A sensitive and robust plasma-based DNA methylation panel for early detection of target gastrointestinal cancers.

Background: Target gastrointestinal cancers (GICs), encompassing esophageal cancer (EC), gastric cancer (GC), and colorectal cancer (CRC), originate within a single readily accessible luminal organ system and are diagnosable using endoscopy. However, endoscopy is an invasive procedure with low compliance and no plasma-based DNA methylation assay for the early detection of GICs.

Methods: Nine potential DNA methylation markers were identified and evaluated in tissue (n=60) and plasma (n=155) cohorts to select the most suitable markers.

Identifying potential DNA methylation markers for the detection of esophageal cancer in plasma.

Esophageal cancer (EC) is a leading cause of cancer-related deaths in China, with the 5-year survival rate reaching less than 30%, because most cases were diagnosed and treated at the advanced stage. However, there is still a lack of low-cost, efficient, and accurate non-invasive methods for the early detection of EC at present. A total of 48 EC plasma and 101 control plasma samples were collected in a training cohort from 1 January 2021 to 31 December 2021, and seven cancer-related DNA methylation markers (, , , , , and ) were tested in these samples to select potential markers.

Improved PCR by the Use of Disruptors, a New Class of Oligonucleotide Reagents.

As a powerful tool, polymerase chain reaction (PCR) has been indispensable and widely used in a large array of applications. In practice, many factors may affect the overall performance of a PCR. One such factor is the stability of intramolecular secondary structure formed within single-stranded template.

ANXA2 as a novel substrate of FBXW7 promoting esophageal squamous cell carcinoma via ERK phosphorylation.

Our recent study suggests that FBXW7 loss of function plays a critical function in esophageal cancer. However, the mechanism of FBXW7 in promoting esophageal cancer is still unclear. Here, we explored the interaction protein of FBXW7 by screening of GST-pulldown and LC-MS/MS analysis in esophageal squamous cell carcinoma (ESCC) and identified ANXA2 as a potential target of FBXW7.

Real-world application of a fast stool DNA test for colorectal cancer screening in primary screening positive population.

Background: Stool DNA test has been emerged as an effective noninvasive method for colorectal cancer (CRC) screening, but the real-world performance of stool DNA test in Chinese population has rarely been reported.

Methods: A total of 36,527 subjects were recruited in Haining City from January 2021 to December 2021. Participants underwent primary screening by taking both two-samples fecal immunochemical tests (FITs) and high-risk factor questionnaire (HRFQ), and those who tested either positive by FITs or evaluated to be high risk by HRFQ were recommended to undertake subsequent stool DNA test and colonoscopy.

A Novel Plasma-Based Methylation Panel for Upper Gastrointestinal Cancer Early Detection.

Background: Upper gastrointestinal cancer (UGC) is an important cause of cancer death in China, with low five-year survival rates due to the majority of UGC patients being diagnosed at an advanced stage. Therefore, there is an urgent need to develop cost-effective, reliable and non-invasive methods for the early detection of UGC.

Methods: A novel plasma-based methylation panel combining simultaneous detection of three methylated biomarkers (, and ) and an internal control gene were developed and used to examine plasma samples from 186 UGC patients and 190 control subjects.

A simplified multiplex methylated DNA testing for early detection of colorectal cancer in stool DNA.

Background: ColoDefense1.0 assay has demonstrated its excellent sensitivity and specificity for early detection of colorectal cancer (CRC) by detecting the methylation levels of SDC2 and SEPT9, while exhibited limitations on relatively large sample capacity required and limited detection throughput by applying triplicate PCR reactions for each sample. In this study, ColoDefense1.

Feasibility and reproducibility of a plasma-based multiplex DNA methylation assay for early detection of gastric cancer.

Background: Gastric cancer (GC) is a leading cause of cancer death and an important barrier to increasing life expectancy in China. Early detection of GC can significantly reduce its mortality rate.

Methods: A new plasma-based multiplex DNA methylation assay combining simultaneous detection of three biomarkers (KCNQ5, C9orf50 and CLIP4) and one control gene (ACTB) was developed.

Rapid cultivation of anammox sludge based on Ca-alginate cell beads.

Current gel entrapment technology has certain advantages for the enrichment of anammox sludge. In this study, the optimal preparation conditions and cultivation equipment of Ca-alginate cell beads for the culturing anammox sludge were proposed. The preparation parameters of the Ca-alginate cell beads were as follows: 3% sodium alginate, 4% CaCl, V:V = 1:1, a drop height of 9 cm, stirring speed of 300 rpm, and cross-linking time of 24 h.

Surface modified silver/magnetite nanocomposite activating hydrogen peroxide for efficient degradation of chlorophenols.

The mixed-valent magnetite (FeO) played a critical role in HO-based Fenton-like system for the removal of chlorophenols, but high activity and cycle stability of the FeO-based catalysts are still a huge challenge. Herein, a series of surface hydroxyl- and carboxyl-modified Ag/FeO nanocomposite catalysts were prepared and used to activate HO for degradation chlorophenols pollutants. Under the optimized condition, nearly 100% degradation ratio were achieved within 2-30 min for 2,4-dichlorophenol, 2,3-dichlorophenol, 3,4-dichlorophenol, 2,4,6-trichlorophenol, p-nitrophenol, and 98% degradation ratio for 2,5-dichlorophenol, 2,6-dichlorophenol and 3,5-dichlorophenol,.

Blood leukocytes methylation levels analysis indicate methylated plasma test is a promising tool for colorectal cancer early detection.

: A number of plasma methylated DNA biomarkers related to colorectal cancer (CRC) have been identified. However, the effect of methylation level in leukocytes on plasma-based methylation test was rarely reported. : Blood samples from 213 individuals including 91 CRC patients were collected and separated into 3.

Combining Serum DNA Methylation Biomarkers and Protein Tumor Markers Improved Clinical Sensitivity for Early Detection of Colorectal Cancer.

Background: Colorectal cancer (CRC) is one of the leading causes of cancer deaths worldwide and in China. Early CRC screening is the best approach to reduce its incidence and mortality rates. The ColoDefense test, a multiplex qPCR assay simultaneously detecting both methylated and genes, has demonstrated improved clinical performance on either methylation biomarker alone for CRC screening with both blood and stool samples.

Feasibility of Methylated in Stool for Early Detection of Colorectal Cancer: A Training Study in Chinese Population.

Background: Early detection of colorectal cancer (CRC) and precancerous lesion is vitally important for mitigating CRC morbidity and mortality. Aberrant DNA methylations in certain promoter regions have been identified to be closely associated with CRC development and progression, suggesting their potential as diagnostic biomarkers for early detection. In this study, we evaluated the performance of methylated in stool specimens as a potential biomarker for CRC detection.

Methylated and in stool specimens for Colorectal Cancer early detection: A cost-effective strategy for Chinese population.

The aim of this study was to evaluate the feasibility of combination of methylated and methylated (SpecColon test) in stool specimens for colorectal cancer (CRC) early detection and to optimize the cut-off value of methylated and methylated . Approximately 5 g of stool specimen each was collected from 420 subjects (291 in the training cohort and 129 in the validation cohort). Stool DNA was extracted and bisulfite-converted, followed by detection of methylated level of and .

Disruptors, a new class of oligonucleotide reagents, significantly improved PCR performance on templates containing stable intramolecular secondary structures.

Intramolecular secondary structures within templates have been shown to lower PCR performance. Whereas many approaches have been developed to mitigate such impairment on PCR, their effects can vary greatly depending on template sequences. Here we present a novel, universally effective approach to improve PCR performance involving specifically designed oligonucleotides called disruptors.

Transient stem-loop structure of nucleic acid template may interfere with polymerase chain reaction through endonuclease activity of Taq DNA polymerase.

As a standard molecular biology technique, PCR uses DNA polymerase to detect, amplify and manipulate DNA targets. Due to its effect of exponential amplification, PCR can achieve high sensitivity required for detecting targets of low abundance. Therefore, it has become the method of choice for the majority of nucleic acid-based tests.

Aberrant DNA Methylation of and in Stool Specimens as an Integrated Biomarker for Colorectal Cancer Early Detection.

Colorectal cancer (CRC) has become the second leading cause of new cancer cases and the fifth of cancer deaths in China, and early detection is the most effective way to reduce the incidence and mortality of CRC. A number of methylated DNA biomarkers have been found to associate with CRC and precancerous lesions in stool samples, indicating stool methylated DNA biomarkers are potential tools for CRC early detection. In this study, approximately 5 g of stool specimen was collected from 230 subjects (124 in the training set and 106 in the validation set).

Feasibility of Plasma-Methylated for Early Detection of Gastric Cancer.

Gastric cancer (GC) is fifth most frequently diagnosed cancer and second leading cause of cancer in China. More than 80% of GC are diagnosed at an advanced stage due to low uptake rate of invasive screening method. The performance of methylated test was evaluated in 236 plasma samples, including 92 patients with GC, 16 intestinal metaplasia patients, 26 gastric fundic gland polyp patients, 13 small adenoma patients, 39 hyperplastic polyp patients, and 50 control patients.

Performance Comparison Between Plasma and Stool Methylated Tests for Detecting Colorectal Cancer.

Colorectal cancer (CRC) is the most common type of malignancies of the gastrointestinal tract worldwide. Plasma methylated test has been used clinically for CRC screening for several years, but the study about the performance comparison between plasma and stool has rarely been reported. In this study, 124 plasma samples, 100 stool samples, and 60 sets of plasma and paired stool samples were collected and tested by a methylated test in three PCR replicates.

A novel plasma based early colorectal cancer screening assay base on methylated SDC2 and SFRP2.

Background: Methylated SFRP2 was previously reported as a non-invasive biomarker for colorectal cancer (CRC) detection with a relatively low sensitivity for early stage CRC. The purpose of this study was to evaluate the performance of a new plasma based CRC screening assay, SpecColon test, which tested methylated SFRP2 and SDC2 simultaneously in a single qPCR reaction, in detecting CRC and advanced adenomas (AA).

Method: One milliliter plasma of 122 CRC patients, 12 AA patients, 93 patients with benign polyps, and 91 normal individuals were collected from the Affiliated Hospital of Xuzhou Medical University, and all samples were examined by SpecColon test.

A cautionary tale of cross-contamination among plasmids from commercial suppliers.

Many researchers have switched to purchasing their desired plasmids from commercial suppliers to save time and resources, as we did for 17 high-risk human papillomavirus plasmids. To our surprise, they were shown to be cross-contaminated with one another. Comparison between the production schedule and the pattern of contaminations proved that this contamination occurred during the production process, which was also shown for another two sets of commercial plasmids.

Multiplex methylated DNA testing in plasma with high sensitivity and specificity for colorectal cancer screening.

Methylated SEPT9 showed relatively low sensitivity in detecting early stage colorectal cancer (CRC) and advanced adenomas (AA) in plasma. Combination of multiple biomarkers was an effective strategy to improve sensitivity in early stage cancer diagnosis and screening. A new qPCR-based assay combining the detection of methylated SEPT9 and SDC2 (ColoDefense test) was used.

Performance of a Novel Blood-Based Early Colorectal Cancer Screening Assay in Remaining Serum after the Blood Biochemical Test.

Background: Combination of multiple biomarkers was an effective strategy to improve sensitivity in cancer diagnosis and screening. However, the performance of the combination of methylated and for detection of colorectal cancer (CRC) has yet to be reported.

Methods: A new qPCR-based assay combining the detection of methylated and was used.

Performance of a MethyLight assay for methylated SFRP2 DNA detection in colorectal cancer tissue and serum.

Background: Colorectal cancer is one of the five most common cancers in China, and its incidence is steadily increasing. An accurate and non-invasive screening method is needed to increase the population uptake of colorectal cancer screening. Secreted frizzled-related protein 2 ( SFRP2) has been found to be hypermethylated in most colorectal cancer patients, and it may fulfill the role of a non-invasive biomarker for colorectal cancer screening.