1.The level of autophagy induced by oxygen glucose deprivation/reperfusion injury in L02 cell
Acta Universitatis Medicinalis Anhui 2017;52(4):475-479
Objective To explore the level of autophagy induced by oxygen glucose deprivation/reperfusion(OGD/R) injury in L02 cell.Methods L02 cells were cultured to establish the model of OGD/R injury and simulate clinical hepatic ischemia-reperfusion injury.The L02 cells were randomly divided into 5 groups : normal control group, oxygen-glucose deprivation 6 h/reperfusion 1,3,6,12 h group (OGD 6 h/R 1,3,6,12 h).Then observe the form changes of the L02 cells by optical microscope.The appreciation of the company's relative L02 cells was detected by MTT.The expression of autophagy related proteins such as Beclin-1, LC3 and p62 were evaluated by Western blot.Results Compared with the normal control group, the form damaged and the cells proliferation activity of L02 cells in the OGD/R group were gradually increased in a time-dependent manner.Compared with the normal control group, autophagy related proteins LC3 , Beclin-1 were increased at OGD 6 h/R 1 h.The expression of LC3 was gradually increased as the time went on and was increased gradually at OGD 6 h/R 6 h, reached a peak at OGD 6 h/R 12 h(P<0.01).The expression of Beclin-1 was gradually increased as the time went on and was increased gradually at OGD 6 h/R 6 h and OGD 6 h/R 12 h (P<0.01).The expression of p62 had no obvious change at OGD 6 h/R 1 h and OGD 6 h/R 3 h, began to increase sharply at OGD 6 h/R 6 h and reached a peak at OGD 6 h/R 12 h(P<0.01).Conclusion Our data suggests that oxygen-glucose deprivation/reperfusion may increase the level of autophagy and lead to autophagic cell death in L02 cell.
2.Preliminary study of radiation dosage in the lower limb factures surgery with single source dual energy CT
Sensen YAN ; Yang LI ; Min LUO ; Xiuya CEN
Chinese Journal of Primary Medicine and Pharmacy 2015;22(4):500-503,641
Objective To assess the clinical application value of single-source dual energy CT(SSDE),by measuring the multi-slice CT(MSCT) and single-source dual energy CT(SSDE) lower limb fractures image noise and contrast noise ratio(CNR),comparing the difference in the radiation dosage.Methods 39 cases of lower extremity fractures by clinical diagnosis were included in the study.Preoperation or postoperation the patients underwent MSCT scans,postoperation or postoperative follow-up SSDE CT scans were conducted,two sets of images were achieved.We measured and compared conventional CT group with 130 key energy spectrum group (prior studies showed the elimination of metal artifacts,images,subjective scoring the highest photon energy level) no significant artifacts cancellous bone area,compared the image background noise and contrast noise ratio (CNR),and radiation dosage,using paired sample t-test.Results Two sets of data for the same patient before and after surgery or after review of images,which body mass index (BMI) was negligible.The CTDIvol value of the conventional MSCT was (7.94 ± 1.69) mGy,the dual-energy CT was (7.04 ± 0.97) mGy,with significant difference (t =2.71,P < 0.05) ; The image noise of the conventional MSCT was (44.36 ± 15.66) Hu,the dual-energy CT was (15.04 ± 8.23) Hu,with significant difference (t =15.26,P < 0.05) ; The CNR of the conventional MSCT was (3.96 ± 1.78),the dual-energy CT was (4.62 ± 2.80),with significant difference (t =-2.14,P < 0.05).Conclusion Compared with conventional MSCT,single source dual-energy CT imaging provides improved image CNR,obviously eliminate metal artifacts,to better show the complex structure for lower limb fracture surgery,and reduction in radiation dosage.
3.Effect of propofol on autophagy during oxygen-glucose deprivation/restoration in human liver cells
Xiuya MA ; Yujun SHEN ; Yuanhai LI ; Yuxian SHEN
Chinese Journal of Anesthesiology 2016;36(6):725-727
Objective To evaluate the effect of propofol on autophagy during oxygen-glucose deprivation and restoration (OGD/R) in human liver cells.Methods Human hepatic HL-7702 cells at the logarithmic growth phase were seeded into culture plates and randomly divided into 3 groups (n =12 each) using a random number table:control group (group C),OGD/R group,and propofol + OGD/R group (group P+OGD/R).The cells were cultured in normal culture medium in group C.In OGD/R and P+OGD/R groups,the cells were subjected to O2-glucose deprivation for 6 h followed by restoration of O2-glucose supply for 12 h.Propofol with a final concentration of 50 mmol/L was added at 10 min before oxygen-glucose deprivation.The cell viability was detected by MTT assay.The expression of autophagy-related proteins such as microtubule-associated protein light chain 3 (LC3) and Beclin-1 was evaluated by Western blot.Immunofluorescence was used to determine the number and distribution of autophagosomes.Results Compared with group C,the cell viability was significantly decreased,the expression of LC3 and Beclin-1 was significantly up-regulated (P<0.05),and the number of autophagosomes was significantly increased in OGD/R and P+OGD/R groups.Compared with group OGD/R,the cell viability was significantly increased,the expression of LC3 and Beclin-1 was significantly down-regulated (P<0.05),and the number of autophagosomes was significantly decreased in group P+OGD/R.Conclusion The mechanism by which propofol reduces OGD/R injury is probably related to inhibition of autophagy in human liver cells.
4.De Novo and Inherited SETD1A Variants in Early-onset Epilepsy.
Xiuya YU ; Lin YANG ; Jin LI ; Wanxing LI ; Dongzhi LI ; Ran WANG ; Kai WU ; Wenhao CHEN ; Yi ZHANG ; Zilong QIU ; Wenhao ZHOU
Neuroscience Bulletin 2019;35(6):1045-1057
Early-onset epilepsy is a neurological abnormality in childhood, and it is especially common in the first 2 years after birth. Seizures in early life mostly result from structural or metabolic disorders in the brain, and the genetic causes of idiopathic seizures have been extensively investigated. In this study, we identified four missense mutations in the SETD1A gene (SET domain-containing 1A, histone lysine methyltransferase): three de novo mutations in three individuals and one inherited mutation in a four-generation family. Whole-exome sequencing indicated that all four of these mutations were responsible for the seizures. Mutations of SETD1A have been implicated in schizophrenia and developmental disorders, so we examined the role of the four mutations (R913C, Q269R, G1369R, and R1392H) in neural development. We found that their expression in mouse primary cortical neurons affected excitatory synapse development. Moreover, expression of the R913C mutation also affected the migration of cortical neurons in the mouse brain. We further identified two common genes (Neurl4 and Usp39) affected by mutations of SETD1A. These results suggested that the mutations of SETD1A play a fundamental role in abnormal synaptic function and the development of neurons, so they may be pathogenic factors for neurodevelopmental disorders.
5.BMAL1 regulates mitochondrial fission and mitophagy through mitochondrial protein BNIP3 and is critical in the development of dilated cardiomyopathy.
Ermin LI ; Xiuya LI ; Jie HUANG ; Chen XU ; Qianqian LIANG ; Kehan REN ; Aobing BAI ; Chao LU ; Ruizhe QIAN ; Ning SUN
Protein & Cell 2020;11(9):661-679
Dysregulation of circadian rhythms associates with cardiovascular disorders. It is known that deletion of the core circadian gene Bmal1 in mice causes dilated cardiomyopathy. However, the biological rhythm regulation system in mouse is very different from that of humans. Whether BMAL1 plays a role in regulating human heart function remains unclear. Here we generated a BMAL1 knockout human embryonic stem cell (hESC) model and further derived human BMAL1 deficient cardiomyocytes. We show that BMAL1 deficient hESC-derived cardiomyocytes exhibited typical phenotypes of dilated cardiomyopathy including attenuated contractility, calcium dysregulation, and disorganized myofilaments. In addition, mitochondrial fission and mitophagy were suppressed in BMAL1 deficient hESC-cardiomyocytes, which resulted in significantly attenuated mitochondrial oxidative phosphorylation and compromised cardiomyocyte function. We also found that BMAL1 binds to the E-box element in the promoter region of BNIP3 gene and specifically controls BNIP3 protein expression. BMAL1 knockout directly reduced BNIP3 protein level, causing compromised mitophagy and mitochondria dysfunction and thereby leading to compromised cardiomyocyte function. Our data indicated that the core circadian gene BMAL1 is critical for normal mitochondria activities and cardiac function. Circadian rhythm disruption may directly link to compromised heart function and dilated cardiomyopathy in humans.
6.The roles of Mesp family proteins: functional diversity and redundancy in differentiation of pluripotent stem cells and mammalian mesodermal development.
Qianqian LIANG ; Chen XU ; Xinyun CHEN ; Xiuya LI ; Chao LU ; Ping ZHOU ; Lianhua YIN ; Ruizhe QIAN ; Sifeng CHEN ; Zhendong LING ; Ning SUN
Protein & Cell 2015;6(8):553-561
Mesp family proteins comprise two members named mesodermal posterior 1 (Mesp1) and mesodermal posterior 2 (Mesp2). Both Mesp1 and Mesp2 are transcription factors and they share an almost identical basic helix-loop-helix motif. They have been shown to play critical regulating roles in mammalian heart and somite development. Mesp1 sits in the core of the complicated regulatory network for generation of cardiovascular progenitors while Mesp2 is central for somitogenesis. Here we summarize the similarities and differences in their molecular functions during mammalian early mesodermal development and discuss possible future research directions for further study of the functions of Mesp1 and Mesp2. A comprehensive knowledge of molecular functions of Mesp family proteins will eventually help us better understand mammalian heart development and somitogenesis as well as improve the production of specific cell types from pluripotent stem cells for future regenerative therapies.
Amino Acid Sequence
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Animals
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Basic Helix-Loop-Helix Transcription Factors
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genetics
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Cell Differentiation
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genetics
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Gene Expression Regulation, Developmental
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Mesoderm
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embryology
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metabolism
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Mice, Knockout
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Molecular Sequence Data
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Pluripotent Stem Cells
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metabolism
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Sequence Homology, Amino Acid