Crocetin, a naturally occurring carotenoid, possesses antioxidant and antiatherosclerotic properties, of which the underlying mechanism remains unclear. In the present study, we examined the effects of crocetin (0.1, 1, 10 μmol·L(-1)) on angiotensin II (Ang II, 0.1 μmol·L(-1)) induced expression of vascular cell adhesion molecule-1 (VCAM-1) in human umbilical vein endothelial cells (HUVECs) and monocyte-endothelial cell adhesion. The effects of crocetin on the activation of nuclear factor kappa B (NF-κB) and intracellular reactive oxygen species (ROS) were also observed. The results demonstrated that crocetin notably suppressed Ang II induced NF-κB activation (P<0.01) and VCAM-1 expression (P<0.05, P<0.01) in HUVECs, accompanied by a markedly reduced monocyte-endothelial cell adhesion (P<0.05, P<0.01). In addition, preincubation with crocetin resulted in a significant enhancement of cellular antioxidant capacity (P<0.05, P<0.01), while Ang II induced intracellular ROS decreased markedly (P<0.05, P<0.01). These results indicated that crocetin was capable of suppressing Ang II induced VCAM-1 expression and monocyte-endothelial cell adhesion by suppression of NF-κB activation, which might be derived from the enhancement of antioxidant capacity and subsequent reduction of intracellular ROS.
Angiotensin II ; metabolism ; Antioxidants ; pharmacology ; Carotenoids ; pharmacology ; Cell Adhesion ; drug effects ; Human Umbilical Vein Endothelial Cells ; cytology ; drug effects ; metabolism ; Humans ; Monocytes ; cytology ; NF-kappa B ; metabolism ; Reactive Oxygen Species ; metabolism ; Vascular Cell Adhesion Molecule-1 ; metabolism

Biomarkers, Tumor ; metabolism ; Carcinoma, Non-Small-Cell Lung ; drug therapy ; metabolism ; pathology ; Humans ; Lung Neoplasms ; drug therapy ; metabolism ; pathology ; Oncogene Proteins, Fusion ; chemistry ; metabolism ; Protein Kinase Inhibitors ; therapeutic use ; Pyrazoles ; therapeutic use ; Pyridines ; therapeutic use ; Pyrimidines ; therapeutic use ; Smoking

Aluminum ; toxicity ; Animals ; Animals, Newborn ; Calcium ; metabolism ; Cerebral Cortex ; drug effects ; metabolism ; Dose-Response Relationship, Drug ; Female ; Male ; Rats ; Rats, Wistar

OBJECTIVETo observe the effect of sesamin (Ses) on pulmonary vascular remodeling in rats with monocrotaline ( MCT)-induced pulmonary hypertension (PH).

METHODTotally 48 male Sprague-Dawley (SD) rats were fed adaptively for one week and then divided into the normal control group, the MCT group, the MCT +Ses (50 mg x kg(-1)) group and the MCT + Ses (100 mg x kg(-1)) group, with 12 rats in each group. The PH rat model was induced through the subcutaneous injection with MCT(60 mg x kg(-1)). After the administration for four weeks, efforts were made to measure the right ventricular systolic pressure( RVSP) and mean pulmonary artery pressure (mPAP) through right jugular vein catheterization, and isolate right ventricle( RV) and left ventricle( LV) +septum (S) and measure their length to calculate RV/ ( LV + S) and ratio of RV to tibial length. Pathologic changes in arterioles were observed by HE staining. Masson's trichrome stain was used to demonstrate changes in collagen deposition of arterioles. The alpha-smooth muscle actin (alpha-SMA) expression in pulmonary arteries was measured by immunohistochemisty. The total antioxidative capacity (T-AOC) and malondialdehyde (MDA) content in pulmonary arteries were determined by the colorimetric method. The protein expressions of collagen I, NOX2 and NOX4 were analyzed by Real-time PCR and Western blot.

RESULTAfter the administration for 4 weeks, Ses could attenuate RVSP and mPAP induced by MCT, RV/ (LV + S) and ratio of RV to Tibial length, alpha-SMA and collagen I expressions and remodeling of pulmonary vessels and right ventricle. Meanwhile, Ses could obviously inhibit the expressions of NOX2, NOX4 and MDA content and increase T-AOC.

CONCLUSIONSesamin could ameliorate pulmonary vascular remodeling induced by monocrotaline in PH rats. Its mechanism may be related to expressions of NOX2 and NOX4 expression and reduction in oxidative stress injury.

Animals ; Dioxoles ; administration & dosage ; Disease Models, Animal ; Drugs, Chinese Herbal ; administration & dosage ; Humans ; Hypertension, Pulmonary ; drug therapy ; enzymology ; genetics ; physiopathology ; Lignans ; administration & dosage ; Lung ; blood supply ; enzymology ; metabolism ; Male ; Membrane Glycoproteins ; genetics ; metabolism ; Monocrotaline ; adverse effects ; NADPH Oxidase 2 ; NADPH Oxidase 4 ; NADPH Oxidases ; genetics ; metabolism ; Pulmonary Artery ; drug effects ; metabolism ; physiopathology ; Rats ; Rats, Sprague-Dawley ; Vascular Remodeling ; drug effects

Objective To establish a simple and economic method for isolating rat hepatic lipocytes. Method Perfusion of the rat liver with pronase E and collagenase in succession was performed in cycles, and the hepatic lipocytes were isolated and purified by digesting the rat liver with DNase in the presence of the above 3 enzymes, followed by 11% Necodenz density gradient centrifugation. The resulted lipocytes were subjected to routine identification procedures and viability assessment. Results The yield of the lipocytes from each rat was 2×107 to 3×107, with viability over 95% and purity above 90%. Conclusion The method adopted in this study is simple, economic and highly productive.

Objective To identify the important risk factors for type 2 Diabetes Mellitus (T2DM) and develop effective strategies to address the problem of T2DM.Our study aimed to evaluate the association between apolipoprotein E (ApoE) genetic polymorphism and type 2 diabetes,and to provide clues for the etiology of T2DM.Methods Based on the criteria of inclusion and exclusion,we extracted,pooled,analyzed and assessed the case-control studies of ApoE polymorphism and T2DM published in PubMed,Web of Science,Medline,WanFang,VIP,and CNKI databases by R soft-ware (version 3.4.3).We used Random-effect models when heterogeneity was present in between-study,and fixed-effect models otherwise.Results We had 59 studies covering 6,872 cases with T2DM and 8,250 controls,and compared the alleles and genotypes of ApoE between cases and controls.When we conducted a comparison between ApoE ε4 and ε3 alleles,we produced a pooled OR of 1.18 (95％ CI:1.09-1.28;P ＜ 0.001).ApoE ε2/ε2 genotype displayed a possible association with T2DM (OR =1.46;95％ CI:1.11-1.93;P =0.007),ε3/ε4 genotype showed a 1.11-fold risk (OR =1.11;95％ CI:1.01-1.22;P =0.039) and ε4/ε4 genotype had a 1.71-fold risk of developing T2DM (OR =1.71;95％ CI:1.33-2.19;P ＜ 0.001) when they were compared with ε3/ε3 genotype.Conclusions There is an association between ApoE polymorphism and T2DM:allele ε4 and genotypes (ε2/ε2,ε3/ε4,and ε4/ε4) are associated with the increased risk for the development of T2DM,and they may be risk factors for T2DM.

Objective To establish a simple and economic method for isolating rat hepatic lipocytes. Method Perfusion of the rat liver with pronase E and collagenase in succession was performed in cycles, and the hepatic lipocytes were isolated and purified by digesting the rat liver with DNase in the presence of the above 3 enzymes, followed by 11% Necodenz density gradient centrifugation. The resulted lipocytes were subjected to routine identification procedures and viability assessment. Results The yield of the lipocytes from each rat was 2×107 to 3×107, with viability over 95% and purity above 90%. Conclusion The method adopted in this study is simple, economic and highly productive.

Objective To understand the characteristics of cervix diseases in female medical staff and provide basis for effective prevention and control.Methods The results of cervical cancer screening in 1 471 female medical staff in our hospitals in 2008 were compared with the 1 706 ones in 2010.Results Morbidity rates of various cervix diseases were 37.9％ (558/1 471) in 2008 and 50.6％ (864/1 706) in 2010,respectively.Most of them were diagnosed with chronic cervicitis (36.8％ in 2008 and 47.1％ in 2010).The difference of positive rate of screening test (TCT results ≥ASC-US) was significant between those in 2008( 1.1％,16/1 471 )and those in 2010(3.7％,63/1 706) ( x2 =12.65,P ＜ 0.01 ).The results also showed that the incidences of various cervix diseases were related to age,and that high rates of cervicitis and cervical cancer were observed in the middle-aged (35 -50 years old).Conclusions Positive rates of cervical cancer screening test tend to rise,and the health condition of the middle-aged women (35 -50 years old) should be paid particular attention.

OBJECTIVETo evaluate the role of RNA interference (RNAi) in silencing the enhanced green fluorescent protein (eGFP) expression in 293T and Mel cells.

METHODSNested-PCR was used to amplify H1 promoter from human 293T cells for driving RNAi synthesis. RNAi vectors (TR1) for silencing the eGFP expression was constructed. The eGFP vector and RNAi vector (TR1) were then co-transfected into the 293T and Mel cells, in which the silencing effect on eGFP expression was investigated by fluorescence microscopy, reverse transcription-PCR(RT-PCR), fluorescence-assited cell sorting(FACS) analysis and real-time RT-PCR.

RESULTSRNAi could effectively reduce more than 50 percent of eGFP expression in 293T cells as well as in Mel cells.

CONCLUSIONThe RNAi vector constructed in this way paper can effectively inhibit eGFP expression in cells.

Cell Line ; Flow Cytometry ; Genetic Vectors ; genetics ; Green Fluorescent Proteins ; genetics ; Humans ; RNA Interference ; Reverse Transcriptase Polymerase Chain Reaction

OBJECTIVETo investigate the protective effects of rutaecarpine (Rut) on right ventricular remodeling in rats with monocrotaline-induced pulmonary hypertension (PH).

METHODForty-eight SD rats were fed adaptively for 1 week and then were randomly divided into the following 4 groups (n = 12): normal control group, monocrotaline (MCT) treatment group, MCT treatment with Rut (20 mg/kg)group and MCT treatment with Rut (40 mg/kg) group. PH rats were induced by a single injection of monocrotaline (60 mg/kg, sc) and were administered with Rut (20 or 40 mg/kg/d) for 4 weeks. At the end of experiment, the right ventricular systolic pressure (RVSP) and mean pulmonary artery pressure (mPAP) were monitored via the right jugular vein catheterization into the right ventricle. The ratio of right ventricle (RV) to left ventricle (LV) + septum (S) and the ratio of RV to tibial length were calculated. Right ventricular morphological changes were deserved by HE staining. Masson's trichrome staining was used to display collagen deposition. The total antioxidative capacity (T-AOC) and malondialdehyde (MDA) levels in right ventricle were determined according to the manufacturer's instructions. mRNA and protein expression levels of NOX4, collagen I and collagen III were analyzed by immunohistochemisty, real-time PCR and Western blot.

RESULTSThe results showed that Rut treatment for 4 weeks attenuated RVSP, mPAP and right ventricular remodeling index (RV/LV + S and RV/Tibial length) of PH rats induced by monocrotaline. Furthermore, the right ventricular collagen deposition and collagen I and collagen I expression induced by MCT were both significantly suppressed by Rut. The expression levels of NOX4 and MDA were obviously decreased, while the T-AOC was significantly increased in right ventricular from PH rats treated with Rut.

CONCLUSIONThese results suggested that Rut ameliorates the right ventricular remodeling in rats with PH induced by MCT through down-regulating of NOX4 expression and collagen accumulation.

Animals ; Antioxidants ; metabolism ; Heart Ventricles ; metabolism ; Hypertension, Pulmonary ; chemically induced ; drug therapy ; Indole Alkaloids ; pharmacology ; Male ; Malondialdehyde ; metabolism ; Monocrotaline ; adverse effects ; NADPH Oxidase 4 ; NADPH Oxidases ; metabolism ; Quinazolines ; pharmacology ; Rats ; Ventricular Remodeling ; drug effects