Objective To investigate the effects of evodiamine on autophagy of human colon a cleno carcinoma lovo cells, and to explore the role and mechanism of autophagy which was induced by evodiamine (EVO). Methods MTT assay combined with the morphologic changes were used to observe the cell viability. Monodansylcadaverine was used to detect autophagy by fluorospectrophotometer and the confocal laser fluorescence microscopy respectively. Immunoblotting assay was used to observe the microtubule-associated protein 1 light chain 3. Finally, evodiamine combined with 3-methyladenine to detect the cell viability with MTT assay and the apoptosis with the flow cytometry, respectively.Results Evodiamine inhibited the viability of Lovo cells in dose-dependent manner ( P ＜ 0. 05 ), especially in 60 μmol/L that was obviously(60% ). Further more, the cell lysis and cell gap widened was observed by the light microscope. Evo triggered the autophagy, and after inhibition the autophagy by 3-MA, the killing capacities of the Evo was enhanced ( P ＜ 0. 01 ). However, autophagy prohibited the apoptosis pathways.Conclusions Evodiamine can trigger the autophagy, which might play a self-defense role in evodiamineinduced cell death. The cytototoxicity of evodiamine can be augmented by the autophagy inhibitors. The joint application of autophagy regulators with the chemotherapeutic agents might enhance the cell killing effects of chemotherapeutic drugs and show a potent role in cancer drug resistance.

Objective To investigate the protective effect of agmatine (AGM) against peritoneal inflammatory response and neutrophil (PMN) infiltration induced by zymosan (ZYM) in mice. Methods Thirty-six adult male C57BL/6 mice were randomly divided into sham group, model group, and AGM treatment group. Peritonitis model was reproduced by intra-peritoneal injection of 1 mg/mL ZYM (0.5 mL), while equivalent phosphate buffer saline (PBS) was given to sham group. 200 mg/kg AGM was injected into peritoneal cavity after ZYM challenge in AGM treatment group. Six mice in each group were sacrificed at 2 hours and 6 hours, respectively, after reproduction of the model. Blood sample and peritoneal lavage fluid (PLF) were collected. The levels of keratinocyte-derived chemokine (KC), macrophage inflammatory protein 2 (MIP-2), tumor necrosis factor-α (TNF-α), interleukins-6 (IL-6) in serum and PLF were determined by enzyme linked immunosorbent assay (ELISA). The number of leukocytes and PMN in PLF were determined by hemocytometer and flow cytometry, respectively. Results Compared with sham group, all serum and PLF levels of KC, MIP-2, TNF-α and IL-6 were greatly elevated at 2 hours after ZYM injection in model group, while AGM treatment could dramatically reduce the levels of the above-mentioned cytokines in serum and PLF as compared with those of the model group [serum KC (ng/L): 990.7±137.9 vs. 2 053.2±262.7, MIP-2 (ng/L): 642.2±124.4 vs. 1 369.7±146.5, TNF-α (ng/L): 608.6±38.1 vs. 1 044.7±101.0, IL-6 (ng/L): 1 058.2±129.1 vs. 1 443.3±190.1; PLF KC (ng/L): 7 462.3±839.6 vs. 12 723.5±1 515.7, MIP-2 (ng/L): 1 570.8±193.4 vs. 3 471.4±384.7, TNF-α (ng/L): 1 115.8±156.7 vs. 1 499.2±231.2, IL-6 (ng/L): 2 646.5±223.2 vs. 3 126.7±291.4; all P < 0.05]. The expressions of KC, MIP-2 and TNF-α at 6 hours were significantly lower than those at 2 hours in model group and AGM treatment group, but IL-6 levels were further increased. The levels of KC and MIP-2 in serum and PLF at 6 hours were decreased to the levels of sham group. At 6 hours after the reproduction of the model, the number of total inflammatory cells and PMN of PLF in the model group was significantly higher than those of the sham group. In contrast, AGM notably lowered the number of inflammatory cells and PMN in peritoneal fluid after ZYM attack [total inflammatory cells (×109/L): 14.7±1.1 vs. 2.0±0.4, 10.1±1.2 vs. 14.7±1.1; PMN (×109/L): 11.37±1.22 vs. 0.18±0.05, 7.69±0.57 vs. 11.37±1.22, all P < 0.05]. Conclusion AGM can effectively alleviate acute peritoneal inflammatory injury induced by ZYM, mainly through reducing the secretion of inflammatory mediators and chemokines, and inhibiting the infiltration of leukocytes and neutrophils.

Objective To observe the effect of agmatine (AGM) on inflammatory factor in Kupffer cells of liver,and to investigate the protective effects of AGM on severe trauma-induced liver injury in mice and its possible mechanism.Methods Forty-two adult male BALB/c mice were randomly divided into sham group,model group,and AGM treatment group,with 14 mice in each group.The mice model of trauma-hemorrhage was reproduced by hindlimbs fracture combined with 35％ of orbital bleeding.The mice in the sham group were only anesthetized without other treatments.The mice in AGM treatment group were given intraperitoneal injection of 200 mg/kg AGM when limited recovery was performed,and the mice in model group were given the equal amount of normal saline.Seven mice in each group were sacrificed at 12 hours and 24 hours,respectively,after modeling,and blood samples and liver tissue were harvested,and liver Kupffer cells were isolated.Serum alanine aminotransferase (ALT),aspartate transaminase (AST)and lactic dehydrogenase (LDH) were determined with automatic biochemistry analyzer.Hepatic pathological changes were observed with light microscope using hematoxylin and eosin (HE) staining.The levels of tumor necrosis factor-α(TNF-o) and interleukin-6 (IL-6) in serum,hepatic homogenate and Kupffer cell supernatant were determined with enzyme linked immunosorbent assay (ELISA).The mRNA expressions of pro-inflammatory cytokines TNF-α and IL-6 in the Kupffer cell were determined by real-time fluorescent quantitation reverse transcription-polymerase chain reaction (RT-qPCR).Results ① The normal liver tissue structure was found in sham group.At 24 hours after modeling in the model group,the changes in pathobiology were found as following:neutrophil infiltration,hepatocytes swelling,hyperemia,and necrosis,as well as the abnormality of parameters reflecting liver function.AGM could significantly improve the pathological changes in liver tissue caused by severe trauma,and ameliorate the liver function.② There were no significant differences in the levels of TNF-α and IL-6 in serum and hepatic tissue at 12 hours after modeling,and the parameters at 24 hours in model group were higher than those at 12 hours,which were significantly higher than those of the sham group [serum TNF-α (ng/L):80.8±4.7 vs.34.7±4.7,IL-6 (ng/L):104.0±9.0 vs.55.4±3.3;liver TNF-α (ng/mg):405.2± 19.6 vs.57.2±10.0,IL-6 (ng/mg):58.4±7.7 vs.14.3±2.1,all P ＜ 0.01].AGM could effectively reduce the levels of TNF-o and IL-6 in serum and hepatic tissue [serum TNF-α (ng/L):58.2 ± 3.1 vs.80.8 ± 4.7,IL-6 (ng/L):74.1 ± 6.6 vs.104.0± 9.0;liver TNF-α (ng/mg):248.7 ± 22.5 vs.405.2 ± 19.6,IL-6 (ng/mg):22.5 ± 3.1 vs.58.4 ± 7.7,all P ＜ 0.01].③ The levels of TNF-o and IL-6 in Kupffer cells supernatant were significantly higher than those of the sham group,and they were further increased after lipopolysaccharide (LPS) stimulation for 24 hours.AGM could effectively reduce the levels of TNF-α and IL-6 in Kupffer cells [TNF-α (ng/L):256.6 ± 5.6 vs.465.5 ± 5.2,IL-6 (ng/L):1 185.5 ± 64.4 vs.2 018.8 ± 53.2,both P ＜ 0.01],and also decreased the mRNA expressions of TNF-α and IL-6 [TNF-α mRNA (2-△△Ct):7.2±0.4 vs.13.5±0.4,IL-6 mRNA (2-△△Ct):13.2±0.7 vs.21.3 ± 1.6,both P ＜ 0.01].Conclusion Agmatine can reduce trauma-induced acute hepatic injury via suppression of cytokines release in Kupffer cells,and can ameliorate the liver function.

ObjectiveTo observe protective effects of agmatine (AGM) on inflammatory response and spleen immune function in mice with trauma.Methods Forty-eight adult male C57BL/6 mice were randomly divided into three groups (n= 16 each), including control group, model group (bilateral femoral fracture and removal of 35% of the total blood volume), and AGM group (trauma/hemorrhage & AGM 200 mg/kg). Eight mice in each group were sacrificed at 3 hours and 24 hours, respectively, after modeling, and blood samples and tissue homogenate of spleen and liver were collected. The contents of tumor necrosis factor-α (TNF-α), interleukins (IL-6, IL-1β) in serum and liver tissue were determined with enzyme linked immunosorbent assay (ELISA). Serum aspartate transaminase (AST), alanine aminotransferase (ALT) and lactic dehydrogenase (LDH) were determined with automatic biochemistry analyzer. Spleen proliferation response stimulated with concanavalin A (ConA) was evaluated with methyl thiazolyl tetrazolium colourimetry (MTT).γ-interferon (IFN-γ) and IL-2 releases were determined with ELISA.Results Compared with control group, 3 hours after trauma/hemorrhage, the levels of serum TNF-α, IL-6, and IL-1β in model group were significantly elevated [TNF-α (ng/L): 145.38±31.50 vs. 23.06±11.14, IL-6 (ng/L): 496.94±50.76 vs. 47.13±17.47, IL-1β (ng/L): 321.31±43.02 vs. 29.25±16.24,allP< 0.01]. It was found that AGM treatment could alleviate the increase in serum pro-inflammatory mediators induced by trauma/hemorrhage, such as TNF-α (ng/L:111.56±25.47 vs. 145.38±31.50), IL-6 (ng/L: 412.56±44.33 vs. 496.94±50.76), IL-1β (ng/L: 273.38±45.25 vs. 321.31±43.02,P< 0.05 orP< 0.01). Twenty-four hours after trauma/hemorrhage, serum pro-inflammatory mediators were recovered to the levels in control group. There was no significant difference in TNF-α and IL-6 levels at 3 hours after trauma/hemorrhage among groups. Compared with control group, the expressions of liver TNF-α and IL-6 in model group were increased at 24 hours following trauma [TNF-α (ng/mg): 32.93±4.90 vs. 26.58±2.33, IL-6 (ng/mg): 11.20±1.66 vs. 8.38±0.89,bothP< 0.01]. However, AGM inhibited the level of TNF-α (ng/mg:28.92±3.16 vs. 32.93±4.90) and IL-6 (ng/mg: 9.03±1.28 vs. 11.20±1.66) in the liver as induced by trauma/hemorrhage (P< 0.05 andP< 0.01). At 24 hours after modeling, model group and AGM group had distinctly higher serum AST, ALT, LDH levels than those of control group [AST (U/L): 405.9±31.2, 245.7±22.1 vs. 128.2±15.9; ALT (U/L): 92.1±6.3, 51.6±5.0 vs. 30.1±3.2; LDH (U/L): 606.7±36.3, 478.7±25.3 vs. 384.0±16.6, allP< 0.01]. Nevertheless,the increase in serum AST, ALT and LDH was alleviated in AGM group (allP< 0.01). Meantime, trauma/hemorrhage produced a noticeable depression of proliferation of splenic cells and IFN-γ and IL-2 release stimulated with ConA compared with control group [proliferation rate: (40.97±4.13)% vs. (89.99±7.76)%, IFN-γ(ng/L): 91.6±12.3 vs. 353.2±21.5,IL-2 (ng/L): 53.4±6.4 vs. 91.0±12.2,allP< 0.01]. In contrast, AGM notably restored the capacity of proliferation response of splenic cells [proliferation rate: (74.86±5.75)% vs. (40.97±4.13)%, P< 0.01],enhanced the release of IFN-γ and IL-2 stimulated with ConA [IFN-γ (ng/L): 327.8±23.6 vs. 91.6±12.3, IL-2 (ng/L): 74.8±10.4 vs. 53.4±6.4, bothP< 0.01].Conclusion AGM can dramatically alleviate spleen immunosuppression, excessive inflammation and organ damage induced by trauma/hemorrhage.

Objective To observe the effects of Yishen Tongluo Decoction (YTD) on the renal mRNA expression of transforming growth factor-β1 ( TGF-β1) and collagen Ⅳ ( ColⅣ) in membranous nephropathy ( MN) rats. Methods SD rats were randomly divided into normal group, model group, benazepril group (in the dosage of 10 mg·kg-1·d-1) , YTD group ( in the dosage of 20 g·kg-1·d-1) . The rats in various groups were given intragastric administration of corresponding agents. At the end of the fourth week, 24-hour urinary protein quantity, albumin ( ALB) , total protein ( TP) , triglyceride ( TG) , total cholesterol ( TC) , blood urea nitrogen ( BUN) , and creatinine (Cr) levels were observed. The mRNA expression levels of TGF-β1 and ColⅣ in renal tissue of rats were detected by immunofluorescence method, electron microscopy and real-time polymerase chain reaction (PCR) method. Results In the model group, urinary protein quantity in rats was increased, serum levels of TP and ALB were significantly lowered, serum levels of TC and TG were significantly increased, renal pathological changes were present, and mRNA expression levels of TGF-β1 and ColIV in renal tissue were up-regulated (P<0.05 compared with normal group). Compared with the model group, 24-hour urinary protein quantity, TC and TG levels were significantly lowered, TP and ALB levels were significantly increased, rat renal injury was relieved, mRNA expression levels of TGF-β1 and ColIV in renal tissue were down-regulated in the treatment groups ( P<0.05) . However, the differences between benazepril group and YTD group were insignificant ( P>0.05) . Conclusion The therapeutic mechanism of YTD for MN is probably related with the inhibition of mRNA expression of TGF-β1 and ColⅣin renal tissue.

Objective To explore the value of reactive hyperemia index (RHI) in predicting the postoperative angina pectoris (AP) in the patients with percutaneous coronary intervention (PCI).Methods Three hundreds and forty-seven patients with coronary heart disease treated by PCI therapy were continuously enrolled in our department from October 10 2015 to August 10 2016.RHI was detected in all cases during hospitalization period by using the noninvasive endothelial function test (Endo-PAT) technique.Then the cases were divided into the control group (RHI≥1.67) and observation group (RHI ＜1.67) according to RHI results.The incidence of AP after discharge from hospital,rehospitalization rate due to AP,frequency and duration of AP were observed in the two groups.Results The AP incidence rates in the control and observation group were 17.04％ and 31.13％ respectively,the difference was statistically significant(P=0.003);the re-hospitalization rate due to AP was 2.22％ and 7.55％ respectively,the difference was statistically significant(P =0.035);the incidence rates of AP attack≥5 times were 6.67 ％ and 16.51 ％ respectively(P=0.008);the duration of AP was (6.39±2.68) min and (8.67±2.58) min,respectively(P=0.001);the Logistic regression analysis showed that the Syntax scores≥23 points and RHI＜1.67 were the risk factors for AP recurrence after PCI(OR=2.265,95％CI:1.354-3.787,P=0.002;OR=2.110,95％CI:1.228-3.628,P=0.007).Conclusion Reduced RHI is closely related with recurrent AP after PCI,increases the incidence,rehospitalization rate due to AP,attack frequency and duration of AP.

Objective:To analyze the application effect of cluster management measures in improving the quality of emergency medical treatment.Methods:By analyzing the problems existing in the work of emergency department, the cluster management scheme was formulated and the intervention measures were implemented from the aspects of intelligent information system, patient management system and medical service process. The accuracy and efficiency of emergency triage, the satisfaction of patients and medical staff, the incidence of medical complaints and disputes and the rate of sudden death were compared before and after cluster management.Results:Before and after the implementation of cluster management, the accuracy of triage classification was 95.0% and 98.7% respectively, and the triage time was (68.3±12.8) s and (50.5±7.2) s respectively( P<0.001). The satisfaction of patients, doctors and nurses increased, the number of complaints decreased from 15 to 5 in half a year, and the number of sudden death decreased from 39 to 23 with a significant difference( P<0.05). Conclusions:The application of cluster management measures in emergency management can improve the medical quality, the satisfaction of medical staff and patients, and ensure the safety of patients.

Objective To explore the risk factors of multiple organ dysfunction syndrome (MODS) in severe trauma patients, put forward a new warning scoring system of MODS, and to provide a more accurate scoring method for doctors to judge the clinical condition and prognosis of patients. Methods Clinical data of 342 patients with severe trauma admitted to intensive care unit (ICU) of the Affiliated Hospital of Zunyi Medical College and Daping Hospital of the Third Military Medical University from January 1st, 2015 to December 31st, 2016 were retrospectively analyzed. The patients were divided into MODS groups (n = 251) and non-MODS group (n = 91) according to clinical outcomes. The clinical data of patients, including gender, age, heart rate (HR) and blood pressure within 24 hours after admission to the hospital, indicators of blood routine and blood biochemistry, severity of disease, severity of trauma, whether received the emergency intubation or surgery within 24 hours or not, whether developed sepsis or acute respiratory distress syndrome (ARDS) during hospitalization, were recorded, and univariate analysis was conducted. The indicators with statistical significance found by univariate analysis were enrolled in multivariate Logistic regression analysis, and the risk factors for MODS in patients with severe trauma were screened and assigned, and the final total score was MODS warning score. Receiver operating characteristic (ROC) curve was plotted to evaluate MODS warning score for predicting the occurrence of MODS in patients with severe trauma. Results Compared with non-MODS group, HR, Na+, serum creatinine (SCr), activated partial thromboplastin time (APTT), injury severity score (ISS), new injury severity score (NISS), acute physiology and chronic health evaluation Ⅱ (APACHEⅡ) score and sequential organ failure assessment (SOFA) score in MODS group were significantly increased, pH value, red blood cell (RBC), platelet (PLT), albumin (Alb) and Glasgow coma score (GCS) were remarkably decreased, and multiple injury, the patients with shock at admission, blood transfusion, central venous catheter, emergency intubation and infection were also increased, and more patients suffered from sepsis and ARDS. Multivariate Logistic regression analysis showed that the number of injured places equal or more than 2, shock at admission, APACHEⅡ score ≥ 15, SOFA score ≥ 4 and APTT > 40 s were risk factors for MODS in patients with severe trauma, with total MODS warning score of 7.5. ROC curve analysis showed that the area under ROC curve (AUC) of MODS warning score for predicting MODS in patients with severe trauma was 0.822, which was significantly higher than that of APACHEⅡ score (AUC = 0.698, P < 0.001), APTT (AUC = 0.693, P < 0.001) and SOFA score (AUC = 0.770, P = 0.025). When the cut-off value of MODS warning score was 2.5, the sensitivity was 61.35%, the specificity was 90.11%, and Youden index was 0.515. Conclusions MODS warning score is composed of five factors, including the number of injured places, shock at admission, APACHEⅡ score, SOFA score and APTT, which could be regarded as early warning score system for predicting MODS in patients with severe trauma. MODS warning score can be more comprehensive and timely to assess the possibility of MODS and prognosis of patients with severe trauma, and the prediction result is better than the single use of APTT, APACHEⅡ or SOFA score.

Humans ; Asthma/drug therapy* ; Biological Therapy