FAM84B promotes the proliferation of glioma cells through the cell cycle pathways.

BACKGROUND: This study aimed to investigate FAM84B expression in glioma tissues and explore the role of FAM84B in promoting the proliferation of glioma cells and the mechanism of regulating the cell cycle pathways. METHODS: The TCGA database was adopted to analyze FAM84B expression in glioma tissues. The FAM84B expression was detected by qRT-PCR in patients with glioma, especially that in glioma cells, U251, LN-229, U98, and U87. Two glioma cell lines U87 and T98 were selected for siRNA transfection, which were divided into si-NC si-FAM84B-1 and si-FAM84B-2 groups. The effect of FAM84B on the proliferation of glioma cells was detected with the MTT experiment and that on the glioma cell cycle was detected with the flow cytometry. The signaling pathways potentially regulated by FAM84B in glioma were analyzed through the bioinformatics analysis. The expression of proteins, Cyclin D1, CDK4, Cdk6, and p21, in the cell cycle-related pathways in cells of each group was detected by the Western blot. RESULTS: TCGA database results showed a significantly higher FAM84B expression in glioma tissues than that in paracancerous tissues. According to the detection of qRT-PCR, FAM84B expressed the highest in the glioma cell line U87 (P < 0.05). Compared with the serum of healthy controls, FAM84B mRNA expression significantly increased in patients with gliomas. And compared with the si-NC group, the proliferation ability of U87 and T98 cells decreased and the cell cycle was blocked in the G0/G1 phase in both si-FAM84B transfection groups (P < 0.05). According to the bioinformatics analysis, FAM84B regulated the cell cycle pathways in glioma. FAM84B siRNA inhibited the expression of key proteins, Cyclin D1, CDK2, CDK4, and Cdk6, of the cell cycle pathways in glioma cells and promoted the expression of P53 and P21 proteins. CONCLUSIONS: In conclusion, FAM84B may inhibit the proliferation of glioma cells by regulating the cell cycle pathways.

Validation of a HRGC-ECNI/LRMS method to monitor short-chain chlorinated paraffins in human plasma.

Short-chain chlorinated paraffins (SCCPs) are produced in high volume and have the high potential to pose a threat to human health. However, little information is available for SCCP contamination in human blood/plasma/serum, mainly due to the difficulty of sample preparation and quantitative analysis. A method using high resolution gas chromatography coupled with electron capture negative ionization low resolution mass spectrometry (HRGC-ECNI/LRMS) was developed and validated to measure SCCPs in human plasma. The pretreatment process included protein denaturation and lipid elimination, liquid-liquid extraction with a mixture of n-hexane/dichloromethane (1:1, V/V), and cleanup on a multi-layer silica column. The blank controls, including procedural blank, vacuum blood collection tube blank, and instrumental blank, were the most pivotal points for the reliable analysis of SCCPs. The average value of procedural blanks was 9.0ng/g; and the method detection limit (MDL), calculated as the sum of the average procedural blank value and 3 times of the standard deviation of the procedural blanks, was 12.6ng/g plasma. The validated method was applied to measure the concentrations of the total SCCPs (∑SCCPs) in 50 plasma samples from a general population. The measured plasma concentrations of ∑SCCPs ranged from

Release and Gas-Particle Partitioning Behaviors of Short-Chain Chlorinated Paraffins (SCCPs) During the Thermal Treatment of Polyvinyl Chloride Flooring.

Chlorinated paraffin (CP) mixture is a common additive in polyvinyl chloride (PVC) products as a plasticizer and flame retardant. During the PVC plastic life cycle, intentional or incidental thermal processes inevitably cause an abrupt release of short-chain CPs (SCCPs). In this study, the thermal processing of PVC plastics was simulated by heating PVC flooring at 100-200 °C in a chamber. The 1 h thermal treatment caused the release of 1.9-10.7% of the embedded SCCPs. A developed emission model indicated that SCCP release was mainly controlled by material-gas partitioning at 100 °C. However, release control tended to be subjected to material-phase diffusion above 150 °C, especially for SCCP congeners with shorter carbon-chain lengths. A cascade impactor (NanoMoudi) was used to collect particles of different sizes and gas-phase SCCPs. The elevated temperature resulted in a higher partition of SCCPs from the gas-phase to particle-phase. SCCPs were not strongly inclined to form aerosol particles by nucleation, and less present in the Aitken mode particles. Junge-Pankow adsorption model well fitted the partitioning of SCCPs between the gas-phase and accumulation mode particles. Inhalation exposure estimation indicated that PVC processing and recycling workers could face a considerably high risk for exposure to SCCPs.

Dispersion of Short- and Medium-Chain Chlorinated Paraffins (CPs) from a CP Production Plant to the Surrounding Surface Soils and Coniferous Leaves.

Chlorinated paraffin (CP) production is one important emission source for short- and medium-chain CPs (SCCPs and MCCPs) in the environment. In this study, 48 CP congener groups were measured in the surface soils and coniferous leaves collected from the inner and surrounding environment of a CP production plant that has been in operation for more than 30 years to investigate the dispersion and deposition behavior of SCCPs and MCCPs. The average concentrations of the sum of SCCPs and MCCPs in the in-plant coniferous leaves and surface soils were 4548.7 ng g-1 dry weight (dw) and 3481.8 ng g-1 dw, which were 2-fold and 10-fold higher than those in the surrounding environment, respectively. The Gaussian air pollution model explained the spatial distribution of CPs in the coniferous leaves, whereas the dispersion of CPs to the surrounding surface soils fits the Boltzmann equation well. Significant fractionation effect was observed for the atmospheric dispersion of CPs from the production plant. CP congener groups with higher octanol-air partitioning coefficients (KOA) were more predominant in the in-plant environment, whereas the ones with lower KOA values had the elevated proportion in the surrounding environment. A radius of approximately 4 km from the CP production plant was influenced by the atmospheric dispersion and deposition of CPs.

Identification of Family with Sequence Similarity 110 Member C (FAM110C) as a Candidate Diagnostic and Prognostic Biomarker for Glioma.

Background: Gliomas are the most frequent and dangerous primary cerebral tumors. Therefore, there is a need to develop molecular targets for the diagnosis and treatment for glioma. Methods: In September 2020, we retrieved the expression matrix of glioblastoma (GBM) sufferers and pertinent clinical data from the TCGA (The Cancer Genome Atlas) database. Prognostic differences between various families with sequence similarity 110 member C (FAM110C) expression groups were assessed by Kaplan-Meier with log-rank test. The R platform get used to assess the accuracy of FAM110C delivery in predicting the prognosis of PDAC using a time-dependent receptor operating characteristic (ROC) curve. The delivery level of FAM110C was determined by qRT-PCR and western blot. Gene set enrichment investigated possible mechanisms between different FAM110C expression groups in GBM (GSEA). The impact of FAM110C on glioma cell movement was discovered using migration test. The drug's gene-targeting impact was validated by the CCK8 test. Results: A total of 173 GBM samples were obtained from the TCGA database, with 148 including information on IDH1 mutations and 151 containing information on overall survival. The mRNA expression level of FAM110C was greater in wild-type GBM, according to qRT-PCR data. The connection between FAM110C expression and Hallmark, GO, and KEGG pathway gene sets was investigated using GSEA software. We used migration test to assess the impact of FAM110C on glioma motility in order to confirm the findings of the GSEA analysis. Conclusion: FAM110C might get used as a possible diagnostic and prognostic biomarker for wild-type GBM, and its inhibition could be used to prevention and treatment wild-type GBM.

ATG14 plays a critical role in hepatic lipid droplet homeostasis.

BACKGROUND & AIMS: Autophagy-related 14 (ATG14) is a key regulator of autophagy. ATG14 is also localized to lipid droplet; however, the function of ATG14 on lipid droplet remains unclear. In this study, we aimed to elucidate the role of ATG14 in lipid droplet homeostasis. METHODS: ATG14 loss-of-function and gain-of-function in lipid droplet metabolism were analyzed by fluorescence imaging in ATG14 knockdown or overexpression hepatocytes. Specific domains involved in the ATG14 targeting to lipid droplets were analyzed by deletion or site-specific mutagenesis. ATG14-interacting proteins were analyzed by co-immunoprecipitation. The effect of ATG14 on lipolysis was analyzed in human hepatocytes and mouse livers that were deficient in ATG14, comparative gene identification-58 (CGI-58), or both. RESULTS: Our data show that ATG14 is enriched on lipid droplets in hepatocytes. Mutagenesis analysis reveals that the Barkor/ATG14 autophagosome targeting sequence (BATS) domain of ATG14 is responsible for the ATG14 localization to lipid droplets. Co-immunoprecipitation analysis illustrates that ATG14 interacts with adipose triglyceride lipase (ATGL) and CGI-58. Moreover, ATG14 also enhances the interaction between ATGL and CGI-58. In vitro lipolysis analysis demonstrates that ATG14 deficiency remarkably decreases triglyceride hydrolysis. CONCLUSIONS: Our data suggest that ATG14 can directly enhance lipid droplet breakdown through interactions with ATGL and CGI-58.

The anti-tumor effect of proteasome inhibitor MG132 for human adenoid cystic carcinoma: correlate with the emerging role of Nrf2/Keap1 signaling pathway.

BACKGROUND: Adenoid cystic carcinoma (ACC) is one of the most common malignant salivary gland tumors. Moreover, the unique biological characteristics and complex structures of ACC contribute to its poor survival rates. Recently, proteasome inhibitors have been shown to elicit satisfactory therapeutic effects in the treatment of certain solid tumors, but few studies have been implemented to investigate the effects of proteasome inhibitor therapy for ACC. METHODS: In this present study, cell counting kit-8 assay and flow cytometry assay were performed to examine the effects of proteasome inhibitor (MG132) on cell viability and apoptosis. We applied western blot and immunofluorescence staining to explore the expression of the Nrf2/Keap1 signaling pathway and P62, additionally Nrf2 inhibitor (ML385) was utilized to evaluate the role of Nrf2/Keap1 signaling pathway in MG132-induced cell apoptosis. RESULTS: Our data indicated that MG132 significantly suppressed the growth of ACC-83 cells(MG132 10µM P = 0.0046; 40µM P = 0.0033; 70µM P = 0.0007 versus control) and induced apoptosis (MG132 10µM P = 0.0458; 40µM P = 0.0018; 70µM P = 0.0087 versus control). The application of MG132 induced the up-regulation of Nrf2/Keap1 signaling pathway. Furthermore, inhibition of Nrf2 attenuated the therapeutic effects of MG132 for ACC (both ML385 + MG132 10µM P = 0.0013; 40µM P = 0.0057; 70µM P = 0.0003 versus MG132). P < 0.05 was considered statistically significant. CONCLUSIONS: Our results revealed that proteasome inhibitors MG132 could inhibit the cell viability and induce the apoptosis of ACC through Nrf2/Keap1 signaling pathway.

Genetically programmed alternative splicing of NEMO mediates an autoinflammatory disease phenotype.

Host defense and inflammation are regulated by the NF-κB essential modulator (NEMO), a scaffolding protein with a broad immune cell and tissue expression profile. Hypomorphic mutations in inhibitor of NF-κB kinase regulatory subunit gamma (IKBKG) encoding NEMO typically present with immunodeficiency. Here, we characterized a pediatric autoinflammatory syndrome in 3 unrelated male patients with distinct X-linked IKBKG germline mutations that led to overexpression of a NEMO protein isoform lacking the domain encoded by exon 5 (NEMO-Δex5). This isoform failed to associate with TANK binding kinase 1 (TBK1), and dermal fibroblasts from affected patients activated NF-κB in response to TNF but not TLR3 or RIG-I-like receptor (RLR) stimulation when isoform levels were high. By contrast, T cells, monocytes, and macrophages that expressed NEMO-Δex5 exhibited increased NF-κB activation and IFN production, and blood cells from these patients expressed a strong IFN and NF-κB transcriptional signature. Immune cells and TNF-stimulated dermal fibroblasts upregulated the inducible IKK protein (IKKi) that was stabilized by NEMO-Δex5, promoting type I IFN induction and antiviral responses. These data revealed how IKBKG mutations that lead to alternative splicing of skipping exon 5 cause a clinical phenotype we have named NEMO deleted exon 5 autoinflammatory syndrome (NDAS), distinct from the immune deficiency syndrome resulting from loss-of-function IKBKG mutations.

Artesunate Combined With Metformin Ameliorate on Diabetes-Induced Xerostomia by Mitigating Superior Salivatory Nucleus and Salivary Glands Injury in Type 2 Diabetic Rats via the PI3K/AKT Pathway.

Polydipsia and xerostomia are the most common complications that seriously affect oral health in patients with diabetes. However, to date, there is no effective treatment for diabetic xerostomia. Recent studies have reported that artesunate (ART) and metformin (Met) improve salivary gland (SG) hypofunction in murine Sjögren's syndrome. Therefore, aim of this study was to investigate the effect and underlying mechanism of artesunate (ART) alone and in combination with metformin (Met) on hyposalivation in type 2 diabetes mellitus (T2DM) rats. T2DM rats were induced using a high-fat diet and streptozotocin. SPF male Sprague-Dawley rats were divided into the following five groups: normal control group, untreated diabetic group, ART-treated diabetic group (50 mg/kg), Met-treated diabetic group (150 mg/kg), and ART/Met co-treated diabetic group (50 mg/kg ART and 150 mg/kg Met). ART and Met were intragastrically administered daily for 4 weeks. The general conditions, diabetes parameters and serum lipids were evaluated after drug treatment. Furthermore, we observed changes in the central superior salivatory nucleus (SSN) and SG, and changes in the AQP5 expression, parasympathetic innervation (AChE and BDNF expression), and PI3K/AKT pathway- (p-AKT, and p-PI3K), apoptosis- (Bax, Bcl-2, and Caspase3), and autophagy- (LC3 and P62) related markers expression in T2DM rats after treatment. Our results showed that ART or Met alone and ART/Met combination attenuated a range of diabetic symptoms, including weight loss, urine volume increase, water consumption increase, hyperglycemia, insulin resistance, glucose intolerance and dyslipidemia. More importantly, we found that these three treatments, especially ART/Met combination, mitigated hyposalivation in the T2DM rats via improving the central SSN and SGs damage in hyperglycemia. Our data also indicated that ART/Met attenuated SG damage though regulating the PI3K/Akt pathway to inhibit apoptosis and autophagy of SGs in the T2DM rats. Moreover, ART/Met preserved parasympathetic innervation (AChE and BDNF expression) in SGs to alleviate diabetes-induced hyposalivation likely through rescuing central SSN damage. Taken together, these findings might provide a novel rationale and treatment strategy for future treatment of diabetes-induced xerostomia in the clinic.

Circ-EZH2 knockdown reverses DDAH1 and CBX3-mediated cell growth and invasion in glioma through miR-1265 sponge activity.

Accumulating evidence has indicated the important roles of circular RNAs (circRNAs) in different tumors. However, their detailed regulatory mechanisms in glioma are not fully understood. In this study, the functional role of a novel circRNA, circ-EZH2, was investigated by cell counting kit-8 (CCK-8), colony formation, flow cytometry, and transwell experiments. The regulatory mechanism of circ-EZH2 was explored by bioinformatics analysis, quantitative real-time PCR (qRT-PCR), Western blot and dual-luciferase reporter assay. We identified that circ-EZH2 was overexpressed in glioma tissues and cell lines. Further studies revealed that ectopic expression of circ-EZH2 significantly promoted cell growth, migration and invasion but inhibited cell apoptosis. By contrast, silencing of circ-EZH2 induced the opposite effects. Additionally, we found circ-EZH2 served as a miRNA sponge for miR-1265 to release its suppression on DDAH1 and CBX3. Rescue assays further revealed that the oncogenic function of circ-EZH2 was partly dependent on its modulation of DDAH1 and CBX3. Our study unraveled a novel molecular pathway in glioma and may provide a new perspective for the treatment of glioma.

Paeoniae radix alba polysaccharides obtained via optimized extraction treat experimental autoimmune hepatitis effectively.

The extraction process of Paeoniae radix alba polysaccharides (PRAP) was optimized as the liquid-solid ratio of 10.65 mL/g, the extraction time of 2.10 h, and the 2 extraction repetitions through a response surface methodology. The chemical profiles of the obtained PRAP were characterized by measuring the contents of total carbohydrates, total phenolics, uronic acid and protein, and by analyzing the FT-IR spectrum and monosaccharide composition. To determine the therapeutic effects of PRAP on experimental autoimmune hepatitis (EAH), we established an EAH mice model. After treated with PRAP, liver and spleen injuries were reduced, and hepatocyte regeneration and liver function were improved. Further study of the mechanism by which PRAP treats EAH showed that PRAP significantly inhibited oxidative stress in the livers of EAH model mice. More importantly, PRAP inhibited immune inflammatory reactions in EAH model mice, including the hepatic infiltration of inflammatory CD4+ and CD8+ T cells, as well as overexpression of inflammatory cytokines IL-2, IL-6 and IL-10, via inhibition of the NF-κB signaling pathway.

Integration of metabolomics and transcriptomics reveals short-chain chlorinated paraffin-induced hepatotoxicity in male Sprague-Dawley rat.

BACKGROUND: Short-chain chlorinated paraffins (SCCPs) used in various industrial applications have been listed as new POPs. Previous studies based on high-dose exposures indicate their hepatotoxicity. However, their mechanisms of toxicity or adverse outcome pathways and health risks remain largely unknown. OBJECTIVES: This study aimed to evaluate metabolic consequences of chronic dietary exposure to SCCPs at low doses and reveal the molecular mechanisms underlying hepatotoxicity of SCCPs. METHODS: A combination of transcriptomics and metabolomics, together with general pathophysiological tests were performed to assess the hepatic response of male rats exposed to SCCPs. RESULTS: Our results highlight two major modes of action: Inhibition of energy metabolism and activation of the peroxisome proliferator-activated receptor α (PPARα). Exposure to SCCPs suppressed oxidative phosphorylation, glycolysis, gluconeogenesis and turnover of ATP-ADP-AMP and thus results in deficiencies of amino acids and nucleotides in liver of the rat. Exposure to SCCPs affected expression levels of 13 genes downstream of PPARα that encode proteins associated with metabolism of fatty acids. As a result, peroxisomal and mitochondrial fatty acid ß-oxidation, microsomal fatty acid ω-oxidation, and lipogenesis were accelerated. CONCLUSIONS: Results of this work strongly support the conclusion that low-dose exposure to SCCPs can result in adverse outcomes in the rat model. Significant SCCP-induced inhibition of energy metabolism occurs at environmentally relevant dosages, which suggests that SCCPs exhibit metabolic toxicity. Interactions of SCCPs with PPARα signaling pathway can explain the disruption of lipids and amino acids metabolism.

Concentrations of short- and medium-chain chlorinated paraffins in indoor dusts from malls in China: Implications for human exposure.

Levels and distribution of short- and medium-chain chlorinated paraffins (SCCPs and MCCPs) were measured in indoor dusts from malls in China. The concentrations of SCCPs and MCCPs in dustfalls from a building material mall ranged from 6.0 to 361.4 µg g-1 and from 5.0 to 285.9 µg g-1, respectively. Much heavier contamination was found in central air conditioner filter (CACF) dusts from a newly opened shopping mall, with SCCP concentrations of 114.7-707.0 µg g-1 and MCCP concentrations of 89.0-1082.9 µg g-1. The C13- and C14-CPs were the dominant congeners, while the Cl7 and Cl8 groups were the major chlorine congeners in both kinds of dust samples. Significant correlation relationships (p ≤ 0.05) were found between ∑SCCPs and ∑MCCPs in CACF dusts and dustfalls. Varied exposure pathways including dust ingestion and dermal permeation have been evaluated. The average daily exposure doses of SCCPs and MCCPs for the adult in CACF dusts and dustfalls were estimated to be 0.394 and 0.150 µg kg-1 day-1, respectively. The toddler had higher exposure risks with 5.918 and 2.658 µg kg-1 day-1 in the shopping and building material malls, respectively. Dermal permeation was the predominated exposure pathway for the adult, while dust ingestion was suggested to be more important for the toddler due to hand-to-mouth contact.

High performance solid-phase extraction cleanup method coupled with gas chromatography-triple quadrupole mass spectrometry for analysis of polychlorinated naphthalenes and dioxin-like polychlorinated biphenyls in complex samples.

A solid-phase extraction (SPE) cleanup method was developed to purify the sample extracts for the analysis of polychlorinated naphthalenes (PCNs) and dioxin-like polychlorinated biphenyls (dl-PCBs). Monodisperse magnesium oxide (MgO) microspheres and basic alumina were used as SPE adsorbents. Important parameters of the SPE procedure were optimized, including the amount of basic alumina and the type and volume of the washing and elution solvents. The optimized SPE cleanup method exhibited excellent purification performance for the removal of organochlorinated compounds, lipid compounds, sulfur, and pigments. Additionally, it was found that the retention activities of congeners differed with the number and position of the chlorine substituents in PCNs. In this study, an analytical method based on a combination of accelerated solvent extraction (ASE) coupled with SPE cleanup and gas chromatography-triple quadrupole mass spectrometry (GC-MS/MS) is proposed for the analysis of PCNs and dl-PCBs in complex samples (sediment, pine needle, and scallop samples). The analytical method demonstrates good linearity, acceptable recovery (63-148%) and precision (relative standard deviations less than 26%). The limits of detection (LODs) of PCN and dl-PCB congeners were in the range of 0.6-19.1pgg(-1) and 0.4-8.6pgg(-1), respectively. The PCNs and dl-PCBs levels in these samples ranged from 0.16 to 3.07ngg(-1) dry weight (dw) and from undetectable to 0.07ngg(-1) dw, respectively.