The aim of this study is to explore the immunogenicity of African swine fever virus p37 recombinant protein in mice.C57BL/6J mice were immunized subcutaneously in the abdomen using p37 recombinant protein as antigen.The second immunization was performed 21 d after the first immunization.Serum-specific antibody levels were detected by ELISA;serum cytokine levels were detected using a multifactor assay technique;mice splenic lymphocytes were isolated 7 d after sec-ondary immunization,and the number of splenic lymphocytes secreting IFN-γ after recombinant protein stimulation was detected by ELISpot;and the ratio of CD4+T cells to CD8+T cells was detected by flow cytometry.The results of indirect ELISA showed that p37 recombinant protein could stimulate mice to produce high levels of specific antibodies;ELISpot showed that p37 recom-binant protein could significantly stimulate splenic lymphocytes to produce IFN-γ(P＜0.001)and activate cellular immune responses;the results of flow cytometry showed that it could signifi-cantly stimulate the differentiation of T-lymphocytes to CD4+T-lymphocytes(P＜0.001).In ad-dition,serum levels of IL-2,IL-4,IFN-γ,and TNF-α immune-related cytokines were significantly higher after the second immunization.Immunization of mice with p37 recombinant protein induced strong humoral and cellular immune responses with good immunogenicity,providing reference for the subsequent epitope identification and functional study of p37 protein and the antigen screening of ASF mRNA vaccine.

To explore the interaction between ASFV capsid protein M1249L and host from the host cellular perspective,M1249L was selected for constructing the bait plasmid(pGBKT7-M1249L)to screen the bone marrow-derived macrophages(BMDMs)cDNA library.After again co-transform and sequence alignment,20 candidate interacting host proteins were screened,such as IL-1β,CTSB and DNAJA3.And then,co-immunoprecipitation assay was performed to verify the interaction be-tween M1249L and host proteins.GO ontology(GO)and KEGG pathway enrichment analyses re-vealed that biological regulation,cellular communication and response to stimulus and others were enriched in biological processes.And these host proteins could share some pathways,including toll-like receptor signaling pathway and Nod-like receptor signaling pathway.Therefore,the results provides the theoretical basis for further research on the mechanism of ASFV M1249L in viral in-fection and immune regulation.

The aim of this study is to explore the immunogenicity of African swine fever virus p37 recombinant protein in mice.C57BL/6J mice were immunized subcutaneously in the abdomen using p37 recombinant protein as antigen.The second immunization was performed 21 d after the first immunization.Serum-specific antibody levels were detected by ELISA;serum cytokine levels were detected using a multifactor assay technique;mice splenic lymphocytes were isolated 7 d after sec-ondary immunization,and the number of splenic lymphocytes secreting IFN-γ after recombinant protein stimulation was detected by ELISpot;and the ratio of CD4+T cells to CD8+T cells was detected by flow cytometry.The results of indirect ELISA showed that p37 recombinant protein could stimulate mice to produce high levels of specific antibodies;ELISpot showed that p37 recom-binant protein could significantly stimulate splenic lymphocytes to produce IFN-γ(P＜0.001)and activate cellular immune responses;the results of flow cytometry showed that it could signifi-cantly stimulate the differentiation of T-lymphocytes to CD4+T-lymphocytes(P＜0.001).In ad-dition,serum levels of IL-2,IL-4,IFN-γ,and TNF-α immune-related cytokines were significantly higher after the second immunization.Immunization of mice with p37 recombinant protein induced strong humoral and cellular immune responses with good immunogenicity,providing reference for the subsequent epitope identification and functional study of p37 protein and the antigen screening of ASF mRNA vaccine.

To explore the interaction between ASFV capsid protein M1249L and host from the host cellular perspective,M1249L was selected for constructing the bait plasmid(pGBKT7-M1249L)to screen the bone marrow-derived macrophages(BMDMs)cDNA library.After again co-transform and sequence alignment,20 candidate interacting host proteins were screened,such as IL-1β,CTSB and DNAJA3.And then,co-immunoprecipitation assay was performed to verify the interaction be-tween M1249L and host proteins.GO ontology(GO)and KEGG pathway enrichment analyses re-vealed that biological regulation,cellular communication and response to stimulus and others were enriched in biological processes.And these host proteins could share some pathways,including toll-like receptor signaling pathway and Nod-like receptor signaling pathway.Therefore,the results provides the theoretical basis for further research on the mechanism of ASFV M1249L in viral in-fection and immune regulation.

African swine fever(ASF)is a highly contagious disease caused by the African swine fe-ver virus(ASFV).To evaluate the immunogenicity of the pp62(CP530R)protein of ASFV,the recombinant CP530R protein was expressed in HEK 293F cells transfected with the plasmid pMAL-Fc-CP530R.Six-week-old female C57BL/6J mice were immunized with 10 μg of purified pp62 protein via subcutaneous injection,followed by a booster immunization with the same dosage at 21 days(enhanced).The humoral and cellular immune responses in the mice were then assessed using ELISA,flow cytometry,and ELISpot assays.Western blot analysis confirmed that the pp62 protein was successfully expressed,with a molecular weight of approximately 118.5 kDa.ELISA results indicated that a high level of specific antibodies was detected in the immunized mice,with antibody titers reaching up to 1∶1 638 400 at 7 days after the secondary immunization.The pro-portion of CD8+T lymphocytes in the immunized mice increased compared to the control group(P＜0.05).Results from the Q-PlexTM Mouse Cytokine Screen demonstrated that the secretion levels of IFN-γ,IL-2,IL-4,and IL-10 in serum were significantly upregulated in the immunized mice following secondary immunization(P＜0.001).In summary,these findings indicate that the pp62 protein can significantly stimulate both humoral and cellular immunity in mice,laying the groundwork for further studies on the function of the ASFV pp62 protein and the identification of novel vaccine antigens for ASF.

African swine fever(ASF)is a highly contagious disease caused by the African swine fe-ver virus(ASFV).To evaluate the immunogenicity of the pp62(CP530R)protein of ASFV,the recombinant CP530R protein was expressed in HEK 293F cells transfected with the plasmid pMAL-Fc-CP530R.Six-week-old female C57BL/6J mice were immunized with 10 μg of purified pp62 protein via subcutaneous injection,followed by a booster immunization with the same dosage at 21 days(enhanced).The humoral and cellular immune responses in the mice were then assessed using ELISA,flow cytometry,and ELISpot assays.Western blot analysis confirmed that the pp62 protein was successfully expressed,with a molecular weight of approximately 118.5 kDa.ELISA results indicated that a high level of specific antibodies was detected in the immunized mice,with antibody titers reaching up to 1∶1 638 400 at 7 days after the secondary immunization.The pro-portion of CD8+T lymphocytes in the immunized mice increased compared to the control group(P＜0.05).Results from the Q-PlexTM Mouse Cytokine Screen demonstrated that the secretion levels of IFN-γ,IL-2,IL-4,and IL-10 in serum were significantly upregulated in the immunized mice following secondary immunization(P＜0.001).In summary,these findings indicate that the pp62 protein can significantly stimulate both humoral and cellular immunity in mice,laying the groundwork for further studies on the function of the ASFV pp62 protein and the identification of novel vaccine antigens for ASF.

Objective To investigate the effects of simvastatin on the bone loss and deterioration of bone quality with different intervention programs. Methods Thirty two 3?month?old female Sprague?Dawley ( SD ) rats were randomized into 4 groups of 8 animals in each: All rats but those in the sham group ( A) received bilateral ovariectomy, and treated by vehicle (B), simvastatin (5 mg/kg/day) at first half period (C) or at latter half period (D). The rats in group C received simvastatin by a gavage after the OVX operation immediately, and stopped at 10 weeks after OVX. The rats in group D began to receive simvastatin treatment from 10 weeks after OVX and ended at 20 weeks after OVX. At week 20, all rats were sacrificed and the concentrations of serum PINP and ICTP were detected by ELISA, L3 vertabra was used to detect the bone mineral density, L4 vertebra was used to analyze the maximum loading and elastic modulus by compression test, and the microstructure of the L5 vertebra was detected by micro?computed tomography. Results 1. ELISA analysis:the concentrations of serum P1NP and ICTP in the groups A, B and C were significantly higher than that of group A (P<0?05). 2. BMD test showed that the rats in group B had significantly lower BMD than the other 3 groups (P<0?05), while the BMD of groups C and D were markedly lower than that of group A (P<0?05). 3. Biomechanical test:the maximum load and elastic modulus of L4 vertebrae in the group B were significantly lower than the other 3 groups ( P<0?05), and those of the groups C and D were markedly lower than that in the group A (P<0?05). 4. micro?CT:BV/TV and Tb. N in the sham operated rats were significantly higher than those of the other 3 groups, while the opposite trends was found in Tb. Sp (P<0?05), and the Tb. Sp in the groups C and D were significantly lower than that of group B (P<0?05). Conclusions Our data demonstrate that bone loss and deterioration of bone micro?structure and biomechanical properties occurred at 20 weeks after ovariectomy, both the first?half?period and latter?half?period treatment by simvastatin may partially prevent these changes, but can not restore the BMD to normal level.

Objective Alendronate is widely used as an anti-osteoporosis agent , while simvastatin , as a lipid-lowering drug , is being considered beneficial for bone formation .The present study aims to compare the effects of simvastatin and alendronate on bone loss in ovariectomized ( OVX) rats. Methods Thirty-two female SD rats were equally randomized into a sham operation , an OVX mod-el, a simvastatin ( OVX +S), and an alendronate ( OVX +A) group.All the rats but those in the sham operation group underwent dual ovariectomy .The animals of the OVX model and OVX +S groups were treated intragastrically with vehicle and simvastatin at 5 mg per kg of the body weight per day , respectively , while those of the OVX+A group injected subcutaneously with alendronate at 70μg per kg of the body weight per week .After 12 weeks of intervention , all the rats were sacrificed for detection of the serum concentrations of PINP and ICTP by ELISA, analysis of bone mineral density and bone histo-morphometry parameters of L 4 vertebrae , determination of the maximum loading and elastic modulus of L 5 vertebrae by compression test. Results The serum concentrations of P1NP and ICTP were significantly lower in the sham operation than in the other three groups (P<0.05), and that of ICTP markedly lower in the OVX +A than in the OVX model and OVX +S groups (P<0.05).Bone mineral density was remarkably lower in the OVX model ([0.238 ±0.007] g/cm2 ) than in the sham operation ([0.276 ±0.015] g/cm2), OVX+S ([0.250 ±0.014] g/cm2) and OVX+A groups ([0.269 ±0.014] g/cm2) (P<0.05), lower in the OVX+S than in the sham operation and OVX +A groups (P<0.05), but with no statistically significant difference between the latter two (P>0.05).Bone histomorphometry showed significantly lower BV /TV in the OVX model than in the sham operation and OVX +A groups ([19.9 ±1.69]%vs [29.03 ±2.59]%and [27.05 ±1.91]%, P<0.05), but markedly higher Tb.Sp in the former than in the latter two groups ([357.33 ±26.55] μm vs [211.17 ±16.56] μm and [252.50 ±19.35] μm, P<0.05).The maximum load and elastic modulus of L5 vertebrae were significantly lower in the OVX model rats than in the other three groups (P <0.05). Conclusion Both simvastatin and alendronate can inhibit bone loss in OVX rats , with comparable effects of preventing the deteriora-tion of biomechanical properties , but the latter is evidently more effective in maintaining bone mineral density .

Objective To investigate the prevalence of cognitive and motor disorders as well as emotional and sleep abnormality in the veterans from military communities in Beijing. Methods The participants underwent a comprehensive in-person evaluation including detailed neuropsychological testing,Hospital Anxiety and Depression Scale and special questionnaires for movement and sleep disorders. Results The overall prevalence of cognitive impairment, extrapyramidal diseases was 32.7%, 8.8% . The prevalence of mild cognitive impairment, dementia, Parkinson disease, essential tremor, anxiety and depression was 26.2% , 6.5% , 2.0% , 6.1 % , 1.4% and 4.1% respectively. Prevalence of all kinds of sleep disorders ranged from 10. 3% to 53. 9%. The prevalence of cognitive impairment had no significant difference of sex, but were correlated to age and education, the correlation coefficient was 0. 326 and -0.221 ( P<0.01) . Conclusion Veterans from military communities had higher prevalence of cognitive impairment, extrapyramidal diseases and sleep disorders and lower that of anxiety and depression relatively.