Cystathionine γ-lyase (CGL) mutant cyst(e)inase effectively degrades cystine (CSSC) and cysteine (L-Cys), exhibiting antitumor activity through redox homeostasis disruption in tumor cells. However, its efficacy has not yet met the requirements for clinical application. To enhance the catalytic activity and thermodynamic stability of cyst(e)inase for CSSC and L-Cys degradation, this study focused on evolutionarily conserved amino acid residues using virtual site-saturation mutagenesis coupled with free energy calculations to identify potential beneficial mutants. The screening of 119 amino acid sites identified 32 promising mutants through free energy analysis. Experimental validation revealed that 8 mutants exhibited enhanced activity toward CSSC and L-Cys. Among these, the top-performing mutant, S206I, demonstrated 40% and 60% higher activity against CSSC and L-Cys, respectively, accompanied by 23% and 230% k_cat/K_m improvements. Additionally, its thermal stability (T_m) was elevated by 1.89 ℃. This study employed an efficient and straightforward design strategy, produced enhanced mutants with enhanced activity and stability, establishing a framework for subsequent therapeutic enzyme engineering.

To determine the structure of BetA fromPseudomonas denitrificans, the recombinant plasmid pET-28a-PdBetA was constructed to express and purifyPdBetA protein for crystallization experiments. X-ray diffraction analysis of the protein crystals yielded a 1.8  resolution structure ofPdBetA. ThePdBetA structure consists of a β sandwich fold comprising an antiparallel three-stranded β-sheet together with a parallel five-stranded β-sheet, and an antiparallel six-stranded β-sheet with two approximately parallel α-helices on the same side. Structural comparison and sequence alignment with its homologs suggested a potential flavin adenine dinucleotide binding pocket inPdBetA.

In this study, the microbial strain designated as ZH-5 and capable of producting laccase was isolated from soil by plate chromogenic method and identified asSerratia marcescensby 16S rDNA and physiological and biochemical characteristics. Genome sequencing results showed that the genome size of strain ZH-5 was 5055904 bp, with GC content accounted for 59.55%, 4643 genes, and a total coding length of 4423485 bp, accounting for 87.49% of the genome. The genome contained a laccase gene (GE001633) and several heterologous biomass-degradation pathways. The enzyme reaction system was optimized firstly. The medium components of carbon and nitrogen sources were optimized further through single-factor optimization and orthogonal experiments. Based on single-factor and enzyme-catalyzed reaction system optimization, the medium composition was optimized through orthogonal experiments combining carbon and nitrogen sources. After 6 days of cultivation, the ZH-5 strain produced laccase activity of (24.68±0.76) U/mL in modified LB liquid medium containing 1 mmol/L Cu2+, 20 g/L peptone, 10 g/L yeast powder, and 20 g/L glycerol, representing a 34.74-fold increase compared to pre-optimization levels. In addition, the growth characteristics of ZH-5 were measured. The results showed that ZH-5 had better acid and salt tolerance and a broader temperature range for growth. Through the whole genome sequencing and gene function analysis of ZH-5, the activity of laccase was fully characterized, providing a theoretical basis for the subsequent application research based on ZH-5 laccase.

In this study, coevolutionary residue analysis was used to identify key functional sites affecting the robustness of C57E10 (derived from Nit6803), an enzyme that efficiently catalyzes the production of nicotinic acid from 3-cyanopyridine. Saturation combinatorial mutations were designed directly on the coevolutionary residues and the potential thermostability-enhanced mutations were evaluated by the change of folding free energy (ddG). Two mutants, K44Y and Q120L/A166L, were screened out, which maintained high catalytic activity with significantly enhanced thermostability, and the half-life in C57E10 at 50 ℃ was increased from 15.1 min to 34.5 min and 21.4 min, respectively. However, the further combination of K44Y and Q120L/A166L resulted in a dramatic decrease in thermostability, revealing strong negative epistasis. Structural analysis showed that K44Y enhanced its hydrophobic interaction with I274, whereas Q120L/A166L established denser hydrophobic interaction networks, thereby improving the structural stability. In this study, the nitrilase with high thermostability was obtained, and the strategy based on combination of coevolutionary residue analysis and mutation calculations also provided an applicable reference for the rational design of other enzymes.

This study investigated the effects and potential mechanisms of Lcc5, a fungal laccase derived fromCoprinopsis cinerea, on the quality and physicochemical properties of rye bread. Rye dough was treated with varying concentrations of Lcc5, and comprehensive analyses were performed to assess the structural and textural attributes of both dough and bread. Analytical methods included rheological measurements, texture profile analysis (TPA), free sulfhydryl group determination, and scanning electron microscopy (SEM) imaging. From the texture and rheological results, the addition of Lcc5 increased the elastic (G′) and viscous modulus (G″) of rye dough and affected the gluten strength of the dough, rye bread quality was improved. The hardness, chewiness and stickiness of rye bread were significantly reduced when Lcc5 was added up to 6 U. Microstructural observations using SEM indicated that Lcc5 promoted protein cross-linking within the gluten network. This cross-linking led to a smoother and tighter dough surface, with a significant decrease in the exposure of starch granules. These findings suggested that Lcc5 effectively improved the textural and structural qualities of rye bread by enhancing the degree of gluten network cross-linking and, consequently, gluten strength. Lcc5 served as a potent improver for rye bread and dough, offering a method to optimize the sensory and structural properties of rye-based products. This study provided a theoretical basis for the optimization of rye bread and dough processing.

Utilizing ultra-high performance liquid chromatography-quadrupole-electrostatic field orbitrap high resolution mass spectrometry (UPLC-Q-Exactive Orbitrap-MS/MS), a total of 68 chemical constituents were identified from sea buckthorn fruit. Network pharmacology analysis and molecular docking predicted 8 potential antibacterial compounds in sea buckthorn fruit, including corilagin, epigallocatechin, and gallocatechin. These compounds might exhibit favorable binding affinities with key targets including TNF, IL6, and AKT1, potentially exerting antibacterial effects via the modulation of signaling pathways including PI3K-Akt and IL-17. Furthermore,in vitrominimum inhibitory concentration (MIC) assays confirmed the antibacterial efficacy of corilagin, epigallocatechin, gallocatechin, and taxifolin. Notably, the combination of corilagin and epigallocatechin demonstrated a synergistic inhibition againstStaphylococcus aureus, reducing MICs to 31.25 μg/mL and 62.5 μg/mL, respectively. This research, leveraging high-resolution mass spectrometry technology and integrating data from multiple databases, comprehensively characterized the chemical profile of sea buckthorn fruit, establishing a solid foundation for understanding its pharmacological properties and traditional medicinal efficacy. By integrated network pharmacology, molecular docking techniques, andin vitroexperiments, this study represented the first discovery of multiple antibacterial chemical components in sea buckthorn fruit, revealing their potential mechanisms, providing a theoretical basis for the application of sea buckthorn fruit in antibacterial and anti-infective therapies.

This study focused on the impact of tidal changes on the microbial communities in the rhizosphere and leaves ofZostera japonica. Nanopore amplicon sequencing was used to analyze the diversity, structure, and functional characteristics of the microbial communities of the leaves and rhizospheres across the high tide zone, low tide zone, and subtidal area. Environmental factors were also evaluated to assess their influence on the microbial communities. The results showed that the microbial abundance and diversity of the leaves and rhizosphere was similar to tidal change trends, peaking in low-tide zones and reaching minima in subtidal areas. Tidal variation significantly altered the α-diversity index of the leaf microbiota （P<0.05）, without affecting the rhizosphere microbiota （P>0.05）. In the subtidal and high tide zones, the leaf and rhizosphere microbial communities exhibited a higher abundance of characteristic taxa. In the high tide zone, leaf microbiota adapted to the exposed environment and were involved in sulfur cycling, while in the low tide zone, leaf microbiota showed strong organic matter degradation capacity. In the subtidal zone, leaf microbes not only efficiently degradated organic matter but also participated in nitrogen and sulfur cycling. High-tide rhizosphere microbes prioritized sulfur cycling and redox homeostasis, contrasting with low-tight populations specializing in nitrogen fixation and degradation. The subtidal rhizosphere were adapted to low-oxygen environments and participated in sulfur cycling. Sediment total organic carbon （TOC） significantly affected the composition and distribution of rhizosphere communities. From the high tide zone to the subtidal zone, the rhizosphere microbial ability to utilize organic matter, sulfur compounds, and nitrogen respiratory activity increased, while aerobic heterotrophic activity decreased. Leaf microbiota displayed enhancing nitrate reduction but reducing sulfur respiration and photoautotrophy. These findings provide theoretical support for the conservation strategies ofZ. japonica.

In this study, theLOXgene family inNelumbo nuciferawas identified and analyzed, and its expression patterns in various tissues were quantified using RT-qPCR. The results revealed that the identified 9LOXgene family members in theN. nuciferagenome belong to the 9-LOX and 13-LOX subfamilies. Subcellular localization predicted that the 9 proteins were primarily located in the cytoplasm or chloroplast. Secondary and tertiary structure predictions of the proteins showed that LOX proteins were predominantly composed of α-helices and random coils, with most being acidic proteins. Gene structure and conserved motif analysis revealed divergence among evolutionary branches but conservation within subfamilies. The promoter regions of theNnLOXgenes were rich in stress response, light response, and hormone response elements. Expression pattern analysis revealed thatNnLOX1exhibited a higher expression level in the stamens,NnLOX2was specifically expressed in the lotus seeds,NnLOX4showed significant expression in the petals, andNnLOX8andNnLOX9were primarily expressed in the lotus rhizomes. These findings suggested that the expression of lotusLOXgenes was tissue-specific, which provided a theoretical basis for further clarifying the function of theN. nucifera LOXgene and its role in plant physiological processes.

This study aimed to investigate the mechanism by whichPolygonatum sibiricumpolysaccharides (PSP) modulate depression-like behaviors via the Netrin-1/DCC pathway in the medial prefrontal cortex (mPFC). After establishing the learned helplessness (LH) and chronic unpredictable mild stress (CUMS) depression models, PSP (400 mg/kg) was administered continuously for 14 days. The depressive-like behaviors of the mice were evaluated using the open field test, tail suspension test, and forced swimming test. Western blot analysis was performed to detect the Netrin-1/DCC signaling pathway in the medial prefrontal cortex of mice, as well as the expression levels of GFAP, NF-κB, IL-1β, GluN2A, GluN2B, and PSD95. The results indicated that, compared to the control group, the model group of mice exhibited increased immobility time in both the tail suspension and forced swimming tests. Administration of PSP reduced depression-like behaviors in model mice. Concurrently, in the mPFC of the model group mice, the expression levels of Netrin-1/DCC, GFAP, NF-κB, and IL-1β, which were associated with inflammatory responses, as well as GluN2A and GluN2B related to synaptic function, were elevated, while the expression level of PSD95 was reduced. PSP was found to inhibit the activation of Netrin-1/DCC, downregulate GFAP, NF-κB, IL-1β, GluN2A, and GluN2B, and upregulate PSD95 expression. The study indicated that the Netrin-1/DCC signaling pathway was abnormally activated during the onset of depression, and that PSP might exert its antidepressant effects by inhibiting this pathway, thereby regulating synaptic function and inflammatory responses. This process might be a crucial mechanism through which PSP improves depressive symptoms.

The research team previously utilized miRNA sequencing technology to identify a differentially expressed and highly conserved miRNA, miR-196b-5p, in the soleus and longissimus dorsi muscles of large white pigs. Previous studies have demonstrated that miR-196b-5p promotes the proliferation and differentiation of C2C12 myoblasts, its role in myofiber type conversion remains unclear. To explore the effects and regulatory mechanisms of miR-196b-5p on skeletal muscle fiber type conversion, this study used mice as experimental subjects and systematically analyzed the regulatory role of miR-196b-5p by injecting AAV-miR-196-5p and AAV-empty into mice muscles. The statistical results showed that AAV-miR-196-5p administration did not significantly influence the body weight, feed intake, or tissue weight of the mice （P>0.05）, confirming no interference with normal growth. H&E staining results revealed that miR-196b-5p overexpression significantly enlarged fiber cross-sectional areas in both tibialis anterior muscle （TA） and gastrocnemius muscle （GAS） （P<0.05）. Real-time fluorescence quantitative PCR results demonstrated that miR-196b-5p overexpression downregulated the slow-twitch muscle fiber marker genes,MyHC ⅠandMyHC Ⅱa（P<0.05） but upregulated the fast-twitch muscle fiber marker gene,MyHC Ⅱb（P<0.05）. Additionally, the overexpression of miR-196b-5p significantly reduced the mRNA expression levels of mitochondrial oxidative phosphorylation related genes （COX5B,COX6C,COX7A1, andCOX8B） （P<0.01）, suggesting that miR-196b-5p may inhibit the oxidative metabolism of skeletal muscle. KEGG enrichment analysis of the target genes of miR-196b-5p revealed that these genes were mainly enriched in the p38 mitogen-activated protein kinase （p38-MAPK） pathway and the AMP-activated protein kinase （AMPK） pathway. Western blot showed that the overexpression of miR-196b-5p significantly reduced the protein phosphorylation level of p38-MAPK （P<0.05） while significantly increasing the protein phosphorylation level of AMPK （P<0.05）. Furthermore, overexpression of miR-196b-5p significantly reduced the protein expression level of the slow-twitch muscle fiber marker gene MyHCⅠ （P<0.05）. These findings suggested that miR-196b-5p played a crucial role in skeletal muscle fiber type conversion via AMPK activation and p38-MAPK inhibition, thereby suppressing oxidative metabolism, reducing the formation of slow-twitch muscle fibers, and promoting the formation of fast-twitch muscle fibers.

In the present study, an optimized maximum entropy model （MaxEnt） was used to predict both the potential habitat distribution and its dynamics under future climate scenarios across different greenhouse gas emission modes for the genusEothenomys. The findings indicated that the geographic distribution ofEothenomysprimarily encompasses regions within Yunnan, Sichuan, Chongqing, Guizhou, Guangxi, Guangdong, Fujian, and southeastern Xizang in China. Moreover, areas exhibiting high suitability forEothenomyshabitat were predominantly clustered in the northern Yunnan, southern Sichuan, northwestern Guizhou and southeastern Xizang regions. Warmest-quarter precipitation was the key climatic variable affecting the distribution ofEothenomys. The distribution ofEothenomysshowed irregular patterns across different greenhouse gas emission patterns. Although the total area of predicted distribution zones ofEothenomysin the 2050s, 2070s, and 2090s showed minimal changes, high-suitability habitats decreased accompanied by slight shifts in distribution centers, primarily concentrated in the eastern region at the border of Yunnan and Sichuan. In the future, someEothenomyspopulations might migrate to higher altitudes, while others could expand into eastern China and coastal areas. The distribution areas ofEothenomysexhibited significant altitude differences and severe habitat fragmentation, highlighting the importance of enhanced monitoring and protection of the species to preserve local ecological diversity, and the need of implementing measures to prevent rodent damage.

The regulation of endothelial cell morphological changes and pathophysiological functions, including proliferation, migration, senescence, apoptosis, secretion and phenotypic switch, is dependent on blood flow derived shear stress. As a monolayer lining the vascular lumen, endothelial cells’ surface harbors diverse mechanoreceptors and mechanosensitive complexes that respond to different shear stress patterns, subsequently regulating gene expression and cellular behavior via mechanotransduction pathways. In particular, laminar shear stress （LSS） induces the orderly arrangement of endothelial cells, an increase in the levels of vascular protection-related genes, and the maintenance of cellular homeostasis. Conversely, low oscillatory shear stress （OSS, also known as disturbed flow） disrupts endothelial alignment while activating inflammatory and oxidative stress pathways, ultimately inducing endothelial dysfunction. This article comprehensively reviewed the latest advances on blood flow derived shear stress-regulated endothelial cell function, providing mechanistic insights into biomechanical regulation of vascular function.

This study presents an overview of the types and quantities of functional components in Antarctic krill oil and summarizes the skin-care effects and mechanisms of each component. The ingredients of Antarctic krill oil primarily comprise phospholipids, astaxanthin, tocopherols, and polyunsaturated fatty acids. These components play significant roles in alleviating skin inflammation, combating photoaging, assisting psoriasis treatment, moisturizing, and promoting wound healing, thus conferring moisturizing and skin-repairing benefits. Through comprehensive analysis of animal studies and some clinical trials, the skin care mechanism of Antarctic krill oil are elucidated through anti-inflammatory, antioxidant, gene expression regulation, cellular repair, and wound healing pathways. Finally, the potential of Antarctic krill oil in protecting the skin is pointed out and the limitations existing in current research are analyzed, establishing a foundation for future applications of Antarctic krill oil in skin-care.

n this study, an RPA-CRISPR/Cas12a-LFS assay was developed for the detection of white spot syndrome virus （WSSV） based on recombinase polymerase amplification （RPA） combined with CRISPR/Cas12a system and lateral flow strips （LFS）, which was rapid, sensitive and simple to operate. The lower limit of the detection method for RPA-CRISPR/Cas12a-LFS was 1.12×101copies of vitiligo syndrome virus DNA per reaction, and the whole process could be completed within 1 h. Furthermore, the assay showed no cross-reactivity with other common shrimp viruses or healthy shrimp genomes. When testing 110 clinical shrimp samples collected from shrimp farms, the results of this study were completely consistent with the results of real-time polymerase chain reaction （PCR） assays. In conclusion, the detection method （RPA-CRISPR/Cas12a-LFS） constructed in this study shows great potential as a novel detection scheme for the early diagnosis of WSSV in shrimp.

A novel detection method for hepatitis A virus (HAV) was developed to meet the rapid detection needs, based on reverse transcription-recombinase polymerase amplification (RT-RPA) technology combined with lateral flow strips (LFS). Genotype Ⅰ-Ⅲ sequences of HAV were downloaded from the GenBank nucleotide database, and genetic typing and analysis were conducted to design primers and probes of RT-RPA. This method could complete HAV detection within 20 minutes at 42 ℃, with a sensitivity of 10 copies/μL, exhibiting high specificity, no cross-reactivity with other enteric viruses, and a repeatability of 71.4%. Compared to traditional RT-qPCR technology, this method demonstrated greater sensitivity and easier operation. The establishment of the RT-RPA-LFS detection technology provided technical support for on-site rapid diagnosis of HAV, which was of great significance for improving the efficiency of HAV detection and preventing and controlling foodborne outbreaks caused by HAV. With widespread HAV vaccine use, the development of this detection technology has potential applications for distinguishing vaccine strains from wild strains, as well as meeting the future needs for multiplex virus detection, providing new technical approaches for food safety monitoring and public health fields.

To promote the development of synthetic biology and the cultivation of high-level talents, the curriculum of synthetic biology for the professional degree graduates majoring in biotechnology and engineering in the field of biology and medicine is offered by Hunan Agricultural University. Firstly, the curriculum objectives of synthetic biology were introduced. Secondly, according to the characteristics of students, the teaching content and teaching arrangements of synthetic biology curriculum were reasonably designed to stimulate students’ learning interest and enhance learning outcomes. Finally, the curriculum content was taught via the “offline + online” mode, including the thematic discussion and case teaching, and the learning outcomes were assessed through teaching questionnaires and course performance evaluations. Under the background of the new quality productive forces, the construction and exploration of synthetic biology curriculum for the professional degree graduate in the field of biology and medicine were proposed to enhance students’ scientific research and innovation capabilities and innovation ability and to support the development of high-quality talents.

Addressing the significant imbalance in national virtual simulation experimental teaching resource distribution, particularly in Western regions where quality virtual resources are scarce, this study demonstrated the research and outcomes of a collaboration between Nanjing Agricultural University and Xizang Agricultural and Animal Husbandry University in the cross-regional co-construction and sharing of biological virtual simulation teaching resources, as well as the collaborative innovation of teaching models from 2017. By integrating existing virtual simulation teaching resources, the universities co-developed virtual simulation teaching resources with regional characteristics. Through establishing virtual teaching and research rooms, two universities explored and practiced collaborative innovation in teaching modes of virtual simulation experiment. The related work vigorously enhanced the information technology level of Western universities, assisted in curriculum construction and teaching reform, and provided a feasible path for the co-construction and sharing of virtual simulation experimental teaching resources for universities in Eastern and Western China.

Considering the abundance ofDrosophilaspecies and their ease of collection/culture, a serial experiment on genetic diversity was designed usingDrosophilaflies as experimental materials. These experiments integrated and extended the established experimental system. The experimental series comprised 5 components: observation, collection, and culture ofDrosophila, species identification via DNA barcoding, creation and observation of salivary gland chromosome specimens, isozyme analysis, and SSR analysis. These experiments collectively assessed the genetic diversity ofDrosophilaat the phenotypical, chromosome, protein, and DNA levels. This multi-tiered approach featured progressive and exploratory characteristics. Using consistent experimental samples throughout the series enables students to systematically understand the knowledge and technology of genetics and comprehensively improve their experiment capabilities.

The bioengineering major of Hefei University has seized the opportunity of “double first-class” construction, assisted students to form scientific innovation teams under the collaborative guidance from academic mentors, science and innovations mentors, and enterprise mentors. The three mentor groups collaboratively established an ecosystem for innovation and entrepreneurship, which was beneficial for teaching, learning, competition, research and innovation. Specifically, students from different grades and majors were formed “interdisciplinary and cross-grade” scientific innovation teams by the integrated traditional classroom instruction with a variety of other educational scenarios, and their internal motivation for learning through interest were activated, while igniting their external drive through academic competitions. By assigning enterprise projects to scientific innovation teams, the transformation of theoretical knowledge into projects, the conversion of innovation achievements into practical products, and the development of students into innovative talents were accelerated. Consequently, the students’ awareness and capabilities in innovation and entrepreneurship were raised and strengthened, and the commercialization of their innovative achievements was promoted. In sum, through the construction and practice of the innovation and entrepreneurship ecosystem, the bioengineering national first-class undergraduate major construction was continuously developed and aligned the cultivation of innovation and entrepreneurship talents with the needs from regional industries, and thereby comprehensively enhanced the ability to cultivate professionals for serving the high-quality development of the regional economy.

In the future, engineering education will be transnational and enhance industrial engagement. Relying on first-class majors, the curriculum team has comprehensively arranged the breadth and depth of curriculum bilingual construction to improve talent development efficiency. A competency-based bilingual teaching model has been constructed. The completion of curriculum training goals is ensured through the multi-dimensional simultaneous development of quality training, teaching design, feature highlighting, practical skills and information integration. In this study, by curating instructional materials and strengthening faculty allocation, bilingual teaching resources have been improved and enriched, ensuring alignment with international disciplinary standards. By establishing tiered English proficiency benchmarks tailored to individual student’ s needs, designing discipline-specific English discussions on regional economic revitalization, integrating online and offline full-scene contexts, and optimizing assessment systems, this study implemented multidimensional teaching strategies to fully integrate bilingual education into the bioseparation engineering curriculum. Course evaluations indicated that students’  practical application of professional English has been strengthened. This study provides a new teaching paradigm for improving the quality of bioengineering talent training and enhancing the international influence and competitiveness of the “new engineering” of bioengineering.