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Objective To optimize the preparation of high-efficiency galactocylated poly-L-lysine (Gal-PLL) ligand of the asialoglycoprotein receptor in liver, providing premise and foundation for upper preparation of ultrasound contrast agent of liver targeted nanoscale perfluorocarbon microballoon and the liver targeted molecular imaging. Methods Chemical reactions of reductive amination were carried out on group A and group B according to different proportions of reaction component. Each group was subdivided into three subgroups. In group A, three different molar ratios of D-galactose and poly-L-lysine (PLL) were compounded respectively with equivalent and sufficient reductant borohydride. In group B, identical molar ratios of D-galactose and PLL were compounded respectively with three unequal reductants borohydride. Products of each group were separated and purified by sephadex column to acquire different molecular weight distributions and the results were analyzed. Results In the condition of identical reductant, the peak curve of compound's molecular weight appeared earlier when D-galactose decreased properly. In the condition of identical molar ratio of D-galactose and PLL,the peak curve of compound's molecular weight appeared also earlier when reductant decreased properly. When the molar ratio of D-galactose and reductant was 1∶1, the peak curve of compound Gal-PLL and free components was more obvious, and the quantity of compound Gal-PLL increased to maximum. Conclusions In the condition of identical reductant, coupling effect of D-galactose and PLL increased when D-galactose decreased properly. In the condition of identical molar ratio of D-galactose and PLL, coupling effect was better when reductant decreased properly. When the molar ratio of D-galactose and reductant was 1∶1, coupling effect of them was the best. The coupling of D-galactose and PLL was related to not only the proportion of D-galactose and PLL, but also the proportion of D-galactose and reductant.
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Objective To prepare the liver targeting nano-liquid perfluorocarbon ultrasound contrast agent and observe its general characteristics;to observe the targeting combined effects of the human liver cells L02 and the targeted ultrasound contrast agent ;to evaluate the gathering imaging effects of the targeted ultrasound contrast agent. Methods Amine method was used to prepared asialoglycoprotein Gal specific ligand polylysine (Gal-PLL), rotary evaporator and sonicated liquid fluorocarbon were used to prepare nano lipid ultrasound contrast agent. Human liver cell L02 were cultured, the combined effects were observed according to the reacting time of the cells and the targeted nano-lipid ultrasound contrast agent. The nanolipid ultrasound contrast agent and the degassed_ water_ were loaded into cysts and their ultrasound imaging effects were observed by ultrasound diagnostic apparatus Philips iU22. Results The particle size of the liquid fluorocarbon nano-targeted lipid ultrasound contrast agent was extremely small, uniform, cylindrical and spherical. The cysts in vitro showed that the side of the targeted liquid perfluorocarbon nano-lipid ultrasound contrast agent showed high echo. Conclusions The targeted liquid perfluorocarbon nano-lipid ultrasound contrast agent can be effectively targeted to the human liver cells L02 due to carrying home-made Gal-PLL. The targeted ultrasound contrast agents can be imaging by ultrasound and be confirmed in vitro.The size of the contrast agent was small, therefore, it can be considered an ideal ultrasonic molecular probe and achieve the ultrasound molecular imaging in cell level.
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Objective To explore the impact of nucleos(t)ide analogue antiviral therapy on anxiety and depression of patients with chronic hepatitis B (CHB),and analyze the related factors.Methods Before nucleos(t)ide analogue antiviral therapy,1 year and 2 years after antiviral therapy,120 CHB patients were investigated with self-rating anxiety scale (SAS) and self-rating depression scale (SDS).The demography data of patients were collected.Serum levels of alanine transaminase (ALT) and hepatitis B virus (HBV) DNA and other biochemical indicators were measured regularly.Results Before nucleos(t)ide analogue antiviral therapy,1 year and 2 years after antiviral therapy,both the mean scores of SAS and SDS became lower gradually (F=12.661 and 22.395,respectively;both P＜0.01).The percentage of patients with SAS and SDS scores more than 50 were 5.8％,4.2％,1.7％ and 13.3％,7.5％,5.0％,respectively.After 2 years of therapy,the anxiety improvement rate of the patients obtained HBV DNA＜1000 copy/mL was 69.0％,while those with HBV DNA≥1000 copy/mL was 22.2％ (x2 =22.325,P＜0.01).Meanwhile,after 2 years of therapy,the depression improvement rate of the patients obtained HBV DNA＜1000 copy/mL was 77.4％,while those with HBV DNA≥1000 copy/mL was 22.2％ (x2 =32.179,P＜0.01).Multiple factors Logistic regression analysis indicated that the odds ratios (OR) of improvement of anxiety and depression in patients with HBV DNA＜1000 copy/mL were 7.751 (95％ CI:3.026-19.853) and 15.069(95％ CI:5.309-42.770),respectively,compared with those with HBV DNA≥1000 copy/mL; and OR of improvement of depression in patients with ALT≤40 U/L waa 4.103 (95％ CI: 1.376 - 12.238).Conclusions Nucleos(t) ide analogue antiviral therapy could improve the anxiety and depression of CHB patients.The HBV DNA negativity is the independent impact factor of improvement of anxiety and depression in CHB the patients.
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Objective To study the feasibility of the Fe3O4-loaded lipid perfluorooctylbromide nanoparticles (Fe3O4-PFOB) for enhanced ultrasound imaging.Methods The Fe3O4-PFOB nanoparticles,incubated with RAW264.7 macrophage cells,were monitored by microscope and ultrasound.Twelve SD rats were randomized into two groups,Fe3O4-PFOB group and PFOB group.Ultrasound imaging of rats' liver was performed before and after intravenous injection of the contrast agents.The liver echogenic intensity was quantified by DFY ultrasound quantified system analysis.Results Incubation of the Fe3O4-PFOB nanoparticles with macrophages resulted in the uptake of Fe3O4-PFOB by macrophages.Macrophages loaded with Fe3O4-PFOB exhibited enhanced echogenicity in vitro.In in vivo imaging,Fe3O4-PFOB produced better and prolonged ultrasound enhancement of rats' liver compared to PFOB nanoparticles.Conclusions Fe3O4-PFOB nanoparticles could enhance ultrasound imaging and may potentially serve as a multimodal probe for ultrasound,CT and MR imaging.
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Objective To investigate the targeted capacity of ultrasound contrast agent with a galactosepoly-L-lysine ligand,and lay the foundation for targeted imaging in vivo and the treatment of liver cancer.Methods The small molecular targeted ligand was synthetized by a method of reductive amination,the targeted lipid microbubbles were performed by combining lipid microbubbles with the targeted ligand,the targeted effectiveness was observed by fluorescence microscopy.Results Galactose and poly-L-lysine were combined effectively after 24 h with the molar ratio 1:1 00,the average particle size of the targeted lipid microbubbles was 2 micron,the targeted combine of the HepG2 liver cancer cells with the targeted ultrasound contrast agent was observed by laser scanning confoeal microscope.Conclusions The targeted lipid miembubbles can target effectively to the HepG2 cells.