Objective To clarify the involvement of the free fatty acids(FFA)and lipid peroxida-tion in rat pancreatic tissue during the development of pancreatic injuries inducecd by long-term high-fat diet.Mehtods The male Sprague-Dawley rats (n=72) were randomized into 6 groups (n=12).One group (group control) received standard chow for 18 weeks, the other five groups (group HFD) were fed with a high-fat diet respectively for 2, 4, 6, 10 and 18 weeks.Serum TG and TCH, the his-topathological changes, pancreatic malondialdehyde (MDA) content, superoxide dismutase (SOD) ac-tivity and the concentration of free fatty acids in pancreatic tissues were examined.Pancreatic fibrosis was assessed using Sirius Red staining.The expression of desmin, a smooth muscle actin (α-SMA), platelet-derived growth factor receptor type β (PDGFRβ) and transforming growth factor131 (TGFβ1) was determined with immunohistochemistry.Results Pancreatic MDA content, the number of desmin and α-SMA positive cells were significantly increased in all the HFD groups.The FFA content, PDGFRβ, and TGFβ1 in pancreatic tissues increased in rats of 2, 4 and 6 week HFD groups accompa-nied with typical histological alternations including edema, capillary vessels hyperplasia, and focal aci-nar degeneration, vaculation of acinar and islet cells.In 6, 10 and 18 weeks HFD groups, the lesions had progressed and acinar cell atrophy, fatty replacement, deposition of hemosiderin, and interstitial collagen deposition were observed.Conclusions The increased amounts of FFA and lipid peroxidant in pancreatic tissues are associated with pancreatic cell injuryies and synthesis of collagen by activated PSCs during the chronic high-fat diet intake.

ObjectiveTo explore the sensitivity of cancer stem cells to chemotherapy in human pancreatic carcinoma.MethodsPANC-1 ceils were cultured in an incubator filled with 5％ CO2 at a temperature of 37℃,and were labeled with Hoechst 33342.The SP analysis and sorting were performed using a FACSVantage SE.RT-PCR was performed to detect the expression of human CD133 ABCG2 and Notch1.SP and non-SP cells from the PANC-1 cell line were treated with 5-fluorouracil (5-FU; 1,10,or 100 μg/ml) or gemcitabine (10,100,or 1000μg/ml),and the cell viability was determined using the MTT assay.The sensitivity of sorted tumor cells to chemotherapeutics was determined in NOD/SCID mice model.ResultsSP cells were found to have higher drug-resistance both in vivo and in vitro and higher levels of mRNA expression for CD133,ABCG2 and Notch1,when compared to non-SP ceils.The xenografted tumors derived from injected SP cells and treated with gemcitabine had more CD133± cells than the untreated group.ConclusionsThe SP of PANC-1 pancreatic carcinoma cells are enriched with highly gemcitabine-resistant CSCs and determine the carcinogenesis of the PANC-1 pancreatic carcinoma cells.

Objective To observe the expression of connective tissue growth factor (CTGF) in pancreas, and discuss its significance. Methods The pancreatic fibrosis model was induced by high fat diets. The rats were sacrificed 16 weeks later, and the pancreatic tissue was harvested for routine pathologic examinations. Pancreatic collagen fibrosis I was determined by HE and Sirius red staining;α-SMA and CTGF expression were detected by immunohistochemistry. Results After pancreatic fibrosis, pancreatic lobules and acinar atrophy was observed, lobules gap was widened, interstitial fibrous tissue was significantly proliferated, the synthesis of pancreatic collagen fibrosis I was significantly increased when compared with normal pancreas ( 1500.2 + 255.8 vs. 57.4 ± 23.2, P ＜ 0. 01 ), the expression of α-SMA was significantly increased when compared with normal pancreas( 1500.2 + 255.8 vs. 57.4 + 23.2, P ＜ 0. 01 ), and the expression of CTGF was significantly increased when compared with normal pancreas (2950.5 ± 431.9 vs. 382.2 + 190.8, P ＜0.01 ), and there were abundant activated PSCs. Conclusions CTGF participated in the regulation of pancreatic fibrosis development; the function of CTGF was closely related to PSCs activation.

ObjectiveTo investigate the morphological and ultrastructural characteristics of pancreatic stellate cells (PSCs) in a chronic pancreatitis murine model.MethodsSprague-Dawley rats were divided into experimental group (n=12) and control group (n=12).The experimental group was fed with a high-fat diet (HFD) for 16 weeks whereas the control group received regular rat feed.Pathologic changes in the pancreas were observed by several morphological studies including a Sirius Red staining method,as well as immunostaining for desmin and a smooth muscle actin (a-SMA).TEM and immunoelectron microscopy were performed to study the ultrastructure of the PSCs.Results The number of desmin and α-SMA positive cells significantly increased in the HFD groups compared to the control group.The quiescent and activated PSC cells both coexisted in the pancreas in the HFD group.ConclusionsThe morphology and ultrastructural study of pancreatic stellate cells may be helpful for further functional studies on chronic pancreatic fibrosis.

Objective To observe the influence on the sensitivity of pancreatic cancer cell line BxPC-3 to gemcitabine of silencing PAUF gene.Methods BxPC-3 cells,which overexpress PAUF,was stably transfected with PAUF-shCtrl and PAUF-shRNA to establish BxPC-3_shCtrl and BxPC-3_shPAUF cells as control and experiment group.Then the mRNA and protein expression level of PAUF in these two cell lines were detected by RT-PCR and western blot,respectively.The growth inhibition rates of these two cell lines treated with different concentrations of gemcitabine (0,3.1,6.25,12.5,25,50,100,200 nmol/L) were detected by MTT.Apoptosis rates in the cells treated with different concentrations of gemcitabine (0,75,100 nmol/L) were then observed by flow cytometry.Results The relative PAUF mRNA expression level in BxPC-3_shCtrl and BxPC-3 cells were 1.00 ± 0.06 and 0.83 ± 0.07,which were significantly high er than that in BxPC-3_shPAUF cells (0.25 ± 0.02;both P ＜ 0.05).The relative PAUF protein expression level in BxPC-3_shCtrl and BxPC-3 cells were 0.89 ± 0.07 and 0.95 ± 0.04,which were significantly high er than that in BxPC-3_shPAUF cells (0.31 ± 0.03;both P ＜ 0.05).The IC50 value of gemcitabine to BxPC-3_shCtrl cell was (22.88 ± 2.43) nmol/L,which was significantly higher than that of BxPC-3_shPAUF cells [(1.06 ± 0.02) nmol/L;P ＜ 0.05];apoptosis rate of BxPC-3_shPAUF cells treated by gemcitabine increased faster than that of BxPC-3_shCtrl cells.Conclusion PAUF silencing could greatly enhance the sensitivity of BxPC-3 cells to gemcitabine.

Objective To explore the function of Notch-3 in epithelial-mesenchymal transition(EMT) and prognosis of pancreatic cancer patients.Methods Patients who received radical resection for pancreatic cancer in our hospital between January 2004 and October 2012 were included in this study.Immunohistochemical staining was performed with Notch-3,E-cadherin and Vimentin antibodies.Imaging pro plus 3.0 was used for analyzing the staining intensity.Survival analysis was performed using Kaplan-Meier method.Results Sixty-seven patients were included.Low expression of E-cadherin was detected in 61.2％ (41/67) of patients,while high expression of Vimentin and Notch-3 was found in 65.6％ (44/67) and 59.7％ (40/67),respectively.Notch-3 expression was proportional to Vimentin expression (R2 =0.872,P ＜ O.05),while inversely proportional to E-cadherin expression (R2 =0.570,P ＜ 0.05).Median overall survival time in high expression group of E-cadherin,Vimentin and Notch-3 was (25.2 ± 2.3) months,(14.8 ±0.9) months and (15.8 ±0.8) months.While in low expression group,the median overall survival time was (14.3 ± 1.0) months,(25.5 ± 2.4) months and (25.1 ± 2.9) months,respectively.There were significant differences between these two groups (all P ＜ 0.05).Conclusions Notch-3 expression was associated with EMT process in pancreatic cancer patients.Low expression of E-cadherin and high expression of Vimentin and Notch-3 predicated poor prognosis of pancreatic cancer.

Abstract Allergic diseases can occur in all systems of the body, covering the whole life cycle, from children to adults and to old age, can be lifelong onset and even fatal in severe cases. Children account for the largest proportion of the victims of allergic disease, Children s allergies start from scratch, ranging from mild to severe, from less to more, from single to multiple systems and systemic performance, so the prevention and treatment of allergic diseases in children is of great importance, which can not only prevent high risk allergic conditions from developing into allergic diseases, but also further block the process of allergy. At present, there is no consensus on the management system of allergic children in kindergartens and primary schools. The "Consensus on Allergy Management and Prevention in Kindergartens and Primary Schools", which includes the organizational structure, system construction and management of allergic children, provides evidence informed recommendations for the long term comprehensive management of allergic children in kindergartens and primary schools, and provides a basis for the establishment of the prevention system for allergic children.