• Asian Pacific Journal of Cancer Prevention
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• v.13 no.5
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• pp.2045-2049
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• 2012

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.

• Journal of the Korean GEO-environmental Society
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• v.2 no.1
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• pp.37-56
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• 2001

As a preliminary study to establish fracture network model in crystalline rocks, detail investigation on fracture characteristics were performed. Five fracture sets were determined on the basis of regional survey of geological structures and fractures on outcrops. Among the fracture sets, S1 set has the highest density and longest trace length of fractures which was identified on surface in the study area. S4 and S5 sets are composed of foliations and foliation parallel shear joints of gneisses, which are very important sets at the aspect of weighting of fracture length. For characterization of subsurface fractures, detail core logging was performed to identify fractures and fracture zones from five boreholes. Acoustic televiewer logging and borehole geophysical loggings produced images, orientations and geophysical properties of fractures which intersect with boreholes. According to the result of the investigations, subsurface fractures can be grouped as three preferred orientations(B1, B2 and B3), which correspond to S1, S2 and S4/S5 of surface fracture sets, respectively. Actually, B1 set is expected to be intensely developed at subsurface. However, it has low frequency of intersection with boreholes due to its parallel or sub-parallel direction to boreholes. According to the inference of conductive fractures, B1 and B3 sets have possibilities of water flow and their intersection lines are also thought to consist of important conduits of groundwater flow. In particular, faults which are parallel to foliations control major groundwater flow in the study area.

• Korean journal of applied entomology
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• v.56 no.4
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• pp.377-385
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• 2017

Estimates of evolutionary sequence divergence and inference of a phylogenetic tree for eight delphacid planthopper species were based on the full-length nucleotide sequence of the internal transcribed spacer 2 (ITS2) region. Size of the ITS2 DNA sequence varied from 550 bp in Sogatella furcifera to 699 bp in Nilaparvata muiri. Nucleotide sequence distance ($d{\pm}S.E.$) was lowest between N. muiri and N. bakeri ($0.001{\pm}0.001$), and highest between Ecdelphax cervina and Stenocranus matsumurai ($0.579{\pm}0.021$). Sequence distance between N. lugens and other planthoppers ranged from $0.056{\pm}0.008$ (N. muiri) to $0.548{\pm}0.021$ (S. matsumurai). In the neighbor-joining phylogenetic tree, all planthoppers were clustered separately into a species group, except N. muiri and N. bakeri. The ITS2 nucleotide sequence of N. lugens was used to design four loop-mediated isothermal amplification (LAMP) primer sets (BPH-38, BPH-38-1, BPH-207, and BPH-92) for N. lugens species-specific detection. After the LAMP reaction of three rice planthoppers, N. lugens, S. furcifera, and Laodelphax striatellus, with the four LAMP primer sets for 60 min at $65^{\circ}C$, LAMP products were observed in the genomic DNA of N. lugens only. In the BPH-92 LAMP primer set, the fluorescence relative to that of the negative control differed according to the amount of DNA (0.1 ng, 10 ng, and 100 ng) and incubation duration (20 min, 30 min, 40 min, and 60 min). At $65^{\circ}C$ incubation, the difference was clearly observed after 40 min with 10 ng and100 ng, but with a 60-min incubation period, the minimum DNA needed was 0.1 ng. However, there was little difference in fluorescence among all DNA amounts tested with 20 or 30 min incubations.