1.Effect of erythropoietin on Bcl-xl expression in hippocampal CA1 subregion and cognizing function in rats with cerebral ischemia-reperfusion
Xiandong MENG ; Xiaohong LUO ; Xiaodan JIANG
Journal of Clinical Neurology 1997;0(06):-
Objective To explore the effect of erythropoietin(EPO) on Bcl-xl expression in hippocampal CA1 subregion and cognizing function in rats with cerebral ischemia-reperfusion(IR).Methods Male SD rats were randomly separated into sham-operated group,IR group and EPO group,and the IR models were made.At 3 h before IR models made,rHu-EPO and saline were injected into rat lateral ventricles in EPO group and IR group,respectively,with help of stereotaxic coordinates.Bcl-xl protein expression of Hippocampal CA1 subregion was detected by immunohistochemistry at 24 h after operation.The learning and memory abilities were examined by electricity maze at 4 weeks after operation.Results Compared to sham-operated group,Bcl-xl protein expression in EPO group and IR group were obviously decreased,but EPO group was significantly increased than IR group(all P
2.The protective effect of EPO on Alzheimer disease rat learning and memory injury and its effect on Bcl-xl protein expression in hippocampal CA1 subregion
Xiandong MENG ; Rong WANG ; Xiaodan JIANG ; Junpao CHEN
Chinese Journal of Behavioral Medicine and Brain Science 2010;19(8):689-691
Objective To explore the protecting mechanism of erythropoietin (EPO) on learning and memory of Alzheimer disease (AD) rats. Methods The AD model was made by injected into rat hippocampal CA1 subregion with amyloid beta-protein(Aβ). Male Spraque-Dawley rats were as the experimental objects, which were randomly separated into 3 groups including Sham, Saline control and EPO treatment. After Aβ was injected into rats hippocampal CA1 subregion ,saline or EPO was respectively injected into the lateral ventricle of rats,with help of stereotaxic coordinates, upon the designed conditions. Hippocampal CA1 subregion Bcl-xl expression changes were observed 24 hours after the operation, and learning and memory abilities were checked 4 weeks after the operation. Results 24 hours after the operation Bcl-xl expression in the EPO group and the Saline group was less than the Sham control ,while Bcl-xl positive cells( 100.42 ± 12.43/field) in the EPO group were more than in the Saline group( 82.06 ± 19.68/field ) (P < 0. 05 ). 4 weeks after the operation learning ability in the EPO group ( 20.38 ± 5.88 ) was better than Saline group ( 25.50 - 3.25 ) (P < 0. 05 ), and memory ability in the EPO group (4.75 ± 1.75 ) was better than the Saline group(2.88 ± 1.55 )(P < 0.05 ). Conclusion EPO could improve the learning and memory abilities in the model rats,and it could be related with EPO restraining Bcl-xl expression decreasing.
3.The predictive value of the expression of EZH2 on B lymphocyte subsets on the prognosis of septic patients
Dongyang ZHAO ; Zhe LI ; Xiaowei BAO ; Hong SUN ; Qian YANG ; Huijuan REN ; Xiandong LIU ; Sen JIANG ; Xiaohui ZHOU ; Lunxian TANG
Chinese Journal of Emergency Medicine 2021;30(11):1340-1348
Objective:To investigate the dynamic expression of histone methyltransferase (enhance of zeste homolog 2, EZH2) in peripheral blood B lymphocytes (CD19 +B) and memory B lymphocytes (CD19 +CD27 +B) of septic patients and its value in predicting prognosis in sepsis. Methods:From June 2018 to January 2020, 48 septic patients in the Intensive Care Unit of Shanghai East Hospital were enrolled, and 40 healthy adult volunteers were recruited as healthy controls. Septic patients were divided into the non-survivors (18 cases) and the survivors (30 cases) according to whether the patients survived at 28 days. Blood samples were collected at day 1, 3 and 7, blood routine, IL-6 and blood gas analysis were collected, and SOFA and APACHE Ⅱ scores were counted. Flow cytometry was used to detect the positive rate and the mean fluorescence intensity of EZH2 on CD19 +B lymphocytes, and the positive rate of EZH2 on CD19 +CD27 +B lymphocytes at different time points. In the healthy controls, fasting was taken only once in the morning. ROC curve was drawn and the area under the curve (AUC) was calculated to evaluate the value of expression of EZH2 on CD19 +B lymphocytes and CD19 +CD27 +B lymphocytes in predicting the prognosis of septic patients. Results:(1) Compared with the healthy controls, the positive rate and average fluorescence intensity of EZH2 on CD19 +B lymphocytes and the positive rate of EZH2 expression on CD19 +CD27 +B lymphocytes were significantly increased at day 1, 2 and 3 in septic patients ( P<0.05). Over time, the expression of EZH2 in CD19 +B lymphocytes and CD19 +CD27 +B lymphocytes increased gradually ( P<0.05). (2) Compared with the survivors, the positive rate of EZH2 on CD19 +B lymphocytes of the non-survivors was increased at day 1, but the positive rate of EZH2 on CD19 +CD27 +B lymphocytes of the non-survivors was decreased at day 3 and 7 ( P<0.05). (3) The positive rate of EZH2 on CD19 +B lymphocytes, APACHE Ⅱ score, SOFA score and IL-6 level in septic patients at day 1 were independently associated with 28-day mortality. (4) The AUC of APACHEⅡ score was 0.907 (95% CI: 0.825-0.990), and the sensitivity and the specificity were 88.89% and 76.67%. The AUC of SOFA score was 0.831 (95% CI: 0.706-0.955), and the sensitivity and the specificity was 66.67% and 86.67%; The AUC of EZH2 positive rate on CD19 +B lymphocytes were 0.799 (95% CI: 0.657-0.941), and the sensitivity and specificity were 88.89% and 80.77%, respectively, the sensitivity was better than SOFA score, and the specificity was higher than APACHEⅡ score. Conclusions:The high expression of EZH2 on B lymphocytes in septic patients is associated with poor prognosis. Dynamic monitoring of EZH2 expression on B lymphocytes has certain predictive value for sepsis.
4.Fipronil and its metabolites in indoor dust in Shenzhen in 2020 -2021: analysis of residual levels and temporal variations
Ying JIANG ; Xiandong ZENG ; Jie LIU ; Shiyi FANG ; Zhouning FANG ; Jianhui YUAN
Journal of Public Health and Preventive Medicine 2023;34(2):27-31
Objective To explore the residual level of FPMs in indoor dust samples in Shenzhen from 2020 and 2021, and to analyze its temporal distribution characteristics. Methods In the present study, indoor dust samples (n=193) from residential buildings in Shenzhen. were collected to analyze the temporal variation characteristics of FPMs. Ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) was applied to determine the concentrations of FP and its four major metabolites (fipronil-sulfone, fipronil-sulfide, fipronil-desulfinyl, and fipronil-amide; abbreviated as FP-SFO, FP-SFI, FP-DES, and FP-AM) in the samples. The sum of the concentrations of FP and its four metabolites was represented as additive mass concentration (ΣFPMs). Additionaly, Wilcoxon test was performed to determine the temporal distribution differences of FPMs’ concentrations. Results From 2020 to 2021, the concentration of ΣFPMs for the in door dust samples in Shenzhen ranged from 0.51 to 4 415 ng/g (median: 18.8ng/g). FP, FP-SFO AND FP-SFI were the major target analytes in the sample with detection rates of 90.60%,86.20% and 75.40%, respectively. The detection rates of other metabolites were low (≤ 44.3%). Analysis of the temporal variation trend of FPMs’ concentrations showed that there was no significant difference in the levels of ΣFPMs between warm season(spring and summer)and cold season(autumn and winter)in the indoor dust samples from 2020 to 2021(2.38 vs 2.84ng/g , P > 0.05). However , the concentrations of FP-SFI and ΣFPMs in the indoor dust samples collected from 2021 showed an significantly increasing trend compared with 2020(1.02 vs 1.89 , 17.80vs. 20.10 ng/g , P < 0.05). Conclusion From 2020 to 2021 , the detection level of FPMs in indoor dust in Shenzhen is relatively high and shows an upward trend , with no obvious seasonal difference. However, whether the residual level of FPMs in indoor dust poses a risk to human health needs further study.
5.Effect and mechanism of Erastin on ferroptosis of acute myeloid leukemia cells
Xiandong Jiang ; Yingying Huang ; Xiaoying Hong ; Xindi Lin ; Donghong Lin ; Liping Lin
Acta Universitatis Medicinalis Anhui 2025;60(1):15-21
Objective :
To explore the role of lysophosphatidylcholine acyltransferase 3(LPCAT3) in Erastin-induced ferroptosis of acute myeloid leukemia(AML) cells and its related molecular regulatory mechanisms.
Methods :
Tetrazolium salt(MTS) method was used to detect the sensitivity of different AML cells to the classic ferroptosis inducer Erastin, real time quantitative polymerase chain reaction(qPCR) was used to detect the basal expression level ofLPCAT3mRNA, and the correlation between them was analyzed. Lentivirus-mediatedLPCAT3overexpression AML cell lines(OE group) and negative control lines(NC group) were constructed. After Erastin intervention, MTS, flow cytometry, and micromethods were used to detect cell viability, lipid reactive oxygen species(ROS), and Malondialdehyde(MDA), respectively. qPCR and Western blot were used to detect unfolded protein response(UPR) classic pathway signaling molecules(PERK, ATF4, GRP78, etc.) expression levels. The above ferroptosis-related indicators were detected after combined intervention with the UPR inhibitor 4-phenylbutyric acid(4-PBA), and the regulatory relationship was analyzed.
Results :
Four different types of AML cells had different sensitivities to ferroptosis, among which K562 cells were relatively insensitive. The IC50of the four types of AML cells to Erastin was negatively correlated with the expression level ofLPCAT3(r=-0.919,P<0.001). After Erastin intervention, the cell viability of K562 cells in the OE group was significantly inhibited by Erastin compared with the NC group(P<0.001), and the levels of lipid ROS and MDA increased(P<0.001). The results of qPCR and Western blot showed that, compared with the NC group, the mRNA and protein expression of UPR classic pathway moleculesPERK,ATF4, andGRP78mRNA and protein increased in the OE group(P<0.01). After inhibiting the UPR pathway by 4-PBA, the viability of K562 cells decreased(P<0.01), and lipid ROS and MDA levels increased(P<0.01) compared with the uninhibited state.
Conclusion
Overexpression ofLPCAT3can promote ferroptosis in K562 cells, and this process is negatively regulated by the classical UPR pathway PERK/ATF.