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H9N2 and H3N2 are the two most important subtypes of low pathogenic avian influenza viruses (LPAIV) because of their ongoing threat to the global poultry industry and public health. Although commercially available inactivated H9N2 vaccines are widely used in the affected countries, endemic H9N2 avian influenza remains uncontrolled. In addition, there is no available avian H3N2 vaccine. Influenza virus-like particles (VLPs) are one of the most promising vaccine alternatives to traditional egg-based vaccines. In this study, to increase the immunogenic content of VLPs to reduce production costs, we developed chimeric bivalent VLPs (cbVLPs) co-displaying hemagglutinin (HA) and neuraminidase (NA) of H9N2 and H3N2 viruses with the Gag protein of bovine immunodeficiency virus (BIV) as the inner core using the Bac-to-Bac baculovirus expression system. The results showed that a single immunization of chickens with 40μg/0.3mL cbVLPs elicited an effective immune response and provided complete protection against H9N2 and H3N2 viruses. More importantly, cbVLPs with accompanying serological assays can successfully accomplish the strategy of differentiating infected animals from vaccinated animals (DIVA), making virus surveillance easier. Therefore, this cbVLP vaccine candidate would be a promising alternative to conventional vaccines, showing great potential for commercial development.

Aim: To test the ability of carotid stiffness evaluated by using ultrafast ultrasound imaging to indicate coronary atherosclerosis and its association with the severity of coronary artery disease (CAD).Material and Methods: This cross-sectional study included 131 patients with CAD and 60 normal controls. Carotid intima-media thickness (cIMT) was measured by two-dimensional ultrasound. Carotid stiffness was determined by ultrafast ultrasound imaging, with which the carotid pulse wave velocity at the beginning (PWVBS) and end (PWVES) of systole were calculated. Gensini scores based on coronary angiography were used to estimate the severity of CAD.Results: Compared with normal controls, the CAD patients had higher carotid diameters, cIMT, PWVBS and PWVES (p < 0.05). In the CAD group, Gensini scores correlated significantly with cIMT, PWVBS and PWVES (r = 0.279, p = 0.001; r = 0.661, p < 0.001; r = 0.620, p < 0.001; respectively). The multivariate analysis further indicated that PWVBS, PWVES and body mass index were independently associated with the Gensini score (β = 0.466, p < 0.001; β = 0.308, p < 0.001; and β = 0.209, p = 0.001; respectively). In addition, the sensitivity and specificity were 54% and 83%, respectively, for PWVBS (cutoff value, 6.9 m/s; area under the receiver operating characteristic curve, 0.70) and 64% and 83%, respectively, for PWVES (cutoff value, 8.0 m/s; area under the curve, 0.73).Conclusions: Increased carotid PWVBS and PWVES detected by ultrafast ultrasound imaging as indices of carotid stiffness might serve as promising indicators for CAD and its severity. [ABSTRACT FROM AUTHOR]