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Abstract A single load compression test of rock-coal-rock assemblages (RCRs) containing coal bodies with different inclinations was carried out with the research background of mining deep, large-steep coal seams. It was found that the larger the inclination angle of the coal body in the RCR samples, the larger the compaction stage of the samples and the smaller the uniaxial compressive strength value. In addition, both the maximum acoustic emission (AE) energy of the samples and the cumulative AE energy at the moment of destruction decreased with the increase of the inclination angle of the coal body in the form of exponential relativities. Also, the input energy and ultimate elastic energy of the samples at the peak stress moment decreased with increased inclination angle of the coal body. Furthermore, both of them changed with the inclination angle of the coal body in accordance with the exponential relationship; meanwhile, when the applied load exceeded the peak strength, the dissipative energy of the RCR samples increased rapidly due to the loss of load-bearing capacity. At the peak moment, the percentage of dissipated energy of the samples increased exponentially with the increase of the inclination angle of the coal body. This study has certain reference significance for the prevention and control of dynamic disasters during the mining of large angle coal seams.

The distribution of the deep geothermal field and the tectonic thermal control mode in the Yingshan area were not understood systematically, which would bring in constraints to the sustainable exploration of local geothermal resources. Based on the field survey on continuous temperature, fracture measurements in the outcrops of granite rock, slug-test, and pumping test in the boreholes, a three-dimensional model coupling fluid flow and heat transfer process in fractured rock reservoir was established. Then the distribution of fluid pressure temperature and the Darcy velocity in the whole geothermal reservoir were calculated considering different treatment combinations of main faults. The results show that the deep temperature field and fluid pressure distribution are greatly affected by the faults system. At the location of bedrock and impermeable fault, the fluid movement is very weak, and the heat transfer is mainly controlled by heat conduction, while in the high permeable fault, it is mainly controlled by heat convection. Low temperature and fluid pressure occurred in the deep, high-permeable fault zone whereas high fluid pressure occurred in the impermeable fault zone. At a depth larger than 2000 m, the zones with high temperatures and intense hydraulic exchange coincide with the conductive fault zones, which indicates that the high-permeable faults are the most important channel for seepage and heat transfer in the reservoir. Therefore, the northeast faults are the dominant channel for fluid migration and heat conduction in the Yinshan area. Hot water migrates upward along the northeast faults and is blocked by the north-south fault. The hot spring is formed at the intersecting position of two groups of faults. This study is helpful for scientific, sustainable exploration of geothermal resources in the Yingshan area.

To clarify the evolution of thermal maturity and pore structure in continental organic-rich shales, calcareous shales of the Liaohe Basin (China) were pyrolyzed, and examined using Raman spectroscopy, Fourier transform infrared spectroscopy (FTIR), X-ray diffraction (XRD), X-ray photoelectron spectroscopy (XPS), scanning electron microscopy (SEM), nitrogen sorption isotherms, and nuclear magnetic resonance (NMR) spectroscopy. The increase in Raman G‒D band separation and decrease in Raman ID/IG ratio with increasing thermal maturity indicate that these parameters provide superior thermal maturity indicators. This is also confirmed by the good linear correlation of G‒D band shifts and ID/IG with vitrinite reflectance (VR) and maximum temperature (Tmax), respectively. The relative detection accuracy (DA), sampling requirements (SR), sample preparation (SP), detection time (DT), and equipment requirement (ER) of VR, G‒D band shifts, ID/IG, Tmax, FTIR, and XPS indicate that Raman analysis is a simple, quick, and robust method to evaluate thermal maturity. The moderate SR, complex SP, and long DT suggest that VR and Tmax are less widely applicable for characterizing thermal maturity. The FTIR and XPS techniques provide semi-quantitative maturity indicators with poor DA and high ER. Pores observed within organic matter and minerals under SEM indicate that an increase in thermal maturity not only affects the development of organic pores but is also beneficial for the formation of mineral dissolution pores, such as those formed during the transformation of Na0.95Ca0.16Al1.16Si2.84O8 to Na0.84Ca0.02Al1.02Si2.98O8, a process confirmed by XRD. The BET and NMR data also indicate that the development of pore structure is closely related to the evolution of thermal maturity in calcareous shale. During the initial stage, primary pores are filled by bitumen generated from kerogen; this leads to a decrease in transition pores, mesopores, and shale porosity, and reduced pore connectivity. Then, secondary nanoscale pores, transition pores, and mesopores increase with increasing thermal maturity. The peak in secondary porosity is consistent with the liquid hydrocarbon production rate peak, a process that increases shale porosity and leads to improved pore connectivity. The dissolution of minerals induced by organic acids may also contribute to this secondary porosity. With a further increase in thermal maturity, secondary porosity at the microscale is further developed, while transition pores and mesopores collapse, resulting in reduced pore connectivity. The poor pore connectivity that occurs at both low and high VR values may be more conducive to the preservation of shale oil and gas. This study is significant for research into the evolution of thermal maturity and pore structure in continental organic-rich shales.

Abstract Biochar and manure can be used for sustainable land management. However, little is known about how soil amendments might affect surface and belowground microbial processes and subsequent wood decomposition. In a split‐split‐split plot design, we amended soil with two rates of manure (whole plot; 0 and 9 Mg ha−1) and biochar (split plot; 0 and 10 Mg ha−1). Wood stakes of three species (hybrid poplar, triploid Populus tomentosa Carr.; aspen, Populus tremuloides Michx.; and pine, Pinus taeda L.) were placed in two positions (horizontally on the soil surface, and inserted vertically in the mineral soil), which served as a substrate for fungal growth. In 3 years, the decomposition rate (density loss), moisture content, and fungal community (via high‐throughput sequencing methods) of stakes were evaluated. Results indicated that biochar and/or manure increased the wood stake decomposition rates, moisture content, and operational taxonomic unit abundance. However, the richness and diversity of fungi were dependent on wood stake position (surface > mineral), species (pine > the two Populus), and sample dates. This study highlights that soil amendment with biochar and/or manure can alter the fungal community, which in turn can enhance an important soil process (i.e., decomposition).

Abstract While biochar and manure can provide considerable benefits to soil properties, how these amendments may alter soil microbial activity and decomposition processes remains unknown. In a split‐split‐split‐plot experiment, we amended a sandy loam soil with three rates of manure (whole plot; 0, 3, 9 Mg ha−1) and biochar (split‐plot; 0, 2.5, 10 Mg ha−1), and installed three species of wood stakes (split‐split‐split plot; triploid poplar, Populus tomentosa Carr.; trembling aspen, Populus tremuloides Michx.; and loblolly pine, Pinus taeda L.) on the soil surface and in the mineral soil (split‐split plot) to serve as a substrate for microbial degradation. Wood stakes were sampled 3 years after installation to assess decomposition rates (mass loss), and changes in wood carbon (C) and nitrogen (N). In addition, soil extracellular enzyme activities at the 0–20 cm depth were examined. Biochar alone, especially 10 Mg ha−1, increased wood stake decomposition and moisture content on the soil surface and in the mineral soil. Manure at the rate of 9 Mg ha−1 increased soil N‐acetyl‐β‐D‐glucosaminidase, α‐glucosidase, and aryl sulfatase activities by 91%, 17%, and 48% respectively. Because of the synergistic benefits of biochar and manure, we suggest that, in this climatic regime and soil texture, 10 Mg ha−1 biochar can be used for soil C sequestration and soil quality improvement, and 9 Mg ha−1 manure can be used in combination with biochar to build soil organic matter in plantations.

Polyaniline (PANI) has long been explored as a promising organic cathode for Li-ion batteries. However, its poor electrochemical utilization and cycling instability cast doubt on its potential for practical applications. In this work, we revisit the electrochemical performance of PANI in nonaqueous electrolytes, and reveal an unprecedented reversible capacity of 197.2 mAh g−1 (244.8 F g−1) when cycled in a wide potential range of 1.5 to 4.4 V vs. Li+/Li. This ultra-high capacity derives from 70% -NH- transformed to =NH+- during deep charging/discharging process. This material also demonstrates a high average coulombic efficiency of 98%, an excellent rate performance with 73.5 mAh g−1 at 1800 mA g−1, and retains 76% of initial value after 100 cycles, which are among the best reported values for PANI electrodes in battery applications.

Vaccines are one of the most effective means of preventing influenza A, typically containing the hemagglutinin (HA) of the influenza A virus. However, antigenic drift and shift of the influenza A virus can lead to instability in vaccine efficacy. Compared to HA, the antigenic variation rate of neuraminidase (NA) is slower. In traditional inactivated influenza vaccines, although they contain a certain amount of NA, there are significant differences between different batches, which cannot consistently induce NA-based immune responses. Therefore, NA is often overlooked in vaccine development. In this study, we report an mRNA vaccine encoding the NA of two strains of influenza A virus. The experimental results demonstrated that when matched with the viral strain, this mRNA vaccine induced high levels of neutralizing antibodies, providing a protective effect to mice in viral challenge experiments, and this immune response was shown to be biased towards the Th1 type. In summary, this study demonstrates that NA is a promising potential antigen, providing new insights for the development of influenza A virus vaccines.

Summary: Background: In a previous phase 3 clinical trial, we showed that an inactivated poliovirus vaccine derived from the Sabin strain (sIPV) can induce neutralising antibodies against currently circulating and reference wild poliovirus strains. However, the immune persistence of sIPV remains to be evaluated. Methods: In this study, 400 participants who were eligible for an early phase 3 clinical trial (Jan 1, 2012–Aug 31, 2014) in Pingle County, GuanXi Province, China, were initially involved in one site. Of the participants in the previous phase 3 clinical trial, sera of 287, 262, 237, and 207 participants were sampled at the ages of 4, 6, 8, and 10 years, respectively, after the prime-boost regimen. Neutralising antibodies against attenuated Sabin strains were detected using these serum samples to determine immune persistence. The serum neutralising antibodies titre of 1:8 against poliovirus types 1, 2, and 3 is considered to be a seroprotection level for polio. The trial is registered at ClinicalTrials.gov, NCT01510366. Findings: The protective rates against poliovirus types 1, 2, and 3 in the sIPV group were all 100% at 10 years after the booster immunisation, compared with 98.1%, 100%, and 97.1%, respectively, in the wIPV control group after 10 years. After the booster at 18 months, the geometric mean titres (GMTs) of neutralising antibodies against poliovirus types 1, 2, and 3 in the sIPV group were 13,265.6, 7856.7, and 6432.2, respectively, and the GMTs in the control group (inoculated with inactivated poliovirus vaccine derived from wild strain (wIPV)) were 3915.6, 2842.6, and 4982.7, respectively. With increasing time after booster immunisation, the GMTs of neutralising antibodies against poliovirus types 1, 2, and 3 gradually decreased in both the sIPV and wIPV groups. At the age of ten years, the GMTs of neutralising antibodies against poliovirus types 1, 2, and 3 in the sIPV group were 452.3, 392.8, and 347.5, respectively, and the GMTs in the wIPV group 108.5, 154.8, and 229.3, respectively, which were still at a higher-than-protective level (1:8). Interpretation: Both sIPV and wIPV maintained sufficiently high immune persistence against poliovirus types 1, 2, and 3 for at least 10 years after booster immunisation. Funding: Yunnan Provincial Science and Technology Department, the Bill and Melinda Gates Foundation, the National High-tech Research and Development Program, the National International Science and Technology Cooperation Project, the Yunnan Application Basic Research Project, the Innovation Team Project of Xie He, the Yunnan International Scientific and Technological Cooperation Project, and the Medical and Technology Innovation Project of Xie He.

Hepatocellular carcinoma (HCC) is now a common cause of cancer death, with no obvious change in patient survival over the past few years. Although the traditional therapeutic modalities for HCC patients mainly involved in surgery, chemotherapy, and radiotherapy, which have achieved admirable achievements, challenges are still existed, such as drug resistance and toxicity. The emerging gene therapy of clustered regularly interspaced short palindromic repeat/CRISPR-associated nuclease 9-based (CRISPR/Cas9), as an alternative to traditional treatment methods, has attracted considerable attention for eradicating resistant malignant tumors and regulating multiple crucial events of target gene-editing. Recently, advances in CRISPR/Cas9-based anti-drugs are presented at the intersection of science, such as chemistry, materials science, tumor biology, and genetics. In this review, the principle as well as statues of CRISPR/Cas9 technique were introduced first to show its feasibility. Additionally, the emphasis was placed on the applications of CRISPR/Cas9 technology in therapeutic HCC. Further, a broad overview of non-viral delivery systems for the CRISPR/Cas9-based anti-drugs in HCC treatment was summarized to delineate their design, action mechanisms, and anticancer applications. Finally, the limitations and prospects of current studies were also discussed, and we hope to provide comprehensively theoretical basis for the designing of anti-drugs.

Zinc metal is a promising anode material for aqueous secondary batteries. However, the unfavourable morphologies formed on the electrode surface during cycling limit its application. Here, the authors report the tailoring of the surface morphology using a lanthanum nitrate aqueous electrolyte additive.

Salt precipitation is an important issue for CO2 sequestration in deep aquifers.At present, most numerical simulations ignore the feedback effect of the changes in the porosity and permeability caused by the salt precipitation on fluid flow.In this paper, taking Liujiagou Formation in Ordos Basin as an example, a two-dimensional model is established using TOUGH2 code.By modifying the source code of the program, the model can consider the feedback effect of salt precipitation on fluid flow.The simulation results show that after the CO2 injection into the Liujiagou Formation for 20 years, salt precipitation makes the formation pressure near the injection well increase by 0.87MPa and the corresponding injectivity loss of the formation is 7.17%.The salinity has the greatest influence on salt precipitation and its feedback, followed by CO2 injection rate, and permeability has the least influence.When the salinity is high, solid salt saturation increases significantly, resulting in a great decrease in the permeability.In this study, when the salinity was 0.24, the salt precipitation caused an injectivity loss by 45.32% and the pressure buildup up to 12.14 MPa.Therefore, special attention should be paid to the salt precipitation caused by high salinity and its feedback effect during CO2 storage.

ObjectiveTo develop a radiomics nomogram for predicting microvascular invasion (MVI) before surgery in hepatocellular carcinoma (HCC) patients.Materials and MethodsThe data from a total of 189 HCC patients (training cohort: n = 141; validation cohort: n = 48) were collected, involving the clinical data and imaging characteristics. Radiomics features of all patients were extracted from hepatobiliary phase (HBP) in 15 min. Least absolute shrinkage selection operator (LASSO) regression and logistic regression were utilized to reduce data dimensions, feature selection, and to construct a radiomics signature. Clinicoradiological factors were identified according to the univariate and multivariate analyses, which were incorporated into the final predicted nomogram. A nomogram was developed to predict MVI of HCC by combining radiomics signatures and clinicoradiological factors. Radiomics nomograms were evaluated for their discrimination capability, calibration, and clinical usefulness.ResultsIn the clinicoradiological factors, gender, alpha-fetoprotein (AFP) level, tumor shape and halo sign served as the independent risk factors of MVI, with which the area under the curve (AUC) is 0.802. Radiomics signatures covering 14 features at HBP 15 min can effectively predict MVI in HCC, to construct radiomics signature model, with the AUC of 0.732. In the final nomogram model the clinicoradiological factors and radiomics signatures were integrated, outperforming the clinicoradiological model (AUC 0.884 vs. 0.802; p

The flat peach is a high economic value table fruit possessing excellent quality and a unique aroma. This article investigated the quality characteristics and aroma fingerprinting of flat peaches (Qingpan, QP; Ruipan 2, R2; Ruipan 4, R4; Wanpan, WP) from Xinjiang in terms of taste, antioxidant capacity, and volatile aroma compounds using high-performance liquid chromatography (HPLC) and HS-SPME-GC-MS. The results showed that the flat peaches had a good taste and high antioxidant capacity, mainly due to the high sugar–low acid property and high levels of phenolic compounds. This study found that sucrose (63.86~73.86%) was the main sugar, and malic acid (5.93~14.96%) and quinic acid (5.25~15.01%) were the main organic acids. Furthermore, chlorogenic acid (main phenolic compound), epicatechin, rutin, catechin, proanthocyanidin B1, and neochlorogenic acid were positively related to the antioxidant activity of flat peaches. All flat peaches had similar aroma characteristics and were rich in aromatic content. Aldehydes (especially benzaldehyde and 2-hexenal) and esters were the main volatile compounds. The aroma fingerprinting of flat peaches consisted of hexanal, 2-hexenal, nonanal, decanal, benzaldehyde, 2,4-decadienal, dihydro-β-ionone, 6-pentylpyran-2-one, 2-hexenyl acetate, ethyl caprylate, γ-decalactone, and theaspirane, with a “peach-like”, “fruit”, and “coconut-like” aroma. Among them, 2,4-decadienal, 2-hexenyl acetate, and theaspirane were the characteristic aroma compounds of flat peaches. The results provide a theoretical basis for the industrial application of the special aroma of flat peaches.

Nickel-rich LiNi0.8Co0.1Mn0.1O2 (NCM811) is one of the most promising positive electrodes for utilization in the next-generation of lithium-ion batteries. Charging the NCM cells above 4.3 V is proposed to be beneficial for its reversible capacity. However, the high reactivity of the NCM811 usually results in parasitic electrolyte degradation, which is accelerated with the increase of the Ni content in the NCM positive electrodes, leading to the thickening of the positive electrode-electrolyte interphase during cycling. Herein, to counter this issue, we select partially fluorinated solvents, such as methyl 3,3,3-trifluoropropanoate (MTFP) and 1,1,1,2,2,3,3,4,4-nonafluoro-4-methoxybutane (NFMB), as a co-solvent for fluoroethylene carbonate (FEC)-based electrolytes, and detailed investigate theirs physical, chemical, and electrochemical properties for applications in NCM811 materials. Compared to the carbonate-based electrolyte without a fluorinated solvent, the electrolytes with a fluorinated co-solvent display an obviously enhanced cycling performance of the Li/NCM811 cells charged to above 4.5 V. This work suggests that fluorinated co-solvent electrolytes provide an alternative way to the high-concentration electrolyte for the design of new electrolyte systems for high energy density lithium-ion batteries.

The white-spotted bamboo shark (Chiloscyllium plagiosum) is an ecologically and commercially important marine cartilaginous fish in the Indo-Western Pacific. However, the resources of this species are declining in recent years due to habitat degradation and overfishing. Studies on genomics and genetic markers can provide valuable information for sound management and conservation of C. plagiosum. In this study, genome resequencing data of a domesticated C. plagiosum individual were generated for genomic comparison between wild and domesticated samples. Based on 281.17 Gb of clean sequencing data, the genome size of the domesticated sample was estimated as 4.99 Gb, with heterozygosity of 0.51% and repeat ratio of 74.67%, which is about 1 Gb larger than that of the wild sample (3.85 Gb with a repeat ratio of 63.53%). By using a reference-based approach, we assembled a nearly 3.45 Gb genome sequence of the domesticated sample, with the scaffold N50 of 69.45 Mb. Subsequent identification of genome-wide microsatellite markers confirmed the different abundance of repeat elements in wild and domesticated C. plagiosum. Additionally, a total of 8,703,211 single-nucleotide polymorphisms (SNPs) were detected and annotated. The demographic analysis based on identified SNPs revealed a large and constant effective population size of C. plagiosum after the last population expansion (~0.3 million years ago). The genomic data and identified genetic markers in this study can provide fundamental and useful information for further comparative genomics, evolutionary biology, and conservation genetics of C. plagiosum.

Flat peaches possess characteristic flavors and are rich in nutrients. The fermentation of flat peaches to produce wine through complex biochemical reactions is an effective method to overcome their seasonal defects. Spontaneously fermented flat peach wine has plentiful and strong flavors, but the microbiota of fermentation are still unknown. In this study, the microbial succession and volatile compound dynamics of spontaneous fermentation in Xinjiang flat peach wine were investigated using high-throughput sequencing (HTS) and headspace solid phase microextraction (HS-SPME) coupled with gas chromatography-mass spectrometry (GC-MS) technology, respectively, to better understand the microbiota involved. Multivariate data analysis was used to predict the relationship between microorganisms and volatile chemicals. The results showed that Kazachstania, Pichia, Aspergillus, Fructobacillus, Leuconostoc, and Lactobacillus were the dominant genera during the spontaneous fermentation of flat peach wine. Furthermore, ethyl hexanoate, 3-hexen-1-yl acetate, ethyl caprate, ethyl caprylate, phenethyl acetate, ethanol, γ-decalactone, decanal, 1-hexanoic acid, and octanoic acid endued flat peach wine with a strong fruity and fatty aroma. The core functional microbiota (primarily consisting of 11 bacterial and 14 fungal taxa) was strongly associated with the production of 27 volatile compounds in the spontaneously fermented flat peach wine, according to multivariate data analysis. Some alcohols and esters were positively linked with the presence of Kazachstania and Pichia. Meanwhile, the presence of Fructobacillus, Leuconostoc, Lactobacillus, and Weissella was significantly correlated with 2-non-anol, ethanol, 3-methyl-1-butanol, octyl formate, isoamyl lactate, and ethyl lactate. This snapshot of microbial succession and volatile compound dynamics provides insights into the microorganisms involved in flat peach wine fermentation and could guide the production of flat peach wine with desirable characteristics.

Ursolic acid (UA), a natural pentacyclic triterpenoid, possesses widespread biological and pharmacological activities. However, drawbacks such as low bioavailability, poor targeting and rapid metabolism greatly hinder its further clinical application. Recently, with the development of nanotechnology, various UA nanosystems have emerged as promising strategies for effective cancer therapy. This article reviews various types of UA-based nano-delivery systems, primarily with emphasis placed on novel UA-based carrier-free nano-drugs, which are considered to be innovative methods for cancer therapy. Moreover, this review presents carrier-free nano-drugs that co-assembled of UA and photosensitizers that displayed synergistic antitumor performance. Finally, the article also describes the development and challenges of UA nanosystems for future research in this field. Overall, the information presented in this review will provide new insight into the rational utilization of nano-drugs in cancer therapy.

Biochar and manure may be used to enhance soil quality and productivity for sustainable agriculture and forestry operations. However, the response of surface and belowground wood decomposition (i.e., soil processes) and nutrient flux to soil amendments is unknown, and more site-specific information about soil property responses is also essential. In a split-plot design, the soil was amended with three rates of manure (whole plot; 0, 3, and 9 Mg ha−1) and three rates of biochar (split-plot; 0, 2.5, and 10 Mg ha−1). Soil physical properties, nutrients, and enzyme activities were evaluated in two years. In addition, wood stakes of three species (poplar, triploid Populus tomentosa Carr.; aspen, Populus tremuloides Michx.; and pine, Pinus taeda L.) were installed both horizontally on the soil surface and vertically in the mineral soil to serve as an index of soil abiotic and biotic changes. Wood stake mass loss, nitrogen (N), phosphorus (P), and potassium (K) flux were tested. The high rate of both manure and biochar increased soil water content by an average of 18%, but the increase in total soil P, K, organic carbon (C) content, and enzyme activities were restricted to single sample dates or soil depths. Wood stakes decomposed faster according to stake location (mineral > surface) and species (two Populus > pine). On average, soil amendments significantly increased the mass loss of surface and mineral stakes by 18% and 5%, respectively, and it also altered wood stake nutrient cycling. Overall, the decomposition of standard wood stakes can be a great indicator of soil quality changes, and 10 Mg ha−1 of biochar alone or combined with 9 Mg ha−1 of manure can be used for long-term carbon sequestration in plantations with similar soil conditions to the present study.

Somatic embryogenesis (SE) is a process of somatic cells that dedifferentiate to totipotent embryonic stem cells and generate embryos in vitro. Despite recent scientific headway in deciphering the difficulties of somatic embryogenesis, the overall picture of key genes, pathways, and co-expression networks regulating SE is still fragmented. Therefore, deciphering the molecular basis of somatic embryogenesis of hybrid sweetgum remains pertinent. In the present study, we analyzed the transcriptome profiles and gene expression regulation changes via RNA sequencing from three distinct developmental stages of hybrid sweetgum: non-embryogenic callus (NEC), embryogenic callus (EC), and redifferentiation. Comparative transcriptome analysis showed that 19,957 genes were differentially expressed in ten pairwise comparisons of SE. Among these, plant hormone signaling-related genes, especially the auxin and cytokinin signaling components, were significantly enriched in NEC and EC early. The K-means method was used to identify multiple transcription factors, including HB-WOX, B3-ARF, AP2/ERF, and GRFs (growth regulating factors). These transcription factors showed distinct stage- or tissue-specific expression patterns mirroring each of the 12 superclusters to which they belonged. For example, the WOX transcription factor family was expressed only at NEC and EC stages, ARF transcription factor was expressed in EC early, and GRFs was expressed in late SE. It was noteworthy that the AP2/ERF transcription factor family was expressed during the whole SE process, but almost not in roots, stems and leaves. A weighted gene co-expression network analysis (WGCNA) was used in conjunction with the gene expression profiles to recognize the genes and modules that may associate with specific tissues and stages. We constructed co-expression networks and revealed 22 gene modules. Four of these modules with properties relating to embryonic potential, early somatic embryogenesis, and somatic embryo development, as well as some hub genes, were identified for further functional studied. Through a combination analysis of WGCNA and K-means, SE-related genes including AUX22, ABI3, ARF3, ARF5, AIL1, AIL5, AGL15, WOX11, WOX9, IAA29, BBM1, MYB36, LEA6, SMR4 and others were obtained, indicating that these genes play an important role in the processes underlying the progression from EC to somatic embryos (SEs) morphogenesis. The transcriptome information provided here will form the foundation for future research on genetic transformation and epigenetic control of plant embryogenesis at a molecular level. In follow-up studies, these data could be used to construct a regulatory network for SE; Key genes obtained from coexpression network analysis at each critical stage of somatic embryo can be considered as potential candidate genes to verify these networks.

Conifers are the world's major source of timber and pulpwood and have great economic and ecological value. Currently, little research on the application of CRISPR/Cas9, the commonly used genome-editing tool in angiosperms, has been reported in coniferous species. An efficient CRISPR/Cas9 system based on somatic embryogenesis (SEis) suitable for conifers could benefit both fundamental and applied research in these species. In this study, the SpCas9 gene was optimized based on codon bias in white spruce, and a spruce U6 promoter was cloned and function-validated for use in a conifer specific CRISPR/Cas9 toolbox, i.e., PgCas9/PaU6. With this toolbox, a genome-editing vector was constructed to target the DXS1 gene of white spruce. By Agrobacterium-mediated transformation, the genome-editing vector was then transferred into embryogenic tissue of white spruce. Three resistant embryogenic tissues were obtained and used for regenerating plants via SEis. Albino somatic embryo (SE) plants with mutations in DXS1 were obtained in all of the three events, and the ratios of the homozygous and biallelic mutants in the 18 albino mutants detected were 22.2% in both cases. Green plants with mutations in DXS1 were also produced, and the ratios of the DXS1 mutants to the total green plants were 7.9, 28, and 13.5%, respectively, among the three events. Since 22.7% of the total 44 mutants were edited at both of the target sites 1 and 2, the CRISPR/Cas9 toolbox in this research could be used for multi-sites genome editing. More than 2,000 SE plants were regenerated in vitro after genome editing, and part of them showed differences in plant development. Both chimerism and mosaicism were found in the SE plants of white spruce after genome editing with the CRISPR/Cas9 toolbox. The conifer-specific CRISPR/Cas9 system developed in this research could be valuable in gene function research and trait improvement.

Cryopreservation is one of the key technologies for the mass propagation of conifers via somatic embryogenesis. Cryotolerance and embryogenecity of conifer somatic embryos (SEs) could be affected by different temperature treatments, for which the underlying mechanisms were unknown. In this study, the developing SEs of Picea glauca obtained their cryotolerance with a survival rate of 100% when cultured on maturation medium at either 23°C for 4 weeks or 4°C for 10 weeks. However, only the embryos that underwent 4°C acclimation remained high embryogenicity, i.e., 91.7% based on cryovials or 29.3% on the plant tissue. Analysis of differentially expressed genes (DEGs) revealed that both 23 and 4°C treatments led to drastic changes in the gene expression, i.e., 21,621 and 14,906 genes, respectively, and the general increase in many oligosaccharides and flavonoids, in addition to the content change of proline (1.9- and 2.3-fold at 23 or 4°C) and gallic acid (6,963- and 22,053-fold). There were 249 significantly different metabolites between the samples of 23 and 4°C treatments and the changing trend of the sorbitol, fatty acids, and monosaccharides differed between these samples. During 4°C-acclimation, the metabolites of the arginine biosynthesis pathway increased between 2.4- and 8.1-fold, and the expression of antioxidant genes was up-regulated significantly. At 4°C, the up-regulated genes were for germ-like proteins, instead of seed storage proteins at 23°C. Concentrations of abscisic acid and jasmonic acid increased up to 2- and 1.5-fold, respectively, in the cold-acclimated embryos. After 10 weeks at 4°C, the embryos stayed at pre-cotyledonary stage with 17.1% less DNA methylation and fewer storage substances than those at 23°C for 4 weeks, which developed cotyledons. This research provides new insights into mechanisms underlying the response of SEs to different culture temperatures and benefits method development for germplasm conservation in conifers.

Globally, the red wine market experienced a rapid growth in the last decade, due to the superior colour, taste, and nutritional quality. The red grapes used for vinification have individual characteristics varying within the regional environment. In this study, the quality of seven grape cultivars, including Marselan, Yan 73, Muscat Hamburg, Kadarka, Merlot, Cabernet Sauvignon, and Crimpose, and their corresponding wines, were investigated based on high-performance liquid chromatography and headspace solid-phase microextraction coupled to gas chromatography–mass spectrometry. These techniques were performed to analyze the chemical compositions and volatile compounds of the tested samples, respectively. The results showed that tartaric acid (29.96% to 73.45%) and rutin (12.53% to 56.54%) were the dominant organic acid and phenolic compounds in grapes, respectively. Higher concentrations of organic acids and phenolic compounds, and the types of volatile compounds, were observed to be highest in the Cabernet Sauvignon grape. The antioxidant activity of wines ranged from 6.74 to 102.68 mmol TE/L, and Yan 73 wine had the highest antioxidant activity. A total of 69 volatile compounds consisting of 17 alcohols, 26 esters, 5 aldehydes, 9 acids, 7 ketones, and 5 other volatile compounds were identified in all tested wines, and 11 important aroma active substances (odor activity value > 1) were selected, consisting of β-ionone, phenethyl acetate, geranyl acetate, ethyl 9-decenoate, ethyl caprate, ethyl pelargonate, decanal, ethyl caprylate, 6-methyl-5-hepten-2-one, methyl 2-hexenoate, and ethyl hexanoate, which endow wines with a unique aroma. This work clearly describes the chemical and sensory characteristics of seven red grape cultivars in Xinjiang of China and provides diversity options for cultivars for winemaking.

Doctors’ diagnosis preferences are different, which makes them adopt different assumptions in medical decision making. Taking the diagnosis of thyroid nodules as an example, this study compares three assumptions, namely deletion, imputation based on the distribution (distribution), and benign by default (benign). For deletion, which is the most used assumption, the clinical reports with missing features would be deleted. For distribution, the missing features would be replaced with a distribution of features with respective probabilities. Besides the two assumptions, certain doctors have also stated that they leave benign features unrecorded because they think that such benign features are irrelevant to the final diagnosis. Under the benign assumption, the missing features would be replaced with benign features. The three assumptions are tested comparatively. Moreover, the belief rule base (BRB) is used to construct the diagnostic model under the three assumptions since it is essentially a white-box approach that can provide good interpretability and direct access to doctors and patients. A total of 3766 clinical reports on thyroid nodule diagnosis were collected from ten radiologists over a seven-year period. Case study results validate that the benign by default assumption has produced the optimal results, although different doctors could present varied tendencies towards different assumptions. Guidance and suggestions for doctors’ practical work have been made based on the study results to improve work efficiency and diagnostic accuracy.

Peaches are tasty and juicy, with a unique flavor. The flavors of peaches always vary with cultivars. To investigate the physicochemical and aroma characteristics of peaches, the sugars, organic acids, total flavonoids, phenols, antioxidant activities, and aroma compounds of seven peach cultivars in Xinjiang were determined using high-performance liquid chromatography (HPLC) and headspace solid-phase microextraction coupled with gas chromatography–mass spectrometry (HS-SPME–GC–MS). The results showed that sucrose (59.83 to 87.34%), malic acid (32.41 to 59.14%), and chlorogenic acid (10.43 to 45.50%) were the dominant sugar, organic acid, and phenolic compound in peaches, respectively. The antioxidant activity varied between 147.81 and 394.55 μmol TEs/100 g. The analysis of the aroma structure of peaches found that the volatile composition of peaches was relatively consistent, though the concentration of total aroma and certain separate compounds were different between cultivars. Meanwhile, the aroma fingerprint of the peaches consisted of hexyl acetate, cis-3-hexenyl acetate, γ-decalactone, n-hexanal, 2-hexenal, nonanal, decanal benzaldehyde and 6-pentylpyran-2-one, providing a clear green, sweet, floral, and fruity odor. These results provide complete information on the physicochemical properties, functional ingredients and aroma of the peaches.

Abstract Background High temperature is a major environmental stress that limits plant growth and agriculture productivity. Mitogen-activated protein kinases (MAPKs) are highly conserved serine and threonine protein kinases that participate in response to diverse environmental stresses in plants. A total of 16 putative SlMAPK genes are identified in tomato, and SlMAPK3 is one of the most extensively studied SlMAPKs. However, the role of SlMAPK3 in response to heat stress is not clearly understood in tomato plants. In this study, we performed functional analysis of SlMAPK3 for its possible role in response to heat stress. Results qRT-PCR analyses revealed that SlMAPK3 relative expression was depressed by heat stress. Here, wild-type (WT) tomato plants and CRISPR/Cas9-mediated slmapk3 mutant lines (L8 and L13) were used to investigate the function of SlMAPK3 in response to heat stress. Compared with WT plants, slmapk3 mutants exhibited less severe wilting and less membrane damage, showed lower reactive oxygen species (ROS) contents, and presented higher both activities and transcript levels of antioxidant enzymes, as well as elevated expressions of genes encoding heat stress transcription factors (HSFs) and heat shock proteins (HSPs). Conclusions CRISPR/Cas9-mediated slmapk3 mutants exhibited more tolerance to heat stress than WT plants, suggesting that SlMAPK3 was a negative regulator of thermotolerance. Moreover, antioxidant enzymes and HSPs/HSFs genes expression were involved in SlMAPK3-mediated heat stress response in tomato plants.

Abstract Background NPR1, nonexpressor of pathogenesis-related gene 1, is a master regulator involved in plant defense response to pathogens, and its regulatory mechanism in the defense pathway has been relatively clear. However, information about the function of NPR1 in plant response to abiotic stress is still limited. Tomato is the fourth most economically crop worldwide and also one of the best-characterized model plants employed in genetic studies. Because of the lack of a stable tomato NPR1 (SlNPR1) mutant, little is known about the function of SlNPR1 in tomato response to biotic and abiotic stresses. Results Here we isolated SlNPR1 from tomato ‘Ailsa Craig’ and generated slnpr1 mutants using the CRISPR/Cas9 system. Analysis of the cis-acting elements indicated that SlNPR1 might be involved in tomato plant response to drought stress. Expression pattern analysis showed that SlNPR1 was expressed in all plant tissues, and it was strongly induced by drought stress. Thus, we investigated the function of SlNPR1 in tomato-plant drought tolerance. Results showed that slnpr1 mutants exhibited reduced drought tolerance with increased stomatal aperture, higher electrolytic leakage, malondialdehyde (MDA) and hydrogen peroxide (H2O2) levels, and lower activity levels of antioxidant enzymes, compared to wild type (WT) plants. The reduced drought tolerance of slnpr1 mutants was further reflected by the down-regulated expression of drought related key genes, including SlGST, SlDHN, and SlDREB. Conclusions Collectively, the data suggest that SlNPR1 is involved in regulating tomato plant drought response. These results aid in further understanding the molecular basis underlying SlNPR1 mediation of tomato drought sensitivity.

Molten-salt assisted solid-state synthesis is considered a promising method in obtaining layer-structured cathodes for lithium-ion batteries with homogeneous elemental distribution and controllable morphologies. However, drawbacks like resource wasting which have arisen from redundant salt removal after sintering greatly hinder the broader application of this technique. In this paper, a closed-loop utilization of molten salts is proposed for the first time with specific electrochemical properties of the products studied. Results confirm the feasibility of this strategy, and that with only replenishing LiOH, the recycled LiOH-LiCl molten salts can be successfully reused into another LiNi0.5Co0.2Mn0.3O2 synthesis, which exhibits almost equal electrochemical performance to the product using fresh molten salts. We believe this research can provide significant insight in guiding green and environmental-friendly preparation methods involving molten salts.

The effects of CO2-water-rock interactions on the injectivity and safety of CO2 geological storage have drawn wide attention. The geochemical reaction mechanisms in carbonate formations after CO2 injection are still controversial. To better understand the transformation of injected CO2 in carbonates and the involved geochemical reactions, we first conducted autoclave experiments reproducing the in-situ conditions of the Lianglitage Formation, Yingshan Formation, and Qiulitage Formation at the Tazhong Uplift in the Tarim Basin. We then established a batch model using TOUGHREACT-ECO2H, validated with the experimental results, to simulate the long-term CO2-water-rock interactions. It was found that the initial mineral compositions and water chemistry have important effects on the CO2-water-rock interactions in carbonate formations. The experiment results show that the dissolution of calcite and dolomite dominates in the early reaction period. However, we still observed some secondary minerals, such as ankerite, montmorillonite, calcite, and dolomite. The CO2-water-rock reactions can be more dramatic when the contents of calcite and dolomite in carbonates are closer. Moreover, the long-term simulation results show that calcite, magnesite, and hematite are the main formed secondary minerals, whereas dolomite is the major dissolved mineral. This study is helpful for a better understanding of the CO2 mineral trapping mechanism in carbonate formations.

Pseudotsuga gaussenii is a forest species under protection in China. Propagation via somatic embryogenesis is efficient and needed for recovering this species. In this research, effects of medium supplements, i.e., methylglyoxal-bis (guanylhydrazone) (MGBG), polyethylene glycol (PEG), and maltose, on somatic embryo (SE) development were studied. More mature SEs developed with the pre-maturation treatment medium supplemented with 50 μM or 100 μM MGBG. During SE maturation, when the medium was supplemented with both PEG and maltose, the over-proliferation of embryogenic tissue was suppressed. When maltose was eliminated from the maturation medium, SEs were slightly smaller but developed better than other treatments due to the supply of lower carbon source, whereas when PEG was taken out from the maturation medium, no mature SE was obtained, only the heavily-proliferated plant tissue. Meanwhile, DNA methylation levels differed in these SE maturation cultures at the late culture stage of SE maturation. The level increased sharply with no maltose treatment (20.28%) while decreasing without PEG supplement (10.35%). DNA methyltransferase genes MET1-1, MET1-2, MET1-3, CMT3, DRM1, and DRM2 were partially cloned in this study to detect their expression level via qPCR. Expression of these genes, except MET1-3, in the culture of no PEG was higher significantly than the cultures of other treatments at weeks 1, 3, and 10. Our results suggested that MGBG and/or PEG play an important role in stimulating SE development and maturation. Furthermore, sugar supplements at a lower level benefited SE maturation in Pseudotsuga gaussenii.

In conifers, somatic embryogenesis is uniquely initiated from immature embryos in a narrow time window, which is considerably hindered by the difficulty to induce embryogenic tissue (ET) from other tissues, including mature somatic embryos. In this study, the embryogenic ability of newly induced ET and DNA methylation levels was detected, and whole-transcriptome sequencing analyses were carried out. The results showed that ultra-low temperature treatment significantly enhanced ET induction from mature somatic embryos, with the induction rate from 0.4% to 15.5%, but the underlying mechanisms remain unclear. The newly induced ET showed higher capability in generating mature embryos than the original ET. DNA methylation levels fluctuated during the ET induction process. Here, WGCNA analysis revealed that OPT4, TIP1-1, Chi I, GASA5, GST, LAX3, WRKY7, MYBS3, LRR-RLK, PBL7, and WIN1 genes are involved in stress response and auxin signal transduction. Through co-expression analysis, lncRNAs MSTRG.505746.1, MSTRG.1070680.1, and MSTRG.33602.1 might bind to pre-novel_miR_339 to promote the expression of WRKY7 genes for stress response; LAX3 could be protected by lncRNAs MSTRG.1070680.1 and MSTRG.33602.1 via serving as sponges for novel_miR_495 to initiate auxin signal transduction; lncRNAs MSTRG.505746.1, MSTRG.1070680.1, and MSTRG.33602.1 might serve as sponges for novel_miR_527 to enhance the expression of Chi I for early somatic embryo development. This study provides new insight into the area of stress-enhanced early somatic embryogenesis in conifers, which is also attributable to practical applications.

Picture fuzzy sets, which are the extension of intuitionistic fuzzy sets, can deal with inconsistent information better in practical applications. A distance measure is an important mathematical tool to calculate the difference degree between picture fuzzy sets. Although some distance measures of picture fuzzy sets have been constructed, there are some unreasonable and counterintuitive cases. The main reason is that the existing distance measures do not or seldom consider the refusal degree of picture fuzzy sets. In order to solve these unreasonable and counterintuitive cases, in this paper, we propose a dynamic distance measure of picture fuzzy sets based on a picture fuzzy point operator. Through a numerical comparison and multi-criteria decision-making problems, we show that the proposed distance measure is reasonable and effective.

Ta-doped Li7La3Zr2O12 (LLZTO) solid-state electrolytes (SEs) show great promise for solid-state batteries due to its high conductivity and safety. However, one of the challenges it faces is lithium dendrite propagation upon long-term cycling. To address this issue, we propose the incorporation of fumed silica (FS) at the grain boundaries of LLZTO to modify the properties of the garnet pellet, which effectively inhibits the dendrite growth. The introduction of FS has demonstrated several beneficial effects. Firstly, it reduces the migration barrier of lithium ions, which helps prevent dendrite formation and propagation. Additionally, FS reduces the electronic conductivity of the SEs pellet, suppressing the dendrite formation. Moreover, the formed lithium silicates from FS might also be acted as electron inhibitor, thus inhibiting the lithium dendrite growth upon cycling. By investigating the use of FS as a modifier in LLZTO-based electrolytes, our study contributes to advancing dendrite-free solid-state electrolytes and thus the development of high-performance all-solid-state batteries. [ABSTRACT FROM AUTHOR]