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.

An immunogenic cell death-related classification predicts prognosis and response to immunotherapy in kidney renal clear cell carcinoma.

Introduction: Immunogenic cell death (ICD) is a form of regulated cell death that activates an adaptive immune response in an immunocompetent host and is particularly sensitive to antigens from tumor cells. Kidney clear cell carcinoma (KIRC) is an immunogenic tumor with extensive tumor heterogeneity. However, no reliable predictive biomarkers have been identified to reflect the immune microenvironment and therapeutic response of KIRC. Methods: Therefore, we used the CIBERSORT and ESTIMATE algorithms to define three ICD clusters based on the expression of ICD-related genes in 661 KIRC patients. Subsequently, we identified three different ICD gene clusters based on the overlap of differentially expressed genes (DEGs) within the ICD clusters. In addition, principal component analysis (PCA) was performed to calculate the ICD scores. Results: The results showed that patients with reduced ICD scores had a poorer prognosis and reduced transcript levels of immune checkpoint genes regulated with T cell differentiation. Furthermore, the ICD score was negatively correlated with the tumor mutation burden (TMB) value of KICD. patients with higher ICD scores showed clinical benefits and advantages of immunotherapy, indicating that the ICD score is an accurate and valid predictor to assess the effect of immunotherapy. Discussion: Overall, our study presents a comprehensive KICD immune-related ICD landscape that can provide guidance for current immunotherapy and predict patient prognosis to help physicians make judgments about the patient's disease and treatment modalities, and can guide current research on immunotherapy strategies for KICD.

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.

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.

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.

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.

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.

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.

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.

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.

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.

Alternative ANKHD1 transcript promotes proliferation and inhibits migration in uterine corpus endometrial carcinoma.

Alternative splicing (AS) is common in gene expression, and abnormal splicing often results in several cancers. Overall survival-associated splicing events (OS-SEs) have been used to predict prognosis in cancer. The aim of this study was to investigate the presence and function of OS-SEs in uterine corpus endometrial carcinoma (UCEC). Based on TCGA and TCGASpliceSeq databases, gene expression and the AS data of UCEC samples were retrieved. An alternate terminator of ANKHD1 transcripts named ANKHD1-BP3 was found to be significantly related to metastasis and OS in UCEC and significantly associated with HSPB1. The upregulated expression of HSPB1 induced downregulation of ANKHD1-BP3 and promoted tumor metastasis. These findings indicate that HSPB1, a splicing factor, regulates the expression of ANKHD1-BP3 to promote metastasis in UCEC.

Zn3V4(PO4)6: A New Rocking-Chair-Type Cathode Material with High Specific Capacity Derived from Zn2+/H+ Cointercalation for Aqueous Zn-Ion Batteries.

Phosphate cathode materials with a stable and open framework structure are expected to be one of the favorable cathode materials for aqueous zinc-ion batteries (AZIBs). However, the slow migration rate of Zn2+ and complex mechanism in aqueous electrolyte are serious problems that limit their application at the present. Here, a new rocking-chair-type cathode material Zn3V4(PO4)6@C (ZVP@C) for AZIBs is synthesized for the first time and evaluated using a composite carbon coating to improve the electronic conductivity. Benefiting from the two-electron reaction of vanadium and the cointercalation of Zn2+/H+, ZVP@C/30%BP delivers a specific capacity as high as 120 mAh·g-1 at 0.04 A·g-1. A good capacity retention of 80% after 400 cycles at 1 A·g-1 is also obtained, which is attributed to the stable crystal structure and the cointercalation reaction of Zn2+/H+. The reaction mechanism is investigated by in situ X-ray diffraction (XRD), ex situ XRD, ex situ X-ray photoelectron spectroscopy (XPS), and energy dispersive spectroscopy (EDS). This work not only provides a new phosphate cathode material for AZIBs but also gives a new strategy for improving the specific capacity of phosphate cathode material.

The Progress of Social Isolation in Patients with Enterostomy during Discharge Transition Period.

In terms of social and psychological health of enterostomy patients during hospitalization and discharge transition period, the degree of social isolation in patients during discharge transition period is higher than that during hospitalization period, which is usually manifested by poor self-perception of body image changes. Self-esteem (shame) frustration, severe negative emotions, low psychosocial adjustment, and other factors are closely related to postoperative complications, coping self-efficacy, social support level, family living conditions, and other influencing factors. This is an important reason why patients are difficult to adapt to ostomy life and low quality of life. At present, it is believed that the social isolation related evaluation scale can be used to understand the status quo of patients with social isolation and provide nursing guidance, so as to better cope with the adverse medical outcomes caused by social isolation.

Association of eHealth Literacy with Health Promotion Behaviors of Community-Dwelling Older People: The Chain Mediating Role of Self-Efficacy and Self-Care Ability.

In the digital age, electronic health literacy (eHealth literacy) of community-dwelling older people plays a potentially important role in their health behaviors which are critical for health outcomes. Researchers have documented that self-efficacy and self-care ability are related to this relationship. This study aimed to assess the relationship between eHealth literacy and health promotion behaviors among older people living in communities and explore the chain mediating role of self-efficacy and self-care ability. For this cross-sectional study, we used data from 425 older adults at 3 communities in Qingdao, Shandong Province in Northeastern China, from June to September 2021. Path analysis using the structural equation model was performed. We found that eHealth literacy was significantly associated with health promotion behaviors in older people. Additionally, eHealth literacy indirectly affected health promotion behaviors through self-efficacy and self-care ability, respectively. In addition, the chain mediation effect was identified in the relationship of eHealth literacy and health promotion behaviors: eHealth literacy→ self-efficacy→ self-care ability→ health promotion behaviors. These findings offer promising directions for developing interventions to modify older adults' health behaviors through enhancing their eHealth literacy. These interventions should integrate components that target improving the self-efficacy and self-care ability of older people.

B3GNT3 as a prognostic biomarker and correlation with immune cell infiltration in lung adenocarcinoma.

Background: Lung adenocarcinoma (LUAD) is the most common malignant cancer in humans and because of low long-term survival rates, exploration of the molecular mechanisms underlying its progression, as well as novel prognostic predictors, is urgently needed. B3GNT3, a type II transmembrane protein located in the Golgi apparatus, is essential for forming extended core 1 oligosaccharides and is reportedly involved in malignant transformation. Methods: The Cancer Genome Atlas (TCGA) and GSE68465 were used to analyze the expression of B3GNT3 in LUAD and normal tissues and overall survival. Real time quantitative polymerase chain reaction (qPCR) and western blot were conducted to measure the mRNA and protein levels of B3GNT3, respectively. Functional enrichment of differentially expressed genes was explored using Gene Ontology and Kyoto Encyclopedia of Genes and Genomes pathway analyses. We performed univariate and multivariate Cox regression analyses and a meta-analysis to reveal an independent factor for LUAD. We evaluated the correlation between immune infiltration levels and cumulative survival in the TIMER database. The correlation between B3GNT3 and immune cell infiltration was assessed via Cell-type Identification by Estimating Relative Subsets of RNA Transcripts (CIBERSORT). The association of DNA methylation of B3GNT3 and prognosis was determined. A nomogram that incorporated expression and clinical features was additionally built for prognostic prediction. Cell proliferation, cloning, and invasion were conducted to validate the roles of B3GNT3 in LUAD. Results: B3GNT3 was more highly expressed in LUAD tissues than in normal lung tissues, consistent with the mRNA and protein levels in LUAD cells. B3GNT3 was an independent factor for LUAD. Moreover, the levels of B3GNT3 were related to immune cell infiltration in LUAD microenvironments. DNA methylation of B3GNT3 correlated with the mRNA of B3GNT and overall survival of LUAD patients. The expression of B3GNT3 was highly valuable for the prediction of diagnosis. Knockdown of B3GNT3 inhibited LUAD cell viability and cloning ability, and hindered invasion. Conclusions: B3GNT3 was highly associated with immune cell infiltration, acting as an important biomarker for the prognosis and diagnosis of LUAD.

Functional redundancy among Polycomb complexes in maintaining the pluripotent state of embryonic stem cells.

Polycomb group proteins assemble into multi-protein complexes, known as Polycomb repressive complexes 1 and 2 (PRC1 and PRC2), that guide cell fate decisions during embryonic development. PRC1 forms an array of biochemically distinct canonical PRC1 (cPRC1) or non-canonical PRC1 (ncPRC1) complexes characterized by the mutually exclusive presence of PCGF (PCGF1-PCGF6) paralog subunit; however, whether each one of these subcomplexes fulfills a distinct role remains largely controversial. Here, by performing a CRISPR-based loss-of-function screen in embryonic stem cells (ESCs), we uncovered a previously unappreciated functional redundancy among PRC1 subcomplexes. Disruption of ncPRC1, but not cPRC1, displayed severe defects in ESC pluripotency. Remarkably, coablation of non-canonical and canonical PRC1 in ESCs resulted in exacerbation of the phenotype observed in the non-canonical PRC1-null ESCs, highlighting the importance of functional redundancy among PRC1 subcomplexes. Together, our studies demonstrate that PRC1 subcomplexes act redundantly to silence lineage-specific genes and ensure robust maintenance of ESC identity.

RNAi of Sterol Δ24-Isomerase Implicated Its Involvement in Physalin Biosynthesis in Physalis angulata L.

Physalis angulata is a renowned traditional Chinese medicine for the treatment of various conditions. Physalin is the major type of bioactive constituents conferring medicinal properties of P. angulata. Despite the medicinal importance, the pathways leading to physalin are largely unknown. In this study, we employed a transcriptomic approach to identify a Pa24ISO gene from P. angulata. Through heterologous expression in yeast, Pa24ISO was revealed to catalyze an isomerization reaction in converting 24-methylenecholesterol to 24-methyldesmosterol. Real-time PCR analysis showed that the abundance of Pa24ISO transcripts correlated with the accumulation pattern of physalin B in different tissues of P. angulata. A direct role of Pa24ISO in channeling of 24-methylenecholesterol for physalin B biosynthesis was illustrated by suppressing the gene in P. angulata via the VIGS approach. Down-regulation of Pa24ISO led to reduced levels of 24-methyldesmosterol and physalin B, accompanied with an increase of campesterol content in P. angulata. The results supported that 24ISO is involved in physalin biosynthesis in plants.