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.

To explore the feasibility and accuracy for evaluating mitral regurgitation (MR) severity with MR jet volume (MRvol) by means of general imaging three-dimensional quantification (GI3DQ). Methods: A total of 93 MR patients were divided into 2 groups: Central MR group, n=41 and Eccentric MR group, n=52. According to real-time three-dimensional echocardiography (RT3DE) examined planimetry of effective regurgitation orifice area (EROA), the patients were graded into mild MR, moderate MR and severe MR. MRvol was directly measured by GI3DQ. Results: In Central MR group, ROC analysis showed that as GI3DQ measured MRvol>16.2 ml, AUC=0.93, P<0.0001, the sensitivity and specificity for distinguishing mild MR and moderate MR were 96.0% and 70.0%respectively; as MRvol>44.5 ml, AUC=0.96, P<0.0001, the sensitivity and specificity for distinguishing moderate MR and severe MR were 97.6% and 91.7% respectively. In Eccentric MR group, as MRvol>14.2 ml, AUC=0.77, P=0.0243, the sensitivity and specificity for differentiating mild MR and moderate MR were 91.8% and 62.5% respectively; as MRvol>40.5 ml, AUC=0.83, P<0.0001, the sensitivity and specificity for differentiating moderate MR and severe MR were 82.3% and 77.9% respectively. Conclusion: Taking RT3DE examined EROA as reference, GI3DQ directly measured MRvol could more accurately assess MR severity especially in patients with central MR, it may distinguish moderate MR and severe MR with the higher sensitivity and specificity.

Objective To evaluate diagnostic value of mitral regurgitation jet volume(MRvol) in the quantification of mitral regurgitation severity by general imaging three-dimensional quantification (GI3DQ) using the guideline recommended 2D integrative method as a reference.Methods Ninety-three patients with MR were divided into central MR group(n =41) and eccentric MR group(n =52).The American Society of Echocardiography (ASE)-recommended 2D integrative method was used as a reference for MR grading and MRvol was directly measured by GI3DQ method.Results In central MR,as assessed by receiver operating characteristic (ROC) analysis,the area under the curve(AUC)was 0.87(P <0.0001), and MRvol by GI3DQ at a cutoff value of 16.2 ml yielded 96.2% of sensitivity and 63.6% of specificity to differentiate mild from moderate MR;the AUC was 0.98(P < 0.0001),and a cutoff value of 47.8 ml yielded 98.6% of sensitivity and 96.2% of specificity to differentiate moderate from severe MR. In eccentric MR,the AUC was 0.76(P =0.086),and MRvol at a cutoff value of 14.8 ml yielded 90.9% of sensitivity and 60.0% of specificity to differentiate mild from moderate MR;the AUC was 0.84(P <0.0001) and a cutoff value of 40.7 ml yielded 80.0% of sensitivity and 79.7% of specificity to differentiate moderate from severe MR.Conclusions MRvol measured directly by GI3DQ could more exactly evaluate MR severity,and have better sensitivity and specificity to differentiate moderate from severe MR in central MR.