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91.
Lin NH Xia P Kovar P Park C Chen Z Zhang H Rosenberg SH Sham HL 《Bioorganic & medicinal chemistry letters》2006,16(2):421-426
Chk1 inhibitors have emerged as a novel class of neoplastic agents for abrogating the G2 DNA damage checkpoint arrest. Analogs of the Chk1 inhibitor, 3-ethylidene-1,3-dihydro-indol-2-one, were synthesized and tested in vitro for their inhibitory activities. The most promising compound identified from this series is analog 28, which possesses potent enzymatic and cellular activities. 相似文献
92.
Shaopeng Chen Junkang Qiu Chuan Chen Chunchun Liu Yuheng Liu Lili An Junying Jia Jie Tang Lijun Wu Haiying Hang 《蛋白质与细胞》2012,3(6):460
Activation-induced cytidine deaminase (AID) is required for the generation of antibody diversity through initiating both somatic hypermutation (SHM) and class switch recombination. A few research groups have successfully used the feature of AID for generating mutant libraries in directed evolution of target proteins in B cells in vitro . B cells, cultured in suspension, are not convenient for transfection and cloning. In this study, we established an AID-based mutant accumulation and sorting system in adherent human cells. Mouse AID gene was first transfected into the human non-small cell lung carcinoma H1299 cells, and a stable cell clone (H1299-AID) was selected. Afterwards, anti-hTNF-αscFv (ATscFv ) was transfected into H1299-AID cells and ATscFv was displayed on the surface of H1299-AID cells. By 4-round amplification/flow cytometric sorting for cells with the highest affinities to hTNF-alpha, two ATscFv mutant gene clones were isolated. Compared with the wild type ATscFv, the two mutants were much more efficient in neutralizing cytotoxicity of hTNF-alpha. The results indicate that directed evolution by somatic hypermutation can be carried out in adherent non-B cells, which makes directed evolution in mammalian cells easier and more efficient. 相似文献
93.
Li Zhang Zhen Li Dongxia Feng Haitao Shen Xiaodi Tian Haiying Li 《Free radical research》2017,51(3):316-328
Oxidative stress is responsible for a poor prognosis of subarachnoid hemorrhage (SAH) patients. Nox2 has been shown to participate in SAH-induced early brain injury (EBI). Nox4 is another major subtype of Nox family widely expressed in central nervous system (CNS). Here, we investigated the role of Nox4 and whether there was a synergistic effect of Nox2 and Nox4 in SAH-induced EBI. Clinical brain biopsies of four patients with traumatic brain injury (TBI) and perihematomal brain tissue from six subjects with SAH were examined. Gp91ds-tat (a specific inhibitor of Nox2), GKT137831 (a specific inhibitor of Nox4), and apocynin (a non-specific Nox inhibitor) were used to test the role of Nox2 and Nox4. The protein levels of Nox2 and Nox4 were elevated in rat neurons and astrocytes at 12?h after SAH, and in cultured brain microvascular endothelial cells at 24?h after exposure to OxyHb. Similarly, there were higher Nox2 and Nox4 protein levels in perihematomal neurons and astrocytes in SAH patients than that in brain tissue from subjects with TBI. In SAH rat model, gp91ds-tat and GKT137831 could reduce SAH-induced neuronal death and degeneration, whereas apocynin did not induce a more intense neuroprotection. Consistently, in in vitro SAH model, siRNA-mediated silencing of Nox2 and Nox4 suppressed the OxyHb-induced neuronal apoptosis, whereas Nox2 and Nox4 co-knockdown also did not show a remarkable overlay effect. In conclusion, Nox4 should contribute to the pathological processes in SAH-induced EBI, and there was not an overlay effect of Nox2 inhibition and Nox4 inhibition on preventing SAH-induced EBI. 相似文献
94.
Lu Beibei Kan Caixia Ke Shanlin Xu Haiying Ni Yuan Wang Changshun Shi Daning 《Plasmonics (Norwell, Mass.)》2017,12(4):1183-1191
Plasmonics - Bimetallic nanoparticles have attracted increasing interest because of their unique optical, electronic, magnetic, and catalytic properties which are different from that of their... 相似文献
95.
麻竹花药培养及再生植株的获得 总被引:2,自引:0,他引:2
以麻竹(Dendrocalamus latiflorus Munro)花药为材料, 于M8+2 mg·L–1 NAA +0.5 mg·L–1 6-BA+15 mg·L–1 PAA+7.5 mg·L–1 STS+500 mg·L–1 CH+100 mg·L–1 proline+100 mg·L–1 glutamin+5.4% maltose+0.8% agar的诱导培养基上成功诱导出胚性愈伤组织, 在此培养基上继代可形成体胚并分化成苗, 初步建立了麻竹花药一步成苗的再生体系。 相似文献
96.
97.
Kou S Pan L van Noort D Meng G Wu X Sun H Xu J Lee I 《Biochemical and biophysical research communications》2011,(2):350-355
Reactive oxygen species (ROS) are an important factor in the development of skin lesions in diabetes. A new antioxidant, hydrogen, can selectively neutralize hydroxyl radicals (OH) and peroxynitrite (ONOO−) in cell-free systems, whereas it seldom reacts with other ROS. Fibroblasts are a key component of skin. In the present study, we investigated the protective effects of hydrogen-rich medium on human skin fibroblasts (HSFs) under oxidative stress. Confocal microscopy was used to assay both the intracellular superoxide anion () concentration and the mitochondrial membrane potential (ΔΨ). Cell viability was determined using the Cell Counting Kit-8 (CCK-8). The concentrations of cellular malonaldehyde (MDA), superoxide dismutase (SOD), glutathione (GSH), 8-hydroxy-2′-deoxyguanosine (8-OHdG) and 3-nitrotyrosine (3-NT) were also measured. The results revealed that both mannitol and high glucose could cause oxidative stress in HSFs. Interestingly, the use of a hydrogen-rich medium significantly reduced the level of intracellular , stabilized the ΔΨ and attenuated production of MDA, 8-OHdG and 3-NT which efficiently enhanced the antioxidative defense system and protected the HSFs from subsequent oxidative stress damage. In other words, hydrogen decreased the excessive generation of intracellular and elevated the cellular antioxidative defense. Based on our results, hydrogen may have applications in the treatment of skin diseases caused by diabetes. 相似文献
98.
The JAK2V617F mutation has been identified in most patients with myeloproliferative neoplasms (MPNs), including polycythemia vera, essential thrombocythemia and primary myelofibrosis. Although JAK2V617F is the predominant allele associated with MPNs, other activating Janus kinase 2 (JAK2) alleles (such as K539L, T875N) also have been identified in distinct MPNs. The basis for the differences in the in vivo effects of different JAK2 alleles remains unclear. We have characterized three different classes of disease-associated JAK2 mutants (JAK2V617F, JAK2K539L and JAK2T875N) and found significant differences in biochemical, signaling and transforming properties among these different classes of JAK2 mutants. 相似文献
99.
Guirong Qiao Renying Zhuo Mingying Liu Jing Jiang Haiying Li Wenmin Qiu Luanyin Pan Sheng lin Xiaoguo Zhang Zongxiu Sun 《Acta Physiologiae Plantarum》2011,33(3):691-696
Soil salinity is a serious problem worldwide. It is necessary to improve the salt tolerance of plants to avoid the progressive
deterioration of saline soil. We showed that the over-expression of AtNHX1 improves salt tolerance in a transgenic poplar (Populus deltoides CL × P. euramericana CL “NL895”) under mannose selection. Four transgenic poplar plants were obtained. Southern blot analysis showed that the
pmi gene had integrated into the genome of the poplar. RT-PCR confirmed that AtNHX1 could be expressed normally in the transgenic plants. When tested for salt tolerance by NaCl stress, we measured a 100%
increase in Na+ content in the three transgenic lines (T18, T50, T98) significantly higher than the 33% increase seen in wild-type plants.
The chlorophyll content of the transgenic plants was not altered significantly, while the chlorophyll content in the control
plants showed a small decrease. MDA content was decreased in the transgenic plants. These results show that the AtNHX1 gene may enhance salt tolerance due to increased vacuolar compartmentalization of sodium ions. 相似文献
100.
Li S Yan Y Zhou Z Yu H Zhan Y Zhang W Chen M Lu W Ping S Lin M 《Molecular biology reports》2011,38(4):2437-2443
The −1 subsite of bacterial fructansucrases (FSs) (levansucrases and inulosucrases) plays an important role in the substrate
recognition, binding and catalysis. Three residues (for example W47, W118 and R193, Zymomonas mobilis levansucrase numbering) at the −1 subsite are completely conserved among FSs. Site-directed mutational analysis showed that
the substitutions of the three strictly conserved amino acid residues, W47N, W47H, W118N, W118H, R193K and R193H, significantly
decreased enzyme activities and synthesis rates of levan, while the size of the synthesized oligosaccharides had been influenced.
These experimental results, combined with 3D structure modeling, lead to our proposal that a single amino acid residue change
in subsite −1 of levansucrase can influence change to the size and polarity of the sucrose binding pocket with a concomitant
change to substrate binding and catalysis, and thus having an overall influence on the enzyme activities and products. 相似文献