DNA Methylation-Based Testing in Peripheral Blood Mononuclear Cells Enables Accurate and Early Detection of Colorectal Cancer.

An effective blood-based method for the diagnosis of colorectal cancer has not yet been developed. Molecular alterations of immune cells occur early in tumorigenesis, providing the theoretical underpinning for early cancer diagnosis based on immune cell profiling. Therefore, we aimed to develop an effective detection method based on peripheral blood mononuclear cells (PBMC) to improve the diagnosis of colorectal cancer. Analysis of the genome-wide methylation landscape of PBMCs from patients with colorectal cancer and healthy controls by microarray, pyrosequencing, and targeted bisulfite sequencing revealed five DNA methylation markers for colorectal cancer diagnosis, especially early-stage colorectal cancer. A single-tube multiple methylation-specific quantitative PCR assay (multi-msqPCR) for simultaneous detection of five methylation markers was established, which allowed quantitative analysis of samples with as little as 0.1% PBMC DNA and had better discriminative performance than single-molecule detection. Then, a colorectal cancer diagnostic model (CDM) based on methylation markers and the multi-msqPCR method was constructed that achieved high accuracy for early-stage colorectal cancer (AUC = 0.91; sensitivity = 81.18%; specificity = 89.39%), which was improved compared with CEA (AUC = 0.79). The CDM also enabled a high degree of discrimination for advanced adenoma cases (AUC = 0.85; sensitivity = 63.04%). Follow-up data also demonstrated that the CDM could identify colorectal cancer potential up to 2 years before currently used diagnostic methods. In conclusion, the approach constructed in this study based on PBMC-derived DNA methylation markers and a multi-msqPCR method is a promising and easily implementable diagnostic method for early-stage colorectal cancer. SIGNIFICANCE: Development of a diagnostic model for early colorectal cancer based on epigenetic analysis of PBMCs supports the utility of altered DNA methylation in immune cells for cancer diagnosis.

A multiplex blood-based assay targeting DNA methylation in PBMCs enables early detection of breast cancer.

The immune system can monitor tumor development, and DNA methylation is involved in the body's immune response to tumors. In this work, we investigate whether DNA methylation alterations in peripheral blood mononuclear cells (PBMCs) could be used as markers for early detection of breast cancer (BC) from the perspective of tumor immune alterations. We identify four BC-specific methylation markers by combining Infinium 850 K BeadChips, pyrosequencing and targeted bisulfite sequencing. Based on the four methylation markers in PBMCs of BC, we develop an efficient and convenient multiplex methylation-specific quantitative PCR assay for the detection of BC and validate its diagnostic performance in a multicenter cohort. This assay was able to distinguish early-stage BC patients from normal controls, with an AUC of 0.940, sensitivity of 93.2%, and specificity of 90.4%. More importantly, this assay outperformed existing clinical diagnostic methods, especially in the detection of early-stage and minimal tumors.

Acupuncture treatment of glaucoma based on radar plots: A protocol for an overview of systematic reviews.

BACKGROUND: Glaucoma is the second most blinding eye disease in the world. Currently, lowering the intraocular pressure through various methods is the main treatment of glaucoma. Acupuncture has been effectively and safely used in the treatment of glaucoma. However, the evidence for the efficacy of acupuncture in the treatment of glaucoma is controversial, leading to inconsistent findings from systematic evaluations at abroad and home. Therefore, this protocol aims to provide a multivariate evaluation on the quality of evidences from current systematic reviews (SRs) and/or meta-analyzes (MAs) of acupuncture in the treatment of glaucoma, and literature quality, thus providing an intuitive and reliable evidence synthesis and basis for clinical decision making. METHODS: MAs/SRs about the acupuncture treatment of glaucoma will be searched online, including Chinese Biomedical Literature Database (CBM), China Science and Technology Journal Database (VIP), China National Knowledge Infrastructure (CNKI), WanFang Database (WF), Web of Science, Embase, PubMed, and Cochrane Library. Two investigators will independently screen literatures according to inclusion and exclusion criteria and extract data. A multivariate evaluation of the included literature will be performed by depicting radar plots in 6 aspects as follows: Year of publication, study type, SRs assessment through the Assessment of Multiple Systematic Reviews 2 (AMSTAR 2), literature quality assessment through the Preferred Reporting Items for Systematic Reviews and Meta-analyses (PRISMA), homogeneity, and publication bias. The Grading of Recommendations Assessment, Development, and Evaluation evidence quality assessment tool will be used to grade and evaluate the quality of outcome indicators of the included literatures. RESULTS: This study will be submitted for publication in a peer-reviewed journal. CONCLUSION: We would like to provide a visual and scientific approach for clinical decision making of acupuncture treatment of glaucoma through a accessible and useful assessment of systematic reviews.

Astragalus â £ ameliorates the dry eye injury in rabbit model via MUC1-ErbB1 pathway.

The therapeutic effects and potential mechanisms of astragaloside IV on a rabbits dry eye model induced by benzalkonium chloride (BAC) was examined. In our study, a BAC-induced dry eye rabbit model was treated with eye drops containing astragaloside IV (5, 10 µM) or solvent four times a day. The clinical evaluations, such as tear break-up time (BUT) and Schirmer tear test (STT), were performed on days 0, 7, 14, 21, and 28. On day 28, the cornea and bulbar conjunctiva tissues (left eye and right eye) were collected with histology, and immunofluorescent staining conducted. The levels of MUC1 and ErbB1in the corneas were determined by real-time quantitative PCR (qRT-PCR) and the proteins levels of MUC1 and ErbB1 were detected by Western blot. It was demonstrated that both astragaloside IV (5, 10 µM) treatments resulted in an increased STT and BUT on days 7, 14, 21 and 28. Additionally, the astragaloside IV (5, 10 µM)-treated group showed increasing PAS-positive goblet cells than model group (0 µM). Moreover, the MUC1 in model group (0 µM) was decreased, while the expression of MUC1 in astragaloside IV (5, 10 µM) group was increased. Furthermore, astragaloside IV had a protective effect on BAC-induced rabbits' dry eye and demonstrated clinical improvements, which indicated that astragaloside IV served as a potential protective agent in the clinical treatment of dry eye.