Objective To investigate the gene expression and regulatory mechanisms of mouse embryonic fibroblasts (MEFs) under inflammatory conditions, aiming to elucidate the role of MEFs in inflammatory responses and provide a foundation for discovering anti-inflammatory drugs that act by modulating MEF function. Methods MEFs cultured in vitro were divided into the following groups: lipopolysaccharides (LPS)-treated group, inflammatory conditioned medium (CM)-treated group, and control group, which were treated with LPS, CM, and equal volume solvent, respectively. Transcriptome sequencing was used to analyze the effects of two stimuli on gene expression profile of MEFs. Real time fluorescence quantitative PCR (RT-qPCR) was employed to verify the transcription levels of highly expressed genes of MEFs induced by CM. ELISA was performed to determine the concentrations of cytokines in cell supernatants. Finally, the regulatory effects of CM on the activation of signaling pathways in MEFs were analyzed by immunoblotting. Results Transcriptome analysis showed that both LPS and CM induced the transcription of a large number of genes in MEFs. Compared with LPS, CM potentiated the mRNA transcription of some acute phase proteins, inflammatory cytokines, chemokines, matrix metalloproteinases (MMP), prostaglandin synthetases, and colony-stimulating factors. The transcriptome analysis was verified by RT-qPCR. The results of ELISA showed that CM treatment significantly increased the secretion of interleukin 6 (IL-6), C-C motif chemokine ligand (CCL2), and C-X-C motif chemokine ligand (CXCL1) by MEFs compared with LPS. Mechanism study showed that both LPS and CM induced the phosphorylation of nuclear factor-κB p65 (NF-κB p65), p38 mitogen-activated protein kinase (p38 MAPK), extracellular regulated protein kinases 1/2 (ERK1/2), and TANK-binding kinase (TBK) in MEFs, and CM strongly stimulated the phosphorylation of signal transducer and activator of transcription 3 (STAT3) in MEFs. Conclusion Both LPS and CM can induce transcription and protein secretion of various inflammation-related genes in MEFs. CM can partly enhance LPS-induced activation of MEFs, and the mechanism may be related to the enhancement effect of CM on the activation STAT3 signaling pathway.
Animals ; Fibroblasts/immunology* ; Mice ; Lipopolysaccharides/pharmacology* ; Inflammation/metabolism* ; Signal Transduction/drug effects* ; Gene Expression Regulation/drug effects* ; Cytokines/genetics* ; Culture Media, Conditioned/pharmacology* ; Cells, Cultured

OBJECTIVE To optimize hematopoietic stem cell transplantation therapy and provide support for drug research by investigating the dynamic process of hematopoietic and immune system reconstitution after bone marrow transplantation(BMT)in mice.METHODS CD45.2+C57BL/6 mice were used as recipient mice and randomly divided into the normal control group and transplantation group,with 30 mice in each.The transplantation group was irradiated by a lethal dose of cobalt-60 rays.Bone marrow cells were prepared from CD45.1+C57BL/6 mice and transfused into recipient mice through the tail vein.Peripheral blood,spleens,lymph nodes,thymuses and bone marrow were collected at 1,2,4,8 and 16 weeks after transplantation.Blood routine examination was performed with peripheral blood and total cell numbers in suspensions of other organs were counted by an automated cell counter.Cell classification analysis of white blood cells in peripheral blood,cell suspensions of other organs was performed by flow cytometry.RESULTS Four weeks after BMT,the numbers of white blood cells and red blood cells in peripheral blood of recipient mice returned to the same level of or higher level than normal control(P<0.05).Although the number of platelets recovered significantly,it was still mark-edly lower than that of normal control until 16 weeks post BMT(P<0.05).In addition,the percentages of myeloid leukocytes and B cells in peripheral blood,spleens,lymph nodes,and bone marrow,as well as megakaryocytes and erythrocyte progenitor cells in bone marrow also returned to normal,and the majority of myeloid leukocytes and B cells were CD45.1+cells from the donors.Eight weeks after BMT,T cells in peripheral blood,spleens,lymph nodes,thymuses,and bone marrow of recipient mice returned to normal,and CD45.1+T cells were dominating.CONCLUSION The hematopoietic and immune reconstitution of recipient mice is nearly completed eight weeks after BMT.However,the reconstruction speed of different kinds of cells and the reconstruction status of same kind of cell in different organs vary widely.

Objective To summarize and analyze the experience of nursing patients with congenital heart disease and pulmonary hypertension undergoing intervention therapy.Method The nursing measures included psychological nursing, preoperative preparation,postoperative monitoring of pulmonary hypertension and vital signs, observation and prevention of complications,and transfusion management.Result All the 18 patients recovered well after intervention therapy without severe complications.Conclusion The peri-intervention nursing of patients with congenital heart disease and pulmonary hypertension is important for increasing interventional success rate and lowering postoperative complication.