Journal of Clinical Laboratory Analysis最新文献

Background: Chalcones and heterocyclic compounds exhibit remarkably high activity in medicinal chemistry. In recent years, bis-chalcones have been reported to possess excellent anticancer activity. We synthesized a series of bis-heterocyclic chalcones via asymmetric chain synthesis, with the aim of making new discoveries in anticancer activity.

Methods: Bis-heterocyclic chalcones were synthesized via Claisen-Schmidt condensation and alkylation reactions. The inhibitory activities of the synthesized compounds against Huh-1, Huh-7, and HepG2 cell lines were evaluated using the CCK-8 assay. Furthermore, the mechanism of action of these compounds was explored through live/dead cell staining, flow cytometric analysis, and Western blotting experiments.

Results: Twelve bis-heterocyclic chalcone compounds were synthesized. All synthesized compounds were fully characterized by spectroscopic methods and evaluated for their cytotoxic activities against Huh-1, Huh-7, and HepG2 cell lines using the CCK-8 assay at concentrations ranging from 0 to 100 μM. Among them, derivative 3f exhibited the most potent cytotoxicity against Huh-7 (IC₅₀ = 8.40 μM) and Huh-1 (IC₅₀ = 6.75 μM), whereas HepG2 cells were most sensitive to compound 3d (IC₅₀ = 27.99 μM). The mechanisms underlying the antitumor effects of 3d and 3f were further investigated through live/dead cell staining, flow cytometry, and western blot analysis. The results demonstrate that both compounds effectively induce apoptosis in liver cancer cells.

Conclusions: Bis-heterocyclic chalcone compounds exhibit favorable inhibitory activity against hepatocellular carcinoma cells by inducing cell apoptosis, and thus can serve as a class of pharmaceutically active structural units for further in-depth research and screening.

Background: Discordant Vitamin B12 results can result from immunoassay interferences, potentially leading to unnecessary diagnostic procedures and misdiagnoses if not properly recognized.

Case presentation: A 76-year-old male with a history of IgA nephropathy, hypertension, and other comorbidities presented with unexpectedly elevated total Vitamin B12 levels despite no supplement use and normal hematological and biochemical parameters. Polyethylene glycol (PEG) precipitation demonstrated markedly reduced recovery, indicating the presence of macro-VitB12. Pretreatment with blocking reagents and exclusion of heterophilic antibodies confirmed the interference. The discrepancy observed between analytical platforms further supported this finding.

Conclusion: Vitamin B12 results should always be interpreted in conjunction with clinical findings. Polyethylene glycol (PEG) precipitation is a cost-effective and accessible method for detecting macro-VitB12, particularly in cases where analytical results are inconsistent with the clinical presentation.

Background: This study aimed to determine the potential interference of calcium dobesilate in two urinary total protein detection methods.

Methods: Urine samples from patients receiving calcium dobesilate were collected, and drug concentrations were quantified using liquid chromatography-tandem mass spectrometry. Paired-difference testing was used to assess if calcium dobesilate interfered with the total protein concentration measurement. The interference effect of calcium dobesilate on the detection method was evaluated using dose-effect experiments.

Results: In total, 22 24-h urine samples and 50 spot urine samples were collected. The calcium dobesilate concentrations ranged from 76 to 738 mg/L (median: 243 [161, 328] mg/L) and 60-2236 mg/L (median: 370 [195, 667] mg/L) in the 24-h and spot urine samples, respectively. Paired-difference testing showed that the total protein results deviated by 0.8%-12.1% and 13.0%-286.4% with the pyrogallol red-molybdate and benzethonium chloride methods, respectively, when the calcium dobesilate concentration was 4500 mg/L. The dose-response experiment with the benzethonium chloride method demonstrated a "J-shaped" effect; negative interference on the total protein concentration occurred at low and medium calcium dobesilate concentrations, but positive interference occurred at high calcium dobesilate concentrations.

Conclusions: Calcium dobesilate did not significantly interfere with the pyrogallol red-molybdate method but did significantly interfere with the benzethonium chloride method. The direction and degree of interference were related to the urinary protein and drug concentrations. Discontinuing calcium dobesilate for 1-2 days before obtaining a urine sample for total protein detection is recommended.

Background: Vascular smooth muscle cells (VSMCs) are crucial components of the arterial wall, playing a vital role in maintaining vascular integrity and function. Previous studies have identified HRD1 as a potential target for alleviating senescence in VSMCs. Ginsenoside Rb1 has been shown to counteract endothelial cell senescence triggered by H₂O₂ or oxidized LDL.

Methods: In this study, Rb1 was investigated to determine if it could protect VSMCs from cholesterol-induced senescence. VSMCs were pretreated with Rb1 and subsequently exposed to cholesterol to evaluate its effects on SA-β-gal activity, reactive oxygen species (ROS) generation, cell viability, and STING pathway activation.

Results: Rb1 treatment significantly reduced the proportion of SA-β-gal-positive cells induced by cholesterol. Moreover, Rb1 suppressed the activation of endoplasmic reticulum (ER) stress markers and inhibited STING signaling. HRD1 knockdown abrogated the Rb1-mediated reduction of ROS production. Similarly, both Rb1 and an STING inhibitor decreased cholesterol-induced mitochondrial ROS (MitoSOX) levels.

Conclusion: These findings indicate that Rb1 protects VSMCs against cholesterol-induced senescence by preserving HRD1 expression, mitigating ER stress, and maintaining mitochondrial function. Therefore, Rb1 holds therapeutic potential for preventing vascular diseases associated with VSMC senescence by modulating the HRD1 and STING pathways.

Background: Soluble immune checkpoints (sICs) are emerging as possible serum and plasma biomarkers in cancer and immune-mediated diseases, but little is known about the impact of the matrix type in sIC detection. This study aimed to assess whether sIC measurements are comparable between serum and EDTA-plasma samples.

Methods: A cohort of 38 healthy subjects was enrolled. A multiplex bead-based assay was used to evaluate a panel of 17 sICs (CD137, 4-1BBL, CD27, CTLA4/CD152, CD80, CD40, CD40L, GITR, GITRL, ICOSL, IDO, LAG3, PD-1, PD-L1, PD-L2, TIM3, and VISTA) in paired serum and plasma-EDTA samples. The detection frequencies, concentrations, and correlations of each sIC were analyzed by comparing the two matrices.

Results: Soluble CD137, CD152, CD40, and LAG3 were detected more frequently in plasma, while soluble CD40L was detected predominantly in serum. The concentrations of soluble 4-1BBL, CD27, PD-1, VISTA were higher in plasma, while the concentrations of soluble PD-L2 were higher in serum. The concentrations of soluble CD80, GITR, GITRL, ICOSL, IDO, and TIM3 were comparable between serum and plasma. Soluble CD27, CD80, GITRL showed a significant positive, slight correlation between plasmatic and serum concentrations.

Conclusion: Except for soluble CD80, the detection of the other sICs by the bead-based assay was influenced by the matrix type. The evaluation of the best matrix for sICs should be considered before starting clinical studies.

Objective: Cryptosporidium spp. is an important pathogen responsible for severe diarrheal illness, especially in children, and is transmitted by various modes. The present work is aimed at categorizing Cryptosporidium spp. infection and determining associated risk factors by ML on a known dataset of diarrhea among children.

Materials and methods: For classification, we used random forest and bagging CART trees. Model discrimination was measured by accuracy, balanced accuracy, sensitivity, specificity, positive predictive value, negative predictive value, and F1-score. Then, a 5-fold cross-validation method was used to verify the reliability of the model. Importance values were also calculated to identify the most important risk factors for infection.

Results: The bagged CART model emerged as the best among the models applied, with slightly better classification. For this model, performance metrics were: accuracy (87.2%), balanced accuracy (56.3%), sensitivity (97.2%), specificity (15.4%), positive predictive value (89.3%), negative predictive value (42.9%), F1-score (93.0%). As shown by the variable importance analysis, the strongest risk factor was the number of people in the household (people ≥ 5), which represented a higher risk of infection in crowded housings. Sources of water also came up as an important environmental factor; plain tap water and pipe-line water appeared to be major causes of transmission.

Conclusion: Such results indicate that waterborne transmission is the main route of Cryptosporidium spp.

Infection: These findings underscore the importance of water quality improvements, including efforts to address water disinfection, particularly in areas with household crowding and inadequate sanitation access.

Background: Prostate-Specific Antigen (PSA) is widely used for Prostate Cancer (PCa) screening, but its low specificity often leads to false-positive results and unnecessary biopsies. To address this issue, we introduced Age-Adjusted Prostate-Specific Antigen Density (A-PSAD) as a potential biomarker to improve screening accuracy for PCa and Clinically Significant Prostate Cancer (csPCa).

Methods: In this study, 663 patients who underwent prostate biopsy between 2020 and 2024 were included. The diagnostic performance of A-PSAD, PSAD, and other biomarkers was compared. Key variables, including age, diabetes history, FPSA, prostate volume, and A-PSAD, were selected using Lasso regression to develop a nomogram prediction model (Nomo1). Additionally, the PI-RADS score was incorporated into a second model (Nomo2).

Results: A-PSAD outperformed PSAD in PCa and csPCa screening. For PCa, A-PSAD had a higher AUC (0.753 vs. 0.732) with 71.86% sensitivity and 70.69% specificity, compared to PSAD's 74.87% and 63.15%. For csPCa, A-PSAD's AUC was 0.731 versus 0.708, with sensitivities of 76.27% and 57.61%. The nomogram model, based on Lasso-selected variables, achieved AUCs of 0.796 and 0.812 in the training and validation sets, with C-indices of 0.7870 and 0.7935, indicating good predictive performance. Decision curve analysis showed high clinical benefit across most risk thresholds. Incorporating PI-RADS, the Nomo2 model improved diagnostic performance in high-risk patients (AUC 0.8202) without significant difference from Nomo1 (AUC 0.8198, p > 0.05).

Conclusion: A-PSAD offers a better balance between sensitivity and specificity compared to PSAD, reducing false positives and unnecessary biopsies, and presents a promising tool for personalized PCa screening.

Background: The scale-up of molecular assays for diagnosing emerging pathogens has increased in low-and-middle-income countries (LMICs) since the advent of COVID-19. We herein evaluated the diagnostic concordance of three different assays for SARS-CoV-2 in Cameroon.

Methods: A laboratory-based comparative study was performed on nasopharyngeal samples collected between March-2020 to March-2023 from the biobank of Chantal Biya International Reference Centre (CIRCB), Yaoundé-Cameroon. Samples were analyzed using DaAn Gene (N/ORF1ab-genes), ThermoFisher (N/ORF1ab/S-genes), and GeneXpert (N2/E-genes). Validated cycle thresholds (CT) for positivity were CT < 37 for DaAn Gene/ThermoFisher and CT < 40 for GeneXpert. Cohen's Kappa coefficient evaluated diagnostic concordance with DaAn Gene as reference.

Results: We analysed 249 samples (55.8% males, median-age [IQR], 36 [27-50] years including 21.3% symptomatic participants). Overall positivity rates (median [IQR]) were 55.0% (CT: 30.6 [23.1-35.5]); 53.4% (CT: 26.6 [21.2-30.9]); 22.1% (CT: 32.7 [26.9-36.1]) for GeneXpert, DaAn Gene and ThermoFisher respectively. GeneXpert showed stronger concordance with DaAn Gene (83.1%; k = 0.66, 95% CI: 0.57-0.75) than ThermoFisher (67.9%; k = 0.38, 95% CI: 0.29-0.47). At validated thresholds, GeneXpert showed higher positive agreement with DaAn Gene (85.0%, 113/133) as compared to ThermoFisher (41.3%, 55/133), while maintaining comparable negative agreement (81.0% [GeneXpert] and 98.3% [ThermoFisher]). At low CTs (< 20) however, positive agreement with DaAn Gene was high for GeneXpert (100%, 15/15) and ThermoFisher (93.3%, 14/15).

Conclusion: GeneXpert exhibits superiority over ThermoFisher in detecting cases of COVID-19. As expected, agreement between two- and three-genes assays at CT < 20 was excellent, suggesting interoperability of these platforms during outbreaks for high viral loads cases. However, two-genes assays may be decisive to guide decision-making for effective public health response while facing intermediate to low-level viral loads in LMICs.

Introduction: In South Korea, low-resolution chromosomal microarray analysis (LR-CMA) is frequently used as a screening tool to identify chromosomal anomalies in asymptomatic newborns. However, its clinical utility remains controversial.

Methods: We retrospectively analyzed 99 asymptomatic newborns who underwent diagnostic CMA following abnormal LR-CMA screening results at a single tertiary hospital between 2019 and 2024. Clinical features, copy number variant (CNV) findings, and follow-up outcomes were assessed.

Results: Among the 99 patients (57.6% male), the median ages at the initial and last visits were 0.3 and 1.0 years, respectively. A total of 171 CNVs were identified, of which 85 (49.7%) were classified as pathogenic or likely pathogenic. Overall, 70 of 99 (70.7%) patients harbored microduplication or deletions with syndromic implications. Developmental delay was identified in 10 (10.1%) patients. Notably, no significant associations were found between CNV pathogenicity and prenatal history, presence of anomalies, developmental delays, or growth parameters.

Conclusions: Although a high rate of clinically significant CNVs was detected through LR-CMA screening, only a minority of asymptomatic newborns exhibited developmental concerns within the observed follow-up period. These findings suggest that while LR-CMA may identify genetic alterations of interest, its routine use in asymptomatic newborns warrants careful consideration of clinical relevance and potential psychosocial impact.