Animals ; Benchmarking ; methods ; statistics & numerical data ; Dose-Response Relationship, Drug ; Humans ; Models, Statistical ; No-Observed-Adverse-Effect Level ; Risk Assessment ; methods ; statistics & numerical data ; Toxicology ; methods

BACKGROUNDProbucol is known to reduce the development of atherosclerotic lesions, but its impact on vascular remodeling associated with de novo atherosclerosis is incompletely understood. We therefore examined the effect of probucol on vascular remodeling in a rabbit model of established atherosclerosis.

METHODSAortic atherosclerosis was induced by a combination of endothelial injury and 10 weeks' atherogenic diet. Animals were then randomized to receive the foregoing diet without or with 1% (wt/wt) probucol for 16 weeks. At the end of week 26, in vivo intravascular ultrasound, pathological, immunohistochemical and gene expression studies were performed.

RESULTSProbucol significantly decreased vessel cross-sectional area, plaque area and plaque burden without effect on lumen area. More negative remodeling and less positive remodeling occurred in the abdominal aortas of probucol group than the control group (56% vs. 21%, 18% vs. 54%, respectively, both P < 0.01). In addition, the probucol group showed a smaller mean remodeling index relative to the control group (0.93 ± 0.13 vs. 1.05 ± 0.16, P < 0.01). Furthermore, probucol treatment decreased macrophage infiltration, inhibited apoptosis of cells within plaques, and reduced the production of matrix metalloproteinases-2, -9, cathepsin K and cathepsin S (all P < 0.01).

CONCLUSIONSThese findings suggest that probucol may attenuate the enlargement of atherosclerotic vessel walls and be associated with a negative remodeling pattern without affecting the lumen size. This effect may involve inhibition of extracellular matrix degradation and prevention of apoptosis in atherosclerotic plaques.

Animals ; Anticholesteremic Agents ; pharmacology ; Aorta ; pathology ; Apoptosis ; drug effects ; Atherosclerosis ; drug therapy ; metabolism ; pathology ; Lipids ; blood ; Macrophages ; drug effects ; physiology ; Male ; Matrix Metalloproteinase 2 ; metabolism ; Matrix Metalloproteinase 9 ; metabolism ; Oxidative Stress ; Probucol ; pharmacology ; Rabbits ; Ultrasonography, Interventional ; methods

OBJECTIVETo use the data of occupational epidemiology to estimate the benchmark dose (BMD) of renal dysfunction induced by lead.

METHODSBlood lead was considered as an exposure biomarker, while urinary total protein (TP), urinary beta(2)-microglobulin (beta(2)-MG) and urinary N-Acetyl-beta-D-glucosaminidase (NAG) were considered as effect biomarkers reflecting the damage of renal function. The dichotomized (binary) data was used as effect endpoints. The BMD and BMD lower limit (BMDL) of blood lead were estimated at the 10% benchmark response using BMDS version 1.3.1.

RESULTSThere was an increased prevalence of hyper-TP-uria, hyper-beta(2)-MG-uria and hyper-NAG-uria with an increasing blood lead concentration. There was obviously dose-response relationship between blood lead and TP, beta(2)-MG and NAG, respectively. The BMD and BMDL of blood lead affecting renal function were estimated to be 323.6 - 754.3 microg/L and 274.2 - 541.5 microg/L. The BMDL of blood lead was ranged from low to high as NAG, TP and beta(2)-MG. The urinary NAG activity might be served as a sensitive biomarker in detecting early renal dysfunction.

CONCLUSIONIt should be feasible to use the BMD approach to set up the reference dose (RfD) and reference concentration (RfC). BMD approach might provide a new and better way for setting up the RfD/RfC.

Acetylglucosaminidase ; urine ; China ; epidemiology ; Clinical Chemistry Tests ; methods ; standards ; Humans ; Lead ; blood ; Lead Poisoning ; blood ; epidemiology ; urine ; Occupational Exposure ; analysis ; Prevalence ; Proteinuria ; urine ; beta 2-Microglobulin ; urine

Objective To investigate the effects of uncoupling protein 2 (UCP2) on the myocardial cells of mice with type 2 diabetes mellitus combined with hyperuricemia (HUA), and clarify the mechanism thereof. Methods The mouse cardiac myocytes (MCM) cultured with 25mmol/L high glucose (HG) medium were divided into two groups: HG plus 300μmol/L sodium palmitate for 18 hours as high glucose and high fat (HG+HF) group, and HG+HF plus 1500μmol/L uric acid (UA) for 18 hours as HG+HF+HUA group. Then the myocardial cells in HG+HF+HUA group, by use or not use UCP2 inhibitor genipin, were further divided into two groups: vehicle group and genipin group. In order to verify the mechanism of UCP2 in myocardial cells injury caused by high glucose, high lipid and high uric acid, the myocardial cells were divided again into genipin group and genipin+N-acetylcysteine (NAC) group. Accordingly, the apoptosis of myocardial cells were measured by flow cytometry at specific time, the mRNA and protein expressions of UCP2 were determined by q-PCR and Western blotting, and the levels of reactive oxygen species (ROS) were detected by DHE staining and ELISA. Results The apoptosis rate of myocardial cells increased obviously, and the expression levels of UCP2 decreased and of ROS elevated significantly in HG+HF+HUA group than in HG+HF group (P<0.05). As the expression levels of UCP2 decreased by genipin intervention, the apoptosis rate of myocardial cells and ROS level in HG+HF+HUA group increased more obviously (P<0.05). In contrast, such an effect was reversed by the application of antioxidants NAC (P<0.05). Conclusion UCP2 can inhibit oxidative stress and alleviate the apoptosis of myocardial cells induced by high glucose, high fat and high uric acid.

As one of the most serious types of psoriasis, pathogenesis of erythrodermic psoriasis (EP) is unclear so far. In this study, we aimed to detect the levels of Th1/Th2 cytokine-associated transcription factors and T-lymphocyte clone in peripheral blood mononuclear cells (PBMCs) derived from EP patients, and gene expression level of T-bet/GATA-3 in skin lesion. The potential role of Th1/Th2 reaction pattern played in the pathogenesis of EP was also discussed. Serum levels of IFN-γ, IL-2, IL-4 and IL-10 were quantified by ELISA among 16 EP patients, 20 psoriasis vulgaris (PV) patients and 15 healthy controls. The expression levels of T-bet/GATA-3 in the skin lesion and PBMCs were examined by real-time qPCR. The ratio of Th1/Th2 was measured by flow cytometry. The levels of IFN-γ, IL-2, IL-4 and IL-10 were higher in EP patients than in the healthy controls. The levels of IL-4 and IL-10 were 69.44±11.45 and 12.62±4.57 pg/mL, respectively, in EP patients, significantly higher than those in PV patients and healthy controls (P<0.05). Flow cytometry revealed the levels of both Th1 and Th2 in PBMCs from EP patients were higher than those in healthy controls, and the Th1/Th2 ratio was dramatically lower than in PV patients (P<0.01). The ratios of IFN-γ/IL-4 and T-bet/GATA-3 in EP patients were both less than 1.0, suggesting a reversal when compared with the other two groups. Our study indicated that the EP patients exerted a Th1/Th2 bidirectional response pattern, and the balance of Th cell subsets inclines to Th2, which might be one of the important mechanisms of EP pathogenesis.

Objective:To observe the effects of acupuncture on the characteristics of neuro-electrophysiological activity in hippocampal CA1 and CA3 areas of rats with post-traumatic stress disorder (PTSD).Methods:Fifty Sprague-Dawley (SD) rats were randomly divided into a blank group,a model group,a grasping group,a Western medicine group and an acupuncture group,with 10 rats in each group.Except for the blank group,rats in the other 4 groups all received the combined stress modeling method.Rats in the Western medicine group were intragastrically administrated with paroxetine hydrochloride,those in the acupuncture group received acupuncture intervention,those in the grasping group received grasping fixation,and those in the model group and the blank group did not receive any interventions.After 14 d of intervention,the interspike interval (ISI) and power spectral densities (PSD) were analyzed and mapped by in vivo multiple channels to record the neuron clusters discharge in the hippocampal CA1 and CA3 areas.Results:Compared with the blank group,ISI was prolonged in the CA1 and CA3 areas of the model group and the grasping group,and the concentrated PSD distribution area moved down (P＜0.05 or P＜0.01).Compared with the grasping group,the ISI of the CA1 and CA3 areas in the Western medicine group and the acupuncture group was shortened,and the concentrated PSD distribution area moved up (P＜0.05 or P＜0.01).The ISI and PSD distributions in the CA1 and CA3 areas of the acupuncture group were not statistically different from those in the Western medicine group (both P＞0.05).Conclusion:Both acupuncture and paroxetine hydrochloride can significantly regulate the neuro-electrophysiology activity of hippocampal CA1 and CA3 areas in PTSD rats,which may be one of the mechanisms of acupuncture intervention to promote PTSD recovery.

A quick HPLC-ESI-MS/MS method was established for simultaneous determination of three chemical compositions in Usnea, including usnic acid, diffractaic acid, and ramalic acid. The separation was performed on a chromatographic column of Agilent ZORBAX SB-C, (4.6 mm x 250 mm, 5 µm), and the mobile phase was methanol (0.05% formic acid)-0.05% formic acid solution (4 mmol ammonium acetate), with an isocratic elution at a flow rate of 0.8 ml · min⁻¹. Multiple reaction monitoring scanning mode (MRM) was performed combined with the ion switching technology in positive and negative ion switching mode to apply for the quantitative determination. The calibration curves for the above three compounds were linear in corresponding injection amount. Their average recoveries were 95.0%-105.1%, with RSDs of 1.1%-5.2%. The method was simple, rapid, accurate with high repeatability, which could provide a reference for overcalling evaluation the quality of Usnea.
Chromatography, High Pressure Liquid ; methods ; Hydroxybenzoates ; analysis ; Spectrometry, Mass, Electrospray Ionization ; methods ; Tandem Mass Spectrometry ; methods ; Usnea ; chemistry

OBJECTIVEDisturbances of the synthesis and breakdown of the extracellular matrix of arterial walls have emerged as key features of the atherosclerotic process. We observed the changes of circulating procollagen marker for type III collagen turnover rate, the N-terminal propeptide P III NP and vascular resistance in hypertensive patients treated with various antihypertensive regimens.

METHODA total of 130 light to moderate hypertensive patients were randomly assigned to receive enalapril (group B, n = 43), enalapril + spirolactone (20 mg/d, group A, n = 44) and anti-hypertensive drugs not directly affecting RAAS (calcium antagonist, beta-blocker, group C, n = 43) for 1 year. Target blood pressure is < 130/80 mm Hg.

RESULTSTarget blood pressure was reached in all treated patients and was similar among various groups. Under the same blood pressure controlling precondition, serum P III NP were similar at baseline among various groups and remained unchanged in group B [(3.4 +/- 0.3) microg/L vs. (3.7 +/- 0.3) microg/L, P > 0.05] and significantly decreased in group A [(2.3 +/- 0.2) microg/L vs. (3.8 +/- 0.2) microg/L, P < 0.05] while significantly increased in group C [(3.9 +/- 2.0) microg/L vs. (3.2 +/- 1.5) microg/L, P < 0.05]. Vascular resistance was similar among groups before therapy and all significantly decreased after 1 year antihypertensive therapy and the decrease was more significant in group A [(1064.3 +/- 158.6) dyn.s(-1).cm(-5)] than that in group B [(1200.8 +/- 298.7) dyn.s(-1).cm(-5)] and group C [(1205.1 +/- 206.4) dyn.s(-1).cm(-5)].

CONCLUSIONSpironolactone in conjunction with enalapril is a more favorable antihypertensive regimen in decreasing P III NP and improving vascular resistance than enalapril alone or antihypertensive drug regimens not directly affecting RAAS.

Adult ; Aged ; Angiotensin-Converting Enzyme Inhibitors ; therapeutic use ; Antihypertensive Agents ; therapeutic use ; Biomarkers ; Enalapril ; therapeutic use ; Humans ; Hypertension ; drug therapy ; metabolism ; physiopathology ; Middle Aged ; Procollagen ; blood ; Spironolactone ; therapeutic use ; Vascular Resistance

Relationship between ATP changes of rabbit blood and postmortem interval (PMI) was studied. Twenty-four healthy rabbits were sacrificed and randomly divided into 3 groups with 8 rabbits of each group. The bodies of three groups were placed in calorstat at temperature of 15°C, 25°C and 35°C, respectively. The blood from the right ventricle was sampled through indwelling needle each 4 h until 72 h after death. ATP levels in the blood samples were measured by using ATP fluorescence rapid detection technique at different PMIs. Blood ATP levels slightly increased in the early stage after death and then constantly declined at all temperatures (15°C, 25°C, and 35°C). Cubic polynomial regression equations with log[ATP] as dependent variable (y) and PMI as independent variable (x) at different temperatures and the optimal time period were established as followed: Under 15°C and during 16-64 h after death, y=-3.027×10(-5)x(3)+0.003x(2)-0.096x-10.625 (R a (2)=0.992, P<0.001); under 25°C and during 8-56 h after death, y=-2.921×10(-5)x(3)+0.002x(2)-0.059x-11.186 (R a (2)=0.989, P<0.001); under 35dgC and during 4-36 h after death, y=-9.769×10(-5)x(3)+ 0.005x(2)-0.117x-11.166 (R a (2)=0.991, P<0.001). The changes in ATP levels in blood collected from right ventricle of rabbit cadavers showed relatively stable and regular degradation within 72 h after death at different temperatures.
Adenosine Triphosphate ; blood ; Animals ; Autopsy ; methods ; Body Temperature ; physiology ; Female ; Forensic Pathology ; methods ; Male ; Postmortem Changes ; Rabbits ; Reproducibility of Results ; Sensitivity and Specificity

Alterations of the autophagy-lysosomal pathway (ALP) and autophagy have been involved in lung ischemia-reperfusion (I/R) injury. However, dynamic imaging of ALP function under lung I/R injury particularly is not fully understood. Here we depicted the live-cell fluorescence imaging of autophagosome to monitor ALP activation and autophagy function. The pAsRed2-N1-LC3 vectors were transfected into CRL-2192 NR8383 (an alveolar macrophage cell line) and CCL149 (an alveolar epithelial cell line) successfully. 0-h, 2-h, 4-h, and 6-h hypoxia/0-h, 2-h, 4-h, and 6-h reoxygenation were then induced with an ALP inhibitor (3-MA) or activator (rapamycin) in the culture of transfected cells separately. ALP activation was conformed by up-regulating AMPK and beclin1 expression. Apoptosis was not obvious in 2-h hypoxia/2-h reoxygenation. pAsRed2-N1-LC3 CCL149 and pAsRed2-N1-LC3 NR8383 cells revealed gradually enhanced AsRed2 from 2-h to 6-h hypoxia/reoxygenation. AsRed2 varied sensitively to 3-MA and rapamycin interventions during 2-h hypoxia/reoxygenation. Our data provides a simple method of autophagosome imaging to monitor ALP activation and autophagy function in lung I/R injury.
Animals ; Autophagy ; Base Sequence ; DNA Primers ; Hypoxia ; physiopathology ; In Vitro Techniques ; Lung ; physiopathology ; Lysosomes ; physiology ; Oxygen Inhalation Therapy ; Rats ; Real-Time Polymerase Chain Reaction