1.Induction of apoptosis in PC12 cells by homocysteine
Jinxiu CAO ; Binghua WANG ; Yun WANG ; Lida CHEN ; Wenjing ZOU ; Ying ZHENG
Chinese Journal of Pathophysiology 1986;0(03):-
AIM: To explore the effects of homocysteine on the apoptosis in PC12 cells and the relationship between the apoptosis and the expression of the bcl-2 as well as bax gene. METHODS: Cell viability was determined by MTT assay. Cell apoptosis was assessed by phase-contrast microscope, fluorescence microscopy and flow cytometry (FCM). The expression of bcl-2 and bax mRNA was measured by semiquantitative reverse transcription polymerse chain reaction (RT-PCR). RESULTS: Treatment of PC12 cells with Hcy plus 10 ?mol/L copper for 12 h, in the range of 0.125, 0.25, 0.5, 1.0 mmol/L, caused a great decrease in cell viability: 81%, 79%, 69%, 57%, and induced typical morphological changes of apoptosis in a dose-dependent manner. The apoptosis ratios were respectively 8.00%, 10.37%, 17.26% and 20.19%, respectively. The expression of bcl-2 was significantly decreased (from 0.517 to 0.198) whereas bax was significantly increased (from 0.302 to 0.619). CONCLUSION: Homocysteine plus copper may induce apoptosis in PC12 cells through the down-regulation of bcl-2 and the up-regulation of bax gene expression.
2.Clinical features and genetic analysis of a Chinese pedigree affected with X-linked adrenoleukodystrophy.
Lingen KONG ; Jinhua QIU ; Caiming LI ; Qianwu ZHOU ; Zhiwei QIU ; Shunzhi ZHUANG ; Jinxiu ZOU ; Yanxia ZHENG
Chinese Journal of Medical Genetics 2022;39(1):60-63
OBJECTIVE:
To analyze the clinical features and variants of ABCD1 gene in a Chinese pedigree affected with X-linked adrenoleukodystrophy.
METHODS:
Clinical data of the proband were collected and analyzed. Potential variant of the ABCD1 gene were analyzed by PCR and Sanger sequencing of the proband, his parents and 100 unrelated healthy individuals.
RESULTS:
The prominent features of the proband included cerebellar and brainstem lesions, along with increased serum level of very-long chain fatty acids. He was found to harbor a hemizygous c.1509delG (p.L504Sfs*54) variant of the ABCD1 gene, for which his mother was heterozygous. The same variant was not detected in his father and 100 healthy controls.
CONCLUSION
X-linked adrenoleukodystrophy has a variety of clinical manifestations. Discovery of the c.1509delG (p.L504Sfs*54), as a novel pathogenic variant of the ABCD1 gene, has enabled diagnosis and genetic counseling for this pedigree.
Adrenoleukodystrophy/genetics*
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Asians/genetics*
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China
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Female
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Genetic Testing
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Humans
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Male
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Mutation
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Pedigree
3.Effect of complement fragment C3a on the phenotype of mice podocytes in vitro
Jinxiu DENG ; Caiming CHEN ; Luli ZHENG ; Jiong CUI ; Yanfang XU ; Zhenhuan ZOU ; Zhenzhou LI ; Jianxin WAN
Chinese Journal of Nephrology 2018;34(2):106-114
Objective To investigate the effect of complement C3a on mouse podocytes phenotype transformation.Methods Purified C3a recombinant protein was used to stimulate mature mouse podocytes.The expression of the mature podocyte markers synaptopodin,podocin,nephrin,CD2-associated protein (CD2AP) and the mesenchymal cell markers fibroblast specific protein 1 (FSP-1),α-smooth muscle actin (α-SMA) were detected by RT-PCR,Western blotting,immunochemistry and immunofluorescence,respectively.Some podocytes were transfected with integrin-linked kinase (ILK) siRNA before the administration of C3a,the expression of nephrin and α-SMA were accessed by Western blotting,and the expression of Snail and α-actinin 4 were accessed by Western blotting and immunochemical method.The migration ability of podocytes was observed by scratch test.Results Immunocytochemistry and immunofluorescence analysis showed that synaptopodin,podocin,nephrin,CD2AP were highly expressed by mature mouse podocytes.The expression of these podocyte markers could be markedly inhibited after 24 h of C3a (0.1 μmol/L) treatment,and accompanied by the induction of mesenchymal markers FSP-1 and α-SMA.Compared with control group,the mRNA levels of synaptopodin,podocin,CD2AP and nephrin were significantly repressed by the administration of C3a in a dose-dependent manner,whereas the transcription of FSP-1 and α-SMA were remarkably up-regulated by C3a treatment (P < 0.05,respectively).Western blotting analysis also confirmed the decrease of synaptopodin,podocin,nephrin and CD2AP protein and the increase of FSP-1 and α-SMA protein were closely depend on the C3a concentration (P < 0.05,respectively).To further assess the downstream of C3a,some podocytes were transfected with ILK siRNA before the administration of C3a.Compared with C3a group,the protein levels of nephrin and α-SMA were significantly changed by the administration of ILK siRNA (P < 0.05,respectively).The expression of α-actinin 4 and Snail induced by C3a were inhibited by ILK knockdown (P < 0.05,respectively),accompanied by a decline of cell migration potency.Conclusion Complement fragment C3a can induce transformation of mouse podocytes to mesenehymal cells,and ILK signaling pathway is involved in this cell type transformation.
4.Preliminary analysis of qualitative changes in suspended red blood cells under different low-pressure and hypoxic conditions
Xuelian YONG ; Li ZOU ; Lei LIU ; Qian ZANG ; Jinxiu JI ; Si CHENG ; Yongqiang LI
Chinese Journal of Blood Transfusion 2023;36(1):15-19
【Objective】 To evaluate the quality of suspended red blood cells (SRBC) under low-pressure and hypoxic conditions, in order to lay a theoretical foundation for airline delivery, air drop and storage of SRBC on plateau. 【Methods】 The low-pressure and hypoxic conditions (0.026 MPa and 0.047 MPa) were simulated by a pressure control device. SRBC were divided into 3 groups (5 bags/group, 1.5 U/bag). Each group was stored in 0.026 MPa environment at 2℃-6℃ for 24 hours (named 0.026 MPa), in 0.047MPa environment at 2℃-6℃ for 7 days (named 0.047 MPa), and in a blood storage refrigerator at 4℃ (named the control), respectively. The storage cell characteristics were examined on day 2, 9, 14, 28 and 35. 【Results】 We found that HCT, MCV, K+, Na+, FHb, hemolytic ratio, 2, 3-DPG and rheological properties in group 0.026 MPa and 0.047 MPa were not significant different compared with that in control (P>0.05). The consumptions of Glu in group 0.026 MPa and 0.047 MPa were significantly higher (P<0.05), and a transient increase in LAC concentration of group 0.026 MPa and 0.047MPa were observed, compared to the control. 【Conclusion】 The conditions of low-pressure and hypoxia have no significant effect on the quality of suspended red blood cells.