Objective To investigate the molecular mechanism of hepatic fibrosis(HF)regulation by liver sinusoidal endothelial cells(LSECs)via endothelial mesenchymal transition(EnMT),and to predict the natural active components using bioinformatics,machine learning,and cellular experiments.Methods HF and EnMT gene matrices were obtained and the intersecting genes were extracted and enriched using Limma difference analysis and weighted gene co-expression network analysis(WGCNA).The diagnostic genes were screened using a combination of random forest method,support vector machine-recursive feature elimination and network topology analysis,and immune infiltration analysis and prediction of natural active ingredients were performed.The expression of diagnostic genes and the pharmacological effects of the predicted ingredients were finally verified by cellular experiments.Results Differential analysis yielded 3034 EnMT-associated and 4133 HF-associated differential genes.WGCNA analysis yielded 4589 EnMT-associated Hub genes and 763 HF-associated Hub genes.Thirty-eight intersecting genes were extracted,which were mainly enriched in the pathways of basement membrane and extracellular matrix receptor interaction.Four diagnostic genes,CFP,COL4A2,ITGA1,and GRPEL1,were screened by multidimensional analysis.Immune infiltration analysis showed that the diagnostic genes were closely associated with mast cell resting state,memory B cells,and memory CD4+T cells.Reverse transcription-polymerase chain reaction analysis showed significantly increased mRNA expression levels of the four diagnostic genes in the Jagged1-induced model group(P＜0.05).The predicted components,sterol,kaempferol,and quercetin,all had good binding activities with the diagnostic genes.Enzyme-linked immunosorbent assay result confirmed that all three active components significantly reduced the expression of collagen type Ⅳ α2 chain protein in Jagged1-induced LSECs,with quercetin having the most significant effect(P＜0.01).Conclusions This study elucidated the molecular mechanism of hepatic sinusoidal endothelial cells involved in the pathological process of HF through mesenchymal transition.We also propose a diagnostic marker system including CFP,COL4A2,ITGA1,and GRPEL1 as core genes.The result also suggest that natural active ingredients,such as quercetin,may exert anti-HF pharmacological effects by targeting these diagnostic genes.

Objective To investigate the molecular mechanism of hepatic fibrosis(HF)regulation by liver sinusoidal endothelial cells(LSECs)via endothelial mesenchymal transition(EnMT),and to predict the natural active components using bioinformatics,machine learning,and cellular experiments.Methods HF and EnMT gene matrices were obtained and the intersecting genes were extracted and enriched using Limma difference analysis and weighted gene co-expression network analysis(WGCNA).The diagnostic genes were screened using a combination of random forest method,support vector machine-recursive feature elimination and network topology analysis,and immune infiltration analysis and prediction of natural active ingredients were performed.The expression of diagnostic genes and the pharmacological effects of the predicted ingredients were finally verified by cellular experiments.Results Differential analysis yielded 3034 EnMT-associated and 4133 HF-associated differential genes.WGCNA analysis yielded 4589 EnMT-associated Hub genes and 763 HF-associated Hub genes.Thirty-eight intersecting genes were extracted,which were mainly enriched in the pathways of basement membrane and extracellular matrix receptor interaction.Four diagnostic genes,CFP,COL4A2,ITGA1,and GRPEL1,were screened by multidimensional analysis.Immune infiltration analysis showed that the diagnostic genes were closely associated with mast cell resting state,memory B cells,and memory CD4+T cells.Reverse transcription-polymerase chain reaction analysis showed significantly increased mRNA expression levels of the four diagnostic genes in the Jagged1-induced model group(P＜0.05).The predicted components,sterol,kaempferol,and quercetin,all had good binding activities with the diagnostic genes.Enzyme-linked immunosorbent assay result confirmed that all three active components significantly reduced the expression of collagen type Ⅳ α2 chain protein in Jagged1-induced LSECs,with quercetin having the most significant effect(P＜0.01).Conclusions This study elucidated the molecular mechanism of hepatic sinusoidal endothelial cells involved in the pathological process of HF through mesenchymal transition.We also propose a diagnostic marker system including CFP,COL4A2,ITGA1,and GRPEL1 as core genes.The result also suggest that natural active ingredients,such as quercetin,may exert anti-HF pharmacological effects by targeting these diagnostic genes.

Objective To investigate the predictive and guiding significance of peripheral blood biomarkers on the therapeutic effects of PD-1/PD-L1 inhibitor treatment on lung cancer patients. Methods We collected the data of 200 lung cancer patients treated with PD-1/PD-L1 inhibitors treatment, including clinical indicators, peripheral blood indicators, efficacy indicators and survival indicators. Results The DCR of patients with non-hepatic metastasis, immune combined chemotherapy, NLR≤2.81 and LDH≤202.5 u/L was higher (P < 0.05). The AUC value of NLR combined with LDH predicting DCR was 0.698 (P < 0.05). Univariate analysis showed that non-hepatic metastasis, first-line immunotherapy, immunotherapy combined with chemotherapy and LDH≤202.5 u/L were related to PFS (P < 0.05). Multivariate analysis showed that the patients with non-hepatic metastasis and LDH≤202.5 u/L had longer PFS (P < 0.05). The significant decrease of NLR and LDH after two cycles of immunotherapy indicated the effectiveness of immunotherapy (P < 0.05). Conclusion NLR≤2.81, LDH≤202.5 u/L, non-hepatic metastasis and immunotherapy combined chemotherapy are positively correlated with immunotherapy efficacy. Non-hepatic metastasis and LDH≤202.5 u/L are independent prognostic factors of the patients treated with immunotherapy. The changes of peripheral blood NLR and LDH are related to the efficacy of PD-1/PD-L1 inhibitors treatment.