ABSTRACT
Emerging evidence has pointed out the importance of long non-coding RNAs (lncRNAs) in multiple diseases, the knowledge of rheumatoid arthritis (RA)-associated lncRNAs remains limited. In this present study, we aimed to elucidate the mechanism of lncRNA nuclear paraspeckle assembly transcript 1 (NEAT1) from peripheral blood monouclear cell (PBMC)-derived exosomes (exos) on RA development by modulating the microRNA-23a (miR-23a)/murine double minute-2 (MDM2)/Sirtuin 6 (SIRT6) axis. RA was modeled in vivo by collagen induction in mice and in vitro by exposing fibroblast-like synoviocytes (FLSs) to lipopolysaccharide. Exos were isolated from human or mouse PBMCs, which were then were co-cultured with FLSs. Based on gain- and loss-of-function experiments, the cell proliferation and secretion of inflammatory factors were measured. LncRNA NEAT1 was found to be highly expressed in RA, and PBMCs-derived exos contributed to RA development by delivering lncRNA NEAT1. In lipopolysaccharide-induced FLSs, miR-23a inhibited the expression of MDM2, and overexpression of MDM2 partially rescued the inhibitory effect of miR-23a on FLS proliferation and inflammatory response. Mechanistically, MDM2 ubiquitination degraded SIRT6 in RA. LncRNA NEAT1 shuttled by PBMC-derived exos promoted FLS proliferation and inflammation through regulating the MDM2/SIRT6 axis. Furthermore, in vivo experiments suggested that downregulated lncRNA NEAT1 shuttled by PBMC-derived exos or upregulated miR-23a impeded RA deterioration in mice. This study highlights that lncRNA NEAT1 shuttled by PBMC-derived exos contributes to RA development with the involvement of the miR-23a/MDM2/SIRT6 axis.
ABSTRACT
Objective To study the effects of tigecycline on immune function in patients with complicated intra-abdominal infections (cIAI). Methods A total of 24 cIAI patients received treatment of tigecycline, and then the effects of tigecycline on level of peripheral blood monouclear cells (PBMC) proliferation, concentration of inflammatory cytokines, and expression of CD3, CD4 and CD8 were investigated. Results The total effective rates of tigecycline were 70. 8％. Tigecycline treatment significantly reduced proliferative level of PBMC, decreased the levels of interleukin-1β (IL-1β), IL-6 and IL-8 in supernatants of PBMC cultures as well as serum. Moreover, tigecycline therapy significantly up-regulated the percentage of CD3 + and CD4 +, increased the ratio of CD4+ / CD8 +, and down-regulated percentage of CD8 + in peripheral blood. Conclusion Tigecycline could regulate immune function in cIAI patients.
ABSTRACT
This study aimed to investigate the effects of vascular endothelial growth factor (VEGF) secreted by MCF-7 breast cancer cells on the differentiation, maturation and function of dendritic cells (DCs). Small interfering RNAs (siRNAs) directed against the VEGF gene were designed and transfected into MCF-7 breast cancer cells at an optimal concentration (100 nmol/l) using cationic liposome transfection reagent, whereas the control group was transfected with only transfection reagent. Western blot analysis and ELISA were used to determine VEGF protein expression and VEGF concentration, respectively. Mononuclear cells were cultured with the culture supernatants from primary MCF-7 cells (control group) and siRNA-treated MCF-7 cells (siRNA group). The DC phenotypes, including CD1a, CD80, CD83, CD86 and HLA-DR, were evaluated by flow cytometry. The MTT assay was used to assess the cytotoxicity of DC-mediated tumor-specific cytotoxic T lymphocytes (CTLs) against MCF-7 cells in the two different culture supernatants. The VEGF-targeted constructed siRNA inhibited VEGF expression in MCF-7 cells. Cultivation with the culture supernatants from MCF-7 cells treated with siRNA affected DC morphology. DCs in the siRNA group exhibited a significantly higher expression of CD86, CD80, CD83 and HLA-DR compared to the cells in the control group, whereas the expression of CD1a in the siRNA group was significantly lower compared to that in the control group. The cytotoxic activity of CTLs mediated by DCs was significantly altered by siRNA transfection. These results indicated that VEGF may play a significant role in tumor development, progression and immunosuppression.
ABSTRACT
OBJECTIVE: To identify the characteristics of recombinant adenovirus modified PBMC-derived dendritic cells to resist the HIV-1 infection. METHODS: The recombinant adenovirus vector was integrated with CCR5A32, CCR5siRNA, HIV-1 poli and HIV-1 inti genes by using AdEasy system. The human PBMC from healthy donor blood was isolated in order to get dendritic cells. The expression of CCR5Δ32, CCR5, CXCR4 and HIV-1 P24 in PBMC or modified cells was measured by Westernblot. RESULTS: After the cells have been modified by Ad-R5Δ32siHIV, the expression of CCR5Δ32 was increased while the expression of CCR5 and CXCR4 were decreased. The level of p24 was lower in modified cells than that of unmodified cells. The modified cells show resistance to HIV-1 infection. CONCLUSION: The recombinant adenovirus-modified cells show good resistance to HIV-1 infection. Modification of HSC-derived immunity cells, such as DCs, may be a potent strategy to resist HIV-1 infection.
ABSTRACT
OBJECTIVE: To study whether the therapeutic action of compound glycyrrhizin(SNMC) on allergic disorders can be achieved through inhibition on the expression of chemotactic factors in peripheral blood mononuclear cells(PBMC).METHODS: PBMC of 15 healthy subjects were obtained and divided into 4 drug-treated groups: SNMC group,dexamethasone group,mizolastine group,and loratadine group;meanwhile,a positive control group(proinflammatory stimulant was added) and a negative control group(no proinflammatory stimulant was added) were used as control.Fluorescent quantitation polymerase chain reaction was employed to determine the mRNA expression of the chemotatic factors including RANTES,MCP-1,MIP-1? and ELISA was employed to detect their concentrations in supernatant.RESULTS: In the positive control group,both the mRNA expressions and the concentrations of the 3 chemotatic factors increased significantly,and which were significantly lower in the drug-treatment groups than in the positive control group.There were significant differences between the positive control group and dexamethasone-treated group or SNMC-treated group in the expression of MIP-1? mRNA(P
