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Objective:To investigate the function of gasdermin D (GSDMD) in intestinal damage of mice with severe acute pancreatitis (SAP).Methods:The healthy C57BL/6 mice were divided into four groups randomly, including normal saline (NS) group, small interfering RNA (siRNA)-NS group, SAP model group and siRNA-SAP group, with 6 mice in each group. The SAP mouse model was reproduced by intraperitoneal injection of caerulein 50 μg/kg combined with lipopolysaccharide (LPS) 10 mg/kg; the NS group was given the same amount of NS; in the siRNA-SAP group and siRNA-NS group, siRNA 50 mg/kg was injected through the tail vein three times before modeling or injection of NS. The blood of mice eyeball in each group was taken 12 hours after modeling, and serum interleukins (IL-1β, IL-18) levels were detected by enzyme linked immunosorbent assay (ELISA). The mice were sacrificed to observe the general changes in abdominal cavity, the pancreas and ileum tissues were taken to observe the pathological changes under a light microscope. The expression of long-chain non-coding RNA uc.173 (lnc uc.173) was detected by reverse transcription-polymerase chain reaction (RT-PCR). Immunohistochemical method was used to detect the expression of tight junction proteins zonula occluden-1 (ZO-1) and Occludin in intestinal mucosal epithelial cells. Western blotting was used to detect the GSDMD protein expression level in the intestinal tissue.Results:The serum levels of IL-1β and IL-18 in the SAP model group were significantly higher than those in the NS group and the siRNA-NS group [IL-1β (ng/L): 146.66±1.40 vs. 44.48±5.76, 81.49±10.75, IL-18 (ng/L): 950.47±177.09 vs. 115.43±16.40, 84.84±21.90, all P < 0.05]; and the levels of IL-1β and IL-18 in the siRNA-SAP group were significantly lower than those in the SAP model group [IL-1β (ng/L): 116.26±15.54 vs. 146.66±1.40, IL-18 (ng/L): 689.96±126.08 vs. 950.47±177.09, both P < 0.05]. General observation showed that there were no obvious abnormalities in the abdominal cavity of the mice in the NS and siRNA-NS groups; the mice in the SAP model group and the siRNA-SAP group had different degrees of edema and congestion in the intestine; compared with the SAP model group, the abnormalities in the siRNA-SAP group was significantly reduced. Under light microscope, there were no obvious changes in the pancreas and intestinal mucosa in the NS group and the siRNA-NS group; the pancreatic tissue of the SAP model group and the siRNA-SAP group had different degrees of edema, inflammatory cell infiltration, and lobular structure damage, and the intestinal mucosa was damaged to a certain degree, and the villi were broken to varying degrees, but the damage in the siRNA-SAP group was lighter. The results of RT-PCR showed that the expression of lnc uc.173 in the intestinal tissues of the model SAP group was significantly lower than that of the NS group and the siRNA-NS group (2 -ΔΔCt: 0.26±0.12 vs. 1.01±0.37, 0.67±0.32, both P < 0.05), while the expression of lnc uc.173 in the siRNA-SAP group was significantly higher than that in the SAP model group (2 -ΔΔCt: 0.60±0.39 vs. 0.26±0.12, P < 0.05). Immunohistochemistry showed that ZO-1 and Occludin proteins in the NS group were distributed along the epithelial cells of the intestinal mucosa, showing a strong expression; ZO-1 and Occludin expressions were significantly reduced in the SAP model group and siRNA-SAP group, but the expressions in the siRNA-SAP group was higher than that in the SAP model group. Western blotting showed that the expression level of GSDMD protein in the intestinal tissues of the SAP model group was significantly higher than that of the NS group and the siRNA-NS group [GSDMD protein (GSDMD-N/β-actin): 1.99±0.46 vs. 1, 1.00±0.78, both P < 0.05]. Compared with the SAP model group, the expression of GSDMD protein in the siRNA-SAP group was significantly decreased [GSDMD protein (GSDMD-N/β-actin): 1.42±0.42 vs. 1.99±0.46, P < 0.05]. Conclusions:The systemic inflammatory response and intestinal mucosal barrier damage of SAP mice may be related to the increase of GSDMD expression in intestinal tissues. GSDMD mediates cell pyrolysis to promote the release of inflammatory factors, cause intestinal injury, and down-regulate the expression of intestinal epithelial cell tight junction proteins such as ZO-1 and Occludin, resulting in intestinal mucosal damage.
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Objective:To investigate the effect of ultrasound-guided midline catheter placement on the incidence of catheter-related bloodstream infection (CRBSI) in severe emergency patients.Methods:Five hundred and twenty-nine patients were chosen as the research objects from March 2018 to December 2019 at Emergency Intensive Care Unit, which was divided into the experimental group ( n=278) and the control group ( n=251). In the experimental group, ultrasound-guided midline catheter was used as central venous catheter (CVC) removal method of sequential, and in the control group, peripheral venous indwelling needle was used as sequential method after removal of CVC. CVC, midline catheter and the indwelling time of indwelling needle were counted. The utilization rate of CVC was compared between the two groups. Kaplan-Meier survival curve was plotted to describe the CVC indwelling time of the two groups and log-rank test was performed. Cox regression analysis was performed to analyze the influencing factors of CVC indwelling time and compare the incidence of CRBSI and other catheter-related complications. Results:The CVC indwelling time of the experimental group was significantly shorter than that of the control group (8 d vs. 13 d, P=0.000). The CVC utilization rate of the experimental group was significantly lower than that of the control group (49.83% vs. 80.45%, P=0.000). Multivariate Cox regression analysis showed that difficult intravenous access, length of ICU stay, the site of catheter placement, and midline catheter implantation without ultrasound-guidance were independent risk factors for prolonged CVC indwelling time ( P=0.000). The CRBSI rate of the experimental group was significantly lower than that of the control group (0.571‰ vs. 3.802‰, P=0.038). There was no significant difference in the incidence of other catheter-related complications between the two groups ( P=0.403). Conclusions:Ultrasound-guided midline catheter implantation can shorten the indwelling time of CVC, reduce the utilization rate of CVC, and reduce the incidence of CRBSI, which is worthy of clinical promotion.
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Objective:To analyze the risk factors of renal function recovery in patients with severe acute pancreatitis (SAP) combined with acute kidney injury (AKI).Methods:A retrospective study was conducted in 105 SAP patients with AKI who were admitted to ICU or EICU of the Affiliated Hospital of Qingdao University from January 2013 to October 2019. According to the recovery of renal function at 28 days, the patients were divided into the renal function recovery group and the poor recovery group. Multivariate logistic regression analysis was used to analyze the clinical data of the two groups and to determine the risk factors related to renal function recovery.Results:According to the recovery of renal function, 105 patients were divided into the renal function recovery group ( n=73) and the poor recovery group ( n=32). Compared with the renal function recovery group, patients in the poor recovery group were older, had a higher prevalence of diabetes and coronary heart disease and a higher score on the acute physiology and chronic health evaluation Ⅱ(APACHE Ⅱ); More patients had abdominal necrosis infection and abdominal hemorrhage. The proportion of patients who applied mechanical ventilation was higher in the poor recovery group. Multivariate logistic regression analysis showed that abdominal necrosis infection ( OR=5.088, 95% CI:1.041-24.871, P=0.044) and mechanical ventilation ( OR=4.615, 95% CI:1.126-18.904, P=0.034) were the independent risk factors of renal function recovery in SAP patients with AKI. Conclusions:Abdominal necrosis infection and mechanical ventilation are the independent risk factors for renal function recovery in patients with SAP and AKI.
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Objective:To investigate the effects of poly (ADP-ribose) polymerase-1(PARP-1) in intestinal mucosal barrier injury in rat model with severe acute pancreatitis (SAP).Methods:Twenty healthy male Wistar rats were divided into four groups ( n=5 each group) using a random table method: control, SAP, 3-aminobenzamide (3-AB), and 3-AB control groups. The SAP model was induced by intraperitoneal injection of cerulean with lipopolysaccharide. At 30 min, the rats were treated with the PARP-1 inhibitor, 3-AB, or normal saline,separately. After 12 h, all rats were sacrificed to harvest pancreas tissues, intestines tissues, and blood from the hearts for index detection. Serum amylase (AMY) and interleukin (IL)-6 levels were measured using an automatic biochemical instrument and enzyme-linked immunosorbent assay (ELISA), respectively.The protein expression of PARP-1 and nuclear factor (NF-κB) were measured using Western blot and that of occludin was measured using an immunohistochemical test. One-way analysis of variance was used for comparison of multiple groups of variables. Non-parametric tests of rank conversion were used when variances were not uniform. A P <0.05 was considered statistically significant. Results:Compared to the control group, the following indexes in the SAP group were significantly increased: ascites (with serious hemorrhage and necrosis in the pancreas and disordered intestinal villi),serum AMY and IL-6 levels, and the expression of PARP-1 and NF-κB. However, Occludin expression was significantly decreased. There was no significant difference between 3-AB group and 3-AB control group. Compared to the SAP group, the severity of SAP and pancreatitis-associated intestinal injury was significantly attenuated with the administration of 3-AB. Serum AMY and IL-6 levels were significantly decreased (serum AMY: 1 879.25 ± 736.6 U/L vs 5 569.33 ± 1993.48 U/L; IL-6: 77.98 ± 20.65 pg/mL vs 209.14 ± 79.08 pg/mL, both P<0.05), but the expression of PARP-1 and NF-κB were significantly increased (PARP-1: 1.44 ± 0.09 vs 1.49 ± 0.13; NF-κB: 0.63 ± 0.09 vs 0.96±0.08, both P<0.05). Similarly, Occludin expression was significantly decreased (6.7±1.5 vs 3.2±1.1, P<0.05). Conclusions:Inhibition of PARP-1 has protective effects on SAP associated intestinal mucosal barrier damage. The mechanism may be related to the inhibition of NF-κB signaling pathway and increase intestinal mucosal Occludin protein expression.
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Objective To find the pathogenies and risk factors related to surgical intensive care unit (SICU) readmission for patients who underwent hepatobiliary-pancreatic surgery, and to develop a predictive model for determining patients who are likely to be readmitted to SICU. Methods The patients who admitted to SICU of the Affiliated Hospital of Qingdao University from January 2013 to August 2018; who first stayed in SICU after hepatobiliary-pancreatic surgery; who were assessed and discharged from SICU by surgeons and SICU physicians after treatment, and then transferred to SICU again because of the change of their condition were enrolled. The unintended return to SICU within 3 days and 7 days were recorded. Patients who returned to SICU within 7 days were studied for the pathogenies, risk factors and predictive model of returning to SICU, and non-returning patients were enrolled according to 1∶1 as the controls. A total of 43 indicators were divided into five categories, including general clinical data, medical history, surgical indicators before first admission of SICU, length of first SICU stay, and other indicators on the day of first discharge from the SICU. Logistic regression was used to screen the risk factors associated with SICU readmission, then the Nomogram diagram was drawn by using the R 3.4.1 software for predicting SICU readmission, and the classification performance of Nomogram was evaluated by self-help sampling test. Results Of the 763 patients discharged from the SICU, 2.10% (16/763) of them were readmitted within 3 days and 3.28% (25/763) were readmitted within 7 days to the SICU unexpectedly. The pathogenies of SICU readmission within 7 days included infection [56.00% (14/25)], heart failure [16.00% (4/25)], infarction [12.00% (3/25)], bleeding [12.00% (3/25)], and sutures splitting [4.00% (1/25)]. The pathogenies of SICU readmission within 3 days included infection [56.25% (9/16)], heart failure [18.75% (3/16)], infarction [12.50% (2/16)], and bleeding [12.50% (2/16)]. Nomogram analysis showed that the risk factors associated with unplanned SICU readmission were length of first SICU stay, history of hypertension, and activity of daily living (ADL) score, white blood cell count (WBC), arterial partial pressure of oxygen (PaO2), prothrombin time (PT), fibrinogen (FIB) on the day of first SICU discharge. Self-help sampling test was carried out on the Nomogram map, and the results showed that the coherence index (C-index) was 0.962 [95% confidence interval (95%CI) = 0.869-1.057]. The classification performance of the model was good. Conclusions The common pathogenies of SICU readmission for patients who underwent hepatobiliary-pancreatic surgery were infection, heart failure, infarction and bleeding. Risk factors of readmission after SICU discharge included the length of first SICU stay, history of hypertension, and ADL score, WBC, PaO2, PT, FIB on the day of first SICU discharge. The model consisted of above risk factors showed a good performance in predicting the probability of readmission after SICU discharge for patients who underwent hepatobiliary-pancreatic surgery.
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Objective To compare the intestinal microbiota of septic shock patients and healthy subjects,and study the composition of the intestinal microbiota and its effect on septic shock patients in the intensive care unit (ICU).Methods A total of 15 stool samples were prospectively collected from septic shock patients admitted to the ICU in the First Affiliated Hospital of Zhengzhou University between June 2015 and February 2016,while 15 samples from healthy subjects served as controls.Bacterial DNA was submitted for 16S rDNA gene sequencing.The association between gut microbiota composition and clinical parameters was evaluated.Shannon index was used to assess the bacterial diversity.Results Compared with the healthy subjects,the composition of intestinal microbiota in septic shock patients changed significantly.The abundance of Proteobacteria and Fusobacteria were significantly higher in septic shock patients than in healthy subjects (23.71％ vs 3.53％,P=0.000 6;1.27％ vs 0.12％,P=0.059,respectively).In this study,29 species were identified,and the composition of intestinal microbiota in each patient was highly individualized.There was no significant difference in Shannon index between septic shock patients and healthy subjects (P=0.12).Conclusions The composition of intestinal microbiota in septic shock patients was characterized by high diversity and individualization,but there was the phenomenon of overproduction of single bacteria genus.The relationship between the composition of intestinal microbiota and clinical outcomes requires further exploration by large sample studies.
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Objective To investigate the value of continuous renal replacement therapy (CRRT) coupled with minimally invasive ultrasound-guided percutaneous transhepatic gallbladder drainage (PTGD) for the treatment of severe acute biliary pancreatitis.Methods Hospitalized patients with severe acute biliary pancreatitis were recruited from the intensive care unit (ICU) of the Mfiliated Hospital of Qingdao University from June 2010 to June 2015,and divided into conventional CRRT alone group (n =30) and CRRT + PTGD group (n =30).Comparisons of postoperatively symptoms (time required for abdominal pain relief,time consumed for,gastrointestinal decompression),laboratory findings (WBC,PLT,PCT,CRP,AMS,TBIL,ALT,ALB,Lac) and acute physiology and chronic health evaluation score (APACHE Ⅱ,Balthazar CT,MODS) were carried out between two groups.The occurrence of complications (ARDS,abdominal infection,bile leakage,abdominal hemorrhage,intestinal injury,catheter translocation,catheter dislocation) was observed.The differences in duration of ventilator support,the length of stay in ICU,and fatality rate were compared between the two groups.Results Compared with the conventional CRRT alone group,the postoperative symptoms were significantly relieved,and time required for abdominal pain relief and time consumed for gastrointestinal decompression were evidently shortened in the CRRT + PTGD group (P ＜ 0.05).There were statistically significant differences in laboratory findings (WBC,PLT,PCT,CRP,AMS,TBIL,ALT) between two groups (P ＜ 0.05).The differences in APACHE Ⅱ,Balthazar CT and MODS score between the two groups also presented statistical significance (P ＜ 0.05).The comparisons of the duration of ventilator support [(6.1 ± 1.3) d vs.(9.5 ± 1.4) d] andthe length of stay [(15.7 ± 1.1) dvs.(21.1 ± 2.5) d] between thetwo groups revealed statistical significance (P ＜ 0.05).Conclusions CRRT coupled with PTGD for the treatment of severe acute biliary pancreatitis can effectively eliminate the inflammatory mediators and toxins from patients.On this basis,the coupled therapy with gallbladder puncture and drainage is capable of decompressing the biliary tract,improving liver function,effectively relieving clinical symptoms,minimizing the changes of laboratory findings an,d APACHE Ⅱ score,and thereby optimizing the prognosis of patients.
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Objective To explore the factors associated with delayed defecation in long-term ventilated patients in intensivecare unit (ICU) and their potential effect on prognosis.Methods A prospective observational cohort study was conducted. The patients admitted to general ICU of the Affiliated Hospital of Qingdao University from October 1st in 2013 to September 30th in 2015 who underwent mechanical ventilation (MV) for ≥6 days were enrolled, and they were divided into early defecation group ( 0.05). Compared with the early defecation group, late defecation group had less patients with loose stools or watery stool at first time [15.7% (13/83) vs. 33.3% (18/54)], more patients using morphine and the usage of epinephrine/norepinephrine more than 24 hours [48.2% (40/83) vs. 40.7% (22/54), 42.2%(35/83) vs. 29.6% (16/54)], higher the maximum PEEP level [cmH2O (1 cmH2O = 0.098 kPa): 7.9±3.7 vs. 6.7±3.5], lower the minimal systolic blood pressure [mmHg (1 mmHg = 0.133 kPa): 74.8±28.1 vs. 88.9±30.2] and more severe of hypoxemia [PaO2/FiO2 < 150 mmHg, 54.2% (5/83) vs. 44.4% (24/53)], all of which had significant differences (allP < 0.05). Factors found statistical significances by single factor analysis were enrolled in the multiple regression analysis, which showed that PaO2/FiO2 < 150 mmHg and systolic blood pressure < 90 mmHg were independently associated with a delay in defecation in patients undergoing long-term MV [PaO2/FiO2 < 150 mmHg: adjusted hazard rate: 1.415, 95% confidence interval (95%CI) = 1.061-1.590,P = 0.026; systolic blood pressure 70-89 mmHg:HR = 1.461, 95%CI = 1.164-1.788, P = 0.002; systolic blood pressure ≤ 69 mmHg: adjusted hazard rate= 1.273, 95%CI = 1.010-1.587,P = 0.034). ICU mortality, the length of ICU stay, CVC indwelling time, duration of MV, ICU acquired bacterial infection rate, and the incidence of VAP at 7 days of MV in late defecation group were significantly higher than those of early defecation group [ICU mortality rate: 15.7% (13/83) vs. 7.4% (4/54), the length of ICU stay (day): 17.0 (14.0, 23.0) vs. 15.0 (13.8, 20.0), CVC indwelling time (days): 12.0 (10.0, 14.0) vs. 10.0 (9.0, 11.3), duration of MV (days): 14.0 (10.0, 20.0) vs. 11.0 (9.8, 15.3), ICU acquired bacterial infections rate: 60.2% (50/83) vs. 14.8% (8/54), the incidence of VAP: 32.5% (27/83) vs. 14.8% (8/54); allP < 0.05]. There was no significant difference in LOD score between both groups. The LOD scores at 4 days and 9 days of MV in late defecation group were significantly higher than those of early defecation group (6.41±4.37 vs. 5.21±3.12, 4.33±2.20 vs. 3.50±2.90, bothP < 0.01).Conclusions PaO2/FiO2 < 150 mmHg and systolic blood pressure < 90 mmHg during the first 5 days of MV were independently associated with a delay in defecation in patients undergoing long-term MV. The results suggest that constipation is associated with adverse outcomes in long-term ventilated patients.
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BACKGROUND:Erythropoietin and bone marrow mesenchymal stem cells have been shown to affect myocardial apoptosis. However, few studies concerned their combined application to sepsis-related myocardial injury. OBJECTIVE:To observe the effects of the combination of erythropoietin and bone marrow mesenchymal stem cells on pathology and apoptosis of sepsis rat cardiomyocytes. METHODS:A total of 50 Sprague-Dawley rats were randomly selected and assigned to five groups (n=10). Sepsis models were established by cecal ligation perforation method. Rat models in the bone marrow mesenchymal stem cellgroup, erythropoietin group and erythropoietin+bone marrow mesenchymal stem cellgroup were respectively treated with bone marrow mesenchymal stem cells, erythropoietin and erythropoietin+bone marrow mesenchymal stem cells immediately after model induction via intraperitoneal injection or caudal vein. Model group received cecum ligation and puncture. Control group did not undergo any treatment after the abdomen was opened. Model and control groups were infused with an equal volume of physiological saline via caudal vein. At 24 hours, experimental animals were sacrificed by anesthesia. Myocardial specimens were col ected. Myocardial appearance was observed using hematoxylin-eosin staining. Bax, Caspase-3 and Bcl-2 were tested by western blot assay. RESULTS AND CONCLUSION:Hematoxylin-eosin staining:cardiomyocytes were regularly arranged, showing integrated structure in the control group. Extensive myocardial fiber breakage, disordered arrangement, cardiomyocyte swel ing or shrinkage, and vacuolar degeneration were observed in the model group. Moreover, myocardial interstitial vascular congestion, edema, and inflammatory cellinfiltration were visible. Myocardial tissue was similar between erythropoietin and bone marrow mesenchymal stem cellgroups, with the presence of mild inflammatory cellinfiltration and scattered normal cardiomyocytes. In the erythropoietin+bone marrow mesenchymal stem cellgroup, cardiomyocytes were slightly damaged. Interstitial vascular congestion was not apparent, and a few inflammatory cells infiltrated. Western blot assay results demonstrated that Bcl-2 protein expression was significantly higher (P<0.01), but Bax and Caspase-3 protein expression was lower (P<0.05) in the erythropoietin+bone marrow mesenchymal stem cellgroup than in the erythropoietin, model and control groups. The combination of erythropoietin and bone marrow mesenchymal stem cells in treatment of sepsis-related myocardial injury could lessen myocardial pathological changes, and inhibit cardiomyocyte apoptosis. The mechanisms maybe exert by upregulating anti-apoptotic protein expression and downregulating apoptotic protein expression.