Biosynthetic Profile in the Co-culture of Aspergillus sydowii and Bacillus subtilis to Produce Novel Benzoic Derivatives.

Microbial co-culture simulates the natural ecosystem through the combination of artificial microbes. This approach has been widely applied in the study of activating silent genes to reveal novel secondary metabolites. However, there are still challenges in determining the biosynthetic pathways. In this study, the effects of microbial co-culture on the morphology of the microbes were verified by the morphological observation. Subsequently, through the strategy combining substrate feeding, stable isotope labeling, and gene expression analysis, the biosynthetic pathways of five benzoic acid derivatives N1-N4 and N7 were demonstrated: the secondary metabolite 10-deoxygerfelin of A. sydowii acted as an inducer to induce B. subtilis to produce benzoic acid, which was further converted into 3-OH-benzoic acid by A. sydowii. Subsequently, A. sydowii used 3-OH-benzoic acid as the substrate to synthesize the new compound N2, and then N1, N3, N4, and N7 were biosynthesized upon the upregulation of hydrolase, hydroxylase, and acyltransferase during co-culture. The plate zone analysis suggested that the biosynthesis of the newly induced compounds N1-N4 was mainly attributed to A. sydowii, and both A. sydowii and B. subtilis were indispensable for the biosynthesis of N7. This study provides an important basis for a better understanding of the interactions among microorganisms, providing new ideas for studying the biosynthetic pathways of the newly induced secondary metabolites in co-culture.

Mulberry Leaf Extract Improves Metabolic Syndrome by Alleviating Lipid Accumulation In Vitro and In Vivo.

Metabolic syndrome (MS) is a metabolic disease with multiple complications. Mulberry leaf extract (MLE) is rich in flavonoids and has great potential in alleviating glucose and lipid metabolism disorders. This study evaluated the effect and mechanism of MLE on the alleviation of MS. The components of the MLE were analyzed, and then the regulation of lipid metabolism by MLE in vitro and in vivo was determined. In a hepatocyte model of oleic acid-induced lipid accumulation, it was found that MLE alleviated lipid accumulation and decreased the expression of genes involved in lipogenesis. Furthermore, MLE improved obesity, insulin resistance, plasma lipid profile, and liver function in MS mice after a 15-week intervention. MLE decreased the expression of SREBP1, ACC, and FAS through the AMPK signaling pathway to inhibit lipid synthesis and increase the level of CPT1A to promote lipid decomposition to achieve its hypolipidemic effect. Meanwhile, MLE was also shown to affect the composition of the gut microbiota and the production of short-chain fatty acids, which contributed to the alleviation of lipid accumulation. Our results suggest that MLE can improve MS by improving lipid metabolism through multiple mechanisms and can be developed into dietary supplements for the improvement of MS.

Immunomodulatory effects of chicken egg yolk antibodies (IgY) against experimental Shewanella marisflavi AP629 infections in sea cucumbers (Apostichopus japonicus).

Skin ulceration syndrome in sea cucumbers is an infectious bacterial disease with fast and high mortality. This study investigated the protection of chicken egg yolk antibodies (IgY) on skin ulcer syndrome in sea cucumbers induced by intraperitoneally injecting Shewanella marisflavi AP629. Inactivated whole S. marisflavi AP629 cells were used as an immunogen to immunize laying hens. The highest titer of the obtained specific IgY by ELISA was 1:90000. Specific IgY significantly inhibited the growth of S. marisflavi AP629 in a liquid medium, dose-dependent manner at concentrations ranging from 0.5 to 2 mg/mL. Results obtained from scanning electron microscopy and confocal laser scanning microscopy showed that specific IgY could make bacteria agglutinate and damage the cell membrane of S. marisflavi AP629, resulting in a decrease of bacterial viability. Sea cucumbers treated with 25, 5, and 1 mg/mL anti-S. marisflavi AP629 IgY could achieve survival rates of 77.5%, 50%, and 22.5% at day 12 when the infection and injection therapy were carried out at the same time, respectively. However, survival rates of sea cucumbers treated with 25 mg/mL of nonspecific IgY were only 7.5% at day 12. All sea cucumbers in the positive control group died within twelve days after bacterial inoculation. Levels of the five humoral immune factors (LYZ, ACP, NOS, SOD, CAT) released by coelomocytes were significantly increased in the specific IgY group compared to the nonspecific IgY and positive control groups within 12 h. However, the activities of LYZ, ACP, and SOD decreased rapidly at the 48 h time point in the specific IgY group, indicating that specific IgY treatment could shorten the time needed to restore balance in sea cucumber immune systems. Oral prophylaxis with egg yolk powders was that all sea cucumbers were challenged with 4.2 × 106 CFU S. marisflavi AP629 by intraperitoneal injection after 60 days of feeding. Survival rates of diets containing 10%, 5%, and 1% specific egg yolk powder were 57.5%, 52.5%, and 30% by day 12, respectively, and the survival rate was 27.5% for the nonspecific group and 22.5% for the positive control group. After feeding for 60 days, enzyme activities of LZY, NOS, and SOD were all significantly enhanced in sea cucumbers fed with specific egg yolk powder when compared to the control group (p < 0.05). This study demonstrated that the phagocytic activities of coelomocytes were significantly stimulated after specific IgY treatment over that of nonspecific IgY or without IgY treatments in sea cucumbers (p < 0.05). Overall, our results revealed that anti-S. marisflavi AP629 IgY has a positive immunomodulatory effect on sea cucumbers infected with S. marisflavi AP629.

Phosphorus Dendrimers Co-deliver Fibronectin and Edaravone for Combined Ischemic Stroke Treatment via Cooperative Modulation of Microglia/Neurons and Vascular Regeneration.

The development of new multi-target combination treatment strategies to tackle ischemic stroke (IS) remains to be challenging. Herein, a proof-of-concept demonstration of an advanced nanomedicine formulation composed of macrophage membrane (MM)-camouflaged phosphorous dendrimer (termed as AK137)/fibronectin (FN) nanocomplexes (NCs) loaded with antioxidant edaravone (EDV) to modulate both microglia and neurons for effective IS therapy is showcased. The created MM@AK137-FN/EDV (M@A-F/E) NCs with a mean size of 260 nm possess good colloidal stability, sustained EDV release kinetics, and desired cytocompatibility. By virtue of MM decoration, the M@A-F/E NCs can cross blood-brain barrier, act on microglia to exert the anti-inflammatory (AK137 and FN) and antioxidative (FN and EDV) effects in vitro for oxidative stress alleviation, microglia M2 polarization, and reduction of pro-inflammatory cytokine secretion, and act on neuron cells to be anti-apoptotic. In a transient middle cerebral artery occlusion rat model, the developed M@A-F/E NCs can exert enhanced antioxidant/anti-inflammatory/anti-apoptotic therapeutic effects to comprehensively regulate the brain microenvironment and promote vascular regeneration to collaboratively restore the blood flow after ischemia-reperfusion. The designed MM-coated NCs composed of all-active ingredients of phosphorous dendrimers, FN, and EDV that can fully regulate the brain inflammatory microenvironment may expand their application scope in other neurodegenerative diseases.

Factors influencing parental fatigue in children with retinoblastoma based on the unpleasant symptoms theory.

Parents of children with retinoblastoma, the most common primary ocular malignant tumour in childhood, bear a heavy caregiving burden and are very susceptible to fatigue. However, little is known about their current status of fatigue and factors influencing fatigue; therefore,this study, based on the theory of unpleasant symptoms, and included 317 parents of children with retinoblastoma in China, whose general demographic data were collected, and on whom the fatigue severity scale (FSS), the generalized anxiety disorder (GAD-7) and 2-item patient health questionnaire (PHQ-2) were administered, from 9 March to 1 June 2020. The parents' FSS score and fatigue incidence were 4.41 ± 1.14 and 67.19%, respectively. The fatigue level was positively correlated with anxiety, depression, education, times of hospitalisations, and treatment types (r = 0.125-0.468, P < 0.05) and negatively correlated with health status, sleep quality, economic status, and family economic situation (r = - 0.120 to - 0.322, P < 0.05). Parent's anxiety level, being an only child and female sex of child influenced parents' fatigue. Healthcare personnel need to focus their attention on this high-risk fatigue group and implement appropriate interventions to reduce their fatigue level, promote their physical and mental health, and facilitate better care for children.

A naphthalimide functionalized fluoran with AIE effect for ratiometric sensing Hg2+ and cell imaging application.

The pollution caused by mercury ions (Hg2+) poses a potential threat to public health. Therefore, monitoring Hg2+ concentration in the environment is necessary and significant. In this work, a naphthalimide functionalized fluoran dye NAF has been prepared, which shows a new red-shift in emission at 550 nm with the maximum intensity in a mixture of water-CH3CN (v/v = 7/3) due to aggregating induced emission (AIE) effect. Meanwhile, NAF can be employed as a Hg2+ ions sensor, which displays a selective and sensitive response to Hg2+ ions by the reduced fluorescence of naphthalimide fluorophore and increased fluorescence of fluoran group, respectively, showing ratiometric fluorescence signal changes with more than 65-fold emission intensity ratio increase and naked eyes visible color change. In addition, the response time is fast (within 1 min) and the sensing can be conducted in a wide pH range (4.0-9.0). Moreover, the detection limit has been evaluated to be 5.5 nM. The sensing mechanism may be attributed to the formation of a π-extended conjugated system due to the Hg2+ ions-induced conversion of spironolactone to the ring-opened form, partially accompanied by the fluorescence resonance energy transfer (FRET) process. Significantly, NAF exhibits suitable cytotoxicity to living HeLa cells, which allows it to be utilized for ratiometric imaging of Hg2+ ions assisted by confocal fluorescence imaging.

Establishment and validation of a risk assessment model for myopia among Chinese primary school students during the COVID-19 pandemic: A lasso regression approach.

Purpose: To construct a risk assessment model for forecasting the likelihood of myopia in elementary school students. Design: A cross-sectional study. Methods: This study utilized convenient sampling and questionnaire survey to collect data from eligible elementary students and their parents during the coronavirus disease 2019 (COVID-19) pandemic period from March to December 2020. The data were divided into training and testing sets in a 7:3 ratio. Lasso regression was employed to screen variables for inclusion in the model to establish a generalized linear model, with a nomogram model as the final result. Results: The study included 1139 elementary students, comprising 54.5 % male and 45.5 % female participants. A total of 37 variables were obtained, which were analyzed using lasso regression. Cross-validation revealed that the best lambda value was 0.04201788. Five variables affecting myopia were identified: three risk and two protective factors. The three risk factors were student age (OR = 1.32), family location (urban vs. rural, OR = 2.33), and parents' occupation (compared with farmer: worker, OR = 2.03; teacher, OR = 1.62; medical worker, OR = 5.64; self-employed, OR = 1.78; civil servant, OR = 1.65; company employee, OR = 1.45; service industries, OR = 3.38; and others, OR = 3.20). The two protective factors were eye distance score (OR = 0.83) and eye health exercise score (OR = 0.95). The model was verified and showed good accuracy with an AUC of 0.778 and Brier score of 0.122 in addition to satisfactory clinical effects. Conclusions: The model effectively predicted the risk of myopia in elementary school students during the COVID-19 pandemic. Using this model, high-risk groups can be identified to provide a foundation for early intervention and follow-up, thereby reducing the incidence of myopia in this population.

Whole-Transcriptome Sequencing Combined with High-Dimensional Proteomic Technologies Reveals the Potential Value of miR-135b-5p as a Biomarker for Hepatocellular Carcinoma.

Purpose: Hepatocellular carcinoma (HCC) is a disease with great heterogeneity and a high mortality rate. It is crucial to identify reliable biomarkers for diagnosis, prognosis, and treatment to improve clinical outcomes in patients with HCC. Alpha-fetoprotein (AFP) is not only a widely used biomarker in clinical practice but also plays a complicated role in HCC, and it has recently been considered to be related to immunotherapy. MicroRNAs (miRNAs) are regarded as key regulators and promising biomarkers of HCC. We investigated the role of an AFP-related miRNA, miR-135b-5p, in HCC progression. Methods: Identification of miR-135b-5p was performed based on a cohort of 65 HCC cases and the liver hepatocellular carcinoma cohort of The Cancer Genome Atlas (Asian people only). A combination of whole-transcriptome sequencing and high-dimensional proteomic technologies was used to study the role of miR-135b-5p in HCC. Results: Upregulation of miR-135b-5p was detected in patients with HCC with high serum AFP levels (AFP > 400 ng/ml). Elevated miR-135b-5p expression was associated with adverse prognosis. We also identified the relevance between high miR-135b-5p expression and tumor-related pathological characteristics, such as Edmondson grade and vascular invasion. We revealed tyrosine kinase nonreceptor 1 as a potential target of miR-135b-5p. Additionally, the transcriptional start site of miR-135b-5p had potential binding sites for SRY-box transcription factor 9, and the stemness properties of tumor cells were more remarkable in HCC with the upregulation of miR-135b-5p. The molecular characterization of the miR-135b-5p-high group was similar to that of the HCC subclasses containing moderately and poorly differentiated tumors. Finally, gene signatures associated with improved clinical outcomes in immune checkpoint inhibitor therapy were upregulated in the miR-135b-5p-high group. Conclusion: miR-135b-5p could be a biomarker for predicting the prognosis and antiprogrammed cell death protein 1 monotherapy response in HCC.

Two-quadrant area structure function analysis for optical surface characterization.

This paper describes the use of the area structure function (SF) for the specification and characterization of optical surfaces. A two-quadrant area SF is introduced because the one-quadrant area SF does not completely describe surfaces with certain asymmetries. Area SF calculations of simulation data and of a diamond turned surface are shown and compared to area power spectral density (PSD) and area autocorrelation function (ACF) representations. The direct relationship between SF, PSD, and ACF for a stationary surface does not apply to non-stationary surfaces typical of optics with figure errors.

A new zinc-organic framework with 1D channel for constructing a ratiometric Al3+-selective sensor and four inputs INHIBIT logic gate.

To develop Al3+ fluorescent sensor is significant because the abnormal levels of Al3+ in environment may pose great threat to human body. Herein, a novel metal-organic framework {Zn(Dpada)(Imdba)·H2O}n (Dpada = 3, 6-di(1H-imidazol-1-yl) pyridazine and Imdba = 2, 2'-iminodibenzoic acid), named Zn-MOF, has been architected with one-dimensional channel under hydrothermal conditions. Zn-MOF exhibits good thermal and solvent stability and can also keep structural integrity over the pH range of 5.0 - 9.0. Fluorescent experiments show that Zn-MOF has high selectivity and sensitivity towards Al3+ via ratiometric fluorescence signal changes (F470 nm/F390 nm) and the detection limit is evaluated to be 0.69 µM. In addition, Zn-MOF performs good recyclability in sensing of Al3+ with at least 5 cycles. Besides, an INHIBIT logic gate has been constructed with chemical ions (Al3+, Cr3+, Fe3+ and Hg2+) as input signals and emission ratio (F470 nm/F390 nm) as output signal. Significantly, Zn-MOF can be applied to tracing Al3+ using real water samples, presenting great potential in water quality monitoring application.

Effect of 1-Deoxynojirimycin on insulin resistance in prediabetic mice based on next-generation sequencing and intestinal microbiota study.

ETHNOPHARMACOLOGICAL RELEVANCE: 1-Deoxynojirimycin (DNJ), the major alkaloid in Morus alba L., is the main effective constituent in "Mulberry twig Alkaloids Tablets" launched in China in 2020. Prediabetes, characterized by insulin resistance, is regarded as the key period for reversing Type 2 diabetes mellitus (T2DM) through lifestyle intervention and glucose-lowering drugs. Besides the excellent activity as an α-glucosidase inhibitor, DNJ also improves insulin sensitivity in T2DM murine models, yet the mechanism is still unclear. Besides, the pharmaceutical effect of DNJ on prediabetes is also undocumented. AIM OF THE STUDY: The aim of this study was to investigate the pharmaceutical effect of DNJ on high-fat and streptozotocin (STZ)-induced prediabetes mice, and to elucidate the mechanism of insulin resistance ameliorated by DNJ. MATERIALS AND METHODS: Oral glucose tolerance test (OGTT) and insulin tolerance test (ITT) were performed to detect blood glucose level and insulin sensitivity in mice. The levels of circulating lipopolysaccharide (LPS), interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α) in the plasma of mice were measured by limulus reagent and enzyme-linked immunosorbent assay (ELISA), respectively. Next-generation sequencing (NGS) and intestinal microbiota sequencing were used to screen the alterations in the transcriptome of liver tissues and to assess the differences in intestinal flora composition, respectively. Expression of cytokine signaling pathway inhibitor 3 (SOCS3), insulin receptor substrate (IRS1), p-IRS1 (Tyr896), occludin, and toll like receptor 4 (TLR4)/NF-κB signaling pathway were confirmed by western blotting. RESULTS: Our study revealed that DNJ decreased the blood glucose level and improve insulin sensitivity in prediabetic mice. DNJ significantly reduced the relative risk of T2DM in prediabetic mice by approximately 83.7%. Mechanistically, DNJ treatment suppressed the circulating levels of LPS, IL-6, and TNF-α in plasma and decreased the inflammatory infiltration in liver and colon tissues. DNJ-treatment increased the abundance of Akkermansia, Bifidobacterium, and Lactobacillus, and decreased the abundance of Enterococcaceae and Lachnospiraceae. Moreover, DNJ suppressed the expression of SOCS3 and the activity of TLR4/NF-κB signaling pathway, meanwhile improving the expression of occludin and the ratio of p-IRS1 (Tyr896)/IRS1. CONCLUSIONS: DNJ effectively ameliorates glucose and lipid metabolism in prediabetic mice, and decreased the relative risk of progression into T2DM from prediabetes. The suppressed immune responses play essential roles in the improvement of insulin resistance by DNJ treatment. In conclusion, DNJ from Morus alba L. is a promising alternative agent in T2DM prevention.

TP53-related signature for predicting prognosis and tumor microenvironment characteristics in bladder cancer: A multi-omics study.

Background: The tumor suppressor gene TP53 is frequently mutated or inactivated in bladder cancer (BLCA), which is implicated in the pathogenesis of tumor. However, the cellular mechanisms of TP53 mutations are complicated, yet well-defined, but their clinical prognostic value in the management of BLCA remains controversial. This study aimed to explore the role of TP53 mutation in regulating the tumor microenvironment (TME), elucidate the effects of TP53 activity on BLCA prognosis and immunotherapy response. Methods: A TP53-related signature based on TP53-induced and TP53-repressed genes was used to construct a TP53 activity-related score and classifier. The abundance of different immune cell types was determined using CIBERSORT to estimate immune cell infiltration. Moreover, the heterogeneity of the tumor immune microenvironment between the high and low TP53 score groups was further evaluated using single-cell mass cytometry (CyTOF) and imaging mass cytometry (IMC). Moreover, pathway enrichment analysis was performed to explore the differential biological functions between tumor epithelial cells with high and low TP53 activity scores. Finally, the receptor-ligand interactions between immune cells and tumor epithelial cells harboring distinct TP53 activity were analyzed by single-cell RNA-sequencing. Results: The TP53 activity-related gene signature differentiated well between TP53 functional retention and inactivation in BLCA. BLCA patients with low TP53 scores had worse survival prognosis, more TP53 mutations, higher grade, and stronger lymph node metastasis than those with high TP53 scores. Additionally, CyTOF and IMC analyses revealed that BLCA patients with low TP53 scores exhibited a potent immunosuppressive TME. Consistently, single-cell sequencing results showed that tumor epithelial cells with low TP53 scores were significantly associated with high cell proliferation and stemness abilities and strongly interacted with immunosuppressive receptor-ligand pairs. Conclusion: BLCA patients with low TP53 scores have a worse prognosis and a more immunosuppressive TME. This TP53 activity-related signature can serve as a potential prognostic signature for predicting the immune response, which may facilitate the development of new strategies for immunotherapy in BLCA.

Integrative Transcriptomic, Lipidomic, and Metabolomic Analysis Reveals Potential Biomarkers of Basal and Luminal Muscle Invasive Bladder Cancer Subtypes.

Muscle invasive bladder cancer (MIBC) is a heterogeneous disease with a high recurrence rate and poor clinical outcomes. Molecular subtype provides a new framework for the study of MIBC heterogeneity. Clinically, MIBC can be classified as basal and luminal subtypes; they display different clinical and pathological characteristics, but the molecular mechanism is still unclear. Lipidomic and metabolomic molecules have recently been considered to play an important role in the genesis and development of tumors, especially as potential biomarkers. Their different expression profiles in basal and luminal subtypes provide clues for the molecular mechanism of basal and luminal subtypes and the discovery of new biomarkers. Herein, we stratified MIBC patients into basal and luminal subtypes using a MIBC classifier based on transcriptome expression profiles. We qualitatively and quantitatively analyzed the lipids and metabolites of basal and luminal MIBC subtypes and identified their differential lipid and metabolite profiles. Our results suggest that free fatty acids (FFAs) and sulfatides (SLs), which are closely associated with immune and stromal cell types, can contribute to the diagnosis of basal and luminal subtypes of MIBC. Moreover, we showed that glycerophosphocholine (GCP)/imidazoles and nucleosides/imidazoles ratios can accurately distinguish the basal and luminal tumors. Overall, by integrating transcriptomic, lipidomic, and metabolomic data, our study reveals specific biomarkers to differentially diagnose basal and luminal MIBC subtypes and may provide a basis for precision therapy of MIBC.

Single-Cell Proteomic Analysis Dissects the Complexity of Tumor Microenvironment in Muscle Invasive Bladder Cancer.

Muscle invasive bladder cancer (MIBC) is a malignancy with considerable heterogeneity. The MIBC tumor microenvironment (TME) is highly complex, comprising diverse phenotypes and spatial architectures. The complexity of the MIBC TME must be characterized to provide potential targets for precision therapy. Herein, an integrated combination of mass cytometry and imaging mass cytometry was used to analyze tumor cells, immune cells, and TME spatial characteristics of 44 MIBC patients. We detected tumor and immune cell clusters with abnormal phenotypes. In particular, we identified a previously overlooked cancer stem-like cell cluster (ALDH+PD-L1+ER-ß-) that was strongly associated with poor prognosis. We elucidated the different spatial architectures of immune cells (excluded, infiltrated, and deserted) and tumor-associated collagens (curved, stretched, directionally distributed, and chaotic) in the MIBC TME. The present study is the first to provide in-depth insight into the complexity of the MIBC TME at the single-cell level. Our results will improve the general understanding of the heterogeneous characteristics of MIBC, potentially facilitating patient stratification and personalized therapy.