Chronic pancreatitis (CP) is a progressive inflammatory condition characterized by repeated attacks of abdominal pain and the destruction and fibrosis of the pancreatic parenchyma which causes to reduced exocrine and endocrine dysfunctions.Recently,the prevalence of CP has increased around the world.More and more researchers have realized the importance of early and accurate ways of diagnosis.The history of patients and the different pathogenic factors,such as alcohol and biliary obstruction,are important components of the ways.Moreover,differenting clinical symptoms and signs,imaging findings,laboratory examinations are necessary for diagnosis.Biopsy remains the gold standard for diagnosis.Integrating these factors,you can diagnose CP in early stage and treat it effectively as soon as possible.

Objective To investigate the best method of transfecting total RNA extracted from pancreatic cancer MiaPaCa-2 cells into dendrtic cells (DCs). Methods DCs were cultured from peripheral blood mononuclear cells induced by rhGM-CSF, rhIL-4 and TNF-α. Morphology of DCs was observed. Flow cytometry was used to detect the mature DCs specific surface markers:CD40, HLA-DR, CD83, CD86. Mixed lymphocyte (MLR) was used to determine the ability of DCs to stimulate allogeneic T cell proliferation.Liposomal transfection, electroporation method and passive transfection was used to transfect MiaPaCa-2 cell total RNA into DCs, Real time RT-PCR and MTT assay was used to determine the expression of MUC1 mRNA and the survival rate of the RNA transfected DCs. Results The cells acquired showed typical DCs morphology, the positive rate of CD40, HLA DR, CD83 and CD86 were 34.3% ,50.2% ,89.2% and 73.6%,and they showed a strong ability to stimulate allogeneic T cell proliferation. 48 h after transfection with MiaPaCa-2 cells total RNA by using electroporation, the MUC1 mRNA amount (45.39 ± 9.33) in DCs was higher than that of liposomes method (3 1. 68 ± 7.25) and passive transfection method (18.53 ± 3.26) . DCs survival rate was (80.36 ± 2.43)% by using electroporation, which was relatively lower than (91.48 ±5.42) % by using passive transfection method, but higher than (67.44 ± 2.51) % by using liposomes method,and it was stabilized around 80%. Conclusions Transfecting total RNA extracted from pancreatic cancer MiaPaCa-2 cells into DCs with electroporation is efficient and safe.

Objective To investigate the induction of specific anti-tumor immune response by transfected dendritic cells (DCs) with survivin mRNA of human pancreatic cancer, and to provide the experimental evidences for the treatment of human pancreatic cancer with DCs vaccine. Methods DCs were isolated and cultured from peripheral blood mononuclear cells (PBMCs). After being transcripted and amplified, survivin mRNA was transfected into DCs by electroporation. The expression of survivin in DCs at different time points was detected by quantitative real-time PCR. The survival rate of DCs before and after transfection was determined by MTT method. The induction of specific cytotoxic T lymphocyte (CTL) response by survivin mRNA transfected DCs was measured by 51Cr standard cytotoxicity test. The induction of specific CTL activation by survivin mRNA transfected DCs was evaluated through testing released IFN-γ by ELISA method. Results After survivin mRNA transfection for 48h, the expression of survivin mRNA in DCs reached the highest point (46.09±6.57). After transfection, the survival rate of DCs was stabilized around 80%. The DCs transfected with survivin mRNA could effectively induce HLA-A2+ / survivin+ specific CTL immune responses. Stimulated with pancreatic cancer cell line Capan-2 cells or SCL-1 cells as control group, the IFN-γ released in 24 hours by survivin specific CTL were (28.79±5.70) U/ml and (25.12±2.13) U/ml respectively, there was no significant difference (P=0.761). Conclusion The induction of CTLs by DCs transfected with human pancreatic cancer survivin mRNA could produce specific anti-tumor immunity.

Pancreatic encephalopathy (PE)is a serious complication of pancreatitis,with difficulties in early diagnosis and poor prognosis. This article introduces the possible pathogenesis of PE involving pancreatin activation,cytokines,infection,water and electrolyte imbalance, and vitamin deficiency,summarizes the clinical manifestations and laboratory features of PE,and points out that the clinical manifestations of PE lack specificity and there are no reliable biochemical indices or diagnostic criteria.This article also elaborates on the diagnosis and treat-ment strategies for PE and points out that the key to PE treatment is active and effective treatment of the primary disease.Most PE patients are improved with the control of pancreatitis.

Pancreatic cancer is a common malignant neoplasm of the pancreas with an extremely high mortality.Currently,the early diagno-sis of pancreatic cancer is still not ideal.Attention should be paid to some clinical warning symptoms,such as unexplained abdominal and back pain,jaundice,and unexpected diabetes.Additionally,the combined use of CA19-9,CEA,and other tumor markers,the attention to biochemical indicators,the detection of mutation in KAI1 or p53 gene,and the exploration of the value of miRNA in clinical diagnosis are of great significance.On the other hand,ultrasound,CT,MRCP,ERCP,PET-CT,and other imaging methods,as well as effective col-lection of cytology specimens,should be performed.Thus,there is hope for the early diagnosis of pancreatic cancer.

Objective To measure the intra-brace pressure in a new fabricated three-dimensional dynamic brace and the intra-cast pressure in the rigid plaster cast,and compare the differences of pressures between the brace and cast. Methods Twenty healthy adult female volunteers involved in this study,with the mean age of 38.4 years.ranged from 26 to 44 years.The forearms of every subject were applied a traditional Dlaster cast and a new fabricated forearm three-dimensional dynamic brace respectively to immobilize the forearm and hand with the wrist at a neutral position,and the applying order was randomly.The intra-brace and intra-cast pressures were recorded at 8 points by using the Fuji pressure sensitive film with the hand at a position of fist and that of extended fingers.The 8 points were the bakc of hand,styloid process of ulna,dorsum of distal radius,abdomen of middle forearm,palm,metacarpus of distal radius,flexion muscle belly,and extension muscle belly.The pressures were compared between intra-brace and intracast.Results The dynamic brace provided greater holding force at the hand and at the distal orearm than the Dlaster cast,and greater immobilization at the wrist while allowing full finger function and greater support to the forearm during the hand movements.Conclusion The results indicates that the new fabricated dynamic brace has measurableadvantages in the immobilization compared to the plaster cast,and especially it can Drovide a persistent traction between the hand and forearm which is important for preventing displacement of the fractured segments.The uew dynamic brace can provide a new technique and represent a further development in the conservative management.

ObjectiveTo study the change of autophagy of human pancreatic cancer cell MiaPaCa-2 before and after Ad5-KAI1 tranfection,and to investigate the possible mechanism.MethodsThe MiaPaCa-2 cells without KAI1 expression were infected with Ad5-KAI1 with KAI1 target gene,and Ad5-null was used as negative control,and parental cell was used as blank control.The formation of autophagosomes was observed by electromicroscopy.The green fluorescent protein-labeled light chain 3 (LC3) associations with autophagosome membranes was detected by confocal microscopy.PD98059,LY294002 were applied to pre-treat the cells.The expression levels of beclin 1,AKT,ERK,the phosphorylation of AKT and ERK protein and the ratio of LC3-Ⅱ to LC3- Ⅰ were detected by Western blotting.ResultsAfter 100 MOI Ad5-KAI1 infections for 24 h,the rate of cell expressing KAI1 protein reached (84.97 ±8.56)％,number of LC3 increased from 4 to 20; and swelling,degeneration of mitochondria was observed,and bilayer-like structure in cytoplasm was found.The expression of beclinl increased (1.4 ±0.3 ) folds,and the expression of LC3-Ⅱ/LC3- Ⅰ increased (8.00 ±2.78) folds.PI3K blockade LY294002 pretreatment significantly suppressed the phosphorylation of AKT of MiaPaCa-2 (2.756 vs 1.516),but it did not inhibit the increase of ratio of LC3-Ⅱ to LC3- Ⅰ(0.770 vs 1.403).ERK blockade PD98059 pretreatment not only significantly suppressed the phosphorylation of ERK of MiaPaCa-2 ( 1.637 vs 0.403 ),but also inhibit the up-regulation of beclin 1 protein expression ( 2.377 vs 1.150) and increase of ratio of LC3- Ⅱ to LC3- Ⅰ (2.225 vs 0.680).ConclusionsKAI1 can significantly induce autophagy of human pancreatic cell line MiaPaCa-2 through phosphorylation of ERK rather than AKT.

Objective To explore the pathway of KAI1 induced autophagy regulating apoptosis in human pancreatic cancer cell line MiaPaCa-2.Methods There were three groups in the experiment,which were extracellular regulated protein kinases (ERK) phosphorylation inhibitor PD98059 pretreated group,Caspase-3 activation inhibitor VAD-FMK pretreated group and no PD98059 or VAD-FMK pretreated groups.And each group was divided into three sub groups with different treatment,which were adenovirus AD5-null vector infected control group,the human KAI1 gene recombinant adenovirus vector AD5 KAI1 infected group and autophagy inhibitor 3-MA pretreated and AD5-KAI1 infected group.The cell apoptosis was observed by AnnexinV-FITC/PI double staining.Caspase-3 activation level was evaluated by flow cytometry.ERK phosphorylation and poly(ADPribose) polymerase (PARP) cleavage were determined by Western blot.Results After the cancer cells infected with AD5 KAI1,KAI1 protein was expressed and GFP-LC3 green particles increased.Caspase-3 activation,PARP cleavage,ERK phosphorylation and apoptosis increased obviously.After autophagy inhibitor 3-MA pretreated,the percentage of apoptosis increased from (63.0 ± 7.9)％ to (88.0±4.5) ％ and Caspase-3 activation increased from (34.0±2.8) ％ to (44.2±4.0) ％ and PARP cleavage more.The apoptosis induced by 3-MA could be totally inhibited by Caspase-3 activation inhibitor VAD-FMK pretreated but could not be inhibited by ERK phosphorylation inhibitor PD98059.Conclusion KAI1- induced autophagy inhibits apoptosis through the downregulation of Caspase-3activation and PARP cleavage instead of ERK phosphorylation.

Objective To study the effect of suppressing metastasis between KAI1 and ME491 on pancreatic cancer.Methods Normal pancreases and pancreatic cancers were analyzed using Northern blot,in situ hybridization and immunohistochemistry.Results The expression of KAI1and ME491 was higher in pancreatic cancer than that in normal pancreases,KAI1 expression was higher in pancreatic cancer without metastasis than that in pancreatic cancer with metastasis(P0.05).Conclusion The gene of KAI1 may be involved in the metastasis of pancreatic cancer,ME491 gene may have no effect on the metastasis of pancreatic cancer.

ObjectiveTo investigate the clinical features of patients with painless acute pancreatitis ( AP),and to improve the diagnosis accuracy of painless AP.MethodsThe clinical data of 12 patients with painless AP were retrospectively collected from January 2007 to January 2011.Results The mean age of patients with painless AP was 52 years old.Seven (58.3％) patients complained of abdominal distension and discomfort,4 (33.3％) patients complained of nausea and vomiting,abdominal tenderness occurred in 7(58.3％ ) patients.Serum levels of lipase and amylase was increased in 11 (91.7％) and 8 (66.7％)patients,respectively.The diagnostic accuracy of abdominal ultrasound and CT for AP was 58.3％ and 91.7％,respectively.Five (41.7％) patients were diagnosed to have AP upon admission,and 4 patients were misdiagnosed to have non-digestive diseases.There were 7 severe AP patients among the 12 painless AP patients,and the percentage was significantly higher than that in AP patients with pain (65/327,x2 =7.30,P ＜ 0.05).Painless AP patients needed longer hospital stay when compared with that of AP patients with pain [ ( 20.4 ± 9.1 ) d vs.( 12.9 ± 6.2) d,t =2.296,P ＜ 0.05 ) ].ConclusionsThe misdiagnosis rate of painless AP is high and patients with painless AP are in critical situation,and early measurement of serum levels of lipase and amylase,as well as CT scan is important for correct diagnosis.