Objective:To explore the survival and prognostic factors of a long-course venetoclax-based(VEN-based)regimen in patients with de novo acute myeloid leukemia(AML)and provide evidence for the maintenance treatment of AML.Methods:A retrospective study was conducted in patients who received a VEN-based regimen and completed at least four courses of efficacy evaluation at The First Affiliated Hospital of Nanchang University from May 2021 to January 2024.The composite complete response rate(cCR),minimal residual disease(MRD)-negative rate,overall survival(OS)time,relapse-free survival(RFS)time,and adverse events were analyzed.Results:Overall,30 newly diagnosed patients with AML were enrolled in this study.The median age was 65(range,53-78)years,and the median number of treat-ment cycles was 7(range,4-20)years.After one cycle,the CR-and MRD-negative rates were 80.0%and 63.3%,respectively.The cumulative cCR was 96.7%,and MRD negative rate was 80.0%,respectively.The median follow-up time was 21.3(95%confidence intervals 14.7-27.9)months.The median OS time was 32.3 months and RFS time was not reached.The 2-year OS and RFS rates were 70.6%and 54.8%,respect-ively.Univariate analysis suggested that ELN2017 risk stratification and relapse status affected RFS and OS(P<0.05).However,the multivari-ate analysis failed to reveal any relationship between these factors and survival(P>0.05).In terms of safety,hematological adverse events were the most common,followed by infections.Overall,the VEN-based regimen was tolerated for patients with AML.Conclusions:A long-course VEN-based regimen is effective and safe.More than half of patients survive for>2 years,and it can be used as an effective mainten-ance treatment option for patients with AML.

Objective:To analyze the changes of DNA methylation in peripheral blood mononuclear cells (PBMC) of patients with hepatocellular carcinoma (HCC) and to evaluate the clinical value of a combined model based on FSIP1 gene methylation on the early diagnosis of HCC.Methods:This is a case-control study. From May 2023 to September 2024, 183 HCC patients and 155 healthy controls were collected in Qilu Hospital of Shandong University. The selected study subjects were divided into three cohorts: 14 HCC patients and 39 healthy controls formed the discovery cohort, a screening cohort consisted of 36 HCC patients and 39 healthy controls, 133 HCC patients and 77 healthy controls were included in the model construction cohort. 935k methylation chip analysis was used to identify specific differentially methylated sites in peripheral blood PBMC of the discovery cohort. The absolute value of the average methylation level difference between HCC group and healthy control group (|Δβ|) and P value were calculated. Then targeted bisulfite sequencing was used to verify the differentially methylated sites in the screening cohort. Finally, based on MethylTarget methylation sequencing technology, differential methylation sites were further verified in model construction cohort (divided into training set and validation set, training set consisted of 99 HCC patients and 57 healthy controls; validation set consisted of 34 HCC patients and 20 healthy controls). HCC early diagnosis model was constructed by random forest algorithm combined with clinical parameters and the diagnostic performance of the model was evaluated by receiver operating characteristic (ROC) curve in the validation set. Results:The total of 7 249 differentially methylated sites between HCC patients and healthy controls in discovery cohort were selected under the rule of |Δβ|≥0.06 and P<0.01. Among them, the cg02155073 site located on FSIP1 was hypermethylated in PBMC of HCC patients in the screening cohort and model cohort ( P<0.001). The AUC of HCC early diagnosis model (FmAP) based on FSIPI in the validation set was 0.967 (95% CI 0.924-1.000); sensitivity was 88%, specificity was 95%. The model had good diagnostic efficacy for patients with early HCC, stage Ⅰ-Ⅱ HCC AUC was 0.958 (95% CI 0.898-1.000). The FmAP model also had diagnostic value for tumor size <2 cm HCC and AFP negative HCC, with AUC of 0.955 (95% CI 0.898-1.000) and 0.964 (95% CI 0.934-0.994).The sensitivity were 92% and 93% and specificity both were 84%. Conclusion:The FmAP model based on FSIP1 gene methylation has good clinical value for the early diagnosis of hepatocellular carcinoma.

Objective:To explore the survival and prognostic factors of a long-course venetoclax-based(VEN-based)regimen in patients with de novo acute myeloid leukemia(AML)and provide evidence for the maintenance treatment of AML.Methods:A retrospective study was conducted in patients who received a VEN-based regimen and completed at least four courses of efficacy evaluation at The First Affiliated Hospital of Nanchang University from May 2021 to January 2024.The composite complete response rate(cCR),minimal residual disease(MRD)-negative rate,overall survival(OS)time,relapse-free survival(RFS)time,and adverse events were analyzed.Results:Overall,30 newly diagnosed patients with AML were enrolled in this study.The median age was 65(range,53-78)years,and the median number of treat-ment cycles was 7(range,4-20)years.After one cycle,the CR-and MRD-negative rates were 80.0%and 63.3%,respectively.The cumulative cCR was 96.7%,and MRD negative rate was 80.0%,respectively.The median follow-up time was 21.3(95%confidence intervals 14.7-27.9)months.The median OS time was 32.3 months and RFS time was not reached.The 2-year OS and RFS rates were 70.6%and 54.8%,respect-ively.Univariate analysis suggested that ELN2017 risk stratification and relapse status affected RFS and OS(P<0.05).However,the multivari-ate analysis failed to reveal any relationship between these factors and survival(P>0.05).In terms of safety,hematological adverse events were the most common,followed by infections.Overall,the VEN-based regimen was tolerated for patients with AML.Conclusions:A long-course VEN-based regimen is effective and safe.More than half of patients survive for>2 years,and it can be used as an effective mainten-ance treatment option for patients with AML.

Objective:To analyze the changes of DNA methylation in peripheral blood mononuclear cells (PBMC) of patients with hepatocellular carcinoma (HCC) and to evaluate the clinical value of a combined model based on FSIP1 gene methylation on the early diagnosis of HCC.Methods:This is a case-control study. From May 2023 to September 2024, 183 HCC patients and 155 healthy controls were collected in Qilu Hospital of Shandong University. The selected study subjects were divided into three cohorts: 14 HCC patients and 39 healthy controls formed the discovery cohort, a screening cohort consisted of 36 HCC patients and 39 healthy controls, 133 HCC patients and 77 healthy controls were included in the model construction cohort. 935k methylation chip analysis was used to identify specific differentially methylated sites in peripheral blood PBMC of the discovery cohort. The absolute value of the average methylation level difference between HCC group and healthy control group (|Δβ|) and P value were calculated. Then targeted bisulfite sequencing was used to verify the differentially methylated sites in the screening cohort. Finally, based on MethylTarget methylation sequencing technology, differential methylation sites were further verified in model construction cohort (divided into training set and validation set, training set consisted of 99 HCC patients and 57 healthy controls; validation set consisted of 34 HCC patients and 20 healthy controls). HCC early diagnosis model was constructed by random forest algorithm combined with clinical parameters and the diagnostic performance of the model was evaluated by receiver operating characteristic (ROC) curve in the validation set. Results:The total of 7 249 differentially methylated sites between HCC patients and healthy controls in discovery cohort were selected under the rule of |Δβ|≥0.06 and P<0.01. Among them, the cg02155073 site located on FSIP1 was hypermethylated in PBMC of HCC patients in the screening cohort and model cohort ( P<0.001). The AUC of HCC early diagnosis model (FmAP) based on FSIPI in the validation set was 0.967 (95% CI 0.924-1.000); sensitivity was 88%, specificity was 95%. The model had good diagnostic efficacy for patients with early HCC, stage Ⅰ-Ⅱ HCC AUC was 0.958 (95% CI 0.898-1.000). The FmAP model also had diagnostic value for tumor size <2 cm HCC and AFP negative HCC, with AUC of 0.955 (95% CI 0.898-1.000) and 0.964 (95% CI 0.934-0.994).The sensitivity were 92% and 93% and specificity both were 84%. Conclusion:The FmAP model based on FSIP1 gene methylation has good clinical value for the early diagnosis of hepatocellular carcinoma.

Objective:To screen the characteristic gut microbiota and fecal metabolites related to the efficacy of oxaliplatin-based neoadjuvant chemotherapy in patients with colorectal cancer liver metastasis, to analyze the relationship between gut microbiota and fecal metabolites, and to evaluate the predictive value of relevant markers for the efficacy of neoadjuvant chemotherapy in patients with colorectal cancer liver metastasis.Methods:This is a case-control study, 34 patients with colorectal cancer liver metastasis who were treated in Qilu Hospital of Shandong University from October 2021 to July 2022 were selected as the research objects, and were divided into chemotherapy effective group (20 cases) and chemotherapy ineffective group (14 cases) according to the efficacy evaluation criteria. Logistic regression was used to construct a prediction model to screen the microbiota and metabolic markers capable of predicting the effect of chemotherapy, and the receiver operating characteristic (ROC) curve and survival analysis curve were plotted to evaluate the predictive effect of related microbiota and metabolites on the efficacy of neoadjuvant chemotherapy.Results:There was no significant difference in the α and β diversity of gut microbiota between the patients in the chemotherapy effective group and in the ineffective group (all P>0.05). In terms of species, the relative abundance of 5 species was up-regulated and 10 species were down-regulated in the chemotherapy-effective group compared with the chemotherapy-ineffective group, and the difference was statistically significant (all P<0.05), among which Prevotella salivae could effectively predict the chemotherapy effect (AUC=0.750, P=0.007), with a sensitivity of 80.0% and a specificity of 71.4%. The overall survival of patients with high abundance (17 cases) was lower than that of patients with low abundance (17 cases) ( χ 2=5.239, P=0.022). In terms of metabolites, 20 metabolites were up-regulated and 4 metabolites were down-regulated in the chemotherapy-effective group compared with the chemotherapy-ineffective group, and the difference was statistically significant (all P<0.05), among which threonine and prostaglandin F2α-1-ethanolamide could distinguish between patients who responded to chemotherapy and those who did not respond to chemotherapy (AUC=0.743, 0.707, all P<0.05), and the overall survival of patients with high levels of relative abundance (17 cases) was higher than that of patients with low levels (17 cases) ( χ 2=4.748, 5.407, all P<0.05). The Logistic regression model of Prevotella salivae and prostaglandin F2α-1-ethanolamide was obtained through screening analysis, and the ROC curve results showed that the model had a good predictive value (AUC=0.836, sensitivity: 90.0%, specificity: 78.6%), and the overall survival of patients with high predict probability (17 cases) predicted by the model was higher than that of patients with low predict probability (17 cases) ( χ 2=9.260, P=0.002). Conclusion:Prevotella salivae and prostaglandin F2α-1-ethanolamide can be used as predictive biomarkers of neoadjuvant chemotherapy for colorectal cancer liver metastasis, and the model has good clinical reference value for prognosis assessment of patients in this cohort.

Objective:To explore the antibacterial performance and mechanism of reticulated cruciform DNA nanomachines mimicking neutrophil extracellular traps (NETs).Methods:In this study, a cross-shaped DNA nanostructure was synthesized using the one-step method to form a network under the polymerization of magnesium ions. The network was used as a template for inlaying reduced Cu 0 to form reticulated DNA-templated copper nanoclusters (CuNPs). The cruciform DNA formation process was characterized by polyacrylamide gel electrophoresis (PAGE). The reticulated cruciform DNA nanomachine was characterized by atomic force microscopy (AFM) and energy dispersive spectrometer (EDS). The optimal concentrations of Cu 2+, Mg 2+and DNA in the reticulated cruciform DNA nanomachine was determined by plate coating experiments. The antibacterial performance of the reticulated cruciform DNA nanomachine against Escherichia coli ( E. coli), Staphylococcus aureus and Klebsiella pneumoniae was verified by plate coating experiments. The aggregation effect of E. coli was evaluated by crystal violet staining, and the changes on the membrane surface of E. coli were observed by scanning electron microscopy (SEM), in order to explore the antibacterial mechanism of the reticulated cruciform DNA nanomachine. Results:PAGE showed that the band migration distance of the four DNA strands was the smallest after co-incubation. AFM showed that the DNA structure was reticulated and evenly distributed, and the Cu, Mg, and P elements coexisted in the structure. The optimal concentrations of Cu 2+, Mg 2+, and DNA required for the synthesis of the reticulated cruciform DNA nanomachine were 1 mmol/L, 100 mmol/L, and 20 μmol/L, respectively. The reticulated cruciform DNA nanomachine had the ability to inhibit E. coli, Staphylococcus aureus and Klebsiella pneumoniae, with survival rates of 39.72%, 43.56% and 60.57%, respectively. The reticulated cruciform DNA nanomachine had an aggregation effect on E. coli. The surface of the bacterial film exhibited shrinkage and fractures. Conclusion:The reticulated cruciform DNA nanomachine constructed in this study can aggregate E. coli, leading to the shrinkage and fracturing of the bacterial film, and exhibiting antibacterial effects.

Objective: : To investigate the efficacy and safety of the posterior endoscopic cervical foraminotomy (PECF) using ultrasonic osteotome for the treatment of cervical osseous foraminal stenosis，focusing on introduction of the advantages of ultrasonic osteotome in partial pediculectomy and ventral osteophyte resection in PECF. Methods: : Nineteen patients with cervical osseous foraminal stenosis who underwent PECF using ultrasonic osteotome in our institution between April 2018 and April 2021 were enrolled in this study. All the patients were followed up more than 12 months. The patients’ medical data, as well as pre- and postoperative radiologic findings were thoroughly investigated. The visual analogue score (VAS), Japanese Orthopaedic Association (JOA) score, cervical dysfunction index (Neck disability index, NDI), and modified MacNab criteria were used to assess the surgical efficacy. Results: : All the patients were successfully treated with PECF using ultrasonic osteotome. The pre- and postoperative VAS, NDI, and JOA scores were significantly improved (p<0.05). According to the modified MacNab criteria, 17 patients were assessed as “excellent”, two patients were assessed as “good” at the last follow-up. There was no dura tear, nerve root damage, incision infection, neck deformity, or other complications. Conclusion : Adequate nerve root decompression can be accomplished successfully with the help of ultrasonic osteotome in PECF, which has the advantage of reducing the probability of damage to the nerve root and dura mater, in addition to the original merits of endoscopic surgery.

Exosomes are nano-scale double-layer membrane structure vesicles that can be actively secreted by cells. They carry a large amount of biologically active substances and can serve as carriers for material transfer and information exchange between cells. In recent years, exosomes have become a frontier hotspot in biomedical research, and show broad application prospects in the field of laboratory diagnosis and clinical treatment of diseases. However, in general, most exosomes-related researchs are still in the laboratory research stage, and there are still many problems and challenges in separation and enrichment, technical operation specifications, and quality control. At the same time, it is urgent to carry out multi-center, large-sample clinical trials to provide evidence for exosomes from the laboratory to the clinical application.

Objective:To explore the effects of deleted in lung and esophageal cancer 1 (DLEC1) on osteosarcoma cells and the underlying mechanism.Methods:Immunohistochemical staining for DLEC1 was scored in sixteen paired osteosarcoma tissues and adjacent normal tissues obtained. The present study was conducted on human osteosarcoma 143B cells which were randomly divided into two groups, pDC316-DLEC1 transfection group and pDC316-Null transfection group. Differences in the proportion of EdU-positive cells, cell cycle distribution, proportion of apoptosis cells, number of migrating and invasive cells, expression of epithelial-mesenchymal transformation (EMT) markers (E-cadherin and vimentin), relative protein expression levels of NF-κB, AKT and ERK signaling pathways were assessed between the pDC316-DLEC1 and pDC316-Null transfection groups in in vitro study. The subcutaneous inoculation model and tail vein injection model were developed to evaluate the differences in subcutaneous tumor volume, subcutaneous tumor weight and pulmonary tumor nodules between the above two groups in in vivo study.Results:The DLEC1 immunostaining scores for osteosarcoma tissues and adjacent normal tissues were 2.88±1.15 and 4.25±1.06, respectively. The proportions of EdU-positive cells (36.47%±1.90% vs 51.47%±2.89%) and S phase cells (33.31%±0.61 vs 43.77%±1.47%) were decreased, while G0/G1 phase cells (46.87%±0.73% vs 35.47%±1.14%) and apoptotic cells (13.83%±1.01% vs 3.30%±0.26%) were increased in the pDC316-DLEC1 transfection group compared to those in the pDC316-Null transfection group. Decreased number of migrating cells (199.00±12.53 vs 369.67±10.02) and invasive cells (104.67±9.07 vs 299.67±12.06) and relative expression of vimentin mRNA (0.59±0.02 vs 1.00±0.02) and protein (0.54±0.08 vs 1.00±0.00) were observed in the pDC316-DLEC1 transfection group, while relative expression of E-cadherin mRNA (2.40±0.05 vs 1.00±0.02) and protein(1.98±0.10 vs 1.00±0.00) in the pDC316-DLEC1 transfection group were higher than those in the pDC316-Null transfection group. The relative protein expression of NF-κB (p65), p-AKT (Ser473) and p-ERK (Thr202/Tyr204) in the pDC316-DLEC1 transfection group were decreased by 51.67%±4.04%, 64.67%±5.51% and 48.67%±4.73% compared to those in the pDC316-Null transfection group. In in vivo study, 143B cells in the pDC316-DLEC1 transfection group formed smaller (320.00±145.22 mm 3vs 798.00±221.94 mm 3) and lighter (0.49±0.17 g vs 0.88±0.14 g) subcutaneous tumors and less metastatic lung nodules (7.71±1.80 vs 20.86±3.53) compared with those in the pDC316-Null transfection group. Conclusion:Overexpression of DLEC1 could suppress the NF-κB/AKT/ERK signaling pathways in 143B cells, which further induces G0/G1 arrest and apoptosis that ultimately inhibits cell proliferation and reduces the metastatic potential through reversing EMT.

Objective Next Generation Sequencing(NGS) platform was used to study the characteristics of hot gene mutations in non-small cell lung cancer (NSCLC). The distribution, type and frequency of mutation sites were systematically analyzed to evaluate the pathogenicity of mutation sites . Methods A total of 94 NSCLC tissue samples were included in this study including paraffin-embedded (FFPE) samples and fresh tissue samples, which were collected from July 2015 to April 2017 at the Qilu Hospital of Shandong University. The patient's age ranged from 35 to 82 years with a median age of 61 years. There were 63 males and 31 females. 22 hot genes in NSCLC were selected as the detection panel, including KRAS, EGFR, BRAF, PIK3CA, AKT1, ERBB2, PTEN, NRAS, STK11, MAP2K1, ALK, DDR2, CTNNB1, MET, TP53, SMAD4, FBXW7, NOTCH1, ERBB4, FGFR1 and FGFR2. Mutation detection was performed using the Ion AmpliSeq Colon and Lung Cancer Panel of the Thermo fisher's Ion Torrent sequencing platform. The sequencing data was analyzed using Ion Torrent suite v4.4.2 software. Results Among the 22 mutant genes commonly found in NSCLC, the mutation frequency of TP53 was the highest, accounting for 46.9％ of all mutations, followed by the EGFR mutation (28.1％); A total of 89 mutations were detected, including 63 hot spot mutations (reported mutations) and 26 new mutations (unreported mutations). The most frequently detected mutation was the frameshift deletion of exon 19 of EGFR, followed by the mutation of exon L858R;Analysis of the mutation in targeted drug sites of EGFR showed that the frameshift deletion of exon 19 of EGFR was the most frequently detected, followed by the mutation of exon L858R on chromosome 21. Bioinformatics software was used to analyze the pathogenicity of 26 new mutation sites. Results showed that in addition to ATK1:c. 47-12G>A and TP53: c. 214 C>G, the remaining 24 new mutation sites had at least one major impact on the gene function in three aspects, including gene conservation, amino acid sequence change and protein structure influence. Conclusion In this study, NGS was used to conduct combined detection of mutation sites of multiple hot genes, which might cover more comprehensively genetic variation and provide a basis for screening the most suitable targeted therapy groups. The pathogenicity prediction of new mutations and the changes in tumor-related signaling pathways involved provide a reference for further study of the pathogenesis of NSCLC.