Plasma proteomic analysis reveals altered protein abundances in HIV-infected patients with or without non-Hodgkin lymphoma.

The identification of circulating proteins associated with acquired immunodeficiency syndrome-related non-Hodgkin lymphoma (AIDS-NHL) may help in the development of promising biomarkers for screening, diagnosis, treatment, and prognosis. Here, we used quantitative liquid chromatography-tandem mass spectrometry (LC-MS/MS) to identify differentially expressed proteins (DEPs) in plasma collected from patients with AIDS-NHL and human immunodeficiency virus (HIV)-infected patients without NHL (HIV+ ). Proteins with a log2 (fold change) in abundance >0.26 and p < 0.05 were considered differentially abundant. In total, 84 DEPs were identified, among which 20 were further validated as potential biomarkers, with immunoglobulin and complement components being the most common proteins. Some of the proteins were further verified in a retrospective analysis of the medical records of patients in a larger cohort. These markedly altered proteins were found to mediate pathophysiological pathways that likely contribute to AIDS-NHL pathogenesis, such as the humoral immune response, complement activation, and complement and coagulation cascades. Our findings provide a new molecular understanding of AIDS-NHL pathogenesis and provide new evidence supporting the identification of these proteins as possible biomarkers in AIDS-NHL.

Effects of the Notch Signaling Pathway on Secondary Brain Changes Caused by Spinal Cord Injury in Mice.

Spinal cord injury (SCI) can cause secondary brain changes, leading to hypomyelination in the dorsolateral prefrontal cortex (dlPFC). Some studies have shown that notch signaling pathway activation can regulate oligodendrocyte maturation and myelination. The aim of this study was to investigate whether inhibition of the Notch signaling pathway can alleviate hypomyelination in the dlPFC caused by SCI. Moreover, we further investigated whether the changes in myelination in the dlPFC are associated with neuropathic pain following SCI. We established a mouse model of SCI and observed the changes in mechanical and thermal hyperalgesia. Western blotting and immunofluorescence were used to analyze the changes in myelination in the dlPFC. The results indicated the existence of a relationship between activation of the Notch signaling pathway and hypomyelination in the dlPFC and confirmed the existence of a relationship between hypomyelination in the dlPFC and decreases in mechanical and thermal hyperalgesia thresholds. In conclusion, these results suggested that the Notch signaling pathway is activated after SCI, leading to hypomyelination in the dlPFC, and that DAPT can inhibit the Notch signaling pathway and improve mechanical and thermal hyperalgesia thresholds. Our findings provide a new target for the treatment of neuropathic pain caused by SCI.

Demonstration of a terahertz multi-spectral 3×3 Mueller matrix polarimetry system for 2D and 3D imaging.

Mueller matrix polarimetry (MMP) has been demonstrated and recognized as an effective approach to attaining imaging enhancement as well as revealing polarization properties of an imaged sample. Generally, a minimum of 16 combinations of intensity-only measurements involving both linear and circular polarizations are required to completely and accurately determine the 4 × 4 Mueller matrix (MM) and comprehensively describe the polarization properties of the sample. However, broadband circular polarizations (CP) are rather difficult to obtain for design and fabrication limitations in the terahertz region, which poses a challenge to the acquisition of the 4 × 4 MM. In this circumstance, the 3 × 3 MM degradation using only linear polarizations (LP) is preferred and sufficient for characterization of non-depolarizing samples. In this paper, a multi-spectral 3 × 3 MMP system based on the THz time-domain spectroscopy (THz-TDS) is established from 0.1 to 1 THz. The system demonstrated is capable of fulfilling the accurate determination of the 3 × 3 MM. The Mueller matrix polar decomposition (MMPD), modified to be compatible with the MM degradation, is employed to explore the fine details and properties of the sample. By signal post-processing techniques, the MM elements in the time domain are retrieved, and the time dimension reflecting the depth information facilitates the 3D reconstruction of the sample. This work provides a prototype for 3D imaging of biological samples at higher frequencies in the future.

Establishment of stable MRP1 knockdown by lentivirus-delivered shRNA in the mouse testis Sertoli TM4 cell line.

Sertoli cells around germ cells are considered a barrier that protects spermatogenesis from harmful influences. The transporter multidrug-resistance-associated protein 1 (MRP1) is a xenobiotic efflux pump that can export glutathione S-conjugated metabolites and xenobiotics from cells. In this study, the Mrp1 gene was stably knocked down in a mouse Sertoli cell line (TM4) using lentivirus vector-mediated RNA interference (RNAi) technology. Four shRNA interference sequences were chosen and designed to screen for the most effective shRNA in candidate cells. The results indicate that lentivirus vectors with high titres were generated and successfully transfected into TM4 cells with high efficiency. Puromycin was added to the culture medium to maintain constant selection during the establishment of the stable cell lines. The expression levels of Mrp1 mRNA and MRP1 protein in stably transfected TM4 cells were significantly lower than those in the control group. Importantly, the transport activity of MRP1 to Calcein and 5-carboxyseminaptharhodafluor (SNARF-1) were significantly reduced because of MRP1 silencing. Moreover, the silencing of the Mrp1 gene in the transfected TM4 cell lines remained highly stable for more than 6 months. These results suggest that the lentivirus-based RNAi stably knocks down the expression of the Mrp1 gene in the established TM4 cell line. This transfected TM4 cell line will provide a new and powerful tool to study the underlying mechanism of MRP1-mediated drug resistance and detoxication in the reproductive system.

Differentially expressed long non-coding RNAs and mRNAs of cadmium exposure on learning disability of offspring rats.

BACKGROUND: Cadmium (Cd) exposure has been found to have detrimental effects on the development of the central nervous system and cognitive ability in children. However, there is ongoing debate regarding the impact of maternal Cd exposure on the cognitive ability of offspring. In this study, we aimed to investigate the mechanisms underlying the influence of maternal Cd exposure on the cognitive ability of offspring rats. METHODS: Here, we constructed a model of cadmium poisoning in first-generation rats through gavage. The cognitive and memory abilities of its offspring were evaluated by water maze experiment. Then, we used the gene chip to find out the key genes, and we performed qRT-PCR detection of these genes. Subsequently, enrichment analysis was employed to identify pathways. Finally, we constructed a co-expression network consisting of LncRNAs and mRNAs to elucidate the biological functions and regulatory mechanisms of LncRNAs. RESULTS: The results of the water maze trial demonstrated that the offspring of rats exposed to cadmium in the first generation had reduced cognitive and memory abilities. Through an analysis of gene expression in the hippocampus of the cadmium-treated rats' offspring and the control group, we identified a correlation between the islet secretion pathway and the cognitive impairment observed in the offspring. Utilizing various algorithms, we identified Cpa1 and Prss1 as potential key genes associated with the cognitive impairment caused by cadmium. The results of qRT-PCR demonstrated a decrease in the expression levels of these genes in the hippocampus of the cadmium-treated rats' offspring. In addition, in the co-expression network, we observed that Cpa1 was co-expressed with 11 LncRNAs, while Prss1 was associated with 4 unexplored LncRNAs. Furthermore, we conducted an analysis to examine the relationship between Cpa1, Prss1-related transcription factors, and LncRNAs. CONCLUSION: Overall, this study provides novel insights into the molecular effects of first generation Cd exposure on the cognitive ability of offspring. The target genes and signaling pathways investigated in this study could serve as potential targets for improving neurodevelopment and cognitive ability in children.

Early plasma proteomic biomarkers and prediction model of acute respiratory distress syndrome after cardiopulmonary bypass: a prospective nested cohort study.

BACKGROUND: Early recognition of the risk of acute respiratory distress syndrome (ARDS) after cardiopulmonary bypass (CPB) may improve clinical outcomes. The main objective of this study was to identify proteomic biomarkers and develop an early prediction model for CPB-ARDS. METHODS: The authors conducted three prospective nested cohort studies of all consecutive patients undergoing cardiac surgery with CPB at Union Hospital of Tongji Medical College Hospital. Plasma proteomic profiling was performed in ARDS patients and matched controls (Cohort 1, April 2021-July 2021) at multiple timepoints: before CPB (T1), at the end of CPB (T2), and 24 h after CPB (T3). Then, for Cohort 2 (August 2021-July 2022), biomarker expression was measured and verified in the plasma. Furthermore, lung ischemia/reperfusion injury (LIRI) models and sham-operation were established in 50 rats to explore the tissue-level expression of biomarkers identified in the aforementioned clinical cohort. Subsequently, a machine learning-based prediction model incorporating protein and clinical predictors from Cohort 2 for CPB-ARDS was developed and internally validated. Model performance was externally validated on Cohort 3 (January 2023-March 2023). RESULTS: A total of 709 proteins were identified, with 9, 29, and 35 altered proteins between ARDS cases and controls at T1, T2, and T3, respectively, in Cohort 1. Following quantitative verification of several predictive proteins in Cohort 2, higher levels of thioredoxin domain containing 5 (TXNDC5), cathepsin L (CTSL), and NPC intracellular cholesterol transporter 2 (NPC2) at T2 were observed in CPB-ARDS patients. A dynamic online predictive nomogram was developed based on three proteins (TXNDC5, CTSL, and NPC2) and two clinical risk factors (CPB time and massive blood transfusion), with excellent performance (precision: 83.33%, sensitivity: 93.33%, specificity: 61.16%, and F1 score: 85.05%). The mean area under the receiver operating characteristics curve (AUC) of the model after 10-fold cross-validation was 0.839 (95% CI: 0.824-0.855). Model discrimination and calibration were maintained during external validation dataset testing, with an AUC of 0.820 (95% CI: 0.685-0.955) and a Brier Score of 0.177 (95% CI: 0.147-0.206). Moreover, the considerably overexpressed TXNDC5 and CTSL proteins identified in the plasma of patients with CPB-ARDS, exhibited a significant upregulation in the lung tissue of LIRI rats. CONCLUSIONS: This study identified several novel predictive biomarkers, developed and validated a practical prediction tool using biomarker and clinical factor combinations for individual prediction of CPB-ARDS risk. Assessing the plasma TXNDC5, CTSL, and NPC2 levels might identify patients who warrant closer follow-up and intensified therapy for ARDS prevention following major surgery.

Synthesis of Ester-Substituted Indolo[2,1-a]isoquinolines via Photocatalyzed Alkoxycarbonylation/Cyclization Reactions.

A direct alkoxycarbonylation/cyclization reaction is accomplished under visible light-induced photoredox catalysis. With this approach, a variety of ester-substituted indolo[2,1-a]isoquinolines are prepared in good to excellent yields. It is worth noting that this method not only can afford the synthesis of indolo[2,1-a]isoquinolines but also can provide an alternative route for generating complex target structures bearing carboxylic esters.

Spinal Cord Injury Inhibits the Differentiation and Maturation of NG2 Cells in the Cerebellum in Mice.

OBJECTIVE: Neuroimaging studies have shown that spinal cord injury (SCI) may lead to significant brain changes that are the key factors affecting functional recovery. However, little is known about the molecular and cellular biological mechanisms of these brain changes. The aim of this study was to investigate the molecular and cellular biological changes in the cerebellum after SCI. METHODS: A total of 72 mice were randomly divided into 2 groups: sham group and SCI group. A mouse model of SCI was established by an aneurysm clip. Pathological examinations of the injured site were performed by hematoxylin and eosin staining and immunohistochemical. Western blot and immunohistochemical were used to determine the effect of SCI on the differentiation and maturation of NG2 cells. RESULTS: Compared with the sham group, the spinal cord tissue structure was disrupted and the motor function decreased significantly in the SCI group; the number of NG2 cells in the ansiform lobule crus Ⅰ increased on the 7th and 14th days, whereas the expression of oligodendrocyte transcription factor 2, myelin basic protein, and proteolipid protein decreased on the 7th and 14th days after SCI. These results showed that the differentiation and maturation of NG2 cells in the ansiform lobule crus Ⅰ were inhibited after SCI, resulting in the decrease of the formation of mature oligodendrocytes. CONCLUSIONS: These results indicate that SCI can lead to secondary changes in the cerebellum, which may affect the functional recovery. These findings may be used as biomarkers to evaluate the secondary changes in the brain after SCI.

Identification of a apoptosis-related LncRNA signature to improve prognosis prediction and immunotherapy response in lung adenocarcinoma patients.

Apoptosis is closely associated with the development of various cancers, including lung adenocarcinoma (LUAD). However, the prognostic value of apoptosis-related lncRNAs (ApoRLs) in LUAD has not been fully elucidated. In the present study, we screened 2, 960 ApoRLs by constructing a co-expression network of mRNAs-lncRNAs associated with apoptosis, and identified 421 ApoRLs that were differentially expressed between LUAD samples and normal lung samples. Sixteen differentially expressed apoptosis-related lncRNAs (DE-ApoRLs) with prognostic relevance to LUAD patients were screened using univariate Cox regression analysis. An apoptosis-related lncRNA signature (ApoRLSig ) containing 10 ApoRLs was constructed by applying the Least Absolute Shrinkage and Selection Operator (LASSO) Cox regression method, and all LUAD patients in the TCGA cohort were divided into high or low risk groups. Moreover, patients in the high-risk group had a worse prognosis (p < 0.05). When analyzed in conjunction with clinical features, we found ApoRLSig to be an independent predictor of LUAD patients and established a prognostic nomogram combining ApoRLSig and clinical features. Gene set enrichment analysis (GSEA) revealed that ApoRLSig is involved in many malignancy-associated immunomodulatory pathways. In addition, there were significant differences in the immune microenvironment and immune cells between the high-risk and low-risk groups. Further analysis revealed that the expression levels of most immune checkpoint genes (ICGs) were higher in the high-risk group, which suggested that the immunotherapy effect was better in the high-risk group than in the low-risk group. And we found that the high-risk group was also better than the low-risk group in terms of chemotherapy effect. In conclusion, we successfully constructed an ApoRLSig which could predict the prognosis of LUAD patients and provide a novel strategy for the antitumor treatment of LUAD patients.

Discharge of pharmaceuticals from a municipal solid waste transfer station: Overlooked influence on the contamination of pharmaceuticals in surface waters.

During the temporal storage of municipal solid wastes (MSWs), pharmaceutically contained in MSWs may percolate into leachates and migrate into receiving waters via surface runoff. However, knowledge of their intra-event variations during the rainfall is quite limited. To fill in this gap, we collected runoff samples in a typical MSW transfer station over the full length of a rainfall event to comprehensively characterize the pharmaceutical contamination profiles. The results showed that 18 pharmaceuticals were detected in the runoff samples with high frequencies and concentrations ranging from below MQL to 18.6 µg/L. During the rainfall event, pharmaceuticals exhibited discrepant leachabilities as a result of different sorption capacities; two concentration peaks of each pharmaceutical were observed, suggesting the leaching effect by rainwater and the potential influence of human-related rinse. A further sampling campaign for one-week-long runoff samples generated by diurnal rinse water was conducted, and the results indicated comparable mass loads of pharmaceuticals in surface runoff receiving rinse water (0.37-8250 µg) to those in rainfall runoff (0.58-1754 µg), suggesting the similar discharge of pharmaceuticals from MSW transfer stations despite the weather. The estimated per capita discharge load of caffeine, one of the typical Pharmaceuticals, from MSW transfer stations was 4383 ng capita-1 d-1, higher than that in other emission sources, e.g. municipal wastewater effluent, indicating an overlooked influence of MSW transfer stations on its contamination in the surface waters in Shanghai.

Histomorphological and ultrastructural cadmium-induced kidney injuries and precancerous lesions in rats and screening for biomarkers.

Long-term exposure to cadmium (Cd) can severely damage the kidney, where orally absorbed Cd accumulates. However, the molecular mechanisms of Cd-induced kidney damage, especially the early biomarkers of Cd-induced renal carcinogenesis, are unclear. In the present study, we established a rat kidney injury model by intragastric administration of Cd to evaluate the morphological and biochemical aspects of kidney injury. We randomly divided Sprague-Dawley rats into control, low Cd (3 mg/kg), and high Cd (6 mg/kg) groups and measured biochemical indices associated with renal toxicity after 2, 4, and 8 weeks of treatment. The Cd-exposed mice had significantly higher Cd concentrations in blood and renal tissues as well as blood urea nitrogen (BUN), ß2-microglobulin (ß2-MG), urinary protein excretion, and tumor necrosis factor-α (TNF-α) levels. Furthermore, histopathological and transmission electron microscopy (TEM) observations revealed structural disruption of renal tubules and glomeruli after 8 weeks of exposure to the high Cd regimen. Besides, microarray technology experiments showed that Cd increased the expression of genes related to the chemical carcinogenesis pathway in kidney tissue. Finally, combining the protein-protein interaction (PPI) network of the Cd carcinogenesis pathway genes with the microarray and Comparative Toxicogenomics Database (CTD) results revealed two overlapping genes, CYP1B1 and UGT2B. Therefore, the combined molecular and bioinformatics experiments' results suggest that CYP1B1 and UGT2B are biomarkers of Cd-induced kidney injury with precancerous lesions.

Construction and evaluation of a nomogram for predicting survival in patients with lung cancer.

BACKGROUND: Lung cancer is a heterogeneous disease with a severe disease burden. Because the prognosis of patients with lung cancer varies, it is critical to identify effective biomarkers for prognosis prediction. METHODS: A total of 2325 lung cancer patients were integrated into four independent sets (training set, validation set I, II and III) after removing batch effects in our study. We applied the microarray data algorithm to screen the differentially expressed genes in the training set. The most robust markers for prognosis were identified using the LASSO-Cox regression model, which was then used to create a Cox model and nomogram. RESULTS: Through LASSO and multivariate Cox regression analysis, eight genes were identified as prognosis-associated hub genes, followed by the creation of prognosis-associated risk scores (PRS). The results of the Kaplan-Meier analysis in the three validation sets demonstrate the good predictive performance of PRS, with hazard ratios of 2.38 (95% confidence interval (CI), 1.61-3.53) in the validation set I, 1.35 (95% CI, 1.06-1.71) in the validation set II, and 2.71 (95% CI, 1.77-4.18) in the validation set III. Additionally, the PRS demonstrated superior survival prediction in subgroups by age, gender, p-stage, and histologic type (p < 0.0001). The complex model integrating PRS and clinical risk factors also have a good predictive performance for 3-year overall survival. CONCLUSIONS: In this study, we developed a PRS signature to help predict the survival of lung cancer. By combining it with clinical risk factors, a nomogram was established to quantify the individual risk assessments.

Distinct miRNAs associated with various clinical presentations of SARS-CoV-2 infection.

MicroRNAs (miRNAs) have been shown to play important roles in viral infections, but their associations with SARS-CoV-2 infection remain poorly understood. Here, we detected 85 differentially expressed miRNAs (DE-miRNAs) from 2,336 known and 361 novel miRNAs that were identified in 233 plasma samples from 61 healthy controls and 116 patients with COVID-19 using the high-throughput sequencing and computational analysis. These DE-miRNAs were associated with SASR-CoV-2 infection, disease severity, and viral persistence in the patients with COVID-19, respectively. Gene ontology and KEGG pathway analyses of the DE-miRNAs revealed their connections to viral infections, immune responses, and lung diseases. Finally, we established a machine learning model using the DE-miRNAs between various groups for classification of COVID-19 cases with different clinical presentations. Our findings may help understand the contribution of miRNAs to the pathogenesis of COVID-19 and identify potential biomarkers and molecular targets for diagnosis and treatment of SARS-CoV-2 infection.

Microarray analysis of differentially expressed long non-coding RNAs in daidzein-treated lung cancer cells.

Daidzein has been found to significantly inhibit the proliferation of lung cancer cells, while its potential molecular mechanisms remain unclear. To determine the molecular mechanism of daidzein on lung cancer cells, the Capital Bio Technology Human long non-coding (lnc) RNA Array v4, 4×180K chip was used to detect the gene expression profiles of 40,000 lncRNAs and 34,000 mRNAs in a human cancer cell line. Reverse transcription-quantitative (RT-q) PCR analysis was performed to detect the expression levels of target lncRNA and mRNAs in the H1299 cells treated with and without daidzein, using the lncRNA and mRNA gene chip. Bioinformatics analysis was performed to determine the differentially expressed genes from the results of the chip assays. There were 119 and 40 differentially expressed lncRNAs and mRNAs, respectively, that had a 2-fold change in expression level. A total of eight lncRNAs were upregulated in the H1299 lung cancer cells, while 111 lncRNAs were downregulated. Furthermore, five mRNAs were upregulated, and 35 mRNAs were downregulated. A total of six differentially expressed lncRNAs (ENST00000608897.1, ENST00000444196.1, ENST00000608741.1, XR_242163.1, ENST00000505196.1 and ENST00000498032.1) were randomly selected to validate the microarray data, which were consistent with the RT-qPCR analysis results. Differentially expressed mRNAs were enriched in important Gene Ontology terms and Kyoto Encyclopedia of Genes and Genomes pathways. Taken together, the results of the present study demonstrated that daidzein affected the expression level of lncRNAs in lung cancer cells, suggesting that daidzein may have potential effects on lung cancer cells.

Potential Correlation Between Depression-like Behavior and the Mitogen-Activated Protein Kinase Pathway in the Rat Hippocampus Following Spinal Cord Injury.

BACKGROUND: Depression induced by spinal cord injury (SCI) has been demonstrated in clinical and experimental studies; it significantly impacts patients' lives and may be associated with changes in the hippocampus. However, the biological mechanisms underlying depression after SCI are unknown. The mitogen-activated protein kinase (MAPK) signaling pathway participates in potential mechanisms of depression; it is unknown whether this pathway plays a role in SCI-induced depression. METHODS: We applied an animal model of depression induced by SCI, established using an aneurysm clip, to determine whether MAPK activation in the hippocampus is associated with depression-like behavior. RESULTS: SCI led to depression-like behavior, such as anhedonia in the sucrose preference test, decreased number of crossings in the open field test, decreased body weight, and decreased immobility time in the forced swim test. Western blot analysis further showed that SCI significantly increased the levels of phosphorylated p38 MAPK and cleaved caspase-3 in the hippocampus and inhibited the phosphorylation of extracellular signal-related kinase 1/2 and c-Jun N-terminal kinase 1/2. In addition, there were significant negative correlations between depression-like behavior and phosphorylated extracellular signal-related kinase 1/2 and positive correlations between depression-like behavior and phosphorylated p38 MAPK and cleaved caspase-3. CONCLUSIONS: These findings suggest that the MAPK pathway in the rat hippocampus may be involved in the pathophysiology of depression induced by SCI.

Development of an Immune-Related Risk Signature for Predicting Prognosis in Lung Squamous Cell Carcinoma.

Lung squamous cell carcinoma (LSCC) is the most common subtype of non-small cell lung cancer. Immunotherapy has become an effective treatment in recent years, while patients showed different responses to the current treatment. It is vital to identify the potential immunogenomic signatures to predict patient' prognosis. The expression profiles of LSCC patients with the clinical information were downloaded from TCGA database. Differentially expressed immune-related genes (IRGs) were extracted using edgeR algorithm, and functional enrichment analysis showed that these IRGs were primarily enriched in inflammatory- and immune-related processes. "Cytokine-cytokine receptor interaction" and "PI3K-AKT signaling pathway" were the most enriched KEGG pathways. 27 differentially expressed IRGs were significantly correlated with the overall survival (OS) of patients using univariate Cox regression analysis. A prognostic risk signature that comprises seven IRGs (GCCR, FGF8, CLEC4M, PTH, SLC10A2, NPPC, and FGF4) was developed with effective predictive performance by multivariable Cox stepwise regression analysis. Most importantly, the signature could be an independent prognostic predictor after adjusting for clinicopathological parameters, and also validated in two independent LSCC cohorts (GSE4573 and GSE17710). Potential molecular mechanisms and tumor immune landscape of these IRGs were investigated through computational biology. Analysis of tumor infiltrating lymphocytes and immune checkpoint molecules revealed distinct immune landscape in high- and low-risk group. The study was the first time to construct IRG-based immune signature in the recognition of disease progression and prognosis of LSCC patients.

Association of long-term sleep habits and hypertension: a cross-sectional study in Chinese adults.

Poor sleep has been associated with risk of hypertension, but previous studies were limited by treating one or two sleep factors as predictor. Our previous study has developed a sleep factor questionnaire (SFQ) to comprehensively assess wide range of sleep characteristics including sleep duration, sleep quality, light at night (LAN) exposure, night/shift work, daytime napping, and frequency of nighttime waking, insomnia, and snoring. In this cross-sectional study we used the SFQ to evaluated the associations between these sleep domains and hypertension. Comparing with the subjects who slept 7-8 h, subjects with sleep duration 6-7 h (OR = 1.42, 95% CI: 1.01, 2.00), 8-9 h (OR = 1.60, 95% CI: 1.07, 2.40), and over 9 h (OR = 2.39, 95% CI: 1.60, 3.58) had an increased prevalence of hypertension. The associations were significant among individuals under 45 years sleeping 6-7 h and those over 45 years sleeping over 8 h. The OR of prevalent hypertension was 0.80 (95% CI: 0.62, 0.97) for habitual daytime napping compared with never napping, and the association was also significant among individuals over 45 years old. Moreover, among subjects with sleep duration 7-8 h, habitual daytime napping may decrease prevalent hypertension (OR = 0.635, 95% CI: 0.437, 0.924). In conclusion, our data suggested a positive association between both short (6-7 h) and long sleep duration and hypertension, and a preventive pattern for habitual daytime napping among over 45 years old and hypertension. Moreover, interaction analysis indicated that habitual midday nap may decrease prevalent hypertension among subjects with sleep duration 7-8 h.

Maspin inhibits MCF-7 cell invasion and proliferation by downregulating miR-21 and increasing the expression of its target genes.

Maspin has been identified as a tumor suppressor gene in breast cancer, but the underlying regulatory mechanisms remain unclear. In the present study, maspin pcDNA was transfected into MCF-7 cells. microRNA (miR) microarray and reverse transcription-quantitative polymerase chain reaction was used for analysis; the results demonstrated that maspin may inhibit miR-10b, miR-21 and miR-451 expression in MCF-7 cells. In addition, maspin increased the expression of certain miR-21 target genes (phosphatase and tensin homolog, programmed cell death 4 and B-cell lymphoma-2), miR-10b target gene (Homeobox D10; HOXD10) and miR-451 target gene (multidrug resistance protein 1). Furthermore, the results of the present study revealed that decreased expression of miR-21 suppressed the invasion and proliferation of MCF-7 cells. Therefore, in the present study, it was hypothesized that as a tumor-suppressor gene, the potential molecular mechanism of maspin include down-regulating the expression of miR-21 and increasing the expression of specific miR-21 target genes.

Fluid Shear Stress Induces Endothelial Cell Injury via Protein Kinase C Alpha-Mediated Repression of p120-Catenin and Vascular Endothelial Cadherin In Vitro.

OBJECTIVE: The present study aimed to characterize the mechanism of fluid shear stress (FSS)-induced endothelial cell (EC) injury via protein kinase C alpha (PKCα)-mediated vascular endothelial cadherin (VE-cadherin) and p120-catenin (p120ctn) expression. METHODS: We designed a T chamber system that produced stable FSS on ECs in vitro. Human umbilical vein endothelial cells (HUVECs) in which PKCα was knocked down and normal HUVECs were cultured on the coverslips. FSS was impinged on these 2 types of ECs for 0 hours and 6 hours. The morphology and density of HUVECs were evaluated, and expression levels of phosphorylated PKCα, p120-catenin (p120ctn), VE-cadherin, phosphorylated p120ctn at S879 (p-S879p120ctn), and nuclear factor kappa B (NF-κB) were analyzed by Western blot. RESULTS: HUVECs exposed to FSS were characterized by a polygonal shape and decreased cell density. The phosphorylated PKCα level was increased under FSS at 6 hours (P < 0.05). In normal HUVECs during FSS, p120ctn and VE-cadherin were decreased, whereas p-S879p120ctn and NF-κB were increased, at 6 hours (P < 0.05). In HUVECs after PKCα knockdown, p120ctn and VE-cadherin were not significantly changed (P > 0.05), p-S879p120ctn was undetectable, but NF-κB was decreased (P < 0.05) at 6 hours. CONCLUSIONS: The possible mechanism of FSS-induced EC injury may be as follows: 1) PKCα induces low expression of p120ctn, which leads to activation of NF-κB and degradation of VE-cadherin; 2) PKCα-mediated phosphorylation of p120ctn at S879 disrupts p120ctn binding to VE-cadherin.

FOXC1-mediated LINC00301 facilitates tumor progression and triggers an immune-suppressing microenvironment in non-small cell lung cancer by regulating the HIF1α pathway.

BACKGROUND: Long non-coding RNAs (lncRNAs) are extensively intricate in the tumorigenesis and metastasis of various cancer types. Nevertheless, the detailed molecular mechanisms of lncRNA in non-small cell lung cancer (NSCLC) still remain mainly undetermined. METHODS: qPCR was performed to verify LINC00301 expression in NSCLC clinical specimens or cell lines. Fluorescence in situ hybridization (FISH) was conducted to identify the localization of LINC00301 in NSCLC cells. Chromatin immunoprecipitation (ChIP) was subjected to validate the binding activity between FOXC1 and LINC00301 promoters. RNA immunoprecipitation (RIP) was performed to explore the binding activity between LINC00301 and EZH2. RNA pull-down followed by dot-blot, protein domain mapping, and RNA electrophoresis mobility shift assay (EMSA) were conducted to identify the detailed binding regions between LINC00301 and EZH2. Alpha assay was conducted to quantitatively assess the interaction between LINC00301 and EZH2. RESULTS: LINC00301 is highly expressed in NSCLC and closely corelated to its prognosis by analyzing the relationship between differentially expressed lncRNAs and prognosis in NSCLC samples. in vitro and in vivo experiments revealed that LINC00301 facilitates cell proliferation, releases NSCLC cell cycle arrest, promotes cell migration and invasion, and suppresses cell apoptosis in NSCLC. In addition, LINC00301 increases regulatory T cell (Treg) while decreases CD8+ T cell population in LA-4/SLN-205-derived tumors through targeting TGF-ß. The transcription factor FOXC1 mediates LINC00301 expression in NSCLC. Bioinformatics prediction and in vitro experiments indicated that LINC00301 (83-123 nucleotide [nt]) can directly bind to the enhancer of zeste homolog 2 (EZH2) (612-727 amino acid [aa]) to promote H3K27me3 at the ELL protein-associated factor 2 (EAF2) promoter. EAF2 directly binds and stabilizes von Hippel-Lindau protein (pVHL), so downregulated EAF2 augments hypoxia-inducible factor 1 α (HIF1α) expression by regulating pVHL in NSCLC cells. Moreover, we also found that LINC00301 could function as a competing endogenous RNA (ceRNA) against miR-1276 to expedite HIF1α expression in the cytoplasm of NSCLC cells. CONCLUSIONS: In summary, our present research revealed the oncogenic roles of LINC00301 in clinical specimens as well as cellular and animal experiments, illustrating the potential roles and mechanisms of the FOXC1/LINC00301/EZH2/EAF2/pVHL/HIF1α and FOXC1/LINC00301/miR-1276/HIF1α pathways, which provides novel insights and potential theraputic targets to NSCLC.