Dehydroepiandrosterone attenuated the immune escape of oral squamous cell carcinoma through NF-κB p65/miR-15b-5p/B7-H4 axis.

OBJECTIVES: We aimed to explore the effects and mechanisms of action of dehydroepiandrosterone (DHEA) on immune evasion of oral squamous cell carcinoma (OSCC) to provide evidence for enhancing the effect of immunotherapy. MATERIALS AND METHODS: A xenograft mouse model and immunohistochemistry were used to reveal the patterns of tumor-infiltrating lymphocytes (TILs). The CAL27 and SCC VII cell lines were used for the in vitro study. Western blotting, qPCR, immunofluorescence, and flow cytometry were used to evaluate the expression of B7-H4. Recombinant mouse B7-H4 protein (rmB7-H4) and PG490, an inhibitor of NF-κB p65 were used for the "rescue study." Gain- and loss-of-function, luciferase reporter, and chromatin immunoprecipitation assays were performed to verify this mechanism. RESULTS: DHEA inhibited tumor growth in an OSCC xenograft mouse model, increased CD8 + cells, and decreased FOXP3 + cells in TILs. DHEA reduced the expression of B7-H4 in CAL27 and SCC VII cells RmB7-H4 reverses the effect of DHEA on tumor growth and TIL patterns. DHEA increased the expression of miR-15b-5p and activated its transcriptional factor NF-κB p65. Further experiments demonstrated that miR-15b-5p inhibited B7-H4 expression by binding to its 3'-UTR regions, and NF-κB p65 activated miR-15b transcription. PG490 reversed the effects of DHEA on tumor growth, antitumor immunity in the OSCC xenograft model, and the expression/phosphorylation of NF-κB p65, miR-15b-5p, and B7-H4. CONCLUSIONS: This study indicates that DHEA attenuates the immune escape of OSCC cells by inhibiting B7-H4 expression, providing new insights for cancer immunotherapy.

Genome-wide profiling of angiogenic cis-regulatory elements unravels cis-regulatory SNPs for vascular abnormality.

Angiogenesis is extensively involved in embryonic development and requires complex regulation networks, whose defects can cause a variety of vascular abnormalities. Cis-regulatory elements control gene expression at all developmental stages, but they have not been studied or profiled in angiogenesis yet. In this study, we exploited public DNase-seq and RNA-seq datasets from a VEGFA-stimulated in vitro angiogenic model, and carried out an integrated analysis of the transcriptome and chromatin accessibility across the entire process. Totally, we generated a bank of 47,125 angiogenic cis-regulatory elements with promoter (marker by H3K4me3) and/or enhancer (marker by H3K27ac) activities. Motif enrichment analysis revealed that these angiogenic cis-regulatory elements interacted preferentially with ETS family TFs. With this tool, we performed an association study using our WES data of TAPVC and identified rs199530718 as a cis-regulatory SNP associated with disease risk. Altogether, this study generated a genome-wide bank of angiogenic cis-regulatory elements and illustrated its utility in identifying novel cis-regulatory SNPs for TAPVC, expanding new horizons of angiogenesis as well as vascular abnormality genetics.

Intrarenal pressure study using 7.5 French flexible ureteroscope with or without ureteral access sheath in an ex-vivo porcine kidney model.

INTRODUCTION: 7.5F digital fURS and 9.5/11.5F ureteral access sheaths (UAS), both conventional (cUAS) and vacuum-assisted (vaUAS), are commercially available. Irrigation increases intrarenal pressure (IRP). This study analyzes the IRP with various irrigation rates using 7.5F fURS without UAS or with either cUAS or vaUAS in an ex-vivo porcine model. Pyelo-tubular backflow was also studied during these experiments. MATERIALS AND METHODS: 11 porcine kidneys were used. 7.5F digital fURS was tested without UAS and with 9.5/11.5F cUAS and vaUAS. 6F pressure monitor catheters were placed into the upper and lower calyces. IRPs were recorded under different irrigation rates. When vaUAS was used, the air vent was either open or closed. 300 mmHg aspiration pressure was chosen. Lastly, contrasted irrigation fluid was delivered until IRP reached above 30 mmHg. Fluoroscopy images were obtained at 5 mmHg intervals over this threshold to study the pyelo-tubular backflow. RESULTS: Using cUAS, IRP reached 30 mmHg with irrigation rates between 60 and 70 cc/min. Using vaUAS with vent closed, IRP never exceeded 10 mmHg with irrigation up to 120 cc/min. vaUAS with vent open performed marginally better than cUAS. fURS without UAS performed better than cUAS. Pyelo-tubular backflow became prominent at 40 mmHg. CONCLUSION: In an ex-vivo porcine model, 7.5F fURS could be used safely without UAS with irrigation rates up to 120 cc/min. The safety margin dropped to 60-70 cc/min with cUAS. vaUAS with vent closed maintained IRP < 10 mmHg with irrigation rates up to 120 cc/min. Pyelo-tubular backflow was observed with IRP > 35 mmHg.

Rapid identification and detection of aristolochic acids in the herbal extracts by Raman spectroscopy.

Herbs containing aristolochic acids (AAs) have already been proven to be highly carcinogenic and nephrotoxic. In this study, a novel surface-enhanced Raman scattering (SERS) identification method was developed. Ag-APS nanoparticles with a particle size of 3.53 ± 0.92 nm were produced by combining silver nitrate and 3-aminopropylsilatrane. The reaction between the carboxylic acid group of aristolochic acid I (AAI) and amine group of Ag-APS NPs was used to form amide bonds, and thus, concentrate AAI, rendering it easy to detect via SERS and amplified to obtain the best SERS enhancement effect. Detection limit was calculated to be approximately 40 nM. Using the SERS method, AAI was successfully detected in the samples of four Chinese herbal medicines containing AAI. Therefore, this method has a high potential to be applied in the future development of AAI analysis and rapid qualitative and quantitative analysis of AAI in dietary supplements and edible herbs.

Integrated genomic analysis identifies novel low-frequency cis-regulatory variant rs2279658 associated with VSD risk in Chinese children.

VSD combined with other cardiac or extracardiac malformations (defined as "complex VSD" by us) is one of the major causes of perinatal morbidity and mortality. Functional non-coding SNPs (cis-regulatory SNPs) have not been systematically studied in CHDs, including complex VSD. Here we report an exome-wide association analysis using WES data of 60 PA/VSD cases, 20 TOF cases and 100 controls in Chinese children. We identify 93 low-frequency non-coding SNPs associated with complex VSD risk. A functional genomics pipeline integrating ATAC-seq, ChIP-seq and promoter CHi-C recognizes the rs2279658 variant as a candidate cis-regulatory SNP. Specifically, rs2279658 resides in a cardiac-specific enhancer bound by FOXH1 and PITX2, and would abrogate binding of these two transcription factors to the identified enhancer during cardiac morphogenesis. COQ2 and FAM175A are predicted to be target genes for "rs2279658-FOXH1 or PITX2" pairs in the heart. These findings highlight the importance of cis-regulatory SNPs in the pathogenesis of complex VSD and broaden our understanding of this disease.

Optical electrocardiogram monitor with a real-time analysis of an abnormal heart rhythm for home-based medical alerts.

Sudden cardiac death (SCD) caused by cardiovascular disease is the greatest hidden danger to human life, accounting for about 25% of the total deaths in the world. Due to the early concealment of SCD and the heavy medical burden of long-term examination, telemedicine combined with home monitoring has become a potential medical alert method. Among all the existing human cardiac and electrophysiology monitoring methods, optics-based sensors attract the widest attention due to the advantages of low delay, real-time monitoring, and high signal-to-noise ratio. In this paper, we propose an optical sensor with the capabilities of long-term monitoring and real-time analysis. Combining an R-peak recognition algorithm, Lorenz plots (LP), and statistical analysis, we carried out the consistency analysis and result visualization of ECG sequences over 1 h. The results of 10 subjects show that the R-peak recognition accuracy of the optical ECG monitor is higher than 97.99%. The optical system can display abnormal heart rhythm in real-time through LP, and the readability is good, which makes the system suitable for self-monitoring at home. In addition, this paper provides a detailed long-term monitoring assessment method to effectively guide the practical clinical transformation of other optical wearable devices.

Comparison of intrarenal pressure between convention and vacuum-assisted ureteral access sheath using an ex vivo porcine kidney model.

OBJECTIVE: This study aimed to prove the vacuum-assisted ureteral access sheath (vaUAS) is more effective in maintaining a lower IRP than conventional ureteral access sheath (cUAS). MATERIALS: The model consisted of 12 freshly harvested adult porcine kidneys. METHODS: Either a 12/14F cUAS or vaUAS was alternately inserted into the ureter to one cm below the renal pelvis. Upper, middle, and lower calyces were punctured, and 6F pressure monitor catheters were introduced. IRP with cUAS was monitored using various irrigation rates. IRP with vaUAS was monitored with the same irrigation rates; various aspiration pressures; and vent fully closed, 50% closed, and fully open. RESULTS: cUAS with irrigation rate of 50 cc/min resulted in IRP < 30 mmHg. 50 to 100 cc/min should be used with caution. When irrigation rate exceeded 100 cc/min, IRP rose to ≥ 30 mmHg in most instances. With vent closed, vaUAS with vacuum pressure ≥ 150 mmHg and irrigation rate of 50 cc, 100 cc, and 150 cc/min generally resulted in IRPs < 5 mmHg. With vent half closed, vaUAS with vacuum pressure ≥ 300 mmHg and irrigation rate of ≤ 100 cc/min avoided IRP > 30 mmHg. vaUAS with vent open showed limited advantages over cUAS. CONCLUSION: vaUAS maintains lower IRP than cUAS under same parameters. Both vaUAS and cUAS can be used when irrigation is ≤ 50 cc/min vaUAS showed clear advantages over cUAS in maintaining lower pressure when irrigation rate is ≥ 100 cc/min.

KDELR1 Is an Independent Prognostic Predictor and Correlates With Immunity in Glioma.

Background: Gliomas are the most malignant central nervous system tumors. With the development of sequencing technology, more potential biomarkers related to the treatment, prognosis, and molecular classification of glioma have been identified. Here, we intend to investigate the potential biological function and clinical value of a new biomarker in glioma. Methods: KDELR1 expression data and the corresponding clinical information were downloaded from public databases and then preprocessed using R language. Correlation, Kaplan-Meier survival, and Cox regression analyses were performed to explore the clinical significance of KDELR1 in glioma patients. Furthermore, the immune infiltration and microenvironment parameters were evaluated via TIMER and CIBERSORT. Immunohistochemistry was conducted to confirm the KDELR1 expression and its correlation with immunity infiltration and prognosis. Results: KDELR1 was upregulated in glioma samples compared with normal brain tissues, and its expression was significantly correlated with age, the World Health Organization (WHO) grade, recurrence, necrosis, microvascular proliferation, molecular classification, isocitrate dehydrogenase (IDH) mutation, and 1p/19q codeletion status. In addition, survival analysis showed that glioma patients with KDELR1 overexpression had shorter overall survival (OS) and disease-free survival times, and Cox regression analysis revealed that KDELR1 acted as an independent prognostic factor of OS in glioma patients. Gene set enrichment analysis indicated a significant enrichment of metabolism-associated pathways. KDELR1 expression was positively associated with immune infiltration (including infiltration by CD8+ T cells, CD4+ T cells, macrophages, and so on) and microenvironment parameters (including stromal, immune, and ESTIMATE scores) in gliomas. The expression of KDELR1 and its correlation with the tumor grade and prognosis were confirmed by immunohistochemistry in clinical samples (n = 119, P < 0.05). Conclusions: Taken together, these findings suggest that KDELR1 is correlated with the tumor grade, molecular classifications, and immune infiltration; highlighting that KDELR1 is a novel and promising biomarker for molecular classification, treatment, and prognostic assessment may further indicate the treating effect of immune therapy.

Perspectives and Prospects on mRNA Vaccine Development for COVID-19.

BACKGROUND: The current coronavirus disease 2019 (COVID-19) pandemic, since first reported in Wuhan, has inspired worldwide efforts to develop effective COVID-19 vaccination strategies. mRNA vaccines encoding COVID-19 antigens have emerged prominantlyin this global race due to their high effectiveness and simple manufacturing process. Notably, two COVID-19 mRNA vaccines, mRNA-1273 and BNT162b2, have survived in clinical trials and been authorized for emergency use across variouscountries. SUMMARY: Recent advances on mRNA vaccine development for COVID-19 are discussed in this perspective, including sequence design, chemical modification, manufacturing process, and in vivo delivery. Phase I to IV clinical trials of mRNA-1273 and BNT162b2 are then summarized, respectively. CONCLUSION: Using mRNA vaccines is a promising strategy to achieve mass vaccination in the COVID-19 pandemic. We hope that future studies of mRNA vaccine technology will overcome existing limitations and help people cope with COVID-19.

Integrated weighted gene coexpression network analysis identifies Frizzled 2 (FZD2) as a key gene in invasive malignant pleomorphic adenoma.

BACKGROUND: Invasive malignant pleomorphic adenoma (IMPA) is a highly malignant neoplasm of the oral salivary glands with a poor prognosis and a considerable risk of recurrence. Many disease-causing genes of IMPA have been identified in recent decades (e.g., P53, PCNA and HMGA2), but many of these genes remain to be explored. Weighted gene coexpression network analysis (WGCNA) is a newly emerged algorithm that can cluster genes and form modules based on similar gene expression patterns. This study constructed a gene coexpression network of IMPA via WGCNA and then carried out multifaceted analysis to identify novel disease-causing genes. METHODS: RNA sequencing (RNA-seq) was performed for 10 pairs of IMPA and normal tissues to acquire the gene expression profiles. Differentially expressed genes (DEGs) were screened out with the cutoff criteria of |log2 Fold change (FC)|> 1 and adjusted p value < 0.05. Then, WGCNA was applied to systematically identify the hidden diagnostic hub genes of IMPA. RESULTS: In this research, a total of 1970 DEGs were screened out in IMPA tissues, including 1056 upregulated DEGs and 914 downregulated DEGs. Functional enrichment analysis was performed for identified DEGs and revealed an enrichment of tumor-associated GO terms and KEGG pathways. We used WGCNA to identify gene module most relevant with the histological grade of IMPA. The gene FZD2 was then recognized as the hub gene of the selected module with the highest module membership (MM) value and intramodule connectivity in protein-protein interaction (PPI) network. According to immunohistochemistry (IHC) staining, the expression level of FZD2 was higher in low-grade IMPA than in high-grade IMPA. CONCLUSION: FZD2 shows an expression dynamic that is negatively correlated with the clinical malignancy of IMPA and it plays a central role in the transcription network of IMPA. Thus, FZD2 serves as a promising histological indicator for the precise prediction of IMPA histological stages.

SYDE1 Acts as an Oncogene in Glioma and has Diagnostic and Prognostic Values.

Objectives: Gliomas remain one of serious public health problems worldwide which demand further and deeper investigation. The aim of this study was to explore the association between synapse defective protein 1 homolog 1 (SYDE1) and gliomas via public database analysis and in vitro validation to determine the potential diagnostic and prognostic values. Methods and Results: Compared with healthy brain tissues, there was a significant increase in SYDE1 expression in glioma tissues. Additionally, SYDE1 exhibited higher expression levels in glioma patients with unfavorable clinicopathological factors. In vitro knockdown of SYDE1 in glioma cell lines A172 inhibited their migrative and invasive ability but not the proliferative ability. GO and KEGG pathway analysis of the top 100 genes coexpressed with SYDE1 showed enrichments of tumor-associated terms. Further bioinformatic analysis revealed that the SNHG16/hsa-miR-520e/SYDE1 axis might be involved in glioma development. Conclusions: SYDE1 is expressed at higher levels in gliomas than in healthy brains, and can promote metastasis and invasion but not proliferation of gliomas. Furthermore, SYDE1 has values in the diagnosis and prognosis prediction of gliomas.

Identification of Expression Patterns and Potential Prognostic Significance of m5C-Related Regulators in Head and Neck Squamous Cell Carcinoma.

5-Methylcytosine (m5C) methylation is a major epigenetic technique of RNA modification and is dynamically mediated by m5C "writers," "erasers," and "readers." m5C RNA modification and its regulators are implicated in the onset and development of many tumors, but their roles in head and neck squamous cell carcinoma (HNSCC) have not yet been completely elucidated. In this study, we examined expression patterns of core m5C regulators in the publicly available HNSCC cohort via bioinformatic methods. The differentially expressed m5C regulators could divide the HNSCC cohort into four subgroups with distinct prognostic characteristics. Furthermore, a three-gene expression signature model, comprised of NSUN5, DNMT1, and DNMT3A, was established to identify individuals with a high or low risk of HNSCC. To explore the underlying mechanism in the prognosis of HNSCC, screening of differentially expressed genes, followed by the analysis of functional and pathway enrichment, from individuals with high- or low-risk HNSCC was performed. The results revealed a critical role for m5C RNA modification in two aspects of HNSCC: (1) dynamic m5C modification contributes to the regulation of HNSCC progression and (2) expression patterns of NSUN5, DNMT1, and DNMT3A help to predict the prognosis of HNSCC.

Comprehensive analysis of the transcriptome-wide m6A methylome in invasive malignant pleomorphic adenoma.

BACKGROUND: Invasive malignant pleomorphic adenoma (IMPA) is a highly invasive parotid gland tumor and lacks effective therapy. N6-Methyladenosine (m6A) is the most prevalent post-transcriptional modification of mRNAs in eukaryotes and plays an important role in the pathogenesis of multiple tumors. However, the significance of m6A-modified mRNAs in IMPA has not been elucidated to date. Hence, in this study, we attempted to profile the effect of IMPA in terms of m6A methylation in mRNA. METHODS: Methylated RNA immunoprecipitation with next-generation sequencing (MeRIP-seq) and RNA sequencing (RNA-seq) were utilized to acquire the first transcriptome-wide profiling of the m6A methylome map in IMPA followed by bioinformatics analysis. RESULTS: In this study, we obtained m6A methylation maps of IMPA samples and normal adjacent tissues through MeRIP-seq. In total, 25,490 m6A peaks associated with 13,735 genes were detected in the IMPA group, whereas 33,930 m6A peaks associated with 18,063 genes were detected in the control group. Peaks were primarily enriched within coding regions and near stop codons with AAACC and GGAC motifs. Moreover, functional enrichment analysis demonstrated that m6A-containing genes were significantly enriched in cancer and metabolism relevant pathways. Furthermore, we identified a relationship between the m6A methylome and the RNA transcriptome, indicating a mechanism by which m6A modulates gene expression. CONCLUSIONS: Our study is the first to provide comprehensive and transcriptome-wide profiles to determine the potential roles played by m6A methylation in IMPA. These results may open new avenues for in-depth research elucidating the m6A topology of IMPA and the molecular mechanisms governing the formation and progression of IMPA.

LncRNAs as Therapeutic Targets for Autophagy-involved Cardiovascular Diseases: A Review of Molecular Mechanism and T herapy Strategy.

BACKGROUND: Cardiovascular diseases (CVDs) remain the leading cause of death worldwide. The concept of precision medicine in CVD therapy today requires the incorporation of individual genetic and environmental variability to achieve personalized disease prevention and tailored treatment. Autophagy, an evolutionarily conserved intracellular degradation process, has been demonstrated to be essential in the pathogenesis of various CVDs. Nonetheless, there have been no effective treatments for autophagy- involved CVDs. Long noncoding RNAs (lncRNAs) are noncoding RNA sequences that play versatile roles in autophagy regulation, but much needs to be explored about the relationship between lncRNAs and autophagy-involved CVDs. SUMMARY: Increasing evidence has shown that lncRNAs contribute considerably to modulate autophagy in the context of CVDs. In this review, we first summarize the current knowledge of the role lncRNAs play in cardiovascular autophagy and autophagy-involved CVDs. Then, recent developments of antisense oligonucleotides (ASOs) designed to target lncRNAs to specifically modulate autophagy in diseased hearts and vessels are discussed, focusing primarily on structure-activity relationships of distinct chemical modifications and relevant clinical trials. PERSPECTIVE: ASOs are promising in cardiovascular drug innovation. We hope that future studies of lncRNA-based therapies would overcome existing technical limitations and help people who suffer from autophagy-involved CVDs.

GNG5 is an unfavourable independent prognostic indicator of gliomas.

Gliomas are the most common primary brain tumours, and glioblastomas (GBMs) are subgrouped into four distinct molecular subtypes. This study aimed to identify the potential gene related to glioma progression. Weighted gene co-expression network analysis (WGCNA) was used to explore the related gene. Correlation, ROC, survival and Cox regression analyses were performed. Blue module was strongly associated with WHO grade (r = .65, P = 1e-19). GNG5 in gliomas was overexpressed compared with normal samples and associated with clinicopathologic characteristics. GNG5 was frequent in Mesenchymal subtype and lowly expressed in Proneural subtype of GBMs. Survival and Cox regression analyses showed that glioma patients with GNG5 overexpression had shorter survival time, and GNG5 was an independent prognostic indicator of overall survival. Overall, GNG5 expression is closely associated with clinicopathologic characteristics and is an independent prognostic indicator for glioma patients, as well as a promising subtype-associated biomarker in molecular classification of gliomas.

Ultrabright single-band red upconversion luminescence in highly transparent fluorosilicate glass ceramics containing KMnF3 perovskite nanocrystals.

With a specially designed composition, highly transparent Yb3+/Er3+-doped fluorosilicate glass ceramic (GC) containing KMnF3 perovskite nanocrystals (NCs) is obtained for the first time. The rare-earth ions are preferentially accumulated in regions embedded with KMnF3 NCs; as a result, a remarkably enhanced (by an order of magnitude) single-band red upconversion luminescence (UCL) is achieved. Absolute quantum efficiency of the red UCL, which cannot be measured in previous GCs owing to insufficiency, reaches as high as 0.10%±0.02% in the GC sample reported in this Letter. This value is even higher than that of the well-known multiband emitting ß-NaYF4:Er3+/Yb3+ NCs and widely recognized GCs containing NaYF4:Yb3+/Er3+NCs.