1.Role and mechanism of mitochondrial calcium uniporter in the cytoskeleton of pancreatic ductal epithelial cells in a mouse model of acute pancreatitis
Qiaofeng CHEN ; Qingzi FU ; Huiying YANG ; Junbo HONG ; Liang ZHU ; Zhenzhen YANG ; Guodu TANG ; Shiyu ZHANG
Journal of Clinical Hepatology 2026;42(2):400-408
ObjectiveTo investigate the effect of mitochondrial calcium uniporter (MCU) on the cytoskeleton of pancreatic ductal epithelial cells in a mouse model of acute pancreatitis (AP) induced by caerulein (CAE), to analyze the role of MCU in the development of AP, and to provide a theoretical basis for clinical treatment. MethodsIn the in vivo experiment, wild-type male C57BL6/J mice, aged 4 weeks, were randomly divided into control group and AP group, with 6 mice in each group. The mice in the AP group were given intraperitoneal injection of CAE to establish a model of AP, and those in the control group were given intraperitoneal injection of an equal volume of normal saline. Serum and pancreatic tissue samples were collected after 24 hours of modeling. HE staining was used to observe pancreatic histopathological changes; Western Blot was used to measure the expression levels of MCU, glutathione peroxidase 4 (GPX4), and acyl-CoA synthetase long chain family member 4 (ASCL4); kits were used to measure the serum level of amylase. In the in vitro experiment, the human pancreatic ductal epithelial cell line HPDE6-C7 was co-cultured with CAE for 24 hours to establish an in vitro AP model, and the cells were divided into control group, CAE group, RR (an MCU activity inhibitor) group, CAE+RR group, Fer-1 (an ferroptosis inhibitor) group, CAE+Fer-1 group, Erastin (an ferroptosis inducer) group, and CAE+Erastin group. CCK-8 assay was used to observe the influence of different agents on cell viability; Western Blot was used to measure the expression levels of MCU, GPX4, and ASCL4; immunofluorescence assay was used to measure reactive oxygen species (ROS), actin cytoskeleton, and monolayer permeability; kits were used to measure the concentrations of malondialdehyde (MDA), glutathione (GSH), Fe2+, and total iron. A one-way analysis of variance was used for comparison of continuous data between multiple groups, and the least significant difference t-test was used for comparison between two groups. ResultsIn the in vivo experiment, compared with the control group, the AP group had significant increases in pancreatic histopathological score, the serum level of amylase, and the expression levels of MCU and ASCL4, as well as a significant reduction in the expression of GPX4 (all P<0.05). In the in vitro experiment, compared with the control group, the CAE group had significant increases in the expression levels of MCU and ASCL4, a significant reduction in the expression of GPX4, and significant increases in the concentrations of Fe2+, total iron, and MDA, the green fluorescence intensity of ROS, and monolayer permeability, as well as a significant reduction in the concentration of GSH (all P<0.05), with the presence of actin cytoskeleton disruption. Compared with the CAE group, the CAE+RR group had a significant increase in the expression level of GPX4, a significant reduction in the expression level of ASCL4, and significant reductions in the concentrations of Fe2+, total iron, and MDA, the green fluorescence intensity of ROS, and monolayer permeability and a significant increase in the concentration of GSH (all P<0.05), with alleviation of actin cytoskeleton disruption. Compared with the CAE group, the CAE+Fer-1 group had significant reductions in the concentrations of Fe2+, total iron, and MDA, the green fluorescence intensity of ROS, and monolayer permeability and a significant increase in the concentration of GSH (all P<0.05), with alleviation of actin cytoskeleton disruption. Compared with the CAE group, the CAE+Erastin group had significant increases in the concentrations of Fe2+, total iron, and MDA, the green fluorescence intensity of ROS, and monolayer permeability and a significant reduction in the concentration of GSH (all P<0.05), with aggravation of actin cytoskeleton disruption. ConclusionDuring the onset of AP, MCU mediates oxidative stress-induced ferroptosis and leads to the disruption of the pancreatic ductal epithelial barrier, which may be one of the possible pathogeneses of AP.
2.Discussion on the reference range of thromboelastogram in 916 healthy adults in Shenzhen area
Weicheng LI ; Kunlun WU ; Siqi CAI ; Guodu ZHU ; Yubao ZHONG ; Yunjing XU ; Nansheng CAI ; Lili WU ; Zhenglin WU
Chinese Journal of Blood Transfusion 2022;35(3):304-307
【Objective】 To determine the reference range of thromboelastogram(TEG) and establish a TEG feature for local population by measuring TEG parameters in healthy adults in Shenzhen comparing the difference between gender and age, and analyzing the reference data provided by reagent manufacturer. 【Methods】 A total of 916 healthy adults, aged between 19 to 59, who did their regular health checks in our hospital from September 2020 to August 2021 were selected. The TEG(from Lepu Medical Technology Co., Ltd.) was performed, and the clot reaction time(R), clot formation time(K), coagulation angle(α-Angle), maximum amplitude(MA), coagulation index(CI), fibrinolysis index LY30 and the estimated percent lysis (EPL) were analyzed. 【Results】 The reference ranges of TEG parameters, including R, K, α-Angle, MA, CI, LY30 and EPL, of 916 healthy adults from Shenzhen were 3.25~8.19 min, 0.66~3.18min, 47.70~76.56deg, 50.05~72.91mm, -4.3~3.4, 0~2.2% and 0~3%, respectively. The value of α-Angle, CI, K, LY30, MA and R didn’t all meet the given range provided by the manufacturer; some were exceeding and some inferior to. A total of 227 out of 916 individuals presented abnormal results, relative to the references, in at least one parameter, and 78 were diagnosed of abnormal coagulation based on the given reference range, with a specificity of 75.2%. 【Conclusion】 The reference range of TEG parameters of Shenzhen locals is significantly different from that provided by manufacturers. And it is imperative for local TEG laboratories to establish their own reference ranges according to age and gender groups based on local population characteristics.

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