Transcriptomics and metabolomics analysis of L-phenylalanine overproduction in Escherichia coli.

BACKGROUND: Highly efficient production of L-phenylalanine (L-Phe) in E. coli has been achieved by multiple rounds of random mutagenesis and modification of key genes of the shikimate (SHIK) and L-Phe branch pathways. In this study, we performed transcriptomic (16, 24 and 48 h) and metabolomic analyses (8, 16, 24, 32,40, and 48 h) based on time sequences in an engineered E. coli strain producing L-Phe, aiming to reveal the overall changes of metabolic activities during the fermentation process. RESULTS: The largest biomass increase rate and the highest production rate were seen at 16 h and 24 h of fermentation, respectively reaching 5.9 h-1 and 2.76 g/L/h, while the maximal L-Phe titer of 60 g/L was accumulated after 48 h of fermentation. The DEGs and metabolites involved in the EMP, PP, TCA, SHIIK and L-Phe-branch pathways showed significant differences at different stages of fermentation. Specifically, the significant upregulation of genes encoding rate-limiting enzymes (aroD and yidB) and key genes (aroF, pheA and aspC) pushed more carbon flux toward the L-Phe synthesis. The RIA changes of a number of important metabolites (DAHP, DHS, DHQ, Glu and PPN) enabled the adequate supply of precursors for high-yield L-Phe production. In addition, other genes related to Glc transport and phosphate metabolism increased the absorption of Glc and contributed to rerouting the carbon flux into the L-Phe-branch. CONCLUSIONS: Transcriptomic and metabolomic analyses of an L-Phe overproducing strain of E. coli confirmed that precursor supply was not a major limiting factor in this strain, whereas the rational distribution of metabolic fluxes was achieved by redistributing the carbon flux (for example, the expression intensity of the genes tyrB, aspC, aroL and aroF/G/H or the activity of these enzymes is increased to some extent), which is the optimal strategy for enhancing L-Phe production.

Ultra-short echo time MR imaging in assessing cartilage endplate damage and relationship between its lesion and disc degeneration for chronic low back pain patients.

OBJECTIVE: To investigate the feasibility of ultra-short echo time (UTE) magnetic resonance imaging (MRI) in the assessment of cartilage endplate (CEP) damage and further evaluate the relationship between total endplate score (TEPS) and lumbar intervertebral disc (IVD) degeneration for chronic low back pain patients. MATERIALS AND METHODS: IVD were measured in 35 patients using UTE imaging at 3T MR. Subtracted UTE images between short and long TEs were obtained to depict anatomy of CEP. The signal-to-noise ratio (SNR) and contrast-to-noise ratio (CNR) were calculated to assess the image quality quantitatively. A new grading criterion for endplate damage evaluation was developed based on Rajasekaran.S grading system in this study. Two radiologists were employed to evaluate CEP and bony vertebral endplates (VEP) using this new grading criterion and assess TEPS, independently. Cohen's kappa analysis was applied to evaluate the inter-observer agreement of endplate damage assessment between two radiologists, and the Kendall's TAU-B analysis was employed to determine the relationship between TEPS and IVD degeneration evaluated with Pfirrmann grading. RESULTS: Well structural CEP was depicted on subtracted UTE images and confirmed by high SNR (33.06±2.92) and CNR values (9.4±2.08). Qualified subtracted UTE images were used by two radiologists to evaluate the degree of CEP and VEP damage. Excellent inter-observer agreement was confirmed by high value in Cohen's kappa test (0.839, P < 0.001). Ensured by this, 138 endplates from 69 IVDs of 35 patients were classified into six grades based on the new grading criterion and TEPS of each endplate was calculated. In addition, the degeneration degree of IVDs were classified into five grades. Finally, using Kendall's TAU-B analysis, significant relationship was obtained between endplate damage related TEPS and IVD degeneration (r = 0.864, P < 0.001). CONCLUSION: Ensured by high image quality, UTE imaging might be considered an effective tool to assess CEP damage. Additionally, further calculated TEPS has shown strong positive association with IVD degeneration, suggesting that the severity of endplate damage is highly linked with the degree of IVD degeneration.

Relationships among patient activation, social support and online health information seeking of community-dwelling older adults living with coronary heart disease.

AIMS: To explore the relationship between patient activation (subjective initiative of patients to participate in disease management in self-health care), social support, self-efficacy and online health information seeking among older patients with coronary heart disease. DESIGN: A cross-sectional and survey-based design. METHODS: A total of 451 older patients with coronary heart disease were recruited from July to November 2021 from four communities in Qingdao, China. We collected data using the Patient Activation Measure, Social Support Rating Scale, Self-Efficacy Scale and Online Health Information Seeking Scale. We performed structural equation modelling to analyse the data. RESULTS: The final model showed good model fit. Patient activation influenced online health information seeking directly (ß  = .39, p < .05) and indirectly through self-efficacy (ß  = .17, p < .05). Social support influenced online health information seeking directly (ß  = .23, p < .05) and indirectly through self-efficacy (ß  = .03, p < .05). Self-efficacy directly influenced online health information seeking (ß  = .26, p < .05). CONCLUSIONS: We identified the interrelationships of patient activation, social support and self-efficacy and their influence on the online health information seeking of older patients with coronary heart disease. Our findings provide a theoretical basis for developing and evaluating interventions to enhance online health information seeking for older patients with coronary heart disease. IMPACT: These findings add a better understanding of the relationship between patient activation, social support, self-efficacy and online health information seeking in older patients with coronary heart disease, and help community health workers to intervene in the early stage of disease diagnosis.

Bioprinted PDLSCs with high-concentration GelMA hydrogels exhibit enhanced osteogenic differentiation in vitro and promote bone regeneration in vivo.

OBJECTIVES: We aimed to explore the osteogenic potential of periodontal ligament stem cells (PDLSCs) in bioprinted methacrylate gelatine (GelMA) hydrogels in vitro and in vivo. MATERIALS AND METHODS: PDLSCs in GelMA hydrogels at various concentrations (3%, 5%, and 10%) were bioprinted. The mechanical properties (stiffness, nanostructure, swelling, and degradation properties) of bioprinted constructs and the biological properties (cell viability, proliferation, spreading, osteogenic differentiation, and cell survival in vivo) of PDLSCs in bioprinted constructs were evaluated. Then, the effect of bioprinted constructs on bone regeneration was investigated using a mouse cranial defect model. RESULTS: Ten percent GelMA printed constructs had a higher compression modulus, smaller porosity, lower swelling rate, and lower degradation rate than 3% GelMA. PDLSCs in bioprinted 10% GelMA bioprinted constructs showed lower cell viability, less cell spreading, upregulated osteogenic differentiation in vitro, and lower cell survival in vivo. Moreover, upregulated expression of ephrinB2 and EphB4 protein and their phosphorylated forms were found in PDLSCs in 10% GelMA bioprinted constructs, and inhibition of eprhinB2/EphB4 signalling reversed the enhanced osteogenic differentiation of PDLSCs in 10% GelMA. The in vivo experiment showed that 10% GelMA bioprinted constructs with PDLSCs contributed to more new bone formation than 10% GelMA constructs without PDLSCs and constructs with lower GelMA concentrations. CONCLUSIONS: Bioprinted PDLSCs with high-concentrated GelMA hydrogels exhibited enhanced osteogenic differentiation partially through upregulated ephrinB2/EphB4 signalling in vitro and promoted bone regeneration in vivo, which might be more appropriate for future bone regeneration applications. CLINICAL RELEVANCE: Bone defects are a common clinical oral problem. Our results provide a promising strategy for bone regeneration through bioprinting PDLSCs in GelMA hydrogels.

Virus Infection and mRNA Nuclear Export.

Gene expression in eukaryotes begins with transcription in the nucleus, followed by the synthesis of messenger RNA (mRNA), which is then exported to the cytoplasm for its translation into proteins. Along with transcription and translation, mRNA export through the nuclear pore complex (NPC) is an essential regulatory step in eukaryotic gene expression. Multiple factors regulate mRNA export and hence gene expression. Interestingly, proteins from certain types of viruses interact with these factors in infected cells, and such an interaction interferes with the mRNA export of the host cell in favor of viral RNA export. Thus, these viruses hijack the host mRNA nuclear export mechanism, leading to a reduction in host gene expression and the downregulation of immune/antiviral responses. On the other hand, the viral mRNAs successfully evade the host surveillance system and are efficiently exported from the nucleus to the cytoplasm for translation, which enables the continuation of the virus life cycle. Here, we present this review to summarize the mechanisms by which viruses suppress host mRNA nuclear export during infection, as well as the key strategies that viruses use to facilitate their mRNA nuclear export. These studies have revealed new potential antivirals that may be used to inhibit viral mRNA transport and enhance host mRNA nuclear export, thereby promoting host gene expression and immune responses.

Na2[(VO)2(HPO4)2(C2O4)]·2H2O: A Promising Mixed Polyanionic Cathode Material for Aqueous Zn-Ion Batteries.

A cathode material, Na2[(VO)2(HPO4)2(C2O4)]·2H2O, for aqueous zinc-ion batteries is synthesized by a hydrothermal method. Na2[(VO)2(HPO4)2(C2O4)]·2H2O presents a two-dimensional layered structure and sheetlike morphology, which provide fast and reversible Zn2+ insertion/extraction. Na2[(VO)2(HPO4)2(C2O4)]·2H2O delivers a high average voltage plateau (∼1.3 V), a moderate specific capacity (90 mA h g-1), and good cycle stability (100% after 100 cycles). The Zn2+ storage mechanism is investigated by in situ XRD and ex situ XPS.

Relationships among social support, self-efficacy, and patient activation in community-dwelling older adults living with coronary heart disease: A cross-sectional study.

OBJECTIVES: To explore whether social support indirectly influences patient activation through self-efficacy in older adults living with coronary heart disease. METHODS: A cross-sectional study was conducted. Older patients (n=451) from four communities in the city of Qingdao completed a questionnaire survey. We conducted multiple linear regression models and bootstrap testing to assess the relationships among social support, self-efficacy, and patient activation. RESULTS: Patient activation was positively correlated with social support (r = 0.524, P < 0.01) and with self-efficacy (r = 0.740, P < 0.01). The of social support had indirect positive effect on patient activation through self-efficacy and the effect was 58.8%. CONCLUSIONS: We identified the critical role of social support and self-efficacy for the activation of community-dwelling older patients living with coronary heart disease. Our findings provide essential knowledge for developing and evaluating effective interventions to promote patient activation and enhance self-management of coronary heart disease.

The polycomb group protein PCGF6 mediates germline gene silencing by recruiting histone-modifying proteins to target gene promoters.

Polycomb group (PcG) proteins are essential for maintenance of lineage fidelity by coordinating developmental gene expression programs. Polycomb group ring finger 6 (PCGF6) has been previously reported to repress expression of lineage-specific genes, especially germ cell-related genes in mouse embryonic stem cells (ESCs) via the noncanonical polycomb repressive complex PRC1.6. However, the molecular mechanism of this repression remains largely unknown. Here, using RNA-Seq, real-time RT-PCR, immunohistochemistry, immunoprecipitation, and ChIP analyses, we demonstrate that PCGF6 plays an essential role in embryonic development, indicated by the partially penetrant embryonic lethality in homozygous PCGF6 (Pcgf6-/-)-deficient mice. We also found that surviving Pcgf6-deficient mice exhibit reduced fertility. Using the Pcgf6-deficient mice, we observed that ablation of Pcgf6 in somatic tissues robustly derepresses germ cell-related genes. We further provide evidence that these genes are direct targets of PCGF6 in ESCs and that endogenous PCGF6 co-localizes with the histone-modifying proteins G9A histone methyltransferase (G9A)/G9a-like protein (GLP) and histone deacetylase 1/2 (HDAC1/2) on the promoters of the germ cell-related genes. Moreover, the binding of these proteins to their target genes correlated with methylation of Lys-9 of histone 3 and with the status of histone acetylation at these genes. Moreover, the recruitment of G9A/GLP and HDAC1/2 to target promoters depended on the binding of PCGF6. Our findings indicate that PCGF6 has a critical role in safeguarding lineage decisions and in preventing aberrant expression of germ cell-related genes.

Analyzing the genetic characteristics of a tryptophan-overproducing Escherichia coli.

L-tryptophan (L-trp) production in Escherichia coli has been developed by employing random mutagenesis and selection for a long time, but this approach produces an unclear genetic background. Here, we generated the L-trp overproducer TPD5 by combining an intracellular L-trp biosensor and fluorescence-activated cell sorting (FACS) in E. coli, and succeeded in elucidating the genetic basis for L-trp overproduction. The most significant identified positive mutations affected TnaA (deletion), AroG (S211F), TrpE (A63V), and RpoS (nonsense mutation Q33*). The underlying structure-function relationships of the feedback-resistant AroG (S211F) and TrpE (A63V) mutants were uncovered based on protein structure modeling and molecular dynamics simulations, respectively. According to transcriptomic analysis, the global regulator RpoS not only has a great influence on cell growth and morphology, but also on carbon utilization and the direction of carbon flow. Finally, by balancing the concentrations of the L-trp precursors' serine and glutamine based on the above analysis, we further increased the titer of L-trp to 3.18 g/L with a yield of 0.18 g/g. The analysis of the genetic characteristics of an L-trp overproducing E. coli provides valuable information on L-trp synthesis and elucidates the phenotype and complex cellular properties in a high-yielding strain, which opens the possibility to transfer beneficial mutations and reconstruct an overproducer with a clean genetic background.

FGFRL1 affects chemoresistance of small-cell lung cancer by modulating the PI3K/Akt pathway via ENO1.

Fibroblast growth factor receptor-like 1 (FGFRL1), a member of the FGFR family, has been demonstrated to play important roles in various cancers. However, the role of FGFRL1 in small-cell lung cancer (SCLC) remains unclear. Our study aimed to investigate the role of FGFRL1 in chemoresistance of SCLC and elucidate the possible molecular mechanism. We found that FGFRL1 levels are significantly up-regulated in multidrug-resistant SCLC cells (H69AR and H446DDP) compared with the sensitive parental cells (H69 and H446). In addition, clinical samples showed that FGFRL1 was overexpressed in SCLC tissues, and high FGFRL1 expression was associated with the clinical stage, chemotherapy response and survival time of SCLC patients. Knockdown of FGFRL1 in chemoresistant SCLC cells increased chemosensitivity by increasing cell apoptosis and cell cycle arrest, whereas overexpression of FGFRL1 in chemosensitive SCLC cells produced the opposite results. Mechanistic investigations showed that FGFRL1 interacts with ENO1, and FGFRL1 was found to regulate the expression of ENO1 and its downstream signalling pathway (the PI3K/Akt pathway) in SCLC cells. In brief, our study demonstrated that FGFRL1 modulates chemoresistance of SCLC by regulating the ENO1-PI3K/Akt pathway. FGFRL1 may be a predictor and a potential therapeutic target for chemoresistance in SCLC.

The polycomb group protein Yaf2 regulates the pluripotency of embryonic stem cells in a phosphorylation-dependent manner.

The polycomb group (PcG) proteins are key epigenetic regulators in stem cell maintenance. PcG proteins have been thought to act through one of two polycomb repressive complexes (PRCs), but more recent biochemical analyses have challenged this model in the identification of noncanonical PRC1 (nc-PRC1) complexes characterized by the presence of Rybp or Yaf2 in place of the canonical Chromobox proteins. However, the biological significance of these nc-PRC1s and the potential mechanisms by which they mediate gene repression are largely unknown. Here, we explore the functional consequences of Yaf2 disruption on stem cell regulation. We show that deletion of Yaf2 results in compromised proliferation and abnormal differentiation of mouse embryonic stem cells (mESCs). Genome-wide profiling indicates Yaf2 functions primarily as a transcriptional repressor, particularly impacting genes associated with ectoderm cell fate in a manner distinct from Rybp. We confirm that Yaf2 assembles into a noncanonical PRC complex, with deletion analysis identifying the region encompassing amino acid residues 102-150 as required for this assembly. Furthermore, we identified serine 166 as a Yaf2 phosphorylation site, and we demonstrate that mutation of this site to alanine (S166A) compromises Ring1B-mediated H2A monoubiquitination and in turn its ability to repress target gene expression. We therefore propose that Yaf2 and its phosphorylation status serve as dual regulators to maintain the pluripotent state in mESCs.

Stereotactic Body Radiotherapy Versus Surgery for Early-Stage Non-Small-Cell Lung Cancer.

BACKGROUND: Surgery is the gold standard therapy for patients with early-stage non-small-cell lung cancer (NSCLC). However, stereotactic body radiotherapy (SBRT) may provide as an alternative for patients who are medically inoperable or refuse surgical resection. The optimal treatment (SBRT or surgery) for patients with early-stage NSCLC is not clear. METHODS: A systematic search was performed from PubMed, MEDLINE, Embase, and the Cochrane Library. Study heterogeneity and publication bias were estimated. RESULTS: Fourteen cohort studies involving 1438 participants (719 who received SBRT and 719 who received surgery) were included in the meta-analysis. The main bias sources between the two groups, such as age, gender, tumor diameter, forced expiratory volume in 1 s, and Charlson comorbidity index were matched. The surgery was associated with a better overall survival (OS) and long-term distant control (DC) for early-stage NSCLC. The pooled OR and 95% confidence interval (CI) for 1-y, 3-y, 5-y OS, and 5-y DC were 1.56 (1.12-2.15), 1.86 (1.50-2.31), 2.43 (1.80-3.28), and 2.74 (1.12-6.67), respectively. No difference was found between the treatments in the 1-y and 3-y disease-free survival; 1-y, 3-y and 5-y locoregional control; or 1-y and 3-y DC. CONCLUSIONS: Our results found a superior OS and long-term DC for early-stage NSCLC after surgery compared with SBRT after propensity score matching.

Feeding the enemy: loss of nectar and nectaries to herbivores reduces tepal damage and increases pollinator attraction in Iris bulleyana.

Floral nectar usually functions as a pollinator reward, yet it may also attract herbivores. However, the effects of herbivore consumption of nectar or nectaries on pollination have rarely been tested. We investigated Iris bulleyana, an alpine plant that has showy tepals and abundant nectar, in the Hengduan Mountains of SW China. In this region, flowers are visited mainly by pollen-collecting pollinators and nectarivorous herbivores. We tested the hypothesis that, in I. bulleyana, sacrificing nectar and nectaries to herbivores protects tepals and thus enhances pollinator attraction. We compared rates of pollination and herbivory on different floral tissues in plants with flowers protected from nectar and nectary consumption with rates in unprotected control plants. We found that nectar and nectaries suffered more herbivore damage than did tepals in natural conditions. However, the amount of tepal damage was significantly greater in the flowers with protected nectaries than in the controls; this resulted in significant differences in pollinator visitation rates. These results provide the first evidence that floral nectar and nectaries may be 'sacrificed' to herbivores, leading to reduced damage to other floral tissues that are more important for reproduction.

3D bioprinting of DPSCs with GelMA hydrogel of various concentrations for bone regeneration.

Bioprinting technology promotes innovation of fabricating tissue engineered constructs. Dental pulp stem cells (DPSCs) have significant advantages over classical bone mesenchymal stem cells (BMSCs) and are a promising seed cell candidate for bone engineering bioprinting. However, current reports about bioprinted DPSCs for bone regeneration are incomprehensive. The objective of this study was to investigate the osteogenic potential of DPSCs in methacrylate gelatin (GelMA) hydrogels bioprinted scaffolds in vitro and in vivo. Firstly, we successfully bioprinted GelMA with different concentrations embedded with or without DPSCs. Printability, physical features and biological properties of the bioprinted constructs were evaluated. Then, osteogenic differentiation levels of DPSCs in bioprinted constructs with various concentrated GelMA were compared. Finally, effects of bioprinted constructs on cranial bone regeneration were evaluated in vivo. The results of our study demonstrated that 10% GelMA had higher compression modulus, smaller pores, lower swelling and degradation rate than 3% GelMA. Twenty-eight days after printing, DPSCs in three groups of bioprinted structures still maintained high cell activities (>90%). Moreover, DPSCs in 10% GelMA showed an upregulated expression of osteogenic markers and a highly activated ephrinB2/EphB4 signaling, a signaling involved in bone homeostasis. In vivo experiments showed that DPSCs survived at a higher rate in 10% GelMA, and more new bones were observed in DPSC-laden 10% GelMA group, compared with GelMA of other concentrations. In conclusion, bioprinted DPSC-laden 10% GelMA might be more appropriate for bone regeneration application, in contrast to GelMA with other concentrations.

Strategies of functionalized GelMA-based bioinks for bone regeneration: Recent advances and future perspectives.

Gelatin methacryloyl (GelMA) hydrogels is a widely used bioink because of its good biological properties and tunable physicochemical properties, which has been widely used in a variety of tissue engineering and tissue regeneration. However, pure GelMA is limited by the weak mechanical strength and the lack of continuous osteogenic induction environment, which is difficult to meet the needs of bone repair. Moreover, GelMA hydrogels are unable to respond to complex stimuli and therefore are unable to adapt to physiological and pathological microenvironments. This review focused on the functionalization strategies of GelMA hydrogel based bioinks for bone regeneration. The synthesis process of GelMA hydrogel was described in details, and various functional methods to meet the requirements of bone regeneration, including mechanical strength, porosity, vascularization, osteogenic differentiation, and immunoregulation for patient specific repair, etc. In addition, the response strategies of smart GelMA-based bioinks to external physical stimulation and internal pathological microenvironment stimulation, as well as the functionalization strategies of GelMA hydrogel to achieve both disease treatment and bone regeneration in the presence of various common diseases (such as inflammation, infection, tumor) are also briefly reviewed. Finally, we emphasized the current challenges and possible exploration directions of GelMA-based bioinks for bone regeneration.

Glycolysis related lncRNA SNHG3 / miR-139-5p / PKM2 axis promotes castration-resistant prostate cancer (CRPC) development and enzalutamide resistance.

Although androgen deprivation therapy (ADT) by the anti-androgen drug enzalutamide (Enz) may improve the survival level of patients with castration-resistant prostate cancer (CRPC), most patients may eventually fail due to the acquired resistance. The reprogramming of glucose metabolism is one type of the paramount hallmarks of cancers. PKM2 (Pyruvate kinase isozyme typeM2) is a speed-limiting enzyme in the glycolytic mechanism, and has high expression in a variety of cancers. Emerging evidence has unveiled that microRNAs (miRNAs) and long non-coding RNAs (lncRNAs) have impact on tumor development and therapeutic efficacy by regulating PKM2 expression. Herein, we found that lncRNA SNHG3, a highly expressed lncRNA in CRPC via bioinformatics analysis, promoted the invasive ability and the Enz resistance of the PCa cells. KEGG pathway enrichment analysis indicated that glucose metabolic process was tightly correlated with lncRNA SNHG3 level, suggesting lncRNA SNHG3 may affect glucose metabolism. Indeed, glucose uptake and lactate content determinations confirmed that lncRNA SNHG3 promoted the process of glycolysis. Mechanistic dissection demonstrated that lncRNA SNHG3 facilitated the advance of CRPC by adjusting the expression of PKM2. Further explorations unraveled the role of lncRNA SNHG3 as a 'sponge' of miR-139-5p and released its binding with PKM2 mRNA, leading to PKM2 up-regulation. Together, Our studies suggest that lncRNA SNHG3 / miR-139-5p / PKM2 pathway promotes the development of CRPC via regulating glycolysis process and provides valuable insight into a novel therapeutic approach for the disordered disease.

Integrated Insights into Source Apportionment and Source-Specific Health Risks of Potential Pollutants in Urban Park Soils on the Karst Plateau, SW China.

Polycyclic aromatic hydrocarbons (PAHs) and heavy metal(loid)s (HMs) pose risks to environmental and human health. Identification of priority control contaminants is important in guiding the management and control of these synchronous pollutants. A total of 247 soil samples were collected from 64 urban parks in the karst plateau city of Guiyang in SW China to determine the concentrations, spatial distributions, and health risks of PAHs and HMs. The results indicate that dibenz(ah)anthracene and benzo(a)pyrene are the main PAHs species of high ecological risk, and Cr, Mn, and Ni pose elevated ecological risk among the HMs. Four sources were identified for PAHs (biomass burning, coke oven, traffic sources, and coal burning) and HMs (traffic sources, coal burning, industrial sources, and natural sources). The non-carcinogenic risk (NCR) and total carcinogenic risk (TCR) of PAHs were all determined to be negligible and at acceptable levels, several orders of magnitude below those of HMs. The NCR and TCR values of HMs were relatively high, especially for children (11.9% of NCR > 1; 79.1% of TCR > 10-4). Coal burning and natural sources make the greatest contributions to the NCR and TCR values from karst park soils in Guiyang. Considering HMs bioavailability, NCR and TCR values were rather low, due to the high residual HM fractions. Integrated insights into source specific ecological and human health risk indicate future directions for management and control of synchronous PAH and HM pollution, particularly for karst plateau areas. Supplementary Information: The online version contains supplementary material available at 10.1007/s12403-023-00534-3.

Bioprinting EphrinB2-Modified Dental Pulp Stem Cells with Enhanced Osteogenic Capacity for Alveolar Bone Engineering.

Bioprinting, a technology that allows depositing living cells and biomaterials together into a complex tissue architecture with desired pattern, becomes a revolutionary technology for fabrication of engineered constructs. Previously, we have demonstrated that EphrinB2-modified dental pulp stem cells (DPSCs) are expected to be promising seed cells with enhanced osteogenic differentiation capability for alveolar bone regeneration. In this study, we aimed to bioprint EphrinB2-overexpressing DPSCs with low-concentrated Gelatin methacrylate (GelMA) hydrogels into three-dimensional (3D) constructs. The printability of GelMA (5% w/v) and the structural fidelity of bioprinted constructs were examined. Then, viability, proliferation, morphology, and osteogenic differentiation of DPSCs in bioprinted constructs were measured. Finally, the effect of EphrinB2 overexpression on osteogenic differentiation of DPSCs in bioprinted constructs was evaluated. Our results demonstrated that GelMA (5% w/v) in a physical gel form was successfully bioprinted into constructs with various shapes and patterns using optimized printing parameters. Embedded DPSCs showed round-like morphology, and had a high viability (91.93% ± 8.38%) and obvious proliferation (∼1.9-fold increase) 1 day after printing. They also showed excellent osteogenic potential in bioprinted constructs. In bioprinted 3D constructs, EphrinB2-overexpressing DPSCs expressed upregulated osteogenic markers, including ALP, BMP2, RUNX2, and SP7, and generated more mineralized nodules, as compared with Vector-DPSCs. Taken together, this study indicated that fabrication of bioprinted EphrinB2-DPSCs-laden constructs with enhanced osteogenic potential was possible, and 3D bioprinting strategy combined with EphrinB2 gene modification was a promising way to create bioengineered constructs for alveolar bone regeneration.

Combined effects of microplastics and cadmium on the soil-plant system: Phytotoxicity, Cd accumulation and microbial activity.

Microplastics (MPs), an emerging pollutant of concern, widely cooccurred with heavy metals in soil, however, little is known about the combined effects of the interactions of MPs and cadmium (Cd) on the soil-plant system. In this study, the combined effects of several types of MPs and soil Cd contamination on Brassica juncea growth, Cd uptake, and soil microbial carbon metabolism were investigated in a 50-day pot experiment. Aged polyethylene (PE), aged polypropylene (PP), biodegradable polybutylene adipate terephthalate (PBAT) and polylactic acid (PLA) displayed moderate phytotoxicity, with reductions in leaf chlorophyll content and shoot biomass. Compared with the control treatment without MPs or B. juncea, B. juncea growth significantly increased the soil pH by 0.3 pH units, and the growth of B. juncea in the presence of biodegradable PBAT or PLA MPs increased the soil pH by an additional 0.4 or 0.6 pH units, respectively. The presence of PBAT or PLA MPs greatly reduced soil diethylenetriamine pentaacetic acid (DTPA)-extractable Cd concentrations and plant Cd accumulation. The Cd bioconcentration factor was higher in roots than shoots in all treatments except the treatment containing PBAT MPs. The average well color development (AWCD), an indicator of metabolic activity, was highest in the treatment with B. juncea alone and was reduced by both biodegradable and conventional MPs. The microbial utilization efficiency of esters and alcohols was enhanced in the treatment with PBAT MPs, whereas carboxylic acids were preferentially utilized in the treatment with PLA MPs. These findings indicate that co-exposure to MPs and Cd may alter soil microenvironmental characteristics such as soil pH, leading to changes in Cd bioavailability, plant growth and Cd accumulation, and the microbial community's capacity to metabolize carbon. These effects of MPs in soil warrant further exploration.

Comparative Transcriptome Analysis of Pueraria lobata Provides Candidate Genes Involved in Puerarin Biosynthesis and Its Regulation.

Pueraria lobata is a traditional Chinese herb in which an isoflavone C-glucoside, namely puerarin, has received the utmost interest due to its medicinal properties. To date, the biogenesis of puerarin, especially its C-glucosyl reaction in the pathway, remains poorly understood. Moreover, the transcription factors (TFs) that regulate puerarin biosynthesis in P. lobata have not been reported. Here, we performed phytochemical studies on the different developmental stages of the root, stem, and leaf tissues of two P. lobata cultivars, which suggested that both the roots and stems of P. lobata were the sites of puerarin biosynthesis. RNA-sequencing was conducted with the root and stem tissues of the two cultivars under different stages, and the clean reads were mapped to the recently published genome of P. lobata var. thomsonii, yielding the transcriptome dataset. A detailed analysis of the gene expression data, gene coexpression network, and phylogeny proposed several C-GTs that likely participate in puerarin biosynthesis. The first genome-wide analysis of the whole MYB superfamily in P. lobata presented here identified a total of 123 nonredundant PlMYB genes that were significantly expressed in the analyzed tissues. The phylogenetic analysis of PlMYBs with other plant MYB proteins revealed strong PlMYB candidates that may regulate the biosynthesis of isoflavones, such as puerarin.