Study on the Molecular Mechanisms of dlk1 Stimulated Lung Cancer Cell Proliferation

Background and objective The imprinted gene dlk1 has been recognized as a cancer related gene since it aberrantly expressed in a series of cancer tissues, but its role in lung cancer is still unknown. The aim of this study is to examine dlk1’s expression in non-small cell lung cancers (NSCLCs) and investigate the molecular mechanism by which dlk1 could accelerate the proliferation of the cells in lung cancer cell lines (H520). Methods The relative expression of dlk1 among 30 NSCLC specimens and their adjacent normal lung tissues were analyzed by RT-PCR. A cell model that stably expressed exogenous dlk1 was established following that the dlk1 gene was cloned into a eukaryotic expression vector and then transfected into the lung cancer cells H520. CCK8 analysis and colony forming assay were employed to investigate the effect of dlk1 on cell proliferation. The expression of CyclinB1 was detected by Western blot. Results dlk1 aberrantly expressed in 36.7% (11/30) of the tumor tissues of NSCLC compared with their adjacent cancer lung tissues. CCK8 analysis showed that overexpression of dlk1 could promote the proliferation of H520 cells (P < 0.05) and the results was further confirmed by colony forming assay. Western blot analysis found that over expression of dlk1 could up-regulate the expression of CyclinB1 (P < 0.05). Conclusion dlk1 aberrantly expressed in NSCLCs. The Overexpression of dlk1 could accelerate the proliferation of lung cancer cells H520 in vitro, probably through up-regulating the expression of cell cycle protein CyclinB1

Reconciling results of 2019 and 2020 stellar occultations on Pluto's atmosphere. New constraints from both the 5 September 2019 event and consistency analysis

A stellar occultation by Pluto on 5 September 2019 yielded positive detections at two separate stations. Using an approach consistent with comparable studies, we derived a surface pressure of $11.478 \pm 0.55~\mathrm{\mu bar}$ for Pluto's atmosphere from the observations of this event. In addition, to avoid potential method inconsistancies highlighted by Sicardy et al. when comparing with historical pressure measurements, we reanalyzed the data by 15 August 2018 and 17 July 2019 events, respectively. All the new measurements provide a bridge between the two different perspectives on the pressure variation since 2015: a rapid pressure drop from previous studies of the 15 August 2018 and 17 July 2019 events and a plateau phase from that of the 6 June 2020 event. The pressure measurement from the 5 September 2019 event aligns with those from 2016, 2018, and 2020, supporting the latter perspective. While the measurements from the 4 June 2011 and 17 July 2019 events suggest probable V-shaped pressure variations unaccounted for by the volatile transport model (VTM) from Meza et al., the VTM remains applicable on average. And, the validity of the V-shaped variations is debatable due to the stellar faintness of the 4 June 2011 event and the grazing single-chord geometry of the 17 July 2019 event. To reveal and understand all significant pressure variations of Pluto's atmosphere, it is essential to provide constraints on both short-term and long-term evolutions of the interacting atmosphere and surface by continuous pressure monitoring through occultation observations, whenever possible, complemented by frequent spectroscopy and photometry of the surface.Comment: Accepted for publication in Astronomy & Astrophysics. 10 pages, 6 figure

Effect of Nano Zeolite on the Transformation of Cadmium Speciation and Its Uptake by Tobacco in Cadmium-contaminated Soil

The study was aimed at exploring the effects of applying ordinary and nano zeolite on the soil pH, soil available cadmium (Cd) content, soil Cd speciation and Cd uptakes by tobacco using pot experiment with simulated Cd contaminated soil indoors. The results showed that soil pH increased and available Cd content reduced with the amount of ordinary and nano zeolite increasing. Compared with the control, the application of ordinary and nano zeolite increased soil pH at 0.47 - 1.05 and 0.73 - 1.57, respectively, and reduced the available Cd contents at 19.3% - 32.7% and 23.2% - 40.5%, respectively. In addition, soil pH had significantly negative correlation with available Cd content in each treatment (p<0.05). Nano zeolite could more effectively reduce Cd contents of all parts of tobacco than ordinary zeolite with the same amount treatments, and Cd content in all parts of tobacco plants was positively correlated with soil available Cd content. The content of exchangeable Cd (EXE) in soil decreased to some extent with different zeolite treatments, application of nano zeolite was better than that of application of ordinary zeolite in reducing Cd bioavailability and transferability. Overall, application of nano zeolite has an advantage over ordinary zeolite in reducing available Cd content in soil and Cd content in tobacco

Mutation of <i>Leaf Senescence 1</i> Encoding a C2H2 Zinc Finger Protein Induces ROS Accumulation and Accelerates Leaf Senescence in Rice

Premature senescence of leaves causes a reduced yield and quality of rice by affecting plant growth and development. The regulatory mechanisms underlying early leaf senescence are still unclear. The Leaf senescence 1 (LS1) gene encodes a C2H2-type zinc finger protein that is localized to both the nucleus and cytoplasm. In this study, we constructed a rice mutant named leaf senescence 1 (ls1) with a premature leaf senescence phenotype using CRISPR/Cas9-mediated editing of the LS1 gene. The ls1 mutants exhibited premature leaf senescence and reduced chlorophyll content. The expression levels of LS1 were higher in mature or senescent leaves than that in young leaves. The contents of reactive oxygen species (ROS), malondialdehyde (MDA), and superoxide dismutase (SOD) were significantly increased and catalase (CAT) activity was remarkably reduced in the ls1 plants. Furthermore, a faster decrease in pigment content was detected in mutants than that in WT upon induction of complete darkness. TUNEL and staining experiments indicated severe DNA degradation and programmed cell death in the ls1 mutants, which suggested that excessive ROS may lead to leaf senescence and cell death in ls1 plants. Additionally, an RT-qPCR analysis revealed that most senescence-associated and ROS-scavenging genes were upregulated in the ls1 mutants compared with the WT. Collectively, our findings revealed that LS1 might regulate leaf development and function, and that disruption of LS1 function promotes ROS accumulation and accelerates leaf senescence and cell death in rice

Novel Salinity-Tolerant Third-Generation Hybrid Rice Developed via CRISPR/Cas9-Mediated Gene Editing

Climate change has caused high salinity in many fields, particularly in the mud flats in coastal regions. The resulting salinity has become one of the most significant abiotic stresses affecting the world’s rice crop productivity. Developing elite cultivars with novel salinity-tolerance traits is regarded as the most cost-effective and environmentally friendly approach for utilizing saline-alkali land. To develop a highly efficient green strategy and create novel rice germplasms for salt-tolerant rice breeding, this study aimed to improve rice salinity tolerance by combining targeted CRISPR/Cas9-mediated editing of the OsRR22 gene with heterosis utilization. The novel alleles of the genic male-sterility (GMS) and elite restorer line (733Srr22-T1447-1 and HZrr22-T1349-3) produced 110 and 1 bp deletions at the third exon of OsRR22 and conferred a high level of salinity tolerance. Homozygous transgene-free progeny were identified via segregation in the T2 generation, with osrr22 showing similar agronomic performance to wild-type (733S and HZ). Furthermore, these two osrr22 lines were used to develop a new promising third-generation hybrid rice line with novel salinity tolerance. Overall, the results demonstrate that combining CRISPR/Cas9 targeted gene editing with the “third-generation hybrid rice system” approach allows for the efficient development of novel hybrid rice varieties that exhibit a high level of salinity tolerance, thereby ensuring improved cultivar stability and enhanced rice productivity