, MPO in group A was higher significantly (P<0.01).with group A, the pathological changes of pancreas and liver in group B were milder. Compared with group C,the hepatic HMGB1 mRNA expression was markedly higher in group A [(0.28±0.04) vs. (0.73±0.06), P<0.01]. By contrast,the HMGB1 mRNA expression was markedly lower in group B compared with group A [(0.46±0.05) vs. (0.73±0.06), P<0.05]. The HMGB1 protein expression in hepatocytes and Kupffer's cells of rats with ANP was significantly up-regulated compared with control group, but it was reduced significantly in EP treatment group. Conclusions HMGB1 as a late mediator in liver might be involved in the pathogenesis of acute hepatic injury with ANP. EP could down-regulate the hepatic HMGB1 expression together with improvement of liver function in rats with ANP.

OBJECTIVETo investigate the therapeutic efficacy of compound immunotherapy of tumor-derived heat shock protein 70 (HSP70) and interleukin-2 (IL-2) on tumor-bearing mice, and to provide reference for translating this strategy to human cancer.

METHODSCell culture, techniques for protein extraction and purification, SDS-PAGE, Western blot and capillary electrophoresis for HSP70 detection and purity analysis, and animal experiments were used. Mice were treated with HSP70 5 or 10 microg and IL-2 50 kU, 100 kU or 2 kU (maintaining dosage) at previously designated intervals.

RESULTSBoth the mono-administration of either HSP70 or IL-2 and the compound immunotherapy of HSP70 and IL-2 obviously inhibited the growth of the implanted tumor and prolonged the life span of the mice to different extents. However, long periods of tumor-free survival (over 90 days) were demonstrated only in HSP70 10 micro g group, HSP70 10 microg-IL-2 50 kU group, and HSP70 10 microg-IL-2 100 kU group (40%, 40%, 60% respectively). On the other hand, none of the mice in the rest groups achieved long-term survival. Statistical significance was apparent in comparison with the groups without long period survival (P < 0.025 - 0.05).

CONCLUSIONOur research revealed that tumor-derived HSP70 immunotherapy was much more effective than IL-2 alone. And in compound immunotherapy, HSP70 was the main factor in delaying or eradicating the tumors. The proper combination of HSP70 and IL-2 (10 microg HSP70 and 100 kU IL-2 in this experimental mouse model) clearly enhanced the immunotherapy efficacy which indicated that the specific immunotherapy as a main part of tumor immunotherapy assisted by cytokine immunotherapy would be a promising strategy in cancer treatment.

Animals ; Drug Therapy, Combination ; HSP70 Heat-Shock Proteins ; administration & dosage ; isolation & purification ; therapeutic use ; Interleukin-2 ; administration & dosage ; therapeutic use ; Mice ; Mice, Inbred Strains ; Neoplasms, Experimental ; mortality ; therapy ; Survival Rate ; T-Lymphocytes, Cytotoxic ; immunology ; Tumor Cells, Cultured

OBJECTIVETo study the efficacy and explore the mechanism of the anti-tumor immunity elicited by heat shock protein 70-peptide complexes (HSP70-PC) derived from tumor cells.

METHODSCells culture, flow cytometric analysis, affinity chromatography for protein purification, SDS-PAGE, Western-blotting and animal experiment were used.

RESULTSHSP70-PC immunization rendered protective effect to both naive tumorl-bearing mice. All of the naive mice obtained complete resistance to Hcaf cell attack; 40% of the tumor-bearing mice survived for over 90 days, whereas the mice of control group died within 2 weeks (P < 0.01). CD8+ subset of T lymphocytes in the peripheral blood of immunized mice increased by 12%.

CONCLUSIONHSP70-PC induces anti-tumor immunity via activation of cytotoxic T lymphocytes (CTLs), and it possesses strong tumor vaccine effect. Our research adds more evidence to support the clinical use of HSP70-PC to fight human cancers.

Animals ; CD8 Antigens ; blood ; Cell Membrane ; chemistry ; HSP70 Heat-Shock Proteins ; biosynthesis ; isolation & purification ; therapeutic use ; Liver Neoplasms, Experimental ; chemistry ; drug therapy ; immunology ; pathology ; Mice ; Neoplasm Transplantation ; T-Lymphocytes, Cytotoxic ; immunology ; Tumor Cells, Cultured

Objective: Our aim was to examine the correlation infection of HBV and the formation of cholelithiasis. Methods： Gallbladder bile samples of 38 HBV-infection patients and 35 non-HBV-infection patients were determined. Results： Elevated levels of unconjugated bilirubin(UCB)(P<0.01)and Ga2+(P<0.05),decreased levels of total bile acid (TBA)(P<0.01)and cholesterol (TC)(P<0.01)were found. Conclusio： The changes of bile elements of HBV-infection and cholelithiasis are correlated.

Objective: Our aim was to investigate the alteration of p16 gene in human pancreatic carcinoma. Methods: A total of 66 human pancreatic tissue specimens, comprising 51 with pancreatic carcinomas and 15 normal pancreatic tissue specimens, were examined for homozygous deletion and mutation of p16 gene by using PCR-SSCP method. Results: No mutation and deletion was detected in 15 normal pancreatic tissue samples. Of 51 pancreatic carcinoma specimens, only one was found mutation for p16 gene in PCR-SSCP assay, and the deletion of the p16 gene in 23 samples were confirmed by using PCR, with a 45% p16 gene deletion rate. Conclusion: These data suggest that p16 gene alterations may play a role in the progression of human pancreatic carcinoma.

Objective To evaluate the effect of simple closure operation followed by anti-Helicobacter pylori(Hp)therapy for perforated peptic ulcer. Methods 168 cases of perforated peptic ulcer treated by simple closure operation with or without postoperative medication therapy were followed-up and analyzed. Results 1 year after operation, the recurrence rate of peptic ulcer was 3.8% in the group of standard anti-Hp therapy(S group), and that of gastric and duodenal ulcer in S group was 8.3% and 3.6%,respectively; while it was 62.9%, 66.7% and 58.8% in the group of non-standard Hp therapy (N-S group), and 88.9%, 100% and 80.0% in the group without Hp therapy (NT group).The diffcrence of reccurrence rate of peptic ulcer between S group with N-S group?NT group was significant(P

Objective To investigate the cytological pattern and distribution in nonfunctioning pancreatic endocrine tumors.Methods Using labeled streptavidin-biotin (LSAB), immunohistochemical staining for insulin, glucagon, somatostatin, pancreatic polypeptide and gastrin was performed on 30 nonfunctioning pancreatic endocrine tumors from 30 patients. The cellular composition and anatomic distribution in these tumors were analyzed.Results Of 30 tumor tissues, 22 (73.3%) were found to contain cells immunoreactive to 1-4 kinds of peptide hormones; 17 (56.7%) showed positive staining for more than one peptide and up to 4 peptides; and 8 (26.7%) showed negative immunoreaction to all antiserum applied. No tumor was found to contain immunoreactive gastrin. Among 17 multihormonal tumors, 4 contained 2 kinds of peptide hormones, 8 had 3 kinds, and 5 harbored 4 kinds of peptide hormones. In addition, the difference in the number and type of positive endocrine cells between the tumors arising from the head of the pancreas and those arising from the body and tail of the pancreas were statistically significant (P<0.05).Conclusions Immunohistochemically, the high positive rate to peptide hormones suggests that the nonfunctioning pancreatic endocrine tumors are actually not nonfunctioning; they are asymptomatic pancreatic endocrine tumors. Moreover, an uneven distribution of positive endocrine cells in the nonfunctioning pancreas endocrine tumors within the pancreas was identified.

Objective To investigate the calcification of thyroid nodule detected on ultrasound and the relation with benign and malignant disease.Methods Data of 107 cases of malignant and 703 cases of benign(thyroid) nodules examined by high-resolution colored ultrasonography preoperatively and pathological diagnosis by paraffin embedded slides postoperatively were collected from our hospital over a period of 3 years.The(percentages) of calcification and fine stippled psammomatous(FSP) calcification in benign and malignant(nodular) disease,in different sexes and different age groups were retrospectively reviewed.Results The(incidences) of calcification and FSP calcification were significantly higher in malignant group(63.55%,(29.02%),respectively) than in benign group(P0.05).Calcification rate showed no(significant) difference between different age groups(0.05);however,there was a significant difference of the FSP calcification rate between different age groups(P

Objective: To investigate the successive fluctuation of some Thl type cytokines in the peripheral blood of tumor-bearing mice treated with tumor-derived heat shock protein 70 ( HSP70) , explore the mechanism of HSP 70 in breaking through the tumor-immunity tolerance of the organism with tumor-burden and in inducing effective anti-tumor immune response, and provide valuable reference for tumor-derived HSP70 administration in treating human cancer. Methods : Cell culture, techniques for protein extraction and purification, SDS-PAGE, Western-blot, capillary electrophoresis, ELISA technique and animal experiment were applied. Results: HSP70 could result in apparent tumor-inhibitory effect and upregulate some Thl type cytokines(IL-2, TNF-?, and IFN-?) in the peripheral blood of the treated mice gradually to the frequency of HSP70 administration, and showed no reduction trend in two weeks after the final treatment. Statistically significant difference was observed contrasted with those of the control group (P