Immunotherapy has emerged as a revolutionary approach to treat immune-related diseases. Dendritic cells (DCs) play a pivotal role in orchestrating immune responses, making them an attractive target for immunotherapeutic interventions. Modulation of gene expression in DCs using genome editing techniques, such as the CRISPR-Cas system, is important for regulating DC functions. However, the precise delivery of CRISPR-based therapies to DCs has posed a significant challenge. While lipid nanoparticles (LNPs) have been extensively studied for gene editing in tumor cells, their potential application in DCs has remained relatively unexplored. This study investigates the important role of cholesterol in regulating the efficiency of BAMEA-O16B lipid-assisted nanoparticles (BLANs) as carriers of CRISPR/Cas9 for gene editing in DCs. Remarkably, BLANs with low cholesterol density exhibit exceptional mRNA uptake, improved endosomal escape, and efficient single-guide RNA release capabilities. Administration of BLAN_mCas9/gPD-L1 results in substantial PD-L1 gene knockout in conventional dendritic cells (cDCs), accompanied by heightened cDC1 activation, T cell stimulation, and significant suppression of tumor growth. The study underscores the pivotal role of cholesterol density within LNPs, revealing potent influence on gene editing efficacy within DCs. This strategy holds immense promise for the field of cancer immunotherapy, offering a novel avenue for treating immune-related diseases.

Objective To explore the application effect of sandwich teaching method in clini-cal teaching in the department of orthopedics. Methods Totally 150 clinical medicine students were divided into 2 groups: sandwich teaching group (n=75) and lecture-based learning group (n=75). Sandwich teaching method combined with bedside teaching method was used in sandwich teaching group while lecturing combined with bedside teaching method was used in control group. Theoretical examination, clinical operation skill test, clinical case analysis and questionnaire investigation were conducted after the course. Comparison between the two groups was made using independent sample t test and measurement data were expressed as x±s. P<0.05 signifies statistically significant differences. Results Theoretical examination score, clinical skill test score and clinical case analysis score were higher in sandwich teaching group than in control group. [(53.70±4.27) vs. (48.00±4.83);(15.70± 2.49) vs. (11.40±2.87);(17.10±1.52) vs. (13.80±1.32)]. Questionnaire showed that commutation and expression ability, cooperation ability and self thinking ability were better in sandwich teaching group than in control group , with statistical differences . Conclusions Sandwich teaching method achieves good teaching effect and it is worth recommending in clinical teaching.