Objective:To explore the safety and efficacy of ultrasound-assisted facial filler injection, based on the anatomy of facial vessels to prevent intravascular embolization.Methods:From Jan. 2019 to Sep. 2020, 142 patients were treated with facial soft-tissue filler injection (mean age, 39.7 years; 131 female and 11 male). According to the patients＇ own will, autologous fat or hyaluronic acid was applied respectively. When injecting, the assistant could press over the periorbital artery to temporarily occlude the artery, confirmed with Doppler ultrasound, thus reduced the risk of intravascular embolization, and carefully injected with minimal pressure and tiny amount.Results:A total of 142 patients were enrolled in the study, and 54 patients were treated with autologous fat grafting, while 88 patients were injected with hyaluronic acid. The injection sites included forehead, temple, glabella, nasal root, tear trough, nasolabial fold, cheek, chin, and lips. Facial rejuvenation improvement was satisfied with a smooth contour and proper augmentation. No vascular embolization occurred. 9 patients received a second or third round fat grafting to achieve better outcome. Follow-up duration ranged from 1 month to 6 months.Conclusions:With ultrasound assistant, digital pressure over the orbital artery could temporarily occlude the artery and may reduce the risk of intravascular embolization. The simple technique may add a significant benefit with no additional cost or risk to the patients.

To develop a new recombinant hepatitis E vaccine, we used Hansenula polymorpha expression system to express recombinant hepatitis E virus-like particles (HEV VLPs), to construct a recombinant engineered strain HP/HEV2.3. The fermentation conditions and purification process were studied next. The first working seed lots were fermented in liquid culture, and the fermentation products were collected, then crushed, clarified, purified by ultrafiltration, silica gel adsorbed and desorbed, concentrated by ultrafiltration, purified by liquid chromatography and sterilized by filtration. The purity reached 99% with a yield of 33%. Electron microscopy analysis revealed that both the purified recombinant HEV VLPs from HP/HEV2.3 and natural hepatitis E virus particles appear identical of being 32 nm. The resulting DNA sequence obtained from VLPs is identical to the published HEV sequence. The SDS-PAGE analysis has revealed that the protein molecular weight of the HEV VLPs is 56 kDa, and the expression product HEV VLPs were accumulated up to 26% of total cellular protein. The expression level is 1.0 g/L. Western blotting, enzyme-linked immunosorbent assay (ELISA) results of the protein and ED₅₀ of the vaccine showed that the HEV VLPs have good antigenicity and immunogenicity. In summary, the recombinant HEV VLPs from Hansenula polymorpha can be used in the manufacture of a new genetically engineered vaccine against hepatitis E.