[Preparation of an anti-cotinine monoclonal antibody and its application in immunological detection].

OBJECTIVE: To prepare monoclonal antibody against cotinine (COT) and to establish immunoassay for detecting COT in human urinary samples. METHODS: BALB/c mice were immunized with synthesized cotinine-bovine serum albumin (COT-BSA) to screen monoclonal antibody with technique of cell fusion. The monoclonal antibody was used for the indirect competitive enzyme-linked immunosorbent assay (ic-ELISA) and colloidal gold immunochromatographic strip assay for the detection of COT in human urine. RESULTS: The monoclonal antibody against COT was identified by ic-ELISA with a 50%inhibitive concentration (IC50) value of 21 ng/mL; and it was also identified by colloidal gold immunochromatographic strip assay with a cut-off value of 100 ng/mL. For ic-ELISA, the range of detection was 0-100 ng/mL with a minimal limit of 0.1 ng/mL; the recovery of assay was 99.41%-117.98%, and the intra-assay and inter-assay coefficient variations were not higher than 15.31%and 15.07%, respectively. For colloidal gold immunochromatographic strip assay, the accuracy of stability and repeatability both were 100%. CONCLUSIONS: The ic-ELISA and colloidal gold immunochromatographic strip assay using the prepared monoclonal antibody against COT have been proved to be reliable for the rapid detection of COT in human urines, which may be used for monitoring of environmental tobacco smoke.

U0126 attenuates ischemia/reperfusion-induced apoptosis and autophagy in myocardium through MEK/ERK/EGR-1 pathway.

Myocardial ischemia is one of the main causes of sudden cardiac death worldwide. Depending on the cell type and stimulus, ERK activity mediates different anti-proliferative events, such as apoptosis, autophagy, and senescence. The aim of this study was to determine the protective effect of 1,4-diamino-2,3-dicyano-1,4-bis[2-aminophenylthio] butadiene (U0126), an ERK kinase inhibitor, on myocardial ischemia/reperfusion (I/R) injury and the mechanisms involved. An I/R model was established in vivo in C57BL/6 mice and in vitro using mouse cardiomyocytes, respectively. To evaluate the protective effects of U0126 on I/R injury, we measured the myocardial infarct area, apoptosis, and autophagy. Our data indicated that pretreatment with U0126 significantly reduced the infarct area caused by I/R. Moreover, U0126 reduced the caspase-3 activity and the number of TUNEL-positive cardiomyocytes, which together indicate decreased apoptosis. Additionally, U0126 remarkable reduced the level of Beclin-1 and LC3 and increased p62 expression, which indicates that U0126 suppressed H/R-induced autophagy. Furthermore, the relationship between U0126 and MEK/ERK pathway activation in H/R-induced cardiomyocytes was also investigated. U0126 ameliorated H/R injury through inhibition of the MEK/ERK pathway and by suppressing in the downstream EGR-1 expression. Together, our research suggests that U0126 may protect against H/R injury by preventing H/R-induced myocardium apoptosis and autophagy via the MEK/ERK/EGR-1 pathway, and may be a potential therapeutic approach for attenuating myocardial I/R injury.

Asymptomatic polyvascular abnormalities in community (APAC) study in China: objectives, design and baseline characteristics.

OBJECTIVE: The population-based "Asymptomatic Polyvascular Abnormalities in Community (APAC) Study was designed to examine prevalence and associations of asymptomatic polyvascular abnormalities (APA) in a general population. In this report, the objectives, design and baseline data of the APAC study are described. METHODS: The study included 5,440 participants (40.1% women) with an age of 40+ years who were randomly selected from the population of the Kailuan Study which included 101,510 employees and retirees of the Kailuan Co. Ltd, a large coal mine industry located in Tangshan, Hebei, China. Exclusion criteria were previous cerebral stroke, transient ischemic attacks and coronary heart disease. In 2010 and 2011, information on potential cardiovascular risk factors was collected and all participants underwent transcranial Doppler sonography, measurement of the ankle brachial index, and bilateral carotid duplex sonography. In a first follow-up examination in 2012/2013, retinal photography and spectral-domain optical coherence tomography were additionally performed. In a planned long-term follow-up, data from clinical examinations and laboratory tests and the occurrence of cardiovascular or cerebrovascular events will be collected to build up a predicting model for the risk of ischemic events. RESULTS: At baseline, mean age of the participants was 55.2 ± 11.8 years, and men showed a significantly (P<0.001) higher prevalence of arterial hypertension (55.5% vs. 36.5%) and hyperlipidemia (50.7% vs. 46.0%) and a higher blood homocysteine concentration (18.68 ± 10.28 µmol/L versus 11.69 ± 6.40 µmol/L). CONCLUSIONS: The APAC is the first study to prospectively evaluate the relationship between intracranial arterial stenosis, retinal nerve fiber layer changes, retinal microvascular signs, and the eventual development of cerebrovascular or cardiovascular events.

Prevalence of ideal cardiovascular health and its relationship with the 4-year cardiovascular events in a northern Chinese industrial city.

BACKGROUND: The American Heart Association Committee recently developed definitions of "ideal," "intermediate," and "poor" cardiovascular health based on 7 cardiovascular disease (CVD) risk factors or health behaviors. This study evaluated the prevalence of "ideal" American Heart Association cardiovascular health metrics from June 2006 to October 2007 in the Kailuan cohort (n=101 510; age 18-98 years) in northern China and its relationship with the 4-year CVD incidence. METHODS AND RESULTS: We used Cox proportional hazards regression to calculate hazard ratios and 95% confidence intervals for baseline health behaviors and risk factor categories. The majority of participants (63,676; 69.45%) presented with ≤3 ideal cardiovascular health metrics, whereas 8342 participants (9.1%) had 5 to 7 ideal metrics. Only 93 of 91,698 participants (0.1%) had all 7 metrics in the ideal range. There was a strong relationship between the cumulative incidence of CVD events in the 4-year follow-up and the number of ideal health metrics at baseline; the 1111 participants with 6 and 7 ideal metrics had a significantly lower cumulative incidence of CVD than subjects with no or only 1 ideal health metric (0.8% versus 3.3%). Men had higher rates of CVD events than women (2.46% versus 1.18%). CONCLUSIONS: Few adults had ideal cardiovascular health according to the modified American Heart Association definition. We detected a strong inverse relationship between the cumulative CVD incidence and the number of ideal health metrics at baseline. Population-wide prevention, especially lifestyle improvement, is critical to increase the low-risk prevalence and thereafter decrease CVD events. Clinical Trial Registration- URL: http://www.chictr.org/cn/proj/show.aspx?proj=1441. Unique identifier: ChiCTR-TNC-11001489.