Exploring Circular RNA Profile and Expression in Extracellular Vesicles.

Extracellular vesicles (EVs) are small vesicles secreted by various cell types and are enriched in multiple body fluids. EVs containing RNA have the potential to modulate biological processes and are being investigated for their diagnostic and therapeutic applications. Circular RNAs (circRNAs), generated through back-splicing of exons, are enriched in EVs. Given their unique characteristics and diverse functions, EV-circRNAs are important players in disease pathology. This chapter describes a workflow for investigating the expression profile of EV-circRNAs, which includes EVs separation, library preparation, and bioinformatics analysis. This workflow can aid the investigation of EV-circRNAs and their potential role in disease pathology.

Genomic and transcriptional characterization of early esophageal squamous cell carcinoma.

BACKGROUND: Esophageal squamous cell carcinoma (ESCC) is a highly heterogeneous cancer that lacks comprehensive understanding and effective treatment. Although multi-omics study has revealed features and underlying drivers of advanced ESCC, research on molecular characteristics of the early stage ESCC is quite limited. MATERIALS AND METHODS: We presented characteristics of genomics and transcriptomics in 10 matched pairs of tumor and normal tissues of early ESCC patients in the China region. RESULTS: We identified the specific patterns of cancer gene mutations and copy number variations. We also found a dramatic change in the transcriptome, with more than 4,000 genes upregulated in cancer. Among them, more than one-third of HOX family genes were specifically and highly expressed in early ESCC samples of China and validated by RT-qPCR. Gene regulation network analysis indicated that alteration of Hox family genes promoted the proliferation and metabolism remodeling of early ESCC. CONCLUSIONS: We characterized the genomic and transcriptomic landscape of 10 paired normal adjacent and early ESCC tissues in the China region, and provided a new perspective to understand the development of ESCC and insight into potential prevention and diagnostic targets for the management of early ESCC in China.

Plasma extracellular vesicle transcriptomics identifies CD160 for predicting immunochemotherapy efficacy in lung cancer.

Better biomarkers are needed to improve the efficacy of immune checkpoint inhibitors in lung adenocarcinoma (LUAD) treatment. We investigated the plasma extracellular vesicle (EV)-derived long RNAs (exLRs) in unresectable/advanced LUAD to explore biomarkers for immunochemotherapy. Seventy-four LUAD patients without targetable mutations receiving first-line anti-programmed cell death 1 (PD-1) immunochemotherapy were enrolled. Their exLRs were profiled through plasma EV transcriptome sequencing. Biomarkers were analyzed against response rate and survival using pre- and post-treatment samples in the retrospective cohort (n = 36) and prospective cohort (n = 38). The results showed that LUAD patients demonstrated a distinct exLR profile from the healthy individuals (n = 56), and T-cell activation-related pathways were enriched in responders. Among T-cell activation exLRs, CD160 exhibited a strong correlation with survival. In the retrospective cohort, the high baseline EV-derived CD160 level correlated with prolonged progression-free survival (PFS) (P < 0.001) and overall survival (OS) (P = 0.005), with an area under the curve (AUC) of 0.784 for differentiating responders from non-responders. In the prospective cohort, the CD160-high patients also showed prolonged PFS (P = 0.003) and OS (P = 0.014) and a promising AUC of 0.648. The predictive value of CD160 expression was validated by real-time quantitative PCR. We also identified the dynamics of EV-derived CD160 for monitoring therapeutic response. The elevated baseline CD160 reflected a higher abundance of circulating NK cells and CD8+ -naïve T cells, suggesting more active host immunity. In addition, increased CD160 levels of tumors also correlated with a favorable prognosis in LUAD patients. Together, plasma EV transcriptome analysis revealed the role of the baseline CD160 level and early post-treatment CD160 dynamics for predicting the response to anti-PD-1 immunochemotherapy in LUAD patients.

circTMEM181 upregulates ARHGAP29 to inhibit hepatocellular carcinoma migration and invasion by sponging miR-519a-5p.

AIM: Circular RNAs (circRNAs) are a novel class of noncoding RNAs and are conserved in various species. Although numerous circRNAs have been identified, their role in cancer remains unclear. METHODS: The expression of circTMEM181 in 90 paired human hepatocellular carcinoma (HCC) and adjacent nontumor tissues were detected using quantitative reverse transcription-polymerase chain reaction. Transwell assay was performed for functional analysis of HCC cell migration and invasion. Luciferase reporter assay was used to verify the combination of circTMEM181 and miR-519a-5p. RESULTS: In this study, we identified a novel circRNA, named circTMEM181, was downregulated in HCC tissues. Decreased expression of circTMEM181 was associated with shorter overall survival of patients with HCC. CircTMEM181 overexpression reduced HCC cell migration and invasion abilities, while circTMEM181 knockdown increased cell motility. Mechanically, circTMEM181 could directly bind to miR-519a-5p and subsequently upregulate ARHGAP29 protein expression. CONCLUSION: These data provide the first evidence of clinical significance and function of circTMEM181, and suggest the circTMEM181/miR-519a-5p/ARHGAP29 axis in HCC development.

Sargassum fusiforme Fucoidan Alleviates High-Fat Diet-Induced Obesity and Insulin Resistance Associated with the Improvement of Hepatic Oxidative Stress and Gut Microbiota Profile.

Sargassum fusiforme fucoidan (SFF) exhibits diverse biological activities. Insulin resistance (IR) implicated in type 2 diabetes (T2D) has become an epidemic health issue worldwide. In this study, we investigated whether SFF can improve insulin sensitivity in high-fat diet (HFD)-fed mice. Our present data showed that SFF significantly reduced fasting blood glucose and IR index along with improved glucose tolerance. Impaired phosphorylation of Akt was also restored by SFF. Furthermore, SFF decreased the levels of MDA and 4-HNE-modified protein and increased GSH/GSSG ratio as well as elevated antioxidant enzymes and activated Nrf2 signaling. SFF also increased the abundance and diversity of gut microbiota in the obese mice, as well as improved intestinal integrity and inflammation. Our findings suggested that SFF ameliorated HFD-induced IR through activating the Nrf2 pathway, remodeling gut microbiota, and reducing intestinal inflammation, thus providing a novel perspective into the treatment strategy on metabolic disease.

[Design and application of the machine for cardiopulmonary resuscitation based on scissor lift principle without back plate].

Cardiopulmonary resuscitation (CPR) guidelines emphasize that external chest compression should be started as early as possible during CPR. In addition to manual hand compressions, the machine for CPR can be used instead. However traditional machines for CPR take a lot of time toinstall, and is not conducive to the early initiation of chest compressions. So the medical staff of the emergency department of Mindong Hospital Affiliated to Fujian Medical University designed a new type of machine for CPR based on scissor lift principle without back plate. The design has obtained the national utility model patent. This patented product with flexible lifting and accurate placement is easy to install and fix, and can overcome the drawbacks of conventional machines that are uneasy to install, thus ensuring the implementation of high quality CPR.

Different extraction methods bring about distinct physicochemical properties and antioxidant activities of Sargassum fusiforme fucoidans.

Fucoidan is a complex sulfated polysaccharide and an active component found in the cell wall of brown seaweeds. In the present study, fucoidans were obtained from Sargassum fusiforme using different extraction methods, including hot water (prepared fucoidan was named as WSFF), dilute hydrochloric acid (ASFF), and calcium chloride solution (CSFF). The assessments were performed on S. fusiforme fucoidans based on their chemical composition, molecular conformations, and in vitro antioxidant activities. ASFF showed the maximum extraction yield (11.24%), whereas CSFF exhibited the minimum yield (3.94%). The monosaccharide composition of WSFF, ASFF, and CSFF was similar, but the molar ratio of monosaccharide was quite different. Moreover, their molecular weight, Fourier transform infrared (FT-IR) spectrum, surface morphology, uronic acid content and degree of sulfation were distinct. The Congo red test and Circular dichroism spectroscopy analysis displayed some differences in solution conformation of these samples. Furthermore, WSFF, ASFF, and CSFF showed distinct in vitro antioxidant activities evaluated by DPPH and hydroxyl radical scavenging assays. The present study provides scientific evidence on the influences of extraction methods on the physicochemical characteristics, conformation behaviors and antioxidant activities of S. fusiforme fucoidans.