Plasma cell-free DNA methylome-based liquid biopsy for accurate gastric cancer detection.

Early detection plays a critical role in mitigating mortality rates linked to gastric cancer. However, current clinical screening methods exhibit suboptimal efficacy. Methylation alterations identified from cell-free DNA (cfDNA) present a promising biomarker for early cancer detection. Our study focused on identifying gastric cancer-specific markers from cfDNA methylation to facilitate early detection. We enrolled 150 gastric cancer patients and 100 healthy controls in this study, and undertook genome-wide methylation profiling of cfDNA using cell-free methylated DNA immunoprecipitation and high-throughput sequencing. We identified 21 differentially methylated regions (DMRs) between the gastric tumor and nontumor groups using multiple algorithms. Subsequently, using the 21 DMRs, we developed a gastric cancer detection model by random forest algorithm in the discovery set, and validated the model in an independent set. The model was able to accurately discriminate gastric cancer with a sensitivity and specificity of 93.90% and 95.15% in the discovery set, respectively, and 88.38% and 94.23% in the validation set, respectively. These results underscore the efficacy and accuracy of cfDNA-derived methylation markers in distinguishing early stage gastric cancer. This study highlighted the significance of cfDNA methylation alterations in early gastric cancer detection.

Effect of Laparoscopy Combined with Choledochoscope for the Treatment of Cholecystolithiasis and Choledocholithiasis.

Objective: Evaluate the influence of laparoscopy combined with choledochoscopy on operation-related indexes, serum total bilirubin (TBIL) level, and abdominal drainage tube extraction time within cases carrying cholecystolithiasis/choledocholithiasis. Methods: 86 cases of cholecystolithiasis together with choledocholithiasis were chosen for this investigation, and cases were randomly segregated within the control cohort (43 cases, open surgery) and observation cohort (43 cases, laparoscopy combined with choledochoscopy).The operation-related indexes, complete stone clearance rate, postoperative visual analogue scale (VAS) scoring, serum TBIL level, and postsurgical complications/recovery incidence were observed and comparatively analyzed across cohorts. Results: Compared with the control cohort, the incision length, operation duration, postoperative exhaust duration, abdominal drainage tube extraction time, and postsurgery hospitalization in observation cohort were markedly reduced (P < 0.05), the intrasurgical hemorrhaging was markedly reduced (P < 0.05), and the postoperative complication incidences were markedly reduced (P < 0.05). Furthermore, the complete stone clearance rates in the observation cohort were elevated compared with control, but the difference was not statistically significant (P > 0.05).VAS scoring for the observation cohort at 6, 12, 24, and 48 hours postsurgery was markedly reduced (P < 0.05). On the first day after the operation, the serum TBIL levels for the two cohorts were very high and gradually decreased, and the serum TBIL levels in the observation cohort were markedly reduced during day 1, 3, and 5 postsurgery (P < 0.05). Conclusion: Laparoscopy combined with choledochoscopy surgical treatment might reduce the surgery duration, intrasurgery hemorrhaging, postsurgical pain, and liver function damage.

Enrichment and detection of circulating tumor cells by immunomagnetic beads and flow cytometry.

OBJECTIVE: The purpose of the article is to establish a quick enrichment and detection method using immunomagnetic beads and flow cytometry to analyze circulating tumor cells (CTCs) in the peripheral blood. RESULTS: After incubation with CD326-PE and CD45-APC antibodies, more than 60% MCF7 cells in M-Buffer could be detected while less than 10% of the same cells could be detected by flow cytometry (FCM) if spiked into blood. However, in combination with CD326 and CD45 immunomagnetic beads, detection rate of MCF7 cells in blood reached 57%. For circulating tumor cells, enrichment by CD326 and CD45 immunomagnetic beads improve the detection rate from nearly undetectable to more than 24.14%. CONCLUSIONS: Live CTCs in peripheral blood can be effectively and sensitively detected by using a combination of immunomagnetic beads (CD45 and CD326) and flow cytometry.

Epithelial-mesenchymal transition classification of circulating tumor cells in lung and colon cancer patients: potential role in clinical practice.

BACKGROUND: Tumor cells undergoing epithelial-mesenchymal transition (EMT) display enhanced ability to enter the circulation, thereby being major source of circulating tumor cells (CTCs). In this study, we aimed to better understand the roles of CTC undergoing EMT in monitoring cancer progression. METHODS: We analyzed gene expression profiling of epithelial and mesenchymal markers in lung or colon tumor samples by mining TCGA database. We detected CTCs and classify their EMT phenotypes of 31 patients with lung or colon cancer by using a CanPatrol CTC-enrichment technique. RESULTS: The bioinformatic analysis indicated that mesenchymal markers were expressed in a subset of lung tumor samples, and its high expression was associated with poor survival of lung cancer patients. However, in colon cancer, majority of tumor samples expressed hybrid epithelial/mesenchymal markers. CTC analysis with EMT classification showed that the number of CTCs with mesenchymal phenotype was high in lung cancer patients with the advanced stage. Dynamic CTC analysis in a lung cancer patient indicated that CTC with mesenchymal phenotype was effective to monitor tumor progression. In a colon cancer patient, dynamic CTC analysis indicated that CTC with hybrid epithelial/mesenchymal phenotypes was an effective biomarker to guide therapy. CONCLUSIONS: Encouraging results from this proof-of-concept study show that CTC with mesenchymal phenotype or hybrid epithelial/mesenchymal phenotypes could be a potential biomarker for monitoring tumor progression in lung or colon cancer respectively.