Screening for the optimal siRNA targeting a novel gene (HA117) and construction and evaluation of a derivative recombinant adenovirus

We found a novel gene named as HA117 in our previous research. At this study, we screened for an optimal siRNA targeting the novel gene HA117 using the pSOS-HUS method, verified the results of pSOS-HUS siRNA screening for optimal affinity for the target gene, and constructed and evaluated a recombinant adenovirus carrying the DNA template for transcription of the optimal HA117 siRNA. The pSOS-HUS vector method was successfully utilized as a rapid and effective screen for an optimal siRNA for a target gene. Among five pairs of DNA templates, siRNA transcribed from HAi5 gave the strongest interference with the novel gene HA117; a HAi5-carrying recombinant adenovirus (Ad-HAi5) was successfully constructed and evaluated, laying a foundation for the further study of HA117 gene function with RNAi technology

Glycogen synthase kinase-3β inhibition induces nuclear factor-κB-mediated apoptosis in pediatric acute lymphocyte leukemia cells

<p>Abstract</p> <p>Background</p> <p>Molecular therapies that target genetic abnormalities in leukemic cells and their affected signaling pathways have been emerging in pediatric acute lymphoblastic leukemia (ALL). Glycogen synthase kinase-3β (GSK-3β) has recently been found to positively regulate the activity of nuclear factor-κB (NF-κB). Here, we investigated the relationship between GSK-3β inhibition and NF-κB in apoptosis of pediatric primary leukemia cells obtained from 39 newly diagnosed ALL children in China.</p> <p>Methods</p> <p>Bone marrow mononuclear cells (BMMC) were isolated by density gradient centrifugation from the heparinized aspirates of children with ALL. We used immunofluorescence staining to detect nuclear GSK-3β in these cells. After treatment with chemically distinct GSK-3β inhibitors in vitro, NF-κB transcriptional activity was identified by means of western blotting and electrophoretic mobility shift assay (EMSA). NF-κB-mediated apoptosis was detected by Annexin V-PE/7-AAD double-staining flow cytometry. The expression level of the <it>survivin </it>gene was detected by reverse-transcriptase polymerase chain reaction (RT-PCR).</p> <p>Results</p> <p>GSK-3β significantly accumulates in the nuclei of ALL cells than in the nuclei of control cells. Cell death induced by GSK-3β inhibition in ALL cells was mediated by a downregulation of NF-κB p65 transcriptional activity. GSK-3β inhibition significantly decreased the expression of the NF-κB target gene <it>survivin</it>.</p> <p>Conclusions</p> <p>These results indicate that inhibition of GSK-3β downregulates the NF-κB activation pathway, leading to suppression of the expression of an NF-κB-regulated gene and promotion of apoptosis in ALL cells in vitro. Furthermore, our findings suggest that GSK-3β or NF-κB is a potential therapeutic target in the treatment of pediatric ALL.</p

Simulation of CSSTs astrometric capability

The China Space Station Telescope (CSST) will enter a low Earth orbit around 2024 and operate for 10 years, with seven of those years devoted to surveying the area of the median-to-high Galactic latitude and median-to-high Ecliptic latitude of the sky. To maximize the scientific output of CSST, it is important to optimize the survey schedule. We aim to evaluate the astrometric capability of CSST for a given survey schedule and to provide independent suggestions for the optimization of the survey strategy. For this purpose, we first construct the astrometric model and then conduct simulated observations based on the given survey schedule. The astrometric solution is obtained by analyzing the simulated observation data. And then we evaluate the astrometric capability of CSST by analyzing the properties of the astrometric solution. We find that the accuracy of parallax and proper motion of CSST is better than 1 mas( yr1) for the sources of 18-22 mag in g band, and about 1-10 mas( yr1) for the sources of 22-26 mag in g band, respectively. The results from real survey could be worse since the assumptions are optimistic and simple. We find that optimizing the survey schedule can improve the astrometric accuracy of CSST. In the future, we will improve the astrometric capability of CSST by continuously iterating and optimizing the survey schedule.Comment: 17 pages, 10 figure

Klotho-derived peptide KP1 ameliorates SARS-CoV-2-associated acute kidney injury

Introduction: The severe cases of COVID-19, a disease caused by severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2), often present with acute kidney injury (AKI). Although old age and preexisting medical conditions have been identified as principal risk factors for COVID-19-associated AKI, the molecular basis behind such a connection remains unknown. In this study, we investigated the pathogenic role of Klotho deficiency in COVID-19-associated AKI and explored the therapeutic potential of Klotho-derived peptide 1 (KP1).Methods: We assessed the susceptibility of Klotho deficient Kl/Kl mice to developing AKI after expression of SARS-CoV-2 N protein. The role of KP1 in ameliorating tubular injury was investigated by using cultured proximal tubular cells (HK-2) in vitro and mouse model of ischemia-reperfusion injury (IRI) in vivo.Results: Renal Klotho expression was markedly downregulated in various chronic kidney disease (CKD) models and in aged mice. Compared to wild-type counterparts, mutant KL/KL mice were susceptible to overexpression of SARS-CoV-2 N protein and developed kidney lesions resembling AKI. In vitro, expression of N protein alone induced HK-2 cells to express markers of tubular injury, cellular senescence, apoptosis and epithelial-mesenchymal transition, whereas both KP1 and Klotho abolished these lesions. Furthermore, KP1 mitigated kidney dysfunction, alleviated tubular injury and inhibited apoptosis in AKI model induced by IRI and N protein.Conclusion: These findings suggest that Klotho deficiency is a key determinant of developing COVID-19-associated AKI. As such, KP1, a small peptide recapitulating Klotho function, could be an effective therapeutic for alleviating AKI in COVID-19 patients

CSST forecast: impact from non-Gaussian covariances and requirements on systematics-control

The precise estimation of the statistical errors and accurate removal of the systematical errors are the two major challenges for the stage IV cosmic shear surveys. We explore their impact for the China Space-Station Telescope (CSST) with survey area $\sim17,500\deg^2$ up to redshift $\sim4$. We consider statistical error contributed from Gaussian covariance, connected non-Gaussian covariance and super-sample covariance. We find the super-sample covariance can largely reduce the signal-to-noise of the two-point statistics for CSST, leading to a $\sim1/3$ loss in the figure-of-merit for the matter clustering properties ($\sigma_8-\Omega_m$ plane) and $1/6$ in the dark energy equation-of-state ($w_0-w_a$ plane). We further put requirements of systematics-mitigation on: intrinsic alignment of galaxies, baryonic feedback, shear multiplicative bias, and bias in the redshift distribution, for an unbiased cosmology. The $10^{-2}$ to $10^{-3}$ level requirements emphasize strong needs in related studies, to support future model selections and the associated priors for the nuisance parameters.Comment: submitted to MNRA

Effects of fertilizer application schemes and soil environmental factors on nitrous oxide emission fluxes in a rice-wheat cropping system, east China

Nitrous oxide (N2O) is a potent greenhouse gas (GHG) with agricultural soils representing its largest anthropogenic source. However, the mechanisms involved in the N2O emission and factors affecting N2O emission fluxes in response to various nitrogenous fertilizer applications remain uncertain. We conducted a four-year (2012–2015) field experiment to assess how fertilization scheme impacts N2O emissions from a rice-wheat cropping system in eastern China. The fertilizer treatments included Control (CK), Conventional fertilizer (CF), CF with shallow-irrigation (CF+SI), CF with deep-irrigation system (CF+DI), Optimized fertilizer (OF), OF with Urease inhibitor (OF+UI), OF with conservation tillage (OF+CT) and Slow-release fertilizer (SRF). N2O emissions were measured by a closed static chamber method. N2O emission fluxes ranged from 0.61 μg m-2 h-1 to 1707 μg m-2 h-1, indicating a significant impact of nitrogen fertilizer and cropping type on N2O emissions. The highest crop yields for wheat (3515–3667 kg ha-1) and rice (8633–8990 kg ha-1) were observed under the SRF and OF+UI treatments with significant reduction in N2O emissions by 16.94–21.20% and 5.55–7.93%, respectively. Our findings suggest that the SRF and OF+UI treatments can be effective in achieving maximum crop yield and lowering N2O emissions for the rice-wheat cropping system in eastern China