In recent years, Hepatitis B virus (HBV) persistent infection mouse model with recombinant adeno-associated virus 8 carrying 1.3 copies of HBV genome (rAAV8-1.3HBV) is concerned. We studied and compared the efficacy among HBV persistent infection mice models by other serotypes except AAV8. First, we prepared and purified five viruses: rAAV1-1.3HBV, rAAV2-1.3HBV, rAAV5-1.3HBV, rAAV8-1.3HBV and rAAV9-1.3HBV. Then we injected each virus into 3 C57BL/6J mice with the dose of lx 1011 vg (Viral genome, vg) per mouse. We detected HBsAg and HBeAg in sera by enzyme-linked immunosorbent assay (ELISA) at different time points post injection. We killed mice 8 weeks post injection and took blood and livers for assay. We detected copies of HBV DNA by real-time quantitative PCR in sera and livers. Meantime, we detected HBcAg in the livers of mice by immunohistochemistry and further performed pathology analysis of these livers. The five groups of mice, HBeAg and HBsAg expression sustained 8 weeks in serological detection and HBV DNA was both detected in sera and livers at the time of 8 weeks post injection. HBeAg, HBsAg, HBV DNA copies expression levels in descending order were AAV8>AAV9>AAV1>AAV5>AAV2. HBcAg expression was detected in livers as well. Varied degrees of liver damage were shown in five groups of mice. This study provides more alternative AAV vector species to establish a persistent infection with hepatitis B model.
Animals ; Dependovirus ; classification ; Disease Models, Animal ; Enzyme-Linked Immunosorbent Assay ; Genetic Vectors ; Genome, Viral ; Hepatitis B ; virology ; Hepatitis B Core Antigens ; metabolism ; Hepatitis B Surface Antigens ; blood ; Hepatitis B e Antigens ; blood ; Hepatitis B virus ; genetics ; Mice ; Mice, Inbred C57BL ; Serogroup ; Virus Replication

Objective:To investigate the clinical effect of autologous nano-fat injection to correct upper eyelid depression.Methods:From March 2016 to March 2020, a total of 191 eyes were treated in 110 patients with upper eyelid depression, including 81 cases on both sides and 29 cases on one side. The fat was obtained by negative pressure liposuction from the thigh. After the nano treatment, the obtained nano fat was injected into the subcutaneous layer, ROOF layer and orbital septum evenly with multi-tunnel without tension.Results:Among 110 patients, 1-2 ml of nano-fat was filled to the upper eyelid unilaterally, with an average of 1.5 ml. 95 patients were followed up for 2-24 months, with average 6 months. After one injection, the depression of the upper eyelid was significantly improved in all patients, and the correction of the depression of the upper eyelid was insufficient in 28 patients. After 3 months, autologous nano fat filling was performed again, and the results were satisfactory. Temporary ptosis occurred in 5 patients and recovered naturally after 3-6 months. Local ecchymosis and swelling were observed in 12 patients, and resolved spontaneously within 7-14 days. No infection, nodules or other complications occurred in all the patients.Conclusions:Nano-fat is a safe and effective method, which can effectively fill the orbital depression area and correct the depression of upper eyelid, with less adverse reactions.