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Objective To evaluate the effect and mechanism of rivaroxaban, an inhibitor of coagulation factor Ⅹa (FⅩa), on endotoxin-induced injury to human umbilical vein endothelial cells (HUVEC). Methods When cultured HUVEC grow to 80% fusion, they were divided into four groups according to the random number method: blank control group (DMEM medium), lipopolysaccharide (LPS) group (cells were challenged by 100 μg/L LPS for 16 hours), FⅩa+LPS group (cells were challenged by LPS for 16 hours after they were cultured with 100 nmol/L FⅩa for 24 hours), and FⅩa+RIV+LPS group (cells were challenged by LPS for 16 hours after they were cultured with 100 nmol/L FXa and 1 μmol/L rivaroxaban for 24 hours). After each group of cells were challenged with LPS, the cell activity was detected by the cell proliferation and toxicity kit (CCK-8); the cell migration ability was detected by cell scratch experiments;the abilities of cells migration were measured by scratch-wound-healing assay; the apoptosis of cells were evaluated using flow cytometry; the endothelial barrier of cells was assessed by Transwell and Evans blue; the levels of tumor necrosis factor-α(TNF-α), interleukin (IL-1β, IL-6) were detected by the enzyme linked immunosorbent assay (ELISA); the expressions of nuclear factor-κB (NF-κB) and mitogen activated protein kinase (MAPK) signaling pathway were detected by Western Blot. Results Compared with blank control group, the cell viability in LPS group was significantly decreased, and the migration ability, number of apoptotic cells, and barrier permeability of endothelial cells was significantly increased, the levels of TNF-α, IL-1β and IL-6 were significantly increased, and the expressions of phosphorylation of c-Jun N-terminal kinase (p-JNK), phosphorylation of p38MAPK (p-p38MAPK), phosphorylation of transforming growth factor kinase 1 (p-TAK1) and phosphorylation of NF-κBp65 (p-NF-κBp65) were significantly increased. It indicated that LPS could stimulate the inflammatory response of vascular endothelial cells, and had a significant impact on cell activity, apoptosis and function. There was no significant difference in above indexes between FⅩa+LPS group and LPS group, except for the level of IL-6 being higher in FⅩa+LPS group. Compared with FⅩa+LPS group, in FⅩa+RIV+LPS group, the cell activity was significantly increased (A value: 0.42±0.02 vs. 0.33±0.02), and migration ability was significantly decreased (folds: 1.78±0.17 vs. 2.24±0.20), the number of apoptotic cells was significantly decreased [(11.30±0.70)% vs. (21.03±0.19)%], and permeability of monolayers endothelial cells was significantly decreased [(149±12)% vs. (253±15)%], the levels of inflammatory cytokines were significantly decreased [IL-1β(ng/L): 163.2±20.7 vs. 477.8±20.2, IL-6 (ng/L): 69.3±0.5 vs. 238.0±24.1, TNF-α(ng/L): 117.0±13.1 vs. 196.2±4.5], the expressions of p-TAK1 and p-NF-κBp65 were significantly decreased (p-TAK1/TAK1: 0.74±0.09 vs. 1.85±0.15, p-NF-κBp65/NF-κBp65: 1.15±0.17 vs. 2.36±0.20), with statistically significant differences (all P <0.05). There was no significant difference in the p-JNK, p-p38MAPK expressions between FⅩa+RIV+LPS group and FⅩa+LPS group (p-JNK/JNK: 1.64±0.12 vs. 1.65±0.15, p-p38MAPK/p38MAPK: 2.31±0.32 vs. 2.35±0.20, both P > 0.05). Conclusion Rivaroxaban can effectively relieve the inflammatory response of HUVEC stimulated by LPS, which may be related to the inhibition of NF-κB signaling pathway activation rather than MAPK signaling pathway.
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ObjectiveTo probe the indication,treatment algorithm,anesthesia method,safety and efficacy ofextracorporealshockwavelithotripsy(ESWL)incombinationwithendoscopicretrograde cholangiopancreatography (ERCP) for pancreatic duct stones.MethodsThe patients with chronic pancreatitis and large pancreatic duct stones ( ＞ 5 mm diameter) and receiving ESWL and ERCP between March and July 2011 in Changhai Hospital were prospective studied.The third generation of extracorporeal shockwave lithotripsy was applied in ESWL,and the localization of stone was determined by X-ray.No more than 5000 shocks were given per session,and ESWL was performed continuously till the calculi were fragmented,and then was cleared by ERCP.ResultsA total of 100 patients underwent ESWL during the 5 months,among whom 84 patients received ERCP treatment and 41 cases failed to deep cannulation (41/84,48.8％ ).Multiple stones were seen in 83 patients.Ninety five patients had radio-opaque stones,two patients had radiolucent calculi,while three patients had both radio-opaque and radiolucent stones.Seventy five percent,14％ and 11％ stones were located in pancreatic head,pancreatic head and body,pancreatic body and tail,respectively.A total of 175 ESWL procedures were performed,43 patients needed 2 or more sessions for successful fragmentation.Anesthesia method was mainly intravenous sedation,accounting for 96％ (168/175).ERCP was successful in 96 patients after ESWL,only 4 patients failed after ESWL. Forty one cases which failed ERCP procedures before ESWL underwent ERCP,and 37 patients (90.2％) achieved successful cannulation.Successful fragmentation ratewas 100％.Complete clearance was achieved in 78 patients,and complication rate of post-ERCP pancreatitis,fever was 1.71％ (n =3 ),0.57％ (n =1 ),and the overall complication rate was 2.28％.Conclusions ESWL is an effective,safe and necessary modality for fragmentation of large PD stones in the management of minimal invasive treatment of chronic pancreatitis.
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Objective To evaluate the safety and efficacy of therapeutic ERCP for patients above 90 years of age.Methods The data of 37 patients of above 90 years who underwent 42 ERCP procedures from January 2001 to December 2009 were studied retrospectively and compared with those of 152 matched patients ( 168 procedures) below 65 years old at a 1∶4 ratio for success rate and complications.Results The rate of complete success,partial success,and failure in observation group was 73.81％ (31/42),19.05％(8/42) and 2.38％ (1/42),respectively,which were similar (P ＞0.05) with those in control group,with complete success rate at 85.12％ ( 143/168),partial success rate at 12.50％ (21/168) and failure rate at 2.38％ (4/168).The rate of terminated operation in observation group (4.76％,2/42) was significantly higher than that of the control group (0.00％,0,P =0.039).The overall rate of complication in observation group was 7.14％ ( 3/42 ),slightly higher than that of the control group ( 6.55％,11/168,P ＞0.05 ).There was no significant difference between the two groups regarding the rates and severity of such complications as pancreatitis,hemorrhage and infection ( P ＞ 0.05 ).No perforation or death was observed.Conclusion Therapeutic ERCP for patients of 90 years or older is safe and effective.Adverse events related to chronic concomitant diseases need early detection and proper management.