Uncovering global-scale risks from commercial chemicals in air.

Commercial chemicals are used extensively across urban centres worldwide1, posing a potential exposure risk to 4.2 billion people2. Harmful chemicals are often assessed on the basis of their environmental persistence, accumulation in biological organisms and toxic properties, under international and national initiatives such as the Stockholm Convention3. However, existing regulatory frameworks rely largely upon knowledge of the properties of the parent chemicals, with minimal consideration given to the products of their transformation in the atmosphere. This is mainly due to a dearth of experimental data, as identifying transformation products in complex mixtures of airborne chemicals is an immense analytical challenge4. Here we develop a new framework-combining laboratory and field experiments, advanced techniques for screening suspect chemicals, and in silico modelling-to assess the risks of airborne chemicals, while accounting for atmospheric chemical reactions. By applying this framework to organophosphate flame retardants, as representative chemicals of emerging concern5, we find that their transformation products are globally distributed across 18 megacities, representing a previously unrecognized exposure risk for the world's urban populations. More importantly, individual transformation products can be more toxic and up to an order-of-magnitude more persistent than the parent chemicals, such that the overall risks associated with the mixture of transformation products are also higher than those of the parent flame retardants. Together our results highlight the need to consider atmospheric transformations when assessing the risks of commercial chemicals.

APLN promotes the proliferation, migration, and glycolysis of cervical cancer through the PI3K/AKT/mTOR pathway.

Apelin (APLN) is an endogenous ligand of the G protein-coupled receptor APJ (APLNR). APLN has been implicated in the development of multiple tumours. Herein, we determined the effect of APLN on the biological behaviour and underlying mechanisms of cervical cancer. The expression and survival curves of APLN were determined using Gene Expression Profiling Interactive Analysis. The cellular functions of APLN were detected using CCK-8, clone formation, EdU, Transwell assays, flow cytometry, and seahorse metabolic analysis. The underlying mechanisms were elucidated using gene set enrichment analysis and Western blotting. APLN was upregulated in the samples of patients with cervical cancer and is associated with poor prognosis. APLN knockdown decreased the proliferation, migration, and glycolysis of cervical cancer cells. The opposite results were observed when APLN was overexpressed. Mechanistically, we determined that APLN was critical for activating the PI3K/AKT/mTOR pathway via APLNR. APLN receptor inhibitor ML221 reversed the effect of APLN overexpression on cervical cancer cells. Treatment with LY294002, the PI3K inhibitor, drastically reversed the oncological behaviour of APLN-overexpressing C-33A cells. APLN promoted the proliferation, migration, and glycolysis of cervical cancer cells via the PI3K/AKT/mTOR pathway.

Development of multiplex allele-specific RT-qPCR assays for differentiation of SARS-CoV-2 Omicron subvariants.

Quick differentiation of current circulating variants and the emerging recombinant variants of SARS-CoV-2 is essential to monitor their transmissions. However, the widely applied gene sequencing method is time-consuming and costly especially when facing recombinant variants, because a large part or whole genome sequencing is required. Allele-specific reverse transcriptase real time RT-PCR (RT-qPCR) represents a quick and cost-effective method for SNP (single nucleotide polymorphism) genotyping and has been successfully applied for SARS-CoV-2 variant screening. In the present study, we developed a panel of 5 multiplex allele-specific RT-qPCR assays targeting 20 key mutations for quick differentiation of the Omicron subvariants (BA.1 to BA.5 and their descendants) and the recombinant variants (XBB.1 and XBB.1.5). Two parallel multiplex RT-qPCR reactions were designed to separately target the prototype allele and the mutated allele of each mutation in the allele-specific RT-qPCR assay. Optimal annealing temperatures, primer and probe dosage, and time for annealing/extension for each reaction were determined by multi-factor and multi-level orthogonal test. The variation of Cp (crossing point) values (ΔCp) between the two multiplex RT-qPCR reactions was applied to determine if a mutation occurs or not. SARS-CoV-2 subvariants and related recombinant variants were differentiated by their unique mutation patterns. The developed multiplex allele-specific RT-qPCR assays exhibited excellent analytical sensitivities (with limits of detection (LoDs) of 1.47-18.52 copies per reaction), wide linear detection ranges (109-100 copies per reaction), good amplification efficiencies (88.25 to 110.68%), excellent reproducibility (coefficient of variations (CVs) < 5% in both intra-assay and inter-assay tests), and good clinical performances (99.5-100% consistencies with Sanger sequencing). The developed multiplex allele-specific RT-qPCR assays in the present study provide an alternative tool for quick differentiation of the SARS-CoV-2 Omicron subvariants and their recombinant variants. KEY POINTS: • A panel of five multiplex allele-specific RT-qPCR assays for quick differentiation of 11 SARS-CoV-2 Omicron subvariants (BA.1, BA.2, BA.4, BA.5, and their descendants) and 2 recombinant variants (XBB.1 and XBB.1.5). • The developed assays exhibited good analytical sensitivities and reproducibility, wide linear detection ranges, and good clinical performances, providing an alternative tool for quick differentiation of the SARS-CoV-2 Omicron subvariants and their recombinant variants.

Triclosan is the Predominant Antibacterial Compound in Ontario Sewage Sludge.

Sewage treatment plants (STPs) accumulate both antibiotic and nonantibiotic antimicrobial compounds that can select for antibiotic resistant bacteria. Herein, we aimed to identify the predominant antibacterial compounds impacting E. coli from Ontario sewage sludge consisting of thousands of unknown compounds. Among the 10 extracted sludge samples, 6 extracts exerted significant growth inhibition effects in E. coli. A total of 103 compounds were tentatively detected across the 10 sludge samples by suspect screening, among which the bacterial enoyl-ACP reductase (FabI) inhibitor triclocarban was detected at the highest abundance. A hypomorphic FabI knockdown E. coli strain was highly susceptible to the sludge extracts, confirming FabI inhibitors as the primary antibacterial compounds in the sludge. Protein affinity pulldown identified triclosan as the major ligand binding to a His-tagged FabI protein from the sludge, despite the higher abundance of triclocarban in the same samples. Effect-directed analysis was used to determine the contributions of triclosan to the observed antibacterial potencies. Antibacterial effects were only detected in F17 and F18 across 20 fractions, which was consistent with the elution of triclosan and triclocarban in the same two fractions. Further, potency mass balance analysis confirmed that triclosan explained the majority (58-113%) of inhibition effects from sludge extracts. This study highlighted triclosan as the predominant antibacterial compound in sewage sludge impacting E. coli despite the co-occurrence of numerous other antibiotics and nonantibiotics.

Acetazolamide Alleviate Cerebral Edema Induced by Ischemic Stroke Through Inhibiting the Expression of AQP4 mRNA.

OBJECTIVE: We want to investigate the effect of aquaporin-4 (AQP4) on cerebral edema induced by ischemic stroke in rats and explore whether inhibiting the expression of AQP4 through acetazolamide (AZA) could attenuate brain edema and protect cerebral function. METHODS: The Sprague Dawley (SD) rats were randomly divided into four groups: sham + saline group, sham + AZA group, AZA intervention group, and nonintervention group. Each group was divided into five subgroups according to the time of cerebral ischemia (6 h, 1 day, 3 days, 5 days, and 7 days). The model of cerebral infarction in rats was adopted by means of the bilateral carotid arteries ligation (2-VO) method. The rats in intervention group were given intraperitoneal injection of AZA (35 mg/kg/day). Hematoxylin-eosin staining was performed for pathological analysis of the infarcted area. The brain water content was calculated to evaluate the degree of brain edema. The messenger RNA (mRNA) and protein expressions of AQP4 in the brain were measured by quantitative real-time polymerase chain reaction and immunohistochemistry, respectively. RESULTS: Significant cerebral pathological damages were found in ischemic stroke rats. The brain water content, protein, and mRNA expression of AQP4 of the intervention and nonintervention groups were markedly higher than those of the sham groups. By contrast, AZA administration reduced the brain water content, whereas improved cerebral dysfunction was induced by ischemic stroke. Moreover, AZA obviously reduced the protein and mRNA expression of AQP4 after ischemic stroke in rats' brains. CONCLUSIONS: The expression of AQP4 was closely related to cerebral edema induced by ischemic stroke. Decreasing the expression of AQP4 mRNA by AZA administration can effectively relieve cerebral edema and decrease cerebral pathological damage.

Sohlh2 inhibits breast cancer cell proliferation by suppressing Wnt/ß-catenin signaling pathway.

Sohlh2 belongs to the superfamily of basic helix-loop-helix (bhlh) transcription factors. Aberrant expression of bhlh transcription factors has been shown to be associated with multiple tumorigenesis. We previously identified that sohlh2 functioned as a tumor suppressor in ovarian cancer. Here, we examined the expression levels of sohlh2 in human breast cancer and its potential role in disease pathogenesis. The results of sohlh2 immunohistochemistry (IHC) and Western blot analysis demonstrated the decreased sohlh2 expression in breast cancer specimens as compared to adjacent noncancerous tissues. Through in vitro MTT, BrdU, colony formation and cell cycle assays and in vivo tumor xenograft studies, we showed that forced expression of sohlh2 led to a significant reduction in proliferation due to G1 arrest in vitro and tumorigenesis in nude mice. Conversely, silencing of sohlh2 enhanced breast cancer cell proliferation. Furthermore, we confirmed that sohlh2 inhibited breast cancer cell proliferation by suppressing the Wnt/ß-catenin signaling pathway. APC was the direct target of sohlh2, and mediated the inhibitory activities of sohlh2 on Wnt/ß-catenin signaling pathway. Thus, our data indicate that sohlh2 likely functions as a tumor suppressor in breast cancer that is mediated by repressing Wnt/ß-catenin signaling pathway via upregulation of APC expression.

Label-free diagnosis of lung cancer with tissue-slice surface-enhanced Raman spectroscopy and statistical analysis.

Despite the rapid development of medical science, the diagnosis of lung cancer is still quite challenging. Due to the ultrahigh detection sensitivity of surface-enhanced Raman spectroscopy (SERS), SERS has a broad application prospect in biomedicine, especially in the field of tumor blood detection. Although Raman spectroscopy can diagnose lung cancer through tissue slices, its weak cross sections are problematic. In this study, silver nanoparticles (AgNPs) were added to the surface of lung tissue slices to enhance the Raman scattering signals of biomolecules. The electromagnetic field distribution of AgNPs prepared was simulated using the COMSOL software. SERS obtained from the slices reflected the difference in biochemical molecules between normal (n = 23) and cancerous (n = 23) lung tissues. Principal component-linear discriminate analysis (PCA-LDA) was utilized to classify lung cancer and healthy lung tissues. The receiver operating characteristic curve gave the sensitivity (95.7%) and specificity (95.7%) of the PCA-LDA method. This study sheds new light on the general applicability of SERS analysis of tissue slices in clinical trials.

Relative chronic sensitivity of neonicotinoid insecticides to Ceriodaphnia dubia and Daphnia magna.

Neonicotinoid insecticides are a group of plant protectants frequently detected in surface waters at low concentrations. Aquatic invertebrates therefore have the potential to be exposed chronically to low concentrations of neonicotinoids. The cladocerans Daphnia magna and Ceriodaphnia dubia are among the most commonly used invertebrate test species in aquatic toxicology. Both species are known to be acutely insensitive to neonicotinoids, and while chronic toxicity has been characterized for D. magna, little research has been conducted with C. dubia. In the present study we conducted 7-d static-renewal life cycle tests for 6 neonicotinoids (acetamiprid, clothianidin, dinotefuran, imidacloprid, thiacloprid, and thiamethoxam) with C. dubia, and a 21-d test with imidacloprid with D. magna. 7-d LC50s for C. dubia ranged from 8.42 mg L-1 for imidacloprid to > 100 mg L-1 for clothianidin; 7-d reproduction EC50s were 2.98 for thiacloprid, to > 67 mg L-1 for dinotefuran. D. magna were less sensitive than C. dubia to imidacloprid, by 4-fold for lethality and 1.5-fold for reproduction; however, acute-to-chronic ratios (ACRs) were similar. ACRs, based on 48-h acute LC50s and 7- or 21-d chronic reproduction EC10s, ranged from 5.4 for acetamiprid to 53.0 for imidacloprid (mean 36.6, CV = 51%). Chronic toxicity values for both species were orders of magnitude greater than concentrations reported in the environment, and thus hazard to these cladocerans is negligible.

Sohlh2 inhibits human ovarian cancer cell invasion and metastasis by transcriptional inactivation of MMP9.

Identifying key mediators of cancer invasion and metastasis is crucial to the development of new and more effective therapies. We previously identified Sohlh2 as an important inhibitor of ovarian cancer cell proliferation. However, the function of Sohlh2 in cell migration and invasion remains unknown. In this paper, we report a novel Sohlh2 to MMP9 signaling pathway in the invasive ovarian cancer. Using immunohistochemistry staining, we revealed Sohlh2 expression was inversely correlated with the invasive human ovarian cancers. In vitro experiments, forced expression of Sohlh2 led to a significant reduction in cancer cell migration and invasion. Conversely, silencing of Sohlh2 enhanced ovarian cancer cell migration and invasion. Experiments using nude mice demonstrated that the ectopic Sohlh2 expression inhibited the HO8910 cell capability of the metastasis to the lungs and livers. Ectopic overexpression of Sohlh2 in the invasive HO8910 cells reduced the MMP9 expression, whereas Sohlh2 knockdown from the non-invasive, SKOV3 cells increased the MMP9 expression. Promoter activation and binding analyses indicated that Sohlh2 repressed the MMP9 expression by directly acting on the MMP9 gene promoter. Inhibition of MMP9 dramatically blocked the Sohlh2 knockdown-enhanced SKOV3 cell invasion, and ectopic expression of MMP9 compensated for the anti-invasive activity of Sohlh2 in HO8910 cells. Overall, these results demonstrate for the first time that Sohlh2 functions as a tumor metastasis suppressor. Modulation of Sohlh2 expression has the potential to be a target for cancer therapy. © 2015 Wiley Periodicals, Inc.

Sphingosine kinase 2 promotes colorectal cancer cell proliferation and invasion by enhancing MYC expression.

Colorectal cancer (CRC) is one of the most commonly diagnosed cancers and causes of cancer death in developed countries. SphK2 is overexpressed in a number of aggressive human carcinomas; however, the expression profile and potential function of SphK2 in CRC are still unknown. In this study, we investigated the SphK2 expression in tumoral tissue and the matched normal mucosae using quantitative real-time PCR (qRT-PCR), Western blot, and immunohistochemistry. We also evaluated the impact of SphK2 knockdown on CRC cell proliferation and metastasis in vitro. SphK2 was significantly upregulated in CRC tissue as compared to the matched normal mucosae, and significant overexpression was found in the LoVo CRC cell line. SphK2 depletion by specific small interfering RNA (siRNA) in the CRC cell line was found to affect cell proliferation and cell migration. Our data suggest that the pathogenesis of CRC maybe mediated by SphK2, and SphK2 could represent a selective target for the molecularly targeted treatments of CRC.

Nitenpyram degradation in finished drinking water.

RATIONALE: Neonicotinoid pesticides and their metabolites have been indicated as contributing factors in the decline of honey bee colonies. A thorough understanding of neonicotinoid toxicity requires knowledge of their metabolites and environmental breakdown products. This work investigated the rapid degradation of the neonicotinoid nitenpyram in finished drinking water. METHODS: Nitenpyram reaction products were characterized using liquid chromatography/quadrupole time-of-flight mass spectrometry (LC/QTOFMS). A software algorithm that compared degraded and control samples was utilized to facilitate efficient data reduction. Fragmentation pathways for six reaction products and nitenpyram were proposed using predictive software and manual product ion analysis. RESULTS: This study showed that nitenpyram degradation in unpreserved finished drinking water was likely the result of oxidation, hydrolysis and reaction with Cl2 . Structures for six nitenpyram reaction products were proposed that suggest the C9/C11 olefin as the key reactive site. CONCLUSIONS: Similarities between the identified nitenpyram reaction products and imidacloprid metabolites highlight the importance of this study, as the toxicity of neonicotinoids to pollinators has been linked to their metabolites. Based on the proposed reaction mechanisms, the identified nitenpyram reaction products in finished drinking water could also be present in the environment and water treatment facilities. As such, identifying these degradation products will aid in evaluating the environmental impact of neonicotinoid pesticides. Copyright © 2016 John Wiley & Sons, Ltd.

Liquid chromatography/tandem mass spectrometry analysis of neonicotinoids in environmental water.

RATIONALE: Neonicotinoids (NNIs) are the fastest expanding group of pesticides in the world over the last two decades; however, they may be a significant contributing factor to bee mortality. The widespread use of NNIs makes it critical to monitor their residuals in the environment. Published methods for NNI analysis are mainly focused on agricultural and food products, and many of them only measured a portion of the commercially available NNIs. METHODS: Utilizing a biphenyl stationary-phase column, a sensitive liquid chromatography/tandem mass spectrometry (LC/MS/MS) method was developed to determine eight NNIs, including acetamiprid, clothianidin, dinotefuran, flonicamid, imidacloprid, nitempyram, thiacloprid and thiamethoxam in environmental water. Two multiple reaction monitoring (MRM) transitions were monitored for each compound to ascertain true positive identification. Isotope-labelled NNIs, d3-acetamiprid, d3-clothianidin, d4-imidacloprid and d3-thiamethoxam, were used to compensate for extraction efficiency, matrix effects and instrument variability while monitoring real-time method performance. Target compounds in aqueous samples were analyzed by direct aqueous injection (DAI) or after solid-phase extraction (SPE). RESULTS: The method detection limits (MDLs) of NNIs in drinking water, surface water and groundwater were in the ranges of 50 to 190 and 2 to 7 ng/L for DAI and SPE procedures, respectively, and target compound recoveries ranged from 78 to 110%. The stability of target compounds in water samples and SPE extracts was also investigated for the first time to ensure accurate results. No obvious degradation was observed for target compounds within four weeks in either water samples or SPE extracts. CONCLUSIONS: The method developed for neonicotinoid pesticide analysis is very sensitive and efficient. It provides good flexibility to meet various environmental monitoring needs and is employed for an extensive study to determine the distribution of NNIs in Ontario's water.

Interleukin-28B Plays a Therapeutic Role on Mouse U14 Cervical Cancer Cells by Down-Regulating CD4+CD25+FoxP3+Regulatory T Cells In Vivo.

AIM: To investigate the immunotherapeutic effectiveness of adenoviral vector expressing mouse interleukin (IL)-28B (Ad-mIL-28B) against cervical cancer and its mechanism. METHOD: U14 cervical cancer cell-bearing mice were treated with Ad-mIL-28B. Meanwhile, whole cell vaccine was prepared by repeated freezing and thawing U14 cells. Then CD4⁺CD25⁺FoxP3⁺regulatory T (Treg) cells were evaluated by flow cytometry. Tumor volume and metastasis in BALB/c and C57BL/6j mice were detected. RESULTS: Ad-mIL-28B treatment significantly decreased the number of CD4⁺CD25⁺FoxP3⁺Treg cells. Subsequently, there was a significant decrease in the size of tumor tissue and the numbers of heteromorphic tumor cells. The tumor metastasis in the lung and liver of the Ad-mIL-28B group also decreased. However, there was no therapeutic effect observed for whole cell vaccine on U14 tumor-bearing mice. CONCLUSION: Interleukin-28B can inhibit the growth and metastasis of cervical cancer in U14 tumor-bearing mice by down-regulating Treg cells.

Sohlh2 inhibits ovarian cancer cell proliferation by upregulation of p21 and downregulation of cyclin D1.

Spermatogenesis and oogenesis basic helix-loop-helix (bHLH) transcription factor 2 (Sohlh2) functions as a bhlh transcription factor to regulate mouse germ cell differentiation. Our previous data showed that Sohlh2 was highly expressed in human normal tissues, but low level of Sohlh2 was observed in many cancer cell lines, suggesting a possible role of Sohlh2 in tumorigenesis. In this study, we examined this possibility by using immunohistochemistry, MTT, 5-bromo-2-deoxyuridine, clonogenic assay and tumor xenograft techniques. Our results showed that the expression of Sohlh2 was decreased in epithelial ovarian carcinoma (EOC) tissues compared with benign ovarian tumors and ovarian tumors with low malignant potential. Forced expression of Sohlh2 led to a significant reduction in cancer cell proliferation in vitro and tumorigenesis in nude mice. Conversely, silencing of Sohlh2 enhanced ovarian cancer cell proliferation. Furthermore, Sohlh2 had opposite effects on its two direct targets p21 and cyclin D1: overexpression of Sohlh2 upregulated p21 but downregulated cyclin D1 expression. p21 knockdown could reverse the effects of Sohlh2 overexpression on inhibiting cell proliferation, and cyclin D1 knockdown could reverse the effects of Sohlh2 ablation on promoting cell proliferation. Thus, our data indicate that Sohlh2 likely functions as a tumor suppressor in EOCs, which is achieved by inducing p21 expression but repressing cyclin D1 expression.

Tumor-stroma ratio is an independent predictor for survival in early cervical carcinoma.

OBJECTIVE: Tumor-stroma ratio (TSR) has recently been identified as an independent prognostic parameter for several solid tumors. The aim of the present study was to evaluate the prognostic role of TSR in early cervical cancer. METHODS: A cohort of 184 patients who had surgery for early stage cervical cancer (FIGO [International Federation of Gynecology and Obstetrics] stages IA2-IIA) were enrolled in this study. TSR was estimated on hematoxylin-eosin-stained tissue sections from the most invasive part of the primary tumor. Patients with less than 50% stroma were classified as stroma-poor and patients with ≥ 50% stroma were classified as stroma-rich. The relationship between TSR and survival time was statistically analyzed. RESULTS: The disease-free survival and overall survival rates were 88.44% and 92.52%, respectively, in the stroma-poor group, and 62.16% and 70.27%, respectively, in the stroma-rich group. Both the disease-free and overall survival rates in the stroma-poor group were significantly better than those in the stroma-rich group (p=0.001). In a multivariate analysis, TSR was further confirmed as a significant prognostic factor for disease-free survival (hazard ratio 3.125; p=0.005) and overall survival (hazard ratio 3.464; p=0.003), independent of tumor size, FIGO stage and lymph node metastasis. CONCLUSION: Our study identified that TSR was an independent prognostic factor of early cervical cancer. Patients with stroma-rich tumors had worse prognosis and higher risk of relapse compared with those with stroma-poor tumors. Considering its simplicity and availability for conventional clinical pathology, TSR may serve as a new prognostic histological characteristic in early cervical cancer.

The response of amphibian larvae to exposure to a glyphosate-based herbicide (Roundup WeatherMax) and nutrient enrichment in an ecosystem experiment.

Herbicides and fertilizers are widely used throughout the world and pose a threat to aquatic ecosystems. Using a replicated, whole ecosystem experiment in which 24 small wetlands were split in half with an impermeable barrier we tested whether exposure to a glyphosate-based herbicide, Roundup WeatherMax™, alone or in combination with nutrient enrichment has an effect on the survival, growth or development of amphibians. The herbicide was applied at one of two concentrations (low=210 µg a.e./L, high=2880 µg a.e./L) alone and in combination with nutrient enrichment to one side of wetlands and the other was left as an untreated control. Each treatment was replicated with six wetlands, and the experiment was repeated over two years. In the high glyphosate and nutrient enrichment treatment the survival of wood frog (Lithobates sylvaticus) larvae was lower in enclosures placed in situ on the treated sides than the control sides of wetlands. However, these results were not replicated in the second year of study and they were not observed in free swimming wood frog larvae in the wetlands. In all treatments, wood frog larvae on the treated sides of wetlands were slightly larger (<10%) than those on the control side, but no effect on development was observed. The most dramatic finding was that the abundance of green frog larvae (Lithobates clamitans) was higher on the treated sides than the control sides of wetlands in the herbicide and nutrient treatments during the second year of the study. The results observed in this field study indicate that caution is necessary when extrapolating results from artificial systems to predict effects in natural systems. In this experiment, the lack of toxicity to amphibian larvae was probably due to the fact the pH of the wetlands was relatively low and the presence of sediments and organic surfaces which would have mitigated the exposure duration.

The role of neuropeptide Y and aquaporin 4 in the pathogenesis of intestinal dysfunction caused by traumatic brain injury.

BACKGROUND: Although the exact incidence is unknown, traumatic brain injury (TBI) can lead to intestinal dysfunction. It has important influence on the early nutrition and prognosis of TBI patients. Experiments were designed to study the roles of neuropeptide Y (NPY) and aquaporin 4 (AQP4) in the pathogenesis of intestinal dysfunction caused by TBI and to find some new solutions for the treatment of intestinal dysfunction after TBI. METHODS: Forty adult male Wistar rats were randomly divided into control, mild trauma, moderate trauma, and severe trauma groups. TBI was induced by Feeney's impact method. Control animals were sham operated but not subjected to the impact test. All rats were killed 24 h after surgery. Blood samples were obtained from the abdominal aorta for enzyme-linked immunosorbent assay measurement of NPY concentrations. Jejunum segments 15 cm distal to the Treitz ligament were taken for analysis of NPY and AQP4 expression by polymerase chain reaction, Western blot, and immunohistochemistry. Pathologic changes in intestinal cell structure and ultrastructure were studied by light microscopy and transmission electron microscopy. RESULTS: The specimens from different groups showed different degrees of structural changes, ranging from swelling and degeneration of villous epithelial cells to extensive denudation and collapse of the villi. The more severe the trauma, the more serious the degree of intestinal mucosal injury. Intestinal smooth muscle also showed varying degrees of edema and structural disorder. Electron microscopy showed that intestinal mitochondria had varying degrees of swelling and the structure of mitochondrial crista was disordered and even fractured. Plasma concentrations of NPY and jejunal gene and protein expressions of NPY and AQP4 increased significantly following TBI (P < 0.05), with greater increases at higher levels of injury. Moreover, there were positive correlations between NPY and AQP4 (P < 0.05). CONCLUSIONS: Increasing grades of TBI caused increasing degrees of intestinal ischemia and edema, and thus caused increasingly severe intestinal dysfunction. AQP4 and NPY may be involved in the pathogenesis of intestinal dysfunction after TBI. Increased NPY levels may be responsible for intestinal ischemia and hypoxia, and AQP4 may play an important role in intestinal edema. Increased NPY levels may be one of the main causes for the increase in AQP4 after TBI.

A novel oncolytic virus-based biomarker participates in prognosis and tumor immune infiltration of glioma.

Background: Glioma is the most common central nervous malignancy. Due to its poor survival outcomes, it is essential to identify novel individualized therapy. Oncolytic virus (OV) treatment is a key therapy regulating tumor microenvironment in malignant glioma. Herein, we aim to identify the key genes after OV infection and its role in glioma. Methods: Performing an RNA-seq analysis, the differentially expressed genes (DEGs) between EV-A71-infection and mock group were screened with GFold values. DAVID online analysis was performed to identify the functional classification. Overall survival (OS) or disease-free survival (DFS) was evaluated to analyze the relation between PTBP1 expression levels and prognosis of glioma patients. Additionally, the ssGSEA and TIMER algorithms were applied for evaluating immune cell infiltration in glioma. Results: Following EV-A71 infection in glioma cells, PTBP1, one of the downregulated DEGs, was found to be associated with multiple categories of GO and KEGG enrichment analysis. We observed elevated expression levels of PTBP1 across various tumor grades of glioma in comparison to normal brain samples. High PTBP1 expression had a notable impact on the OS of patients with low-grade glioma (LGG). Furthermore, we observed an obvious association between PTBP1 levels and immune cell infiltration in LGG. Notably, PTBP1 was regarded as an essential prognostic biomarker in immune cells of LGG. Conclusion: Our research uncovered a critical role of PTBP1 in outcomes and immune cell infiltration of glioma patients, particularly in those with LGG.

A Novel Prognostic Marker and Therapeutic Target Associated with Glioma Progression in a Tumor Immune Microenvironment.

Background: Immune microenvironment serves a vital role in glioma progression, and a large number of studies have found that tumor progression can be reduced to some extent by modulating the immune process in tumors. Materials and Methods: ImmuneScore of each sample in CGGA datasets were calculated with Estimate R package, and samples were grouped by median ImmuneScore values for differential analysis to obtain immune microenvironment differential genes. We further conducted survival analysis, ROC curve analysis, independent prognostic analysis, and clinical correlation analysis on glioma sample genes in CGGA to obtain glioma prognostic genes, and then identified their intersection with immune microenvironment DEGs by Venn tool. The GEPIA and UALCAN databases were used to verify the differential expression of intersecting genes in the glioma and normal brain and to identify our target gene. After validation of their prognostic value, we constructed a nomogram to calculate the risk score and to estimate the accuracy of prognostic model. We mined co-expression genes, enriched functions and pathways, and correlations to immune cell infiltration of unigene with an online database. Finally, we verified the differential expression of FCGBP in glioma by immunohistochemical staining. Results: We finally selected Fc fragment of IgG-binding protein (FCGBP) as our study gene. The prognostic values of FCGBP were validated by a series of analyses. Immunohistochemical staining showed that FCGBP expression increased in gliomas and was up-regulated with the progression of glioma grade. Conclusion: As a key unigene in glioma progression, FCGBP contributes to the regulation of immune microenvironment and has the potential to be a prognostic biomarker and immune targets.