Human alphaherpesvirus 2 (HSV-2) is the main pathogen resulting human genital herpes, which poses a major threat to the socio-economic development, while there is no effective vaccine. In this study, we developed a novel lipopolyplex (LPP)-delivered mRNA vaccine expressing the HSV-2 envelope glycoprotein gD and evaluated its immunogenicity in mice. The mRNA vaccine was prepared from the genetically modified gD mRNA synthesized in vitro combined with the LPP delivery platform and it was named gD-ORI mRNA. The expression of gD antigen in the mRNA vaccine was validated in vitro by Western blotting and indirect immunofluorescence assay, then the immune responses induced by this mRNA vaccine in mice were evaluated. The immunization with gD mRNA alone induced strong humoral and cellular immune responses in mice. Robust and long-lasting gD-specific IgG antibodies were detected in the mouse serum after booster immunization with gD-ORI mRNA. The immunized mice exhibited a Th1/Th2 balanced IgG response and robust neutralizing antibodies against HSV-2, and a clear dose-response relationship was observed. The gD-specific IgG antibodies were maintained in mice for a long time, up to 18 weeks post-booster immunization. At the same time, multifunctional gD-specific CD4⁺ and CD8⁺ T cells in vaccinated mice were detected by intracellular cytokine staining (ICS). This novel gD-expressing mRNA vaccine delivered by LPP induces strong and long-lasting immune responses in mice post booster immunization and has a promising prospect for development and application. This study provides scientific evidence and reference for the development of a new mRNA vaccine for HSV-2.
Animals ; Herpesvirus 2, Human/genetics* ; Viral Envelope Proteins/genetics* ; Mice ; Herpes Genitalis/immunology* ; RNA, Messenger/immunology* ; Female ; Mice, Inbred BALB C ; Antibodies, Viral/blood* ; mRNA Vaccines/immunology* ; Antibodies, Neutralizing/blood* ; Humans

Objective:To construct a novel respiratory syncytial virus (RSV) vaccine based on a recombinant influenza virus vector and evaluate its immune protective effects in mice.Methods:A recombinant H1N1 influenza A virus (IAV) expressing the extracellular domain (Gecto) of RSV A2 G protein was constructed and rescued, named as PR8NAGecto/WSN. After in vitro verification of the Gecto expression and PR8NAGecto/WSN growth kinetics, a single dose of PR8NAGecto/WSN was used to immunize BALB/c mice through intranasal administration to evaluate the efficacy of PR8NAGecto/WSN by assessing humoral (IgG, neutralizing antibody), mucosal (IgA) and cellular immunity (IFN-γ ELISPOT). Four weeks after immunization, the mice were challenged with RSV A2 or RSV B9320 to evaluate the protective effects of PR8NAGecto/WSN by analyzing mouse body weight changes, lung tissue virus titers and pathological changes. Results:A single-dose intranasal immunization with PR8NAGecto/WSN induced robust humoral, mucosal and cellular immunity in mice. Moreover, the mice in the immunized group had lower lung virus loads and mild lung pathological damages following the challenge with RSV A or RSV B subtype as compared with the control group.Conclusions:A single-dose intranasal immunization with PR8NAGecto/WSN induces robust immunity and provide protection against RSV A and B challenges in mice. This study provides new ideas and reference for the development of novel mucosal vaccines against RSV.

OBJECTIVE To evaluate the intervention effect of Caulis sinomenii compatible with prepared Aconiti Lateralis on bone destruction in rheumatoid arthritis （RA）model rats ，and to investigate its mechanism. METHODS Totally 40 SD rats were randomly divided into blank group ，model group ，positive control group （indomethacin 0.013 5 g/kg）and C. sinomenii compatible with prepared Aconiti Lateralis group （C. sinomenii 1.08 g/kg+prepared Aconiti Lateralis 1.35 g/kg）according to body mass ，with 10 rats in each group. Except for the blank group ，all the other groups made RA rat models by injecting type Ⅱ bovine collagen. Rats in each group were given corresponding drugs or distilled water intragastrically. The general information ，body weight ，foot swelling and arthritis index （AI）scores of rats in each group were recorded. After the 30th day of administration ，the changes of ankle bone in rats were detected by small animal CT machine. The levels of inflammatory factors [interleukin- 31（IL-31），IL-25 and IL- 3] and chemokines [receptor activator of nuclear factor κB ligand（RANKL），receptor activator of nuclear factor κB （RANK）and osteoprotegerin （OPG）] in serum were detected by enzyme-linked immunosorbent assay. Pathological indexes of rat ankle joint were observed by HE staining. Immunohistochemical method was used to detect the expression of RANKL ，RANK and OPG in synovial tissue of rat ankle joint. RESULTS Compared with blank group ，the mental state of the model group was weak ， the activity decreased significantly ，the hair lost luster ，and the body weight decreased significantly on the 12th to 30th days （P＜ 0.05 or P＜0.01）；the swelling degree of the foot was significantly increased and the AI score was significantly increased on the 12th to 30th days（P＜0.01）；the ankle joint in model group had rough surface ，obvious tissue damage and serious bone erosion ； serum levels of IL- 31，IL-25，IL-3，RANKL and RANK were increased significantly ，while the level of OPG was decreased significantly （P＜0.01）； the expression of RANKL and RANK in synovium of ankle joint increased significantly ， while the expression of OPG decreased significantly （P＜0.01）. Compared with model group ，the above indexes of administration groups were improved to varying degrees ，and most of the differences were statistically significant （P＜0.05 or P＜0.01）. CONCLUSIONS By inhibiting the RANKL/RANK/OPG signaling pathway ，C. sinomenii compatible with prepared Aconiti Lateralis can inhibit the excessive proliferation of osteoclasts and restore the balance of bone metabolism so as to play a role in protecting bone joints and treating RA.

【Objective】 Compare the early outcome and safety of endoscopy-unilateral laminectomy for bilateral decompression （Endo-ULBD） and posterior lumbar interbody fusion （PLIF） in the treatment of multi-segment lumbar central spinal stenosis. 【Methods】 We retrospectively analyzed 68 patients with multi-segment central lumbar spinal stenosis treated between October 2019 and October 2020 in the Department of Spine Surgery， Affiliated Hospital of Qingdao University. Of them 33 patients were treated with Endo-ULBD and 35 ones were treated with PLIF. We compared the operation time， times of intraoperative fluoroscopy， estimated intraoperative blood loss， incision length， postoperative time to get out of bed， postoperative hospital duration， complications， visual analogue scale （VAS）， Oswestry dysfunction index （ODI） score before and 1 day， 1 month， and 3 months after operation， Japanese Orthopedic Association Assessment Treatment Score （JOA）， and modified MacNab score 3 months after operation between the two groups of patients. 【Results】 Compared with PLIF group， Endo-ULBD group had significantly shorter operation time， smaller incision length， less intraoperative blood loss， shorter postoperative bed time and postoperative hospital stay， and fewer surgical complications （all P<0.05）. There was significantly more intraoperative fluoroscopy in Endo-ULBD group than in PLIF group （P<0.05）. The VAS， ODI and JOA scores of the two groups were significantly improved after treatment （P<0.05）. There was no statistical difference in VAS of leg pain between the two groups after treatment （P>0.05）. However， after treatment Endo-ULBD group outperformed PLIF group in lower back pain VAS， ODI， JOA and the 3-month follow-up excellent and good rates （P<0.05）. 【Conclusion】 For patients with multi-segment central lumbar spinal stenosis， Endo-ULBD treatment can achieve better early clinical outcome than PLIF surgery， with less bleeding， shorter operation time， faster postoperative recovery， and fewer complications.