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Dysregulated neurite outgrowth and synapse formation underlie many psychiatric disorders, which are also manifested by wolfram syndrome (WS). Whether and how the causative gene WFS1 deficiency affects synapse formation remain elusive. By mirroring human brain development with cerebral organoids, WFS1-deficient cerebral organoids not only recapitulate the neuronal loss in WS patients, but also exhibit significantly impaired synapse formation and function associated with reduced astrocytes. WFS1 deficiency in neurons autonomously delays neuronal differentiation with altered expressions of genes associated with psychiatric disorders, and impairs neurite outgrowth and synapse formation with elevated cytosolic calcium. Intriguingly, WFS1 deficiency in astrocytes decreases the expression of glutamate transporter EAAT2 by NF-κB activation and induces excessive glutamate. When co-cultured with wildtype neurons, WFS1-deficient astrocytes lead to impaired neurite outgrowth and increased cytosolic calcium in neurons. Importantly, disrupted synapse formation and function in WFS1-deficient cerebral organoids and impaired neurite outgrowth affected by WFS1-deficient astrocytes are efficiently reversed with Riluzole treatment, by restoring EAAT2 expression in astrocytes. Furthermore, Riluzole rescues the depressive-like behavior in the forced swimming test and the impaired recognition and spatial memory in the novel object test and water maze test in Wfs1 conditional knockout mice. Altogether, our study provides novel insights into how WFS1 deficiency affects synapse formation and function, and offers a strategy to treat this disease.

Surgical patients with depleted skeletal muscle mass tend to have a worse outcome. Whether perioperative change of urea to creatinine ratio (CUCR) can reflect muscle wasting and predict postoperative complications have not been investigated. This study aimed to evaluate the relationship of perioperative CUCR with postoperative complications and skeletal muscle wasting in pancreatic cancer patients undergoing pancreatoduodenectomy (PD).Pancreatic cancer patients undergoing PD were included retrospectively. The association between postoperative complications and perioperative CUCR as well as other nutritional biomarkers was analyzed. In a subset of patients with serial CT scans, the correlation of the CUCR and the changes of CT-derived skeletal muscle area (SMA) were tested. Furthermore, the capacity of complication prediction of CUCR and CT-derived parameter were compared in these patients.A total of 321 surgical patients were included. Univariable and multivariable logistic regression demonstrated CUCR was a strong predictor for complications in these patients, independent of age, BMI and comorbidity. Patients with CUCR above the median have higher complication rate (p=0.007) and longer postoperative days to discharge (p=0.017). In a subset patients with both pre- and postoperative digital abdominal CT scans, spearman correlation analysis shown both L3 muscle area and L4-psoas area were significantly correlated with CUCR (R2=0.64, p0.05; R2=0.62, p0.05, respectively).Perioperative CUCR is an independent predictor for postoperative complications in pancreatic cancer patients undergoing PD. Elevated CUCR is a reflection of skeletal muscle wasting in postoperative surgical patients.

Mouse Nsmce1 gene is the homolog of non-structural maintenance of chromosomes element 1 (NSE1) that is mainly involved in maintenance of genome integrity, DNA damage response, and DNA repair. Defective DNA repair may cause neurological disorders such as Alzheimer's disease (AD). So far, there is no direct evidence for the correlation between Nsmce1 and AD. In order to explore the function of Nsmce1 in the regulation of nervous system, we have overexpressed or knocked down Nsmce1 in the mouse hippocampal neuronal cells (MHNCs) HT-22 and detected its regulation of AD marker genes as well as cell proliferation. The results showed that the expression of App, Bace2, and Mapt could be inhibited by Nsmce1 overexpression and activated by the knockdown of Nsmce1. Moreover, the HT-22 cell proliferation ability could be promoted by Nsmce1 overexpression and inhibited by knockdown of Nsmce1. Furthermore, we performed a transcriptomics study by RNA sequencing (RNA-seq) to evaluate and quantify the gene expression profiles in response to the overexpression of Nsmce1 in HT-22 cells. As a result, 224 significantly dysregulated genes including 83 upregulated and 141 downregulated genes were identified by the comparison of Nsmce1 /+ to WT cells, which were significantly enriched in several Gene Ontology (GO) terms and pathways. In addition, the complexity of the alternative splicing (AS) landscape was increased by Nsmce1 overexpression in HT-22 cells. Thousands of AS events were identified to be mainly involved in the pathway of ubiquitin-mediated proteolysis (UMP) as well as 3 neurodegenerative diseases including AD. The protein-protein interaction network was reconstructed to show top 10 essential genes regulated by Nsmce1. Our sequencing data is available in the Gene Expression Omnibus (GEO) database with accession number as GSE113436. These results may provide some evidence of molecular and cellular functions of Nsmce1 in MHNCs.

The occurrence of microplastics (MPs) in the environment has been gaining widespread attention globally. MP-colonizing microorganisms are important links for MPs contamination in various ecosystems, but have not been well understood. To partially address this issue, the present study investigated biofilm formation by microorganisms originating from lake water on low-density polyethylene (LDPE) MPs using a cultivation approach and the surface-related effects on the MP-associated microbial communities using 16S rRNA high-throughput sequencing. With the addition of nonionic surfactants and UV-irradiation pretreatment that changed the surface properties of LDPE MPs, more microorganisms were colonized on LDPE surface. Microbial community analysis indicated that LDPE MPs were primarily colonized by the phyla Proteobacteria, Bacteroidetes and Firmicutes, and the surface roughness and hydrophobicity of MP were important factors shaping the LDPE MP-associated microbial community structure. Half of the top 20 most abundant genera colonizing on LDPE were found to be potential pathogens, e.g., plant pathogens Agrobacterium, nosocomial pathogens Chryseobacterium and fish pathogens Flavobacterium. This study demonstrated rapid bacterial colonization of LDPE MPs in lake water microcosms, the role of MPs as transfer vectors for harmful microorganisms in lake water, and provided a first glimpse into the effect of surface properties on LDPE MP-associated biofilm communities.

Although many of the genetic loci associated with breast cancer risk have been reported, there is a lack of systematic analysis of regulatory networks composed of different miRNAs and mRNAs on survival analysis in breast cancer. To reconstruct the microRNAs-genes regulatory network in breast cancer, we employed the expression data from The Cancer Genome Atlas (TCGA) related to five essential miRNAs including miR-21, miR-22, miR-210, miR-221, and miR-222, and their associated functional genomics data from the GEO database. Then, we performed an integration analysis to identify the essential target factors and interactions for the next survival analysis in breast cancer. Based on the results of our integrated analysis, we have identified significant common regulatory signatures including differentially expressed genes, enriched pathways, and transcriptional regulation such as interferon regulatory factors (IRFs) and signal transducer and activator of transcription 1 (STAT1). Finally, a reconstructed regulatory network of five miRNAs and 34 target factors was established and then applied to survival analysis in breast cancer. When we used expression data for individual miRNAs, only miR-21 and miR-22 were significantly associated with a survival change. However, we identified 45 significant miRNA-gene pairs that predict overall survival in breast cancer out of 170 one-on-one interactions in our reconstructed network covering all of five miRNAs, and several essential factors such as PSMB9, HLA-C, RARRES3, UBE2L6, and NMI. In our study, we reconstructed regulatory network of five essential microRNAs for survival analysis in breast cancer by integrating miRNA and mRNA expression datasets. These results may provide new insights into regulatory network-based precision medicine for breast cancer.

Riverine runoff is a significant transport pathway for microplastics (MPs) discharged from land-based sources to marine environments where MPs accumulate. Knowledge of riverine MP-associated biofilms will improve the understanding of the fate and potential effects of MPs in marine environments. This study aimed to characterize the microbial biofilms colonizing MPs in the riverine water of the Pearl River Delta, China, and identify the seasonal, geographical and environmental influences on MP-associated communities. We sampled MPs and the surrounding surface water from eight outlets in three seasons and analyzed their microbial communities by Illumina sequencing of the 16S rRNA gene libraries. Across all sampling seasons and locations, abundant MP-colonizing taxa belonged to the phylum Proteobacteria, which suggested initial biofilm development on those MPs. The structure and composition of MP-attached microbial communities varied with respect to season and location, and the microbial diversity of the MP-associated biofilm communities decreased in June compared with that in the April and November sampling events. Opportunistic pathogens of the genus Acinetobacter were significantly enriched on the MP surfaces for all sampling events. Among the 15 environmental variables examined, the main drivers of MP-associated biofilm community composition included IC, alkalinity, TOC, TDS, Cl-, NO3-, NO2- and pH. This study provides an insight into the environmental factors that shape microbial biofilm colonization on MPs in estuary environments and a further understanding of the structure, diversity and ecological roles of MP-associated communities.

BackgroundSurgical Patients with depleted skeletal muscle mass tend to have a worse outcome in the perioperative period, but the muscle mass is difficult to quantify. Here, we aimed to evaluate the relationship of perioperative change of serum urea to creatinine ratio (CUCR) with postoperative complication and skeletal muscle mass in pancreatic cancer patients underwent pancreatoduodenectomy (PD).MethodsPancreatic cancer patients underwent PD were included retrospectively. We analyzed the association of serum urea, creatinine, and urea to creatinine ratio (UCR) between patients with and without major postoperative complications. In a subset of patients with serial CT scans, we tested the correlation of the CUCR and the changes of CT-derived skeletal muscle area (SMA). Furthermore, the capacity of complication prediction of CUCR and CT-derived parameter were compared in these patients.ResultsWe included 321 surgical patients from January 2014 to July 2020. Patients were grouped with high- and low CUCR according ROC curve (cut-off: 71.3 for male and 67.4 for female). Patients with high CUCR (n=175) have higher complication rate (30.3% vs. 21.1%, p=0.002), including POPF (23.4% vs. 15.1%, p=0.017), infection rate (16.0% vs. 11.0%, p=0.043) and longer postoperative days to discharge (16.7±6.5, vs. 12.1±4.2, p=0.031). Multivariable logistic regression demonstrated CUCR was an independent predictor for complications in these patients, independent of age, sex and BMI. In a subset patients with serial CT scans, spearman correlation analysis shown both L3 muscle area and L4-psoas area were significantly correlated with CUCR (R2 = 0.64, p < 0.05; R2 = 0.62, p < 0.05, respectively). ConclusionPerioperative CUCR is an independent predictor for postoperative complications in pancreatic cancer patients underwent PD procedure. Elevated CUCR is a reflection of skeletal muscle wasting in postoperative surgical patients.

BackgroundPostoperative complications remain high in patients underwent pancreaticoduodenectomy (PD). Body composition has been shown closely related to the outcome in critical and surgical patients. The aim of the present study was to investigate whether preoperative skeletal muscle condition and postoperative nutrition would affect major complications in patients underwent PD.MethodsThis retrospective study included 265 patients underwent PD with a CT scan of the abdomen within 1 week before surgery. Body compartment data was extracted from the L3 level of the CT scan, which included skeletal muscle area and density, subcutaneous and visceral fat area. Univariable and multivariable regression analyses were performed to investigate correlations between the above variables and postoperative complications. Furthermore, a subgroup analysis was conducted to explore the relationship between postoperative nutrition strategy and the outcome.ResultsOf all the 265 patients, the mean age and BMI was 59.5±13.9 years and 23.1 ±3.6 kg/m2, respectively. Major complications occurred in 81 patients (30.6%). Cutoff values for skeletal muscle depletion were defined by ROC curve analysis from postoperative complications in skeletal muscle index (SMI) (male 47.32 cm2/m2 and female 40.65 cm2/m2). Univariable analysis showed postoperative complications were significantly associated with age, sex, systemic comorbidity, SMI, skeletal muscle density (SMD) and visceral adipose tissue index (VATI). Multivariable regression reveled only age (OR 1.49, 95% CI 1.22-1.83, p=0.026), SMI (OR 0.77, 95%CI 0.51-0.94, p=0.015) and SMD (OR 0.85, 95%CI 0.64-1.03, p=0.029) were independent predictors for major complications. Subgroup analysis showed the initial parenteral nutrition time (IPNT) (OR1.89, 95% CI 1.43-2.49, p=0.032) and average protein delivery (APD) (OR 0.76, 95% CI 0.53-0.89, p=0.021) were significantly associated with major complications in patients with lower SMI. However, in normal SMI patients, such association was not significant enough.ConclusionsPreoperative skeletal muscle index and density were independently associated with major complications in patients underwent PD. In patients with lower SMI, early parenteral nutrition and higher protein delivery were related to better outcome.

Purpose Fungal keratitis (FK) is an eye disease that can lead to blindness and has a high incidence worldwide. At present, there is no effective treatment for this disease. There are innate immune response mechanisms that protect against fungal infections. One example is C-type lectin receptors (CLRs), which can identify fungal invaders and trigger signal transduction pathways and cellular responses to eliminate pathogens. However, previous studies have focused mostly on single-receptor factors, and a systematic analysis of the genetic factors underlying the pathogenesis of FK has not been conducted. This study aimed to investigate the molecular mechanisms of FK in terms of genomics and to further elucidate its pathogenesis. Methods We performed a transcriptome analysis of a mouse model of FK using RNA sequencing to obtain the relevant gene expression profiles and to identify differentially expressed genes, signaling pathways, and regulatory networks of the key genetic factors in the pathogenesis of murine FK. Results Several genes that are significantly associated with FK and serve as markers of FK, such as the inflammatory cytokine genes IL1B, IL6, IL10, IL23, and TNF, were identified. The mRNA and protein expression patterns of IL-1β, IL-6, and TNF-α in the corneas of mice with FK were validated by quantitative RT-PCR and Luminex multiplex assay technology. The Wnt, cGMP-PKG, and Hippo signaling pathways were significantly enriched during fungal infection of mouse corneas. Conclusions Our study may help to elucidate the mechanisms of FK pathogenesis and to identify additional candidate drug targets for the treatment of FK.

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To study the cellular and molecular function of peroxisome proliferator-activated receptor γ (PPARγ) in skeletal muscle differentiation, we have generated inducible gain-of-function to overexpress PPARγ in C2C12 myoblasts. In order to identify PPARγ targets, RNA sequencing (RNA-seq) was used to evaluate and quantify the transcriptomes and expression patterns during myogenic differentiation under the overexpression of PPARγ. The formation of myotubes and the expression of muscle-specific myogenic genes such as MyoD and MyoG may be inhibited by PPARγ overexpression. Multiple genes and pathways were significantly involved in this process, including 11 genes such as Fndc9 and Slc14a1 with fundamental change of regulation modes, 9 genes of which were validated by the data of qRT-PCR. Our studies demonstrate that PPARγ would play critical roles on myoblasts differentiation, mediating crosstalk among several pathways and transcription factors. Our data is available in the Gene Expression Omnibus (GEO) database with the accession number as GSE99399.

Combination with genomic DNA is one of the important ways for microRNAs (miRNAs) to perform biological processes. However, because of lack of an experimental method, the identified genomic sites targeted by microRNA were only located in the promoter and enhancer regions. In this study, based on affinity purification of labeled biotin at the 3′-end of miRNAs, we established an efficiently experimental method to screen miRNA binding sequences in the whole genomic regions in vivo. Biotinylated miR-373 was used to test our approach in MCF-7 cells, and then Sanger and next-generation sequencing were used to screen miR-373 binding sequences. Our results demonstrated that the genomic fragments precipitated by miR-373 were located not only in promoter but also in intron, exon, and intergenic. Eleven potentially miR-373 targeting genes were selected for further study, and all of these genes were significantly regulated by miR-373. Furthermore, the targeting sequences located in E-cadherin, cold-shock domain-containing protein C2 (CSDC2), and PDE4D genes could interact with miR-373 in MCF-7 cells rather than HeLa cells, which is consistent with our data that these three genes can be regulated by miR-373 in MCF-7 cells while not in HeLa cells. On the whole, this is an efficient method to identify miRNA targeting sequences in the whole genome.

Di-(2-ethylhexyl) phthalate (DEHP) is a widely used plasticizer. It has neurotoxicity and exposure to it causes impairment of neurodevelopment, behavior and cognition. However, the molecular mechanisms responsible for the DEHP-induced neurotoxicity are not yet clearly defined. Tumor necrosis factor-induced protein 1 (TNFAIP1) was first discovered in umbilical vein endothelial cells and was further found to be important in the progress of Alzheimer's disease. Herein we explore the mechanism of TNFAIP1 in DEHP-induced neurotoxicity with the involvement of cyclic AMP response elements binding protein (CREB) signaling pathway in a mouse neuroblastoma cell line (N2a cells). We found that exposure to DEHP induced apoptosis and downregulated the expression of brain-derived neurotrophic factor (BDNF), synaptic proteins PSD 95 and synapsin-1 while upregulated the expression of TNFAIP1 and decreased the levels of phosphorylated Akt, CaMK Ⅳ, catalytic subunits of PKA and CREB in CREB signaling pathway. Knockdown of TNFAIP1 using TNFAIP1 small interfering RNA (siRNA) expression vector prevented DEHP from inhibiting CREB pathway, thus reduced apoptosis and restored expression of BDNF, PSD 95 and synapsin-1. Our data indicate that downregulation of TNFAIP1 prevents DEHP-induced neurotoxicity via activating CREB pathway. Therefore, TNFAIP1 is a potential target for relieving the DEHP-induced neurotoxicity and related neurological disorders.

Evidence from biochemical liver function index and histopathology analysis suggested that selenium could effectively repair the liver injury caused by beta-cypermethrin (β-CYP). However, the molecular mechanism of selenium against liver injury induced by β-CYP remains unclear. In the present study, dynamic changes in gene expression profiles before and after the treatment of Na2SeO3 in liver injury mice were analyzed by using RNA sequencing. As a result, several essential genes and pathways were identified to be significantly associated with this process. In particular, ten genes including Cyp2j11, Cyp2b10, Cyp3a13, Dhrs9, Socs2, Stat4, Gm13305, Cyp3a44, Retsat, and Cyp26b1 were significantly enriched in the functional categories related to retinol metabolism, linoleic acid metabolism, and Jak-STAT signaling pathway. Among them, the expression patterns of nine genes were validated by qRT-PCR, except for Cyp3a44. Furthermore, we have constructed the associated regulatory network based on the identified targets revealed by high throughput screening. Our study may provide insight into the molecular mechanism underlying the protective effect of selenium against liver injury induced by β-CYP in mammals.

Activation of signal transducer and activator of transcription 3 (STAT3) by leukemia inhibitory factor (LIF) maintains mouse embryonic stem cell (mESC) self-renewal. Our previous study showed that trans-acting transcription factor 5 (Sp5), an LIF/STAT3 downstream target, supports mESC self-renewal. However, the mechanism by which Sp5 exerts these effects remains elusive. Here, we found that Nanog is a direct target of Sp5 and mediates the self-renewal-promoting effect of Sp5 in mESCs. Overexpression of Sp5 induced Nanog expression, while knockdown or knockout of Sp5 decreased the Nanog level. Moreover, chromatin immunoprecipitation (ChIP) assays showed that Sp5 directly bound to the Nanog promoter. Functional studies revealed that knockdown of Nanog eliminated the mESC self-renewal-promoting ability of Sp5. Finally, we demonstrated that the self-renewal-promoting function of Sp5 was largely dependent on its zinc finger domains. Taken together, our study provides unrecognized functions of Sp5 in mESCs and will expand our current understanding of the regulation of mESC pluripotency.