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Objective@#To investigate the feasibility of echocardiography-guided closed-chest repeated intraventricular blood sampling in mice, and to clarify the maximum blood volume that can be collected by this method, and whether the method can be used for long-term repeated blood collection in mice.@*Methods@#Twenty-four male C57BL/6J mice (10-14 weeks old) were divided into the terminal experiment group (n=4, for investigating the maximum blood amount that could be sampled at one time), the repeated 0.5 ml blood collection group (n=10, sampling 0.5 ml whole blood each time, once every two days for consecutive 4 weeks), and the repeated 0.75 ml blood collection group (n=10, sampling 0.75 ml whole blood each time, once every two days for consecutive 4 weeks). High-frequency echocardiography was used to display the largest section of the left ventricle, guiding the insulin syringe needle through the thorax into the left ventricle for blood collection. In the repeated 0.5 ml blood collection group, echocardiography was used to detect the cardiac structure and function before blood collection, three minutes after blood collection, and one week after the last (the 14th) blood collection.@*Results@#We successfully performed echocardiography-guided closed-chest intraventricular blood sampling, with an average operating time (88±19)s per mouse, and a maximum blood volume (1.43±0.11)ml per mouse. In the repeated 0.5 ml blood collection group, heart rate, left ventricular ejection fraction, left ventricular fractional shortening, left ventricular end-diastolic dimension and left ventricular posterior wall end-diastolic thickness remained uncganged before the first blood collection and after 4 weeks of repeated blood collection (all P>0.05). No death in the repeated 0.5 ml blood collection group. However, in the 0.75 ml blood collection group, two mice died before the end point.@*Conclusions@#The echocardiography-guided closed-chest intraventricular blood sampling is a safe, minimally invasive, convenient and efficient method, and can be used repeatedly for long-term blood collection in mice.
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This study investigated the effects of acidity and metal ion on the antibacterial activity of α- and ß-chitosan at different molecular weights (Mw, 22-360 kDa) against Escherichia coli and Listeria innocua through agar well diffusion assay. Spectrophotometric, electrophoretic, and confocal fluorescence microscopy analysis were further employed to evaluate the antibacterial mechanisms probably involved. Increasing pH from 4.0 to 5.0 weakened the antibacterial ability of chitosan as shown by the decreased bacteria growth inhibition zone (BGIZ) from 0.63 to 0.57 cm for ß-chitosan (61 kDa) and from 0.62 to 0.57 cm for α-chitosan (30 kDa) against E. coli. All ß- and α-chitosan samples showed antibacterial activity against L. innocua, in which 22 kDa ß-chitosan and 30 kDa α-chitosan at pH 4.0 had the highest antibacterial activity with BGIZ of 1.22 and 0.98 cm, respectively. Interactive effect between pH and Mw on the antibacterial activity of ß-chitosan was observed, but not of α-chitosan. Adding Co(2+) and Ni(2+) significantly improved the antibacterial activity of chitosan, while adding K(+), Na(+), and Li(+) significantly weakened the antibacterial activity of some ß- and α-chitosan samples (P < 0.05), and different Mw and forms of chitosan showed different metal ion absorption capacities. Results indicate that chitosan might insert into the groove of bacterial DNA double helix structure to induce DNA degradation and permeate through bacteria cell membranes and combine with genomic DNA to induce its dysfunction, providing evidences for the antibacterial mechanisms of chitosan.
