Clinica Chimica Acta最新文献

Pancreatic cancer is a highly fatal malignancy due to poor early detection rate and resistance to conventional therapies. This review examines the potential for liquid biopsy as a transformative technology to identify diagnostic and therapeutic targets in pancreatic cancer. Specifically, we explore emerging biomarkers such as exosomal non-coding RNAs (ncRNAs), circulating tumor DNA (ctDNA), and circulating tumor cells (CTCs). Tumor-derived exosomes contain nucleic acid and protein that reflect the unique molecular landscape of the malignancy and can serve as an alternative diagnostic approach vs traditional biomarkers like CA19-9. Herein we highlight exosomal miRNAs, lncRNAs, and other ncRNAs alongside ctDNA and CTC-based strategies, evaluating their combined ability to improve early detection, disease monitoring and treatment response. Furthermore, the therapeutic implications of ncRNAs such as lncRNA UCA1 and miR-3960 in chemoresistance and progression are also discussed via suppression of EZH2 and PTEN/AKT pathways. Emerging therapeutic strategies that target the immune response, epithelial-mesenchymal transition (EMT) and drug resistance are explored. This review demonstrates a paradigm shift in pancreatic cancer management toward personalized, less invasive and more effective approaches.

Copy number variations (CNVs) in the 7q11.2 and 22q11.2 chromosomal regions are major contributors to genetic disorders such as Williams-Beuren syndrome and 22q11.2 deletion/duplication syndromes. These disorders are characterized by facial anomalies, growth retardation, intellectual disabilities, and lethal cardiovascular abnormalities. Despite the development and clinical application of various rapid molecular tests, each has significant limitations. We developed a multiplex chip-based droplet digital PCR (ddPCR) method for detecting microdeletions and microduplications in the 7q11.2 and 22q11.2 regions. We evaluated its linearity and reproducibility and tested it on 100 clinical patients with congenital heart defects to further verify its clinical applicability. We successfully developed a method capable of simultaneously detecting four types of CNVs in a single assay, demonstrating high accuracy and reproducibility. Compared to traditional methods, our approach depicted 100% concordance for positive and negative results. Additionally, our method accurately quantified target gene concentrations, allowing for precise evaluation of CNVs in the target regions. This study introduces a rapid, technically straightforward, and efficient quantitative chip-based ddPCR method for the detailed classification of CNVs in the 7q11.2 and 22q11.2 regions. Our findings indicated that chip-based ddPCR can be seamlessly implemented as a first-line screening tool in routine diagnostics.

Objectives: This study investigates the optimal number of quality control (QC) events per day in two scenarios: high-sensitive troponin, requiring three levels of QC materials, and creatinine, with two levels. The aim is to explore how different QC rules and sigma metric values affect the frequency of QC events, considering both analytical performance and the clinical impact of potential measurement errors as severity of harm.

Methods: Risk-based QC calculations were performed using the QC Constellation tool. Four QC rule schemes (1-3 s, 1-3 s/2-2 s, 1-3 s/2-2 s/R-4 s, 1-3 s/2-2 s/R-4 s/4-1 s) were tested for high-sensitive troponin (catastrophic harm) and creatinine (serious harm). Sigma metric values from 3 to 6 were evaluated to determine maximum run sizes. QC event frequency was estimated for a hypothetical laboratory processing 1,000 samples daily.

Results: Maximum run sizes decreased as sigma metric values declined. Correspondingly, the number of QC events per day increased as sigma metric values decreased. For high-sensitive troponin, with its catastrophic severity of harm related to potantial error, more frequent QC was necessary compared to creatinine.

Conclusions: Analytical performance and severity of harm significantly influence the required frequency of QC events. Laboratories must consider both factors when designing QC strategies to balance patient safety with operational efficiency. For analytes with high severity of harm, achieving higher sigma metrics is critical to maintain feasible and cost-effective QC practices. A hybrid QC approach combining risk-based and patient-based methods may optimize QC strategies, but standardization of sigma metric calculations is still needed.

With the frequent outbreaks of viral diseases globally, accurate and rapid diagnosis of viral infections is of significant importance for disease prevention and control. The CRISPR-Cas combined biosensing strategy, as an emergent nucleic acid detection technology, exhibits notable advantages including high specificity, elevated sensitivity, operational simplicity, and cost-effectiveness, thereby demonstrating significant potential in the domain of rapid viral diagnostics. This paper summarizes the principles of the CRISPR-Cas system, the novel biotechnologies, and the latest research progress in virus detection using the combined biosensing strategy. Additionally, this paper discusses the challenges faced by CRISPR-Cas biosensing strategies and outlines future development directions, which provides a reference for further research and clinical applications in the rapid diagnosis of viral infections.

Objective: Pancreatic cancer (PC) is a highly aggressive malignancy with poor prognosis and high mortality rate. Identifying reliable biomarkers for the early diagnosis and treatment is urgently needed. This study aims to comprehensively evaluate the diagnostic and prognostic value of DUPAN-2 in PC through a meta-analysis.

Methods: We systematically searched PubMed, Embase, and other databases for studies related to DUPAN-2 and its prognostic and diagnostic relevance in PC, covering publications up to August 2024. We used pooled hazard ratios (HRs) to evaluate the prognostic value of DUPAN-2 in PC, the summary receiver operating characteristic (SROC) curve and the area under the curve (AUC) to assess diagnostic performance, while pooled odds ratios (ORs) analyzed associations with clinicopathological features.

Results: A total of 22 studies involving 4765 patients were included in this meta-analysis, with 11 studies focusing on diagnostic analysis, 10 on prognostic analysis, and 3 on clinicopathological features. The diagnostic meta-analysis revealed a pooled sensitivity of 0.63 (95 % CI: 0.56-0.69), a pooled specificity of 0.98 (95 % CI: 0.95-0.99), and an AUC of 0.83 (95 % CI: 0.79-0.86). Subgroup analysis indicated that a DUPAN-2 threshold at 150 U/mL achieved the highest diagnostic performance. The prognostic meta-analysis demonstrated that elevated DUPAN-2 levels were associated with poorer OS (HR = 1.70, 95 % CI: 1.36-2.14) and PFS (HR = 1.33, 95 % CI: 1.14-1.56). Additionally, the clinicopathological features meta-analysis showed that elevated DUPAN-2 levels were associated with vascular invasion (OR = 3.48, 95 % CI: 1.26-9.59), while normalized DUPAN-2 levels were associated with higher resectability (OR = 0.57, 95 % CI: 0.36-0.90) and lower N-stage (OR = 0.39, 95 % CI: 0.24-0.63) CONCLUSION: Serum DUPAN-2 demonstrates significant potential as a biomarker for diagnosis and prognosis in patients with PC.

Objective: Laboratory extensively applied enzyme-linked immunosorbent assay (ELISA) to measure anti-phospholipase A2 receptor antibodies (PLA2R-abs) since its diagnostic significance on PLA2R related primary membranous nephropathy (PLA2R-related pMN) was discovered. However, PLA2R-abs determined by ELISA (PLA2R-ELISA) could infrequently yield inconclusive results, specifically a grey-zone defined as PLA2R-abs ranging from 2 to 20 RU/mL. Recently, researchers suggested that double-check grey-zone PLA2R-abs by indirect immunofluorescence (IIF) could improve diagnostic accuracy. We evaluated the diagnostic performance of PLA2R-IIF in assessing PLA2R-related pMN and summarized clinicopathological characteristics of grey-zone population to provide more evidence for clinical practice.

Methods: Data on demographics, serology and pathology of patients with PLA2R-ELISA grey-zone results and a native kidney biopsy at Peking Union Medical College Hospital from September 2020 to April 2023 were reviewed. Grey-zone samples were analyzed using PLA2R-IIF. Negative results were defined as no fluorescence and positive results were graded according to fluorescence intensity.

Results: This study included a total of 52 grey-zone patients divided into pMN group (n = 36, 69 %) and non-pMN group (n = 16, 31 %) according to renal pathology reports. The pMN patients had higher PLA2R-abs and lower serum creatinine compared to the non-pMN patients (P = 0.003, P < 0.001). No statistically significant differences were observed in 24-hour urine protein and albumin between the two groups. Multiple pathological types were identified in the non-pMN group. The sensitivity and specificity of PLA2R-IIF in PLA2R-ELISA grey-zone population were 72 % and 88 %, respectively, with a total consistent rate of 77 % and a positive predictive value of 93 %.

Conclusion: Both pMN and non-pMN patients presented grey-zone PLA2R-ELISA results. It was necessary to perform PLA2R-IIF to assist in the diagnosis of patients with PLA2R-ELISA grey-zone results.

Background: Corticosteroid-binding globulin (CBG) modulates tissue cortisol availability via modification of cortisol:CBG binding affinity in response to multiple factors, including neutrophil elastase (NE) cleavage of the reactive centre loop (RCL), converting high affinity CBG (haCBG) to low affinity CBG (laCBG). In vitro, glycosylation of the RCL at Asn347 affects NE cleavage susceptibility. To date, no direct measurement of laCBG, which would verify NE cleavage, has been reported.

Objective: To measure serum laCBG in septic shock patients by mass spectrometry to confirm NE cleavage in vivo, with Asn347 site glycosylation profiling to determine its impact on NE cleavage, and determine effect of %laCBG on septic shock clinical outcome.

Methods: Serum from septic shock patients from a tertiary hospital intensive care unit was analysed by mass spectrometry for CBG affinity forms and Asn347 glycosylation profile, following CBG immunoprecipitation. Data was correlated with septic shock clinical outcome.

Results: N-terminal peptide of NE cleaved RCL (AVLQLNEEGVDTAGSTGV) was consistently detected in patient serum, although at low concentrations. Mean %laCBG/total CBG was 0.23 % in septic shock (range = 0.07-0.74 %, SD = 0.12 %); in comparison healthy controls mean %laCBG was 0.04 % (range 0.02-0.08 %, SD = 0.03 %). There was a negative correlation between %laCBG and serum concentrations of CBG with triantennary Asn347 glycans; TS3 (r = -0.190, p = 0.040) and TS3F (r = -0.252, p = 0.006).

Conclusions: NE cleaved CBG (laCBG) is present in septic shock patient serum, and %laCBG correlates with Asn347 glycosylation occupancy and composition. However, serum %laCBG did not correlate with septic shock clinical outcome.

Cervical cancer (CC) is the fourth most common cancer among women worldwide, following breast, colorectal, and lung cancers. Each year, it accounts for approximately 600,000 new cases and 340,000 deaths. Early-stage cervical cancer is treatable with surgery and chemoradiotherapy (CCRT). However, treatment for metastatic cervical cancer is limited, with bevacizumab combined with chemotherapy being one of the few options, though survival rates remain low. Currently, the diagnosis of cervical cancer primarily relies on Pap smears and colposcopy. Although these methods are essential for detection, they are costly, labor-intensive, and require significant resources. Therefore, there is an urgent need to identify effective biomarkers that can detect cervical cancer at an early stage, improving both the accuracy of diagnosis and the efficacy of treatment. Although numerous cervical cancer biomarkers have been identified for the cervical cancer thanks to advances in technology. In recent times, electrochemical methods have proven to be particularly effective in cervical cancer detection. In this paper, we reviewed the important cervical cancer biomarkers and their detection through electrochemical biosensors, which offer advantages such as higher sensitivity, affordability, and ease of analysis. Furthermore, we discussed the limitations and future prospects of electrochemical biosensors in this field.

Background and aims: Cerebral small vessel diseases (CSVDs) are a set of conditions that affect the small blood vessels in the brain and can cause severe neurological pathologies such as stroke and vascular dementia. The most common monogenic CSVD is cerebral autosomal dominant arteriopathy with subcortical infarcts and leukoencephalopathy (CADASIL) which is caused by mutations in NOTCH3. However, only 15-20% of CADASIL cases referred for genetic testing have pathogenic mutations in NOTCH3. We hypothesise that other monogenic causes of CSVD may be causing a CADASIL-like CSVD phenotype.

Methods: To test this, we performed whole exome sequencing for 50 individuals suspected of having CADASIL, but did not exhibit a disease-causing mutation in NOTCH3, and applied targeted analysis of all monogenic forms of CSVD.

Results: This analysis identified three mutations affecting the Collagen type IV genes in three individuals likely to be causative of CSVD.

Conclusions: This suggests that screening for all monogenic forms of CSVD when one monogenic form is clinically suspected may improve diagnosis in clinically suspected monogenic CSVD. However, despite these findings, the majority of NOTCH3 negative CSVD cases did not have candidate mutations in known CSVD genes, suggesting that additional genetic factors contributing to the disease are yet to be identified.