1.ADAR2 inhibits MAVS expression by editing its 3'UTR in cells
Tao LI ; Jingyuan LI ; Xilin ZHU ; Xiaopan WU ; Ying LIU
Basic & Clinical Medicine 2017;37(6):747-751
Objective To investigate the effect of ADAR2 on the expression of MAVS and its mechanism.MethodsThe RT-qRCR was used to detect the expression of ADAR gene family, the expression level of ADAR2 and MAVS in cells.The effect of ADAR2 on the 3'UTR region of MAVS was detected by Pyrosequencing.To detect the expression of luciferase by dual luciferase reporter plasmid assay;The expression of ADAR2 and MAVS were detected by Western blot.Results In the ADAR family, the abundance of ADAR1 was the highest, followed by ADAR2, but the expression of ADAR3 was poor, which was almost impossible to detect(P<0.05).ADAR2 played a critical role in RNA editing of chr20:3869744 sites on the 3'UTR region of MAVS(P<0.001).On the 3'UTR editing site of MAVS, the luciferase activity of the edited G allele was significantly lower than that of the normal A allele(P<0.01).At the level of transcription and translation, ADAR2 significantly inhibited the expression of MAVS(P<0.05).Conclusions ADAR2 by editing MAVS` 3'UTR on the chr20:3869744 site, which makes the occurrence of A to G replacement, inhibits the expression of MAVS.
2.Evaluation of Estrogenic Effects ofCoicis Semen Aqueous Extracts
Xiaoke ZHENG ; Na ZHANG ; Xiaofan WANG ; Jinli HE ; Yangang CAO ; Lingling LI ; Peipei YUAN ; Xiaopan LIU ; Yong LOU ; Pengfei LI ; Weisheng FENG
World Science and Technology-Modernization of Traditional Chinese Medicine 2014;(9):1974-1979
This study was aimed to investigate the estrogenic effects ofCoicis Semen in order to preliminarily discuss the mechanism. Mouse uterine weight test and MCF-7 cell proliferation assay were used to evaluate the estrogenic effects ofC. Semen. Reporter gene assays were adopted to explore the action mechanism ofC. Semen. In reporter gene assay, HEK293 cells were co-transfected with pERE-TAL-luc, pβgal-Control, pCXN2-hERα, or pCXN2-hERβ. And the expression of reporter gene luc was controlled by ERE. Mouse uterine weight test showed that compared with the control group, the aqueous extracts ofC. Semen can increase the uterus index of premature female mice (P<0.01). It can significantly promote the proliferation of MCF-7 cells in the medium without estrogen (P<0.01). The reporter gene controlled by ERE technology showed thatwhen mediated by ERα or ERβ respectively, the normalized luciferase activity of aqueous extracts ofC. Semen was significantly higher than activity of the control group (P< 0.05 orP< 0.01). It was concluded that the aqueous extracts ofC.Semen can increase the uterus index of premature female mice and promote the proliferation of MCF-7 cells in the medium without estrogen. We found the estrogenic effects ofC. Semen for the first time. And the estrogenic effects ofC. Semen were mainly mediated by ERβ.
3.Optimal dose of dexmedetomidine combined with propofol for anesthesia in patients undergoing modified electroconvulsive therapy
Qian HAO ; Baojiang LIU ; Jianhong LI ; Xiaopan WANG ; Li ZHOU ; Jieping LYU
Chinese Journal of Anesthesiology 2020;40(1):65-67
Objective:To investigate the optimal dose of dexmedetomidine combined with propofol for anesthesia in patients undergoing modified electroconvulsive therapy (MECT).Methods:One hundred and sixty patients of both sexes, aged 20-60 yr, weighing 45-80 kg, of American Society of Anesthesiologists physical status Ⅰ or Ⅱ, scheduled for elective MECT, were allocated into 4 groups ( n=40 each) by a random number table method: different doses of dexmedetomidine combined with propofol group (D 1, D 2 and D 3 groups) and routine anesthesia group (group C). Dexmedetomidine 0.2, 0.4 and 0.6 μg/kg were intravenously injected in D 1, D 2 and D 3 groups, respectively, the equal volume of normal saline was given instead in group C, and propofol 1.0 mg/kg and succinylcholine 0.5 mg/kg were intravenously injected in turn 10 min later.Venous blood samples were collected before giving dexmedetomidine (T 0) and at 1 min after the end of MECT (T 1) for determination of the plasma epinephrine (E) and norepinephrine (NE) concentrations.Propofol consumption, occurrence of cardiovascular events, duration of epilespsy and energy suppression index were recorded. Results:Compared with group C, the plasma E and NE concentrations were significantly decreased at T 4, and the propofol consumption was reduced in D 1, D 2 and D 3 groups ( P<0.05). Compared with group D 2, the plasma E and NE concentrations were significantly increased at T 1 in group D 1 and decreased at T 1 in group D 3 ( P<0.05). The incidence of adverse cardiovascular events was significantly increased in group D 3 than in the other 3 groups ( P<0.05). There was no significant difference in duration of epilespsy or energy suppfession index among the 4 groups( P>0.05). Conclusion:The optimal dose of dexmedetomidine combined with propofol 1.0 mg/kg is 0.4 μg/kg when used for anesthesia in the patients undergoing MECT.
4.Related factors of three-vessel disease in patients with stable coronary artery disease
Yajun ZHAO ; Xuelin CHENG ; Ming LIU ; Xiaopan LI ; Jing ZHOU ; Jian ZOU ; Yuxiang DAI ; Sunfang JIANG
Chinese Journal of General Practitioners 2023;22(4):394-398
Objective:To analyze the risk factors of three-vessel disease (TVD) in patients with stable coronary artery disease (SCAD).Methods:The clinical data of 447 patients with SCAD diagnosed in Zhongshan Hospital from May 2019 to April 2020 were retrospectively analyzed, including 108 cases with the single-vessel disease (SVD), 136 cases with the two-vessel disease, and 203 cases with three-vessel disease. The general data and hematological indexes were compared between patients with SVD and those with TVD; the related factors for TVD in SCAD patients were analyzed with univariate and multivariate logistic regression.Results:There were 244 males (78.5%) and 67 females (21.5%) with a median age of 57 years (64, 69). Univariate analysis showed that there were significant differences in diabetes history ( χ2=7.75, P=0.005), uric acid ( Z=-2.10, P=0.036), glycosylated hemoglobin ( Z=-2.77, P=0.006) and high density lipoprotein cholesterol (HDL-C) ( Z=-2.99, P=0.003) levels between SVD and TVD groups. Multivariate analysis showed that the high level of blood uric acid ( OR=1.01, 95% CI: 1.00-1.01, P<0.05) and the low level of HDL-C ( OR=3.29, 95% CI:1.23-8.85, P<0.05) were related risk factors of TVD. Conclusion:High blood uric acid level and low HDL-C level are related factors for TVD in patients with SCAD.
5.Molecular mechanism of ATF6 regulating the reproduction related gene HSPA1L
Yuanyuan WANG ; Xilin ZHU ; Xiaopan WU ; Ying LIU
Basic & Clinical Medicine 2024;44(1):37-42
Objective To explore the effect of endoplasmic reticulum stress activating transcription factor 6(ATF6)on the expression of reproduction related gene heat shock protein family A member 1 like(HSPA1L)and preliminari-ly clarify its regulatory molecular mechanism.Methods The ATF6 over-expression plasmid was transfected into HEK-293T cells and the over-expression efficiency was verified by RT-qPCR and Western blot.The transcriptome sequen-cing information of testis tissue of male ATF6 knockout mice was used to screen five reproduction related genes down-stream of ATF6.The dual luciferase reporter assay selected the downstream genes with high promoter activity and de-tected the effect of over-expression of ATF6 on the promoter activity of downstream genes.The possible binding sites of ATF6 and downstream gene promoters were predicted by gene-regulation.RT-qPCR and Western blot were used to detect the effect of over-expression of ATF6 on downstream gene expression in HEK-293T cells.Whether ATF6 binds to downstream gene promoters was determined by electrophoretic mobility shift assay(EMSA).Results The expres-sion of ATF6 mRNA(P<0.001)and protein(P<0.001 and P<0.05)in HEK-293T cells was significantly increased after transfection.HSPA1L(P<0.001 and P<0.05),a reproductive related gene downstream of ATF6 was screened by transcriptome sequencing and dual luciferase reporter assay.ATF6 promoted the truncated promoter activity of HSPA1L(P<0.001).After over-expression of ATF6,the expression of HSPA1L was significantly increased(P<0.001).The differences were statistically significant.ATF6 protein could bind to the aagtcgtcac DNA sequence of HSPA1L promoter.Conclusions ATF6,a key molecule of endoplasmic reticulum stress,regulates the expression of reproduction related gene HSPA1L by binding to the promoter of HSPA1L.This result will lay a foundation for further research on the prevention or treatment of endoplasmic reticulum stress(ERS)related male infertility.