Annals of clinical and laboratory science最新文献

Objective: The pathogenesis of intrauterine adhesions remains unclear, with angiotensin potentially contributing to their progression.

Methods: A rat model of intrauterine adhesion (IUA) was constructed. Histomorphological analysis of endometrial architecture was performed using hematoxylin-eosin (HE) staining. Protein expression profiles of NLRP3, IL-1β, α-smooth muscle actin (α-SMA), and E-cadherin were quantified through Western blot analysis. PCR detection was performed using primer sequences specific to the factors of interest.

Results: The rat IUA model exhibited characteristic uterine wall adhesions, marked inflammatory infiltration, and significantly reduced vimentin expression compared to sham-operated controls. Systemic analyses revealed Ang II's concentration-dependent promotion of IUA progression, with 10^-5-10^-7 mol/L doses significantly elevating endometrial epithelial cell (EEC) proliferation (p<0.05), collagen I/III secretion (p<0.05), and EMT marker dysregulation. Mechanistically, Ang II activated the NLRP3 inflammasome pathway, driving IL-1β overexpression and pyroptosis, with maximal adhesion severity at 10^-5 mol/L. NLRP3 agonism exacerbated inflammatory responses, while siRNA-mediated NLRP3 knockdown restored E-cadherin expression and attenuated N-cadherin, effectively reversing EMT progression.

Conclusion: This study demonstrates that angiotensin II drives intrauterine adhesion progression through NLRP3 inflammasome-mediated inflammatory escalation and fibrotic remodeling. The identified Ang II-NLRP3 axis may offer a dual therapeutic target for mitigating both pathological inflammation and endometrial fibrosis in adhesion prevention.

Objective: This study aims to compare the diagnostic performance of colloidal gold immunochromatographic assay (GICA) with culture-based screening for antepartum detection of Group B Streptococcus (GBS).

Methods: This prospective study was conducted in a tertiary hospital in South China, performing GBS screening on women at 35-37 weeks of gestation. GICA and direct bacterial culture were separately performed on paired rectovaginal swabs. Decisions about intrapartum antibiotic prophylaxis (IAP) were based on the GBS positive results by either GICA or culture.

Results: The detection rate for GBS colonization was 7.5% (31/414) by the culture and 16.4% (68/414) by the GICA, with significant difference (p<0.001). Kappa value between the two methods was 0.516 (p<0.001), suggesting moderate concordance. Against the reference standard of direct culture, the GICA showed a sensitivity of 90.3%, specificity of 89.6%, positive predictive value (PPV) of 41.2%, negative predictive value (NPV) of 99.1%, and accuracy of 89.6%. Among individuals with positive GICA, the culture-positive group was associated with significantly higher proportion of IAP and lower relative risk of adverse pregnancy outcomes compared to the negative group.

Conclusion: The GICA demonstrates 90% sensitivity and specificity in detecting maternal GBS colonization compared to culture. This rapid test may be considered as a promising alternative, particularly for emergency labor and large-scale screening in resource-limited settings. However, the clinical overuse of IAP should be concerned.

Objective: Isolated Myeloid sarcoma (IMS) of breast is extremely rare and current treatment approaches for Acute Myeloid Leukemia (AML) secondary to breast IMS are even rarer. It is worthwhile to explore the clinical features of this disease and the promising treatment strategy.

Case report: We described one case of IMS that occurred in the breast in a 37-year-old female, who progressed to RUNX1::RUNX1T1 positive AML in less than two years. Then, she was treated with Venetoclax plus hypomethylating agents, which showed favorable response.

Discussion: She has remained in complete remission (CR) to date after the Venetoclax-based treatment.

Conclusions: Breast IMS is rare but highly progressive to AML. Imaging and histopathology are key tools in diagnosing it and Venetoclax-based regimens may be a promising treatment strategy.

Objective: The mechanism of auraptene (AUR) in the treatment of acute myocardial infarction (AMI) was explored through in vitro and in vivo experiments combined with network pharmacology technology.

Methods: Network pharmacology and molecular docking were used to predict the potential targets and related pathways of AUR in AMI. AMI was induced by ligating the left anterior descending coronary artery of Sprague-Dawley rats. Prior to modeling, AUR (50 mg/kg) was administered continuously for one week. AMI in rats was assessed by ultrasonic electrocardiogram, TTC staining, serum myocardial enzyme, hematoxylin-eosin staining, TUNEL staining, and apoptotic protein detection. Endoplasmic reticulum stress (ERS) in rat myocardium was evaluated by dihydroethidium staining and measurement of ERS-related proteins. An AMI cell model was established in oxygen and glucose deprivation (OGD)-induced H9C2 rat cardiomyocytes. Interventions with AUR, ERS agonist tunicamycin (TM), or epithelial growth factor (EGF) (EGFR agonist) were applied to H9C2 cells induced by OGD. Cell damage was evaluated using CCK-8 assay, lactic dehydrogenase measurement, and apoptotic protein detection. ERS in H9C2 cells was evaluated using the ER-Tracker Red molecular probe and ERS-labeled proteins. The expression of EGFR/extracellular regulated protein kinases (ERK) signaling pathway-related proteins was detected by western blot.

Results: Network pharmacology and molecular docking suggest that EGFR may be a potential target for AUR in AMI, with the ERK signaling pathway identified as a crucial pathway. In vivo, AUR preconditioning significantly improved myocardial injury in AMI rats and inhibited ERS and EGFR/ERK signaling pathway activities in myocardial tissue. In vitro, AUR pretreatment reduced ERS induced by OGD in H9C2 cells. Compared to the OGD+AUR group, the OGD+AUR+TM group showed significantly increased cell damage and ERS level (P<0.05). Compared with the OGD+AUR group, the activity of the EGFR/ERK signaling pathway, ERS level, and the degree of cell damage in the OGD+AUR+EGF group were significantly improved (P<0.01).

Conclusion: AUR inhibits ERS by regulating the EGFR/ERK signaling pathway, thus improving outcomes in AMI rats.

Objective: Liquid chromatography combined with tandem mass spectrometry (LC-MS/MS) is the gold standard for the analysis of steroids including 11-deoxycortisol, androstenedione, testosterone, 17-hydroxyprogesterone, and dehydroepiandrosterone (DHEA) in serum or plasma. In our hospital we observed an occasional problem in analysis of testosterone in some patients when blood was collected in Beckton-Dickinson serum separator gel tubes (BD gel tubes). Therefore, we investigated this issue.

Methods: Blood was collected from 10 volunteers (five males and five females) simultaneously in plain red top tubes (no gel), Greiner serum separator gel tubes (Greiner gel tube), and BD (Beckton-Dickinson) gel tubes followed by extraction of steroids from serum, reconstitution with mobile phase, and subsequent analysis of various steroids using LC-MS/MS in a single run.

Results: No interfering peak was observed when blood was collected in plain red top tube for all five steroids. However, we observed clinically significant interference with low testosterone values when blood specimens were collected in BD gel tubes. For DHEA, we observed interference when blood was collected in BD gel tubes and Griner tubes. Other steroids were not affected.

Conclusions: We concluded that BD serum separator gel tube may not be suitable for analysis of testosterone in women and DHEA for both men and women.

Objective: The stricter Clinical Laboratory Improvement Amendments (CLIA) 2024 guidelines introduced narrower Total Allowable Error (TEa) limits, posing challenges for clinical laboratories in maintaining analytical quality. This study evaluated the effectiveness of Six Sigma metrics in assessing the performance of routine biochemical tests using these updated criteria.

Methods: This retrospective study analyzed the internal quality control (IQC) data for 22 biochemical analytes across four instruments in a high-throughput laboratory. Performance was assessed using Sigma metrics calculated based on the CLIA 88 and CLIA 2024 criteria, with classifications into poor (<3 Sigma), acceptable (3-6 Sigma), and excellent (>6 Sigma) categories. İnstrument-specific variability and control levels (normal and pathological) were also analyzed.

Results: Under the CLIA 2024 guidelines, only 22.16% of the analytes achieved excellent performance (>6 Sigma), compared to 49.43% under CLIA 88. No significant differences were observed between instruments, indicating consistent analytical performance across platforms. Normal control levels (Control Level 1) exhibited greater variability (median Sigma: 4.76, range: 1.19-13.34) compared to pathological controls (Control Level 2) (median Sigma: 4.72, range: 1.22-10.22), reinforcing the impact of control level differences on analytical precision. CRP, CK, and Bilirubin were the highest-performing tests, consistently maintaining high Sigma values above the acceptable threshold. In contrast, Albumin, Urea, and GGT exhibited the lowest Sigma performance.

Conclusions: The transition to stricter CLIA 2024 guidelines significantly affects the analytical performance of biochemical tests, highlighting vulnerabilities in routine laboratory operations. Adopting advanced automation, tailored QC protocols, and modern analytical tools is essential to enhance diagnostic precision and ensure compliance with evolving regulatory demands.

Objective: MicroRNAs (miRNAs) have been recognized as important candidates for diagnostic, prognostic, and predictive biomarkers for many human cancer types, including gastric cancer. Previous studies have found that the expression level of miR-214 was increased in gastric cancer tissues, and it was able to promote the proliferation, migration, and invasion of gastric cancer cells. In the present study, we aimed to investigate its clinical significance and prognostic value in gastric cancer.

Methods: A total of 133 pairs of gastric cancer tissues and corresponding adjacent normal gastric tissues were obtained from 133 patients diagnosed with gastric cancer. Quantitative real-time PCR (qRT-PCR) assays were carried out to detect the expression level of miR-214. Associations between tissue miR-214 expression level and clinicopathological parameters were evaluated using Chi square test. Overall survival was calculated, and survival curves were plotted using the Kaplan-Meier method; differences between groups were compared using log-rank tests. Multivariate analysis using the Cox proportional hazards regression model was performed to assess the prognostic value of miR-214 expression.

Results: We found that expression level of miR-214 was significantly higher in the gastric cancer tissues than in the adjacent normal gastric tissues (P<0.001). The expression levels were significantly associated with depth of invasion (P=0.022), differentiation degree (P=0.009), lymph node metastasis (P<0.001), and TNM stage (P<0.001). The patients with high miR-214 expression level showed reduced overall five-year survival compared to those with low miR-214 expression (log-rank test P=0.015). Furthermore, multivariate regression analysis showed that tissue miR-214 expression level was an independent prognostic factor overall survival (HR=2.102, 95% CI: 1.572-8.822, P=0.033) in gastric cancer.

Conclusions: Our results indicated that miR-214 was closely related to the occurrence, progression, and metastasis of gastric cancer, and overexpression of miR-214 in tissues might serve as a potential prognostic biomarker for gastric cancer.

Objective: Atherosclerosis (AS) contributes significantly to the development of cardiovascular disease. Despite recent advances in medical treatment, the outcomes for patients with AS are still unsatisfactory. This study aims to enhance our understanding of AS and pave the way for the development of more effective therapeutic approaches to improve patient outcomes and ultimately reduce the burden of this devastating condition.

Methods: An AS cell model was established using oxidized low-density lipoprotein (OX-LDL) administration to human vascular smooth muscle cells (HVSMCs), followed by injection with oe-NC, oe-FUS, oe-Sfrp5, or oe-FUS+ oe-Sfrp5. Western blot, Transwell assay, CCK8 assay, Oil Red O staining assay, and ELISA were used to elucidate the mechanisms of the FUS/Sfrp5/Wnt5 pathway in our model of AS.

Results: In this study, both FUS and Sfrp5 inhibited Wnt5a expression. In addition, FUS inhibited lipid droplet formation and migration capacity of OX-LDL-induced HVSMCs, and FUS and Sfrp5 may have a synergistic effect in controlling Wnt5a in AS. These results highlight the potential therapeutic value of targeting the FUS and Sfrp5 pathway to control AS progression.

Conclusion: The findings suggest that FUS and Sfrp5 may have a synergistic effect in controlling Wnt5a in AS.