Objective To explore the effects of nicorandil on cardiac function and clinical outcomes in patients with acute myocardial infarction (AMI) undergoing percutaneous coronary intervention (PCI). Method Sixty-six patients with AMI were randomized into a control group and nicorandil group (n = 33 for each group). In the nicorandil group, nicorandil (4 mg as a bolus injection followed by constant infusion at 8 mg/hour for 24 hours) was administered immediately after admission. Reactive oxygen species (ROS) formation was assessed by measuring urinary excretion of 8-epi-prostaglandin F2α (PGF2α) and compared between the two groups; cardiac function and cardiac events were also compared. Results Urinary 8-epi-PGF2αexcretion was increased 2-fold at 60 to 90 minutes after PCI in the control group, whereas it was unchanged in the nicorandil group (P < 0.001). Left ventricular ejection fraction and cardiac index immediately after PCI and at 6 months were greater in the nicorandil group than in the control group(P < 0.05). Rates of total inhospital cardiac events and rehospitalization were lower in the nicorandil group than in the control group (P<0.05). Conclusions Nicorandil improves cardiac function and clinical outcomes in patients with AMI undergoing primary percutaneous coronary intervention. Suppression of ROS formation may be involved in the potential mechanism.

Objective To observe the changes of zymophagy during experimental acute pancreatitis (AP) induced by caerulein.Methods Pancreatic acinar cell line AR42J cells were cultured in 6-well plates till 90％ confluent and then divided into AP group and control group.Caerulein (1 × 10-8 mol/L) was added into AP group to establish AP cell model,and 1640 cell culture medium was added into control group.After caerulein treated for one,four,six,eight,12 and 24 hours,cells and cell culture supernatant were collected.The levels of cytokine interleukin (IL)-1,tumor necrosis factor (TNF)α,trypsinogen activation (TAP) and amylase were measured with enzyme-linked immunosorbent assay (ELISA) method.The expression of LC3 and Beclin1 at mRNA of each group were detected by reverse transcription-polymerase chain reaction (RT-PCR).The LC3B protein level of each group were detected by Western blotting.The changes of autophagosome and zymophagosome were observed by transmission electron microscopy.The difference between AP group and control group was analyzed by analysis of variance.Results The level of IL-1,TNFα,amylase and TAP in cell culture supernatant of control group was (18.83±7.10) pg/mL,(14.20±3.79) pg/mL,(10.03±2.85) U/L and (39.48±8.62) pg/mL,respectively.Those of AP group significantly increased at first hour ((62.13±11.25) pg/mL,F=3.32,P＜0.01 ; (30.98±7.11) pg/mL,F=3.05,P＜0.05; (25.06±6.82) U/L,F=2.90,P＜0.05 and (128.51± 18.30) pg/mL),F=2.62,P＜0.01,at fourth or sixth hour reached peak (IL-1 at fourth hour:(71.96± 15.82) pg/mL,F=7.25,P＜0.01;TNFα at sixth hour:(39.92±8.94) pg/mL,F=4.93,P＜0.05; amylase at fourth hour:(28.83 ± 8.31) U/L,F=2.06,P＜0.05; TAP at fourth hour:(146.29± 29.36) pg/mL,F=0.14,P＜0.01) and then gradually decreased.At fourth and sixth hour,the expression of LC3 at mRNA level in AP group was 3.18±0.82,1.71±0.14,respectively,while the expression of Beclin-1 rnRNA at first,fourth hour was 2.44±0.34 and 4.13±0.30,all of them were significantly increased compared with those of control group (0.21±0.04 and 0.30±0.08,LC3 mRNA F=0.79、0.06; Beclin mRNA F=2.31、0.36,all P＜ 0.05).There were no significant differences at other time points.The numbers of autophagosome and zymophagosome of AP group were significantly higher than those of control group under transmission electron microscopy.Conclusion Zymophagy occurred during AP cell model induced by caerulein,which suggested that zymophagy might involve in the mechanism of AP.

Background:Damage of intestinal mucosal barrier is a key factor in the development and progress of acute pancreatitis(AP),and is closely related with the prognosis of the disease. Aims:To investigate the protective effect and possible mechanism of mucoprotective agent teprenone on intestinal mucosal barrier in rats with experimental AP. Methods:Forty-five adult male Sprague-Dawley rats were randomly divided into normal control group(n = 5),AP model group(n = 20)and teprenone treated group(n = 20). AP model was established by subcutaneous injection of cerulein at abdominal wall. Rats in treated group were intervened with teprenone intragastrically before and after model establishment. ELISA was used for measurement of serum interleukin-1(IL-1),IL-6,tumor necrosis factor-α(TNF-α)and amylase;histopathological and ultrastructural changes of small intestinal mucosa were observed by light microscope and transmission electron microscope;Western blotting was used to detect the expressions of tight junction protein occludin and ZO-1. Results:Serum levels of IL-1,IL-6,TNF-α and amylase in AP model group were significantly higher than those in normal control group(P < 0. 05),accompanied by necrosis and exfoliation of small intestinal villus,widening of intercellular tight junctions and downregulation of occludin and ZO-1 expression. While in teprenone treated group,serum levels of proinflammatory cytokines and amylase were significantly decreased as compared with AP model group(P < 0. 05),the villus of small intestine remained intact,and dense tight junctions were observed. Expressions of occludin and ZO-1 in teprenone treated group were upregulated. Conclusions:Teprenone may protect against intestinal mucosal barrier injury in AP model rats by upregulating tight junction protein expression.

Objective To explore the feasibility of establishing porcine model for training of endoscopic ultrasound (EUS) guided celiac plexus paracentesis.Methods A total of 6 healthy pigs were sedated with an intramuscular injection of Ketamin at 10 mg/kg,followed by intravenous injection of 3% pentobarbital at 0.8 ml/kg.EUS was then performed and empty seeds were implanted into celiac plexus.Enhanced CT scan was performed to confirm the location of the implanted seeds.Results No animal died after the procedure.All seeds were accurately distributed on both sides of the celiac trunk except in one pig the seed was found in stomach by CT scan and was re-implanted another day.Conclusion Pigs are similar to human in anatomic structure and they can be excellent models for beginner endoscopy physicians to acquire the skill of EUS guided celiac plexus paracentesis.

Objective To evaluate the influence of brachytherapy with 125Ⅰ seeds on celiac ganglia in porcine models. Methods Twelve pigs were randomly assigned into 3 groups to accept celiac plexus block by bilateral injection with 2 non-radioactive seeds in group A (n = 4), 0.4 mCi seeds in Group B (n = 4) and 0.8 mCi seeds in Group C (n = 4), respectively. Prophylactic antibiotics were administered postoperatively. Enhanced CT and three-dimensional reconstruction of blood vessels were performed to confirm the proper placement of the seeds. The animals were sacrificed 14 days and 60 days after the procedure, and TUNEL assay was employed to study neuron apoptosis in ediac ganglia. Results The procedure was succossfully performed in,10 pigs(83.3%)and failed in two others. The rescue procedure was performed on the day after and succeeded in both pigs. Apoptosis of neurons significantly increased in brachytherapy groups than in the control group and it was positively correlated with dose and time of radiation. Conclusion Brachytherapy can cause apoptosis of neurons of celiac ganglion, which could be the basis of its clinical application.

ObjectiveTo assess the safety of celiac plexus brachytherapy with 125Ⅰseeds placementguided by endoscopic ultrasound in porcine model, and to evaluate its effect on surrounding vessels and organs.MethodsFourteen pigs were randomly divided into 4 groups to accept celiac plexus block by bilateral injection with 2 non-radioactive seeds in group A (n=4),0.4 mCi seeds in Group B (n=4),0.8 mCi seeds in Group C (n=4) and one lateral injection with 5 ml dehydrated alcohol,respectively.Abdominal X-ray,Enhanced CT and three-dimensional reconstruction of blood vessels were performed to confirm the proper placement of the seeds in group A,B and C.Routine blood test,liver and renal function,serum amylase and CD4+/CD8+ratio were examined preoperatively and at the end of the follow-up in all groups.Animals were euthanized in batch to observe the position of the implanted seeds.Tissues and organs around the seeds were dissected for pathological examination.ResultsThe procedure succeeded in 12 pigs (85.7%)and failed in two others (1 in group A,and 1 in C) due to inappropriate position.Rescue procedures were performed on another day and succeeded.No significant difference was found in routine blood test,liver and renal func-tion,serum amylase and CD4+/CD8+ratio,except WBC elevation and small abcesses were found 7 days after the procedure in one pig of group C.In the radiated area, there was degeneration,necrosis and mild in-flammation at the outer membrane of blood vessels,with fibrosis around,which was positively correlated with the radiation dosage and duration.There was no change at the muscular layer and the innermost membrane of blood vessels,and no thrombosis was found.In group D,the celiac trunk became slightly brown and wider with obvious hemorrhage,necrosis and infiltration of inflammatory cells in the outer membrane of blood vessles and connective tissues around.ConclusionBrachytherapy has little negative effect on the organs a-round. It does not harm immunnity but induces lesions in blood vessels. Compared with dehydrated alcohol,this negative effect is limited.Therefore,celiac plexus brachytherapy with125Ⅰseeds guided by endoscopic ultrasound is safe.

Objective To investigate the clinical benefit response (CBR) in treating the unresectable pancreatic carcinoma by applying the EUS guided iodine-125 seed implantation combined with chemotherapy of gemeitabine and comparing chemotherapy of gemcitabine alone. Methods Forty-one patients with unresectable pancreatic carcinoma were randomly divided into two groups, one group (Group A) included 21 cases which underwent EUS-guided iodine-125 seed implantation combined with gemcitabine chemotherapy, the rest 20 cases (Group B) were treated with gemcitabine chemotherapy alone. EUS-guided iodine-125 seed implantation were carried according to the treatment plan system (TPS), following chemotherapy after 1 week. Gemcitabine was administered at the dose of 1 000 mg/m2, through intravenous administration once a week for 3 consecutive weeks every 4 weeks. CBR was assessed. Results CBR of Group A was 57.1% and median time to CBR was 1 week and median duration of CBR was 21 weeks, while CBR of Group B was 25%, and median time to CBR was 4 weeks and median duration of CBR was 15 weeks (P<0.01). Conclusions EUS-guided iodine-125 seed implantation combined with chemotherapy of gemcitabine was superior to gemcitabine chemotherapy alone in the term of CBR in patients with unresectable pancreatic carcinoma.

Objective To investigate the changes and significance of autophagy in rats with experimental acute necrosis pancreatitis (ANP).Methods According to method of random number,18 rats were randomly divided into control group,ANP group,ANP+rapamycin (RAP) group.The ANP rat model was established by intraperitoneal injection of 20％ L-arginine.The rats of ANP+RAP group were intraperitoneal injected with RAP 1.2 mg/kg at 30 minutes before modeling.The rats of control group were intraperitoneal injected with 0.9％ NaCl solution.The blood was drawed from the hearts nine hours after modeling for subsequent experiments.Serum levels of trypsinogen activation peptide (TAP),interleukin (IL-1),IL-6 and tumor necrosis factor (TNF) α were measured with enzyme-linked immunosorbent assay.The pancreatic tissues were pathologically scored.Autophagy-related structures in rat pancreatic acinar cells were observed by transmition electron microscopy.The expression of autophagy marker microtuble assciated protein 1 light chain 3 (LC3)-Ⅱ and Beclin-1 at mRNA and protein level were measured by quantitative real-time polymerase chain reaction (qRT-PCR),Western bloting and immunohistochemistry.The single factor analysis of variance was used for mean comparison among groups.Results A rat model of ANP was successfully established.Histopathological score of pancreas acinar cell necrosis of ANP+RAP group (2.19±1.38) was higher than that of ANP group (0.97±0.68),and the difference was statistically significant(F=33.75,P＜0.05).The results of Western blotting indicated that the protein expression of LC3-Ⅱ and Beclin-1 in ANP group (35.25±2.68 and 49.40±5.28)were higher than those in control group (1.54±0.16 and 0.78±0.06),furthermore the expressions in ANP+RAP group(123.53±3.21 and 76.41±3.80) were higher than those in ANP group,and the differences were statistically significant(F=2 045.54,326.87,both P＜0.01).Immunohistochemistry results also indicated that the LC3Ⅱ and Beclin-1 expression at protein level of ANP+RAP group (7 570.63±4 357.67 and 3 418.09±2 035.78) were higher than those of ANP group (1 926.53±1 414.44 and 536.11±403.10),and the differences were statistically significant (F=39.83,41.58,both P＜0.01).The expression of Beclin-1 at mRNA level of ANP group (107.12±29.10) was statistically higher than that of control group(7.01 ±3.39),and the difference was statistically significant (F=3.61,P＜0.05),but the expression of ANP+RAP group (97.63 ± 65.38)was no significant difference compared with ANP group.However,the expression of LC3-Ⅱ at mRNA level of ANP+ RAP group (4.37 ± 1.67) was statistically higher than that of ANP group (1.76 ± 1.59),and the difference was statistically significant(F=16.10,P＜0.05),but the expression of ANP group was no significant difference compared with control group (1.51 ±0.95).The result of electron microscopy showed that autophagy related structures increased in ANP group compared with that of control group,which of ANP+RAP group was more.The serum levels of TAP,IL-1 and IL-6 of ANP + RAP group were (36.47 ± 1.71) pmol/L,(122.88± 26.67) pg/mL and (107.39±13.95) pg/mL,which were all higher than those of ANP group ((25.63 ± 6.05) pmol/L,(98.06 ±9.29) pg/mL and (86.16± 7.20) pg/mL),and the differences were statistically significant (F=116.71,50.45,79.67; all P＜0.01).There was no significant difference in TNFα between ANP+ RAP group ((140.80±60.82) pg/mL) and ANP group ((105.23±6.95) pg/mL,F=14.76,P＞0.05).Conclusions Autophagy increased in rats with ANP.Promoting autophagy could significantly activate trypsinogen,aggravate pancreatic injury and increase inflammation reaction,which indicated that autophagy might involve in the pathogenesis of ANP through trypsinogen activation.

Objective To establish a stable AR42J cell line expressing EGFP LC3.Methods The EGFP LC3 overexpressed Lentivirus was constructed and transfected into pancreatic acinar cells (AR42J) of rats.The rats with Lentiviral EGFP transfection were treated as negative control.The transfection efficiency was detected by inverted fluorescence microscope and flow cytometry.The EGFP LC3 protein expression in the stable cell lines were analyzed by Western blot.The cells were treated with thapsigargin to establish endoplasmic reticulum stress model,and the LC3,PERK protein expressions were detected by Western blot.Results The transfection efficiency of Lentiviral EGFP LC3 of AR42J cell was ＞ 85％,which could achieve stable passage.The expression of LC3 mRNA of AR42J cells transfected with Lentiviral EGFP LC3 was 9.14 ±0.32 folds higher than that of negative control,which had no expression of LC3 protein,only EGFP expression.However,compared with non-transfection group,the LC3 mRNA expression in EGFP group was not significantly different.Conclusions A pancreatic acinar cell line (AR42J) of rat stably expressing EGFP LC3 protein is successfully constructed.And it may provide a new model for further research of the relationship between acute pancreatitis and autophagy.

Objective To evaluate the safety and efficacy of direct celiac ganglion irradiation with 125I seeds for pain relief secondary to advanced pancreatic carcinoma (PC).Methods This study enrolled 23 consecutive patients who had moderate to severe pain resulting from advanced PC.All patients underwent EUS-guided direct celiac ganglion irradiation with 125I seeds.Follow-up was conducted at least once weekly until death.Blood parameters,Visual Analog Scale (VAS) score,mean analgesic consumption,and complications were evaluated during follow-up.Results All patients successfully underwent implantation at one attempt.The mean number of seeds implanted in the celiac ganglion per patient was 4 (range 2-6).Immediately after the procedure,pain relief and analgesic consumption showed no significant changes compared with preoperative values.Six patients (26％) reported pain exacerbation.Two weeks later,the VAS score and mean analgesic consumption were significantly less than preoperative values.No procedure-related deaths or major complications occurred.Conclusion EUS-guided direct celiac ganglion irradiation with 125I seeds can reduce the VAS score and analgesic drug consumption in patients with unresectable PC.