Using signal-to-cutoff ratios of HIV screening assay to predict HIV infection.

BACKGROUND: The sensitivity of HIV screening assays often leads to a high rate of false-positive results, requiring retests and confirmatory tests. This study aimed to analyze the capability of signal-to-cutoff (S/CO) ratios of HIV screening assay to predict HIV infection. METHODS: A retrospective study on the HIV screening-positive population was performed at Zhongshan Hospital, Xiamen University, the correlation between HIV screening assay S/CO ratios and HIV infection was assessed, and plotted Receiver Operating Characteristic (ROC) curves were generated to establish the optimal cutoff value for predicting HIV infection. RESULTS: Out of 396,679 patients, 836 were confirmed to be HIV-infected, with an HIV prevalence of 0.21%. The median S/CO ratios in HIV infection were significantly higher than that in non-HIV infection (296.9 vs. 2.41, P < 0.001). The rate of confirmed HIV infection was increased with higher S/CO ratios in the screening assay. The ROC curve based on the HIV screening assay S/CO ratio achieved a sensitivity of 93.78% and a specificity of 93.12% with an optimal cutoff value of 14.09. The area under the ROC curve was 0.9612. Further analysis of the ROC curve indicated that the S/CO ratio thresholds yielding positive predictive values of 99%, 99.5%, and 100% for HIV infection were 26.25, 285.7, and 354.5, respectively. CONCLUSION: Using HIV screening assay S/CO ratio to predict HIV infection can largely reduce necessitating retests and confirmatory tests. Incorporating the S/CO ratio into HIV testing algorithms can have significant implications for medical and public health practices.

Significantly elevated serum human epididymis protein-4 in chronic kidney disease patients without ovarian cancer: A large-scale retrospective study.

BACKGROUNDS: Human epididymis protein-4 (HE-4) is a commonly used biomarker for diagnosing ovarian cancer. Elevated HE-4 has also been observed in various benign conditions including chronic kidney disease (CKD); however, generalizability and statistical power of previous studies have been limited by small sample sizes. MATERIALS AND METHODS: We conducted a retrospective study that included 80 pathologically confirmed ovarian cancer patients, 641 CKD patients, and 2661 healthy controls. Serum HE-4 and several renal function parameters were collected and compared between the three groups. Correlation analysis was conducted to evaluate the relationship between HE-4 and renal function parameters. A receiver operating characteristic curve was established to evaluate its diagnostic performance. RESULTS: CKD patients had the highest levels of HE-4, with a median of 193.00 pmol/L, while the median in ovarian cancer patients was 90.82 pmol/L. HE-4 levels also increased with CKD progression, and Spearman's rank correlation showed that HE-4 had a strong correlation with renal function parameters in CKD patients. Furthermore, HE-4 exhibited a satisfactory diagnostic performance in both differentiating CKD patients and controls as well as stage 2 CKD patients and controls. CONCLUSION: HE-4 can be used as an alternative biomarker for diagnosing CKD as it is less affected by several preanalytical factors. Nevertheless, in clinical practice, elevated HE-4 requires taking both CKD and ovarian cancer into consideration.

Assessment of anti-nucleosome antibody (ANuA) isotypes for the diagnosis and prediction of systemic lupus erythematosus and lupus nephritis activity.

Our study aims to investigate the serum levels of anti-nucleosome antibody (ANuA) isotypes in patients with systemic lupus erythematosus (SLE) and clarify ANuA isotypes that may diagnose and predict SLE. We detected anti-nucleosome antibodies (ANuA) in the serum from 120 patients with SLE, 99 patients suffering from other autoimmune diseases (OAD), and 50 healthy controls by performing IgG-, IgA-, and IgM-specific ELISAs. The serum levels of total anti-nuclear antibodies (ANA IgG), ANuA IgG subclasses (IgG1, IgG2, IgG3, and IgG4), anti-dsDNA antibodies, and the avidities of ANA IgG were also analysed using ELISAs. The levels of three ANuA isotypes (IgG, IgA, and IgM) were significantly higher in patients with SLE than in patients with OAD and healthy controls (p < 0.05). Moreover, the concentrations of ANuA isotypes increased in the active SLE and lupus nephritis (LN) groups and in patients with SLE presenting high-avidity IgG ANA (p < 0.05). Furthermore, ANuA isotype levels decreased significantly with drug therapy, while anti-dsDNA IgG levels decreased with the same trend. Additionally, ANuA isotypes were positively related to the SLEDAI (SLE Disease Activity Index) score, RAI (relative avidity index) of high-avidity IgG ANAs, and serum anti-dsDNA IgG levels. Last, the sensitivity and specificity values for SLE were 83.33 and 96.67% for ANuA IgG, 85.83 and 93.33% for ANuA IgA, and83.33 and 86.67% for ANuA IgM, respectively. The sensitivity and specificity values for LN were 61.67 and 96.67% for ANuA IgG, 49.17 and 96.67% for ANuA IgA, and 52.50 and 96.67% for ANuA IgM, respectively. In conclusion, we evaluated whether ANuA isotypes represent a diagnostic tool to predict SLE activity and define subsets of patients with LN.

Analysis of the characteristics of patients with false-positive HIV screening assay results.

OBJECTIVE: To investigate the clinical characteristics of patients with false-positive human immunodeficiency virus (HIV) screening test results and provide a theoretical basis to interpret the results of HIV screening tests. METHODS: This retrospective study evaluated the incidence of false-positive results for HIV screening tests and characterized false-positive reactions during HIV screening in a large-scale study at Zhongshan Hospital, Xiamen University. RESULTS: False-positive HIV test results occurred for 264 of 275,263 (0.10%) serum samples. Although the incidence of a false-positive HIV screening result did not differ between male and female patients (screening χ2 = 1.194; P = 0.275), it increased with age (χ2 = 25.759; P < 0.01). False-positive reactions were associated with 16 disease categories, including 101 diseases, some of which have never been reported previously to be associated with false-positive HIV screening results. CONCLUSIONS: The occurrence of false-positive HIV screening test results may indicate underlying serious disorders. Characterization of patients with false-positive HIV screening test results can help identify potential diseases unrelated to HIV.

Chemokine CXCL1 as a potential marker of disease activity in systemic lupus erythematosus.

OBJECTIVES: The chemokine CXCL1, known as growth-related oncogene α (GRO-α), is a potent chemoattractant and regulator of neutrophils. The purpose of our study was to evaluate the regulatory response of CXCL1 in the serum of patients with systemic lupus erythematosus (SLE) in the active stage of disease and to assess whether it was implicated in the pathogenesis/inflammatory process in lupus. METHODS: CXCL1 serum concentrations were examined in 90 SLE patients, 56 other autoimmune diseases (OADs) patients and 100 healthy controls using enzyme-linked immunosorbent methodology. RESULTS: SLE patients exhibited significant increases in serum CXCL1 concentrations [1492.86 (735.47-2887.34) pg/ml] compared with OADs patients [155.88 (10.77-366.78) pg/ml] and healthy controls [13.58 (8.46-37.22) pg/ml] (p < 0.001). Moreover, the level of CXCL1 decreased as the level of anti-dsDNA IgG decreased after treatment between the anti-dsDNA-positive SLE patients and the anti-dsDNA-negative SLE patients. Additionly, serum CXCL1 concentrations were related to different disease activity levels in SLE and lupus nephritis (LN) and high avidity of IgG ANAs (HA IgG ANAs) (p < 0.05). Furthermore, CXCL1 serum concentrations were significantly correlated with the SLE Disease Activity Index(SLEDAI) score, relative avidity index (RAI) of HA IgG ANAs and the levels of anti-dsDNA IgG, CRP, ESR, albumin, C3 and C4.Additionally, Statistical analysis revealed that positivity for IgG ANA (p < 0.001), the presence of HA IgG ANAs (p = 0.001) and the logarithmic level of anti-dsDNA IgG (p = 0.021) were significantly associated with the logarithmic level of CXCL1 with standard partial regression coefficients (95% CI) of 2.371 (1.734-3.009), 1.231 (0.52-1.937) and 0.409 (0.062-0.755), respectively. Finally, using cutoff points of 1182.17 pg/mL and 1500.31 pg/mL, serum CXCL1 levels had a similar sensitivity of 76% and specificity of 100% and 75% for the diagnosis of active SLE and LN, respectively. CONCLUSIONS: Serum CXCL13 concentrations might represent a potential marker of disease activity in systemic lupus erythematosus.

MiR-26a-5p Targets WNT5A to Protect Cardiomyocytes from Injury Due to Hypoxia/Reoxygenation Through the Wnt/ß-catenin Signaling Pathway.

This study aimed to investigate the effect and mechanism of miR-26a-5p on cardiomyocyte injury induced by hypoxia/reoxygenation (H/R).After construction of an H/R model in rat cardiomyocyte H9c2 cells, miR-26a-5p in the cells was interfered with (cells transfected with miR-26a-5p inhibitor) or overexpressed (cells transfected with a miR-26a-5p mimics). The viability and the apoptosis rate of cells in each group were detected using CCK-8 and flow cytometry; the relationship between miR-26a-5p and WNT5A was verified by a dual-luciferase reporter assay; the expression of miR-26a-5p, WNT5A, cleavedcaspase3 and Wnt/ß-catenin signaling pathway-related proteins in each group was detected using qRT-PCR or Western blot; LDH release, SOD, and GSH-PX activities in each group were detected by kit.In the H/R group, the expression level of miR-26a-5p was significantly decreased, whereas the expression level of WNT5A was significantly increased. The activity of the Wnt/ß-catenin signaling pathway was up-regulated; the level of LDH released was significantly increased; and activities of SOD and GSH-PX were significantly decreased. The aforementioned changes resulted in decreased cell activity and increased apoptosis rate. The overexpression of miR-26a-5p could reduce the expression level of WNT5A, the activity of the Wnt/ß-catenin signaling pathway, and the apoptosis rate and restore the cell viability.These results suggest that miR-26a-5p can target WNT5A and thus, inhibit the Wnt/ß-catenin signaling pathway activity, inhibiting H/R-induced cardiomyocyte injury and apoptosis.

Methylation of CYP1A1 and VKORC1 promoter associated with stable dosage of warfarin in Chinese patients.

OBJECTIVE: To investigate the association between DNA methylation and the stable warfarin dose through genome-wide DNA methylation analysis and pyrosequencing assay. METHOD: This study included 161 patients and genome-wide DNA methylation analysis was used to screen potential warfarin dose-associated CpGs through Illumina Infinium HumanMethylation 450 K BeadChip; then, the pyrosequencing assay was used to further validate the association between the stable warfarin dose and alterations in the methylation of the screened CpGs. GenomeStudio Software and R were used to analyze the differentially methylated CpGs. RESULTS: The methylation levels of CpGs surrounding the xenobiotic response element (XRE) within the CYP1A1 promoter, differed significantly between the different dose groups (P < 0.05), and these CpGs presented a positive correlation (r> 0, P < 0.05) with an increase in the stable dose of warfarin. At the VKORC1 promoter, two CpGs methylation levels were significantly different between the differential dose groups (P < 0.05), and one CpG (Chr16: 31106793) presented a significant negative correlation (r <  0, P <  0.05) among different dose (low, medium, and high) groups. CONCLUSION: This is a novel report of the methylation levels of six CpGs surrounding the XRE within the CYP1A1 promoter and one differential CpG at the VKORC1 promoter associated with stable warfarin dosage; these methylation levels might be applied as molecular signatures for warfarin.

MicroRNA-296-5p inhibits cell proliferation by targeting HMGA1 in colorectal cancer.

An increasing body of evidence indicates the involvement of microRNAs (miRNAs/miRs) in the initiation and progression of colorectal cancer (CRC). miR-296-5p was recently identified as a tumor suppressor in a variety of human cancer types; however, its function in CRC remains largely unknown. The present study demonstrated that the expression of miR-296-5p was significantly downregulated in CRC tissues and cell lines. The overexpression of miR-296-5p markedly inhibited proliferation, and induced cell cycle arrest and apoptosis in CRC cells. Bioinformatics analysis suggested that high mobility group AT-hook 1 (HMGA1) may be a target of miR-296-5p in CRC cells. Further experiments showed that miR-296-5p bound the 3'-untranslated region of HMGA1 and decreased its expression in CRC cells. HMGA1 was overexpressed in CRC tissues and was inversely correlated with the expression of miR-296-5p. The restoration of HMGA1 significantly reversed the inhibitory effect of miR-296-5p on the proliferation of CRC cells. Overall, the findings of the present study indicate that miR-296-5p suppressed the progression of CRC, at least partially via targeting HMGA1. Thus, miR-296-5p is a potential target for novel therapies in CRC.

Evaluation of a newly developed quantitative determination kit for tumor marker CA15-3 with chemiluminescent assay.

BACKGROUND: Tumor marker carbohydrate antigen 15-3 (CA15-3) is used as a biomarker to aid to diagnose and monitor the prognosis of breast cancer patients. A new quantitative determination kit for CA15-3 with chemiluminescent assay was developed by Xiamen InnoDx Biotech Co., Ltd, China. Therefore, we conducted the report to evaluate the performance of the kit. METHODS: According to the "Guiding principles on performance analysis of diagnostic reagents in vitro", the calibration curve, limit of detection, reportable range, accuracy, precision, anti-interference capability, cross-reaction and comparison by measuring EDTA plasma and serum were carried out. In addition, the kit was performed in parallel to electrochemiluminescence immunoassay kit (Roche) to analyze the correlation between the two kits. RESULTS: Regression equation of calibration curve of the kit was Y=0.7914X+4.1032 (R2 =.990). Limit of detection was 0.0347 U/mL. The reportable range was 0.5-2400 U/mL. Recovery ratio was 100.0%-104.8%. Coefficient of variations (CVs) of within-run and between-run were 4.8%-7.6% and 5.8%-7.4% respectively. No remarkable interferences (all Bias% were less than ±10%) were detected when samples contained hemoglobin ≤183.8 µmol/L, bilirubin ≤340 µmol/L, triglyceride ≤18.1 mmol/L, or rheumatoid factor ≤400 U/mL. No cross-reaction was present in the kit. Moreover, compared with the results from electrochemiluminescence immunoassay kit (Roche) in 345 serum samples, there was a satisfied correlation coefficient of 0.977 (P<.01), and the kit was simultaneously fit for the detection of EDTA plasma and serum samples. CONCLUSION: The new kit validated satisfactorily, and it can be used for detecting CA15-3 in clinical practice.

Diagnostic utility of the Elecsys anti-CCP assay in patients with rheumatoid arthritis.

OBJECTIVE: The automatic anti-cyclic citrullinated peptide (anti-CCP) antibodies assay offered great advantages over traditional methods in terms of improved precision, reliability, technical simplicity, short turnaround time and high-speed throughput. In this study, we evaluated the main technical performance and diagnostic accuracy of the first automatic anti-CCP assay approved in China. METHODS: The study comprised 106 rheumatoid arthritis (RA) patients, 203 non-RA rheumatic disease controls and 46 healthy persons. Anti-CCP, rheumatoid factor (RF), α1-acid glycoprotein, C-reactive protein and erythrocyte sedimentation rate were measured and compared. The precision, reference intervals for Chinese population and cut-off value for RA diagnosis, as well as the suitable diluent for anti-CCP were assessed. The positive rate and score of anti-CCP were compared with RF and acute-phase reactants, according to the new RA criteria. RESULTS: Within- and between-run imprecision, expressed as the coefficient of variation, were 0.47-1.36% and 1.15-2.63%, respectively. Upper 95% reference limit of anti-CCP in healthy Chinese was 8.8 U/mL. The area under curve of the receiver operating characteristic(ROC) for anti-CCP and RF were 0.882 (95% CI 0.833-0.930) and 0.844 (95% CI 0.792-0.897), respectively. Based on the cut-off value set by ROC, compared to RF, anti-CCP had higher sensitivity (96.8% vs. 78.3%) and specificity (90.9% vs. 70.7%). With 17 U/mL set as the optimal cut-off for anti-CCP, the total positivity of anti-CCP was comparable to that of RF (76.4% vs. 75.5%), but the high-positivity rate of anti-CCP was significantly higher (74.5% vs. 62.3%, p < 0.005). CONCLUSIONS: Our results confirm anti-CCP as a more sensitive and specific marker than RF for the diagnosis of RA. The diagnostic performance of the Elecsys anti-CCP assay makes it a useful adjunct to clinical practice in the Chinese population.