Evaluation of the analytical performance of endocrine analytes using sigma metrics.

BACKGROUND: (a) To evaluate the clinical performance of endocrine analytes using the sigma metrics (σ) model. (b) To redesign quality control strategies for performance improvement. METHODS: The sigma values of the analytes were initially evaluated based on the allowable total error (TEa), bias, and coefficient of variation (CV) at QC materials level 1 and 2 in March 2018. And then, the normalized QC performance decision charts, personalized QC rules, quality goal index (QGI) analysis, and root causes analysis (RCA) were performed based on the sigma values of the analytes. Finally, the sigma values were re-evaluated in September 2018 after a series of targeted corrective actions. RESULTS: Based on the initial sigma values, two analytes (FT3 and TSH) with σ > 6, only needed one QC rule (13S ) with N2 and R500 for QC management. On the other hand, seven analytes (FT4, TT4, CROT, E2, PRL, TESTO, and INS) with σ < 4 at one QC material level or both needed multiple rules (13S /22S /R4S /41S /10X ) with N6 and R10-500 depending on different sigma values for QC management. Subsequently, detailed and comprehensive RCA and timely corrective actions were performed on all the analytes base on the QGI analysis. Compared with the initial sigma values, the re-evaluated sigma metrics of all the analytes increased significantly. CONCLUSIONS: It was demonstrated that the combination of sigma metrics, QGI analysis, and RCA provided a useful evaluation system for the analytical performance of endocrine analytes.

Technical and Clinical Performance of Two Methods to Detect Squamous Cell Carcinoma Antigen Levels for Comparing Pathological Diagnosis Coincidence Rates in Lung, Cervical, and Head and Neck Cancers.

BACKGROUND: Detection of serum squamous cell carcinoma (SCC) antigen (SCCA) contributes to the diagnosis and therapeutic monitoring of SCC. However, the results obtained from different detection systems are not always consistent. METHODS: In this study, we compared the performance of electrochemiluminescence assays (ECLIAs) and flow fluorescence immunoassays (FFIAs) (e.g. using a Luminex 200/xMap) in the detection of SCCA for the diagnosis of SCC of the lung (LSCC), cervix (CSCC), and head and neck (HANSCC) in serum samples from 154 healthy individuals and 236 patients with SCC. We also evaluated the consistency of the SCCA results with the pathological diagnosis for both methods. RESULTS: SCCA levels obtained from ECLIAs were significantly higher than those obtained from FFIAs for all groups. However, the results from the two methods were well correlated (r = 0.918). The diagnostic coincidence rates (FFIA versus ECLIA) for SCCA results in patients with LSCC, CSCC, and HANSCC were 40.82% versus 52.04%, 36.14% versus 57.14%, and 16.36% versus 23.64%, respectively, and the negative coincidence rate (FFIA versus ECLIA) in healthy individuals was 98.05% versus 98.70%. The cutoff value, sensitivity, specificity, and area under the receiver operating characteristic curve of SCCA diagnosis (FFIA versus ECLIA) in LSCC, CSCC, and HANSCC were 1.12, 77.55%, 85.34%, and 0.87 versus 3.07, 85.71%, 91.52%, and 0.91, respectively; 1.21, 81.93%, 90.2%, and 0.90 versus 3.84, 89.16%, 95.24%, and 0.95, respectively; and 1.01, 62.27%, 82%, and 0.81 versus 3.35, 58.18%, 89.65%, and 0.85, respectively. CONCLUSIONS: Serum SCCA levels detected by ECLIAs were significantly higher than those detected by FFIAs, with higher detection performance and pathological diagnosis coincidence rate in the patients with LSCC and CSCC simultaneously.

Aberrant expression of two miRNAs promotes proliferation, hepatitis B virus amplification, migration and invasion of hepatocellular carcinoma cells: evidence from bioinformatic analysis and experimental validation.

BACKGROUND: As key negative regulators of gene expression, microRNAs (miRNAs) play an important role in the onset and progression of hepatocellular carcinoma (HCC). This study aimed to identify the miRNAs involved in HCC carcinogenesis and their regulated genes. METHODS: The Gene Expression Omnibus (GEO) dataset (GSE108724) was chosen and explored to identify differentially expressed miRNAs using GEO2R. For the prediction of potential miRNA target genes, the miRTarBase was explored. Enrichment analysis of Gene Ontology (GO) and the Kyoto Encyclopedia of Genes and Genomes (KEGG) was performed by the DAVID online tool. The hub genes were screened out using the CytoHubba plug-in ranked by degrees. The networks between miRNAs and hub genes were constructed by Cytoscape software. MiRNA mimics and negative control were transfected into HCC cell lines and their effects on proliferation, hepatitis B virus DNA (HBV-DNA) replication, TP53 expression, migration, and invasion were investigated. The following methods were employed: MTT assay, quantitative PCR (qPCR) assay, western blotting, wound healing assay, and transwell assay. RESULTS: A total of 50 differentially expressed miRNAs were identified, including 20 upregulated and 30 downregulated miRNAs, in HCC tumor tissues compared to matched adjacent tumor-free tissues. The top three upregulated (miR-221-3p, miR-222-3p, and miR-18-5p) and downregulated (miR-375, miR-214-3p and miR-378d) miRNAs, ranked by |log2 fold change (log2FC)|, were chosen and their potential target genes were predicted. Two gene sets, targeted by the upregulated and the downregulated miRNAs, were identified respectively. GO and KEGG pathway analysis showed that the predicted target genes of upregulated and downregulated miRNAs were mainly enriched in the cell cycle and cancer-related pathways. The top ten hub nodes of gene sets ranked by degrees were identified as hub genes. Analysis of miRNA-hub gene network showed that miR-221-3p and miR-375 modulated most of the hub genes, especially involving regulation of TP53. The q-PCR results showed that miR-221-3p and miR-375 were markedly upregulated and downregulated, respectively, in HCC cells and HCC clinical tissue samples compared to non-tumoral tissues. Furthermore, miR-221-3p overexpression significantly enhanced proliferation, HBV-DNA replication, as well as the migration and invasion of HCC cells, whereas miR-375 overexpression resulted in opposite effects. Western blotting analysis showed that the overexpression of miR-221-3p and miR-375 reduced and increased TP53 expression, respectively. CONCLUSION: The present study revealed that miR-211-3p and miR-375 may exert vital effects on cell proliferation, HBV-DNA replication, cell migration, and invasion through the regulation of TP53 expression in HCC.

[Determination of oxidation-reduction level in patients with rheumatoid arthritis].

OBJECTIVE: To examine the oxidation or reduction products in patients with rheumatism arthritis (RA), and investigate the relationship between oxidation or reduction products and occurrence and development of RA. METHODS: The serum levels of total ascorbic acid (TAA), dehydroascorbic acid (DHAA)/TAA, vitamin E, advanced oxidation protein products (AOPP) and malondialdehyde (MDA) were detected by high-performance liquid chromatography with electrochemical detection in 83 RA patients and 30 healthy adults. Correlation analysis of AOPP, MDA and hs-CRP was performed. RESULTS: Compared with normal control group, significantly higher serum MDA, DHAA/TAA, and AOPP levels were detected in RA patients (P<0.05), but vitamin E showed no significant difference (P<0.05). Linear regression analysis showed that MDA (P<0.01) was positively but AOPP (P>0.05) negatively correlated to hs-CRP. CONCLUSIONS: Oxidation or reduction products in serum of RA patients increases significantly, which may be an important mechanism for the occurrence and development of RA. Serum AOPP and MDA levels can reflect the oxidation status in RA patients.