Effects of aspirin on pancreatic cancer cells PANC-1 and its potential molecular mechanism.

PURPOSE: To explore the effects of aspirin on the proliferation and apoptosis of human pancreatic cancer cells and its potential molecular mechanism. METHODS: This study included patients with pancreatic cancer who were divided into experimental group and control group. The cell proliferation ability was detected via cell counting kit-8 (CCK-8) assay and colony forming ability via colony formation assay. In addition, changes in proteins in the phosphatidyl inositol-3-kinase (PI3K)/protein kinase B (Akt)/mammalian target of rapamycin (mTOR) pathway were assessed using Western blotting, and rescue experiment was conducted to investigate whether aspirin can affect cell proliferation by inhibiting the PI3K/Akt/mTOR signaling pathway. RESULTS: The results of CCK-8 assay showed that the proliferation rate of PANC-1 cells was decreased in a time- and dose-dependent manner after they were treated with aspirin at different concentrations. Colony formation assay confirmed that cell colony forming ability was significantly reduced with the increase in aspirin treatment concentration (p<0.05). Besides, the apoptosis rate and the number of cells in the experimental group were higher and larger than those in the control group (p<0.05). According to Western blotting results, the protein expressions of PI3K, phosphorylated (p)-Akt and p-mTOR were decreased after aspirin treatment. Rescue experimental results manifested that insulin-like growth factor 1 (IGF-1) supplementation remarkably elevated the expressions of PI3K, p-Akt and p-mTOR compared with phosphate-buffered saline (PBS) supplementation. It was found in CCK-8 assay that IGF-1 supplementation markedly reversed the inhibition of aspirin on the proliferation of PANC-1 cells in comparison with PBS supplementation. CONCLUSIONS: Aspirin inhibits the proliferation and promotes the apoptosis of pancreatic cancer cells by inactivating the PI3K/Akt/mTOR signaling pathway.

Protective effect of ulinastatin on hepatic ischemia reperfusion injury through autophagy activation in Chang liver cells.

This study aimed to investigate the protective effect of ulinastatin in hepatic ischemia-reperfusion progress, involving its association with the role of autophagy during hypoxia-induced hypoxia-reoxygenation injury in vitro. The model of hepatic hypoxia/reoxygenation (H/R) injury in Chang liver cells was established. After treatment with ulinastatin at the doses of 10, 100, and 1000 U/mL in H/R liver cells, the cell proliferation was significantly increased, morphological damage was reduced, and the cell apoptosis rate was decreased. The protein levels of antiapoptotic myeloid cell leukemia-1 (Mcl-1) and caspase-3 were upregulated, and C-PARP protein was downregulated. Meanwhile, ulinastatin led to an increase in the messenger RNA and protein levels of autophagy maker Unc-like kinase 1 (ULK1), Beclin-1, and microtubule-associated protein 1 light chain 3 (LC-3) and a decrease in p62. Then, 3-methyladenine (3-MA), an inhibitor of autophagy, made morphological damage and cell apoptosis worsen in ulinastatin-treated H/R liver cells. And the expression levels of caspase-3, C-PARP, p62, Beclin-1, and LC-3, proteins were also reversed by 3-MA. Taken together, our results demonstrate that ulinastatin inhibited the hepatic H/R injury in Chang liver cells, which was, to some extent, related to the autophagy activation.

The predictive value of serial changes in diaphragm function during the spontaneous breathing trial for weaning outcome: a study protocol.

INTRODUCTION: There is a variety of tools being used in clinical practice for the prediction of weaning success from mechanical ventilation. However, their diagnostic performances are less than satisfactory. The purpose of this study is to investigate the value of serial changes in diaphragm function measured by ultrasound during the spontaneous breathing trial (SBT) as a weaning predictor. METHODS AND ANALYSIS: This is a prospective observational study conducted in a 10-bed medical emergency intensive care unit (EICU) in a university-affiliated hospital. The study will be performed from November 2016 to December 2017. All patients in the EICU who are expected to have mechanical ventilation for more than 48 hours through endotracheal tube are potentially eligible for this study. Patients will be included if they fulfil the criteria for SBT. All enrolled patients will be ventilated with an Evita-4 by using volume assist control mode prior to SBT. Positive end-expiratory pressure (PEEP) will be set to 5 cmH2O and fractional inspired oxygen (FiO2) will be set to a value below 0.5 that guarantees oxygen saturation by pulse oximetry (SpO2) greater than 90%. Enrolled patients will undergo SBT for 2 hours in semirecumbent position. During the SBT, the patients will breathe through the ventilator circuit by using flow triggering (2 L/min) with automatic tube compensation of 100% and 5 cmH2O PEEP. The FiO2 will be set to the same value as used before SBT. If the patients fail to tolerate the SBT, the trial will be discontinued immediately and the ventilation mode will be switched to that used before the trial. Patients who pass the 2-hour SBT will be extubated. Right diaphragm excursion and bilateral diaphragm thickening fraction will be measured by ultrasonography during spontaneous breathing. Images will be obtained immediately prior to the SBT, and at 5, 30, 60, 90 and 120 min after the initiation of SBT. Rapid shallow breathing index will be simultaneously calculated at the bedside by a respiratory nurse. ETHICS AND DISSEMINATION: The study protocol is approved by the ethics committee of Sir Run Run Shaw Hospital, an affiliate of Zhejiang University, Medical College. The results will be published in a peer-reviewed journal and shared with the worldwide medical community. TRIAL REGISTRATION NUMBER: ISRCTN42917473; Pre-results.

Bone marrow-derived mesenchymal stem cells (BMSCs) repair acute necrotized pancreatitis by secreting microRNA-9 to target the NF-κB1/p50 gene in rats.

Acute pancreatitis (AP) is a common acute abdominal disease, 10-20% of which can evolve into severe AP (SAP) causing significant morbidity and mortality. Bone marrow-derived mesenchymal stem cells (BMSCs) have the potential of repairing SAP, but the detailed mechanism remains unknown. We demonstrate here that microRNA-9 (miR-9) modified BMSCs (pri-miR-9-BMSCs) can significantly reduce the pancreatic edema, infiltration, hemorrhage, necrosis, the release of amylase and lipase. Meanwhile, decreased local/systemic inflammatory response (TNF-α↓, IL-1ß↓, IL-6↓, HMGB1↓, MPO↓, CD68↓, IL-4↑, IL-10↑, and TGF-ß↑) and enhanced regeneration of damaged pancreas (Reg4↑, PTF1↑, and PDX1↑) are also promoted. But these effects diminish or disappear after antagonizing miR-9 (TuD). Besides, we find that miR-9 is negatively correlated with AP and miR-9 agomir which can mimic the effects of pri-miR-9-BMSCs and protect injured pancreas. Furthermore, we investigate that BMSCs deliver miR-9 to the injured pancreas or peripheral blood mononuclear cell (PBMC), which can target the NF-κB1/p50 gene and inhibit the NF-κB signaling pathway (p-P65↓, NF-κB1/p50↓, IκBα↑, IκBß↑). Taken together, these results show that miR-9 is a key paracrine factor of BMSCs attenuating SAP targeting the NF-κB1/p50 gene and suppressing the NF-κB signaling pathway.

Intravenous hMSCs Ameliorate Acute Pancreatitis in Mice via Secretion of Tumor Necrosis Factor-α Stimulated Gene/Protein 6.

The administration of mesenchymal stem cells/multipotent mesenchymal stromal cells (MSCs) to enhance tissue repair is currently undergoing clinical trials. Some studies, including our previous work, have also revealed the beneficial effect of MSCs in severe acute pancreatitis (SAP); however, their mechanisms or mode of action remain controversial. In this study, we demonstrated that intravenously (i.v.)-administered human MSCs (hMSCs) remarkably promoted recovery from experimental SAP without significant engraftment of hMSCs in the damaged pancreas. Interestingly, we found that i.v.-administered hMSCs with knockdown of TSG-6 expression lost most of their anti-inflammatory effects and thus could not significantly ameliorate SAP. As expected, the effects of hMSCs were also duplicated by i.v. infusion of recombinant TSG-6. Furthermore, our results showed that the increase of oxidative stress, activation of the NLRP3 inflammasome and NF-κB signaling in SAP was substantially inhibited following administration of hMSCs or TSG-6, which was dependent on the presence of CD-44 receptors in acinar cells. In conclusion, our study, for the first time, revealed that novel mechanisms are responsible for the immunomodulatory effect of i.v. hMSCs.

Comparison of bupivacaine and parecoxib for postoperative pain relief after laparoscopic cholecystectomy: a randomized controlled trial.

BACKGROUND: Pain is the most common complaint of patients on the first day after laparoscopic cholecystectomy (LC). The aim of this study was to compare the efficacy of local anesthesia with bupivacaine and intravenous parecoxib on postoperative abdominal pain relief up to 24 h after surgery. METHODS: One hundred and eighty patients who underwent LC were randomized to one of three groups with sixty patients each: Group A received 50 mg 0.5% bupivacaine subcutaneously at trocar sites before incision closure; Group B received intravenous parecoxib (40 mg) after entering the recovery room; Group C did not receive postoperative analgesia unless needed and was served as control. The postoperative pain at 1, 2, 4, 8, 12, and 24 h after the operation was assessed using a visual analog scale (VAS). Secondary outcomes, including intraoperative and postoperative complications, the incidence of shoulder pain, pethidine requirements, postoperative nausea and vomiting, and hospital stay were also recorded. RESULTS: At 1, 2, and 4 hours after surgery, VAS pain scores were significantly lower in group A and B compared with group C (P < 0.05 for all). There was no significant difference among the three groups at 8, 12, and 24 hours after the procedure (P > 0.05 for all). A repeated-measures ANOVA analysis revealed that VAS pain scores over the first 24 hours after LC were significantly lower in group A and B compared with group C (P = 0.014 and P = 0.029 for between-group comparison, respectively). Furthermore, the percentage of patients requiring postoperative rescue analgesics was significantly higher in group C as compared with group A and group B (P = 0.018). CONCLUSION: Local anesthesia with bupivacaine and intravenous parecoxib are both effective at decreasing postoperative pain and pethidine requirements after LC.

Bone Marrow-Derived Mesenchymal Stem Cells Repair Necrotic Pancreatic Tissue and Promote Angiogenesis by Secreting Cellular Growth Factors Involved in the SDF-1 α /CXCR4 Axis in Rats.

Acute pancreatitis (AP), a common acute abdominal disease, 10%-20% of which can evolve into severe acute pancreatitis (SAP), is of significant morbidity and mortality. Bone marrow-derived mesenchymal stem cells (BMSCs) have been reported to have a potential therapeutic role on SAP, but the specific mechanism is unclear. Therefore, we conducted this experiment to shed light on the probable mechanism. We validated that SDF-1α significantly stimulated the expressions of VEGF, ANG-1, HGF, TGF-ß, and CXCR4 in BMSCs, which were inhibited by its receptor agonist, AMD3100. The capacities of proliferation, migration, and repair of human umbilical vein endothelial cells were enhanced by BMSCs supernatant. Meanwhile, BMSCs supernatant could also promote angiogenesis, especially after the stimulation with SDF-1α. In vivo, the migration of BMSCs was regulated by SDF-1α/CXCR4 axis. Moreover, transplanted BMSCs could significantly alleviate SAP, reduce the systematic inflammation (TNF-α↓, IL-1ß↓, IL-6↓, IL-4↑, IL-10↑, and TGF-ß↑), and promote tissue repair and angiogenesis (VEGF↑, ANG-1↑, HGF↑, TGF-ß↑, and CD31↑), compared with the SAP and anti-CXCR4 groups. Taken together, the results showed that BMSCs ameliorated SAP and the SDF-1α/CXCR4 axis was involved in the repair and regeneration process.

Primary closure and rate of bile leak following laparoscopic common bile duct exploration via choledochotomy.

BACKGROUND: Choledocholithiasis is traditionally managed by endoscopic retrograde cholangiopancreatography or T-tube insertion following common bile duct exploration. This study examined the efficacy and safety of primary duct closure following laparoscopic common bile duct exploration (LCBDE) via choledochotomy. METHODS: Between September 2011 and September 2013, 157 consecutive patients underwent LCBDE via choledochotomy. RESULTS: Of 157 LCBDE procedures, 138 (87.9%) were successfully completed with primary closure of the choledochotomy. Eight patients (5.1%) underwent closure with T-tube drainage after choledochotomy and 11 patients (7.0%) were converted to open surgery. The biliary tree was free of stones at the end of surgery in 154 patients (98.1%). Postoperative bile leak occurred in 6 patients (3.8%). The median follow-up period was 18 (2-33) months, with no evidence of further bile duct stones or bile duct stricture in any patients. Univariable analysis revealed that successful duct clearance (p = 0.010) and diameter of the common bile duct (p < 0.001) were two significant risk factors for bile leak. CONCLUSIONS: Primary duct closure following LCBDE is effective and safe for the management of choledocholithiasis. The postoperative bile leak rate may be low in skilled laparoscopic surgeons with a careful selection of patients.

Angiopoietin-1 gene-modified human mesenchymal stem cells promote angiogenesis and reduce acute pancreatitis in rats.

Mesenchymal stem cells (MSCs) can serve as a vehicle for gene therapy. Angiopoietin-1 (ANGPT1) plays an important role in the regulation of endothelial cell survival, vascular stabilization, and angiogenesis. We hypothesized that ANGPT1 gene-modified MSCs might be a potential therapeutic approach for severe acute pancreatitis (SAP) in rats. Human umbilical cord-derived MSCs with or without transfection with lentiviral vectors containing the ANGPT1 gene were delivered through the tail vein of rats 12 h after induction of SAP. Administration of MSCs alone significantly reduced pancreatic injury and inflammation, as reflected by reductions in pancreatitis severity scores and serum amylase and lipase levels as well as reducing the serum levels of proinflammatory cytokines (TNF-α, IFN-γ, IL-1ß, and IL-6). Furthermore, administration of ANGPT1-transfected MSCs resulted in not only further reductions in pancreatic injury and serum levels of proinflammatory cytokines, but also promotion of pancreatic angiogenesis. These results suggest that MSCs and ANGPT1 have a synergistic role in the treatment of SAP. ANGPT1 gene-modified MSCs may be developed as a potential novel therapy strategy for the treatment of SAP.

Hand-assisted versus conventional laparoscopic splenectomy: a systematic review and meta-analysis.

BACKGROUND: Hand-assisted laparoscopic splenectomy (HALS) has been widely applied for the resection of larger spleens. We conducted a systematic review and meta-analysis to evaluate the safety and feasibility of HALS compared with conventional laparoscopic splenectomy (CLS). METHODS: A comprehensive literature search in MEDLINE, EMBASE and Cochrane Library databases was performed to compare clinical outcomes of CLS and HALS. Data were extracted by two independent reviewers. Pooled odds ratios and weighted mean differences with 95% confidence intervals were calculated by meta-analytic software. RESULTS: Nine non-randomized controlled studies for a total of 463 patients were selected to satisfy the inclusion criteria (HALS versus CLS: 170 versus 293, respectively). The groups were similar in operative time, estimated operative blood loss, length of hospital stay, mortality and intraoperative and post-operative complications. There was a significantly reduced conversion rate in the HALS versus CLS group (odds ratio: 2.98; 95% confidence interval 1.28 to 6.93; P = 0.01). Splenic weights in the HALS group were higher than in the CLS group (weighted mean differences: -0.93; 95% confidence interval -1.74 to -0.11; P = 0.03). CONCLUSION: HALS may be preferable to CLS for the treatment of patients with enlarged spleens. The result needs to be certified by further random controlled trials.

Clinical comparison of distal pancreatectomy with or without splenectomy: a meta-analysis.

OBJECTIVE: A distal pancreatectomy has routinely been used for removing benign/borderline malignant tumors of the body and tail of the pancreas; however, controversy exists whether or not the spleen should be saved. Therefore, we conducted this meta-analysis for comparing the clinical outcomes of patients who underwent distal pancreatectomy with or without splenectomy. METHODS: A literature research from the databases of Medline, Embase, and Cochrane library was performed to evaluate and compare the clinical outcomes between spleen-preserving distal pancreatectomy (SPDP) and distal pancreatectomy with splenectomy (DPS). Pooled odds ratio (OR) and weighted mean difference (WMD) with 95% confidence interval (95% CI) were calculated using fixed-effects or random-effects models. RESULTS: Eleven non-randomized controlled studies involving 897 patients were selected to satisfy the inclusion criteria; 355 patients underwent SPDP and 542 patients underwent DPS. Compared with DPS, SPDP required a shorter hospital stay (WMD = 1.16, 95% CI = -2.00 to -0.31, P = 0.007), and had a lower incidence of intra-abdominal abscesses (OR = 0.48, 95% CI = 0.27 to 0.83, P = 0.009). In addition, spleen infarctions occurred in SPDP, most of which involved use of the Warshaw method for preserving the spleen. There were no differences between the SPDP and DPS groups with respect to operative time, operative blood loss, requirement for blood transfusion, pancreatic fistulas, thromboses, post-operative bleeding, wound infections and re-operation rates. CONCLUSION: SPDP should be performed due to the benefits of the immune system and quick post-operative recovery. It is also essential to preserve the splenic artery and vein. Large randomized controlled trials are further needed to verify the results of this meta-analysis.