Objective To investigate the effect of gradient heat stress on phagocytosis of hepatic Kupffer cells (KCs) in vitro in rats. Methods Rat Kupffer cells were isolated in vitro and the temperature for gradient heat stress was set at 37, 39, 41 and 43℃. After thermal stimulation, cell injury was detected by PI and Hochest33342 staining. CCK-8 assay was used to investigate difference in cellular proliferation rate over 24h between the groups. Flow cytometry was used to investigate the influence of heat stress on the phagocytosis of KCs. Results Compared to the normal control group, cells in each heat stress group exhibited varying degrees of damage, especially cells in 43℃ group. The ratio of damage cells increased with the increase of heat stress severity (P<0.05). Proliferation assay indicated that the proliferation rate of cells in each heat stress group was significantly decreased in comparison with normal control group 6h after heat stress (P<0.05). After 12h recovery, decrease in proliferation rate was observed only in 43℃ group (P<0.001), and no difference in the rate of proliferation could be observed between the heat stress groups and normal control group after 24h recovery. Flow cytometry showed, that the phagocytosis of KCs decreased in heat stress groups compared with control group, especially in 43℃ group (P<0.05). This phenomenon disappeared after 24h recovery. Conclusion Heat stress can inhibit the phagocytosis of rat liver KCs through its cytotoxic effect on KCs, and subsequently inhibits its proliferative ability. Further investigation of the effect of heat stress on KCs may help understand the pathogenesis of heat stress.

Objective To investigate the effects of large dose of atorvastatin on the expression of Sprouty-1 in CD4 + T lymphocytes from unstable angina patients during perioperative period of PCI.Methods A total of 52 unstable angina patients enrolled were divided randomly (random number) into large-dose atorvastatin (80 mg/d,n =26) pretreated group and moderate-dose atorvastatin (20 mg/d,n =26) pretreated group.Circulating CD4 + T cells were obtained by magnetic cell sorting system (MACS) before PCI and 18-24h after PCI.For detecting the gene expression,the reverse transcription fluorescent quantitative polymerase chain reaction (RT-PCR) was used to measure the expression of Sprouty-1 mRNA in CD4 + T lymphocyte.The level of Sprouty-1 protein was detected with Western blot analysis and IL-2 was quantified by enzyme-linked immunosorbent assay (ELISA).Results ①Compared with large-dose group before PCI,the expression of Sprouty-1 mRNA and Sprouty-1 protein levels were dramatically increased in large-dose group after PCI (P ＜ 0.05).②Compared with moderate-dose group before PCI,the expression of Sprouty-1 mRNA and protein levels were slightly increased in moderate-dose group after PCI,but there was no statistical significance (P ＞ 0.05).③Compared with large-dose group before PCI,the serum level of IL-2 was decreased in large-dose group after PCI (P ＜ 0.05).Whereas the serum level of IL-2 was slightly increased in moderate-dose group after PCI compared to moderate-dose group before PCI,there was still no statistical significance (P ＞ 0.05).Conclusions Large-dose atorvastatin pretreatment reduced post-PCI myocardial inflammation through up-regulating the expression of Sprouty-1 mRNA and level of Sprouty-1 protein in CD4 + T lymphocytes.

Objective To investigate the effects of aggressive dosing of atorvastatin on the expression of SOCS1 in CD4 + Tlymphocytes from patients with unstable angina pectoris during peri-operative period of PCI.Methods A cohort of 50 patients with unstable angina pectoris were randomized (random number) to give pretreatment with either an aggressive dose (80 mg/d,n =25) or a routine dose (20 mg/d,n =25)of atorvastatin.Circulating CD4 +T cells were subsequently obtained prior to PCI,and also 18 h to 24 hours after PCI,using a magnetic cell sorting system (MACS).Fluorescence-based quantitative real-time polymerase chain reaction (qRT-PCR) was used to measure expressions of SOCSI mRNA in the isolated CD4 + Tlymphocytes,and western blot analysis was used to detect levels of SOCS1 protein.Serum levels of IFN-γwere quantified using enzyme-linked immunosorbent assays (ELISAs).Results Compared with routine dose group,the expressions of SOCS1 mRNA and protein levels were dramatically increased and those were higher in aggressive dose group following PCI (P ＜ 0.05).In contrast,serum levels of IFN-γsignificantly increased following PCI in both groups,but it was higher in routine dose group than in aggressive dose group (P ＜ 0.05).Conclusions Treatment with aggressive dosing of atorvastatin reduced the post-PCI myocardial inflammatory response in patients with unstable angina pectoris,possibly modulating by up-regulating SOCS1 expression in CD4 + Tlymphocytes.

BACKGROUND: Coronary microembolization (CME) is a serious complication following percutaneous coronary intervention (PCI) in patients with acute coronary syndromes. The use of metoprolol before PCI can significantly protect ischemic myocardium from myocardial damage, but the function of metoprolol in the treatment of CME is not entirely clear. This study was to explore the effect and significance of metoprolol on myocardial apoptosis and caspase-3 activation after CME in rats. METHODS: Thirty rats were randomly divided into three groups including sham-operation (control group), CME plus saline (CME group), CME plus metoprolol (metoprolol group), 10 rats for each group. The CME group was induced by injecting 3000 polyethylene microspheres (42 μm) into the left ventricle during a 10-second occlusion of the ascending aorta; the control group was injected with physiological saline instead of microembolization ball; the metoprolol or saline group was given three intravenous bolus injections before CME. Echocardiography, TUNEL staining, and Western blotting were used to evaluate cardiac function, proportion of apoptotic cells and activation of caspase-3 respectively at 6 hours after operation. RESULTS: Echocardiographic parameters displayed that the metoprolol group improved cardiac function significantly compared with the CME group (P<0.05). The myocardial apoptotic rate of the CME group as wel as the contents of activated caspase-3 increased significantly (P<0.05), both of which were ameliorated significantly by metoprolol treatment (P<0.05). CONCLUSIONS: This study demonstrates that metoprolol can protect the myocardium during CME in rats by inhibiting apoptosis and improving cardiac function. These results suggest that the inhibition of apoptosis can be a potential therapeutic strategy for the treatment of CME.

Objective:Using lasso regression analysis to screen out the blood lipid indexes closely related to coronary heart diseaseMethods:The clinical data of 3 062 patients with coronary heart disease who were hospitalized in the Department of Cardiology, Affiliated Hospital of Jining Medical College from May 2013 to November 2015 were retrospectively analyzed.They were divided into control group ( n=2 427) and coronary angiography group ( n=635). R language was used for statistical analysis.Multiple logistic regression models were established for indicators of blood lipid related to CAD, and their multicollinearity severity was assessed.LASSO regression was used to screen out the representative lipid parameters in the CAD prediction model. Results:A total of 3 062 patients were enrolled, including 2 427 patients in coronary heart disease group and 635 patients in control group.The inclusion of lipid parameters into multiple logistic regression model leads to serious multicollinearity.Stepwise regression can only partially reduce multicollinearity severity, while LASSO regression model significantly reduces multicollinearity severity.Low density lipoprotein cholesterol (LDL-C), high density lipoprotein cholesterol (HDL-C) and non-high density lipoprotein cholesterol (non-HDL-C) were found to be the representative lipid indexes for predicting coronary heart disease by LASSO regression analysis.Conclusion:LASSO regression has advantages in processing multicollinearity data.LASSO regression showed that LDL-C, HDL-C and non-HDL-C were representative lipid indicators for predicting coronary heart disease..

Objective To investigate the effects of ultrasound-targeted microbubble destructionmediated MicroRNA-21 on cardiomyocyte apoptosis after coronary microembolization (CME) in swine.Methods Twenty Bama miniature swine were randomLy (random number) divided into sham-operated,CME,CME plus gene transfection and CME plus ultrasound mediated gene transfection groups (n =5 per group).The CME model was established by microcatheter-mediated injection of microspheres into the left anterior descending artery.The sham-operated group were made by injection of saline instead.The CME plus ultrasound mediated gene transfection group was made by injection of plasmid-microbubble mixture through the marginal ear vein 4 days before CME established.Meanwhile,ultrasound treatment was given to the myocardium through chest wall.The CME plus gene transfection group was made by injection of plasmidmicrobubble mixture through the marginal ear vein 4 days before CME established without exposure to ultrasound.Left ventricular ejection fraction (LVEF) was examined by cardiac ultrasound.Tissue biopsy was stained with hematoxylin-eosin (HE) and hematoxylin basic fuchsin picric acid (HBFP) to measure the size of infarction area.Green fluorescent protein (GFP)-labeled gene expression was evaluated by fluorescent microscopy in frozen sections.Cardiomyocyte apoptosis was detected with terminal deoxynucleotidyl transferase (TdT)-mediated dUTP nick end-labeling (TUNEL staining).The expression of PTEN mRNA was measured by fluorescent quantitative PCR.The levels of PTEN protein and Caspase-3 protein was measured by western blot.Results ①Compared to CME plus gene transfection group,the CME plus ultrasound mediated gene group had over eightfold expression of exogenous genes in myocardium (P ＜ 0.05) measured by using optical density of green fluorescence protein;② Compared with shamoperated group [(67.87 ±2.36)％],the LVEF of CME group [(50.94 ±3.52)％] and CME plus gene transfection group [(52.47 ±3.71)％] were markedly decreased (P ＜ 0.05).Compared with CME group,the CME plus ultrasound mediated gene transfection group [(64.79 ± 2.95)％] improved CME-induced cardiac dysfunction as evidenced by increased LVEF (P ＜ 0.05);③Compared with sham-operated group,the expression of PTEN mRNA and levels of PTEN protein and Caspase-3 protein in the CME group increased significantly (P ＜ 0.05).Compared with CME group,the levels of PTEN protein and Caspase-3 protein and the expression of PTEN mRNA in CME plus ultrasound mediated gene transfection group was dramatically decreased (P ＜ 0.05).Conclusions Ultrasound microbubble-mediated MicroRNA-21 transfection effectively improved CME-induced cardiac dysfunction by down-regulating the expression of targeted gene PTEN in myocardial cells,mainly reducing the post-CME myocardial cell apoptosis.

Objective To investigate the effect of Aidi injection combined with chemotherapy on CD+4CDhi25CDlow127regulatory T (Treg) cells in peripheral blood of advanced non-small-cell lung cancer (NSCLC).Methods Sixty patients with advanced NSCLC were randomly divided into the study group (treated with Aidi injection combined with chemotherapy) and the control group (treated with chemotherapy).The levels of CD+4CDhi25CDlow127Treg cells were detected by flow cytometry (FCM), and TGF-β1 and IL-10 levels were determined by enzyme linked immunosorbent assay (ELISA) before and after chemotherapy. A group consisted of 20 healthy persons was set up meanwhile.Results There was a significantly higher percentage of CD+4CDhi25CDlow127Treg cells in patients with NSCLC (5.77±1.50) % than that(3.84±0.96) % of healthy volunteers (P=0.000).The IL-10 and TGF-β1 levels[(24.09±6.74), (197.76±43.76) ng/ml]in the serum of patients with NSCLC were also significantly higher than that[(19.39±5.73), (141.13±32.17) ng/ml]of healthy volunteers (P =0.006,P =0.002). In control group, levels of CD+4CDhi25CDlow127 Treg cells were obviously lower than that before chemotherapy (P =0.048).Levels of TGF-β1 and IL-10[(22.25±6.79), (184.85±49.11) ng/ml]were slightly decreased, which showed no significant difference contrast to pretherapy[(24.37±8.10), (197.16±44.57) ng/ml](P =0.276, P =0.314).In study group, levels of CD+4CDhi25CDlow127Treg cells and TGF-β1, IL-10[(4.36±1.19) %,(20.16±4.73), (165.42±39.57) ng/ml]were obviously lower than that before chemotherapy[(5.78±1.50) %,(23.81±5.15), (198.35±43.68) ng/ml](P=0.000, P=0.003, P =0.006).Conclusion The CD+4CDhi25CDlow127 Treg cells in the peripheral blood of advanced NSCLC patients is significantly increased. Aidi injection combined with chemotherapy can obviously reduce the level of CD+4CDhi25CDlow127 Treg cells in patients with advanced NSCLC, and the combined therapy could improve the immune function of patients.

OBJECTIVETo investigate the impact of age and gender on multiple organ dysfunction syndrome (MODS) after multiple injuries and the possible mechanism.

METHODSThe clinical data of 78 patients with multiple injuries admitted in the ICU in our hospital from Jan. 2004 to Jan. 2007 were reviewed for age, gender, ISS, incidence of MODS, sepsis and mortality, days in ICU, and duration of ventilation. The plasma levels of TNF-alpha and IL-6 were examined by ELISA in the first two weeks after the injury, and the results were analyzed in relation to the demographic and clinical data.

RESULTSWith similar ISS scores, the male patients were more likely to develop into MODS than the female patients (P<0.05), and age produced a marginal effect on the incidence of MODS (P=0.06). Compared with patients without MODS, those with MODS had a significantly different pattern of plasma TNF-alpha and IL-6 alterations. In the initial week following the injury, the male patients showed significantly higher plasma IL-6 levels than the female patients with the same ISS scores (P=0.04), but the level alteration in the initial two weeks showed no significant difference between the male and female patients (P=0.14).

CONCLUSIONGender plays an important role in the occurrence of MODS following multiple injuries, but the effect of age on MODS seems only marginal and need to be further investigated.

Adolescent ; Adult ; Age Factors ; Aged ; China ; epidemiology ; Cytokines ; blood ; Female ; Humans ; Interleukin-6 ; blood ; Male ; Middle Aged ; Multiple Organ Failure ; blood ; epidemiology ; etiology ; Multiple Trauma ; blood ; complications ; Sex Factors ; Tumor Necrosis Factor-alpha ; blood ; Young Adult

ObjectiveTo investigate the temporal features of renal injury in rats with severe heat stroke (SHS) and their relationship with inflammatory response.Methods Fifty-six male Wistar rats were randomly divided into normal control group and SHS for 0, 2, 6, 24, 48, 72 hours group (SHS-0, 2, 6, 24, 48, 72 h groups), with 8 rats in each group. Rats were placed in an artificial climate chamber [temperature (39.5±0.2)℃, humidity (60±5)%] to induce SHS model, and the criterion for successful model reproduction was the onset of lowering peak systolic blood pressure (SBP). Then the rats were transferred to room temperature (23.0±0.2)℃ after successful reproduction of the model. The rats of normal control group were kept in room temperature of (23.0±0.2)℃. Heart blood and renal tissue samples were harvested, and the levels of serum creatinine (SCr) and blood urea nitrogen (BUN) were determined by automatic biochemistry analyzer. The levels of myeloperoxidase (MPO), tumor necrosis factor-α(TNF-α) and interleukin-6 (IL-6) in renal tissue specimens were determined by enzyme linked immunosorbent assay (ELISA). The changes in histopathology in kidney were observed with light microscopy, and Paller scores were used to assess the degree of renal injury.Results Compared with normal control group, the levels of SCr and BUN in serum, and MPO, TNF-α and IL-6 in the renal tissue homogenate were significantly increased in SHS-6 h group [SCr (μmol/L): 174.0±27.0 vs.68.0±11.3, BUN (mmol/L): 12.6±2.3 vs. 4.3±1.2, MPO: (203.0±38.0)% vs. (100.0±1.4)%, TNF-α: (121.0±16.0)% vs. (100.0±1.4)%, IL-6: (118.0±19.0)% vs. (100.0±1.3)%, allP< 0.05], and they peaked at 24 hours [SCr (μmol/L): 489.0±96.0 vs. 68.0±11.3, BUN (mmol/L): 19.3±5.7 vs. 4.3±1.2, MPO: (511.0±41.0)% vs. (100.0± 1.4)%, TNF-α: (399.0±47.0)% vs. (100.0±1.4)%, IL-6: (473.0±56.0)% vs. (100.0±1.3)%, allP< 0.01], then declined to the normal levels at 72 hours. Under light microscopy, tissue edema and necrosis of renal tubules were found, and leukocyte infiltration was found to be most profuse at 24 hours, then they returned to normal levels at 72 hours. Paller scores in SHS-6 h group were significantly higher than those of the normal control group (75.45±9.70 vs. 14.23±3.26,P< 0.01), and it peaked at 24 hours (186.00±14.25 vs. 14.23±3.26,P< 0.01), followed by a gradual lowering, back to normal level at 72 hours.ConclusionThe results suggest that progressive renal damage occurred in the rats with SHS within 24 hours, and it was accompanied with elevated levels of MPO, TNF-α and IL-6 in the kidney homogenate, suggesting that inhibition of neutrophil activation and the release of IL-6, TNF-α may protect the SHS associated renal injury.

To investigate cytotoxic secondary metabolites of Micrococcus sp. R21, an actinomycete isolated from a deep-sea sediment (-6 310 m; 142 degrees 19. 9' E, 10 degrees 54. 6' N) of the Western Pacific Ocean, column chromatography was introduced over silica gel, ODS, and Sephadex LH-20. As a result, eight compounds were obtained. By mainly detailed analysis of the NMR data, their structures were elucidated as cyclo(4-hydroxy-L-Pro-L-leu) (1), cyclo(L-Pro-L-Gly) (2), cyclo( L-Pro-L-Ala) (3), cyclo( D-Pro-L-Leu) (4), N-β-acetyltryptamine (5), 2-hydroxybenzoic acid (6), and phenylacetic acid (7). Compound 1 exhibited weak cytotoxic activity against RAW264. 7 cells with IC50 value of 9.1 μmol x L(-1).
Animals ; Biological Factors ; chemistry ; isolation & purification ; metabolism ; pharmacology ; Cell Survival ; drug effects ; Macrophages ; cytology ; drug effects ; Magnetic Resonance Spectroscopy ; Mass Spectrometry ; Mice ; Micrococcus ; chemistry ; genetics ; isolation & purification ; metabolism ; Molecular Structure ; Phylogeny ; RAW 264.7 Cells ; Seawater ; microbiology ; Secondary Metabolism