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To explore the effect of DAPT, a gamma-secretase inhibitor on the proliferation of pancreatic cancer cells and its underlying mechanism.Three human pancreatic cancer cell lines as AsPC-1, BxPC-3 and MIAPaCa-2 were employed for in vitro experiments. Cell proliferation was tested by CCK-8 and cell apoptosis was examined by Annexin V/PI staining after the treatment of various concentrations of DAPT at different time.DAPT of 100 micromol/L significantly inhibited the proliferation of these 3 cell lines from day 3 to 5. DAPT of 50 micromol/L inhibited the proliferation of BxPC-3 and MIAPaCa-2 cells from day 3 to 5, whereas the effect of inhibition was only observed at day 5 in AsPC-I cell. DAPT of 10 micromol/L inhibited the proliferation of BxPC-3 and MIAPaCa-2 cells at day 5, whereas no effect was found in AsPC-1. Flow cytometry test showed that cell apoptosis rate was higher in DAPT treatment groups compared with control group.DAPT could suppress the proliferation of pancreatic cancer cell, with the mechanism of promoting cell apoptosis.

To investigate the association between Interleukin-10 (IL-10) promoter polymorphism and the gastric cancer risk in Chinese Han patients.DNA was extracted from blood samples of gastric cancer patients (n = 75) and controls (n = 75). IL-10 -1082 promoter polymorphism in both patient and control group (three genotypes distribution: AA, AG and GG) was identified by PCR-RFLP and its relationship with gastric cancer risk, clinic and pathologic features was also analyzed.Patients with gastric cancer had a significantly lower frequency of AA (OR = 0.43, 95% CI = 0.20, 0.92; P = 0.03) than controls. Patients with proximal gastric cancer had a significantly higher frequency of GG (OR = 3.06, 95% CI = 1.12, 8.36; P = 0.03) than those with distant gastric cancer. Patients with advanced (stage II/IV) gastric cancer had a significantly higher frequency of AA (OR = 5.09, 95% CI = 1.05, 24.70; P = 0.04) than those with early (stage I /IV) gastric cancer. When stratified by the Lauren's classification, histological differentiation of gastric cancer, no statistically significant results was observed.This study suggests that the IL-1 1082 promoter polymorphism may be associated with gastric cancer in Chinese Han patients, and the difference in genotype distribution may be associated with the location and stage of gastric cancer.

To investigate the optimal condition of bromodeoxyuridine (BrdU) labeling for bone marrow mesenchymal stem cells (BMSCs) in vitro and the feasibility of in vivo tracing of BrdU-labeling BMSCs.BMSCs were isolated from Wistar rats and were in vitro routinely cultured. The third passage BMSCs was used for identification of special surface antigens by immunohistochemical methods. The purified BMSCs were incubated with BrdU at different concentrations for different incubating time to investigate optimal BrdU concentration and incubating time for cell labeling. The cell labeling index of BrdU was calculated with immunohistochemical analysis. BMSCs labeled BrdU were injected into damaged gastric mucosa of rats by micro injector. The colonization of BMSCs labeled BrdU in gastric mucosa was viewed.After purification and proliferation, the primary cultured BMSCs were uniformly long spindle-shapped form and formed cell colony, which showed the characteristics of stem cell. Immunocytochemistry showed BMSCs were positive for CD44 and CD90, while negative for CD14, CD45. The labeling rate of BrdU increased with the labeling time lasting and reached its height at 48 h. After incubating 48 and 72 hours, the labeling rate of BrdU with a concentration of 10 micromol/L was higher than that of BrdU with a concentration of 5 micromol/L (P0.05) and similar with that of BrdU with a concentration of 15 micromol/L (P0.05). In addition, the BrdU labeling could be detected after five consecutive passages and the labeling time could keep 21 d. The pathological observation demonstrated that BrdU-labeled BMSCs could colonize the damaged gastric mucosa with normal morphologic characteristics during observation period.BrdU labeling might be a feasible method for dynamic observation of the migration, growth and differentiation of migrating BMSCs in colonizing sites.

To explore a suitable method for isolation, purification and multiplication of bone marrow mesenchymal stem cells (BMSCs).Density gradient centrifugation and adherence separation methods were applied for isolation of BMSCs from Wistar rats. The cells were cultured and proliferated in culture medium containing calf serum (CS), fetal bovine serum (FBS), free of serum or different volume fraction of FBS. The characteristic and the morphology of BMSCs were observed under inverted microscope every day. The growth curves were draw and the surface antigen of BMSCs were detected by immunocytochemistry technique. The microstructure was observed by transmission electron microscope (TEM).The pure primary cells can be procured by density gradient centrifugation. But the primary cells cultured by adherence separation methods demonstrated higher cytoactive, more rapid proliferation, earlier colony confluence and shorter time for passage than that cultured by density gradient centrifugation method. The cells by adherence separation methods were essentially purified at passage 4. Both CS and FBS can promote the growth and proliferation of BMSCs, but the colony forming efficacy of cells (46.50%) cultured in medium containing 0.12 volume fraction FBS was the highest. The cells surface markers CD44, CD90 were positive and CD14, CD45 were negative. BMSCs were observed by TEM and possessed the characteristic of stem cells. Conclusion BMSCs with high quality and activity can be obtained with adherence separation by suitable method and culture conditions. L-DMEM medium containing 0.12 volume fraction FBS showed more profitable for the growth and proliferation of BMSCs.

To compare the effects of enteral nutrition (EN) and parenteral nutrition (PN) on gut mucosal barrier function and intestinal epithelial tight junctions in rats after surgical stress.Fifty SD rats with surgical trauma were randomly divided into three groups: total parenteral nutrition (TPN) group and enteral nutrition (EN) group with isocaloric and isonitrogenous nutrition and placebo group. Nutrients were administered via the neck vein and needle jejunostomy for the TPN group. The homogenated tissues of liver, lung, and mesenteric lymph nodes were cultured to determine the bacterial translocations rates. The transmembrane binding proteins (occludin) were measured with immunohistochemistry. The ultrastructure and morphology of intestinal epithelial tight junctions were observed by electron microscope. The feces in cecum were cultured for anaerobic bacterial growth analyses.The EN group had more lactobacteria and bifydobacteria than the TPN group, but not statistical significant. The EN group had greater expression of occludin in the intestines than the TPN group. Furthermore, the intestinal epithelial tight junction and microvilli of the EN group were more intact compared with those of the TPN group. The bacterial translocations rates of liver, lung and mesenteric lymph nodes were significantly lower in the EN group than in the TPN group.Neither EN nor standard PN maintains intestinal membrane barriers. But EN increases the expression of transmember binding proteins, maintains the gut epithelial tight junction, improves intestinal mucosal barrier and reduces gut bacterial translocation.

To analyse the expression of mouse mammary tumor virus-like (MMTV-like) gene sequencesin breast cancer of Chinese women.Using nested PCR that amplify MMTV-like gene sequences, we tested the breast cancer tissues from 131 Chinese patients. The result was analyzed and compared with postoperative pathology and prevalence that had been reported from different populations.MMTV-like gene sequences were amplified in 22 of 131 (16.8%) breast cancer biopsy tissues from Chinese patients, which had no relation to pathology, but had the significant difference from other populations.MMTV-like gene sequences can be found in breast cancer tissue from Chinese women with breast cancer. This supports the contention that these sequences may represent a virus associated with human breast carcinogenesis. The differences in prevalence of MMTV-like gene sequences and breast cancer rates among populations may be related to geographic distribution.

This is a randomized controlled trial to evaluate the efficacy of recombinant human growth hormone (rhGH) in preventing the development or reducing the duration of postoperative fatigue syndrome (POFS).Informed consent was obtained from each prospective subject. A total of 48 consecutive patients undergoing the elective abdominal operations were randomly divided into the rhGH group and the placebo group. The rhGH group was given daily rhGH 0.15 U/kg from day 3 to day 9 after operation. The two groups had similar nutritional intakes. Plasma proteins, fatigue (Christensen score) and body weight were investigated before and after operations. Nitrogen balance was measured from day 3 to day 9 after operation.On postoperative day 10, the fatigue score in rhGH group was significantly lower than that in control group (P0.05); the level of fibronectin in rhGH group was significantly higher than that in control group (P0.05). Furthermore, a positive nitrogen balance was achieved and kept in the rhGH-treated group during the postoperative period from the third day onward, whereas it was not achieved in the control group till day 9 postoperatively. Cumulative nitrogen balance was significantly higher in rhGH group than in control group (P0.05). There was no significant difference between the two groups in weight loss after operation (P0.05).The application of low doses of recombinant human growth hormone can improve positive nitrogen balance and reduce the courses of POFS. The serum fibronectin level is proved to be a reliable biochemical indicator of POFS.

To detect the expression of IGFBP-2 mRNA in gastric cancer tissues and its roles in neoplasia and peritoneal metastasis of gastric cancer.The IGFBP-2 mRNA expression levels in the fresh tissues from normal mucosa, primary cancer and peritoneal metastasis of 35 patients with gastric cancer were determined by the quantitative real time RT-PCR method, and the samples were sliced up and stained with hematoxylin-eosin for pathologyical examination.The levels of IGFBP-2 mRNA expression in gastric primary cancer tissues were significantly higher than those in the tissues from normal mucosa and peritoneal metastasis (P0.05). Yet, IGFBP-2 mRNA expression levels showed no difference between the tissues of normal mucosa and peritoneal metastasis (P0.05).The expression of IGFBP-2 mRNA possibly plays an important role in proliferation, differentiation and peritoneal metastasis of gastric cancer, and it might be useful in a new method for clinical prognostication and treatment of gastric cancer peritoneal metastasis.