Objective To investigate the clinical value of serum endocan and procalcitonin (PCT) in early diagnosis and prognosis evaluation of sepsis.Methods The patients with systemic inflammatory response syndrome (SIRS,n = 26) and sepsis (n = 78) admitted to intensive care unit (ICU) of the Third Hospital of Hebei Medical University from December 2014 to December 2016 were enrolled. According to the severity of disease, the sepsis patients were divided into general sepsis group (n = 20), severe sepsis group (n = 24), and septic shock group (n = 34). The cases were divided into survival group (n = 55) and non-survival group (n = 23) according to 28-day mortality. The serum endocan, PCT, acute physiology and chronic health evaluation Ⅱ (APACHE Ⅱ) score, and sequential organ failure assessment (SOFA) score were recorded when the patients were admitted into ICU. The differences in endocan, PCT, APACHE Ⅱ, SOFA score between SIRS and sepsis groups and within sepsis subgroups were compared. Spearman correlation analysis was used to analyze the correlation between the indexes of sepsis patients. Receiver operation characteristic curve (ROC) was used to evaluate the value of endocan and PCT for the diagnosis and prognosis of sepsis.Results ① Serum endocan, PCT, APACHE Ⅱ, SOFA score and 28-day mortality in the sepsis group were significantly higher than those in the SIRS group [endocan (μg/L): 4.28 (10.64) vs. 1.03 (0.69), PCT (μg/L): 3.94 (10.75) vs. 0.43 (0.39), APACHE Ⅱ:18.81±9.17 vs. 9.35±3.78, SOFA: 9.00 (7.20) vs. 4.50 (1.50), 28-day mortality: 29.49% vs. 11.54%, allP < 0.01]. The area under the ROC curve (AUC) of endocan, PCT, APACHE Ⅱ, SOFA score for sepsis diagnosis were 0.887, 0.842, 0.822, 0.835, respectively. When the cut-off value of endocan was 1.26μg/L, the sepsis diagnostic sensitivity was 87.2% and specificity was 81.8%. When the cut-off value of PCT was 0.75μg/L, the sepsis diagnostic sensitivity was 85.9% and specificity was 81.8%. ② With the severity of the disease increased, the index showed an increasing trend in patients with sepsis. Serum endocan, PCT, APACHE Ⅱ, SOFA score and 28-day mortality in septic shock group were significantly higher than those in severe sepsis group or general sepsis group [endocan (μg/L): 13.02 (6.70) vs. 3.33 (3.05), 1.60 (0.98); PCT (μg/L): 8.10 (17.68) vs. 5.47 (8.92), 1.57 (2.78); APACHE Ⅱ: 25.00 (9.50) vs. 18.00 (9.00), 9.50 (5.75); SOFA: 13.00 (4.50) vs. 8.00 (3.00), 5.00 (3.50); 28-day mortality: 52.94% vs. 20.83%, 0%; allP < 0.01]. There was a significantly positive correlation between endocan, PCT, APACHE Ⅱ, SOFA, indicating that the endocan and PCT can be used to assess the severity of sepsis. ③ Serum endocan, PCT, APACHE Ⅱ and SOFA score in non-survival group were significantly higher than those in the survival group [endocan (μg/L): 15.05 (9.23) vs. 2.32 (4.81), PCT (μg/L):18.40 (16.99) vs. 3.10 (6.67), APACHE Ⅱ: 28.13±7.56 vs. 14.91±6.64, SOFA: 14.70±3.65 vs. 7.38±3.26, allP < 0.01]. The AUC of endocan, PCT, APACHE Ⅱ, SOFA score for the prediction of non-survival sepsis were 0.915, 0.763, 0.899, 0.930. When the cut-off value of endocan was 4.37μg/L, the septic death prediction sensitivity was 95.7% and specificity was 70.9%. When the cut-off value of PCT was 7.68μg/L, the septic death prediction sensitivity was 65.2% and specificity was 78.2%.Conclusions Serum endocan is more clinically valuable than PCT in early diagnosis and prognosis assessment of sepsis.

Objective To evaluate the effects of levosimendan on hemodynamics and cardiac function in patients with septic shock. Methods A prospective single-center randomized controlled trial was conducted. The patients with septic shock admitted to the Department of Critical Care Medicine of the Third Hospital of Hebei Medical University from June 2011 to October 2013 were enrolled. The patients with septic shock received the conventional treatment according to international guidelines for management of severe sepsis and septic shock. Thirty-six patients received the examination of echocardiography and left ventricular ejection fraction (LVEF)≤0.45 after fluid resuscitation were enrolled the study ,who were divided into two groups according to random number table ,with 18 cases in each group. After the conventional treatment,the patients in dobutamine group received intravenous injection of 5μg·kg-1·min-1 dobutamine for 48 hours immediately after fluid resuscitation,and those in levosimendan group received a 24-hour infusion of 5μg·kg-1·min-1 dobutamine followed by a 24-hour infusion of 0.2μg·kg-1·min-1 levosimendan. The hemodynamics and cardiac function were evaluated by pulse indicator continuous cardiac output (PiCCO) and ultrasound during treatment. Results Compared with dobutamine group,after the treatment in the levosimendan group,stroke volume index (SVI),cardiac index (CI)and left ventricular stroke work index(LVSWI) were significantly increased〔SVI(mL/m2):39.8±5.4 vs. 37.5±4.5,t=-2.762,P=0.020;CI(L·min-1·m-2):4.6± 0.7 vs. 3.6±0.7,t=-9.829,P=0.000;LVSWI (kg·min-1·m-2):33.7±2.4 vs. 28.2±1.2,t=-6.307,P=0.000〕, and central venous pressure (CVP),intrathoracic blood volume index (ITBVI)and extravascular lung water index (EVLWI)were significantly decreased〔CVP(mmHg,1 mmHg=0.133 kPa):8.2±0.9 vs. 12.1±0.8,t=3.928,P=0.002;ITBVI (mL/m2):820±42 vs. 978±69,t=9.472,P=0.000;EVLWI (mL/kg):6.1±1.6 vs. 8.9±1.7,t=4.467,P=0.001〕. Cardiac ultrasound showed that compared with dobutamine group,in the levosimendan group,left ventricular end-systolic volume (LVESI) and end-diastolic volume (LVEDI) were significantly lowered〔LVESI (mL/m2):32.7±9.2 vs. 48.2±13.4,t=0.882,P=0.000;LVEDI (mL/m2):61.7±11.4 vs. 78.6±13.6,t=2.453, P=0.032〕,and the LVEF was significantly increased (0.463 ±0.068 vs. 0.383 ±0.085,t=-2.439,P=0.035). Levosimendan also could decrease the lactic acid(mmol/L:3.4±1.1 vs. 5.2±1.2,t=3.346,P=0.007),and increase the lactate clearance rate(mL/min:73.2±13.5 vs. 47.6±11.8,t=-4.079,P=0.002),24-hour urinary output(mL:2 213.4±354.0 vs. 1 533.8±402.0,t=6.342,P=0.000)and 24-hour cumulative intake (mL:5 746.6±420.0 vs. 4 156.7 ±215.0,t=7.126,P=0.000). There were no significant differences in total volume of norepinephrine, mortality in intensive care unit (ICU)and 28-day mortality between two groups. Conclusion Levosimendan can increase cardiac ejection function,reduce the heart blood and vascular preload,intrathoracic lung water,improve heart function and systemic hemodynamic indexes of patients with septic shock.

Objective To evaluate the role of miRNA-214 in myocardial injury in septic mice.Methods Sixty-six healthy male Kunming mice,weighing 25-30 g,were randomLy (random number) divided into4 groups:sham operation group (Sham group,n =18),cecal ligation and puncture group (CLP group,n =18),CLP + miRNA-214 precursor treated group (pre-miR-214 group,n =12),CLP + miRNA-214 inhibitor treated (anti-miR-214 group,n =12),and the rest six mice were treated with miRNA-214 precursor or inhibitor intravenously.Sepsis was induced by CLP,pre-miR-214 group and anti-miR-214 group were respectively treated with miRNA-214 precursor or miRNA-214 inhibitor before the CLP,Sham group were exposed to the cecum only.Mice were sacrificed and hearts were removed for determination of miRNA-214 expression by RT-PCR in Sham and CLP group at 6 h,12 h,and 24 h.Blood samples were collected from inferior vena cava at 12 h and 24 h after CLP for determination of the B-type natriuretic peptide (BNP) and cardiac troponin-I (cTnI) concentration by ELISA,then the mice were sacrificed and hearts were removed for determination of the inflammatory factor concentration by ELISA,cardiac myocyte apoptosis rate by flow cytometry and histopathological changes by microscopic examination.Statistical analysis was carried out with SPSS 13.0 software for One-way ANOVA.Results (1) The miRNA-214 expression was higher in CLP group than that in Sham group at 6 h,12 h and 24 h;(2) Compared with CLP group,the miRNA-214 expression in pre-miR-214 group was increased,but decreased in anti-miR-214 group at 12 h;(3) Compared with CLP group,the concentrations of the serum BNP,cTNI,tumor necrosis factor-alpha (TNF-α) and the interleukin-6 (IL-6) were lower than those in pre-miR-214 group,whereas the interleukin-10 (IL-10) was increased.However,the levels of these variables changed just in opposite direction in anti-miR-214 group at 12 h,24 h;(4) Compared with CLP group,the myocardial cell apoptosis rate was decreased in pre-miR-214 group,but increased in anti-miR-214 group at 24 h;(5) The microscopic examination showed that myocardial injury was attenuated in pre-miR-214 group compared with CLP group.Conclusions The miRNA-214 expression was increased in myocardial injury in CLP mice,suggesting miRNA-214 expression relating to myocardial protection in sepsis.

BACKGROUND: As a kind of polypeptide, angiotensin-converting enzyme (ACE) inhibitory peptide can lower the blood pressure of human body through restraining the formation of angiotensin Ⅱ.OBJECTIVE: To investigate the influence of AGE inhibitory peptide on endothelial cell proliferation and endothelin expression in cultured human umbilical vein cells based on cellular and molecular levels in order to provide the experimental evidences for ACE inhibitory peptide to be the potential blood pressure-lowering health food.DESIGN: Repeated measures design.SETTING: School of Bioscience and Bioengineering, South China University of Technology; School of Applied Chemistry and Biological Technology,Shenzhen Polytechnic.MATERIALS: The experiment was carried out in the Institute of Applied Chemistry and Biological Technology, Shenzhen Polytechnic from September 2004 to March 2005. The AGE inhibitory peptide was provided by the Institute of Applied Chemistry and Biological Technology, Shenzhen Polytechnic. Under certain circumstance, 15 μ mol/L of the inhibitor was needed to decrease half of the AGE activity. The human umbilical vein endothelial cells of the 4th generation were cultured randomly in 7 groups with different concentrations: medium group, 150, 300 and 600 mg/L ACE inhibitory peptide groups, captopril group, norepinephrine(NE) group, and ACE inhibitory peptide+NE group.METHODS: ①The endothelial cells were cultured as recommended. The medium was M199+FBS(0.15, v/v)+penicillin(10 000 U/mL)+streptomycin (100 mg/L). After cellular fusion, the cells were carried on the passage with the ratio of 1:2. The 4th generation cells were used for experiment. ②M199(0.15, v/v) was contained in each group. ACE inhibitory peptides were added to make the final concentration 150, 300 and 600 mg/L in the 150, 300 and 600 mg/L ACE inhibitory peptide groups respectively. Captopril was added to make the final concentration 10-5 mol/L in the captopril group. NE was added to make the final concentration 100 μg/L in the NE group. ACE inhibitory peptide and NE were added to make the final concentration 300 mg/L and 100 μg/L in the ACE inhibitory peptide+NE group respectively. ③The state of cell growth was determined with cytometry. The contents of endothelial cells in the medium with different culture times were determined with radioimmunoassay. The expression of endothe lin mRNA was determined with reverse transcriptase-polymerase chain reaction. The expression of cellular endothelin protein was determined with immunohistochemical method.MAIN OUTCOME MEASURES: ①The influence of ACE inhibitory peptide on endothelial cell proliferation. ②The influence of ACE inhibitory peptide on the endothelin mRNA and endothelin protein.RESULTS: ①The influence of ACE inhibitory peptide on endothelial cell proliferation and endothelin secretion: Compared with the medium group,in the captopril and 150, 300, 600 mg/L ACE inhibitory peptide groups,the growth of endothelial cells was restrained and the endothelin content in the medium was lowered(P ＜ 0.01 or 0.05). NE could promote the growth of endothelial cells and the secretion of endothelin, but the cell density and endothelin content after treatment with ACE inhibitory peptide were similar to those in the medium group (P ＞ 0.05). ②The influence of ACE inhibitory peptide on the expressions of endothelin mRNA and protein in endothelial cells: Compared with the medium group, the expressions of endothelin mRNA and protein might be lowered in the captopril and 150,300, 600 mg/L ACE inhibitory peptide groups(P ＜ 0.01 or 0.05). The expressions of endothelin mRNA and protein could be up-regulated by NE.The gene expression after treatment with ACE inhibitory peptide was similar to that in the medium group(P ＞ 0.05).CONCLUSION: The ACE inhibitory peptides of different dosages can all restrain the growth of endothelial cells, lower the endothelin content, decrease the expression of endothelin gene and resist NE improved growth and secretion of endothelial cells in umbilical vein cell effectively.

Objective To compare the blood high mobility group box-1 (HMGB-l) versus N-terminal prohormone of brain natriuretic peptide (NT-proBNP) for assessment of cardiac dysfunction in the patients with severe sepsis.Methods Ninety-eight patients of both sexes,aged 18-75 yr,were divided into 3 groups according to left ventricular ejection fraction (LVEF):non-cardiac dysfunction group (LVEF ≥ 50 ％,n =43),mild cardiac dysfunction group (30％ ≤ LVEF ＜ 50％,n =41),and severe cardiac dysfunction group (LVEF ＜ 30％,n =14).Acute physiology and chronic health evaluation Ⅱ (APACHE Ⅱ) score was performed.Venous blood samples were obtained for determination of the concentrations of HMGB-1 and NT-proBNP.Days of hospitalization and the survival rates within 28 days after admission to hospital were recorded.Pearson correlation between blood HMGB-l,NT-proBNP concentrations and LVEF was analyzed.Results Compared with non-cardiac dysfunction group,LVEF was significantly decreased,APACHE Ⅱ score and blood HMGB-1 and NT-proBNP concentrations were increased,days of hospitalization were prolonged,and the survival rates were decreased in mild and severe cardiac dysfunction groups,and the changes in the parameters mentioned above were more obvious in severe cardiac dysfunction group.The correlation coefficient between blood HMGB-1 concentration and LVEF was -0.639 and between blood NT-proBNP concentration and LVEF was-0.521,showing significant difference.Conclusion Blood HMGB-1 concentration provides higher reliability than blood NT-proBNP concentration in assessing the degree of cardiac dysfunction in the patients with severe sepsis.

Objective To investigate the protective effects of erythropoietin (EPO) on myocardium injury after sepsis in rats in order to clarify the mechanisms.Methods The rat models of sepsis were produced by cecal ligation and perforation method (CLP).Ninty-six healthy SD rats were randomly (random number) divided into 3 groups:the sham operation group (Sham group,n =32),the sepsis group (CLP group,n =32),and the EPO treatment group (EPO group,n =32) treated with EPO 1000 IU/kg intraperitoneal injection after the CLP.The observation intervals were set at 3 h,6 h,12 h and 24 h after the surgery.The cardiac hemodynamics of the CLP group were measured.Plasma levels of inflammatory factors and myocardial enzyme indicators were determined by enzyme-linked immunosorbent assay (ELISA) ; Membrane potential levels of chondriosome of myocardial cells,cell apoptosis rates and expressions of nuclear factor-κB p65 of myocardium tissue were detected by flow cytometer; Then the pathological change of myocardium with HE staining was observed under light microscopy.Results (1) Compared with the CLP group,left ventricle systolic pressure (LVSP),left ventricle diastolic end pressure (LVEDP),the maximum rate of left ventricle rise and fall (+ dp/dtmax and-dp/dtmax) in the EPO group improved (P ＜0.05,P＜0.01); (2) Compared with the Sham operation group,plasma levels of tumor necrosis factoralpha (TNF-α),interleukin-6 (IL-6),C-reactive protein (CRP),cardiac troponin-I (cTNI),creatine kinase (CK),lactate dehydrogenase (LDH) and glutamine-oxaloacetic transaminase (GOT) in the CLP group increased at each interval (P ＜ 0.05),and those biomarkers in the EPO group were lower than those in the CLP group (P ＜ 0.05) ; On the contrary,plasma level of anti-inflammatory factor IL-10 in EPO group was higher than that in the CLP group (P ＜ 0.01) ; (3) Compared with the Sham operation group,the cell apoptosis rates in the CLP group increased significantly (P ＜ 0.01),and it decreased obviously in the EPO group (P ＜ 0.01); (4) Compared with the Sham group,membrane potential levels of chondriosome of myocardial cell in the CLP group decreased (P ＜0.01),while it increased in the EPO group in comparison with the CLP group (P ＜ 0.01) ; (5) Pathological changes of myocardium after the CLP could be lessened by the EPO treatment.Conclusions EPO may increase membrane potential levels of chondriosome and decrease the apoptosis rates of myocardial cell in rats after sepsis,and it may reduced the production of inflammatory factors to protect the myocardial cell by down-regulating NF-κB p65.Both increased membrane potential level of chonriosome and decreased inflammatory factor may implicate in myocardium protection thereby improving cardiac function after sepsis.

Objective To study the absorption mechanism of the antihypertensive peptide,Val-Leu-Pro-Pro-Val(VLPVP) in Caco-2 cell model. Method Caco-2 cell model was used in vitro to investigate the effects of time,pH,drug concentration,enhancers and inhibitors on the absorption of VLPVP. VLPVP concentration was measured by HPLC. Results The transport rate of VLPVP reached the maximal level at pH7.4,and was positively correlated to transport time and drug concentration. The peptide transporter inhibitors(Gly-Pro,arphamenine A) ,the endocytosis inhibitor(phenyl arsenoxide) didn’t influence the transport of VLPVP,while the paracellular transport enhancer(sodium deoxycholate) ,the inhibitor of multi-drug resistant protein(MK-571) and the energy inhibitor(sodium azide) enhanced it from apical to basolateral side significantly,but the latter two didn’t influence the transport from basolateral to apical side. Conclusion Paracellular diffusion was suggested to have been the main mechanism for the absorption of intact VLPVP and MRP2 had strong eflux effect on the transepithelial transport of VLPVP.

Objective To study the effect of angiotensin converting enzyme inhibitory peptides (ACEIP) on blood pressure of spontaneously hypertensive rats.Methods One hundred and four spontaneously hypertensive rats(SHR) were randomized to thirteen groups (n= 8,each group) and received one of following approaches:saline,captopril (10 mg/kg),low-dose (200 ?g/kg),medium-dose(400 ?g/kg) and high-dose (800 ?g/kg) of the three ACEIPs:VLPVP,KVLPVP,VLPVPR respectively.Thirty-two WKY rats were randomized to 4 groups(n=8 in each group),VLPVP,KVLPVP,VLPVPR (800 ?g/kg by gavage) and saline were administrated.The systolic blood pressure was measured before and after the administration of ACEIP.Results Compared with the control group,medium-dose and high-dose of VLPVP,KVLPVP and VLPVPR reduce blood pressure in hypertensive rats and the effect can last for above 6 hours [decline of blood pressure:high-dose:(40.9?15.7),(38.0?21.3) and (43.2?12.6) mmHg;medium-dose:(28.3?23.0),(24.7?21.5) and (26.2?9.9);control:(8.5?5.1),(6.2?4.1) and (7.1?5.5)mmHg;all P0.05].Conclusion Recombinant ACEIP acutely reduce high blood pressure in SHR,the antihypertensive effects persist for at least 6 hours,with no effect on normotensive WKY rats.

Objective To study the effects of the antiarrhythmic drugs of propafenone, amiodarone and dilthiazem on c-type Kv1. 4 channels in Xenopus laevis oocytes with two-electrode voltage-clamp technique. Methods Defolliculated oocytes ( stage Ⅴ - Ⅵ) had transcribed cRNAs of ferret Kv1. 4△N channels injected. The oocytes were continuously perfused with control solution or propafenone, amiodarone and dilthiazem under monitoring of software of Clampfit v 9. 0. Results All of the three drugs blocked ferret Kv1. 4△N channel in voltage-, frequency- and concentration-dependent manners. The values of IC50 (50%inhibiting concentration ) of propafenone, amiodarone and dilthiazem were ( 103.4± 2. 2 ) μ mol/L,(501.22 +5.9) μmol/L and (353.62 +9.9) μmol/L, respectively. The currents under the actions of propafenone , amiodarone and dilthiazem were decreased to 41%, 32% and 21% of control group, respectively. Propafenone (100μ mol/L), Amiodarone (500μmol/L), dilthiazem (350μmol/L) inhibited currents to (54. 6 + 1.9 ) %, ( 46. 3 + 3.5 ) %, ( 52. 8 ± 2. 8 ) % of control group in voltage-dependent blockage. Conclusions The results suggested that all the three drugs blocked the kv1. 4△N channel in the open state. The three drugs block the kv1. 4△N channel maybe had the similiar effects in some respects, but each had its unique characteristics.

Objective and Method: To study the influence of angiotensin converting enzyme inhibitory peptides (ACEIP) on the expression of eNOS, iNOS, ET-1 mRNA in cultured human umbilical vein endothelial cells by semi-quantitative reverse transcription-polymerase chain reaction(RT-PCR) method and then investigate its mechanism. Results: Campared with the control, the expression of ET-1 and iNOS was lower and the expression of eNOS was higer in different ACEIP groups. Conclusion: The antihyertensive function of ACEIP partly depended on the its effect to lessen the expression of ET-1,iNOS and induce the expression of eNOS in cultured human umbilical vascular endothelial cells.