GmDof41 regulated by the DREB1-type protein improves drought and salt tolerance by regulating the DREB2-type protein in soybean.

Despite their essential and multiple roles in biological processes, the molecular mechanism of Dof transcription factors (TFs) for responding to abiotic stresses is rarely reported in plants. We identified a soybean Dof gene GmDof41 which was involved in the responses to drought, salt, and exogenous ABA stresses. Overexpression of GmDof41 in soybean transgenic hairy roots attenuated H2O2 accumulation and regulated proline homeostasis, resulting in the drought and salt tolerance. Yeast one-hybrid and electrophoretic mobility shift assay (EMSA) illustrated that GmDof41 was regulated by the DREB1-type protein GmDREB1B;1 that could improve drought and salt tolerance in plants. Further studies illustrated GmDof41 can directly bind to the promoter of GmDREB2A which encodes a DREB2-type protein and affects abiotic stress tolerance in plants. Collectively, our results suggested that GmDof41 positively regulated drought and salt tolerance by correlating with GmDREB1B;1 and GmDREB2A. This study provides an important basis for further exploring the abiotic stress-tolerance mechanism of Dof TFs in soybean.

A soybean EF-Tu family protein GmEF8, an interactor of GmCBL1, enhances drought and heat tolerance in transgenic Arabidopsis and soybean.

A soybean elongation factor Tu family (EF-Tu) protein, GmEF8, was determined to interact with GmCBL1, and GmEF8 expression was found to be induced by various abiotic stresses such as drought and heat. An ortholog of GmEF8 was identified in Arabidopsis, a T-DNA knockout line for which exhibited hypersensitivity to drought and heat stresses. Complementation with GmEF8 rescued the sensitivity of the Arabidopsis mutant to drought and heat stresses, and GmEF8 overexpression conferred drought and heat tolerance to transgenic Arabidopsis plants. In soybean, plants with GmEF8-overexpressing hairy roots (OE-GmEF8) exhibited enhanced drought and heat tolerance and had higher proline levels compared to plants with RNAi GmEF8-knockdown hairy roots (MR-GmEF8) and control hairy roots (EV). A number of drought-responsive genes, such as GmRD22 and GmP5CS, were induced in the OE-GmEF8 line compared to MR-GmEF8 and EV under normal growth conditions. These results suggest that GmEF8 has a positive role in regulating drought and heat stresses in Arabidopsis and soybean. This study reveals a potential role of the soybean GmEF8 gene in response to abiotic stresses, providing a foundation for further investigation into the complexities of stress signal transduction pathways.

Oxidized low-density lipoprotein receptor 1: a novel potential therapeutic target for intracerebral hemorrhage.

Oxidized low-density lipoprotein receptor 1 (OLR1) is upregulated in neurons and participates in hypertension-induced neuronal apoptosis. OLR1 deletion exerts protective effects on cerebral damage induced by hypertensive-induced stroke. Therefore, OLR1 is likely involved in the progress of intracerebral hemorrhage. In this study, we examined the potential role of OLR1 in intracerebral hemorrhage using a rat model. OLR1 small interfering RNA (10 µL; 50 pmol/µL) was injected into the right basal ganglia to knock down OLR1. Twenty-four hours later, 0.5 U collagenase type VII was injected to induce intracerebral hemorrhage. We found that knockdown of OLR1 attenuated neurological behavior impairment in rats with intracerebral hemorrhage and reduced hematoma, neuron loss, inflammatory reaction, and oxidative stress in rat brain tissue. We also found that silencing of OLR1 suppressed ferroptosis induced by intracerebral hemorrhage and the p38 signaling pathway. Therefore, silencing OLR1 exhibits protective effects against secondary injury of intracerebral hemorrhage. These findings suggest that OLR1 may be a novel potential therapeutic target for intracerebral hemorrhage.

[Induction and Anti-Tumor Function of Tertiary Lymphoid Organs].

OBJECTIVE: To induce the development of tertiary lymphoid organs (TLO) in a mouse model of melanoma and to evaluate TLO's functions in antitumor immunity. METHODS: Lymphotoxin-beta receptor (LTßR) was overexpressed in NIH3T3 cells through the lentivirus system and the overexpression efficiency of LTßR in LTßR-NIH3T3 cells was examined. Western blot and qPCR were used to examine the non-canonical nuclear factor (NF)-κB signaling pathway in NIH3T3 cells overexpressing LTßR. B16-OVA melanoma mouse model was constructed to explore the induction of TLO and anti-tumor functions of TLO in LTßR-NIH3T3 cells. RESULTS: LTßR was overexpressed in NIH3T3 cells through the lentivirus system, and flow cytometry showed that the proportion of GFP + cells reached 99%. The overexpression of LTßR activated the non-canonical NF-κB signaling pathway in NIH3T3 cells. Findings from the mouse tumor model suggest that the injection of LTßR-NIH3T3 cells successfully induced the development of lymphoid tissue around the tumor and enhanced the tumor infiltration of T cells and MHCⅡ + macrophages, significantly inhibiting tumor growth and prolonging the survival of tumor-bearing mice. CONCLUSION: LTßR-NIH3T3 cells promoted anti-tumor immunity by inducing TLO development, which may provide new perspectives for tumor immunotherapy.

Mechanisms Underlying the Antidepressant Effect of Acupuncture via the CaMK Signaling Pathway.

The CaMK pathway has been proven to play an important role in regulating cognitive function and emotional response. Acupuncture through the CaMK pathway improves depression-like behavior and the molecular mechanism related to its antidepressant remains to be explored. In this study, we aimed to determine whether the ability of acupuncture at Baihui (GV20) and Shenting (GV24) points to treat depression is related to the regulation of key proteins in the CaMK pathway. A rat model of depression was induced by chronic unpredicted mild stress (CUMS). Model rats in the electroacupuncture group were subjected to acupuncture at the Baihui (GV20) and Shenting (GV24) acupoints once a day for 20 min. Model rats in the fluoxetine group were gavaged with fluoxetine (1.8 mg/kg). Immunohistochemistry and Western blotting assays were used to evaluate immunoreactivity for and the protein expression levels of CaMKII, CaMKIV, and CaM. The results showed that electroacupuncture had a significant effect in rats with depression. Electroacupuncture and fluoxetine regulated the expression of key proteins in the CaMK signaling pathway, which is related to depression, in the hippocampi of rats. This indicates that acupuncture at Baihui (GV20) and Shenting (GV24) may alleviate depressive symptoms and reduce work- and life-related burdens and stress by regulating the CaMK signaling pathway.

MicroRNA-181c provides neuroprotection in an intracerebral hemorrhage model.

Apoptosis is an important factor during the early stage of intracerebral hemorrhage. MiR-181c plays a key regulatory role in apoptosis. However, whether miR-181c is involved in apoptosis of prophase cells after intracerebral hemorrhage remains unclear. Therefore, in vitro and in vivo experiments were conducted to test this hypothesis. In vivo experiments: collagenase type VII was injected into the basal ganglia of adult Sprague-Dawley rats to establish an intracerebral hemorrhage model. MiR-181c mimic or inhibitor was injected in situ 4 hours after intracerebral hemorrhage. Neurological functional defects (neurological severity scores) were assessed 1, 7, and 14 days after model establishment. Terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick-end labeling and western blot assay were conducted 14 days after model establishment. In vitro experiments: PC12 cells were cultured under oxygen-glucose deprivation, and hemins were added to simulate intracerebral hemorrhage in vitro. MiR-181c mimic or inhibitor was added to regulate miR-181c expression. 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay, luciferase reporter system, and western blot assay were performed. Experimental results revealed differences in miR-181c expression in brain tissues of both patients and rats with cerebral hemorrhage. In addition, in vitro experiments found that miR-181c overexpression could upregulate the Bcl-2/Bax ratio to inhibit apoptosis, while inhibition of miR-181c expression could reduce the Bcl-2/Bax ratio and aggravate apoptosis of cells. Regulation of apoptosis occurred through the phosphoinositide 3 kinase (PI3K)/Akt pathway by targeting of phosphatase and tensin homolog deleted on chromosome ten (PTEN). Higher miR-181c overexpression correlated with lower neurological severity scores, indicating better recovery of neurological function. In conclusion, miR-181c affects the prognosis of intracerebral hemorrhage by regulating apoptosis, and these effects might be directly mediated and regulated by targeting of the PTEN\PI3K/Akt pathway and Bcl-2/Bax ratio. Furthermore, these results indicated that miR-181c played a neuroprotective role in intracerebral hemorrhage by regulating apoptosis of nerve cells, thus providing a potential target for the prevention and treatment of intracerebral hemorrhage. Testing of human serum was authorized by the Ethics Committee of China Medical University (No. 2012-38-1) on February 20, 2012. The protocol was registered with the Chinese Clinical Trial Registry (Registration No. ChiCTR-COC-17013559). The animal study was approved by the Institutional Animal Care and Use Committee of China Medical University (approval No. 2017008) on March 8, 2017.

Survey of Machine Learning Techniques for Prediction of the Isoform Specificity of Cytochrome P450 Substrates.

BACKGROUND: Determination or prediction of the Absorption, Distribution, Metabolism, and Excretion (ADME) properties of drug candidates and drug-induced toxicity plays crucial roles in drug discovery and development. Metabolism is one of the most complicated pharmacokinetic properties to be understood and predicted. However, experimental determination of the substrate binding, selectivity, sites and rates of metabolism is time- and recourse- consuming. In the phase I metabolism of foreign compounds (i.e., most of drugs), cytochrome P450 enzymes play a key role. To help develop drugs with proper ADME properties, computational models are highly desired to predict the ADME properties of drug candidates, particularly for drugs binding to cytochrome P450. OBJECTIVE: This narrative review aims to briefly summarize machine learning techniques used in the prediction of the cytochrome P450 isoform specificity of drug candidates. RESULTS: Both single-label and multi-label classification methods have demonstrated good performance on modelling and prediction of the isoform specificity of substrates based on their quantitative descriptors. CONCLUSION: This review provides a guide for researchers to develop machine learning-based methods to predict the cytochrome P450 isoform specificity of drug candidates.

Passive Transmembrane Permeation Mechanisms of Monovalent Ions Explored by Molecular Dynamics Simulations.

Passive or unassisted ion permeation through lipid bilayers involves a type of rare events by which cells regulate their salt concentrations and pH. It is important to understand its mechanism in order to develop technologies of, for example, delivering or maintaining small drug-like molecules inside cells. In earlier simulations of passive ion permeations, the commonly used sampling methods usually define the positions of ions relative to the membrane as a measure of permeation, i.e., the collective variable, ignoring the active participations of other particles. Newly defined collective variables involving the movements of ions, lipids, and water molecules allow us to identify the transition paths on the free energy landscape using the 2D umbrella sampling techniques. In this work, this technique was used to study the permeation processes of some well-known ions, sodium, potassium, and chloride. It is found permeations of sodium and potassium are assisted by important lipid bilayer deformations and massive water solvation, while chloride may not. Chloride may have two different possible pathways, in which the energetic favorable one is similar to the solubility-diffusion model. The free energy barriers for the permeation of these ions are in semiquantitative agreement with experiments. Further analyses on the distributions of oxygens and interaction energies suggest the electrostatic interactions between ions and polar headgroups of lipids may greatly influence membrane deformation as well as the water wire and furthermore the free energy barriers of waterwire mediated pathways. For chloride, the nonwaterwire pathway may be energetically favorable.

The dependence of structural stability and tunable gap on the Si components of ZnSe/Si bi-coaxial nanowire heterostructures.

The bare and hydrogen-passivated ZnSe/Si bi-coaxial nanowire heterostructures along [110] direction have been investigated by using the first-principle calculations within density functional theory. The structural stability and electronic property of ZnSe/Si bi-coaxial nanowire heterostructures have been shown by changing the Si components. It is found that the ZnSe/Si nanowires have zero gaps at lower Si components, and then they have the increasing gap at higher Si components. It is seen clearly that there is the transition of band gap form zero to nonzero. With increasing Si components, the ZnSe/Si nanowires can be also achieved as n-type or p-type, in agreement qualitatively with the experimental observations. In addition, the structural stabilities and the cohesive energies of ZnSe/Si bi-coaxial nanowires are changed obviously with the different Si components.

[Spatial discharge characteristics and total load control of non-point source pollutants based on the catchment scale].

Agricultural non-point source pollution is one of the major causes of water quality deterioration. Based on the analysis of the spatial discharge characteristics and intensity of major pollutants from the agricultural pollution source, the establishment of spatial management subzones for controlling agricultural non-point pollution and a design of a plan for total load control of pollutants from each subzone is an important way to improve the efficiency of control measures. In this paper the Four Lake basin in Hubei Province is adopted as the research case region and a systematic research of the control countermeasures of agricultural non-point pollution based on the catchment scale is carried out. The results shows that in the Four Lake basin, the COD, total nitrogen, total phosphorus and ammonia nitrogen load of the water environment are mainly caused by agricultural non-point pollution. These four kinds of non-point source pollutants respectively account for 67.6%, 82.2%, 84.7% and 50.9% of the total pollutant discharge amount in the basin. The analysis of the spatial discharge characteristics of non-point source pollutants in the Four Lake basin shows that the major contributor source regions of non-point source pollutant in the basin are the four counties, including Honghu, Jianli, Qianjiang and Shayang where the aquatic and livestock production are relatively developed. According to the spatial discharge characteristics of the pollutants and the evaluation of the discharge intensity of pollutants, the Four Lake basin is divided into three agricultural non-point pollution management subzones, which including Changhu upstream aquatic and livestock production pollution control subzone, Four-lake trunk canal rural non-point source pollution control subzone and Honghu aquatic production pollution control subzone. Specific pollution control measures are put forward for each subzone. With a comprehensive consideration of the water quality amelioration and the allowable discharge of pollutants, a total load control solution is designed for the three non-point pollution management subzones, so as to fulfill the requirements of all indices of the monitoring sites and the requirements for the allowable discharge of pollutants of the water. Among the major pollutants, the major COD reduction area includes the Four-lake trunk canal subzone and the Honghu Lake subzone, which respectively account for 43% and 42% of the total COD reduction amount; the major ammonia nitrogen reduction area includes the Four-lake trunk canal subzone, accounting for 66% of the total ammonia nitrogen reduction amount; the major total nitrogen reduction area covers the Four-lake trunk canal subzone and the Honghu Lake subzone, accounting for 42% and 31% of the total nitrogen reduction amount in the basin respectively; the major total phosphorus reduction area is the Four-lake trunk canal subzone, accounting for 53% of the total phosphorus reduction amount in the basin.

[Distribution and accumulation of mineral nitrogen in apple orchard soils in dry plateau of eastern Gansu Province].

This paper studied the distribution and accumulation characteristics of mineral nitrogen in the soils of different years old apple orchards in dry plateau of eastern Gansu Province. The soil ammonium nitrogen content was increased with the increase of planting year. In the 2-3, 5, 10, 15, 20, and 22 years old apple orchards, the ammonium nitrogen content in 0-120 cm soil layer was 3.3, 5.8, 6.5, 9.1, 12.1, and 15.3 mg x kg(-1), respectively, and 0-60 cm soil layer had a higher ammonium nitrogen content than 60-120 cm soil layer. For all the apple orchards, the nitrate nitrogen content in 0-40 cm soil layer was relatively low, but increased with increasing depth. The soil nitrate nitrogen content also increased with the increase of planting year, which reached 2602.5 kg x hm(-2) in the 0-120 cm soil layer of 22 years old apple orchard. It was concluded that in the apple orchards in dry plateau of eastern Gansu Province, soil ammonium nitrogen was more accumulated in upper layers, while nitrate nitrogen was more accumulated in deeper layers.

Identification of 12 animal species meat by T-RFLP on the 12S rRNA gene.

The verification of authenticity of meat products is relevant for economical, religious or public health concerning reasons. A molecular approach using terminal restriction fragment length polymorphism (T-RFLP) was developed to distinguish 12 common economically important meat species. The partial 12S rRNA gene was amplified with double-fluorescently labeled primers. The amplified fragments were digested with two endonucleases and only the terminal restriction fragment containing labeled primer was detected on capillary electrophoresis system ABI3100. AluI and Tru9I generated differently-sized terminal fragments in different species. Pig and buffalo can be separated by 3'-terminal fragment of AluI digestion. Horse, turkey, goat, sheep, deer, and cattle can be further separated by 5'-terminal fragment of Tru9I digestion. Dog and chicken, sturgeon and salmon can finally be separated by 5'-terminal fragment of AluI digestion and 3'-terminal fragment of Tru9I digestion. Our results demonstrated the potential feasibility and applicability of T-RFLP method for rapid and accurate identification of animal species.

[Biological control efficiency of Amblyseius cucumeris (Oudemans) on Panonychus ulmi (Koch)].

Based on the life-table parameters and the release effects of Amblyseius cucumeris (Oudemans) under laboratory and field conditions, the control efficiency of A. cucumeris (Oudemans) on Panonychus ulmi (Koch) was evaluated. When feeding on P. ulmi, A. cucumeris could complete its development period and lay eggs with one generation time of 9.54 d at (25 +/- 1) degrees C. The egg-laying period (24.85 d) and lifespan (38.52 d) of female A. cucumeris were longer than their preys (15.93 d and 34.79 d, respectively), but the average amount of eggs laid by female A. cucumeris was 36.70, being 43.9% less than that of P. ulmi. The net reproductive rate (R0) and intrinsic rate of natural increase (r(m)) of P. ulmi were 103.9% and 13.2% higher than those of A. cucumeris, respectively. No matter what the good-harm ratios was 1 : 30 or 1 : 150, the control efficiency of A. cucumeris after its 20-day release under laboratory condition could be over 95%, and the efficiency after 60-day release under field condition could be 91.73%, being 23.77% higher than that when sprayed with pesticide, suggesting that A. cucumeris was a good biological agent to control the population growth of P. ulmi. When the P. ulmi population density was less than 2 per leaf, releasing A. cucumeris in the orchard could effectively control the population increase of P. ulmi.