Objective To isolate and purify pancreatic ductal epithelial cells in adult rats,and induce differentiation of pancreatic ductal epithelial cells to islets in vitro.Methods By retrograde in- jection of collagenase into biliary-pancreatic tract,pancreatic tissues were digested and different types of pancreatic cells including islets,duct and degranulated aicni cells were separated by means of density gradient centrifugation.Ductal cells were purified by adhering method and identified by immunocyto- chemistry stain of ductal epithelial cells maker antigen(Cytokeratin 19,CK-19).Ductal cells were ex- panded in RMPI 1640 with 10% FBS.About one week when most adherent ceils were of monolayer, the medium was changed to serum-free DMEM/F12 supplemented with keratinocyte growth factor (KGF)to further expand ductal epithelial cells.When ceils reached 80% confluence,nicotinamide and high concentration of glucose were added to promote differentiation of pancreatic ductal epithelial cells.Islets like-structure was stained by Dithizone.Results Irnmunocytochemistry stain of CK-19 re- vealed that most isolated ceils were ductal epithelial ceils.The cultured ductal epithelial cells began to adhere at day 1,reached 80% confluence and cell clones were formed at day 14-21.At day 28,islets- like-structure appeared and was positive for Dithizone staining.Conclusions Ductal epithelial cells of rats can be isolated by means of density gradient centrifugation and purified by adhering method.Duc- tal epithelial cells can differentiate into islets-like-structure in vitro.

ObjectiveTo investigate the effects of insulin on the proliferation and invasion of human pancreatic cancer cells PANC1,and on its HIF-1α,VEGF expression.MethodsPANC1 was pretreated with insulin of different concentrations (0.1,1,10,100 nmol/L).The proliferation of PANC1 was tested by MTTmethod,and transwell assay was used to test the invasion ability of PANC1.HIF-1α,VEGF and PCNA protein expression was assessed by Western blots,and HIF-1α,VEGF mRNA was detected by real-time PCR.Results Insulin could increase the proliferation of PANC1 in a dose-dependent manner (p ＜0.05 ),and increase the expression of HIF-1α,VEGF protein.After 100 nmol/L insulin treatment for4 d,the PCNA protein expression in the insulin group was significantly higher than that in the control group (1.196 ±0.014 vs 1.157 ±0.013,P ＜ 0.05).The cancer cells passed through the chamber in insulin group were much more than that in the control group ( 141.0 ± 2.1 vs 89.0 ± 1.4,P ＜0.05 ).The expression of HIF-1α protein was significantly increased (1.139 ±0.020 vs 0.598 ±0.013,P ＜0.05),while there was no significant change of HIF-1αmRNA expression.Both the expression of VEGF protein and mRNA were significantly increased (1.011 ± 0.023 vs 0.627 ± 0.013 0.970 ± 0.016 vs 0.350 ± 0.01 3,P ＜ 0.05 ).Conclusions High insulin microenvironment could enhance the proliferation and invasion of PANC1 cells by up-regulating the expression of HIF-1α and VEGF.

Objective To investigate the changes of extracellular histones during the course of acute liver failure in mice as well as its therapeutic potential.Methods WT mice (C57BL/6) were randomly (random number) allocated to inducing acute liver failure by lethal doses of GalN/LPS injected i.p.Hepatic function,apoptosis of hepatocytes and histological indexes were measured at different intervals following GalN/LPS challenge.The levels of extracellular histones were determined by using ELISA and Western blot methods.Meanwhile,GalN/LPS-treated mice were administered with anti-histone H3 and antihistone H4 neutralized antibodies,respectively.Results Administration of GalN/LPS to mice caused acute liver failure,characterized by significant elevation of plasma ALT levels and massive hepatocyte apoptosis or necrosis.All mice died within 9-12 hours.The levels of nucleosomes and extracellular histones H3 and H4 were increased considerably in a time-dependent manner.The survival rates in GalN/LPS-treated mice were improved remarkably following administration of anti-histone H3 and H4 neutralized antibodies (P =0.037,P =0.025),likely due to the significant inhibition of TNF-production.Conclusions Extracellular histones are an important mediator implicated in the pathogenesis of acute liver failure.Anti-histones show promising potential in the treatment of acute liver failure,which deserves further investigation in the future.

Objective To examine the expression of protein kinase signaling cascade protein kinase B (Akt)-which plays an im-portant role in a number of key biological functions in cellular processes after spinal cord injury (SCI) in rats ,and its effects on SCI induced motor function defects ,and to provide the molecular mechanism for repairing SCI .Methods SCI was produced by extra-dural contusion using modified Allen′s stall with damage energy .The rats were divided into three groups :sham-operated group (Sham) ,interference group (Int) ,and control group (Con) .Using immunostaining studies ,Western blot analyses and real-time qualitative RT-PCR analyses ,we detected the changes of Akt expression at the protein and mRNA level in spinal cord tissues at 1 , 7 ,and 14 d after surgery ,and evaluated the presence and extent of neurological impairment after SCI by the BBB locomotor rating scale .Results The sham operated groups exhibited low expression of the Akt signaling at the protein and mRNA level ,and the ex-pressions increased following SCI .Compared to the animals in the sham operated groups ,prominently elevated level of Akt signaling was detected in the injured spinal cords of SCI group .LY294002 ,an inhibitor of phosphatidylinositol 3-kinase (PI3K) that initiates Akt phosphorylation ,prominently inhibited the expression of Akt produced by SCI and the spontaneous recovery of motor function after SCI .Conclusion Activated protein kinase signaling cascade Akt might be the important intervention aspects of involving in neuroprotective and repair process after SCI .

This study examined whether insulin-stimulated hypoxia-inducible factor 1alpha (HIF-1alpha) expression plays a crucial role in promoting the proliferative vitality and invasive capability in human pancreatic cancer cells. PANC-1 cells were divided into three groups: Control group, insulin group and insulin+YC-1 (a pharmacological inhibitor of HIF-1alpha) group in terms of different treatments. Cells in the insulin group or insulin+YC-1 group were treated with insulin (0.1, 1, 10 and 100 nmol/L) alone or combined with 3-(5'-hydroxymethyl-2'-furyl)-1-benzyl indazole (YC-1, 0.1, 1, 10 and 100 mumol/L). HIF-1alpha mRNA and protein expression in PANC-1 cells was determined by real-time RT-PCR and Western blotting respectively. Cell proliferation and invasion were measured by using growth curve and invasion assay, respectively. Western blot analysis demonstrated that insulin dose-dependently increased the HIF-1alpha protein expression, and YC-1 could dose-dependently block this effect. However, neither insulin nor YC-1 altered HIF-1alpha mRNA levels in PANC-1 cells. Moreover, insulin could enhance the proliferation and invasion of PANC-1 cells, while YC-1 could weaken this effect. It was concluded that the malignant proliferation and local invasion of pancreatic cancer cells may be related to high-insulin microenvironment. The tumor biological behavior change resulting from high-insulin microenvironment may be associated with the increased expression of HIF-1alpha protein.

Objective To study the way in showing collateral ligaments of knee by MRI and MRI findings of the normal collateral ligaments.Methods MR imaging examinations were performed in 55 healthy volunteers . MR imaging included T1WI and PDWI sequence on the coronal, sagittal and coronal posterior oblique plane, in order to observe the collateral ligaments, and to measure the medial collateral ligament and the lateral collateral ligament dimension, which were compared with the results of the cadaver published in the related anatomic literatures.Results (1)On sagittal MRI, the angle between the line of medial collateral ligament and the tibia long axis was 0.55°±0.25°,the angle between the line of lateral collateral ligament and the long axis of the fibular neck was 11.47°±1.61)°.(2)On coronal MRI, 96% of the medial collateral ligaments was completely showed on only one slice, at the same time, 82% of the lateral collateral ligaments was completely showed on only one slice .On 11°coronal posterior oblique plane,90% of the lateral collateral ligaments was completely showed on only one slice .(3)The normal medial and lateral collateral ligaments were hypointense string on both T1WI and PDWI coronal MR images , with an average length of 11.53 cm for the medial collateral ligament and 5.31 cm for the lateral collateral ligament.Conclusion Using sagittal plane completely shown the tibia and fibular neck as the standard plane and thickness of 3mm in examining the collateral ligaments of knee by MR imaging can display the medial and the lateral collateral ligament clearly .

Objective To evaluate a duodenum-preserving total pancreatic head resection procedure for the treatment of chronic panereatitis in patients with a pain-inducing enlarged pancreatic head. Methods From January 1999 to December 2006, 35 cases underwent duodenum-preserving total pancreatic head resection procedure without segment resection of the duodenum, as a modified Beger's procedure. Pain scale in the EORTC QLQ-C30 questionnaire was used to estimate the effect of the surgical procedure on pain relief, and oral glucose tolerance test (OGTF) was used to estimate the maintenance of endocrine function. Results For the anastomosis of the distal pancreas and the jejunum, end-to-end invagination anastomosis was performed in 21 cases, end-to-side duct to mucosa anastomosis was performed in 10 cases, and side-to-side duct drainage procedure was performed in 4 cases. Additional T-tube drainage of the common bile duct was adopted in 4 cases for a possible injury of the common bile duct, and anastomosis of the common bile duct and the duodenum was performed in 1 case for common bile duct obstruction. The mean operation time was 286±55 min, and the mean red blood cell (RBC) transfusion was 1.4±1.3 units. The mean hospital stay was 13±4 days. The mortality of the surgical procedure was 0. The overall morbidity was 17%. Pancreatic fistula developed in 1 case, bile leakage in 3 cases, wound disruption in 1 case, intraabdominal bleeding in 1 case, and there was no duodenal fistula. After the surgery, the mean EORTC QLQ-C30 pain scale decreased from 59±27 to 13±21. On follow-up the endocrine function remained stable, and no new case of diabetes was found. Conclusion The duodenum-preserving total pancreatic head resection procedure without segment resection of the duodenum has good postoperative outcomes, and benefits extirpation of inflammatory pancreatic lesions of the head and uncinate process. It is a safe and effective surgical procedure for chronic pancreatitis with an enlarged and painful pancreatic head.

ObjectiveTo investigate the effect of insulin on the expression of hypoxia-inducible factor-1α in human pancreatic cancer cell line ASPC-1.MethodsWe divided ASPC-1 cells into five groups: normoxia; normoxia stimulated with insulin; hypoxia; hypoxia pretreated with different concentration of insulin; hypoxia of different time points pretreated with same concentration of insulin. Real-time PCR was used to test the expression of HIF-1α mRNA. Immunohistochemistry was used to examine the expression of HIF-1α in ASPC-1 of insulin treated cancer cells. Western blot was used to determine the expression of HIF-1 α protein in those cells. Transwell was used to test whether insulin could enhance the invasion ability of ASPC-1 pancreatic cancer cells.ResultsInsulin promotes HIF-1α protein expression. ASPC-1 cells expressed low levels of HIF-1α protein under normoxic condition.After stimulated with insulin, the expression of HIF-1 α protein significantly increased (P ＜ 0. 05 ). After treated with hypoxia, the expression of HIF-1α protein also increased(P ＜ 0. 05 ). Low concentrations of insulin didn't increase the expression of HIF-1α under hypoxic environment ( P ＞ 0. 05 ), while high concentration of insulin could increase its expression(P ＜ 0. 05). When ASPC-1 cells pretreated with insulin suffered from hypoxia, the expression of HIF-1α first increased then decreased moderately( P ＜0. 05). Insulin could enhance the invasion ability of pancreatic cancer cells( P ＜ 0. 05 ).ConclusionsInsulin mediates the expression of HIF-1α protein in human pancreatic cancer ASPC-1 cells with the characteristics of dose and time dependency. Insulin could enhance the invasion ability of ASPC-1 cells.

Hyperthermia is an efficient type of cancer treatment in which body tissue is exposed to high temperatures to damage and kill cancer cells. Previous studies have focused on the treatment of tumor, however,it can not substitute for traditional methols. In recent years,new research in shows hyperthermia plays an important role in bone metastasis pain control because of the advantages of width rang,rapid onset and noninvasive, and it is therefore well used in. It is also becoming one of classical methods for bone metastasis from cancer. This article reviews recent research and progress of mechanisms of hyperthermia in relief of bone metastasis pain.
Bone Neoplasms ; physiopathology ; secondary ; therapy ; Humans ; Hyperthermia, Induced ; Pain, Intractable ; therapy

The main treatment strategies for chronic pancreatitis in young patients include therapeutic endoscopic retrograde cholangio-pancreatography (ERCP) intervention and surgical intervention. Therapeutic ERCP intervention is performed much more extensively for its minimally invasive nature, but a part of patients are referred to surgery at last. Historical and follow-up data of 21 young patients with chronic pancreatitis undergoing duodenum-preserving total pancreatic head resection were analyzed to evaluate the outcomes of therapeutic ERCP intervention and surgical intervention in this study. The surgical complications of repeated therapeutic ERCP intervention and surgical intervention were 38% and 19% respectively. During the first therapeutic ERCP intervention to surgical intervention, 2 patients developed diabetes, 5 patients developed steatorrhea, and 5 patients developed pancreatic type B pain. During the follow-up of surgical intervention, 1 new case of diabetes occurred, 1 case of steatorrhea recovered, and 4 cases of pancreatic type B pain were completely relieved. In a part of young patients with chronic pancreatitis, surgical intervention was more effective than therapeutic ERCP intervention on delaying the progression of the disease and relieving the symptoms.