A novel dual-target Septin9 methylation assay for improved detection of early-stage colorectal cancer and high-grade intraepithelial neoplasia.

BACKGROUND: Colorectal cancer (CRC) ranks as the third most common malignancies in the world, and periodic examination of the patient is advantageous in reducing the mortality of CRC. The first blood-based Septin9 gene methylation assay which recognized by the US FDA for CRC examination was Epi proColon. However, this assay was not broadly applied in the current clinical guideline because of its relatively lower sensitivity in the detection of early-stage CRC. METHODS: This study aimed at developing a new multiplex Septin9 methylation assay (ColonUSK) which simultaneously evaluates two CpG-rich subregions in the promoter of the Septin9 gene and an internal control in a single reaction. ColonUSK proved increased sensitivity, with a detection limit as low as 12pg of the positive DNA compared with the Septin9 assay targeting one CpG-rich subregion. 1366 subjects were prospectively recruited from four comprehensive hospitals in China in an opportunistic screening study for assessing its value in CRC detection. Blind testing was developed to evaluate ColonUSK in comparison with clinical examination using clinical gold standard such as colonoscopy. RESULTS: The assay demonstrates clinical sensitivity for diagnosing colorectal cancer (CRC) and advanced adenoma at rates of 77.34% and 25.26%, respectively. Furthermore, ColonUSK exhibits a high degree of specificity for non-CRC cases (95.95%) clinically. Significantly, the detection rate of cases in high-grade intraepithelial neoplasia increased to 54.29%. The value for the assay in the Kappa test was 0.76, showing a high degree of consistency between ColonUSK and clinical gold standard. CONCLUSIONS: ColonUSK indicated moderate diagnostic value and could become a non-invasive detection way for CRC. The implementation of the ColonUSK assay has the capacity to markedly enhance CRC screening practices.

Causes and Counter Measure Analysis of Inpatients Unqualified Clinical Specimens from 2015 to 2019.

BACKGROUND: To improve the quality of pre-analytical phase and provide targeted suggestions, this study analyzed factors causing unqualified clinical specimens in patients of the Department of Clinical Laboratory of Renmin Hospital of WuHan University from 2015 to 2019. METHODS: Inpatient specimens from January 2015 to December 2019 were retrospectively analyzed. Unqualified specimens were identified by referring to the general principle of rejection. The analytical indicators included incidence rate of unqualified specimens and constituent ratio of reasons of unqualified specimens. These two indicators were analyzed according to the inpatient wards and types of specimens. RESULTS: From 2015 to 2019, 21,674 inpatient unqualified specimens were collected, the incidence rate of unqualified specimens was 0.22% (21,674/9,700,869), the number and rate of unqualified specimens decreased year by year. The main reasons of unqualified specimens were insufficient volume (29.67%, 6,430/21,674) and clotting (26.31%, 5,703/21,674). The number of unqualified specimens in the departments of cardiovascular, pediatrics, neurology, oncology, urinary surgery, and intensive care unit ranked the top each year. Clotting (39.29%, 5,682/14,462) was the main reason of unqualified blood specimens while insufficient volume (71.18%, 3,365/4,727) was for urine specimens. Wrong identification caused unqualified feces (62.65%, 728/1,162) and body fluid (40.74%, 539/1,323) specimens. CONCLUSIONS: Clinical laboratory could make effective measures to improve pre-analytical quality by retrospectively analyzing data of unqualified specimens.

The Association Among Serum Growth Differentiation Factor 15 Level and Suicidal Ideation is Dependent on Testosterone Deficiency in Depressive Patients.

BACKGROUND: Elevated levels of growth differentiation factor (GDF)-15 and reduced levels of testosterone have been linked to depressive disorder, but whether they are also linked to suicidal ideation in patients with depression is unclear. METHODS: This retrospective study involved 301 patients ≥22 years old hospitalized for depression between July 2018 and November 2020 at Renmin Hospital of Wuhan University, of whom 120 had suicidal ideation. Serum levels of GDF-15 and testosterone were compared between patients with or without suicidal ideation. RESULTS: GDF-15 levels were significantly higher among patients with suicidal ideation than among those without, regardless of whether testosterone levels were above or below the median of 314 ng/dL. In multivariate logistic regression involving all patients, serum GDF-15 level was associated with low testosterone level (P=0.001). Among patients with testosterone <314 ng/dL, an increase of 1 standard deviation in serum GDF-15 level translated to a 1.3-fold increase in the risk of suicidal ideation (P=0.007). This relationship was not observed in all population or patients with testosterone levels ≥314 ng/dL. CONCLUSION: High serum GDF-15 level may be associated with an increased risk of suicidal ideation in patients with depression, and this association appears to be partly mediated by low testosterone levels.

A retrospective analysis of 12,400 SARS-CoV-2 RNA tests in patients with COVID-19 in Wuhan.

ABSTRACT: The outbreak and widely spread of coronavirus disease 2019 (COVID-19) has become a global public health concern. COVID-19 has caused an unprecedented and profound impact on the whole world, and the prevention and control of COVID-19 is a global public health challenge remains to be solved. The retrospective analysis of the large scale tests of SARS-CoV-2 RNA may indicate some important information of this pandemic. We selected 12400 SARS-CoV-2 tests detected in Wuhan in the first semester of 2020 and made a systematic analysis of them, in order to find some beneficial clue for the consistent prevention and control of COVID-19.SARS-CoV-2 RNA was detected in suspected COVID-19 patients with real-time fluorescence quantitative PCR (RT-qPCR). The patients' features including gender, age, type of specimen, source of patients, and the dynamic changes of the clinical symptoms were recorded and statistically analyzed. Quantitative and qualitive statistical analysis were carried out after laboratory detection.The positive rate of SARS-CoV-2 was 33.02% in 12,400 suspected patients' specimens in Wuhan at the first months of COVID-19 epidemics. SARS-CoV-2 RT-qPCR test of nasopharyngeal swabs might produce 4.79% (594/12400) presumptive results. The positive rate of SARS-CoV-2 RNA was significantly different between gender, age, type of specimen, source of patients, respectively (P < .05). The median window period from the occurrence of clinical symptom or close contact with COVID-19 patient to the first detection of positive PCR was 2 days (interquartile range, 1-4 days). The median interval time from the first SARS-CoV-2 positive to the turning negative was 14 days (interquartile range, 8-19.25 days).This study reveals the comprehensive characteristics of the SARS-CoV-2 RNA detection from multiple perspectives, and it provides important clues and may also supply useful suggestions for future work of the prevention and treatment of COVID-19.

Diagnostic and Prognostic Potentials of Long Noncoding RNA ELF3-AS1 in Glioma Patients.

OBJECTIVE: Accumulating evidence implies that long noncoding RNAs (lncRNAs) play a crucial role in predicting survival for glioma patients. However, the potential function of lncRNA ELF3-antisense RNA 1 (ELF3-AS1) in tumors remained largely unclear. The aim of this study was to explore the expression of lncRNA ELF3-antisense RNA 1 (ELF3-AS1) and evaluate its functions in glioma patients. Patients and Methods. ELF3-AS1 expressions were examined by RT-PCR in 182 pairs of glioma specimens and adjacent normal tissues. The receiver operating characteristic (ROC) curve was performed to estimate the diagnostic value of ELF3-AS1. The chi-square tests were used to examine the associations between ELF3-AS1 expression and the clinicopathological characters. The overall survival (OS) and disease-free survival (DFS) were analyzed by log-rank test, and survival curves were plotted according to Kaplan-Meier. The prognostic value of the ELF3-AS1 expression in glioma patients was further analyzed using univariate and multivariate Cox regression analyses. Loss-of-function assays were performed to determine the potential function of ELF3-AS1 on the proliferation and invasion of glioma cells. RESULTS: The ELF3-AS1 expression level was significantly higher in glioma specimens compared with adjacent nontumor specimens (p < 0.01). A high expression of ELF3-AS1 was shown to be associated with the WHO grade (p = 0.023) and KPS score (p = 0.012). ROC assays revealed that high ELF3-AS1 expression had an AUC value of 0.8073 (95% CI: 0.7610 to 0.8535) for glioma. Using the Kaplan-Meier analysis, we found that patients with a high ELF3-AS1 expression had significantly poor OS (p = 0.006) and DFS (p = 0.0002). In a multivariate Cox model, we confirmed that ELF3-AS1 expression was an independent poor prognostic factor for glioma patients. The functional assay revealed that knockdown of ELF3-AS1 suppressed the proliferation and invasion of glioma cells. CONCLUSIONS: Our findings confirmed that ELF3-AS1 functions as an oncogene in glioma and indicated that ELF3-AS1 is not only an important prognostic marker but also a potential therapy target for glioma.