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.

Deep proximal lesions with significant defects and subgingival margins exceeding cementoenamel junction are common clin-ical scenarios.With the development of materials and bonding dentistry,along with the popularization of the concept of minimally inva-sive dentistry,deep margin elevation is becoming an option for its minimal invasion and less time-consuming.Even though deep margin elevation seems a valuable technique,clinicians have not extensively applied it limited to its technical sensitivity.Current research is mainly in vitro and lacks high-quality clinical randomized controlled trails,or even standardized clinical process.This review summari-zes the concept,procedure and clinical performance of deep margin elevation.

Deep proximal lesions with significant defects and subgingival margins exceeding cementoenamel junction are common clin-ical scenarios.With the development of materials and bonding dentistry,along with the popularization of the concept of minimally inva-sive dentistry,deep margin elevation is becoming an option for its minimal invasion and less time-consuming.Even though deep margin elevation seems a valuable technique,clinicians have not extensively applied it limited to its technical sensitivity.Current research is mainly in vitro and lacks high-quality clinical randomized controlled trails,or even standardized clinical process.This review summari-zes the concept,procedure and clinical performance of deep margin elevation.

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 understand the epidemiological characteristics and post-exposure prophylaxis (PEP) situation of rabies exposure population in the animal injury outpatient department of Lishui Central Hospital, so as to provide reference for rabies prevention and control in this region.Methods:The epidemiological data of 6 471 rabies-exposed persons in Lishui Central Hospital from 2021 to 2023 were retrospectively analyzed, including the gender and age of exposed persons, the month and location of injury, the species of injured animals, the exposure grade, rabies vaccination, rabies passive immunization agents, etc.Results:From 2021 to 2023, a total of 6 471 cases of rabies exposure were treated in the animal injury outpatient department of Lishui Central Hospital. From 2021 to 2023, 1 133 cases, 2 135 cases and 3 203 cases were treated respectively. April to November was the peak period of exposure. The population of 21 to 30 years had the highest rate of treatment, reaching 27.79% (1 798/6 471), and was a high-risk group of exposure. The age composition ratio of rabies exposure in the three years was statistically significantly different ( χ2=43.82, P<0.001); the male to female ratio was 1∶1.14 ( χ2=1.63, P=0.442); 3 317 cases (51.26%, 3 317/6 471) were injured by cats, and 2 614 cases (42.16%, 2 614/6 471) were injured by dogs, cats and dogs were the main injured animals ( χ2=18.63, P=0.098). The upper limbs (4 131/6 471, 63.84%) and lower limbs (1 848/6 471, 28.56%) were the most exposed sites, and there was a statistically significant difference in the exposure composition ratio of each site in three years ( χ2=105.79, P<0.001). Grade II exposure accounted for 31.79% (2 057/6 471), grade III exposure accounted for 62.31% (4 032/6 471). Among grade III exposure individuals, those who used passive immune preparations accounted for 55.13% (2 232/4 032). Conclusions:The number of rabies-exposed patients in the animal injury outpatient department of Lishui Central Hospital has been increasing year by year, and the population of patients injured by cats and dogs is particularly prominent. The utilization rate of passive immune preparations for grade III exposure patients still needs to be further improved.

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.

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 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.

The successful suppression of clutter arising from stationary or slowly moving tissue is one of the key issues in medical ultrasound color blood imaging. Remaining clutter may cause bias in the mean blood frequency estimation and results in a potentially misleading description of blood-flow. In this paper, based on the principle of general wall-filter, the design process of three classes of filters, infinitely impulse response with projection initialization (Prj-IIR), polynomials regression (Pol-Reg), and eigen-based filters are previewed and analyzed. The performance of the filters was assessed by calculating the bias and variance of a mean blood velocity using a standard autocorrelation estimator. Simulation results show that the performance of Pol-Reg filter is similar to Prj-IIR filters. Both of them can offer accurate estimation of mean blood flow speed under steady clutter conditions, and the clutter rejection ability can be enhanced by increasing the ensemble size of Doppler vector. Eigen-based filters can effectively remove the non-stationary clutter component, and further improve the estimation accuracy for low speed blood flow signals. There is also no significant increase in computation complexity for eigen-based filters when the ensemble size is less than 10.
Blood Flow Velocity ; Color ; Humans ; Signal Processing, Computer-Assisted ; Ultrasonography