Biomechanical properties of articular cartilage in different regions and sites of the knee joint: acquisition of osteochondral allografts.

Osteochondral allograft (OCA) transplantation involves grafting of natural hyaline cartilage and supporting subchondral bone into the cartilage defect area to restore its biomechanical and tissue structure. However, differences in biomechanical properties and donor-host matching may impair the integration of articular cartilage (AC). This study analyzed the biomechanical properties of the AC in different regions of different sites of the knee joint and provided a novel approach to OCA transplantation. Intact stifle joints from skeletally mature pigs were collected from a local abattoir less than 8 h after slaughter. OCAs were collected from different regions of the joints. The patella and the tibial plateau were divided into medial and lateral regions, while the trochlea and femoral condyle were divided into six regions. The OCAs were analyzed and compared for Young's modulus, the compressive modulus, and cartilage thickness. Young's modulus, cartilage thickness, and compressive modulus of OCA were significantly different in different regions of the joints. A negative correlation was observed between Young's modulus and the proportion of the subchondral bone (r = - 0.4241, P < 0.0001). Cartilage thickness was positively correlated with Young's modulus (r = 0.4473, P < 0.0001) and the compressive modulus (r = 0.3678, P < 0.0001). During OCA transplantation, OCAs should be transplanted in the same regions, or at the closest possible regions to maintain consistency of the biomechanical properties and cartilage thickness of the donor and recipient, to ensure smooth integration with the surrounding tissue. A 7 mm depth achieved a higher Young's modulus, and may represent the ideal length.

Effect of Moderate Exercise on the Superficial Zone of Articular Cartilage in Age-Related Osteoarthritis.

This study aimed to evaluate the effect of exercise on the superficial zone of the osteoarticular cartilage during osteoarthritis progression. Three-month-old, nine-month-old, and eighteen-month-old Sprague Dawley rats were randomly divided into two groups, moderate exercise and no exercise, for 10 weeks. Histological staining, immunostaining, and nanoindentation measurements were conducted to detect changes in the superficial zone. X-ray and micro-CT were quantitated to detect alterations in the microarchitecture of the tibial subchondral bone. Cells were extracted from the superficial zone of the cartilage under fluid-flow shear stress conditions to further verify changes in vitro. The number of cells and proteoglycan content in the superficial zone increased more in the exercise group than in the control group. Exercise can change the content and distribution of collagen types I and III in the superficial layer. In addition, TGFß/pSmad2/3 and Prg4 expression levels increased under the intervention of exercise on the superficial zone. Exercise can improve the Young's modulus of the cartilage and reduce the abnormal subchondral bone remodeling which occurs after superficial zone changes. Moderate exercise delays the degeneration of the articular cartilage by its effect on the superficial zone, and the TGFß/pSmad2/3 signaling pathways and Prg4 play an important role.

Research on microseismic localization algorithm with global search and local optimization.

A microseismic localization algorithm that combines global search and local optimization is proposed. The Fewer Conditions Trigger Difference (FCTD) objective function of global search and local optimization is constructed, the execution process of the algorithm is described by numerical simulation, and the global search and local optimization microseismic localization algorithm is verified and applied by field data analysis. The results show that: (1) the global search and local optimization methods have fast search speed in the global range, high convergence accuracy and stable localization results in the local range, and high localization accuracy and stability without relying on the velocity model and initial values in the process of search. (2) By comparing the localization results of different localization methods, the global search and local optimization algorithms have better localization results.

Upregulated ribosome pathway plays a key role in HDAC4, improving the survival rate and biofunction of chondrocytes.

Aims: To explore the novel molecular mechanisms of histone deacetylase 4 (HDAC4) in chondrocytes via RNA sequencing (RNA-seq) analysis. Methods: Empty adenovirus (EP) and a HDAC4 overexpression adenovirus were transfected into cultured human chondrocytes. The cell survival rate was examined by real-time cell analysis (RTCA) and EdU and flow cytometry assays. Cell biofunction was detected by Western blotting. The expression profiles of messenger RNAs (mRNAs) in the EP and HDAC4 transfection groups were assessed using whole-transcriptome sequencing (RNA-seq). Volcano plot, Gene Ontology, and pathway analyses were performed to identify differentially expressed genes (DEGs). For verification of the results, the A289E/S246/467/632 A sites of HDAC4 were mutated to enhance the function of HDAC4 by increasing HDAC4 expression in the nucleus. RNA-seq was performed to identify the molecular mechanism of HDAC4 in chondrocytes. Finally, the top ten DEGs associated with ribosomes were verified by quantitative polymerase chain reaction (QPCR) in chondrocytes, and the top gene was verified both in vitro and in vivo. Results: HDAC4 markedly improved the survival rate and biofunction of chondrocytes. RNA-seq analysis of the EP and HDAC4 groups showed that HDAC4 induced 2,668 significant gene expression changes in chondrocytes (1,483 genes upregulated and 1,185 genes downregulated, p < 0.05), and ribosomes exhibited especially large increases. The results were confirmed by RNA-seq of the EP versus mutated HDAC4 groups and the validations in vitro and in vivo. Conclusion: The enhanced ribosome pathway plays a key role in the mechanism by which HDAC4 improves the survival rate and biofunction of chondrocytes.

Dual-branch hybrid network for lesion segmentation in gastric cancer images.

The effective segmentation of the lesion region in gastric cancer images can assist physicians in diagnosing and reducing the probability of misdiagnosis. The U-Net has been proven to provide segmentation results comparable to specialists in medical image segmentation because of its ability to extract high-level semantic information. However, it has limitations in obtaining global contextual information. On the other hand, the Transformer excels at modeling explicit long-range relations but cannot capture low-level detail information. Hence, this paper proposes a Dual-Branch Hybrid Network based on the fusion Transformer and U-Net to overcome both limitations. We propose the Deep Feature Aggregation Decoder (DFA) by aggregating only the in-depth features to obtain salient lesion features for both branches and reduce the complexity of the model. Besides, we design a Feature Fusion (FF) module utilizing the multi-modal fusion mechanisms to interact with independent features of various modalities and the linear Hadamard product to fuse the feature information extracted from both branches. Finally, the Transformer loss, the U-Net loss, and the fused loss are compared to the ground truth label for joint training. Experimental results show that our proposed method has an IOU of 81.3%, a Dice coefficient of 89.5%, and an Accuracy of 94.0%. These metrics demonstrate that our model outperforms the existing models in obtaining high-quality segmentation results, which has excellent potential for clinical analysis and diagnosis. The code and implementation details are available at Github, https://github.com/ZYY01/DBH-Net/ .

Chromosome-scale Genome Assembly of the Yellow Nutsedge (Cyperus esculentus).

The yellow nutsedge (Cyperus esculentus L. 1753) is an unconventional oil plant with oil-rich tubers, and a potential alternative for traditional oil crops. Here, we reported the first high-quality and chromosome-level genome assembly of the yellow nutsedge generated by combining PacBio HiFi long reads, Novaseq short reads, and Hi-C data. The final genome size is 225.6 Mb with an N50 of 4.3 Mb. More than 222.9 Mb scaffolds were anchored to 54 pseudochromosomes with a BUSCO score of 96.0%. We identified 76.5 Mb (33.9%) repetitive sequences across the genome. A total of 23,613 protein-coding genes were predicted in this genome, of which 22,847 (96.8%) were functionally annotated. A whole-genome duplication event was found after the divergence of Carex littledalei and Rhynchospora breviuscula, indicating the rich genetic resources of this species for adaptive evolution. Several significantly enriched GO terms were related to invasiveness of the yellow nutsedge, which may explain its plastic adaptability. In addition, several enriched Kyoto Encyclopedia of Genes and Genomes pathways and expanded gene families were closely related with substances in tubers, partially explaining the genomic basis of characteristics of this oil-rich tuber.

Whole-genome analysis revealed the growth-promoting mechanism of endophytic bacterial strain Q2H1 in potato plants.

Introduction: Endophytes are non-pathogenic inhabitants of healthy plant tissues and have been found to promote plant growth and health. The endophytic bacterial strain Q2H1 was isolated from the roots of the potato and was identified to exhibit growth-promoting effects in potato plants. Methods: Whole-genome sequencing was performed to reveal the mechanism underlying its growth-promoting effect. The obtained sequencing data of approximately 5.65 MB encompassed 5,533 coding sequences. Of note, nine secondary metabolite gene clusters, including siderophore gene clusters, closely associated with plant growth promotion (PGP) were predicted by antiSMASH software. Comparative genomic analysis revealed that Q2H1 belongs to the genus Peribacillus. By gene function annotation, those genes related to plant growth-promoting activities, including indole-3-acetic acid (IAA) synthesis in tryptophan metabolism, siderophore biosynthetic activity, phosphate solubilization, nitrogen fixation, and related genes, were summarized. IAA (14.4 µg/ml) was presumptively produced by Q2H1 using the Salkowski colorimetric method. A total of five genes, namely, phoU, pstB, pstA1, pstC, and pstS, were annotated for phosphate solubilization, which is associated with the ability of the Q2H1 strain to solubilize phosphate under in vitro conditions. Results: It is revealed that genes in the Q2H1 genome associated with nitrogen fixation belonged to three groups, namely, nitrogen fixation (nifU, sufU, salA, and nifS), nitrogen metabolism (nirA, nrtB, and nasA), and glutamate synthesis (glnA, gltB, gltD, and gudB), supported by evidence that Q2H1 grew on medium without nitrogen. We have also identified a siderophore gene cluster located on the chromosome of Q2H1, including seven genes (viz., rbsR, rhbf, rhbE, rhbD, rhbC, rhbA, ddc, and an unknown gene). In the in vitro assay, a prominent brown circle around the colony was produced on the chrome azurol S medium at 48 and 72 h post-inoculation, indicating that the siderophore gene cluster in Q2H1 harbored the ability to produce siderophores. Conclusion: In summary, these findings implied that identifying strain-specific genes for their metabolic pathways in bacterial endophytes may reveal a variety of significant functions of plant growth-promoting mechanisms.

Complete mitochondrial genome sequence and comparative analysis of the cultivated yellow nutsedge.

As a monocotyledonous plant in family Cyperaceae, yellow nutsedge (Cyperus esculentus L.) is unique in accumulating a substantial amount of oil in underground tubers and provides a model system for studying oil accumulation in nonseed tissues. However, no data on the mitochondrial and nuclear genome sequences of this species are available, which greatly limits our understanding of its evolutionary characteristics and some essential biological mechanisms. In the present study, we report the first complete mitochondrial genome sequence of the cultivated yellow nutsedge. The analysis of the genome showed that the yellow nutsedge mitochondrial genome is 1,002,696 bp in size and encodes 62 genes consisting of 36 protein-coding genes (PCGs), 20 transfer RNA (tRNA) genes, and six ribosomal RNA (rRNA) genes. Compared with other angiosperms, yellow nutsedge mitochondrial genome contains much higher percentage of noncoding sequences (95.36%). Sixteen plastid-derived fragments were identified to be strongly associated with mitochondrial genes including one intact plastid-related gene (ndhH). Comparative analysis with seven other sequenced plant mitochondrial genomes revealed that two syntenic gene clusters, rps3-rpl16 and rps12-nad3, are highly conserved in all plant mitochondrial genomes, and the mitochondrial genome of yellow nutsedge is more similar to those of monocotyledons in the gene order. Phylogenetic analysis based on 13 shared protein-encoding genes in eight plant species showed that yellow nutsedge is evolutionarily more closely related to monocotyledonary species. Overall, the species-specific features of the cultivated yellow nutsedge mitochondrial genome provide additional information for the evolutionary and comparative genomic studies in the yellow nutsedge and other Cyperus species of the Cyperaceae family.

Multienzyme System in Amorphous Metal-Organic Frameworks for Intracellular Lactate Detection.

Lactate is an important downstream product of glycolysis in living cells, and its level is highly related with diseases. On the basis of amorphous metal-organic frameworks (aMOFs), a multienzyme system consisting of lactate oxidase (LOx) and horseradish peroxidase (HRP) was established for intracellular lactate detection. By coencapsulation in aMOFs with proximity, LOx and HRP were delivered into cells, serving as artificially constructed organelles, exhibiting high activity and selectivity for the intracellular detection of the important metabolite lactate, which improved the signal to noise ratio by ∼650-fold. As demonstrated by both experimental and simulation results, the high efficiency was attributed to the short distance between the two types of enzymes coencapsulated in aMOFs. The concept of constructing multienzyme systems in this study shows promise for the detection of various intracellular metabolites.

Morpho-Agronomic and Biochemical Characterization of Accessions of Tiger Nut (Cyperus esculentus) Grown in the North Temperate Zone of China.

Tiger nut (Cyperus esculentus L.) has recently attracted increasing interest from scientific and technological communities because of its potential for serving as additional source of food, oil, and feed. The present study reports morphology and biochemical characterization of 42 tiger nut accessions collected from China and other counties performed in the 2020 and 2021 growing seasons at Nongan, Jilin Province. Assessment of variability of 14 agronomic traits including plant height, maturation, leaf width, tilling number, color, size, and shape: 100-tuber weight showed a wide range of phenotypic variation. The color, size, and shape and maturation of the tubers, as well as the leaf width, were the most distinct characteristics describing variation among the accessions. Compositional analyses of major nutritional components of the tubers reveals that this crop could be a source of high-value proteins, fatty acids, and carbohydrates. Specifically, tiger nut tubers contained high levels of starch, oil, and sugars, and significant amounts of fiber, Ca, P, and Na. Furthermore, the tubers appeared to be a good source of proteins as they contain 16 amino acids, including the essential ones. Amino acid profiles were dominated by aspartic acid followed by glutamic acid, leucine, alanine, and arginine. Overall, these results demonstrated that tiger nut is well adapted to the temperature and light conditions in the north temperate zone of China, even with a shorter growth season. The tiger nut accessions collected here exhibited wide variations for agronomical and biochemical traits, suggesting potential for potential for breeding improvement by maximizing the fresh tuber and grass yield based on the optimal selection of genetic characteristics in climate and soil conditions of northern China.

Enhanced resistance to soybean cyst nematode in transgenic soybean via host-induced silencing of vital Heterodera glycines genes.

Soybean cyst nematode (SCN, Heterodera glycines Ichinohe) is the most economically damaging pathogen affecting soybean production worldwide. Host-induced gene silencing provides a promising approach to confer resistance to plant parasitic nematodes. In the present study, we produced stable transgenic soybean plants individually harboring the inverted repeats of three essential H. glycines genes, Hg-rps23, Hg-snb1, and Hg-cpn1, and evaluated their resistance to SCN infection. Molecular characterization confirmed the stable integration of the hairpin double stranded (ds) RNA in host plants. Inoculation assays with SCN race 3 showed significant reduction of female index (FI, 11.84 ~ 17.47%) on the roots of T4 transgenic plants, with 73.29 ~ 81.90% reduction for the three RNA interference (RNAi) constructs, compared to non-transformed plants (NT, 65.43%). Enhanced resistance to SCN race 3 was further confirmed in subsequent generations (T5) of transgenic soybean. Moreover, when inoculated with SCN race 4 which was considered highly virulent to most of soybean germplasms and varieties, transgenic soybean plants also exhibited reduced FIs (9.96 ~ 23.67%) and increased resistance, relative to the NT plants (46.46%). Consistently, significant down-regulation in transcript levels of the Hg-rps23, Hg-snb1, Hg-cpn1 genes were observed in the nematodes feeding on the transgenic roots, suggesting a broad-spectrum resistance mediated by the host-mediated silencing of vital H. glycines genes. There were no significant differences in morphological traits between transgenic and NT soybean plants under conditions with negligible SCN infection. In summary, our results demonstrate the effectiveness of host-induced silencing of essential H. glycines genes to enhance broad-spectrum SCN resistance in stable transgenic soybean plants, without negative consequences on the agronomic performance.

Combining IOTA and Attribute-Based Encryption for Access Control in the Internet of Things.

Unauthorized resource access represents a typical security threat in the Internet of Things (IoT), while distributed ledger technologies (e.g., blockchain and IOTA) hold great promise to address this threat. Although blockchain-based IoT access control schemes have been the most popular ones, they suffer from several significant limitations, such as high monetary cost and low throughput of processing access requests. To overcome these limitations, this paper proposes a novel IoT access control scheme by combining the fee-less IOTA technology and the Ciphertext-Policy Attribute-Based Encryption (CP-ABE) technology. To control the access to a resource, a token, which records access permissions to this resource, is encrypted by the CP-ABE technology and uploaded to the IOTA Tangle (i.e., the underlying database of IOTA). Any user can fetch the encrypted token from the Tangle, while only those who can decrypt this token are authorized to access the resource. In this way, the proposed scheme enables not only distributed, fee-less and scalable access control thanks to the IOTA but also fine-grained attribute-based access control thanks to the CP-ABE. We show the feasibility of our scheme by implementing a proof-of-concept prototype system using smart phones (Google Pixel 3XL) and a commercial IoT gateway (NEC EGW001). We also evaluate the performance of the proposed scheme in terms of access request processing throughput. The experimental results show that our scheme enables object owners to authorize access rights to a large number of subjects in a much (about 5 times) shorter time than the existing access control scheme called Decentralized Capability-based Access Control framework using IOTA (DCACI), significantly improving the access request processing throughput.

Efficient identification of genomic insertions and flanking regions through whole-genome sequencing in three transgenic soybean events.

Genomic insertions and flanking regions of transgenes in host genomes constitute a critical component of precise molecular characterization and event-specific detection, which are required in the development and assessment for regulatory approval of genetically modified (GM) crops. Previously, we reported three transgenic soybean events harboring the inverted repeats of the soybean mosaic virus NIb (nuclear inclusion b) gene, exhibiting significantly enhanced resistance to multiple Potyvirus strains. To facilitate safety assessment and event-specific detection, we identified the transgene insertion sites and flanking sequences of the events L120, L122, and L123 using whole-genome sequencing. More than 14.48 Gb sequence data (13 × coverage) were generated using the Illumina HiSeq Xten platform for each event. The sequence reads corresponding to boundaries of inserted T-DNA, and associated native flanking sequences were identified by bioinformatic comparison with the soybean reference genome (Wm82.a2.v1) and the transformation vector sequence. The results indicated that two T-DNA insertions occurred in L120, on Chr07 and Chr13, while L122 and L123 showed single insertions, on Chr02 and Chr06, respectively. Based on the flanking sequences of the inserted T-DNA, the event-specific detection for each event was established using specific PCR primers, and PCR amplification followed by sequencing of PCR products further confirmed the putative insertion loci and flanking regions in the transgenic lines. Our results demonstrate the efficacy and robustness of whole-genome sequencing in identifying the genomic insertions and flanking regions in GM crops. Moreover, the characterization of insertion loci and the establishment of event-specific detection will facilitate the application and development of broad-spectrum virus-resistant transgenic soybean cultivars.

Enhanced tolerance to Phytophthora root and stem rot by over-expression of the plant antimicrobial peptide CaAMP1 gene in soybean.

BACKGROUND: Antimicrobial peptides play important roles in both plant and animal defense systems. Moreover, over-expression of CaAMP1 (Capsicum annuum antimicrobial protein 1), an antimicrobial protein gene isolated from C. annuum leaves infected with Xanthomonas campestris pv. vesicatoria, confers broad-spectrum resistance to hemibiotrophic bacterial and necrotrophic fungal pathogens in Arabidopsis. Phytophthora root and stem rot (PRR), caused by the fungus Phytophthora sojae, is one of the most devastating diseases affecting soybean (Glycine max) production worldwide. RESULTS: In this study, CaAMP1 was transformed into soybean by Agrobacterium-mediated genetic transformation. Integration of the foreign gene in the genome of transgenic soybean plants and its expression at the translation level were verified by Southern and western blot analyses, respectively. CaAMP1 over-expression (CaAMP1-OX) lines inoculated with P. sojae race 1 exhibited enhanced and stable PRR tolerance through T2-T4 generations compared with the wild-type Williams 82 plants. Gene expression analyses in the transgenic plants revealed that the expression of salicylic acid-dependent, jasmonic acid-dependent, and plant disease resistance genes (R-genes) were significantly up-regulated after P. sojae inoculation. CONCLUSIONS: These results indicate that CaAMP1 over-expression can significantly enhance PRR tolerance in soybean by eliciting resistance responses mediated by multiple defense signaling pathways. This provides an alternative approach for developing soybean varieties with improved tolerance against soil-borne pathogenic PRR.

Distribution, sources, risks, and vitro DNA oxidative damage of PM2.5-bound atmospheric polycyclic aromatic hydrocarbons in Urumqi, NW China.

Research has focused on the impacts of polycyclic aromatic hydrocarbons (PAHs) in the atmosphere due to their potential carcinogenicity. In this study, we investigated the seasonal variation, sources, incremental lifetime cancer risks (ILCRS), and vitro DNA oxidative damage of PAHs in Urumqi in NW China. A total of 72 atmospheric samples from Urumqi were collected over a year (September 2017-September 2018) and were analyzed for 16 PAHs that are specifically prioritized by the U.S Environmental Protection Agency (U·S EPA). The highest PAHs concentrations were in winter (1032.66 ng m-3) and lowest in spring (146.00 ng m-3). Middle molecular weight PAHs with four rings were the most abundant species (45.28-61.19% of the total). The results of the diagnostic ratio and positive matrix factorization inferred that the major sources of atmospheric PAHs in Urumqi were biomass burning, coking, and petrogenic sources (52.9%), traffic (30.1%), coal combustion (8.9%), and the plastics recycling industry (8.1%). ILCRS assessment and Monte Carlo simulations suggested that for all age groups PAHs cancer risks were mainly associated with ingestion and dermal contact and inhalation was negligible. The plasmid scission assay results showed a positive dose-response relationship between PAHs concentrations and DNA damage rates, demonstrating that toxic PAHs was the primary cause for PM2.5-induced DNA damage in the air of Urumqi.

Exploiting Smart Contracts for Capability-Based Access Control in the Internet of Things.

Due to the rapid penetration of the Internet of Things (IoT) into human life, illegal access to IoT resources (e.g., data and actuators) has greatly threatened our safety. Access control, which specifies who (i.e., subjects) can access what resources (i.e., objects) under what conditions, has been recognized as an effective solution to address this issue. To cope with the distributed and trust-less nature of IoT systems, we propose a decentralized and trustworthy Capability-Based Access Control (CapBAC) scheme by using the Ethereum smart contract technology. In this scheme, a smart contract is created for each object to store and manage the capability tokens (i.e., data structures recording granted access rights) assigned to the related subjects, and also to verify the ownership and validity of the tokens for access control. Different from previous schemes which manage the tokens in units of subjects, i.e., one token per subject, our scheme manages the tokens in units of access rights or actions, i.e., one token per action. Such novel management achieves more fine-grained and flexible capability delegation and also ensures the consistency between the delegation information and the information stored in the tokens. We implemented the proposed CapBAC scheme in a locally constructed Ethereum blockchain network to demonstrate its feasibility. In addition, we measured the monetary cost of our scheme in terms of gas consumption to compare our scheme with the existing Blockchain-Enabled Decentralized Capability-Based Access Control (BlendCAC) scheme proposed by other researchers. The experimental results show that the proposed scheme outperforms the BlendCAC scheme in terms of the flexibility, granularity, and consistency of capability delegation at almost the same monetary cost.

Overexpression of the chitinase gene CmCH1 from Coniothyrium minitans renders enhanced resistance to Sclerotinia sclerotiorum in soybean.

Pathogenic fungi represent one of the major biotic stresses for soybean production across the world. Sclerotinia sclerotiorum, the causal agent of Sclerotinia stem rot, is a devastating fungal pathogen that is responsible for significant yield losses in soybean. In this study, the chitinase gene CmCH1, from the mycoparasitic fungus Coniothyrium minitans, which infects a range of ascomycetous sclerotia, including S. sclerotiorum and S. minor, was introduced into soybean. Transgenic plants expressing CmCH1 showed higher resistance to S. sclerotiorum infection, with significantly reduced lesion sizes in both detached stem and leaf assays, compared to the non-transformed control. Increased hydrogen peroxide content and activities of defense-responsive enzymes, such as peroxidase, superoxide dismutase, phenylalanine ammonia lyase, and polyphenoloxidase were also observed at the infection sites in the transgenic plants inoculated with S. sclerotiorum. Consistent with the role of chitinases in inducing downstream defense responses by the release of elicitors, several defense-related genes, such as GmNPR2, GmSGT-1, GmRAR1, GmPR1, GmPR3, GmPR12, GmPAL, GmAOS, GmPPO, were also significantly upregulated in the CmCH1-expressing soybean after inoculation. Collectively, our results demonstrate that overexpression of CmCH1 led to increased accumulation of H2O2 and up-regulation of defense-related genes and enzymes, and thus enhanced resistance to S. sclerotiorum infection while showing no detrimental effects on growth and development of soybean plants.

Packaging and delivering enzymes by amorphous metal-organic frameworks.

Enzymatic catalysis in living cells enables the in-situ detection of cellular metabolites in single cells, which could contribute to early diagnosis of diseases. In this study, enzyme is packaged in amorphous metal-organic frameworks (MOFs) via a one-pot co-precipitation process under ambient conditions, exhibiting 5-20 times higher apparent activity than when the enzyme is encapsulated in corresponding crystalline MOFs. Molecular simulation and cryo-electron tomography (Cryo-ET) combined with other techniques demonstrate that the mesopores generated in this disordered and fuzzy structure endow the packaged enzyme with high enzyme activity. The highly active glucose oxidase delivered by the amorphous MOF nanoparticles allows the noninvasive and facile measurement of glucose in single living cells, which can be used to distinguish between cancerous and normal cells.

FvC5SD overexpression enhances drought tolerance in soybean by reactive oxygen species scavenging and modulating stress-responsive gene expression.

KEY MESSAGE: Overexpression of FvC5SD improves drought tolerance in soybean. Drought stress is one of the most important abiotic stress factors that influence soybean crop quality and yield. Therefore, the creation of drought-tolerant soybean germplasm resources through genetic engineering technology is effective in alleviating drought stress. FvC5SD is a type of C-5 sterol desaturase gene that is obtained from the edible fungus Flammulina velutipes. This gene has good tolerance to the effects of stresses, including drought and low temperature, in yeast cells and tomato. In this study, we introduced the FvC5SD gene into the soybean variety Shennong9 through the Agrobacterium-mediated transformation of soybean to identify drought-tolerant transgenic soybean varieties. PCR, RT-PCR, and Southern blot analysis results showed that T-DNA was inserted into the soybean genome and stably inherited by the progeny. The ectopic expression of FvC5SD under the control of a CaMV 35S promoter in transgenic soybean plants enhanced the plant's tolerance to dehydration and drought. Under drought conditions, the transgenic plants accumulated lower levels of reactive oxygen species and exhibited higher activities and expression levels of enzymes and cell than wild-type soybean. iTRAQ analysis of the comparative proteomics showed that some exogenous genes coding either functional or regulatory proteins were induced in the transgenic lines under drought stress. FvC5SD overexpression can serve as a direct and efficient target in improving drought tolerance in soybean and may be an important biotechnological strategy for trait improvement in soybean and other crops.