OBJECTIVETo investigate the effect of resveratrol (RSV) on angiotensin II (Ang II) induced cardiomyocytes hypertrophy and FoxO1/MnSOD signaling pathway.

METHODSThe cardiomyocytes isolated from neonatal Wistar rats were cultured with pancreatin and preplating technique and divided into five groups: control (CON), Ang II (1 x 10(-6) mol/L, Ang II), Ang II + RSV (10 µmol/L), Ang II + RSV (25 µmol/L), and Ang II + RSV (50 µmol/L). 3H-Leucine incorporation method were used to determine the cardiomyocyte protein synthesis rate. Cell size was measured by phase contrast microscope. The gene expression of A type natriuretic factor (ANF) was detected by real-time PCR. Western blot was used to measure the expression of MnSOD, FoxO1 and acety-FoxO1. Immunoprecipitation was used to detect the interaction between Sirt1 and FoxO1.

RESULTSCardiomyocyte protein synthesis rate in Ang II group was significantly higher in Ang II group than in the control group ((1,971 ± 175) cpm vs. (1,216 ± 136) cpm, P < 0.05), which could be significantly attenuated by RSV (10 µmol/L, 25 µmol/L, 50 µmol/L, all P < 0.05), especially in Ang II + RSV (25 µmol/L) group( (1,374 ± 143) cpm). Cardiomyocytes size in Ang II group was significantly larger than control group ((29.3 ± 3.2) µm vs. (19.4 ± 1.8) µm, P < 0.05), which could be significantly reduced by cotreatment with RSV (10 µmol/L, 25 µmol/L, 50 µmol/L, all P < 0.05), especially in Ang I + RSV (25 µmol/L) group ((20.8 ± 2.1) µm). Ang II also significantly upregulated ANP mRNA expression of cardiomyocytes (4.4 ± 0.4 vs. 1.0 ± 0.1 in control group, P < 0.05), which could be significantly inhibited by cotreatment with RSV, especially in Ang II + RSV (25 µmol/L) group (2.2 ± 0.2). Ang II significantly decreased MnSOD expression of cardiomyocytes compared with control group (P < 0.05), which was reversed by RSV (25 µmol/L). The binding level of Sirt1 and FoxO1 was significantly lower (1.00 ± 0.11 vs. 1.63 ± 0.16, P < 0.05), and the expression of acetylation of FoxO1 was significantly higher in Ang II group than in control group (1.48 ± 0.16 vs. 1.00 ± 0.13, P < 0.05), which was significantly reversed by cotreatment with RSV (25 µmol/L).

CONCLUSIONSResveratrol treatment can inhibit Ang II induced cardiomyocyte hypertrophy. This protective effect is associated with reduced FoxO1 acetylation and activation of Sirt1, suggesting that Sirt1 may serve as a potential therapeutic target of cardiomyocyte hypertrophy.

Angiotensin II ; Animals ; Animals, Newborn ; Forkhead Transcription Factors ; Hypertrophy ; Myocytes, Cardiac ; Nerve Tissue Proteins ; Rats ; Rats, Wistar ; Signal Transduction ; Stilbenes ; Superoxide Dismutase

AIM: To study the serum level of interferon ( IFN)-?, IFN-?, nitric oxide (NO) and nitric oxied synthase (iNOS) in severe ac ute respiratory syndrome (SARS) patients and to expl ore its significance. METHODS: IFN-?, IFN-?, NO and iNOS were determined in SARS patients during the early, recovery and follow-up stage, the health doctors and nurses and health people, respectively. RESULTS : The serum level of IFN-? during the early stage was higher than that of other groups (P

Objective To investigate the effect of Sirt1 gene knockout on chronic kidney disease induced by 5/6 nephrectomy in mice and vascular endothelial growth factor (VEGF)/fetal liver kinase-1 (Flk-1) signaling pathway.Methods Twenty four male Sirt1 +/+ and Sirt1 +/-mice wererandomly divided into four groups:Sirt1+/+ mice with sham-operation (WT-Sham,n=6),Sirt1+/-mice with sham-operation (KO-Sham,n=6),Sirt1 +/+ mice with 5/6 nephrectomy (WT-Nx,n=6) and Sirt1 +/-mice with 5/6 nephrectomy (KO-Nx,n=6).Proteinuria was determined by urine collection from 8:00 to 8:00 the next day at 20 weeks.Serum creatinine (Scr),urea nitrogen (BUN) and the renal pathological changes were measured after 20 weeks.Expressions of Sirt1,collagen Ⅰ and transforming growth factor β(TGF-β) were used to analyze the changes of renal fibrosis by immunohistochemistry staining.Real-time PCR and Western blotting were used to measure the mRNA and protein expressions of Sirt1,fibronectin,collagen Ⅰ,VEGF and Flk-1 in kidney.Results Sirt1 expressed in glomernlar endothelial cells,podocytes,mesangial cells and renal tubular epithelial cells in Sirt1 +/+ mice,while Sirt1 expression intensity was significantly reduced in Sirt1 +/-mice.Compared with the WT-Sham group,WT-Nx group had increased proteinuria,BUN,Scr,glomernlar sclerosis index and tubulointerstitial fibrosis index at 12 weeks after operation (all P ＜ 0.01),and KO-Nx group had exacerbated the above up-regulations (all P ＜ 0.01).Compared with those in WT-Sham group,the expressions of fibronectin,collagen Ⅰ and TGF-β were up-regulated in WT-Nx group (all P ＜ 0.01),and were significantly augmented in KO-Nx group (all P ＜ 0.01).Compared with those in WT-Sham group,renal mRNA and protein expressions of VEGF and Flk-1 were decreased in WT-Nx group,and KO-Nx group aggravated their down-regulation (all P ＜ 0.01).Conclusions Sirt1 gene knockout can increase proteinuria and Scr,and aggravate renal pathology and renal fibrosis in 5/6 nephrectomized mice,which is associated with the inhibition of VEGF/Flk-1 signaling pathway.It is suggested that Sirt1 may be a potential therapeutic target of chronic kidney disease.

Objective To express adenosine deaminase protein by molecular cloning technique.MethodsTotal RNA was extracted from human leukocytes and the cDNA was obtained by reverse transcription.Whereupon the cDNA was used as a template to amplify adenosine deaminase gene by polymerase chain reaction (PCR) and then integrated it into three prokaryotic plasmids pET-28 b, pET-32 a (+) and pHSIE.The plasmid with the correct sequencing was transformed into E.coli BL21 (DE3) by CaCl2 method for the protein expression.The expression activity of these fusion proteins were detected by Western-blot and SDS-PAGE, with the optimized expression conditions.Results Complete fusion of target gene and three prokaryotic plasmids was observed through sequencing.The expressed and accurate ADA protein was identified by Westernblot and SDS-PAGE.The optimal expression conditions were observed:the protein expression would be induced with 0.4 mmol/L IPTG and incubated at 16℃for 24 hours.Conclusion The prokaryotic vectors of adenosine deaminase (BL21+pET-28 b+ADA, BL21+pET-32 a+ADA, BL21+pHSIE+ADA) were successfully constructed and efficiently expressed.

Objective To investigate the neuroprotective effect and its mechanism of p38 mitogenactivated protein kinase inhibitor after subarachnoid hemorrhage (SAH).Methods Twenty-seven SPF-grade adult male SD rats were selected to induce a SAH model using the prechiasmal pool blood injection.Three dead rats were excluded.Twenty-four rats were randomly divided into four groups:sham operation group,SAH group,dimethyl sulfoxide (DMSO) group,and DMSO +p38 inhibitor group (n =6 in each group).Western blot was used to detect the expression levels of p38,phosphorylation p38,Parkinson's disease protein 7 (DJ-1),autophagy associated gene 5 (Atg5),autophagy adaptor protein p62,microtubule-associated protein Ⅰ Light Chain 3 (LC3-Ⅰ),microtubule-associated protein Ⅱ light chain 3 Ⅱ (LC3-Ⅱ),and the Garcia neurological function score was used to judge the nerve injury.PC12 cell oxygenated hemoglobin was used to induce an in vitro SAH model.They were completely randomly divided into four groups:sham operative group,SAH group,DMSO group,and DMSO + p38 inhibitor group.Fluorescent probe JC-1 was used to observe the changes of mitochondrial membrane potential.Results (1) There were significant differences in the expression of p38,phosphorylation-p38 and DJ-1 in rat brain tissue among the 4 groups (F values were 94.959,150.293 and 698.476,respectively,all P ＜ 0.01).There were significant differences in mitochondrial membrane potential in PC12 cells among the 4 groups (F value was 24.989,P ＜ 0.01).There were significant differences in the expression levels of autophagy related protein LC3-Ⅱ/LC3-Ⅰ ratio,Atg5 and p62 in rat brain tissue among the 4 groups (F values were 235.319,110.490 and 36.311,respectively,all P ＜ 0.01).There was significant difference in nerve function score among the 4 groups (F value was 25.550,P ＜ 0.01).(2) Compared with the sham operative group,the expression levels of p38,phosphorylation-p38 and DJ-1 were upregulated significantly after SAH (from 0.43 ±0.06,0.41 ±0.02 and 0.07 ±0.01 to 0.61 ± 0.08,0.79 ± 0.07 and 0.17 ± 0.03,respectively,all P ＜ 0.01),mitochondria membrane potential depolarization (from 8.29 ±0.28 to 9.23 ±0.42,P ＜0.01);upregulation of Atg5 expression and increase of LC3-Ⅱ/LC3-Ⅰratio (from 0.23 ± 0.04 and 0.25 ± 0.04 to 0.47 ± 0.04 and 0.46 ± 0.04,respectively,all P ＜ 0.01),down regulation of p62 expression (from 1.09 ± 0.14 to 0.54 ± 0.10,P ＜ 0.01);neurological score was decreased (from 17.5 ± 0.6 to 11.3 ± 2.7,P ＜ 0.01);p38 inhibitor was significantly down regulated the expression of phosphorylation-p38 after SAH (from 0.79 ± 0.07 to 0.47 ± 0.04,P ＜ 0.01),the expression of DJ-1 was up-regulated (from 0.17 ± 0.03 to 1.02 ± 0.06,P ＜ 0.01),mitochondrial membrane potential recovery (from 9.23 ±0.42 to 8.47 ±0.36,P ＜0.01),cell autophagy related protein LC3-Ⅱ/LC3-Ⅰ ratio and Atg5 were upregulated(from 0.46 ±0.04 and 0.47 ±0.04 to 0.77 ± 0.06 and 0.95 ± 0.12,all P ＜ 0.01),p62 expression returned to the levels of SAH group (from 0.57 ± 0.09,to 0.54 ± 0.10,P =0.650),and the neurological score was significantly improved (from 11.3 ± 2.7 to 15.5 ± 1.0,P ＜0.01).Conclusions After SAH,the p38 inhibitor downregulates the activity of2 phosphorylation p38.It may inhibit abnormal autophagy and maintain mitochondrial function by up-regulating the expression of DJ-1 protein,and then play a neuroprotective function.

Objective:To compare four reconstruction algorithms of 18F-fluorodeoxyglucose (FDG) PET/CT on standardized uptake value (SUV) of pulmonary nodules. Methods:A total of 46 patients (27 males, 19 females; median age: 66 (range: 44-82) years) with solid pulmonary nodules from February 2018 to July 2019 in the First Hospital of Shanxi Medical University who performed 18F-FDG PET/CT imaging were enrolled. All PET/CT images were retrospectively reconstructed by using four algorithms reconstructions including ordered subset expectation maximization (OSEM), OSEM+ time of flight (TOF), OSEM+ TOF+ point spread function (PSF) and block sequential regularized expectation maximization (BSREM) (G1-G4). Nodule and background parameters were analyzed semi-quantitatively and visually. The maximum of SUV(SUV max), mean of SUV(SUV mean) and peak of SUV (SUV peak) were collected by the region of interest (ROI). Nodules were divided into small nodule group (diameter ≤10 mm) and large nodule group (10 mm < diameter ≤30 mm). Kruskal-Wallis rank sum test and Bonferroni method were performed to compare the differences of SUVs between G1-G4, and Spearman correlation analysis was used to analyze the correlation between the change rate of SUV (%ΔSUV) and the diameter of nodules. The receiver operating characteristic (ROC) curve analysis was used to analyze the diagnostic efficacy of SUV for the differential diagnosis of pulmonary nodules and to get the optimal threshold. Results:There were 114 pulmonary nodules (large nodules, n=55; small nodules, n=59). In visual analysis, the visual detection rates of small nodules in G4 were 55.93%(33/59), 44.07%(26/59), 20.34%(12/59) higher than those in G1-G3. Of 114 pulmonary nodules in 46 patients, there were differences in SUV max and SUV mean between G1-G4 (median SUV max : 2.65-5.29, median SUV mean: 2.05-2.99; H values: 20.628 and 17.749, respectively, both P<0.001), G4 had significant increases compared to G1 in SUV max (median 5.29 and 2.65, P<0.001) and SUV mean (median 2.99 and 2.05, P<0.001). The %ΔSUV max (median: 4.45%-52.96%) and %ΔSUV mean (median: 1.69%-47.56%) were negatively correlated with the diameter of nodules (9.75(6.20, 16.58) mm; r s values: -0.371 to -0.354, -0.371 to -0.320, all P<0.001). In 59 small nodules, G1 significantly increased the SUV max of G4 (median 4.05 and 2.14, H=18.327, P<0.001), while G4 significantly increased the SUV mean of G1 and G3 (median 2.31, 1.26 and 1.53, H=16.808, P<0.05). There was no significant difference in SUVs between G1-G4 in 55 large nodules ( H values: 0.812-7.290, all P>0.05). The optimal threshold values of SUV max in G1-G4 were 4.335, 5.185, 5.410, 5.745 and the area of under curves (AUCs) were 0.747, 0.699, 0.756, 0.778 respectively. The AUC of SUV mean and SUV peak also showed a similar trend. Conclusion:Among the four reconstruction algorithms, BRERM can not only enhance the image quality, but also significantly improve the SUV max and SUV mean of lung nodules diameter below 10 mm, and thus its diagnostic threshold of SUV should be appropriately increased.