Correlation of bilateral M1 hand area excitability and overall functional recovery after spinal cord injury: protocol for a prospective cohort study.

BACKGROUND: After spinal cord injury (SCI), a large number of survivors suffer from severe motor dysfunction (MD). Although the injury site is in the spinal cord, excitability significantly decreases in the primary motor cortex (M1), especially in the lower extremity (LE) area. Unfortunately, M1 LE area-targeted repetitive transcranial magnetic stimulation (rTMS) has not achieved significant motor improvement in individuals with SCI. A recent study reported that the M1 hand area in individuals with SCl contains a compositional code (the movement-coding component of neural activity) that links matching movements from the upper extremities (UE) and the LE. However, the correlation between bilateral M1 hand area excitability and overall functional recovery is unknown. OBJECTIVE: To clarify the changes in the excitability of the bilateral M1 hand area after SCI and its correlation with motor recovery, we aim to specify the therapeutic parameters of rTMS for SCI motor rehabilitation. METHODS: This study is a 12-month prospective cohort study. The neurophysiological and overall functional status of the participants will be assessed. The primary outcomes included single-pulse and paired-pulse TMS. The second outcome included functional near-infrared spectroscopy (fNIRS) measurements. Overall functional status included total motor score, modified Ashworth scale score, ASIA Impairment Scale grade, spinal cord independence measure and modified Barthel index. The data will be recorded for individuals with SCI at disease durations of 1 month, 2 months, 4 months, 6 months and 12 months. The matched healthy controls will be measured during the same period of time after recruitment. DISCUSSION: The present study is the first to analyze the role of bilateral M1 hand area excitability changes in the evaluation and prediction of overall functional recovery (including motor function and activities of daily living) after SCI, which will further expand the traditional theory of the predominant role of M1, optimize the current rTMS treatment, and explore the brain-computer interface design for individuals with SCI. TRIAL REGISTRATION NUMBER: ChiCTR2300068831.

Microbial mechanism of biochar addition to reduce N2O emissions from soilless substrate systems.

The soilless peat-based substrate partially solves the global soil problem in greenhouse vegetable production. However, it still produces serious N2O emissions due to the application of nutrient solutions. The pyrolysis biochar is regarded as an effective measure to reduce soil N2O emissions. However, the effect and mechanism of biochar on N2O emissions from the soilless substrate remain unknown. Therefore, this study set up six treatments by adjusting the ratio of biochar addition of peat-based substrate: 0% (0BC), 2% (2BC), 4% (4BC), 6% (6BC), 8% (8BC) and 10% (10BC) (v/v). The results showed that compared to the control treatment, N2O emissions reduced by 81%, 71%, 51%, 61%, and 75% in the 2BC, 4BC, 6BC, 8BC and 10BC treatments, respectively. In addition, lettuce yield increased by 10% and 7% in the 2BC and 4BC treatments and decreased by 0.5%, 4% and 6% in the 6BC, 8BC and 10BC treatments, respectively. Combining stable isotope technology, qPCR analysis and high-throughput sequencing, five microbial pathways of N2O production, including bacterial and archaea nitrification (BN and AN), denitrification performed by fungi, denitrifier bacteria and nitrifier bacteria (FD, DD and ND), were roughly distinguished. In addition, the extent of N2O reduction was obtained by δ18O vs.δ15NSP map. For all treatments, overall, the DD process (over 50%) was the main process of N2O production and reduction, while ND and AN processes were almost negligible (less 5%). In detail, the decrease of N2O emissions was caused by decreasing the contribution of FD in the 6BC, 8BC and 10BC treatments and reducing the contribution of BN in the 0BC and 2BC treatments. In addition, biochar addition increased the extent of N2O reduction to N2. In summary, the 2% biochar addition presented the greatest extent of N2O reduction to N2 (83%) and the lowest N2O emissions as well as the highest lettuce yields and nitrogen utilization efficiency. Therefore, 2% biochar is deemed the most optimal addition to the peat-based substrate.

Evaluation of N2O sources after fertilizers application in vegetable soil by dual isotopocule plots approach.

Nitrogen (N) fertilizer is the major deriver of nitrous oxide (N2O) emissions in agricultural soil. In the vegetable fields in China both inorganic and organic fertilizers are largely applied as basic sources of nitrogen. Identifying the effects of fertilizer type on soil microbial activities involved in N2O emissions would be of great help for future development of N2O reduction strategies. N2O isotopocule deltas, including δ15Nbulk, δ18O and SP (the 15N site preference in N2O), have been used to analyze microbial pathways of N2O production under different treatments, including bio-organic fertilizer treatment, half bio-organic fertilizer and half urea (mixed fertilizer) treatment, urea treatment and no fertilizer treatment. We measured environmental factors, N2O fluxes and N2O isotopocule deltas to evaluate the dynamics of N2O emissions and constructed the dual isotopocule plots (δ15Nbulk vs. SP and δ18O vs. SP) of the main N2O emission phases to assess contribution of the involved microbial processes (bacterial nitrification, bacterial denitrification, nitrifier denitrification and fungal denitrification). According to the results of the main N2O emission phases, we found that bio-organic fertilizer and mix fertilizer treatments had significantly lower N2O emissions compared to urea treatment, with average N2O fluxes of 1477 ± 204, 1243 ± 187 and 1941 ± 164 µg m-3 h-1, respectively, but there were no significant effects on mineral N and cabbage yield. In addition, the urea treatment and the mixed fertilizer treatment had close and higher nitrogen use efficiency. Furthermore, the δ18O vs. SP plot was useful for providing insight into microbial processes, showing that fungal denitrification/bacterial nitrification was the dominant microbial pathway and bio-organic fertilizer and mix fertilizer treatments had higher denitrification and N2O reduction compared to urea treatment. Those findings demonstrated that the partial replacement of urea with bio-organic fertilizer was a better choice, by means of enhancing denitrification to reduce N2O emissions and also guaranteeing the nitrogen use efficiency and the cabbage yield.

Relationship between left ventricular vortex and preejectional flow velocity during isovolumic contraction studied by using vector flow mapping.

OBJECTIVES: The purpose of this study was to investigate the relationship between the vortex in left ventricle (LV) during the isovolumic contraction (IVC) period and the preejectional flow velocity in LV outflow tract (VLVOT ). METHODS: Color Doppler loops were acquired for vector flow mapping in apical long-axis view in 76 patients with dilated cardiomyopathy, 61 patients with coronary artery disease and 36 healthy controls. RESULTS: All normals exhibited an IVC vortex reaching the LV base. VLVOT was significantly related to IVC vortex area flux, transmitral A velocity, mitral annular a' velocity and E/e' ratio, respectively. Transmitral A velocity was the only independent predictor of VLVOT (R2  = 0.292, P = 0.001). In patients the IVC vortex could reach the LV base, middle, or apex. VLVOT was significantly related to range, area and area flux of the IVC vortex, LV size, LVEF, mitral annular velocities, E/e' ratio, transmitral A velocity, and IVC time, respectively. Range and corrected area flux of the IVC vortex, LV end-systolic short diameter, and IVC time were independent predictors of VLVOT (R2  = 0.608, P < 0.001). CONCLUSIONS: In normals, the transmitral A velocity (momentum) is efficiently transferred from mitral orifice to LV outflow tract by a normally formed IVC vortex, and transmitral A velocity is the only independent predictor of VLVOT . However, in patients with a wide range of LV enlargement and dysfunction, the momentum transfer is associated with not only the LV dimension and function, but also the range and volume of the IVC vortex.

Factors influencing the end-diastolic vortex assessed by using vector flow mapping.

OBJECTIVES: The purpose of this study was to assess the factors influencing the late diastolic vortex in normal and abnormal ventricles. METHODS: Color Doppler data in left ventricle (LV) were acquired from apical long-axis view and analyzed using vector flow mapping in 57 patients with coronary artery disease, 57 patients with dilated cardiomyopathy, and 53 healthy volunteers. RESULTS: In normals, corrected area and flux of the end-diastolic vortex were positively correlated with transmitral A velocity and heart rate. Subjects with E/A <1 had higher vortex flux than those with E/A >1. Heart rate was the only independent predictor of corrected vortex area (R2  = .170, P = .004), and transmitral A velocity and heart rate were the independent predictors of corrected vortex flux (R2  = .490, P < .001). Patients with various mitral filling patterns showed significant differences in vortex area and flux. The vortex area and flux were positively correlated with transmitral i velocity and a'. Transmitral A velocity was the only independent predictor of corrected vortex area (R2  = .180, P < .001), while transmitral A velocity, heart rate, LV end-systolic short diameter, and end-diastolic long diameter were the independent determinants of corrected vortex flux (R2  = .593, P < .001). CONCLUSIONS: The end-diastolic vortex is formed and mainly affected by the late LV filling. The compensatory atrial contraction may enhance the end-diastolic vortex that facilitates coupling between diastole and systole. LV size can influence the end-diastolic vortex in patients with LV dysfunction and enlargement.

Overexpression of the saccharopine dehydrogenase gene improves lysine biosynthesis in Flammulina velutipes.

Saccharopine dehydrogenase (EC 1.5.1.7) regulates the last step of fungal lysine biosynthesis. The gene (Fvsdh) encoding saccharopine dehydrogenase was identified and cloned from the whole genome of Flammulina velutipes. The genomic DNA of Fvsdh is 1257 bp, comprising three introns and four exons. The full-length complementary DNA of Fvsdh comprises 1107 bp with a deduced amino acid sequence of 368 residues. A 1,000-bp promoter sequence containing the TATA box, CAAT box, and several putative cis-acting elements was also identified. The results of tissue expression analysis showed that the expression level of the Fvsdh gene was higher in the pileus than in the stipe whether in the elongation or maturation stage. Further research showed that the lysine contents were 3.03 and 2.95 mg/g in maturation-pileus and elongation-pileus, respectively. In contrast, the lysine contents were 2.49 and 2.07 mg/g in elongation-stipe and maturation-stipe, respectively. To study the function of Fvsdh, we overexpressed Fvsdh in F. velutipes and found that Fvsdh gene expression was increased from 1.1- to 3-fold in randomly selected transgenic strains. The lysine contents were also increased from 1.12- to 1.3-fold in these five transformants, except for strain T3, in which the lysine contents were the same as the control. These results indicate that the expression of the Fvsdh gene can affect the lysine content of F. velutipes.

Effects of Atmospheric and Room Temperature Plasma (ARTP) Mutagenesis on Physicochemical Characteristics and Immune Activity In Vitro of Hericium erinaceus Polysaccharides.

The polysaccharide is the main active substance contained in Hericium erinaceus and is commonly used in the treatment of neurasthenia, tumors, and digestive diseases. Six intracellular polysaccharide components were obtained from H. erinaceus fruiting bodies cultivated by ARTP (atmospheric and room temperature plasma) mutagenic strain (321) and the original strain (0605), respectively. This study was designed to investigate the physicochemical characteristics of these polysaccharide components and their potential immunomodulatory activities on RAW264.7 macrophages. The results showed that the yield of fruiting body cultivated by mutated strain increased by 22% and the polysaccharide content improved by 16% compared with the original one owing to ARTP mutagenesis. The molecular weight distribution and the monosaccharide compositions of polysaccharide components from H. erinaceus induced by ARTP mutagenesis were significantly different from that of the original one. The NO, IL-6, IL-10, IL-1ß, and TNF-α production activities of macrophages were enhanced by stimulation of 20% ethanol precipitated polysaccharides from H. erinaceus induced by ARTP mutagenesis. These results indicated that ARTP is an efficient and practical method for high polysaccharide content breeding of the H. erinaceus strain and this provided a reference for obtaining high quality resources and healthy product development from H. erinaceus.

Research on the dynamic behavior of flexible drilling tools in ultrashort-radius radial horizontal wells.

A flexible drilling tool is a special drilling tool for ultrashort-radius radial horizontal wells. This tool is composed of many parts and has the characteristics of a multibody system. In this paper, a numerical method for the dynamic analysis of flexible drilling tools is proposed. The flexible drill tool is discretized into spatial beam elements, while the multilayer contact of the flexible drilling tool is represented by the multilayer dynamic gap element, and the dynamic model of the multibody system for the flexible drilling tool's multilayer contact is established, considering the interaction force between the drill bit and the rock. The nonlinear dynamic equation is solved using the Newmark method and Newton-Raphson method. An analysis of the dynamic behavior of a flexible drilling tool is conducted. The results indicate that the flexible drilling tool experiences vortex formation due to the interaction between the flexible drilling pipe and the guide pipe, leading to increased friction and wear. This situation hinders safe drilling operations with flexible drilling tools. The collision force of the flexible drilling tool near the bottom of the hole is more severe than that of the other tool types, which may lead to failure of the connection.

Study on tank damage and response of adjacent tanks in full time domain of detonation.

The flammable gas volatilized from the storage medium in the tank will burn and explode under the action of an accidental ignition source. Tank explosions formed by the explosive shock wave, explosive debris will not only cause their own damage, but also on the safe operation of adjacent tanks to form a threat. Therefore, in this paper, the vertical dome roof tank is taken as the research object, the criteria of brittle fracture failure of tank materials are constructed, and the multi-field coupling calculation model of tank detonation based on TNT equivalent method is established, taking into account the fluid properties inside and outside the tank, the structure of implosion tank and adjacent tank. By analyzing the full time domain process of gas detonation in tanks under different influence factors, the response to implosion damage of tanks and adjacent tanks under the action of explosion shock wave overpressure and debris was obtained. With the increase of the liquid level, the overpressure and deformation of the shock wave of the detonation tank and adjacent tank decrease, and the type and number of fragments decrease. With the increase of the volume of the tank, the overpressure and stress of the shock wave become larger, the strain rate of the tank becomes larger, and brittle failure is more likely to occur. The types and number of fragments formed increase, but detonation fragments are not formed when the tank is full. The research results in this paper provide a theoretical basis and calculation method for preventing and controlling tank implosion accidents, as well as tank design and tank spacing layout.

Influence of leaf inclination angle and tillering on population transpiration, soil evaporation, and yield in winter wheat near-isogenic lines.

Leaf inclination angle (LIA) and tillering impact the winter wheat (Triticum aestivum L.) population canopy structure. Understanding their effects on water use (WU) parameters and yield can guide water-saving strategies through population control. In this study, six near-isogenic lines (NILs) and their parents were selected as materials. These special materials were characterized by varying tillering at the current sowing density, a similar genetic background, and, particularly, a gradient in mean flag leaf LIA. The investigation focused on the jointing to early grain-filling stage, the peak water requirement period of wheat crops. Population-scale transpiration (PT) and evaporation from the soil surface (E) were partitioned from total evapotranspiration (ET) by the means of micro-lysimeters. The results showed decreased PT, E, and ET with increased population density (PD) within a narrow density range derived from varying tillering across genotypes. Significant correlations existed between PD and ET, E, and PT, especially in the wettest 2017-2018 growing season. Within such narrow PD range, all the correlations between WU parameters and PD were negative, although some correlations were not statistically significant, thereby suggesting the population structure's predominant impact. No significant correlation existed between LIA and both ET and PT within the LIA range of 35°-65°. However, significant correlations occurred between LIA and E in two growing seasons. Genotypes with similar LIA but different PD produced varied ET; while with similar PD, the four pairs of genotypes with different LIA each consumed similar ET, thus highlighting PD's more crucial role in regulating ET. The yield increased with higher LIA, and showed a significant correlation, emphasizing the LIA's significant effect on yield. However, no correlation was observed with PD, indicating the minor effect of tillering at the current sowing density. Therefore these results might offer valuable insights for breeding water-saving cultivars and optimizing population structures for effective field water conservation.

Comparative Analysis of Main Agronomic Traits of Different Pleurotus giganteus Germplasm Resources.

Agronomic traits are key components in variety protection, cultivar development, and the formulation of DUS (distinct, uniform, and stable) test guidelines. P. giganteus is an increasingly popular and commercially promising edible macrofungi. In this study, both mycelial performance and fruiting body characters of 15 Pleurotus giganteus strains were investigated. The temperature gradient culture test indicated that, although most of the strains achieved optimal mycelial growth between 24 and 28 °C, a statistical difference in mycelial growth rates and temperature adaptability among strains were found, supporting that this trait has the potential to be adopted as an indicator in distinguishing strains. In the fruiting performance tests, the coefficient of variation (CV) of tested traits ranged from 5.30% (pileus diameter) to 18.70% (individual mushroom weight). The mushroom yields ranged from 103.37 g/bag (strain No. 15) to 275.76 g/bag (strain No. 9). The large divergence observed in individual mushroom weight tested strains, ranging from 40.88 g to 78.39 g (with median between 37.69 and 79.395 g), make it highly selective and a potential indicator in variety development. Strain No. 9 had the advantages of forming larger, heavier fruiting bodies and a more obvious funnel shape, which also exhibited the highest biological efficiency (15.61%). The results suggested some morphological traits showed high variety difference, such as pileus diameter (55.75 mm to 66.48 mm), stipe length (92.59 mm to 177.51 mm), stipe diameter (16.14 mm to 23.52 mm), and pileus thickness (13.38 mm to 19.75 mm). In the cluster analysis, the tested strains were grouped into four clusters based on agronomic traits: cluster Ⅰ comprised six strains (No. 6, No. 11, No. 8, No. 1, No. 14, and No. 9) with high mushroom yield; cluster Ⅱ included four strains (No. 3, No. 10, No. 7, and No. 4) with large pileus diameter and short stipe; cluster ⅡI consisted of four strains (No. 5, No. 12, No. 13, and No. 15) with relatively lower yields; and cluster Ⅳ included only strain No. 2 which was low in yield, individual mushroom weight, and biological efficiency, accompanied by smaller pileus size and shorter stipe. The results of the correlation analysis indicated three traits, including individual mushroom weight, stipe length, and pileus weight, were positively associated with high yield. This study suggested P. giganteus germplasm resources are of high abundance and their agronomic diversity is useful in distinguishing and developing different varieties. The findings of this work provide knowledge on the agronomic traits and cultivation performance of various P. giganteus strains, laying a foundation for the development of its DUS test guidelines and variety protection, as well as providing reference for the breeding and phenotype selection of high-quality cultivars.

Comparative analysis of proteomes and transcriptomes revealed the molecular mechanism of development and nutrition of Pleurotus giganteus at different fruiting body development stages.

Pleurotus giganteus is a commercially cultivated high-temperature mushroom. Investigating the molecular mechanism of fruiting body development will help us to better understand the regulation of substrates and energy in this process. However, little information has been reported on the development and nutrients of the P. giganteus fruiting body. In the present study, P. giganteus is cultivated in a climate chamber, and comparative transcriptome, proteome, and nutritional analysis of P. giganteus fruiting bodies were performed. Our results revealed that Cytochrome P450 monooxygenases and hydrophobin proteins play important roles during the differentiation in the elongation stage. Later, carbon metabolism dominate the fruiting body metabolism and genes related to the carbohydrate metabolic process, glycolytic process, and gluconeogenesis were up-regulated in the mature fruiting bodies. The up-regulation of carbohydrate substrates utilization CAZymes genes and inconsistent protein expression in pileus indicated a reverse transportation of mRNA from the fruiting body to vegetative mycelia. In addition, protein concentration in the pileus is higher than that in the stem, while the stem is the major nitrogen metabolic and amino acid synthetic location. The integrated transcriptomic, proteomic, and nutritional analysis indicated a two-way transportation of substrates and mRNAs in P. giganteus. Stem synthesizes amino acids and transported them to pileus with reducing sugars, while pileus induces the expression of substrate degradation mRNA according to the needs of growth and development and transports them in the other direction.

Insights into production and consumption processes of nitrous oxide emitted from soilless culture systems by dual isotopocule plot and functional genes.

Soilless culture systems (SCS) play an increasing role in greenhouse vegetable production. In the SCS, soilless substrates serve as the major substitute for soil, supplying nutrients to plants but releasing greenhouse gases into the atmosphere. Remarkably, there is a serious problem of N2O emission due to excessive input of N fertilizer. However, the microbial processes of N2O production and consumption in soilless substrates have been rarely studied resulting in difficultly interpreting for its global warming potential. Therefore, these pathways from two classic soilless substrates under two irrigation patterns were investigated by stable isotope technology combined with qPCR analysis in present study. The results according to the dual isotopocule plot of δ15NSP vs. δ18O showed that the mean contribution of denitrification and the mean extent of N2O reduction of case i (Reduction-Mixing) were 26.2 and 81.2 % for the treatment of peat based substrate under drip irrigation (PD), 47.7 and 70.3 % for the treatment of coir substrate under drip irrigation (CD), 29.0 and 80.8 % for the treatment of peat based substrate under tidal irrigation (PT), and 50.8 and 47.4 % for the treatment of coir substrate under tidal irrigation (CT). These results were also further confirmed by the abundance of major functional genes including AOA amoA, nirK and nosZ. Altogether, N2O emission and its microbial processes are determined by substrate types instead of irrigation patterns. For detail, denitrification dominated in the peat based substrate and nitrification dominated in the coir substrate. Compared to the coir substrate, the peat based substrate had higher abundance of functional genes and stronger denitrification and thus generated more N2O. For the two soilless substrates, moreover, the microbiome replaced the mineral N content as the limiting factor for N2O emission. In the SCS, in summary, the two soilless substrates play an important role in tomato growth, but might suffer from inorganic nutrient surplus and microbial shortage. More importantly, the combined analysis of N2O isotopocule deltas and functional genes is a robust tool and provides reliable conclusions for clarifying the microbial processes of N2O production and consumption, thus it is also recommended for use in environments other than soilless substrates.

Responses of the Mushroom Pleurotus ostreatus under Different CO2 Concentration by Comparative Proteomic Analyses.

BACKGROUND: Pleurotus ostreatus is a popular edible mushroom in East Asian markets. Research on the responses of P. ostreatus under different carbon dioxide concentrations is limited. METHODS: Label-free LC-MS/MS quantitative proteomics analysis technique was adopted to obtain the protein expression profiles of P. ostreatus fruiting body pileus collected under different carbon dioxide concentrations. The Pearson correlation coefficient analysis and principal component analysis were performed to reveal the correlation among samples. The differentially expressed proteins (DEPs) were organized. Gene ontology analysis was performed to divide the DEPs into different metabolic processes and pathways. RESULTS: The expansion of stipes was inhibited in the high CO2 group compared with that in the low CO2 group. There were 415 DEPs (131 up- and 284 down-regulated) in P. ostreatus PH11 treated with 1% CO2 concentration compared with P. ostreatus under atmospheric conditions. Proteins related to hydrolase activity, including several amidohydrolases and cell wall synthesis proteins, were highly expressed under high CO2 concentration. Most of the kinases and elongation factors were significantly down-regulated under high CO2 concentration. The results suggest that the metabolic regulation and development processes were inhibited under high CO2 concentrations. In addition, the sexual differentiation process protein Isp4 was inhibited under high CO2 concentrations, indicating that the sexual reproductive process was also inhibited under high CO2 concentrations, which is inconsistent with the small fruiting body pileus under high CO2 concentrations. CONCLUSIONS: This research reports the proteome analysis of commercially relevant edible fungi P. ostreatus under different carbon dioxide concentrations. This study deepens our understanding of the mechanism for CO2-induced morphological change in the P. ostreatus fruiting body, which will facilitate the artificial cultivation of edible mushrooms.

The study of early screening technique for fruiting ability of Lentinula edodes hybrid progenies.

Crossbreeding is the most commonly used method in breeding of Lentinula edodes, however low fruiting rate of the hybrids has always caused troubles and barriers for breeders. An early screening method of the fruiting ability could make the breeding work more efficient. In this paper, a rapid and high-throughput laccase activity detection method based on agar diffusion principle was developed. In this way, we investigated the constitutive and inducible extracellular laccase activity of 36 strains in a breeding population of L. edodes on different media and performed a correlation analysis with fruiting ability of these strains. The results showed the laccase activity of mycelium cultured in non-induced medium for 8 d could be used as an early screening index to judge whether it had fruiting ability at the later stage. Early rapid and simple screening method for hybrid populations was established based on laccase activity characteristics of mycelia. 127 strains from another 5 different hybrid populations were used to verify the early screening method. From the validation results, the early screening method was effective, but the appropriate screening threshold was needed to select according to the cross population, which would greatly to improve the breeding efficiency of L. edodes.

The influence of soil acidification on N2O emissions derived from fungal and bacterial denitrification using dual isotopocule mapping and acetylene inhibition.

Denitrification, as both origins and sinks of N2O, occurs extensively, and is of critical importance for regulating N2O emissions in acidified soils. However, whether soil acidification stimulates N2O emissions, and if so for what reason contributes to stimulate the emissions is uncertain and how the N2O fractions from fungal (ffD) and bacterial (fbD) denitrification change with soil pH is unclear. Thus, a pH gradient (6.2, 7.1, 8.7) was set via manipulating cropland soils (initial pH 8.7) in North China to illustrate the effect of soil acidification on fungal and bacterial denitrification after the addition of KNO3 and glucose. For source partitioning, we used and compared SP/δ18O mapping approach (SP/δ18O MAP) and acetylene inhibition technique combined isotope two endmember mixing model (AIT-IEM). The results showed significantly higher N2O emissions in the acidified soils (pH 6.2 and pH 7.1) compared with the initial soil (pH 8.7). The cumulative N2O emissions during the whole incubation period (15 days) ranged from 7.1 mg N kg-1 for pH 8.7-18.9 mg N kg-1 for pH 6.2. With the addition of glucose, relative to treatments without glucose, this emission also increased with the decrement of pH values, and were significantly stimulated. Similarly, the highest N2O emissions and N2O/(N2O + N2) ratios (rN2O) were observed in the pH 6.2 treatment. But the difference was the highest cumulative N2O + N2 emissions, which were recorded in the pH 7.1 treatment based on SP/δ18O MAP. Based on both approaches, ffD values slightly increased with the acidification of soil, and bacterial denitrification was the dominant pathway in all treatments. The SP/δ18O MAP data indicated that both the rN2O and ffD were lower compared to AIT-IEM. It has been known for long that low pH may lead to high rN2O of denitrification and ffD, but our documentation of a pervasive pH-control of rN2O and ffD by utilizing combined SP/δ18O MAP and AIT-IEM is new. The results of the evaluated N2O emissions by acidified soils are finely explained by high rN2O and enhanced ffD. We argue that soil pH management should be high on the agenda for mitigating N2O emissions in the future, particularly for regions where long-term excessive nitrogen fertilizer is likely to acidify the soils.

Whole-genome sequence of a high-temperature edible mushroom Pleurotus giganteus (zhudugu).

Most of the sequenced wood-rotting edible mushroom produce fruiting body at relatively low temperatures. Little information has been known about the high-temperature wood-rotting mushroom. Here, we performed de novo sequencing and assembly of the genome of a high-temperature edible mushroom Pleurotus giganteus from a monokaryotic strain zhudugu2 using the Illumina and Pac-Bio CLR sequencing technologies. P. giganteus, also known as Zhudugu in China, is a well-known culinary edible mushroom that has been widely distributed and cultivated in China, Southeast Asia, and South Asia. The genome consists of 40.00 Mb in 27 contigs with a contig N50 of 4.384 Mb. Phylogenetic analysis reveals that P. giganteus and other strains in Pleurotus clustered in one clade. Phylogenetic analysis and average nucleotide identity analysis indicated that the P. giganteus genome showed a closer relationship with other Pleurotus species. Chromosome collinearity analysis revealed a high level of collinearity between P. ostreatus and P. giganteus. There are 12,628 protein-coding genes annotated in this monoploid genome. A total of 481 enzymes accounting for 514 carbohydrate-active enzymes (CAZymes) terms were identified in the P. giganteus genome, including 15 laccases and 10 class II peroxidases predicted in the genome, which revealed the robustness of lignocellulose degradation capacity of P. giganteus. The mating-A type locus of P. giganteus consisted of a pair of homeodomain mating-type genes HD1 and HD2. The mating-B type locus of P. giganteus consisted of at least four pheromone receptor genes and three pheromone genes. The genome is not only beneficial for the genome-assisted breeding of this mushroom but also helps us to understand the high-temperature tolerance of the edible mushroom.

A Simple and Efficient CRISPR/Cas9 System Using a Ribonucleoprotein Method for Flammulina filiformis.

CRISPR/Cas9 systems were established in some edible fungi based on in vivo expressed Cas9 and guide RNA. Compared with those systems, the in vitro assembled Cas9 and sgRNA ribonucleoprotein complexes (RNPs) have more advantages, but only a few examples were reported, and the editing efficiency is relatively low. In this study, we developed and optimized a CRISPR/Cas9 genome-editing method based on in vitro assembled ribonucleoprotein complexes in the mushroom Flammulina filiformis. The surfactant Triton X-100 played a critical role in the optimal method, and the targeting efficiency of the genomic editing reached 100% on a selective medium containing 5-FOA. This study is the first to use an RNP complex delivery to establish a CRISPR/Cas9 genome-editing system in F. filiformis. Moreover, compared with other methods, this method avoids the use of any foreign DNA, thus saving time and labor when it comes to plasmid construction.

Influence of life intervention on anxiety, depression, and quality of life of COVID-19 patients: A protocol for systematic review and meta-analysis.

BACKGROUND: Coronavirus disease 2019 (COVID-19) patients suffer from anxiety, depression, and sleep disorder due to isolation treatment and other reasons. Whether life interventions can be an alternative therapy for COVID-19 patients, accompanied with anxiety, depression, and sleep disorder, is controversial. Therefore, we conducted a meta-analysis and systematic review to evaluate the effects of life interventions on anxiety, depression, and sleep disorder in COVID-19 patients to provide some guidance for clinical application. METHODS: The randomized controlled trials related to the life intervention and COVID-19 from inception to February 2021 will be searched. The following databases are our focused areas: the Cochrane Central Register of Controlled Trials, PubMed, MEDLINE, EMBASE, Web of Science, China National Knowledge Infrastructure, Chinese Biomedical Literature Database, and Wan Fang Database. Two investigators would independently screen the literature according to the inclusion and exclusion criteria, extract data, and evaluate the risk of bias in the included studies. Meta-analysis was performed with RevMan 5.3 software. RESULTS: The results will provide a high-quality synthesis of current evidence for researchers in this subject area. CONCLUSION: The conclusion of our study will provide evidence for the judgment of whether life intervention is an effective intervention on COVID-19 patients. PROSPERO REGISTRATION NUMBER: CRD42020199802.

Characterizing Diversity Based on the Chemical and Nutritional Composition of Shiitake Culinary-Medicinal Mushroom Lentinula edodes (Agaricomycetes) Commonly Cultivated in China.

A comparative study was carried out on the chemical composition and nutritional value of six cultivated Lentinula edodes strains that are widely appreciated in China. The results demonstrated that all investigated L. edodes were good sources of protein (14.87%-27.13%), carbohydrates (62.03%-75.56%), and dietary fiber (35.88%-42.49%) and had low ash (5.24%-6.38%) and low fat (0.80%-1.70%) content. There were significant differences among different cultivars. Shenxiang 215 had high crude protein and dietary fiber contents. Potassium was the most abundant mineral element, followed by phosphorus. Different cultivars exhibited distinct fatty acid compositions and free amino acid profiles. Shenxiang 215 had high essential amino acid content. Polyunsaturated fatty acids were predominant in cultivars 0912, Huxiang F4, and Huxiang F2; however, monounsaturated fatty acids were predominant in other strains. Cultivar 0912 had a better mineral and polyunsaturated fatty acid profile. The amino acid profile and protein quality were systematically investigated referring to the latest version of international amino acid reference patterns, including the amino acid score, ratio coefficient of amino acid, ratio coefficient score of amino acid, essential amino acid index, and protein digestibility - corrected amino acid score. The results demonstrated that Shenxiang 18 had better protein quality. These findings provide a reference for breeders to select parents for directional quality breeding.