Objective:To explore the regulatory effect of cellular FLICE-like inhibitory protein (cFLIP) on myocardial ischemia-reperfusion injury based on the RIPK1/RIPK3/MLKL-mediated necroptosis pathway.Methods:The cardiomyocyte hypoxia/reoxygenation (H/R) model was constructed by hypoxia for 4 h/reoxygenation for 12 h, and the rat ischemia reperfusion (I/R) model was constructed by ligating the left anterior descending artery for 30 min and reperfusion for 3 h. CCK-8 method was used to detect the viability of cardiomyocytes in each group. DAPI/PI double staining was used to observe changes in necrosis rate of myocardial cell. STRING database was used to predict the protein interaction network of cFLIP. TTC staining was used to detect the area of myocardial infarction in each group of rats, and the protein expression of cFLIPL, cFLIPS, p-RIPK1, p-RIPK3 and p-MLKL were detected by Western blot.Results:In cardiomyocyte H/R injury and myocardial tissue I/R injury, the protein expressions of cFLIPL and cFLIPS were significantly down-regulated, while the levels of p-RIPK1, p-RIPK3 and p-MLKL were increased significantly. Up-regulating the protein expression of cFLIPL and cFLIPS could significantly reduce the damage of cardiomyocytes and the rate of cell necrosis induced by H/R, and decrease the area of myocardial infarction caused by I/R. STRING database results showed that cFLIP had direct protein interactions with RIPK1 and RIPK3. Overexpression of cFLIP in cardiomyocyte and myocardial tissue significantly inhibited H/R or I/R induced the phosphorylation levels of RIPK1, RIPK3 and MLKL.Conclusions:Overexpression of cFLIP can significantly inhibit the RIPK1/RIPK3/MLKL-mediated necroptosis, thereby reducing myocardial cell damage and decreasing the area of myocardial infarction.

Objective:To explore the influences of different lying positions on vital signs and comforts in patients with tracheal intubation removed after intestinal fistula surgery.Methods:From October 2018 to June 2019, convenience sampling was used to select 150 patients with intestinal fistula surgery after general anesthesia in Intensive Care Unit of the General Hospital of Eastern Theater Command were selected as the research object. After waiting for the patient to wake up from anesthesia, the tracheal intubation was removed, and the lying position was changed after normal spontaneous breathing through the nose. Patients were randomly divided into 5 groups, 30 cases in each group. Group A was in the supine position; group B was in the lying position with the head of the bed raised by 15°; group C was in the lying position with the head of the bed raised by 30°; group D was in the lying position with the head of the bed raised by 45°; group E was in the lying position with the head of the bed raised by 15° to 45 °. At the beginning, the head of the bed was shaken to 15 °; and after two hours, the head of the bed was shaken to 30 °, and after another two hours, the head of the bed was shaken to 45 °. After 5 groups of patients fixed the angle of the lying position, when the patient complained of discomfort or the body position changed spontaneously, the patient changed the lying position. The duration of the lying position, uncomfortable symptoms and pain degree of abdominal incision were compared among 5 groups.Results:Patients with tracheal intubation removed after intestinal fistula surgery had a longer persistence in the 15° and 30° lying positions within 6 hours, respectively (72.27±12.41) min and (69.37±9.10) min. There were statistically significant differences in the persistence time of 5 different lying positions ( P<0.01) . The number of patients in group B and C with waist pain was less, and there were more patients with painless incisions, and there were statistically significant differences among 5 groups ( P<0.05) . Conclusions:Among patients with tracheal intubation removed and normal spontaneous breathing through the nose after intestinal fistula surgery, the 15° to 30° lying position within 6 hours after the operation can prolong the patient's adherence to the lying position, reduce the incidence of abdominal incision pain and waist pain, and reduce the probability of dizziness, nausea and vomiting.

Objective·To explore the biologic effect and mechanism of adenanthin (Aden) on multiple myeloma (MM) cells. Methods·MM cells, H929 and U266 were treated with various dose of Aden for different time, and the density and viability of MM cells were detected by trypan blue exclusion assay. After H929 and U266 cells were treated with various dose of Aden for 24 hours, cell growth inhibition was examined by CCK8 assay, and cell apoptosis was examined by AnnexinV-APC/PI staining assay. Apoptosis related proteins, NF-κB signaling pathway associated proteins and the NF-κB regulated proteins were detected by Western blotting. The effect of Aden on the thermal stability of IKKβ protein was determined by CETSA assay. Results·Trypan blue exclusion results showed that Aden inhibited cell growth and reduced cell viability in concentration and time dependent manners. U266 was more sensitive than H929 when exposed to the same concentration of Aden. The CCK8 results showed that Aden inhibited the growth of H929 and U266 cells in a concentration dependent manner. Flow cytometry results suggested that Aden induced a low apoptosis rate of MM cells. Moreover, cleavage of caspase3 and PARP were detected in U266 cells but not in H929 cells. CETSA assay indicated that Aden decreased the thermal stability of IKKβ. Expression of p-p65 and p-IκBα proteins decreased in MM cells treated with Aden. Conclusion·Aden significantly inhibits MM cell proliferation by inhibiting NF-κB activation through interacting with IKKβ. Aden has little effect on apoptosis of MM cells.

Objective·To establish a cell-based screening system for identification of compounds with activity in regulating retinoic acid receptor (RARα) stability. Methods·The modified pMSCV plasmid constructs, named as RARα-EGFP-IRES-DsRed, consists of enhanced green fluorescent protein (EGFP) fusing to RARα and red fluorescent protein (DsRed) as internal references incorporating the internal ribosome entry site (IRES) as interval sequence. The RARα-EGFP-IRES-DsRed plasmid was stably transfected into NB4 cells which were named as NB4-pMGIR-RARα. Fluorescence signals of EGFP and DsRed indirectly reflecting the expression of RARα, were detected by flow cytometry in cells that were treated with all-trans retinoic acid, sodium valproate, cytarabine, lenalidomide, etoposide, montelukast and gambogic acid, respectively. Effects of these compounds on the expression of RARα protein were further examined by Western blotting. Results·A double fluorescence reporter system for screening compounds that can increase the stability of RARα protein was successfully established, and sodium valproate was identified as a potent compound to promote the stability of RARα. Conclusion·The double fluorescence reporter system can be used to screen compounds regulating the stability of RARα protein, which can be further used to identify compounds regulating the stability of other proteins.

OBJECTIVETo explore the efficacy of over-the-scope clip (OTSC) in the treatment of gastrointestinal fistula.

METHODSClinical data of 12 gastrointestinal fistula patients, including 3 internal fistula and 9 external fistula treated with OTSC in our institute from March 2015 to May 2016 were retrospectively analyzed. OTSC was performed when pus was drained thoroughly and intra-abdominal infection around gastrointestinal fistula was controlled, and each patient received one clip to close fistula.

RESULTSThere were 6 female and 6 male patients with mean age of (50.1±12.6) years. The successful rate of endoscopic closure was 100% without complications including bleeding and intestinal obstruction during and after OTSC treatment. According to comprehensive evaluation, including drainage without digestive juices, no recurrence of intra-abdominal infection, no overflow of contrast medium during digestive tract radiography, and CT examination without intra-abdominal abscess, clinical gastrointestinal fistula closure was 91.7%(11/12). There was no recurrence of gastrointestinal fistula during 3 months of follow-up in 11 patients. In the remaining 1 case, the gastric fistula after laparoscopic sleeve gastrectomy recurred one week after OTSC treatment because of intra-abdominal infection surrounding fistula, and was cured by surgery finally.

CONCLUSIONThe endoscopic closure treatment of OTSC for gastrointestinal fistula is successful and effective, and control of intra-abdominal infection around fistula with adequate drainage is the key point.

Adult ; Digestive System Fistula ; complications ; drug therapy ; surgery ; Drainage ; Endoscopy, Gastrointestinal ; instrumentation ; methods ; Female ; Humans ; Intraabdominal Infections ; etiology ; therapy ; Male ; Middle Aged ; Recurrence ; Retrospective Studies ; Suppuration ; therapy ; Surgical Fixation Devices

Objective:To explore therapeutic effects of sodium nitroprusside (SNP) combined verapamil on no-reflow during percutaneous coronary intervention (PCI).Methods: A total of 106 patients, who suffered from no-reflow during PCI in our department from Jan 2011 to Dec 2013, were selected.According to random number table method, patients were divided into SNP group (n=55, received SNP based on routine treatment) and combined treatment group (n=51, received verapamil based on SNP group).Cardiac troponin I (cTnI) level before and 16h~18h after PCI, cardiac function indexes after 12-month follow-up, incidence of major adverse cardiovascular events (MACE) were measured and compared between two groups.Results: Compared with before PCI, there were significant rise in cTnI level in both groups on 16~18h after PCI, P=0.001 both;compared with SNP group, there were significant reductions in cTnI level [(1.31±0.44)μg/L vs.(0.11±0.02)μg/L] and percentage of cTnI>0.10μg/L (94.5% vs.54.9%) in combined treatment group, P=0.001 both.Compared with SNP group after 12 months, there was significant rise in left ventricular ejection fraction [(62.29±3.06)% vs.(65.65±3.94)%], and significant reductions in left ventricular end-diastolic dimension[(50.24±3.73)mm vs.(47.60±4.72)mm] and left ventricular end-systolic dimension [(33.29±2.11)mm vs.(31.00±4.33)mm] in combined treatment group, P<0.05 all.There were no significant adverse reactions during hospitalization and follow-up in both groups.Conclusion: When no-reflow occurs during PCI, intracoronary injection of SNP combined verapamil can improve cardiac function, and its safety is good, which is worth extending.

Aim To observe the effect of mineralocor-ticoid receptor blockade eplerenone on cell proliferation in obstructed kidney of rats. Methods Renal intersti-tial fibrotic animals were made with unilateral ureteral obstruction (UUO) and treated with eplerenone100 mg · kg - 1 · d - 1 . The kidneys were harvested on the 10th day and proliferating cell nuclear antigen ( PC-NA ), serum and glucocorticoid induced kinase-1 (SGK-1 ) and transforming growth factor-β1 ( TGF-β1 ) were detected with immunohistochemistry and Western blot. Results Renal histopathology showed large quantities extracellular matrix (ECM) accumula-tion in kidney with UUO, large numbers of inflammato-ry cells infiltrated in renal interstitium, renal tubular expansion and exfoliation of epithelial cells . The cell proliferation and ECM accumulation were inhibited in eplerenone treated rats significantly. Immunohisto-chemistry and Western blot showed that expressions of PCNA,SGK-1 and TGF-β1 were significantly up-regu-lated with UUO and down-regulated by eplerenone. Conclusion Eplerenone plays the role in inhibiting the cell proliferation and reducing ECM accumulation by down-regulating expression of SGK-1 pathway in rats with unilateral ureteral obstruction.

OBJECTIVETo investigate the risk factors of surgical site infection (SSI) in definitive surgery of intestinal fistulas.

METHODSPatients with gastrointestinal fistula undergoing definitive operation during November 2011 to November 2013 in Jinling Hospital were prospectively enrolled in the study. Risk factors of SSI were analyzed. Patients' characteristics, surgery-related data and fistula-related data were prospectively collected. Risk factors of SSI were analyzed.

RESULTSA total of 191 cases were enrolled and 51 cases developed SSI. Univariate analysis showed that patients with risk index category (RIC)≥2, length of abdominal incisions>15 cm, and duration of drainage tubes>10 days had significantly higher incidence of SSI (P<0.05). Multivariate Logistics analysis demonstrated that RIC and duration of drainage tube were independent risk factors for SSI (P=0.02, P=0.01, respectively).

CONCLUSIONSRIC≥2 and duration of drainage tubes>10 days are independent risk factors for development of SSI.

Humans ; Incidence ; Intestinal Fistula ; Multivariate Analysis ; Risk Factors ; Surgical Wound Infection

Objective To compare the efficacy and safety of singular double antithrombotie therapy (DT) using warfarin plus clopidogrel and the combined antithrombotie therapy of 3-month triple antithrombotie therapy (TT) using warfarin, aspirinand clopidogrel and 9-month double antithrombotie therapy (DT) for the patients with atrial fibrillation undergoing PCI. Methods Ninety patients with atrial fibrillation undergoing PCI were randomly divided into two groups evenly: one group was treated with dual antithrombotic therapy group (DT) and the other group with the combined therapy, e. g. 3-month triple antithrombotie therapy (TT) and 9-month double antithrombotie therapy (DT + TT for short). All patients were followed-up by 12 months. The two groups were compared in terms of incidences of death , myocardial infarction , stroke , target-vessel revascularisation , stent thrombosis and bleeding adverse events. Results The incidences of myocardial infarction, stroke, target-vessel revascularisation , stent thrombosis and bleeding adverse events in the TT + DT group were all significantly lower than the DT group (P < 0.05). The follow-up on the safety indicated that the rate of bleeding in the TT +DT group was insignificantly higher than the DT group (P > 0.05). Conclusion There is no significant difference in safety between the two groups. However, the therapy of TT + DT is more effective.

Objective To investigate the risk factors of surgical site infection (SSI) in definitive surgery of intestinal fistulas. Methods Patients with gastrointestinal fistula undergoing definitive operation during November 2011 to November 2013 in Jinling Hospital were prospectively enrolled in the study. Risk factors of SSI were analyzed. Patients'characteristics, surgery-related data and fistula-related data were prospectively collected. Risk factors of SSI were analyzed. Results A total of 191 cases were enrolled and 51 cases developed SSI. Univariate analysis showed that patients with risk index category (RIC)≥2, length of abdominal incisions > 15 cm, and duration of drainage tubes > 10 days had significantly higher incidence of SSI(P<0.05). Multivariate Logistics analysis demonstrated that RIC and duration of drainage tube were independent risk factors for SSI (P=0.02, P=0.01, respectively). Conclusions RIC≥2 and duration of drainage tubes > 10 days are independent risk factors for development of SSI.