Down-regulation of SENP1 Expression Increases Apoptosis of Burkitt Lymphoma Cells

Huang, Bin-Bin,Gao, Qing-Mei,Liang, Wei,Xiu, Bing,Zhang, Wen-Jun,Liang, Ai-Bin Asian Pacific Journal of Cancer Prevention 2012 Asian Pacific journal of cancer prevention Vol.13 No.5

Objective: To investigate the effect of down-regulation of Sentrin/SUMO-specific protease 1 (SENP1) expression on the apoptosis of human Burkitt lymphoma cells (Daudi cells) and potential mechanisms. Methods: Short hairpin RNA (shRNA) targeting SENP1 was designed and synthesized and then cloned into a lentiviral vector. A lentiviral packaging plasmid was used to transfect Daudi cells (sh-SENP1-Daudi group). Daudi cells without transfection (Daudi group) and Daudi cells transfected with blank plasmid (sh-NC-Daudi group) served as control groups. Flow cytometry was performed to screen GFP positive cells and semiquantitative PCR and Western blot assays were employed to detect the inference efficiency. The morphology of cells was observed under a microscope before and after transfection. Fluorescence quantitative PCR and Western blot assays were conducted to measure the mRNA and protein expression of apoptosis related molecules (caspase-3, 8 and 9). After treatment with $COCl_2$ for 24 h, the mRNA and protein expression of hypoxia inducible factor -$1{\alpha}$ (HIF-$1{\alpha}$) was determined. Results: Sequencing showed the expression vectors of shRNA targeting SENP1 to be successfully constructed. Following screening of GFP positive cells by FCM, semiqualitative PCR showed the interference efficiency was $79.2{\pm}0.026%$. At 48 h after transfection, the Daudi cells became shrunken, had irregular edges and presented apoptotic bodies. Western blot assay revealed increase in expression of caspase-3, 8 and 9 with prolongation of transfection (P<0.05). Following hypoxia treatment, mRNA expression of HIF-$1{\alpha}$ remained unchanged in three groups (P>0.05) but the protein expression of HIF-$1{\alpha}$ markedly increased (P<0.05). However, in the sh-SENP1-Daudi group, the protein expression of HIF-$1{\alpha}$ remained unchanged Conclusion: SENP1-shRNA can efficiently inhibit SENP1 expression in Daudi cells. SENP1 inhibition may promote cell apoptosis. These findings suggest that SENP1 may serve as an important target in the gene therapy of Burkitts lymphoma.

Ultrasonic-assisted Solution-Phase Synthesis and Property Studies of Hierarchical Layer-by-Layer Mesoporous CeO2

Pu Su Zhao,Xiu Mei Gao,Feng Xia Zhu,Xin Ming Hu,Lili Zhang 대한화학회 2018 Bulletin of the Korean Chemical Society Vol.39 No.3

Hierarchical layer-by-layer quadrangle CeO2 was prepared through ultrasonic-assisted solution-phase synthesis strategy using cerium oxalate as the precursor. The as-prepared mesoporous CeO2 displayed a surface area of 98.7 m2 g−1 and pore diameters of 2.0–10.0 nm. The high-resolution TEM image revealed that the layer structures of the CeO2 were made of numerous nanocrystal particles with the crystallite size of about 13–15 nm. High energy and cavitation of ultrasonic wave assists cerium oxalate precursor in building the layer-by-layer quadrangle staking. UV–vis absorption spectrum showed that the direct allowed transition bandgap energy for the as-prepared CeO2 was 2.91 eV. Moreover, the CeO2 exhibited good photocatalytic property for degrading Rhodamine B solution under UV radiation.

Histological Changes of Cervical Disc Tissue in Patients with Degenerative Ossification

Yang Xiong,Ying-Li Yang,Yu-Shan Gao,Xiu-Mei Wang,Xing Yu 대한신경외과학회 2022 Journal of Korean neurosurgical society Vol.65 No.2

Objective : To explore the histological feature of the cervical disc degeneration in patients with degenerative ossification (DO) and its potential mechanisms. Methods : A total of 96 surgical segments, from cervical disc degenerative disease patients with surgical treatment, were divided into ossification group (group O, n=46) and non-ossification group (group NO, n=50) based on preoperative radiological exams. Samples of disc tissues and osteophytes were harvested during the decompression operation. The hematoxylin-eosin staining, Masson trichrome staining and Safranin O-fast green staining were used to compare the histological differences between the two groups. And the distribution and content of transforming growth factor (TGF)-β1, p-Smad2 and p-Smad3 between the two groups were compared by a semi-quantitative immunohistochemistry (IHC) method. Results : For all the disc tissues, the content of disc cells and collagen fibers decreased gradually from the outer annulus fibrosus (OAF) to the central nucleus pulposus (NP). Compared with group NO, the number of disc cells in group O increased significantly. But for proteoglycan in the inner annulus fibrosus (IAF) and NP, the content in group O decreased significantly. IHC analysis showed that TGF-β1, p-Smad2, and p-Smad3 were detected in all tissues. For group O, the content of TGF-β1 in the OAF and NP was significantly higher than that in group NO. For p-Smad2 in IAF and p-Smad3 in OAF, the content in group O were significantly higher than group NO. Conclusion : Histologically, cervical disc degeneration in patients with DO is more severe than that without DO. Local higher content of TGF-β1, p-Smad2, and p-Smad3 are involved in the disc degeneration with DO. Further studies with multi-approach analyses are needed to better understand the role of TGF-β/Smads signaling pathway in the disc degeneration with DO.

Cryptotanshinone and wogonin up-regulate eNOS in vascular endothelial cells via ERa and down-regulate iNOS in LPS stimulated vascular smooth muscle cells via ERb

Barnabas Oche,Lu Chen,Ya-ke Ma,Yue Yang,Chun-xiao Li,Xiao Geng,Li-zhen Qiu,Xiu-mei Gao,Hong Wang 대한약학회 2016 Archives of Pharmacal Research Vol.39 No.2

Phytoestrogens were widely used as natural alternatives to estrogen for treating cardiovascular diseases. They have been reported to have cardioprotective and antiinflammatory response, but the mechanisms remain unclear. In this study, we found cryptotanshinone and wogonin exhibited phytoestrogenic property in an estrogen- responsive reporter assay. In EA.hy926 cells, treatment of cryptotanshinone and wogonin led to significant increase in NO production levels, which were inhibited by co-incubation of estrogen receptor (ER)a antagonist methyl-piperidino-pyrazole (MPP). The expression of endothelial NO synthase (eNOS) and ERa were up-regulated with the same treatment, indicating they stimulate NO and eNOS expression via ERa-dependent pathway in endothelial cells. While in lipopolysaccharide activated vascular smooth muscle cell line A7r5, cryptotanshinone and wogonin exerted anti-inflammatory effects by inhibiting NO and inducible NO synthase expression via ERbdependent pathway. The reduction of NO synthesis was not affected by MPP, and was abrogated by ERb antagonist R,R-tetrahydrochrysene. Our findings provide the potential molecular mechanism of cryptotanshinone and wogonin as phytoestrogens for their cardioprotective effects, which exerted regulatory effects on NO synthesis through differential regulation of estrogen receptors. It can be employed as a basis for evaluating the beneficial effects of phytoestrogens in the treatment of patients at risk of cardiovascular disease.

De novo assembly and transcriptome analysis of differentially expressed genes relevant to variegation in hawthorn flowers

Wei Ji,Wei Zhao,Rong‑Chen Liu,Xiao‑Bo Jiao,Kai Han,Zhong‑Yi Yang,Mei‑Ying Gao,Rui Ren,Xiu‑Juan Fan,Ming‑Xia Yang 한국식물생명공학회 2019 Plant biotechnology reports Vol.13 No.6

Flower color variegation has been observed in many plant species. However, pink flowers on the white-blooming hawthorn trees found by our group earlier have never been reported. To better understand the differentially expressed genes (DEGs) in variegated hawthorn flowers, white and pink flowers at different developmental stages (S1 and S2) underwent transcriptome sequencing separately. Approximately 34.28 Gb of high-quality data were obtained and assembled into 100,013 unigenes with an average length of 706.93 bp. These unigenes were further subjected to functional annotation and biochemical pathway analysis, and DEGs of two types of hawthorn flowers at different developmental stages were studied. Based on the enrichment analysis of DEGs, eight anthocyanin-modified enzyme genes or other enzyme genes that indirectly affect anthocyanin synthesis (5AT, 3GGT , and AI, β-Glu, two Aux/IAAs, two PODs), eight structural genes (UFGT, DFR, CHI, two F3Hs, and three PALs), and three transcription factors (one MYB and two bHLHs) were also identified. We randomly selected 15 genes, and the trends in the expression levels of these genes in the organs of white and pink flowers at different developmental stages were verified by quantitative real-time PCR. Mass sequence data obtained by RNA-seq of variegated hawthorn flowers provided basic sequence information and a unique opportunity to uncover the genetic mechanisms under-lying flower color variegation.