Precise Molecular Subtyping Reveals Heterogeneity of Lung Adenocarcinoma Based on DNA Methylation.

BACKGROUND: Due to the high heterogeneity of lung adenocarcinoma (LUAD), which restricts the effectiveness of therapy, precise molecular subgrouping of LUAD is of great significance. Clinical research has demonstrated the significant potential of DNA methylation as a classification indicator for human malignancies. METHODS: WGML framework (which was developed based on weighted gene correlation network analysis (WGCNA), Gene Ontology (GO), and machine learning) was developed to precisely subgroup molecular subtypes of LUAD. This framework included two parts: the WG algorithm and the machine learning part. The WG algorithm part was an original algorithm used to obtain a crucial module, which was characterized by weighted correlation network analysis, functional annotation, and mathematical algorithms. The machine learning part utilized the Boruta algorithm, random forest algorithm, and Gradient Boosting Regression Tree algorithm to select feature genes. Then, based on the results of the WGML framework, subtypes were computed by the hierarchical clustering algorithm. A series of analyses, including dimensionality reduction methods, survival analysis, clinical stage analysis, immune infiltration analysis, tumor environment analysis, immune checkpoints analysis, TIDE analysis, CYT analysis, somatic mutation analysis, and drug sensitivity analysis, were utilized to demonstrate the effectiveness of subgrouping. GEO datasets were used to externally validate the results. Meanwhile, another subgrouping method of LUAD from another study was employed to compare with the WGML framework. RESULT: By importing DNA methylation data into the WGML framework, nine genes were obtained to further subgroup LUAD. Three subtypes, the Carcinogenesis subtype, Immune-infiltration subtype, and Chemoresistance subtype, were identified. The dimensionality reduction method exhibited great distinctness between subtypes. A series of analyses were employed to exhibit the difference among the three subtypes and to demonstrate the accuracy of the definition of subtypes. Besides, the WGML framework was compared with a LUAD subgrouping method from another research, which demonstrated that WGML had better efficiency for subgrouping LUAD. CONCLUSION: This study provides a novel LUAD subgrouping framework named WGML for the accurate subgrouping of lung adenocarcinoma.

Foxo1 is an iron-responsive transcriptional factor regulating systemic iron homeostasis.

The liver plays a crucial role in maintaining systemic iron homeostasis by secreting hepcidin, which is essential for coordinating iron levels in the body. Imbalances in iron homeostasis are associated with various clinical disorders related to iron deficiency or iron overload. Despite the clinical significance, the mechanisms underlying how hepatocytes sense extracellular iron levels to regulate hepcidin synthesis and iron storage are not fully understood. In this study, we identified Foxo1, a well-known regulator of macronutrient metabolism, that translocates to the nucleus of hepatocytes in response to high-iron feeding, holo-transferrin, and BMP6 treatment. Furthermore, Foxo1 plays a crucial role in mediating hepcidin induction in response to both iron and BMP signals by directly interacting with evolutionally conserved Foxo binding sites within the hepcidin promoter region. These binding sites were found to colocalize with Smad-binding sites. To investigate the physiological relevance of Foxo1 in iron metabolism, we generated mice with hepatocyte-specific deletion of Foxo1. These mice exhibited reduced hepatic hepcidin expression and serum hepcidin levels, accompanied by elevated serum iron and liver non-heme iron concentrations. Moreover, high-iron diet further exacerbated these abnormalities in iron metabolism in mice lacking hepatic Foxo1. Conversely, hepatocyte-specific Foxo1 overexpression increased hepatic hepcidin expression and serum hepcidin levels, thereby ameliorating iron overload in a murine model of hereditary hemochromatosis (Hfe-/- mice). In summary, our study identifies Foxo1 is a critical regulator of hepcidin and systemic iron homeostasis. Targeting Foxo1 may offer therapeutic opportunities for managing conditions associated with aberrant iron metabolism.

Multimodal neuroimaging fusion unravel structural-functional-neurotransmitter change in Parkinson's disease with impulse control disorders.

BACKGROUND: Impulse control disorders (ICD) in Parkinson's disease (PD) is highly multifactorial in etiology and has intricate neural mechanisms. Our multimodal neuroimaging study aimed to investigate the specific patterns of structure-function-neurotransmitter interactions underlying ICD. METHODS: Thirty PD patients with ICD (PD-ICD), 30 without ICD (PD-NICD) and 32 healthy controls (HCs) were recruited. Gyrification and perivascular spaces (PVS) were computed to capture the alternations of cortical surface morphology and glymphatic function. Seed-based functional connectivity (FC) were performed to identify the corresponding functional changes. Further, JuSpace toolbox were employed for cross-modal correlations to evaluate whether the spatial patterns of functional alterations in ICD patients were associated with specific neurotransmitter system. RESULTS: Compared to PD-NICD, PD-ICD patients showed hypogyrification and enlarged PVS volume fraction in the left orbitofrontal gyrus (OFG), as well as decreased FC between interhemispheric OFG. The interhemispheric OFG connectivity reduction was associated with spatial distribution of µ-opioid pathway (r = -0.186, p = 0.029, false discovery rate corrected). ICD severity was positively associated with the PVS volume fraction of left OFG (r = 0.422, p = 0.032). Furthermore, gyrification index (LGI) and percent PVS (pPVS) in OFG and their combined indicator showed good performance in differentiating PD-ICD from PD-NICD. CONCLUSIONS: Our findings indicated that the co-altered structure-function-neurotransmitter interactions of OFG might be involved in the pathogenesis of ICD.

Efficacy and Safety of 5-Aminolevulinic Acid Photodynamic Therapy for Treating Cervical and Vaginal Intraepithelial Neoplasia.

Persistent HPV infections may cause cervical and vaginal intraepithelial neoplasia (CIN and VaIN). Traditional methods might destroy the structure and function of the cervix. 5-aminolevulinic acid photodynamic therapy (ALA-PDT) is a non-invasive targeted therapy. This study aims to evaluate the efficacy and safety of ALA-PDT for CIN and VaIN and the clearance of HPV. A retrospective study of 303 patients who confirmed CIN or VaIN and received ALA-PDT was conducted. All the patients were followed up at six and twelve months after treatment and then annually thereafter. The effect was evaluated through HPV genotyping, a cytology test, and colposcopy-directed biopsy if necessary. After ALA-PDT, the remission rates for CIN 2, CIN 3, VaIN 2, and VaIN 3 were 90.6%, 88.5%, 87.3%, and 77.8%. For CIN 1, the remission rate at the six-month follow-up was 93.1%. The total HPV clearance rates were 72.5% at the six-month follow-up and 85.7% at the 12-month follow-up. The most common adverse event was vaginal discharge. No severe adverse effect was observed. ALA-PDT is an effective and safe treatment for all grades of CIN and VaIN and is helpful in clearing HPV with minimal side effects. This treatment may not influence fertility and delivery.

Undisclosed contribution of microbial assemblages selectively enriched by microplastics to the sulfur cycle in the large deep-water reservoir.

The accumulation of microplastics in reservoirs due to river damming has drawn considerable attention due to their potential impacts on elemental biogeochemical cycling at the watershed scale. However, the effects of plastisphere communities on the sulfur cycle in the large deep-water reservoir remain poorly understood. Here, we collected microplastics and their surrounding environmental samples in the water and sediment ecosystems of Xiaowan Reservoir and found a significant spatiotemporal pattern of microplastics and sulfur distribution in this Reservoir. Based on the microbial analysis, plastic-degrading taxa (e.g., Ralstonia, Rhodococcus) involved in the sulfur cycle were enriched in the plastisphere of water and sediment, respectively. Typical thiosulfate oxidizing bacteria Limnobacter acted as keystone species in the plastisphere microbial network. Sulfate, oxidation reduction potential and organic matter drove the variations of the plastisphere. Environmental filtration significantly affected the plastisphere communities, and the deterministic process dominated the community assembly. Furthermore, predicted functional profiles related to sulfur cycling, compound degradation and membrane transport were significantly enriched in the plastisphere. Overall, our results suggest microplastics as a new microbial niche exert different effects in water and sediment environments, and provide insights into the potential impacts of the plastisphere on the sulfur biogeochemical cycle in the reservoir ecosystem.

Increased miR-3074-5p expression promotes M1 polarization and pyroptosis of macrophages via ERα/NLRP3 pathway and induces adverse pregnancy outcomes in mice.

Decidual macrophages (dMϕs) play critical roles in regulation of immune-microhomeostasis at maternal-fetal interface during pregnancy, but the underlying molecular mechanisms are still unclear. In this study, it was found that litter size and fetal weight were significantly reduced, whereas the rate of embryo resorption was increased in miR-3074-5p knock-in (3074-KI) pregnant mice, compared to that of wild-type (WT) pregnant mice. Plasma levels of pro-inflammatory cytokines in 3074-KI pregnant mice were also significantly elevated compared to WT pregnant mice at GD7.5. The quantity of M1-Mϕs in uterine tissues of 3074-KI pregnant mice was significantly increased compared to WT pregnant mice at GD13.5. Estrogen receptor-α (ERα) was validated to be a target of miR-3074-5p. Either miR-3074-5p overexpression or ERα knockdown promoted transcriptional activity of NF-κB/p65, induced M1-polarization and pyroptosis of THP1-derived Mϕs, accompanied with increased intracellular levels of cleaved Caspase-1, cleaved IL-1ß, NLRP3, cleaved GSDMD and ASC aggregation. Furthermore, ERα could not only bind to NLRP3 or ASC directly, but also inhibit the interaction between NLRP3 and ASC. The endometrial miR-3074-5p expression level at the middle secretory stage of repeated implantation failure (RIF) patients was significantly decreased compared to that of control fertile women. These data indicated that miR-3074-5p could promote M1 polarization and pyroptosis of Mϕs via activation of NLRP3 inflammasome by targeting ERα, and the dysregulation of miR-3074-5p expression in dMϕs might damage the embryo implantation and placentation by interfering with inflammatory microenvironment at the maternal-fetal interface during early pregnancy.

Brucella infection combined with Nocardia infection: A case report and literature review.

Human brucellosis is an infectious disease caused by Brucella and is often misdiagnosed for atypical manifestations including fever of unknown origin, headache, weakness, among else. Nocardiosis is a zoonotic disease caused by the genus Nocardia, which usually spreads through the respiratory tract, skin, and digestive tract. Limited research has documented cases of co-infection involving both Brucella and Nocardia pathogens in patients. A 55-year-old male was admitted to our hospital with intermittent high-grade fever. Following sputum and blood cultures, as well as other laboratory examinations, the patient was diagnosed with concurrent brucellosis and nocardiosis. According to recommendations of previous studies and reports, the patient was successively treated with levofloxacin, doxycycline, piperacillin sodium and sulbactam sodium, trimethoprim-sulfamethoxazole, rifampicin, and tigecycline, after which the patient recovered and was discharged. Brucella and Nocardia are both opportunistic pathogens and simultaneous infection of Brucella and Nocardia is relatively rare. If patients continue to experience persistent fever despite receiving empirical antibiotic therapy, it becomes necessary to conduct examinations to identify potential atypical pathogens, including Brucella and Nocardia. Sputum staining, sputum culture, and blood culture are critical auxiliary examinations during clinical practice. The treatment plan should be selected based on guidelines and the individual patient's condition. Regular reevaluation should be conducted, and antimicrobial agents should be adjusted accordingly.

Antioxidant Minerals Modified the Association between Iron and Type 2 Diabetes in a Chinese Population.

Inconsistent findings exist regarding the relationship between heme iron intake and type 2 diabetes (T2D) among Western and Eastern populations. Easterners tend to consume a plant-based diet which is abundant in antioxidant minerals. To examine the hypothesis that antioxidant mineral may modify the relationship between iron and T2D, we performed a case-control study by measuring the serum mineral levels in 2198 Chinese subjects. A total of 2113 T2D patients and 2458 controls were invited; 502 T2D patients and 1696 controls were finally analyzed. In the total population, high serum iron showed a positive association with T2D odds (odds ratio [OR] = 1.27 [1.04, 1.55]); high magnesium (OR = 0.18 [0.14, 0.22]), copper (OR = 0.27 [0.21, 0.33]), zinc (OR = 0.37 [0.30, 0.46]), chromium (OR = 0.61 [0.50, 0.74]), or selenium concentrations (OR = 0.39 [0.31, 0.48]) were inversely associated with T2D odds. In contrast, in individuals with higher magnesium (>2673.2 µg/dL), zinc (>136.7 µg/dL), copper (>132.1 µg/dL), chromium (>14.0 µg/dL), or selenium concentrations (>16.8 µg/dL), serum iron displayed no association with T2D (p > 0.05). Serum copper and magnesium were significant modifiers of the association between iron and T2D in individuals with different physiological status (p < 0.05). Our findings support the idea that consuming a diet rich in antioxidant minerals is an effective approach for preventing T2D.

Association of elastic power in mechanical ventilation with the severity of acute respiratory distress syndrome: a retrospective study.

BACKGROUND: Mechanical power (MP) is the total energy released into the entire respiratory system per minute which mainly comprises three components: elastic static power, Elastic dynamic power and resistive power. However, the energy to overcome resistance to the gas flow is not the key factor in causing lung injury, but the elastic power (EP) which generates the baseline stretch of the lung fibers and overcomes respiratory system elastance may be closely related to the ARDS severity. Thus, this study aimed to investigate whether EP is superior to other ventilator variables for predicting the severity of lung injury in ARDS patients. METHODS: We retrieved patient data from the Medical Information Mart for Intensive Care III (MIMIC-III) database. The retrieved data involved adults (≥ 18 years) diagnosed with ARDS and subjected to invasive mechanical ventilation for ≥ 48 h. We employed univariate and multivariate logistic regression analyses to investigate the correlation between EP and development of moderate-severe ARDS. Furthermore, we utilized restricted cubic spline models to assess whether there is a linear association between EP and incidence of moderate-severe ARDS. In addition, we employed a stratified linear regression model and likelihood ratio test in subgroups to identify potential modifications and interactions. RESULTS: Moderate-severe ARDS occurred in 73.4% (296/403) of the patients analyzed. EP and MP were significantly associated with moderate-severe ARDS (odds ratio [OR] 1.21, 95% confidence interval [CI] 1.15-1.28, p < 0.001; and OR 1.15, 95%CI 1.11-1.20, p < 0.001; respectively), but EP showed a higher area-under-curve (95%CI 0.72-0.82, p < 0.001) than plateau pressure, driving pressure, and static lung compliance in predicting ARDS severity. The optimal cutoff value for EP was 14.6 J/min with a sensitivity of 75% and specificity of 66%. Quartile analysis revealed that the relationship between EP and ARDS severity remained robust and reliable in subgroup analysis. CONCLUSION: EP is a good ventilator variable associated with ARDS severity and can be used for grading ARDS severity. Close monitoring of EP is advised in patients undergoing mechanical ventilation. Additional experimental trials are needed to investigate whether adjusting ventilator variables according to EP can yield significant improvements in clinical outcomes.

The Cd immobilization mechanisms in paddy soil through ureolysis-based microbial induced carbonate precipitation: Emphasis on the coexisting cations and metatranscriptome analysis.

Microbial induced carbonate precipitation (MICP) can immobilize metals and reduce their bioavailability. However, little is known about the immobilization mechanism of Cd in the presence of soil cations and the triggered gene expression and metabolic pathways in paddy soil. Thus, microcosmic experiments were conducted to study the fractionation transformation of Cd and metatranscriptome analysis. Results showed that bioavailable Cd decreased from 0.62 to 0.29 mg/kg after 330 d due to the MICP immobilization. This was ascribed to the increase in carbonate bound, Fe-Mn oxides bound, and residual Cd. The underlying immobilization mechanisms could be attributed to the formation of insoluble Cd-containing precipitates, the complexation and lattice substitution with carbonate and Fe, Mn and Al (hydr)oxides, and the adsorption on functional group on extracellular polymers of cell. During the MICP immobilization process, up-regulated differential expression urease genes were significantly enriched in the paddy soil, corresponding to the arginine biosynthesis, purine metabolism and atrazine degradation. The metabolic pathway of bacterial chemotaxis, flagellum assembly, and peptidoglycan biosynthesis and the expression of cadA gene related to Cd excretion enhanced Cd resistance of soil microbiome. Therefore, this study provided new insights into the immobilization mechanisms of Cd in paddy soils through ureolysis-based MICP process.

Human Motion Generation: A Survey.

Human motion generation aims to generate natural human pose sequences and shows immense potential for real-world applications. Substantial progress has been made recently in motion data collection technologies and generation methods, laying the foundation for increasing interest in human motion generation. Most research within this field focuses on generating human motions based on conditional signals, such as text, audio, and scene contexts. While significant advancements have been made in recent years, the task continues to pose challenges due to the intricate nature of human motion and its implicit relationship with conditional signals. In this survey, we present a comprehensive literature review of human motion generation, which, to the best of our knowledge, is the first of its kind in this field. We begin by introducing the background of human motion and generative models, followed by an examination of representative methods for three mainstream sub-tasks: text-conditioned, audio-conditioned, and scene-conditioned human motion generation. Additionally, we provide an overview of common datasets and evaluation metrics. Lastly, we discuss open problems and outline potential future research directions. We hope that this survey could provide the community with a comprehensive glimpse of this rapidly evolving field and inspire novel ideas that address the outstanding challenges.

Transport Behavior of Cd2+ in Highly Weathered Acidic Soils and Shaping in Soil Microbial Community Structure.

The mining and smelting site soils in South China present excessive Cd pollution. However, the transport behavior of Cd in the highly weathered acidic soil layer at the lead-zinc smelting site remains unclear. Here, under different conditions of simulated infiltration, the migration behavior of Cd2+ in acid smelting site soils at different depths was examined. The remodeling effect of Cd2+ migration behavior on microbial community structure and the dominant microorganisms in lead-zinc sites soils was analyzed using high-throughput sequencing of 16S rRNA gene amplicons. The results revealed a specific flow rate in the range of 0.3-0.5 mL/min that the convection and dispersion have no obvious effect on Cd2+ migration. The variation of packing porosity could only influence the migration behavior by changing the average pore velocity, but cannot change the adsorption efficiency of soil particles. The Cd has stronger migration capacity under the reactivation of acidic seepage fluid. However, in the alkaline solution, the physical properties of soil, especially pores, intercept the Cd compounds, further affecting their migration capacity. The acid-site soil with high content of SOM, amorphous Fe oxides, crystalline Fe/Mn/Al oxides, goethite, and hematite has stronger ability to adsorb and retain Cd2+. However, higher content of kaolinite in acidic soil will increase the potential migration of Cd2+. Besides, the migration behavior of Cd2+ results in simplified soil microbial communities. Under Cd stress, Cd-tolerant genera (Bacteroides, Sphingomonas, Bradyrhizobium, and Corynebacterium) and bacteria with both acid-Cd tolerance (WCHB 1-84) were distinguished. The Ralstonia showed a high enrichment degree in alkaline Cd2+ infiltration solution (pH 10.0). Compared to the influence of Cd2+ stress, soil pH had a stronger ability to shape the microbial community in the soil during the process of Cd2+ migration.

MiR-3074-5p Regulates Trophoblasts Function via EIF2S1/GDF15 Pathway in Recurrent Miscarriage.

Dysfunction of extravillous trophoblasts (EVTs) might cause early pregnancy failure by interfering with embryo implantation and/or placentation. We previously reported that the villus miR-3074-5p expression level was increased, whereas the peripheral level of GDF15, a predict target gene of miR-3074-5p, was decreased in recurrent miscarriages (RM) patients, and miR-3074-5p could enhance apoptosis but reduce invasion of human extravillous trophoblast cells (EVTs). The aim of this study was to further explore roles of miR-3074-5p/GDF15 pathway in regulation of EVTs function. It was validated that GDF15 was not the direct target of miR-3074-5p, whereas EIF2S1, an upstream regulator of GDF15 maturation and secretion, was the direct target of miR-3074-5p. The villus expression levels of GDF15 and EIF2S1 were significantly decreased in RM patients. Knockdown of GDF15 expression presented inhibitory effects on proliferation, migration, and invasion of HTR8/SVneo cells. Up-regulated miR-3074-5p expression led to the significant decreased GDF15 expression in HTR8/SVneo cells, and this effect could be efficiently reversed by the overexpression of EIF2S1. Meanwhile, the suppressive effects of miR-3074-5p on proliferation, migration, and invasion of HTR8/SVneo cells could be intercepted by the treatment of recombinant human GDF15 protein. Collectively, these data suggested that miR-3074-5p could reduce GDF15 production via targeting inhibition of EIF2S1 expression, and the deficiency in GDF15 function might lead to the early pregnancy loss by attenuating proliferation and invasion of EVTs.

Using Radiomics to Differentiate Brain Metastases From Lung Cancer Versus Breast Cancer, Including Predicting Epidermal Growth Factor Receptor and human Epidermal Growth Factor Receptor 2 Status.

OBJECTIVE: We evaluated the feasibility of using multiregional radiomics to identify brain metastasis (BM) originating from lung adenocarcinoma (LA) and breast cancer (BC) and assess the epidermal growth factor receptor (EGFR) mutation and human epidermal growth factor receptor 2 (HER2) status. METHODS: Our experiment included 160 patients with BM originating from LA (n = 70), BC (n = 67), and other tumor types (n = 23), between November 2017 and December 2021. All patients underwent contrast-enhanced T1- and T2-weighted magnetic resonance imaging (MRI) scans. A total of 1967 quantitative MRI features were calculated from the tumoral active area and peritumoral edema area and selected using least absolute shrinkage and selection operator regression with 5-fold cross-validation. We constructed radiomic signatures (RSs) based on the most predictive features for preoperative assessment of the metastatic origins, EGFR mutation, and HER2 status. Prediction performance of the constructed RSs was evaluated based on the receiver operating characteristic curve analysis. RESULTS: The developed multiregion RSs generated good area under the receiver operating characteristic curve (AUC) for identifying the LA and BC origin in the training (AUCs, RS-LA vs RS-BC, 0.767 vs 0.898) and validation (AUCs, RS-LA vs RS-BC, 0.778 and 0.843) cohort and for predicting the EGFR and HER2 status in the training (AUCs, RS-EGFR vs RS-HER2, 0.837 vs 0.894) and validation (AUCs, RS-EGFR vs RS-HER2, 0.729 vs 0.784) cohorts. CONCLUSIONS: Our results revealed associations between brain MRI-based radiomics and their metastatic origins, EGFR mutations, and HER2 status. The developed multiregion combined RSs may be considered noninvasive predictive markers for planning early treatment for BM patients.

Leaching Behavior of As and Pb in Lead-Zinc Mining Waste Rock under Mine Drainage and Rainwater.

At present, the pollution of arsenic (As) and lead (Pb) is becoming increasingly serious. The pollution caused by the release of As and Pb from lead-zinc mines has seriously affected the water and soil environment and threatened human health. It is necessary to reveal the release characteristics of As and Pb. The actual scene of mine drainage (MD) and rainwater (RW) leaching waste rocks is the one of the main reasons for the release of As and Pb. However, the leaching behavior of As and Pb in these waste rocks under MD and RW suffered from a lack of in-depth research. In this study, we investigated the occurrence of As and Pb in waste rocks (S1-S6) by using X-ray diffraction (XRD) and time-of-flight secondary ion mass spectrometry (TOF-SIMS), and then, the changes in As and Pb concentration and the hydrochemical parameter in leaching solution were systematically studied. Furthermore, the correlation between the release of As and Pb and mineral composition was also evaluated. Results showed that these waste rocks were mainly composed of carbonate and sulfide minerals. As and Pb were mainly bounded or associated with sulfide minerals such as arsenopyrite, pyrite, chalcopyrite, and galena in these waste rocks, and small parts of As and Pb were absorbed or encased by clay minerals such as kaolinite and chlorite. Under MD and RW leaching, the pH, redox potential (Eh), and electric conductivity (EC) of each waste rock tended to be consistent due to their buffering ability; the leachate pH of waste rocks with more carbonate minerals was higher than that of sulfide minerals. Both As and Pb were released most under MD leaching in comparison to RW, reaching 6.57 and 60.32 mg/kg, respectively, due to MD's low pH and high Eh value. However, As in waste rock released more under alkaline conditions because part of the arsenic was in the form of arsenate. As and Pb release were mainly positively correlated with the proportions of sulfide minerals in these waste rocks. MD leaching significantly promoted the release of As and Pb from waste rocks, which would cause a great threat to the surrounding environment, and control measures were imperative. This paper not only reveals the As and Pb pollution mechanism around the lead-zinc mining area but also provides a theoretical basis for the prevention and control of As and Pb pollution in the future.

Elucidating the impact of in vitro cultivation on Nicotiana tabacum metabolism through combined in silico modeling and multiomics analysis.

The systematical characterization and understanding of the metabolic behaviors are the basis of the efficient plant metabolic engineering and synthetic biology. Genome-scale metabolic networks (GSMNs) are indispensable tools for the comprehensive characterization of overall metabolic profile. Here we first constructed a GSMN of tobacco, which is one of the most widely used plant chassis, and then combined the tobacco GSMN and multiomics analysis to systematically elucidate the impact of in-vitro cultivation on the tobacco metabolic network. In-vitro cultivation is a widely used technique for plant cultivation, not only in the field of basic research but also for the rapid propagation of valuable horticultural and pharmaceutical plants. However, the systemic effects of in-vitro cultivation on overall plant metabolism could easily be overlooked and are still poorly understood. We found that in-vitro tobacco showed slower growth, less biomass and suppressed photosynthesis than soil-grown tobacco. Many changes of metabolites and metabolic pathways between in-vitro and soil-grown tobacco plants were identified, which notably revealed a significant increase of the amino acids content under in-vitro condition. The in silico investigation showed that in-vitro tobacco downregulated photosynthesis and primary carbon metabolism, while significantly upregulated the GS/GOGAT cycle, as well as producing more energy and less NADH/NADPH to acclimate in-vitro growth demands. Altogether, the combination of experimental and in silico analyses offers an unprecedented view of tobacco metabolism, with valuable insights into the impact of in-vitro cultivation, enabling more efficient utilization of in-vitro techniques for plant propagation and metabolic engineering.

The impacts of casual environmental regulation on carbon intensity in China: Dual mediating pathways of energy low-carbon reconstitution and industrial structure upgrading.

In the new era of global net zero, how to reduce carbon intensity became a hot topic, while few attentions had been paid to the impacts of casual environmental regulation, especially in the emerging economy such as China. In this regard, this study uses the panel data of 30 provinces in China from 2003 to 2019 to study the impacts of causal environmental regulation on the carbon intensity, with the adoption of multiple econometric models. The empirical results show that causal environmental regulation has a significant inhibitory effect on carbon intensity, while at current stage, causal environmental regulation cannot inhibit carbon intensity through the mediating of green innovation and renewable energy production. Fortunately, energy low-carbon reconstitution and industrial structure upgrading are proved to effectively act as the intermediary channels. In addition, the spatial spillover effects and the spatial heterogeneity are supported in the empirical analysis. In a summary, this study starts with the case of China to provide a realistic reference case for the international community, enriches the theory, policy and practice related to environmental regulation, and provides a realistic reference example for emerging economics in a developing country.

Green tissue-targeted expression of the Cry1Ab/c gene in transgenic maize using the Cre/loxP system as an alternative strategy against lepidopteran pests.

Genetically modified (GM) proteins in edible tissues of transgenic maize are of intense public concern. We provided a Cre/loxP-based strategy for manipulating the expression of transgenes in green tissues while locking them in nongreen tissues. First, the Cre gene was driven by the green tissue-specific promoter Zm1rbcS to generate transgenic maize KEY. Meanwhile, a gene cassette containing a Nos terminator (NosT) in front of the Cry1Ab/c gene was driven by the strong promoter ZmUbi to generate another transgenic maize LOCK. By crossing KEY and LOCK plants, the expressed Cre recombinase under the control of the Zm1rbcS promoter from KEY maize accurately removed the NosT of LOCK maize. Consequently, the expression of blocked Cry1Ab/c was enabled in specific green tissues in their hybrids. The expression level and concentration of Cry1Ab/c were observed using a strategy with high specific accumulation in green tissues (leaf and stem). Still, only a small or absent amount was observed in root and kernel tissues. Furthermore, we assessed the bioactivity of transgenic maize against 2 common lepidopteran pests, Ostrinia furnacalis and Spodoptera frugiperda, in the laboratory and field. The transgenic plants showed high plant resistance levels against the 2 pests, with mortality rates above 97.2% and damage scales below 2.2 compared with the control group. These findings are significant for exploring novel genetic engineering techniques in GM maize and providing a feasible strategy for transgenes avoiding expression in edible parts. In addition, implementing the Cre/loxP-mediated system could relieve public sentiment toward the biosafety of GM plants.