Annals of clinical and laboratory science最新文献

Objective: Chronic diabetic nephropathy (CDN) is one of the common complications of the chronic diabetes. The FGF23/FGFR3-mediated signaling pathway is involved in CDN. Whether miR-190a-3P participates in CDN through regulation of FGFR3 remains to be elucidated. The present study evaluated miR-190a-3P's effect on CDN.

Methods: Mice were divided into a control group (NC group), a CDN group, and CDN+miR-190a-3P antagonist group, followed by analysis of miR-190a-3P and FGFR3 level by qRT-PCR, FGFR3, and Tubulin protein level by Western blot as well as blood glucose, serum creatinine (Cr), and urea nitrogen (BUN) using an automatic biochemical analyzer.

Results: Compared with the NC group, the CDN group had a significantly higher miR-190a-3P level in kidney tissue, while the FGFR3 mRNA level was lower (P<0.01). FGFR3 was a target gene of miR-190a-3P. miR-190a-3P levels in the CDN+miR-190a-3P antagonist group were significantly reduced compared with the CDN group (P<0.05). Meanwhile, FGFR3 protein levels in miR-190a-3P antagonist group were significantly increased (P<0.05). Compared with the NC group, the CDN group showed significantly reduced blood glucose level and elevated BUN and Cr level (P<0.01). However, CDN+miR-190a-3P antagonist group showed significantly increased blood glucose and reduced BUN and Cr level compared with CDN group (P<0.05).

Conclusion: miR-190a-3P can directly bind to the 3'-UTR of FGFR3 mRNA and reduce FGFR3 protein levels, contributing to the occurrence of CDN.

Objective: To investigate the protective effect of Jiawei Shuxin Tang (JWSXT) on myocardial ischemia-reperfusion injury (MIRI) rats with Qi deficiency and blood stasis and its correlation with neuregulin-1 (NRG-1).

Methods: 30 rats were used to establish the model of Qi deficiency and blood stasis, and were randomly divided into control group, MIRI model group, and JWSXT group. The traditional Chinese medicine (TCM) phenotypes, echocardiographic performance, myocardial reperfusion area, myocardial infarction area, HE staining, and NRG-1 content changes of rats in each group were compared one week after treatment.

Results: We found that compared with the model group, rats in the JWSXT group showed increased activity as well as improved tongue texture and lip color (P<0.05); LVEF and LVFS increased in the JWSXT group (P<0.05; P<0.05). The percentage of infarct area and the percentage of myocardial ischemia reperfusion area in the JWSXT group was significantly reduced (P<0.05; P<0.05). Compared with the control group, the remaining NRG-1 content in myocardial tissues of rats in the two groups was decreased (P<0.01), and NRG-1 content in the JWSXT group was increased (P<0.05) compared with the model group.

Conclusions: JWSXT can improve reperfusion injury in rats with Qi deficiency and blood stasis infarction, which correlates with NRG-1.

Objective: The consistency of plasma catecholamines and metanephrines measurements is crucial for the screening and diagnosis in pheochromocytomas and paragangliomas (PPGLs). There are many commercially available kits for plasma catecholamines and metanephrines testing, but their performance and consistency information are unknown. This study evaluated the performance of three mainstream commercially available LC-MS/MS kit methods and evaluated their consistency.

Methods: Performance validation of three kit methods was performed according to Clinical Laboratory Standards Institute (CLSI) document C-62A. 40 plasma samples from patients tested for PPGLs were analyzed in three mainstream commercially available kit methods. Agreement of test results from different kit methods was assessed by Passing-Bablok regression and Bland-Altman analysis. Recalibration using uniform calibrations was performed to identify the possible calibration difference.

Results: The performance of three mainstream commercially available LC-MS/MS kit methods was satisfied. The mean deviations ranges were as follows: epinephrine(E), -41.1% ~ -2.4%; norepinephrine (NE), -34.3% ~ -19.1%; metanephrine (MN), -6.5% ~ 0.3% and normetanephrine (NMN), -2.3% to 0.0%. After recalibration, the mean deviation ranges of the E and NE were reduced remarkably (E: -18% to -2.4%, NE: -19.1% to 8.0%).

Conclusions: For MN and NMN, the consistency between the three commercially available kit methods is acceptable. The results for the E and NE showed unsatisfying comparability for almost all method pairs. The application of uniform calibrators could improve consistency among the mainstream systems for the measurement of plasma catecholamines.

Objective: HbA1c values in patients with Hb Toranomon, a hemoglobin variant, are falsely high when measured with the Tosoh high-performance liquid chromatography (HPLC) analyzer. Moreover, a phenomenon has been observed in which the valley between HbA1c and HbA0 [slow valley (SV)] appears shallow on the chromatograms measured by the Tosoh HPLC analyzer in patients with Hb Toranomon. In the present study, the usefulness of the indicator, HbA1c (A1c)-adjusted SV height rate (AaSVHR), as a screening test for Hb Toranomon to quantify the SV height (SVH) was investigated.

Methods: This study included six patients with Hb Toranomon and 100 control participants, comprising 50 diabetic patients and 50 non-diabetic participants. The SVH between HbA1c and HbA0 and the height of HbA1c (A1cH) on HPLC (G9) chromatograms were measured to determine the ratio of SVH to A1cH [SVH ratio (SVHR)]. Given that the SVHR obtained showed a significant negative correlation with HbA1c measured by HPLC, SVHR was corrected with HbA1c [A1c-adjusted SVHR (AaSVHR)], but it showed no significant correlation with HbA1c.

Results: AaSVHR exceeded 14% across all participants with Hb Toranomon, but it was no more than 14% in all control participants.

Conclusion: AaSVHR measurements are useful as an Hb Toranomon screening test in patients whose HbA1c measured with a Tosoh HPLC analyzer is falsely high. A high AaSVHR exceeding 14% raises a strong suspicion for the presence of a hemoglobin variant including Hb Toranomon.

Objective: We evaluated two SDS-based preparation methods for direct identification of yeasts using MALDI-TOF MS analysis and antifungal susceptibility testing (AFST).

Methods: A total of 149 residual monomicrobial blood culture bottles (BCs) were included. For direct identification via MALDI-TOF MS analysis from positive BCs, two in-house methods with or without a 3-hr incubation step were evaluated. 55 samples were also prepared using a Sepsityper kit. In addition, to investigate the effects of differences in Candida spp. concentrations, simulated samples were incubated and colonies were counted. Direct AFST was performed using Sensititre YeastOne.

Results: Correct identification rate was 65.8% for the saponin+0.5% SDS method and 70.5% for the 20% SDS after 3-hr incubation, respectively. For the 55 samples evaluated using the Sepsityper kit, both in-house methods showed significantly higher identification rates. For the simulated samples, Candida albicans had the lowest colony counts at the time of the positive signal in the BCs. Essential agreement was above 90% for all cases except for itraconazole using the pellet prepared by the 20% SDS method. Categorical agreement was above 90% for all cases.

Conclusion: Our newly developed preparation method using 20% SDS demonstrated superior performance to a commercial kit, and can therefore be used for direct identification and AFST.

Objective: This study aimed to ascertain serum bile acid levels and explore the diagnostic value of differential bile acids in patients with intrahepatic cholestasis of pregnancy (ICP).

Methods: 94 cases of ICP patients admitted to Wenzhou People's Hospital from Oct 2020 to July 2023 were selected as the ICP group, and 184 women who underwent normal prenatal examination in the same period as the healthy control group. The baseline data and 15 kinds of serum bile acid spectrum characteristics of the two groups of pregnant women were compared, and the risk factors of serum bile acid spectrum causing ICP were analyzed by univariate and multivariate logistic regression. A logistic regression model was constructed based on significant risk factors, and ROC curve was drawn to evaluate the diagnostic effect of the model.

Results: Compared with the healthy control group, the other 14 serum bile acids except LCA were highly expressed in the ICP group, and there was significant difference (p<0.05). Univariate analysis showed that 15 serum bile acids were statistically different from ICP (p<0.05). Multivariate analysis showed that both TCA and DCA were risk factors for ICP, with 95% CI of 1.010 (1.003-1.021) and 1.003 (1.000-1.007), respectively. ROC curve showed that TCA, DCA, and TCA+ DCA were the diagnostic criteria of ICP, and their maximum diagnostic value was TCA+ DCA (AUC 0.932). The AUC values of TCA and DCA were 0.929 and 0.615, respectively. TCA+ DCA could play a complementary role in the diagnosis of ICP, and its sensitivity and specificity were 90.7% and 88.9%, respectively.

Conclusion: TCA and DCA indicators are closely related to the occurrence of ICP, and the combined detection of TCA and DCA can significantly improve the efficiency of ICP diagnosis.

Objective: This study utilized data from two hospitals to investigate the relationship between the preoperative serum alanine aminotransferase to aspartate aminotransferase ratio (LSR) combined with preoperative serum tumor markers (TM) and the prognosis of patients with stage III colorectal cancer (CRC).

Methods: An analysis was performed on the clinical data of 211 patients diagnosed with stage III colorectal cancer (CRC), treated surgically at the Department of Gastrointestinal Surgery of the Second Affiliated Hospital of Nanchang University and the General Surgery Department of the First Affiliated Hospital of Nanchang University between January 2016 and December 2016. Laboratory data were also compiled for these patients. The main follow-up indicators were the five-year disease-free survival (DFS) and the five-year overall survival (OS). Survival analysis was carried out using the log-rank test, and Cox regression analysis was applied to ascertain independent prognostic factors for survival. Rank sum tests were utilized to compare categorical variables, with a p-value of less than 0.05 considered statistically significant.

Result: Between January 2016 and December 2016, a total of 211 patients were enrolled in the two hospitals, including 113 males and 98 females. Multivariate analysis showed that the independent prognostic factors for five-year disease-free survival of stage III colorectal cancer were LSR (HR: 0.45,95% CI:0.31-0.67, p<0.001) and CEA (HR: 2.11,95%CI:1.42-3.13, p<0.001). Independent prognostic factors for five-year overall survival of stage III colorectal cancer were LSR (HR:0.48, 95%CI:0.31-0.75, p=0.001) and CA199 (HR:1.73, 95%CI:1.11-2.68, p=0.015). Kaplan-Meier survival analysis showed that LSR combined with TM could evaluate the five-year DFS and five-year OS of stage III colorectal cancer (DFS, p<0.001, OS: p=0.002).

Conclusion: 1. The prognosis of patients with high preoperative LSR and low CEA, CA199 in perioperative period is better than that of other stage III colorectal cancer patients. 2. Perioperative preoperative serum transaminase ratio combined with CEA, CA199 has a certain reference value in the prognosis assessment of stage III colorectal cancer.

Objective: We explored the mechanism of action of microRNA-146a in the lung cancer cell line A549 through the Noth1/Hes-1 signal pathway.

Methods: A549 cells were divided into a control NT group without transfection, NC group with transfection of miR-146a-NC, and MM group with transfection of miR-146a mimics. We employed qRT-PCR, MTT, Hoechst33258 fluorescence staining, Transwell assay, cell wound scratch assay, and Western blot for detection of the expression of miR-146a and Notch1/Hes-1, cell activity, apoptotic capacity, and invasion and migration abilities.

Results: The expression of miR-146a in MM group was the highest, illustrating transfection was successful. The expression of Notch1 and Hes-1 mRNA in MM group was reduced compared to in the NC and NT groups (P<0.05). The activity of Notch1 and Hes-1 were reduced by transfection of miR-146a as documented by the double luciferase reporter gene assay (P<0.05). The cellular activity in the MM group was lowest after the 48 hours of culture. There was a significant difference compared with the other two groups (P<0.05). Cell activity in the three groups after the 96 hours of culture was higher than after the 72 hours and 48 hours of culture equally (P<0.05).

Conclusion: The low expression of miR-146a could have tumor-prompting action on human NSCLC cells. Up-regulation of the expression of miR-146a restrained biological behaviors such as cell apoptosis and migration through targeted regulation involving the signal pathway of Noth1/Hes-1.

Objective: Pancreatic adenocarcinoma (PAAD) is an aggressive and lethal cancer with limited treatment options and poor prognosis. Monoacylglycerol lipase (MGLL), a key enzyme in lipid metabolism, is thought to play a role in tumor progression, but its function in PAAD remains unclear. This study aims to investigate the expression and functional significance of MGLL in pancreatic cancer and evaluate melatonin (MLT) as a potential inhibitor of MGLL.

Methods: Tissue samples from 22 PAAD patients were collected and analyzed by immunohistochemistry to assess MGLL expression. Bioinformatics tools, including GEPIA and the Human Protein Atlas, were used to compare MGLL expression in pancreatic cancer versus normal tissue, and survival analysis was conducted. In vitro assays, including CCK-8, colony formation, wound healing, and Transwell, were performed on PANC-1 cells to explore the effects of MGLL knockdown using siRNA. Western blotting was used to analyze the impact of MGLL on the P-AKT/AKT signaling pathway and MMP-2/9 expression. The effect of MLT on MGLL expression was evaluated using RNA-seq and qRT-PCR.

Results: MGLL was significantly overexpressed in PAAD tissues and was associated with poor patient prognosis. Silencing MGLL in PANC-1 cells reduced cell proliferation, migration, and invasion, likely via the P-AKT/AKT pathway. MLT treatment effectively downregulated MGLL expression, as well as MMP-2 and MMP-9, suggesting that MLT could serve as a potential MGLL inhibitor.

Conclusion: MGLL promotes pancreatic cancer progression through the P-AKT/AKT signaling pathway, and MLT may offer a novel therapeutic strategy by inhibiting MGLL expression. These findings highlight MGLL as a potential therapeutic target for PAAD.