A Novel Scoring System for Early Assessment of the Risk of the COVID-19-associated Mortality in Hospitalized Patients: COVID-19 BURDEN

Background: Corona Virus Disease 2019 (COVID-19) presentation resembles common flu or can be more severe; it can result in hospitalization with significant morbidity and/or mortality. We made an attempt to develop a predictive model and a scoring system to improve the diagnostic efficiency for COVID-19 mortality via analysis of clinical features and laboratory data on admission. Methods: We retrospectively enrolled 480 consecutive adult patients, aged 21-95, who were admitted to Faghihi Teaching Hospital. Clinical and laboratory features were extracted from the medical records and analyzed using multiple logistic regression analysis. Results: A novel mortality risk score (COVID-19 BURDEN) was calculated, incorporating risk factors from this cohort. CRP (> 73.1 mg/L), O2 saturation variation (greater than 90%, 84-90%, and less than 84%), increased PT (>16.2s), diastolic blood pressure (≤75 mmHg), BUN (>23 mg/dL), and raised LDH (>731 U/L) are the features comprising the scoring system. The patients are triaged to the groups of low- (score <4) and high-risk (score ≥ 4) groups. The area under the curve, sensitivity, and specificity for predicting non-response to medical therapy with scores of ≥ 4 were 0.831, 78.12%, and 70.95%, respectively. Conclusion: Using this scoring system in COVID-19 patients, the severity of the disease will be determined in the early stages of the disease, which will help to reduce hospital care costs and improve its quality and outcome.

Serum exosomal miR-451a acts as a candidate marker for pancreatic cancer

Objectives The aim of this study was to explore the diagnostic efficiency of serum exosomal miR-451a as a novel biomarker for pancreatic cancer. Methods Serum samples were collected prior to treatment. First, we analyzed microRNA (miRNA) profiles in serum exosomes from eight pancreatic cancer patients and eight healthy volunteers. We then validated the usefulness of the selected exosomal miRNAs as biomarkers in another 191 pancreatic cancer patients, 95 pancreatic benign disease (PB) patients, and 90 healthy controls. Results The expression of miR-451a in serum-derived exosomes from pancreatic cancer patients was significantly upregulated compared with those from PB patients and healthy individuals. Serum exosomal miR-451a showed excellent diagnostic power in identifying pancreatic cancer patients. In addition, exosomal miR-451a showed a significant association with clinical stage and distant metastasis in pancreatic cancer, and the expression level of serum exosomal miR-451a was sensitive to therapy and relapse. Conclusions Serum exosomal miR-451a might serve as a novel diagnostic marker for pancreatic cancer.

Identification and validation of plasma biomarkers for diagnosis of breast cancer in South Asian women

Breast cancer is the most common malignancy among women globally. Development of a reliable plasma biomarker panel might serve as a non-invasive and cost-effective means for population-based screening of the disease. Transcriptomic profiling of breast tumour, paired normal and apparently normal tissues, followed by validation of the shortlisted genes using TaqMan® Low density arrays and Quantitative real-time PCR was performed in South Asian women. Fifteen candidate protein markers and 3 candidate epigenetic markers were validated first in primary breast tumours and then in plasma samples of cases [N = 202 invasive, 16 DCIS] and controls [N = 203 healthy, 37 benign] using antibody array and methylation specific PCR. Diagnostic efficiency of single and combined markers was assessed. Combination of 6 protein markers (Adipsin, Leptin, Syndecan-1, Basic fibroblast growth factor, Interleukin 17B and Dickopff-3) resulted in 65% sensitivity and 80% specificity in detecting breast cancer. Multivariate diagnostic analysis of methylation status of SOSTDC1, DACT2, WIF1 showed 100% sensitivity and up to 91% specificity in discriminating BC from benign and controls. Hence, combination of SOSTDC1, DACT2 and WIF1 was effective in differentiating breast cancer [non-invasive and invasive] from benign diseases of the breast and healthy individuals and could help as a complementary diagnostic tool for breast cancer.

SAA4 is a Diagnostic Marker to Enhance Detection Forrheumatoid Arthritis Combined with Anti-CCP

Objective Serum amyloid A4 (SAA4) is an apolipoprotein that is associated with high-density lipoprotein (HDL) in plasma. In this present investigation, we appraised the potential of SAA4 as a novel diagnostic biomarker for rheumatoid arthritis (RA) combined with other established RA biomarkers, including anticitrullinated protein antibody (anti-CCP), rheumatoid factor (RF)，and C-reactive protein (CRP). Based on the correlative measures of the biomarkers, we developed a diagnostic model of RA by integrating serum levels of SAA4 with these clinical parameters. Methods A number of 316 patients were recruited in the current research. The serum levels of SAA4 were assessed by quantitative ELISA. The specificity and sensitivity of biomarkers were evaluated by using a receiver-operator curve (ROC) analysis to determine their diagnostic efficiency. Univariate and multivariate logistic regression analyses were used to screen and construct the diagnostic models for RA , consisting of diagnostic biomarkers and clinical data. A diagnostic nomogram was then generated based on logistic regression analysis results. Results The serum levels of SAA4 were considerably greatest in RA patients in comparison to other control subjects (P<0.001). Compared with anti-CCP, RF and CRP respectively, SAA4 had the highest specificity (88.60%) for diagnosing RA. The combination of SAA4 with anti-CCP could have the highest diagnostic accuracy when paired together, with highest sensitivity (91.14%) in parallel and highest specificity(98.10) in series. We successfully developed two diagnostic models: the combined model of SAA4 and anti-CCP (model A), and the combined model of SAA4, CRP, anti-CCP, RF and history of diabetes (model B). Both models showed a great area under the curve of ROC for either the training cohort or the validation cohort. The data indicated that the novel RA diagnostic models possessed an advantageous discrimination capacity and application potential. Conclusion Serum SAA4 has utility as a biomarker for RA’s diagnosis and can enhance the detection of RA when combined with anti-CCP.

Development of the ELISA Test System for Quantitative Determination of IgG to the Varizella zoster virus in Human Serum and Assessment of its Diagnostic Efficiency

Relevance. The introduction of Varicella vaccine prophylaxis explains the need to develop a methodology for monitoring the vaccination effectiveness and the intensity of population immunity. This problem can be solved using quantitative immunoassay methods. Aim. Development of an enzyme-linked immunosorbent assay for the concentration of class G immunoglobulins (AB) to Varicella zoster virus (VZV) determining and assessing its functional characteristics and diagnostic efficiency. Materials and methods. Recombinant antigen GE VZV. WHO International Standard for Antibodies to VZV W1044. Blood serum samples from healthy people and patients with Chickenpox and Herpes zoster, blood serum samples containing IgG antibodies to herpes simplex viruses of the first and second types, cytomegalovirus, Epstein-Barr virus. Anti-VZV ELISA (IgG) reagent kit (Euroimmun, Germany). Indirect enzyme-linked immunosorbent assay. Immunization of animals with recombinant antigen GE, isolation, and purification of specific antibodies. Conjugation of monoclonal antibodies to human IgG with antibodies to antigen GE and with horseradish peroxidase. Results. An enzyme-linked immunosorbent assay in «an indirect» format has been developed to determine the specific antibodies to VZV concentration (IU/ml) in human serum/plasma. An artificial calibrator for determining the concentration of AB-VZV had been synthesized and standardized according to the International WHO-standard W1044. The main functional characteristics of the developed enzyme-linked immunosorbent assay are determined in accordance with GOST 51352-2013. The diagnostic kit was tested on blood serum samples from children with chickenpox (n = 43), adults with Herpes zoster (n = 158), healthy individuals (n = 781). The diagnostic sensitivity of the test system was 85%, the diagnostic specificity was 87% according to the ROC analysis. The absence of cross-reactivity of the test system was shown on samples with serological markers of other herpesvirus infections (n = 94). Comparative trials of the developed test system and its commercial analog, the Anti-VZV ELISA (IgG) reagent kit, did not reveal statistically significant differences between their functional characteristics. Conclusions. The developed test system for determining of the AB-VZV concentration in human serum/plasma in terms of its functional characteristics meets the GOST requirements, is characterized by high diagnostic efficiency, can be used to monitor the effectiveness of vaccine prophylaxis and strength of population immunity, as well as to assess the immune response in chickenpox and Herpes zoster.

Value of Contrast-Enhanced Ultrasound in Adjusting the Classification of Chinese-TIRADS 4 Nodules

Objectives. To explore the value of applying contrast-enhanced ultrasound (CEUS) in adjusting the classification of category 4 nodules in the Chinese-Thyroid Imaging Report and Data System (C-TIRADS). Methods. The data of preoperative conventional ultrasound and CEUS examinations of 125 C-TIRADS 4 nodules in 109 patients were retrospectively analyzed. We divided the thyroid nodules into two groups based on whether recommend by the guide fine-needle aspiration (FNA). Group I included C-TIRADS 4A nodules with a maximum diameter ≤15 mm and C-TIRADS 4B and 4C nodules with a maximum diameter ≤10 mm, and Group II included C-TIRADS 4A nodules with a maximum diameter >15 mm and C-TIRADS 4B and 4C nodules with a maximum diameter >10 mm. In CEUS, thyroid nodules showing suspicious malignant features such as hypoenhancement or early washout were adjusted to a level higher in the C-TIRADS classification; thyroid nodules showing possible benign features such as iso- or hyperenhancement were adjusted to a level lower; and thyroid nodules showing no enhancement were adjusted to C-TIRADS 3. Taking the pathological results as the gold standard, the receiver operating characteristic (ROC) curves of the C-TIRADS classification before and after the adjustment based on CEUS were plotted, and the diagnostic efficiency was compared. Results. The sensitivity, specificity, accuracy, and positive and negative predictive values of the C-TIRADS classification for the diagnosis of thyroid nodule malignancy before the adjustment based on the CEUS results were 83.6%, 63.8%, 74.4%, 72.7%, and 77.1%, respectively, and these values were 91.0%, 82.8%, 87.2%, 85.9%, and 88.9%, respectively, after the adjustment. The area under the ROC curve (AUC) was 0.737 and 0.869, respectively, showing a significant difference (Z = 3.288, P = 0.001 ). The diagnostic efficiency of C-TIRADS classification after the adjustment based on the CEUS results in both groups was improved compared with the result before the adjustment, and the difference in Group II was significant (Z = 2.931, P = 0.003 ). Conclusions. CEUS significantly improved the diagnostic performance in the adjustment of C-TIRADS 4 nodule classification, especially for the nodules which needs FNA recommended by the C-TIRADS.

The diagnostic value of contrast-enhanced ultrasound for cervical tuberculous lymphadenitis

OBJECTIVE: To investigate the value of contrast-enhanced ultrasound (CEUS) for the diagnosis of cervical tuberculous lymphadenitis (CTL). METHODS: The cohort study included 203 consecutive patients diagnosed with cervical lymph node. Before pathological or laboratory confirmation, all patients underwent CEUS examination, and the imaging findings were analyzed afterward. The diagnostic efficiency of the CEUS imaging findings of CTL was evaluated. RESULTS: Nighty-seven patients of the 203 (47.8%) were pathologically or laboratory confirmed with a CTL diagnosis while the remainder (52.2%) were diagnosed with non-tuberculous lymphadenitis. Regarding the imaging findings of CEUS, it was more common in CTL patients to find a pattern of heterogeneous enhancement inside the lymph nodes relative to non-tuberculous patients [81.44% (79/97) vs 15.09% (16/106), P < 0.01]. The sensitivity of the feature in diagnosis for CTL was 81.44% and the specificity was 84.91%, resepectively. Furthermore, a pattern of peripheral rim-like enhancement had been notable in CTL patients compared with non-tuberculous patients [86.60% (84/97) vs 12.26% (13/106), P < 0.01], associating with a diagnostic sensitivity of 86.60% and a specificity of 87.74% . When it came to the combination of both imaging findings mentioned above, the features were more prominent in CTL patients than compared with non-tuberculous patients [74.23% (72/97) vs 5.66% (6/106), P < 0.01], with a diagnostic sensitivity of 74.23% and a high specificity of 94.34% . Regarding area under curve (AUC) for the ROC analysis, the feature of internal heterogeneous enhancement, peripheral rim-like enhancement, and both features were 0.832, 0.872, and 0.843. CONCLUSIONS: CEUS patterns of heterogeneous enhancement and peripheral rim-like enhancement of lymph nodes are helpful characteristics for the diagnosis of CTL.

Bioinformatic Analysis Combined With Experimental Validation Reveals Novel Hub Genes and Pathways Associated With Focal Segmental Glomerulosclerosis

Background: Focal segmental glomerulosclerosis (FSGS) is a type of nephrotic syndrome leading to end-stage renal disease, and this study aimed to explore the hub genes and pathways associated with FSGS to identify potential diagnostic and therapeutic targets.Methods: We downloaded the microarray datasets GSE121233 and GSE129973 from the Gene Expression Omnibus (GEO) database. The datasets comprise 25 FSGS samples and 25 normal samples. The differential expression genes (DEGs) were identified using the R package “limma”. Gene Ontology (GO) function and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analyses were performed using the database for Annotation, Visualization and Integrated Discovery (DAVID) to identify the pathways and functional annotation of the DEGs. The protein–protein interaction (PPI) was constructed based on the Search Tool for the Retrieval of Interacting Genes (STRING) database and visualized using Cytoscape software. The hub genes of the DEGs were then evaluated using the cytoHubba plugin of Cytoscape. The expression of the hub genes was validated by quantitative real-time polymerase chain reaction (qRT-PCR) using the FSGS rat model, and receiver operating characteristic (ROC) curve analysis was performed to validate the accuracy of these hub genes.Results: A total of 45 DEGs including 18 upregulated and 27 downregulated DEGs, were identified in the two GSE datasets (GSE121233 and GSE129973). Among them, five hub genes with a high degree of connectivity were selected. From the PPI network, of the top five hub genes, FN1 was upregulated, while ALB, EGF, TTR, and KNG1 were downregulated. The qRT-PCR analysis of FSGS rats confirmed that the expression of FN1 was upregulated and that of EGF and TTR was downregulated. The ROC analysis indicated that FN1, EGF, and TTR showed considerable diagnostic efficiency for FSGS.Conclusion: Three novel FSGS-specific genes were identified through bioinformatic analysis combined with experimental validation, which may promote our understanding of the molecular underpinning of FSGS and provide potential therapeutic targets for the clinical management.

Diagnostic Efficacy of Ultrasound, Cytology, and BRAFV600E Mutation Analysis and Their Combined Use in Thyroid Nodule Screening for Papillary Thyroid Microcarcinoma

BackgroundUltrasound, cytology, and BRAFV600E mutation analysis were applied as valuable tools in the differential diagnosis of thyroid nodules. The aim of the present study was to evaluate the diagnostic efficiency of the three methods and their combined use in screening for papillary thyroid microcarcinoma (PTMC).MethodsA total of 1,081 patients with 1,157 thyroid nodules (0.5–1 cm in maximum diameter) classified as thyroid imaging reporting and data system (TIRADS) 4–5 were recruited. All patients underwent ultrasound, fine-needle aspiration (FNA) examination, and an additional BRAFV600E mutation test. TIRADS and Bethesda System for Reporting Thyroid Cytopathology (BSRTC) were adopted to judge the ultrasound and cytological results. The receiver operating characteristic (ROC) curve was established to assess the diagnostic values of different methods.ResultsOf the 1,157 nodules, 587 were benign and 570 were PTMCs. BRAFV600E mutation test had highest sensitivity (85.4%), specificity (97.1%), accuracy (91.4%), and area under the ROC curve (Az) value (0.913) among the three methods. The combination of BSRTC and BRAFV600E mutation analysis yielded a considerably high sensitivity (96.0%), accuracy (94.3%), and negative predictive value (95.9%) than either BSRTC or BRAFV600E mutation alone (P &lt; 0.0001 for all comparisons). Of all the methods, the combined use of the three methods produced the best diagnostic performance (Az = 0.967), which was significantly higher than that (Az = 0.943) for the combination of BSRTC and BRAFV600E mutation (P &lt; 0.0001). The diagnostic accuracy of the molecular method in the 121 nodules with indeterminate cytology was 90.1% (109/121), which was significantly higher than that of TIRADS classification, 74.4% (90/121) (P = 0.002).ConclusionThe combined use of ultrasound, cytology, and BRAFV600E mutation analysis is the most efficient and objective method for diagnosing PTMC. Both BRAFV600E mutation and TIRADS classification are potentially useful adjuncts to differentiate thyroid nodules, especially indeterminate samples classified as BSRTC III.

Hsa_circ_0005729 enhances the accuracy in diagnosing parathyroid carcinoma

Background: Parathyroid carcinoma (PC), often misdiagnosed as parathyroid adenoma (PA), is prone to local relapse due to the initial surgery being restricted to parathyroid lesions instead of en bloc resection of parathyroid lesions with negative incision margins. However, it is very challenging to distinguish PC from PA preoperatively; hence, this study investigated an effective biomarker for increasing accuracy in PC diagnosis. Method: First, differentially expressed the circular RNAs between three PC tissues and three PA tissues were screened by high-throughput circular RNA sequencing, and the expression of hsa_circ_0005729 was verified by qRT-PCR in 14 patients with PC and 40 patients with PA. Second, the receiver operating characteristic (ROC) curve and the area under the curve (AUC) were used to analyze the diagnostic efficiency of hsa_circ_0005729 in PC by combining with laboratory data. Third, RNF138 mRNA, the corresponding linear transcript of hsa_circ_0005729 was measured, and the relationship between hsa_circ_0005729 and RNF138 mRNA was analyzed in patients with PA and patients with PC. Results: Hsa_circ_0005729 expression was significantly higher in patients with PC than in patients with PA. Serum calcium (p = 0.045), alkaline phosphatase (ALP) (p = 0.048), and creatinine levels (p = 0.036) were significantly higher in patients with PC than in patients with PA. The AUC increased to 0.86 when hsa_circ_0005729 combined with serum calcium, creatinine, and ALP. In addition, hsa_circ_0005729 was positively correlated with RNF138 mRNA in patients with PA but not in patients with PC. Conclusion: The novel circular RNA hsa_circ_0005729 was found to have a higher expression in patients with PC, and indicating its usefulness for distinguishing PC from PA.