Objective To investigate the effect of olmesartan medoxomil on renal oxidative stress in mice with chronic heart failure. Methods C57 mice were divided into sham operation group(SHAM group),chronic heart failure group(CHF group)and olmesartan medoxomil treatment group(OLM group).Experimental CHF model was established by coronary artery ligation,in which OLM group fed with a daily dose of 10 mg/kg.The heart rate,blood pressure,cardiac function,Scr,BUN,and plasma and kidney angiotensin(Ang)Ⅱ were measured.Real-time PCR was used to examine renal gp91phox,p22phox and NOX4 expression.AZAN and DHE staining was used to detect renal pathological change after 12 weeks. Results Compared with SHAM group,left ventricular-end diastolic dimension (LVDd)and left ventricular end-systolic dimension(LVDs)were significantly increased(P<0.05),while fractional shortening(FS)and ejection fraction(EF)were significantly decreased in CHF and OLM groups (P<0.05).Compared with SHAM group,systolic blood pressure,Scr,BUN,and AZAN and DHE staining positive area were significantly increased in CHF group(P<0.05),while above indexes were significantly lower in OLM group as compared to CHF group(P<0.05).Compared with SHAM group,plasma and kidney Ang Ⅱ levels,gp91phox,p22phox and NOX4 expression were increased in CHF group(P<0.05),while above indexes were significantly lower in OLM group as compared to CHF group (P<0.05).Conclusions Chronic heart failure can activate intrarenal NADPH oxidase resulting in renal injury.Olmesartan medoxomil can protect kidney by inhibiting the effect of Ang Ⅱ-induced oxidative stress.

Objective To explore related influencing factors for postoperative delirium in elderly patients, to provide a reference for the prevention and treatment. Methods Fifty-four patients with delirium after surgery were used as observa?tion group, and a total of 150 subjects with no delirium after surgery during the same period were selected as the control group. Data of age, gender, malnutrition, disorders of water and electrolyte metabolism, postoperative mechanical ventilation, postoperative hypoxemia, severe infection and postoperative pain degree, and the combination of basic diseases were com?pared and analysed between two groups. The binary logistic regression analysis was used to analyse the influencing factors of postoperative delirium. The outcome and prognosis were observed and analyzed in observation group. Results The average age was significantly higher in observation group than that of control group (P<0.05). The percentages of postoperative hy?poxemia and severe infection were significantly higher in observation group than those of control group ( P<0.05). Patients with higher age, postoperative hypoxemia and severe infection were risk factors for postoperative delirium. In observation group,1 case died of lung infection, 1 case died of multiple organ failure, the remaining 52 patients were improved and dis?charged from hospital after three months. Conclusion For patients with higher age, postoperative hypoxemia and severe in?fection are the risk factors for occurrence of postoperative delirium. More attention should be paid to clinical preoperative and postoperative periods.

Objective To investigate the role of the clock gene promoter methylation in aging. Methods C57BL mice of 4- (young, n=9) and 20- (old, n=10) month-old were determined the promoter methylation level of clock genes (Per1/2, Bmal1/2, Cry1/2, Clock, Npas2) in the stomach, spleen, vascular, kidney and striatum with methylation-specific polymerase chain reaction (MSP). Results The incidence of promoter methylation of Cry1, Bmal2 and Npas2 in spleen increased in old mice (P<0.05), while the promoter methylation of Per1 in stomach decreased (P<0.05), and the promoter methylation of Bmal1 in vascular increased (P<0.05). Conclusion Promoter methylation of some clock genes is involved in process of aging in a tissue-specific way.

Objective To investigate the effects of type 2 diabetes on learning and memory of APP/PS1/Tau triple transgenic (3 × Tg) mice of Alzheimer’s disease, and the protective mechanism of liraglutide (LIR) thereof. Methods One month old C57BL/6 mice were set to be control group (WT). One month old 3×Tg mice were divided into control group (Tg), liraglutide group (Tg+LIR), type 2 diabetes group (Tg+T2DM) and liraglutide treatment group (Tg+T2DM+LIR). The model of T2DM was established by feeding the high fat and sugar fodder, and then injecting streptozotocin (STZ) in mice, making sure the fasting blood glucose was more than 7 mmol/L. Then the subcutaneous injection of LIR was administered for 2 months. The values of body weight and fasting blood glucose were detected at age of 5-month. Morris water maze was applied to evaluate the spatial learning and memory ability. Western blotting assay was used to measure the levels of phosphorylated Tau, neurofilament (NFs) and insulin receptor substrates. ELISA was used to detect the human Aβ42 to evaluate the effect of LIR on-amyloid. Results LIR can reduce body weight and blood glucose, can alleviate spatial learning and memory damaging caused by T2DM, and also can improve phosphorylated Tau levels, NFs and insulin receptor substrates caused by T2DM, and finally can reduce the deposition ofβ-amyloid of 3 × Tg mice. Conclusion T2DM can aggravate symptoms of AD in 3×Tg mice, and LIR has a protective effect on it.

Objective To investigate the effects of penehyclidine hydrochloride (PHCD) pretreatment on NF-κB mRNA expression and SOD activity in lung tissues in rats with acute lung injury (ALI) induced by LPS.Methods Thirty-two male SD rats, 2 months old, weighing 230-280 g, were randomly divided into 4 groups (n=8 each): control group (group C), ALI group, low dose PHCD group (group LP) and high dose PHCD group (group HP). ALI was induced by intravenous LPS 5 mg/kg via tail vein. Group LP and HP received intraperitoneal PHCD 0. 3 and 1 mg/kg respectively 30 min before LPS administration. The rats were killed at 6 h after LPS administration. The lungs were removed immediately for determination of W/D lung weight ratio, lung water content, NF-κB mRNA expression, TNF-α and MDA content, and SOD activity and microscopic examination. Results NF-κB mRNA expression, TNF-α and MDA content, W/D lung weight ratio and lung water content were significantly higher, while SOD activity was significantly lower in group ALI, LP and HP than in group C (P ＜ 0.05). NF-κB mRNA expression, TNF-α and MDA content, W/D lung weight ratio and lung water content were significantly lower, while SOD activity was significantly higher in group LP and HP than in group ALI and HP than in group LP (P ＜ 0.05). The LPS-induced changes were mitigated by pretreatment with low and high doses of PHCD in group LP and HP.Conclusion Pretreatment with PHCD attenuates LPS-induced ALI by downregulating NF-κB mRNA expression, decreasing local inflammatory response and enhancing anti-oxidant activity.

Objective To verify and valuate the performance of small dense low-density lipoprotein cholesterol (sdLDL-C) detection by the direct clearance method and evaluate its preliminary clinical application in acute coronary syndrome (ACS).Methods Case control study:The performance (accuracy,precision,linearity) of sdLDL-C was assessed by direct clearance method.In 143 cases of ACS patients selected from Cardiology Department and Emergency Department of Shangdong Provincial Hospital from April to October in 2016,with 100 cases male,female 43 cases,including acute myocardial infarction (AMI)group of 59 cases,unstable angina pectoris (UAP) group of 84 cases;83 cases of healthy volunteers as a control group selected from health physical examination center of Shandong Provincial Hospital,with 59cases male,female 24 cases.Levels of sdLDL-C,total cholesterol (TCH),triglyceride (TG),low-density lipoprotein cholesterol (LDL-C),high-density lipoprotein cholesterol (HDL-C),apolipoprotein A (ApoA I),apolipoprotein B (ApoB),lipoprotein (a) (Lpa) and high sensitive C-reactive protein (Hs-CRP) were detected separately by automatic biochemical analyzer.Non high density lipoprotein cholesterol (non-HDL-C) equals TCH minus HDL-C.x2 test,t test,one-way ANOVA,Pearson correlation and multiple linear regression analysis were used as statistical methods.Results The within-lot or between-lot variation was 2.85％ and 3.36％.Methodological comparison:regression equation Y =0.984X + 0.018,r2 =0.966,t =-0.191,P =0.850.There was a good linear correlation (Y =1.026X + 0.007,r2 =0.999) between theoretical values and actual detection results in range of 0.15-2.65 mmol/L.SdLDL-C concentrations were positive correlated with TCH,non-HDL,LDL-C,TG,ApoB (r =0.758,0.848,0.839,0.514,0.885,respectively,P ＜0.01),and negative correlated with HDL-C (r =-0.224,P =0.001),but no correlation with APOA I,Lpa and Hs-CRP(r =-0.021,0.050,0.003,respectively,P ＞ 0.05).Multiple linear regression analysis showed that the factors influencing sdLDL-C level were HDL-C,ApoB,LDL-C and TG.The levels of sdLDL-C,TG in the ACS group were significantly higher than those in the control group (t =3.415,4.660,respectively,P ＜ 0.01),but no difference between the two groups in the levels of TCH,non-HDL-C and LDL-C (t=-1.831,-0.452,-1.398,respectively,P ＞0.05).Comparing AMI group with control group,sdLDL-C,TG and Hs-CRP were significantly higher than the control group (P =0.000,0.000,0.000,respectively),but TCH,LDL-C and non-HDL were similar between the two groups (P =0.800,0.320,0.120,respectively);Comparing UAP group with control group,TG and Hs-CRP were higher than control group (P =0.001,0.047,respectively),TCH and LDL-C were significantly lower than the control group (P =0.003,0.008,respectively),but sdLDL-C had no difference (P =0.305);Comparing AMI group with UAP group,sdLDL-C,TCH,LDL-C and Hs-CRP were significantly higher than UAP group (P =0.000,0.003,0.001,0.000,respectively),and TG were no statistical significance (P =0.473).Conclusions Direct clearance method can meet the requirement of sdLDL-C detection.sdLDL-C level can assess the metabolism of blood lipids and be used as an independent risk factor and predictive index of ACS,superior to LDL-C.

Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) is a promising anti-cancer agent. Epigallocatechin-3-gallate (EGCG) is a polyphenolic constituent of green tea. In this study, inhibitory effect of combined use of EGCG and TRAIL on human melanoma A375 cells was examined and the possible mechanism investigated. The cells were divided into 4 groups: control group, EGCG group (EGCG: 10, 20 mug/mL), TRAIL group (TRAIL: 25 ng/mL) and EGCG+TRAIL group (combined group). The growth inhibition was measured in the A375 cells treated with different concentrations of TRAIL ((25, 50, 75, 100, 125, 150 ng/mL) by MTT assay. The apoptosis was assessed by flow cytometry. The expressions of DR4 and DR5 were detected by flow cytometry and western blotting. The activities of caspase-8 and caspase-3 were determined by colorimetric assay. The results showed that TRAIL could dose-dependently inhibit the growth of A375 cells and the IC(50) of TRAIL was 150 ng/mL. The apoptosis rate was 11.8% in the TRAIL group, 5%-7% in the EGCG group and 48.9%-59.1% in the combined group. Significant difference was found in the apoptosis rate between the combined group and the EGCG or TRAIL group (P<0.05 for each). The expression of DR4 instead of DR5 was significantly increased in the EGCG group. The activity of caspase-3 rather than caspase-8 was substantially enhanced in the EGCG group. These results suggest that EGCG is useful for the TRAIL-based treatment for melanoma.

Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) is a promising anti-cancer agent. However, emergence of drug resistance limits its potential use. Plumbagin is a natural quinonoid compound isolated from plant. In this study, induced apoptosis effect of the combined treatment with plumbagin and TRAIL on human melanoma A375 cell line was examined and possible mechanism was investigated. The cells were divided into four groups: control group, plumbagin group (plumbagin, 5 or 10 mumol/L), TRAIL group (TRAIL, 30 ng/mL) and plumbagin+TRAIL group (combined treatment group). The apoptosis, and the expression of DR4 and DR5 were detected by flow cytometry. The activities of caspase-8 and caspase-3 were determined by colorimetric assay. The results showed that the apoptosis rate was 8.3% in TRAIL group, 10.35%-16.94% in plumbagin group and 52.39%-65.39% in combined treatment group, respectively, with the difference being significant between combined treatment group and plumbagin or TRAIL group (P<0.05 for each). Moreover, plumbagin alone could markedly up-regulate DR5 mRNA and protein expression, and slightly increase DR4 mRNA and protein expression. Treatment of human melanoma A375 cells with plumbagin resulted in the activation of Caspase-3, but not Caspase-8. These results suggest that plumbagin might be useful for TRAIL-based treatment for melanoma.

Objective To explore the potential mechanisms of non-small cell lung carcinoma cells to rsTRAIL protein-induced apoptosis by phosphatidylinositol 3′-kinase (PI3K/Akt)inhibitor LY294002,and to provide new ways to increase killing activities of rsTRAIL protein for non-small cell lung cancer.Methods The A549 cells at logarithmic growth phase were selected and randomly divided into rsTRAIL group and LY294002+rsTRAIL group. The inhibitory rate of growth of the A549 cells was tested by MTT assay.The cell cycle and apoptotic rate were detected by flow cytometry analysis. The expression levels of Ser473 phosphorylated form of Akt (p-Akt),c-FLIPL protein and Bcl-2 protein in the A549 cells in two groups were analyzed by Western blotting method. Results The inhibitory rate of growth of the A549 cells in LY294002+rsTRAIL group (74.6 %± 2.63%)was higher than that in rsTRAIL group (5.61% ± 0.32%) (P< 0.05 ). Compared with rsTRAIL group, the percentage of the cells at G0/G1 phase in LY294002+rsTRAIL group was increased(P<0.05)and the percentage of the cells at S phase was decreased(P<0.05).The apoptotic rate of the A549 cells in LY294002+rsTRAIL group (61.5%±3.02%)was higher than that in rsTRAIL group (3.21%±0.96%)(P<0.05). The Western blotting results showed that the expression levels of p-Akt, c-FLIPL and Bcl-2 proteins in the A549 cells in LY294002+rsTRAIL group were decreased (P<0.05 )and the ratio of Bax/Bcl-2 was increased (P<0.05 ) compared with rsTRAIL group.Conclusion LY294002 can increase the killing activity of rsTRAIL protein in A549 cells by inhibiting the activity of PI3K.

Objective:To explore the proportions of Th17 cells in the peripheral blood and levels of IL-17,IL-23 in the serum of patients with Graves′disease ( GD ) and their clinical significance.Methods: We studied 29 patients with GD ( GD group ) , and reevaluated the GD group after therapy ( euthyroid GD group ).29 gender-and age-matched volunteers were selected as the normal control ( NC group).The proportions of Th17 cells were investigated by flow cytometry.The levels of IL-23,IL-17 in the serums were measured by ELISA.The levels of FT3,FT4,TSH were determined by ECLIA and the levels of TrAb were tested by RRA.Results:There were no significant difference among 3 study groups in sex and age match ( F=0.0075 , P>0.05;χ2=0.4213 , P>0.05 ).The proportions of Th17 cells and the levels of IL-17 , IL-23 were increased in the GD and euthyroid GD patients compared with the control group (respectively,P<0.05).The proportions of Th17 cells and the concentrations of IL-23 in euthyroid GD group were significantly lower than those of GD group ( respectively , P<0.05 ) , but there were no significance in the concentrations of IL-17 between euthyroid GD group and GD group(P>0.05).Correlation analysis revealed that the proportions of Th17 cells ,and the levels of IL-17,IL-23 were positively correlated with the levels of FT3,FT4,TrAb(r=0.588 2,0.337 2,0.371 0;0.549 6,0.287 5,0.342 7;0.361 0,0.420 8, 0.330 8;P<0.05 ,for all parameters ) ,and were negatively correlated with the levels of TSH ( r=-0.319 7 ,-0.472 8 ,-0.428 2;P<0.05,for all parameters).Conclusion:Th17 cells and their related cytokines IL-17,IL-23 are highly expressed in the serum of patients with GD.Th17 cells and their relative cytokines have certain relevance with 4 thyroid function parameters of the patients with GD , which can be used as biological markers for GD.