Human Epididymis Protein Research Articles

Recent Insight about HE4 Role in Ovarian Cancer Oncogenesis.

Currently, ovarian cancer (OC) is a target of intense biomarkers research because of its frequent late diagnosis and poor prognosis. Serum determination of Human epididymis protein 4 (HE4) is a very important early detection test. Most interestingly, HE4 plays a unique role in OC as it has been implicated not only in OC diagnosis but also in the prognosis and recurrence of this lethal neoplasm, actually acting as a clinical biomarker. There are several evidence about the predictive power of HE4 clinically, conversely less has been described concerning its role in OC oncogenesis. Based on these considerations, the main goal of this review is to clarify the role of HE4 in OC proliferation, angiogenesis, metastatization, immune response and also in the development of targeted therapy. Through a deeper understanding of its functions as a key molecule in the oncogenetic processes underlying OC, HE4 could be possibly considered as an essential resource not only for diagnosis but also for prognosis and therapy choice.

Association between serum HE4 and poor periodontal health in adult women

ObjectivesThe aim of this study is to explore the association between serum human epididymal protein (HE4) levels and poor periodontal health.Materials and methodsData used in our study from the National Health and Nutrition Examination Survey (NHANES) 2001–2002 and Gene Expression Omnibus database (GSE10334 and GSE16134). Periodontitis category was defined by the 2017 classification scheme based on clinical periodontal parameters. Univariate and multivariate logistic regression analyses were used to explore the relationship between serum HE4 levels and the risk of periodontitis. GSEA analysis was performed to investigate the function of HE4.ResultsA total of 1715 adult women over the age of 30 were included in our study. Compared with the lowest tertile, individuals in the highest tertile of HE4 levels were more likely to be Stage III/IV periodontitis (ORtertile3vs1 = 2.35, 95% CI: 1.35–4.21). The association was still significant in populations who were less than 60 years old, non-Hispanic white, high school graduate, 1.3 < PI ≤ 3.5, non-smoker, current smoker, non-obese, obese, and who had not diabetes mellitus or had not hypertension. In addition, HE4 expression was upregulated in diseased gingival tissues and involved in cell proliferation and immunity.ConclusionsSerum HE4 is positively associated with poor periodontal health in adult women.Clinical relevancePatients with high serum HE4 levels are more likely to have Stage III/IV periodontitis. HE4 has the potential to be used as a biomarker to predict the severity of periodontitis.

Diagnostic performance of IOTA SR and O-RADS combined with CA125, HE4, and risk of malignancy algorithm to distinguish benign and malignant adnexal masses

PurposeTo compare the diagnostic performance of International Ovarian Tumour Analysis Simple Rules (IOTA SR) and Ovarian-Adnexal Reporting and Data System (O-RADS), and to analyse whether combining IOTA SR and O-RADS with the biomarkers cancer antigen 125 (CA125), human epididymis protein 4 (HE4), and risk of malignancy algorithm (ROMA) further improves diagnostic performance in women with different menopause status. MethodsThis study retrospectively included patients with ovarian adnexal masses confirmed by surgical pathology between September 2021 and February 2022. The area under the curve (AUC), sensitivity, and specificity were calculated to evaluate the diagnostic efficacy of IOTA SR, O-RADS, and their combination with CA125, HE4, and ROMA. ResultsThis study included 1,179 ovarian adnexal masses. In all women, the AUC of IOTA SR was comparable to O-RADS (0.879 vs. 0.889, P = 0.361), and O-RADS had a significantly higher sensitivity than IOTA SR (95.77 % vs. 87.32 %, P < 0.001). In premenopausal women, O-RADS had a significantly higher AUC than other diagnostic strategies (all P < 0.05), and the sensitivity, specificity, and accuracy were 93.33 %, 84.74 %, and 85.59 %, respectively. In postmenopausal women, IOTA SR + ROMA had a significantly higher AUC than other diagnostic strategies (all P < 0.05), and the sensitivity, specificity, and accuracy were 85.37 %, 93.88 %, and 90.00 %, respectively. ConclusionsOur study supports the high diagnostic value of IOTA SR or O-RADS alone in all women, and O-RADS was more sensitive than IOTA SR. In premenopausal women, O-RADS had the highest diagnostic value. In postmenopausal women, IOTA SR outperformed O-RADS, and IOTA SR + ROMA had the highest diagnostic value.

Investigation of Serum Human Epididymitis Protein 4 Level in Rats with Experimental Acute Pancreatitis.

We aimed to show whether the serum level of Human Epididymitis Protein 4 increases in rats with an experimental acute pancreatitis model created by cerulein. This study included 24 male Sprague-Dawley rats which were randomly divided into 4 groups each containing 6 rats. the group treated with saline, Group 1: pancreatitis group created with cerulein at a total dose of 80 µg/kg, Group 2: pancreatitis group created with cerulein at a total dose of 120 µg/kg, Group 3: pancreatitis group created with cerulein at a total dose of 160 µg/kg. There were statistically significant differences between edema, acinar necrosis, fat necrosis, and perivascular inflammation scores among the study groups. While the degree of all histopathological findings is lowest in the control group, pancreatic parenchyma damage increases as the amount of injected cerulein increases. There was no statistically significant difference between alanine aminotransferase, aspartate aminotransferase, and Human Epididymis Protein 4 values between study groups. On the other hand, there was a statistically significant difference between amylase and lipase values. The lipase value of the control group was significantly lower than the lipase value of the second and third groups. The amylase value of the control group was significantly lower than all other groups. The highest Human Epididymis Protein 4 value was measured as 104 pmol/L in the first pancreatitis group, where the severity of pancreatitis was mild. In the present study, it was concluded that the Human Epididymis Protein 4 value increased in the case of mild pancreatitis, but there is no correlation between the severity of pancreatitis and the Human Epididymis Protein 4 value.

P174 Human epididymis protein 4 (HE4) plasma concentrations correlate with the improvement of ppFEV1 in response to LUM/IVA therapy in people with cystic fibrosis homozygous for p.Phe508del-CFTR

We have previously documented that elevated plasma HE4 concentrations decreased in people with CF (pwCF) bearing p.Gly551Asp-CFTR mutation in response to ivacaftor (IVA), and these concentrations were inversely correlated with ppFEV1. Although the PROSPECT study has evaluated the effectiveness of lumacaftor (LUM)/IVA in pwCF homozygous for p.Phe508del-CFTR variant, plasma biomarkers have not been used to monitor treatment efficacy.

A signal-on electrochemical aptasensor for sensitive detection of human epididymis protein 4 based on functionalized metal–organic framework/ketjen black nanocomposite

A signal-on electrochemical aptasensor for sensitive detection of human epididymis protein 4 based on functionalized metal–organic framework/ketjen black nanocomposite

Retrospective efficacy analysis of olaparib combined with bevacizumab in the treatment of advanced colorectal cancer

Colorectal cancer (CRC) is a highly prevalent malignancy of the digestive tract worldwide, characterized by a significant morbidity and mortality rate and subtle initial symptoms. Diarrhea, local abdominal pain, and hematochezia occur with the development of cancer, while systemic symptoms such as anemia and weight loss occur in patients with advanced CRC. Without timely interventions, the disease can have fatal consequences within a short span. The current therapeutic options for colon cancer include olaparib and bevacizumab, which are widely utilized. This study intends to evaluate the clinical efficacy of olaparib combined with bevacizumab in the treatment of advanced CRC, hoping to provide insights into advanced CRC treatment.To investigate the retrospective efficacy of olaparib combined with bevacizumab in the treatment of advanced CRC.A retrospective analysis was conducted on a cohort of 82 patients with advanced colon cancer who were admitted to the First Affiliated Hospital of the University of South China between January 2018 and October 2019. Among them, 43 patients subjected to the classical FOLFOX chemotherapy regimen were selected as the control group, and 39 patients undergoing treatment with olaparib combined with bevacizumab were selected as the observation group. Subsequent to different treatment regimens, the short-term efficacy, time to progression (TTP), and incidence rate of adverse reactions between the two groups were compared. Changes in serum-related indicators [vascular endothelial growth factor (VEGF), matrix metalloprotein-9 (MMP-9), cyclooxygenase-2 (COX-2)] and tumor markers [human epididymis protein 4 (HE4), carbohydrate antigen 125 (CA125), carbohydrate antigen 199 (CA199)] levels before and after treatment were compared between the two groups at the same time.The objective response rate was discovered to be 82.05%, and the disease control rate was 97.44% in the observation group, which were significantly higher than the respective rates of 58.14% and 83.72% in the control group (P < 0.05). The median TTP was 24 mo (95%CI: 19.987-28.005) in the control group and 37 mo (95%CI: 30.854-43.870) in the observation group. The TTP in the observation group was significantly better than that in the control group, and the difference held statistical significance (log-rank test value = 5.009, P = 0.025). Before treatment, no substantial difference was detected in serum VEGF, MMP-9, and COX-2 levels and tumor markers HE4, CA125, and CA199 levels between the two groups (P > 0.05). Following treatment with different regimens, the above indicators in the two groups were remarkably promoted (P < 0.05), VEGF, MMP-9, and COX-2 in the observation group were lower than those in the control group (P < 0.05), and HE4, CA125, and CA199 levels were also lower than those in the control group (P < 0.05). Vis-à-vis the control group, the total incidence of gastrointestinal reactions, thrombosis, bone marrow suppression, liver and kidney function injury, and other adverse reactions in the observation group was notably lowered, with the difference considered statistically significant (P < 0.05).Olaparib combined with bevacizumab in the treatment of advanced CRC demonstrates a strong clinical effect of delaying disease progression and reducing the serum levels of VEGF, MMP-9, COX-2 and tumor markers HE4, CA125 and CA199. Moreover, given its fewer adverse reactions, it can be regarded as a safe and reliable treatment option.

Elevated serum levels of human epididymis protein 4 in adult patients with proliferative lupus nephritis.

This study aimed to access whether serum human epididymis protein 4 (HE4) level could identify lupus nephritis (LN) pathological classes in adults and children. The serum HE4 levels of 190 healthy subjects and 182 patients with systemic lupus erythematosus (SLE) (61 adult-onset LN [aLN], 39 childhood-onset LN [cLN], and 82 SLE without LN) were determined using Architect HE4 kits and an Abbott ARCHITECT i2000SR Immunoassay Analyzer. Serum HE4 level was significantly higher in the aLN patients (median, 85.5 pmol/L) than in the patients with cLN (44 pmol/L, P < 0.001) or SLE without LN (37 pmol/L, P < 0.001), or the healthy controls (30 pmol/L, P < 0.001). Multivariate analysis showed that serum HE4 level was independently associated with aLN. Stratified by LN class, serum HE4 level was significantly higher in the patients with proliferative LN (PLN) than in those with non-PLN, and this difference was found only in aLN (median, 98.3 versus 49.3 pmol/L, P = 0.021) but not in cLN. Stratified by activity (A) and chronicity (C) indices, the aLN patients with class IV (A/C) possessed significantly higher serum HE4 levels than those with class IV (A) (median, 195.5 versus 60.8 pmol/L, P = 0.006), and this difference was not seen in the class III aLN or cLN patients. Serum HE4 level is elevated in patients with class IV (A/C) aLN. The role of HE4 in the pathogenesis of chronic lesions of class IV aLN needs further investigation.

Endometrioid adenocarcinoma: combined multiparametric MRI and tumour marker HE4 to evaluate tumour grade and lymphovascular space invasion

To assess the value of semi-quantitative dynamic contrast-enhanced magnetic resonance imaging (DCE-MRI) and quantitative diffusion-weighted imaging parameters combined with human epididymis protein 4 (HE4) in predicting the pathological grade and lymphovascular space invasion (LVSI) of endometrioid adenocarcinoma (EAC). Between October 2018 and December 2021, 60 women (mean age, 55 [range, 32-77] years) with EAC underwent preoperative pelvic MRI and HE4 level measurements. The positive enhancement integral (PEI), time to peak, maximum slope of increase (MSI), and maximum slope of decrease were measured by manually drawing a region of interest on the neoplastic tissue. The receiver operating characteristic curve was used to calculate the diagnostic efficiency of the single parameter and combined factors. Lower apparent diffusion coefficients (ADCs) were observed in high-grade tumours (G3) than in low-grade tumours (G1/G2). PEI, MSI, and HE4 levels were higher in the high-grade tumours than in the low-grade tumours (p<0.05). The area under the curve (AUC) for G3 diagnosis using multiparametric MRI combined with HE4 was 0.929. ADC values were significantly lower in the EAC with LVSI than in those without LVSI. Tumours with LVSI showed higher PEI and HE4 levels than those without LVSI (p<0.05). The AUC for LVSI-positive diagnosis using multiparametric MRI combined with HE4 was 0.814. Semi-quantitative DCE-MRI, ADC values, and serum HE4 levels can be used to predict tumour grade and LVSI, and the prediction efficiency of multiparametric MRI combined with serum HE4 is better than that of any single factor.

Human epididymis protein 4 (HE4) plasma concentration inversely correlates with the improvement of cystic fibrosis lung disease in p.Phe508del-CFTR homozygous cases treated with the CFTR modulator lumacaftor/ivacaftor combination

BackgroundWe previously documented that elevated HE4 plasma concentration decreased in people with CF (pwCF) bearing the p.Gly551Asp-CFTR variant in response to CFTR modulator (CFTRm) ivacaftor (IVA), and this level was inversely correlated with the FEV1% predicted values (ppFEV1). Although the effectiveness of lumacaftor (LUM)/IVA in pwCF homozygous for the p.Phe508del-CFTR variant has been evaluated, plasma biomarkers were not used to monitor treatment efficacy thus far. MethodsPlasma HE4 concentration was examined in 68 pwCF drawn from the PROSPECT study who were homozygous for the p.Phe508del-CFTR variant before treatment and at 1, 3, 6 and 12 months after administration of LUM/IVA therapy. Plasma HE4 was correlated with ppFEV1 using their absolute and delta values. The discriminatory power of delta HE4 was evaluated for the detection of lung function improvements based on ROC-AUC analysis and multiple regression test. ResultsHE4 plasma concentration was significantly reduced below baseline following LUM/IVA administration during the entire study period. The mean change of ppFEV1 was 2.6% (95% CI, 0.6 to 4.5) by 6 months of therapy in this sub-cohort. A significant inverse correlation between delta values of HE4 and ppFEV1 was observed especially in children with CF (r=-0.7053; p<0.0001). Delta HE4 predicted a 2.6% mean change in ppFEV1 (AUC: 0.7898 [95% CI 0.6823–0.8972]; P < 0.0001) at a cut-off value of -10.7 pmol/L. Moreover, delta HE4 independently represented the likelihood of being a responder with ≥ 5% delta ppFEV1 at 6 months (OR: 0.89, 95% CI: 0.82–0.95; P = 0.001). ConclusionsPlasma HE4 level negatively correlates with lung function improvement assessed by ppFEV1 in pwCF undergoing LUM/IVA CFTRm treatment.

Is Serum Human Epididymis Protein HE4, More Specific than Cancer Antigen 125 in Patients with Benign Gynecological Diseases?

Background: The cancer antigen 125 “CA125” and Human Epididymis protein 4 “HE4” are among some well worked out markers for recognition of malignancy in ovarian tumors. Their different cutoffs are determined and reported by various studies and mostly used recommended cutoffs are 35.0 U/mL for CA125 and 140pmol/L for HE4. They are both reported to recognize the malignancy well, but how specific they are for benign gynecological diseases is still needed to be worked out. Aim: To determine and compare the performance of CA125 and HE4 for benign gynecological pathologies. Methodology: A cross-sectional study was performed at Gajju Khan Medical College in collaboration with Armed forces Institute of pathology (AFIP). A total of 76 women of age above 18 years age, with benign ovarian pathology, confirmed through histological evaluation were enrolled. All these were examined on both markers, i.e. CA125 and HE4. Standard cutoffs of CA125 &gt;35.0 U/mL and HE4&gt;140 pmol/L were used to see if the case was labeled malignant, incorrectly. SPSS 20.0 was used to manage and analyze the data. Performance of the two markers were compared by using some descriptive analysis and using chi-square test. P-value ≤0.05 was considered significant. Results: The median age of these women was 40 years ranging between 18 and 71. Majority (86.8%) was premenopausal, The CA125 had no difference between pre and post-menopausal women but HE4 had a significant difference with p-value 0.029. there were 10.5% incorrect labeling of malignancy when evaluated for CA125, while only 2.6% incorrect on HE4. This difference was found statistically significant between two markers with p-value 0.05. Conclusion: HE4 is better and more specific marker for benign gynecological pathologies as compared to CA125 Keywords:Benign Ovarian Epithelial, Human epididymisprotein (HE4), Cancer Antigen 125 (CA125), Specificity

Nomogram to predict the incidence of new-onset heart failure after acute coronary syndrome among women.

Although great progress has been made in caring for patients with acute coronary syndrome (ACS), the incidence of heart failure (HF) after discharge remains high after ACS. We aimed to investigate the risk predictors for new-onset HF and build a simple nomogram to optimize the clinical management of female patients. The clinical data of 319 female patients with ACS between January 1, 2021 and January 1, 2022, were obtained from the Zhejiang Provincial People's Hospital. Multivariate logistic regression analysis was carried out to build the prediction model among all participants and then verified by 10-fold cross-validation. The discrimination, calibration, and clinical usefulness of the prediction model were assessed using receiver operating characteristic curve, calibration curve, and decision curve analyses. This study analyzed 15 potential independent risk predictors of new-onset HF in 319 female patients with ACS. The incidence of HF onset was 23.2%. The following 5 independent risk predictors were filtered out as most relevant for predicting 12-month HF onset: left ventricular ejection fraction ≤ 60.5%, high-density lipoprotein ≤ 1.055 mmol/L, human epididymal protein 4 > 69.6 pmol/L, creatinine > 71.95 µmol/L, and diagnosis of myocardial infarction (MI). Our nomogram, which used five easily obtained clinical variables, could be a useful tool to help identify female individuals with ACS who are at high risk of developing HF after discharge and facilitate communication between female patients and physicians.

Nanopore-Based Fingerprint Immunoassay Based on Rolling Circle Amplification and DNA Fragmentation.

In recent years, nanopore-based sequencers have become robust tools with unique advantages for genomics applications. However, progress toward applying nanopores as highly sensitive, quantitative diagnostic tools has been impeded by several challenges. One major limitation is the insufficient sensitivity of nanopores in detecting disease biomarkers, which are typically present at pM or lower concentrations in biological fluids, while a second limitation is the general absence of unique nanopore signals for different analytes. To bridge this gap, we have developed a strategy for nanopore-based biomarker detection that utilizes immunocapture, isothermal rolling circle amplification, and sequence-specific fragmentation of the product to release multiple DNA reporter molecules for nanopore detection. These DNA fragment reporters produce sets of nanopore signals that form distinctive fingerprints, or clusters. This fingerprint signature therefore allows the identification and quantification of biomarker analytes. As a proof of concept, we quantify human epididymis protein 4 (HE4) at low pM levels in a few hours. Future improvement of this method by integration with a nanopore array and microfluidics-based chemistry can further reduce the limit of detection, allow multiplexed biomarker detection, and further reduce the footprint and cost of existing laboratory and point-of-care devices.

Label-free electrochemical bioplatform based on Au-modified magnetic Fe3O4/α-Fe2O3 hetero-nanorods for sensitive quantification of ovarian cancer tumor marker

Due to challenges in the ultrasensitive detection of human epididymis protein 4 (HE4), a vital tumor marker for the early diagnosis of ovarian cancer (OC), it is necessary to establish efficient methods for HE4 determination. Recently, electrochemical bioplatforms combined with nanomaterials have shown outstanding performance in the analysis of biological molecules. The surface modification of conductive materials is essential for signal amplification and detection sensitivity. Herein, we developed a novel label-free HE4 electrochemical bioplatform by introducing magnetic Fe3O4/α-Fe2O3 hetero-nanorods (Fe3O4/α-Fe2O3 NRs) as a substrate for signal amplification and DNA aptamer (Apt) probe immobilization in combination with magnetic self-assembly and magnetic separation. Differential pulse voltammetry (DPV) results verified that the platform had excellent specificity for HE4 under optimized conditions and was positively linearly correlated with the logarithm of HE4 concentration (R2 = 0.996) with a linear range of 0.1 pg mL−1 to 1 ng mL−1 and detection limit (LOD) of 27.5 fg mL−1. Moreover, the bioplatform demonstrated promising results for the detection of HE4 in spiked serum samples.

Sensitive Detection of the Human Epididymis Protein-4 (HE4) Ovarian Cancer Biomarker through a Sandwich-Type Immunoassay Method with Laser-Induced Breakdown Spectroscopy.

Detection of cancer before the appearance of any symptoms is crucial for successful treatment. Early detection is, however, very challenging, particularly for the types of cancer with few or no symptoms at early stages, such as epithelial ovarian cancer (EOC). Developing a user-friendly method that can detect biomarkers with sufficient selectivity, sensitivity, and reproducibility is a promising approach for overcoming the challenges of early detection of EOC. In this study, we report a sandwich-type microparticle immunoassay for sensitive detection of the HE4 biomarker with laser-induced breakdown spectroscopy. Here, we cross-linked elemental particles to a specific functional group of the targeted biomolecules based on a covalent and non-covalent linking chemistry to improve the sensitivity and selectivity of biomarker detection, in which Fe3O4 and SiO2 microparticles were used to conjugate and purify the antibody-antigen in complex media. Simultaneous detection of Fe and Si from a magnetically purified assay significantly improves the HE4 biomarker's detectability, in which HE4 was detected with a limit of detection of 0.0022 pM. We also determined the coupling ratio between HE4 and silica particles using a silicon calibration curve.

Prognostic Role of Human Epididymis Protein4 (HE4) in Endometrial Lesions: Study in a Tertiary Care Centre.

Endometrial cancer accounts for 20% of malignant tumours in the female reproductive system. A novel biological marker human epididymis protein4 (HE4) represents an important alternative indicator which may benefit patient mortality. To correlate the immunohistochemical expression of HE4 in different non-neoplastic and neoplastic endometrial lesions and with the WHO grade of the tumours. Our study was a cross-sectional, observational study done in a tertiary care hospital from December 2019 to June 2021 on the hysterectomy sample of 50 patients with a clinical history of abnormal uterine bleeding and pelvic pain. The study showed strong positivity of HE4 in cases of endometrial carcinoma, weak positivity in cases of atypical endometrial hyperplasia, and negativity in cases of endometrial hyperplasia without atypia group. WHO grade 3(50%) and grade 2 (29%) endometrioid adenocarcinoma NOS in our study showed strong positivity for HE4 which was statistically significant (P value = 0.001). In recent studies using overexpression of HE4-related genes, the malignant biological behaviour such as cell adhesion, invasion, and proliferation was enhanced. It was seen in our study that strong positivity of HE4 was seen in all endometrial carcinoma groups and with higher WHO grade. So, HE4 may become a potential therapeutic target for advanced-stage endometrial carcinoma which requires further research. Thus, human epididymis-specific protein 4 (HE4) has been shown to be a promising marker for the detection of endometrial carcinoma patients who could be benefitted from targeted therapy.

Human epididymis protein 4 as a clinical biomarker in identifying interstitial lung disease in patients with idiopathic inflammatory myopathies.

Human epididymis protein 4 (HE4) can differentiate interstitial lung disease from patients with some rheumatic diseases. However, the clinical utility of HE4 in idiopathic inflammatory myopathies (IIM) remains unclear. 80 IIM patients and 91 age and gender-matched healthy controls (HCs) were recruited. Clinical and laboratory data were recorded at baseline and 12weeks. HE4 was tested by the method of electrochemical luminescence. Compared to HCs, the levels of HE4 significantly elevated in IIM patients. Patients with elevated HE4 had a higher interstitial lung disease (ILD) prevalence. Among patients with ILD, histological patterns of organizing pneumonia had higher HE4 levels than non-specific interstitial pneumonia. Further, there was a positive correlation between HE4 and the semi-quantitative CT grade (r=0.778, p<0.001) and a negative relation between HE4 and the percentage of forced vital capacity (p<0.001) and diffusing capacity of the lung for carbon monoxide (DLco) (p=0.001). An optimal cut-off value of HE4 (79.6pmol/L) for distinguishing IIM-ILD was analyzed by ROC analysis with an AUC of 0.733 (p=0.002). Regression analysis revealed that elevated HE4 independently identified IIM-related ILD (OR 34.8, 95%CI, 3.58-338.14, p=0.002). With the improvement after treatment, serum HE4 levels were significantly decreased (p=0.006), accompanied by improved DLco% (p=0.012). Serum HE4 was significantly elevated in patients with IIM and may be utilized as a serum biomarker to evaluate the disease severity and prognosis of IIM-related ILD.

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

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. 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. 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. 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 takingbothCKD and ovarian cancer into consideration.

Establishment and validation of reference intervals for tumor markers (AFP, CEA, CA19-9, CA15-3, CA125, PSA, HE4, Cyfra 21-1, and ProGRP) in primary care centers in Korea: A cross-sectional retrospective study.

The reference interval (RI) for a tumor marker may vary between populations, detection systems, and the methods used to obtain their values. The aims of this study were to establish age- and sex-specific RIs for the following nine common tumor markers and to validate the established RIs in Korean adults: alpha fetoprotein (AFP), carcinoembryonic antigen (CEA), cancer antigen (CA)19-9, CA15-3, CA125, Human epididymis protein 4 (HE4), total prostate specific antigen, cytokeratin fragment (Cyfra) 21-1, and progastrin-releasing peptide (ProGRP). This cross-sectional study consecutively selected 214,159 individuals (aged 18-98 years) who underwent health checkups at 16 health-promotion centers in 13 Korean cities. Finally, 62,752 examinees were used to establish the RIs after removing outliers. RIs were established using an indirect method according to the CLSI EP28-A3C guideline. The established RIs were validated by calculating the proportion of individuals outside each RI. Sex-related differences were observed for AFP, CEA, CA19-9, Cyfra 21-1, and ProGRP (p < 0.05): AFP, CEA and Cyfra 21-1 were higher in males, and CA19-9 and proGRP were higher in females. Most of the tumor markers except CA15-3 and CA125 increased with age: CA125 decreased at ≥50 years of age (p < 0.05), while CA15-3 did not vary with age. Less than 5% of subjects were outside all RIs (the 2.5th and 97.5th percentiles) established in the present study. Meanwhile, less than 3% of the healthy reference subjects fell outside the current and manufacturers' RIs of all tumor markers except Cyfra 21-1. This study has determined age- and sex-specific RIs for nine common tumor markers in the healthy Korean population, which could be useful for clinicians making clinical decisions and assessments.

Nanopore-based immunoassay based on rolling circle amplification and DNA fragmentations.

In recent years, small-size portable nanopore-based sequencers have become a robust tool with unique advantages for genomics applications. However, a few challenges have impeded applying nanopore sequencers as sensitive, quantitative diagnostic tools. One of the major bottlenecks is the inefficient capture of molecules into a pore at pM or lower concentrations which are typical for disease biomarkers. To bridge this gap we have developed a signal amplification strategy utilizing immunocapture, isothermal rolling circle amplification, and sequence-specific fragmentation of the product to release multiple DNA reporter molecules for nanopore detection. This group of DNA reporter molecules that comprise different lengths and structures could produce a distinct nanopore cluster spectrum in the two-parameter space of dwell times and fractional current blockades. By identifying the nanopore cluster spectrum, we could identify the biomarker analytes and achieve quantification. As a proof of concept, we demonstrate the quantification of human epididymis protein 4 (HE4) to low pM levels using wild-type alpha-hemolysin. Future improvement that employ microfluidics and nanopore arrays can further reduce the limit of detection, allow multiplexed biomarker detection, and further, reduce the footprint, cost, and portability of existing laboratory and point-of-care devices.