1.Construction and identification of nuclear factor erythroid-2-related factor 2 gene RNA interference recombinant plasmid
Journal of Clinical Neurology 2017;30(2):120-123
Objective To construct and identify the shRNA plasmid vector targeting nuclear factor erythroid-2-related factor 2 (Nrf2),and to collect the strongest RNAi effect of Nrf2 shRNA sequence.Methods Nrf2 gene was targeted gene.Three shRNA sequences were designed by software and synthesized by chemical method: shRNA-1,shRNA-2 and shRNA-3.The double strand shRNA oligo was ligated to the vector.The construct was verified by sequencing analysis.BV2 cells were transfected with expressing shRNA plasmid vectors using Lipofectamine 2000.The expression of Nrf2 in the levels of mRNA was detected by real-time PCR,and Western Blot was adopted to abserve the expression of Nrf2 protein.Results Sequencing analysis suggested that the shRNA vectors targeting Nrf2 possessed correct nucleotide sequence and read frame.The result of Real-time PCR and Western Blot showed that the sequence of shRNAi-3 could more effectively knockdown the expression level of Nrf2 than the others.Conclusions The shRNA vectors targeting Nrf2 are successfully constructed and the shRNA can signidicantly inhibit the expression of Nrf2.These findings could provide an experimental basis for further study on Nrf2 signaling pathway in stroke field.
2.Evaluation value of magnetic resonance diffusion tensor imaging for cerebrovascular small vessel disease in patients with systemic lupus erythematosus
Kaidi WU ; Zhanyun REN ; Yiwen SHI ; Wenyun WANG ; Zhenyu QIAN ; Wuzhuang TANG
Chinese Journal of Postgraduates of Medicine 2024;47(2):97-102
Objective:To explore the value of magnetic resonance diffusion tensor imaging (DTI) in evaluating cerebrovascular small vessel disease (CSVD) in patients with systemic lupus erythematosus (SLE).Methods:Eighty-two patients with SLE combined with CSVD treated at Yixing People's Hospital from January to December 2022 were selected. They were divided into acute phase infarction group (16 cases), chronic phase infarction group (26 cases), and chronic ischemic lesion group (40 cases) based on routine MRI examination results. All patients underwent DTI examination to obtain the average diffusion coefficient (DCavg) and anisotropy score (FA) of the affected and contralateral normal white matter areas, and the evaluation value of DTI for CSVD in SLE patients was analyzed.Results:The DCavg value on the affected side of 82 patients was significantly higher than that on the healthy side: (11.10 ± 3.48) 10 -3 mm 2/s vs. (8.18 ± 2.42) 10 -3 mm 2/s, and the FA value on the affected side was significantly lower than that on the healthy side: 0.28 ± 0.05 vs. 0.45 ± 0.08, with a statistical significant differences ( P<0.05). The DCavg values of the acute infarction group, chronic infarction group, and chronic ischemic focus group were (11.88 ± 3.50), (9.69 ± 3.24) and (8.52 ± 2.34) 10 -3 mm 2/s, respectively, with statistical significant differences ( P<0.05). The FA values of the acute infarction group, chronic infarction group, and chronic ischemic focus group were 0.28 ± 0.04, 0.33 ± 0.06 and 0.40 ± 0.07, respectively, with statistical significant differences ( P<0.05). The receiver operating characteristic curve was drawn, and the results showed that the area under the curve (AUC) of the acute phase infarction group and the chronic phase infarction group evaluated by DCavg and FA alone and in combination were 0.757, 0.756, and 0.820, respectively. The AUC of the chronic phase infarction group and the chronic ischemic focus group evaluated by DCavg and FA were 0.772, 0.776, and 0.813, respectively. The AUC value of the combined evaluation was relatively large. Conclusions:DTI has good evaluation value for CSVD in SLE patients and can accurately determine the type of CSVD.