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Safe and effective vaccines are the best method to defeat worldwide SARS-CoV-2 and its circulating variants. The SARS-CoV-2 S protein and its subunits are the most attractive targets for the development of protein-based vaccines. In this study, we evaluated three lipophilic adjuvants, monophosphoryl lipid A (MPLA), Toll-like receptor (TLR) 1/2 ligand Pam

Adjuvants are important components in vaccines to increase the immunogenicity of proteins and induce optimal immunity. In this study, we designed a novel ternary adjuvant system Alum + c-GAMP + poly(I:C) with STING agonist 3,3'-c-GAMP (c-GAMP) and TLR3 agonist poly(I:C) co-adsorbed on the conventional adjuvant aluminum gel (Alum), and further constructed an S1 protein vaccine. Two doses of vaccination with the ternary adjuvant vaccine were sufficient to induce a balanced Th1/Th2 immune response and robust humoral and cellular immunity. Additionally, the ternary adjuvant group had effective neutralizing activity against live virus SARS-CoV-2 and pseudovirus of all variants of concern (alpha, beta, gamma, delta and omicron). These results indicate that the ternary adjuvants have a significant synergistic effect and can rapidly trigger potent immune responses; the combination of the ternary adjuvant system with S1 protein is a promising COVID-19 vaccine candidate.

This study aims to detect the biological effects of matrix metalloproteinase 3 (MMP3) and SRY-related high mobility group-box gene 9 (SOX9) gene regulation in the human intervertebral disc degeneration of nucleus pulposus cells mediated by lentiviral vectors in vitro. The culture and development of human degenerated intervertebral disc nucleus pulposus cells was performed. The cell morphology and structure were observed, as identified by hematoxylin and eosin (H&E) staining. The experiment was divided into five groups: blank control group, SOX9 gene overexpression group, MMP3 gene silencing group, SOX9 gene overexpression+MMP3 gene silencing group, and blank vector group. RT-PCR and western blot were performed to detect the influence of MMP3 gene silencing and SOX9 gene overexpression on degenerated human intervertebral disc nucleus pulposus cells, include the secretion of polysaccharide and expression of type II collagen, at the mRNA and protein level. The originally generated degenerated human intervertebral disc nucleus pulposus cells had the same cellular morphology. The MTT assay revealed that the blank group and blank vector group had no statistical significance in cell proliferation. The MMP3 gene silencing and SOX9 gene overexpression could promote the proliferation of degenerated human intervertebral disc nucleus pulposus cells, when compared with the blank control group and blank vector group, and this more obvious in the MMP3 gene silencing group and SOX9 gene overexpression group. RT-PCR and western blot revealed that MMP3 gene silencing and SOX9 gene overexpression can promote cells to secrete polysaccharide and type-II collagen (P < 0.05). MMP3 gene silencing and SOX9 gene overexpression can promote the proliferation of degenerated human intervertebral disc nucleus pulposus cells, and their ability to secrete polysaccharide and type-II collagen.

Enzyme-labeled secondary antibody is often used to amplify the output signal in the process of antibody detection. However, its preparation process is complex and time-consuming. Herein, we fabricated an innovative hydrophilic rhodamine B-loaded / boronic acid-modified graphene oxide (HRBGO) nanocomposite, used as a substitute of enzyme-labeled second antibody. The synthetic HRBGO was loaded with generous rhodamine B and modified with boronic acid. Therefore, the HRBGO could selectively label the carbohydrate chains of Fc fragment of primary antibody through specific boronate affinity recognition, and then perform signal output and amplification by releasing rhodamine B. To verify the practicability of HRBGO, trastuzumab as a humanized monoclonal antibody targeting human epidermal growth factor receptor-2 (HER2) was selected as model antibody. A glycosylation site-blocked / HER2-immobilized magnetic nanoparticles (GHMN) was also prepared for selectively capturing trastuzumab from complex samples via specific immunoaffinity. Because the glycosylation sites of HER2 can also be labeled with the HRBGO by boronate affinity recognition, these sites were blocked by a masking agent to minimize the background signal. For specific and ultrasensitive detection of trastuzumab, the integration of GHMN and HRBGO was proposed and optimized in detail. Trastuzumab detection based on HRBGO consisted of three steps: specific capture, selective labeling, and output signal. The proposed strategy provided ultrahigh sensitivity with limit of detection of 0.35 fg mL

The quality of the base layer materials is one of the key factors that influences pavement performance and service life. The previous research and experiment results have preliminarily proved that cool regenerated materials have good road-applied properties, and verified the main technical characteristics of preliminary analysis results of cement cold regenerated. Based on experimental results and cement strength standard, road performance cold regenerated aggregates and cold regenerated lime treated soil is analyzed in this paper. The results show that the performance of cement cold regenerated materials meets the requirements. Cement cold regenerated aggregates can be used as Grade 2nd and below Grade 2nd road. Under appropriate conditions, cement cold regenerated aggregates can also be used for freeway base; Cement cold regenerated lime treated soil can be used as subbase Grade 2nd and Grade 2nd below road.

In order to study the factors which influence the inorganic binder stabilized material’s compaction test results, different recycled material content therefore different reclaimed gradation mixtures are tested while varying the amount of cement. The experiment results show that contents of the recycled base material and the recycled surface material on the compaction test results are determined by the change of their density and water absorption ratio. Dry density of the recycled mixture increases with the increase of the reclaimed base material density. Low water absorption ratio of the recycled material reduces the reclaimed mixture’s optimum moisture content. Density of the reclaimed wearing surface material reduces the maximum dry density of the reclaimed mixture because the old wearing surface material has lower density. Its low water absorption reduces the optimum moisture content of the recycled mixture. Influence of cement content on compaction test results is the increase of the cement content can enhance the maximum dry density and optimum moisture content of the recycled material. Through the analysis of the results of the compaction test, the key factors in the recycled material compaction test are unveiled.