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101.
Chenyang Han Yi Yang Qiaobing Guan Xiaoling Zhang Heping Shen Yongjia Sheng Jin Wang Xiaohong Zhou Wenyan Li Li Guo Qingcai Jiao 《Journal of cellular and molecular medicine》2020,24(14):8078-8090
The present study was designed to investigate the role of β‐amyloid (Aβ1‐42) in inducing neuronal pyroptosis and its mechanism. Mice cortical neurons (MCNs) were used in this study, LPS + Nigericin was used to induce pyroptosis in MCNs (positive control group), and Aβ1‐42 was used to interfere with MCNs. In addition, propidium iodide (PI) staining was used to examine cell permeability, lactate dehydrogenase (LDH) release assay was employed to detect cytotoxicity, immunofluorescence (IF) staining was used to investigate the expression level of the key protein GSDMD, Western blot was performed to detect the expression levels of key proteins, and enzyme‐linked immunosorbent assay (ELISA) was utilized to determine the expression levels of inflammatory factors in culture medium, including IL‐1β, IL‐18 and TNF‐α. Small interfering RNA (siRNA) was used to silence the mRNA expression of caspase‐1 and GSDMD, and Aβ1‐42 was used to induce pyroptosis, followed by investigation of the role of caspase‐1‐mediated GSDMD cleavage in pyroptosis. In addition, necrosulfonamide (NSA), an inhibitor of GSDMD oligomerization, was used for pre‐treatment, and Aβ1‐42 was subsequently used to observe the pyroptosis in MCNs. Finally, AAV9‐siRNA‐caspase‐1 was injected into the tail vein of APP/PS1 double transgenic mice (Alzheimer's disease mice) for caspase‐1 mRNA inhibition, followed by observation of behavioural changes in mice and measurement of the expression of inflammatory factors and pyroptosis‐related protein. As results, Aβ1‐42 could induce pyroptosis in MCNs, increase cell permeability and enhance LDH release, which were similar to the LPS + Nigericin‐induced pyroptosis. Meanwhile, the expression levels of cellular GSDMD and p30‐GSDMD were up‐regulated, the levels of NLRP3 inflammasome and GSDMD‐cleaved protein caspase‐1 were up‐regulated, and the levels of inflammatory factors in the medium were also up‐regulated. siRNA intervention in caspase‐1 or GSDMD inhibited Aβ1‐42‐induced pyroptosis, and NSA pre‐treatment also caused the similar inhibitory effects. The behavioural ability of Alzheimer's disease (AD) mice was relieved after the injection of AAV9‐siRNA‐caspase‐1, and the expression of pyroptosis‐related protein in the cortex and hippocampus was down‐regulated. In conclusion, Aβ1‐42 could induce pyroptosis by GSDMD protein, and NLRP3‐caspase‐1 signalling was an important signal to mediate GSDMD cleavage, which plays an important role in Aβ1‐42‐induced pyroptosis in neurons. Therefore, GSDMD is expected to be a novel therapeutic target for AD. 相似文献
102.
广藿香根与根茎挥发油成分研究 总被引:7,自引:0,他引:7
应用气相色谱-质谱联用技术对广藿香根和根茎中挥发油成分进行了分析,从根油中分离出50个色谱峰,鉴定了47个化合物,占总量的95.02%,主要含广藿香酮(81.71%)、d-苦橙油醇(2.88%)、十六烷酸(2.21%)、广藿香醇(1.98%)和3,7,11-三甲基-2,6,10-十二烷三烯-2-醇乙酸酯(1.09%)、广藿香醇(1.98%)和3,7,11-三甲基-2,6,10-十二烷三烯-1-醇乙 相似文献
103.
CO2 biofixation and fatty acid composition of Scenedesmus obliquus and Chlorella pyrenoidosa in response to different CO2 levels 总被引:1,自引:0,他引:1
In this study, Scenedesmus obliquus SJTU-3 and Chlorella pyrenoidosa SJTU-2 were cultivated with 0.03%, 5%, 10%, 20%, 30%, 50% CO(2). The two microalgae could grow at 50% CO(2) (>0.69 g L(-1)) and grew well (>1.22 g L(-1)) under CO(2) concentrations ranging from 5% to 20%. Both of the two examined microalgae showed best growth potential at 10% CO(2). The maximum biomass concentration and CO(2) biofixation rate were 1.84 g L(-1) and 0.288 g L(-1) d(-1) for S. obliquus SJTU-3 and 1.55 g L(-1) and 0.260 g L(-1) d(-1) for C. pyrenoidosa SJTU-2, respectively. The main fatty acid compositions of the two examined microalgae were fatty acids with C(16)-C(18) (>94%) under different CO(2) levels. High CO(2) levels (30-50%) were favorable for the accumulation of total lipids and polyunsaturated fatty acids. The present results suggested that the two microalgae be appropriate for mitigating CO(2) in the flue gases and biodiesel production. 相似文献
104.
Establishing a dual knock-out cell line by lentivirus based combined CRISPR/Cas9 and Loxp/Cre system
Ya Li Weifeng Zhang Junli Zhao Sai Li Linlin Shan Jiuling Zhu Yan Li He Zhu Qinwen Mao Haibin Xia 《Cytotechnology》2018,70(6):1595-1605
The clustered regulatory interspersed short palindromic repeat (CRISPR)/CRISPR-associated protein (Cas) system has been widely used for gene knock-out. Lentiviral vectors have been commonly used as a delivery method for this system, however, prolonged Cas9/sgRNA expression due to lentiviral integration can lead to accumulating off-target mutations. To solve this issue in engineering a gene knock-out cell line, this study established a novel system, which was composed of two lentiviral vectors. One lentiviral vector carried simultaneously sgRNAs and CRISPR/Cas9 expression cassettes targeting single or multiple gene(s); the other lentiviral vector carried Cre that could remove excess sgRNAs and Cas9 expression cassettes in the genome after gene targeting was achieved. To prove the principle, two candidate genes, extracellular matrix protein 1 (ECM1) and progranulin (PGRN), both highly expressed in MDA-MB-231 cells, were selected for testing the novel system. A dual knock-out of ECM1 and PGRN was successfully achieved in MDA-MB-231 cell line, with the sgRNAs and Cas9 expression cassettes being removed by Cre. This system should have great potential in applications for multiple genes knock-out in vitro. 相似文献
105.
Hasegawa K Tamari M Shao C Shimizu M Takahashi N Mao XQ Yamasaki A Kamada F Doi S Fujiwara H Miyatake A Fujita K Tamura G Matsubara Y Shirakawa T Suzuki Y 《Human genetics》2004,115(4):295-301
Bronchial asthma (BA) is a common chronic inflammatory disease characterized by hyperresponsive airways, excess mucus production, eosinophil activation, and the production of IgE. The complement system plays an immunoregulatory role at the interface of innate and acquired immunities. Recent studies have provided evidence that C3, C3a receptor, and C5 are linked to airway hyperresponsiveness. To determine whether genetic variations in the genes of the complement system affect susceptibility to BA, we screened single nucleotide polymorphisms (SNPs) in C3, C5, the C3a receptor gene (C3AR1), and the C5a receptor gene (C5R1) and performed association studies in the Japanese population. The results of this SNP case-control study suggested an association between 4896C/T in the C3 gene and atopic childhood BA (P=0.0078) as well as adult BA (P=0.010). When patient data were stratified according to elevated total IgE levels, 4896C/T was more closely associated with adult BA (P=0.0016). A patient-only association study suggested that severity of childhood BA was associated with 1526G/A of the C3AR1 gene (P=0.0057). We identified a high-risk haplotype of the C3 gene for childhood (P=0.0021) and adult BA (P=0.0058) and a low-risk haplotype for adult BA (P=0.00011). We also identified a haplotype of the C5 gene that was protective against childhood BA (P=1.4×10–6) and adult BA (P=0.00063). These results suggest that the C3 and C5 pathways of the complement system play important roles in the pathogenesis of BA and that polymorphisms of these genes affect susceptibility to BA. 相似文献
106.
Longxi Mao Shilian Le Xin Jin Guoliang Liu Jun Chen Junwen Hu 《Experimental cell research》2019,374(2):274-281
COP9 signalosome subunit 5 (CSN5) has been involved in the progression of diverse human cancers. MMP2 plays an important role in the metastasis of cancer cells. However, the roles and relationship of in pancreatic cancer (PC) is still unknown. Here, our data shown that both CSN5 and MMP2 were significantly upregulated in PC compared with the corresponding adjacent tissues, where a positive correlation in their expression and associated malignant characteristics were found. Further, silencing of CSN5 expression markedly inhibited PC invasion and metastasis in vitro and in vivo, accompanied by decreased MMP2 expression. Moreover, the anti-metastasis role of CSN5 silence was reversed by MMP2 overexpression, whereas knockdown of MMP2 decreased PC metastasis driven by upregulation of CSN5. Further investigation revealed that CSN5 regulated MMP2 expression via activation of FOXM1 in PC cells. Mechanistically, CSN5 directly bound FOXM1 and decreased its ubiquitination to enhance the protein stability of FOXM1. Taken together, the results indicate that CSN5 can contribute to PC invasion and metastasis through activation of FOXM1/MMP2 axis. 相似文献
107.
Chen Y Ip SP Ko KM Poon TC Ng EW Lai PB Mao QQ Xian YF Che CT 《Journal of proteome research》2011,10(1):299-304
To identify key proteins involved in the hepatoprotection afforded by schisandrin B (Sch B), we used a proteomic approach to screen proteins that were specifically regulated by Sch B in mouse livers and to investigate the role of the proteins in hepatoprotection. Thirteen proteins were specifically activated or suppressed by Sch B treatment. Among the 13 proteins, Raf kinase inhibitor protein (RKIP) was postulated to be the key regulator involved in the development of hepatotoxin-induced cellular damage. The results indicated that the downregulation of RKIP by antisense RKIP vector transfection led to the activation of the Raf-1/MEK/ERK signaling pathway, as evidenced by increases in the level of MEK/ERK phosphorylation and the level of nuclear factor erythroid 2-related factor 2 in the nucleus. The signaling effect produced by RKIP downregulation resembled that triggered by Sch B, wherein both treatments resulted in a decrease in the extent of carbon tetrachloride-induced apoptotic cell death in AML12 hepatocytes. Overexpression of RKIP by the sense RKIP transfection vector or the inhibition of MEK kinase by PD98059 was able to abrogate the cytoprotective effect of Sch B in the hepatocytes. The results indicate that Sch B triggers the Raf/MEK/ERK signaling pathway, presumably by downregulating RKIP, thereby protecting against carbon tetrachloride-induced cytotoxicity. 相似文献
108.
109.
子宫颈糜烂病毒病因的探讨 总被引:17,自引:0,他引:17
491份宫颈拭子病毒分离结果表明:糜烂宫颈单纯疱疹病毒(HSV)分离阳性率(30.8%)是正常宫颈(2.6%)的11.8倍,用人干扰素治疗一个疗程后,病毒分离率下降至疗前的1/4.36例糜烂宫颈活体组织DNA分子杂交表明,乳头瘤病毒16型(HPV-16)阳性者占52.8和,HPV-18占17.9%,HPV-6B占28.1%,HPV-11占7.7%,251例宫颈糜烂患者经人(?)D型基因工程干扰素双盲对比治疗后,总有效率达93.8%,显效率达60%,分析临床疗效与HSV分离率的变化表明,临床有效病例中有35%(49/140)在治疗后病毒阴转,有57%在疗前疗后均未分离出HSV,有5%在疗前疗后保持阳性不变,有2.9%疗前阴性,疗后阳性,上述结果表明,HSV和HPV与慢性宫颈炎有一定关系。 相似文献
110.
A novel water-soluble dextran was synthesized from maltodextrin by cell-free extract of Gluconobacter oxydans DSM 2003. The dextran was purified by size exclusion chromatography, and the structure was determined by Fourier transform infrared spectroscopy, nuclear magnetic resonance, and gas chromatography-mass spectrometer. Based on the spectral data, we found that the dextran contained only D-glucose residues. The ratio of nonreducing end glucopyranosyl (Glcp) to 6-linked Glcp to 4,6-linked Glcp was estimated to be 8.62:78.79:12.59 by methylation analysis. This result indicated the existence of a small proportion of α(1,4) branches in α(1,6) glucosyl linear chains. Here, we reported the first time a novel dextran was synthesized by G. oxydans DSM 2003. 相似文献