An effective method for establishing a regeneration and genetic transformation system for Actinidia arguta

The all-red A. arguta (Actinidia arguta) is an anthocyanin-rich and excellent hardy fruit. Many studies have focused on the green-fleshed A. arguta, and fewer studies have been conducted on the all-red A. arguta. Here we reported a regeneration and Agrobacterium-mediated transformation protocol by using leaves of all-red A. arguta as explants. Aseptic seedling leaves of A. arguta were used as callus-inducing materials. MS medium supplemented with 0.3 mg·L-1 2,4-D and 1.0 mg·L-1 BA was the optimal medium for callus induction of leaves, and medium supplemented with 3 mg·L-1 tZ and 0.5 mg·L-1 IAA was optimal for adventitious shoot regeneration. The best proliferation medium for adventitious buds was MS + 1.0 mg·L-1 BA + 0.3 mg·L-1 NAA. The best rooting medium was 1/2MS + 0.7 mg·L-1 IBA with a 100% rooting rate. For the red flesh hardy kiwi variety ‘Purpurna Saduwa’ (A. arguta var. purpurea), leaves are receptors for Agrobacterium (EHA105)-mediated transformation. The orthogonal experiment was used for the optimization of each genetic transformation parameter and the genetic transformation of the leaves was 21% under optimal conditions. Our study provides technical parameters for applying genetic resources and molecular breeding of kiwifruit with red flesh

Impact of Family Functioning on Adolescent Materialism and Egocentrism in Mainland China: Positive Youth Development Attributes as a Mediator

Although adolescent materialism and egocentrism are growing problems in Chinese societies, there are very few studies investigating their predictors and related mediators. Longitudinal studies in this area are also sparse. Based on a short-term longitudinal study (n = 4981), we assessed the impact of family functioning on materialism and egocentrism of Chinese adolescents, with positive youth development attributes as a hypothesized mediating factor. Results showed that family functioning positively predicted positive youth development attributes but negatively predicted adolescent materialism and egocentrism; positive youth development attributes also negatively predicted adolescent materialism and egocentrism. Mediational analyses showed that positive youth development attributes mediated the impact of family functioning on adolescent materialism and egocentrism. The theoretical and methodological advances of the study are discussed

Analysis of post-traumatic growth status and its influencing factors in patients with facial palsy

Abstract Background Facial nerve paralysis in patients occurs in varying degrees of self-image disorders, both physically and mentally, resulting in low self-esteem, anxiety, depression, and even suicide; however, there were few researches on psychological problems in facial palsy patients at home and abroad. This study’s objective was to investigate post-traumatic growth (PTG) in facial nerve palsy patients and analyze its influencing factors. Methods Using the convenience sampling method, a total of 47 patients with facial nerve paralysis were enrolled in the current study between June 1, 2016, and May 31, 2017. Post-traumatic growth rating scale was utilized to investigate the post-traumatic growth of these patients, and factors influencing patients’ post-traumatic growth were analyzed through collecting the general sociological information, disease-related information, simple coping style questionnaire, and social support rating scale. Results The total score of post-traumatic growth in patients with facial nerve paralysis was mean (M) = 63.1, standard deviation (SD) = 19.14. The ranking of five dimensional scores from high to low was as follows: new possibilities, personal strength enhancement, appreciation of life, mental changes, and improvement of relationships with others. Multiple linear regression analysis showed that six variables, namely, the personality type, duration with facial nerve paralysis, and four coping styles, consisting of three types of positive coping styles and one negative coping style, could explain 71.6% of the total post-traumatic growth score. Conclusions Post-traumatic growth in facial nerve palsy patients is moderate. The personality type of patients, the disease duration, and the coping style are the primary influencing factors. Therefore, clinical staffs should perform personalized nursing protocol and psychological intervention for facial nerve palsy patients to reduce their negative mood, improve their compliance with treatment, and help them recover more rapidly

Generation of Large-Scale Moderate-Resolution Forest Height Mosaic With Spaceborne Repeat-Pass SAR Interferometry and Lidar

This paper provides an overview of the scattering model, inversion approach, and validation of the application results for creating large-scale moderate-resolution (hectare-level) mosaics of forest height through using spaceborne repeat-pass SAR interferometry and lidar. By incorporating several improvements to the forest height inversion and mosaicking approach, the height estimation accuracy along with the robustness of this approach have been considerably enhanced from its originally reported accuracy of RMSE of 3–4 m at a 20-hectare aggregated pixel size to RMSE of 3–4 m on the order of 3–6 hectares. Furthermore, practical data processing schemes are provided in detail. Extensive validation results are demonstrated which include: 1) a forest height mosaic (total area of 11.6 million hectares) is generated for the U.S. states of Maine and New Hampshire using Japanese Aerospace Exploration Agency’s (JAXA) ALOS-1 InSAR correlation data and a small airborne lidar strip (44 000 hectares); 2) the mosaic height estimates are further compared with the available airborne lidar data and field measurements over both flat and mountainous areas; and 3) feasibility of using modern repeat-pass InSAR satellites with short repeat interval is also examined by using JAXA’s ALOS-2 data. This simple and efficient approach is a potential observational prototype with much smaller error budget for the future spaceborne repeat-pass L-band InSAR systems with small spatial baseline and moderate/large temporal baseline (such as NISAR) in combination with lidar (such as GEDI) on the application of large-scale forest height/biomass mapping. It also serves as a complementary tool to the spaceborne single-pass InSAR systems using InSAR/PolInSAR methods when full-pol data are not available and/or when the underlying topography slope causes problems for these approaches

Generation of Large-Scale Moderate-Resolution Forest Height Mosaic With Spaceborne Repeat-Pass SAR Interferometry and Lidar

This paper provides an overview of the scattering model, inversion approach, and validation of the application results for creating large-scale moderate-resolution (hectare-level) mosaics of forest height through using spaceborne repeat-pass SAR interferometry and lidar. By incorporating several improvements to the forest height inversion and mosaicking approach, the height estimation accuracy along with the robustness of this approach have been considerably enhanced from its originally reported accuracy of RMSE of 3–4 m at a 20-hectare aggregated pixel size to RMSE of 3–4 m on the order of 3–6 hectares. Furthermore, practical data processing schemes are provided in detail. Extensive validation results are demonstrated which include: 1) a forest height mosaic (total area of 11.6 million hectares) is generated for the U.S. states of Maine and New Hampshire using Japanese Aerospace Exploration Agency’s (JAXA) ALOS-1 InSAR correlation data and a small airborne lidar strip (44 000 hectares); 2) the mosaic height estimates are further compared with the available airborne lidar data and field measurements over both flat and mountainous areas; and 3) feasibility of using modern repeat-pass InSAR satellites with short repeat interval is also examined by using JAXA’s ALOS-2 data. This simple and efficient approach is a potential observational prototype with much smaller error budget for the future spaceborne repeat-pass L-band InSAR systems with small spatial baseline and moderate/large temporal baseline (such as NISAR) in combination with lidar (such as GEDI) on the application of large-scale forest height/biomass mapping. It also serves as a complementary tool to the spaceborne single-pass InSAR systems using InSAR/PolInSAR methods when full-pol data are not available and/or when the underlying topography slope causes problems for these approaches

Stable knockout of Mcart-1 down-regulates the proliferation and oxidative phosphorylation of RAW264.7 macrophages

Objective To construct a RAW264.7 macrophage cell strain with stable knockout of Mcart-1 by using CRISPR/Cas9 gene editing technique, and to detect its biological function. Methods Cas9 and sgRNA lentivirus were used to infect RAW264.7 macrophages in two steps. Positive cells were screened with puromycin and hygromycin, and single cells were plated by flow cytometry to obtain monoclonal cells; Expression of Cas9 and Mcart-1 was detected by qPCR and Western blot, and the mutation site was confirmed by sequence analysis. Cells number counting and carboxyfluorescein diacetate succinimdyl ester (CFSE) staining were used to detect cell proliferation; ATP detection kit was used to measure total ATP content in cells; Seahorse bioenergy analyzer was used to detect cell oxygen consumption rate(OCR) and glycolysis rate. Results Successfully constructed RAW264.7 cell strain with Mcart-1 stably knocked out, denoted as Mcart-1-/--RAW264.7; A frameshift mutation occurred in Mcart-1 gene in this cell strain; Compared with the wild-type RAW264.7 cell line (RAW264.7), the OCR increased and the extracellular acidification rate(ECAR)of Mcart-1-/--RAW264.7 cells decreased; The total ATP content in Mcart-1 knocked out cells decreased(P＜0.05),mitochondrial ATP production rate decreased (P＜0.01), while glycolytic ATP production rate increased (P＜0.01); The proliferation activity of RAW264.7 cells decreased after Mcart-1 was knocked out(P＜0.001). Conclusions The proliferation activity and oxidative phosphorylation level of RAW264.7 cells were both down-regulated after the Mcart-1 was stably knocked out. This cell strain is an important tool for exploring the function of cells in the tumor microenvironment

Culture of glioblastoma U87 stem-like cells and identification of its metabolic phenotype and tumorigenic ability

Objective To cultivate glioblastoma U87 stem-like cells (SLCs) and to detect the level of stemness biomarkers, mitochondrial respiratory capacity and the capacity of in vivo tumorigenesis. Methods B-27, growth factors EGF and bFGF was added into DMEM/F-12 culture in serum-free stem cell culture medium for U87 SLCs. Suspended culture of U87 SLCs was suspended using the neuro-sphere formation assay, while adherent culture of U87 SLCs was achieved by coating Matrigel matrix on the culture surface. The mRNA and protein level of stemness biomarkers in culture were detected using real-time quantitative PCR and Western blot. The proportion of CD133+ cells in culture was detected by flow cytometry. The changes of cell oxygen consumption rate were detected by Seahorse cell metabolism analysis. Cell tumorigenesis ability was verified by subcutaneous tumor transplantation in animals. Results U87 SLCs in stem cell culture medium would grow into typical sphere morphology within one week, and the spheres would continue to grow as the culture process prolongs. At the appropriate concentration of adhesive, U87 SLCs adhered to and grow well in stem cell culture medium. The mRNA transcription of stemness biomarkers such as CD133, nestin, OLIG2, CD44, CD15, and integrin α6(ITGA6) was significantly increased as found in both culture methods, and the protein levels of CD133 and nestin were also increased under both methods(P＜0.05). U87 SLCs showed higher mitochondrial reserve respiratory capacity (P＜0.05). U87 SLCs could form larger subcutaneous tumors with fewer inoculated cells (P＜0.05), and grew faster in vivo with stronger tumorigenic ability. Conclusions U87 SLCs have typical stemness characteristics and may function as tumor cell model with higher stemness properties

Passive Resistor Temperature Compensation for a High-Temperature Piezoresistive Pressure Sensor

The main limitation of high-temperature piezoresistive pressure sensors is the variation of output voltage with operating temperature, which seriously reduces their measurement accuracy. This paper presents a passive resistor temperature compensation technique whose parameters are calculated using differential equations. Unlike traditional experiential arithmetic, the differential equations are independent of the parameter deviation among the piezoresistors of the microelectromechanical pressure sensor and the residual stress caused by the fabrication process or a mismatch in the thermal expansion coefficients. The differential equations are solved using calibration data from uncompensated high-temperature piezoresistive pressure sensors. Tests conducted on the calibrated equipment at various temperatures and pressures show that the passive resistor temperature compensation produces a remarkable effect. Additionally, a high-temperature signal-conditioning circuit is used to improve the output sensitivity of the sensor, which can be reduced by the temperature compensation. Compared to traditional experiential arithmetic, the proposed passive resistor temperature compensation technique exhibits less temperature drift and is expected to be highly applicable for pressure measurements in harsh environments with large temperature variations

DNA methylation landscape reveals GNAS as a decitabine-responsive marker in patients with acute myeloid leukemia

Background: The demethylation agent decitabine (DAC) is a pivotal non-intensive alternative treatment for acute myeloid leukemia (AML). However, patient responses to DAC are highly variable, and predictive biomarkers are warranted. Herein, the DNA methylation landscape of patients treated with a DAC-based combination regimen was compared with that of patients treated with standard chemotherapy to develop a molecular approach for predicting clinical response to DAC. Methods: Twenty-five non-M3 AML patients were enrolled and subjected to DNA methylation sequencing and profiling to identify differentially methylated regions (DMRs) and genes of interest. Moreover, the effects of a DAC-based regimen on apoptosis and gene expression were explored using Kasumi-1 and K562 cells. Results: Overall, we identified 541 DMRs that were specifically responsive to DAC, among which 172 DMRs showed hypomethylation patterns upon treatment and were aligned with the promoter regions of 182 genes. In particular, GNAS was identified as a critical DAC-responsive gene, with in vitro GNAS downregulation leading to reduced cell apoptosis induced by DAC and cytarabine combo treatment. Conclusions: We found that GNAS is a DAC-sensitive gene in AML and may serve as a prognostic biomarker to assess the responsiveness of patients with AML to DAC-based therapy