Sequencing accuracy and systematic errors of nanopore direct RNA sequencing.

BACKGROUND: Direct RNA sequencing (dRNA-seq) on the Oxford Nanopore Technologies (ONT) platforms can produce reads covering up to full-length gene transcripts, while containing decipherable information about RNA base modifications and poly-A tail lengths. Although many published studies have been expanding the potential of dRNA-seq, its sequencing accuracy and error patterns remain understudied. RESULTS: We present the first comprehensive evaluation of sequencing accuracy and characterisation of systematic errors in dRNA-seq data from diverse organisms and synthetic in vitro transcribed RNAs. We found that for sequencing kits SQK-RNA001 and SQK-RNA002, the median read accuracy ranged from 87% to 92% across species, and deletions significantly outnumbered mismatches and insertions. Due to their high abundance in the transcriptome, heteropolymers and short homopolymers were the major contributors to the overall sequencing errors. We also observed systematic biases across all species at the levels of single nucleotides and motifs. In general, cytosine/uracil-rich regions were more likely to be erroneous than guanines and adenines. By examining raw signal data, we identified the underlying signal-level features potentially associated with the error patterns and their dependency on sequence contexts. While read quality scores can be used to approximate error rates at base and read levels, failure to detect DNA adapters may be a source of errors and data loss. By comparing distinct basecallers, we reason that some sequencing errors are attributable to signal insufficiency rather than algorithmic (basecalling) artefacts. Lastly, we generated dRNA-seq data using the latest SQK-RNA004 sequencing kit released at the end of 2023 and found that although the overall read accuracy increased, the systematic errors remain largely identical compared to the previous kits. CONCLUSIONS: As the first systematic investigation of dRNA-seq errors, this study offers a comprehensive overview of reproducible error patterns across diverse datasets, identifies potential signal-level insufficiency, and lays the foundation for error correction methods.

S100A12 is involved in the pathology of osteoarthritis by promoting M1 macrophage polarization via the NF-κB pathway.

BACKGROUND: Osteoarthritis (OA) is a degenerative joint disease that affects millions worldwide. Synovitis and macrophage polarization are important factors in the development of OA. However, the specific components of synovial fluid (SF) responsible for promoting macrophage polarization remain unclear. METHODS: Semi-quantitative antibody arrays were used to outline the proteome of SF. Differential expression analysis and GO/KEGG were performed on the obtained data. Immunohistochemistry and ELISA were used to investigate the relationship between SF S100A12 levels and synovitis levels in clinalclinical samples. In vitro cell experiments were conducted to investigate the effect of S100A12 on macrophage polarization. Public databases were utilized to predict and construct an S100A12-centered lncRNA-miRNA-mRNA competing endogenous RNA network, which was preliminarily validated using GEO datasets. RESULTS: The study outlines the protein profile in OA and non-OA SF. The results showed that the S100A12 level was significantly increased in OA SF and inflammatory chondrocytes. The OA synovium had more severe synovitis and higher levels of S100A12 than non-OA synovium. Exogenous S100A12 upregulated the levels of M1 markers and phosphorylated p65 and promoted p65 nuclear translocation, while pretreatment with BAY 11-7082 reversed these changes. It was also discovered that LINC00894 was upregulated in OA and significantly correlated with S100A12, potentially regulating S100A12 expression by acting as a miRNA sponge. CONCLUSIONS: This study demonstrated that S100A12 promotes M1 macrophage polarization through the NF-κB pathway, and found that LINC00894 has the potential to regulate the expression of S100A12 as a therapeutic approach.

Vibronic coupling of Rhodamine 6G molecules studied by doubly resonant sum frequency generation spectroscopy with narrowband infrared and broadband visible.

Doubly resonant sum frequency generation (DR-SFG) serves as a potent characteristic technique for probing the electronic spectra and vibronic coupling of molecules on surfaces. In this study, we successfully developed a novel infrared (IR)-white light (WL) DR-SFG spectroscopy based on narrowband IR and tunable broadband WL. This novel method was employed to explore the excitation spectrum and vibronic couplings of sub-monolayer Rhodamine 6G molecules. Our findings elucidate that the xanthene skeleton vibrational modes exhibit strong coupling with the S0-S1 electronic transition. Notably, we observed not only the 0-0 transition of the S0-S1 electronic continuum but also the 0-1 transition, a first time observation in the realm of DR-SFG spectroscopy. This advanced DR-SFG spectroscopy methodology facilitates a more sensitive examination of electronic spectra and the coupling between electronic transitions and vibrational modes, heralding a significant advancement in the understanding of molecular interactions on surfaces.

Surgical counting interruptions in operating rooms.

BACKGROUND: Operating rooms are complex working environments with high workloads and high levels of cognitive demand. The first surgical count which occurs during the chaotic preoperative stage and is considered a critical phase, is a routine task in ORs. Interruptions often occur during the first surgical count; however, little is known about the first surgical counting interruptions. This study aimed to observe and analyse the sources, outcomes, frequency of the first surgical counting interruptions and responses to interruptions. METHODS: A retrospective observational study was carried out to examine the occurrence of the first surgical counting interruptions between 1st August 2023 and 30th September 2023. The data were collected using the "Surgical Counting Interruption Event Form", which was developed by the researchers specifically for this study. RESULTS: A total of 66 circulating nurses (CNs) and scrub nurses (SNs) were observed across 1015 surgeries, with 4927.8 min of surgical count. The mean duration of the first surgical count was 4.85 min, with a range of 1.03 min to 9.51 min. In addition, 697 interruptions were identified, with full-term interruptions occurring an average of 8.7 times per hour. The most frequent source of interruption during the first surgical counts was instruments (N = 144, 20.7%). The first surgical counting interruptions mostly affected the CN (336 times; 48.2%), followed by the ORNs (including CNs and SNs) (243 times; 34.9%) and the SN (118 times; 16.9%). Most of the outcomes of interruptions were negative, and the majority of the nurses responded immediately to interruptions. CONCLUSIONS: The frequency of the first surgical counting interruption is high. Managers should develop interventions for interruptions based on different surgical specialties and different nursing roles.

Ideal topological Weyl complex phonons in two dimensions.

The advent of topological phonons has been attracting tremendous attention. However, studies in two-dimensional (2D) systems are limited. Here, we reveal a 2D novel combination of Weyl phonons - a Weyl complex composed of two linear Weyl nodes and one quadratic Weyl node. This Weyl complex consists of crossing points of two specific branches. We show that the coexistence of threefold symmetry - rotation symmetry, inversion symmetry, and time-reversal symmetry - could lead to the presence of the Weyl complex. Based on the symmetry requirement, we further construct the tight-binding model and effective k·p model for characterizing the Weyl complex. Moreover, due to the presence of the spacetime inversion symmetry, the linear and quadratic Weyl nodes feature a quantized (π and 2π) Berry phase, thus defining the corresponding topological charge. Furthermore, Weyl complexes consisting of Weyl points possess an emergent chiral symmetry, an integer topological charge is thus defined. Then, distinguished phenomena for the Weyl complex are studied, in particular, the edge states with three terminals. Our work predicts the presence of this novel 2D topological phase, and provides the symmetry guidance to realize it. Based on the first-principles calculations, we identify an existing material Cu2Si, as a concrete example to demonstrate the presence of the Weyl complex, and also study the phase transition under symmetry breaking.

A decision tree model of hypertension among college students in Yunnan Province, China.

PURPOSE: Previous studies have indicated that the prevalence rate of hypertension in adolescents is high, but it has not received much attention and the influencing factors are unclear, especially in Yunnan Province, China. MATERIALS AND METHODS: A cluster sampling method was used to investigate 4781 freshmen in a college in Kunming, Yunnan Province from November to December. Demographic and lifestyle data were collected using questionnaires, and height, weight and blood pressure were measured. Decision tree model of hypertension in college students was established by Chi-square automatic interactive detection method. RESULTS: Prevalence of prehypertension of systolic blood pressure (SBP) and diastolic blood pressure (DBP) were detected in 33.9% and 32.1%, respectively. Prevalence of hypertension of SBP and DBP was detected in 1.2% and 7.2%, respectively. The hypertension and prehypertension decision tree of SBP has gender (χ2 = 728.64, p < .001) at the first level and body mass index (BMI) (boys: χ2 = 55.98, p < .001; girls: χ2 = 79.58, p < .001) at the second level. The hypertension and prehypertension decision tree of DBP has gender (χ2 = 381.83, p < .001) at the first level, BMI (boys: χ2 = 40.54, p < .001; girls: χ2 = 48.79, p < .001) at the second level, only children (χ2 = 6.43, p = .04) and red wine consumption (χ2 = 8.17, p = .017) at the third level. CONCLUSIONS: The present study suggests that gender, BMI, only children and red wine consumption were the main factors affecting hypertension in college students in southwest border areas of China.

Hypertension in Chinese adolescent is generally ignored. This study first reports the prevalence of hypertension in adolescents in Yunnan Province, China.Four thousand seven hundred and eighty-one freshmen were surveyed and height, weight and blood pressure were measured. A decision tree model was used to analyze the predictors of hypertension.The study demonstrated that gender, BMI, only children and red wine predict hypertension in adolescents.

Efficacy of meniscus suture absorbability on meniscus healing success rate via second-look arthroscopy after meniscal repair: a systematic review and meta-analysis.

BACKGROUND: To preserve the meniscus's function, repairing the torn meniscus has become a common understanding. After which, the search for the ideal suture material is continuous. However, it is still controversial about the efficacy of suture absorbability on meniscus healing. METHODS: This review is designed according to the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) guidelines. INCLUSION CRITERIA: (1) Studies on meniscus repair; (2) Second-look arthroscopy was performed; (3) The meniscus was repaired by absorbable and non-absorbable sutures; (4) The healing condition of repaired meniscus via second-look arthroscopy was described. EXCLUSION CRITERIA: (1) Animal studies, cadaveric studies, or in vitro research; (2) Meniscus transplantation; (3) Open meniscus repair; (4) Reviews, meta-analysis, case reports, letters, and comments; (5) non-English studies. MEDLINE, Embase, and Cochrane Database were searched up to October 2022. Risk of bias and methodology quality of included literature were assessed according to ROBINS-I and the modified Coleman Methodological Scale (MCMS). Descriptive analysis was performed, and meta-analysis was completed by RevMan5.4.1. RESULTS: Four studies were included in the systematic review. Among them, three studies were brought into the meta-analysis, including 1 cohort study and 2 case series studies about 130 patients with meniscal tears combined with anterior cruciate ligament injury. Forty-two cases were repaired by absorbable sutures, and 88 were repaired by non-absorbable sutures. Using the fixed effect model, there was a statistical difference in the healing success rate between the absorbable and the non-absorbable groups [RR1.20, 95%CI (1.03, 1.40)]. CONCLUSION: In early and limited studies, insufficient evidence supports that non-absorbable sutures in meniscus repair surgery could improve meniscal healing success rate under second-look arthroscopy compared with absorbable sutures. In contrast, available data suggest that absorbable sutures have an advantage in meniscal healing. TRIAL REGISTRATION: The review was registered in the PROSPERO System Review International Pre-Registration System (Registration number CRD42021283739).

Exposure to Bisphenol A induces abnormal fetal heart development by promoting ferroptosis.

BACKGROUND: Bisphenol A (BPA), a common endocrine-disrupting chemical (EDC), has been revealed to be closely associated with the induction of abnormal heart development, obesity, prediabetes, and other metabolic disorders. However, the underlying mechanism of maternal BPA exposure on fetal heart development abnormalities is not clear. METHODS: To explore the adverse effects of BPA and its potential mechanism on heart development, C57BL/6 J mice and human cardiac AC-16 cells were used to conduct in vivo and in vitro studies. For the in vivo study, mice were exposed to low-dose BPA (40 mg/(kg·bw)) and high-dose BPA (120 mg/(kg·bw)) for 18 d during pregnancy. In vitro study, human cardiac AC-16 cells were exposed to BPA of various concentrations (0.01, 0.1, 1, 10, and 100 µM) for 24 h. Cell viability and ferroptosis were evaluated using 2,5-diphenyl-2 H-tetrazolium bromide (MTT), immunofluorescence staining, and western blotting. RESULTS: In BPA-treated mice, the alterations of fetal cardiac structure were observed. Increased NK2 homeobox 5(Nkx2.5) was detected in vivo with the induction of ferroptosis, revealing that BPA induced abnormal fetal heart development. Furthermore, the results showed that SLC7A11 and SLC3A2 decreased in low- and high-dose BPA-treated groups, suggesting that system Xc- mediated BPA-induced abnormal fetal heart development via inhibiting the expression of GPX4. Observing AC-16 cells confirmed that cell viability declined significantly with various concentrations of BPA. Moreover, BPA exposure suppressed GPX4 expression by inhibiting System Xc- (the decrease of SLC3A2 and SLC7A11). Collectively, system Xc- modulating cell ferroptosis might play important in abnormal fetal heart development induced by BPA exposure.

A Phase II Study of Pembrolizumab in Combination with Capecitabine and Oxaliplatin with Molecular Profiling in Patients with Advanced Biliary Tract Carcinoma.

BACKGROUND: We conducted a phase II study of the combination of pembrolizumab with capecitabine and oxaliplatin (CAPOX) in patients with advanced biliary tract carcinoma (BTC) to assess response rate and clinical efficacy. Exploratory objectives included correlative studies of immune marker expression, tumor evolution, and immune infiltration in response to treatment. PATIENTS AND METHODS: Adult patients with histologically confirmed BTC were enrolled and received oxaliplatin and pembrolizumab on day 1 of cycles 1-6. Capecitabine was administered orally twice daily as intermittent treatment, with the first dose on day 1 and the last dose on day 14 of cycles 1-6. Starting on cycle 7, pembrolizumab monotherapy was continued until disease progression. The primary endpoint was progression-free survival (PFS). Secondary endpoints were safety, tolerability, feasibility, and response rate. Immunohistochemistry (IHC) for PD-L1 and immune infiltrates was analyzed in paired tumor biopsies, as well as bulk transcriptome and exome profiling for five patients and single-cell RNA sequencing for one partial responder. RESULTS: Eleven patients enrolled, three of whom had received no prior systemic therapy. Treatment was well tolerated, and the most common treatment-related grade 3 or 4 adverse events were lymphocytopenia, anemia, and decreased platelet count. Three patients (27.3%) achieved a partial response, and six (54%) had stable disease. The disease control rate was 81.8%. The median PFS was 4.1 months with a 6-month PFS rate of 45.5%. Molecular profiling suggests qualitative differences in immune infiltration and clonal evolution based on response. CONCLUSION: Capecitabine and oxaliplatin in combination with pembrolizumab is tolerable and a potentially effective treatment for refractory advanced BTC. This study highlights a design framework for the precise characterization of individual BTC tumors. TRIAL REGISTRATION: This study was registered in ClinicalTrials.gov (NCT03111732).

Single-cell sequencing and its applications in bladder cancer.

Bladder cancer is the most common malignant tumour of the urinary system that is characterised by significant intra-tumoural heterogeneity. While large-scale sequencing projects have provided a preliminary understanding of tumour heterogeneity, these findings are based on the average signals obtained from the pooled populations of diverse cells. Recent advances in single-cell sequencing (SCS) technologies have been critical in this regard, opening up new ways of understanding the nuanced tumour biology by identifying distinct cellular subpopulations, dissecting the tumour microenvironment, and characterizing cellular genomic mutations. By integrating these novel insights, SCS technologies are expected to make powerful and meaningful changes to the current diagnosis and treatment of bladder cancer through the identification and usage of novel biomarkers as well as targeted therapeutics. SCS can discriminate complex heterogeneity in a large population of tumour cells and determine the key molecular properties that influence clinical outcomes. Here, we review the advances in single-cell technologies and discuss their applications in cancer research and clinical practice, with a specific focus on bladder cancer.

DeepViral: prediction of novel virus-host interactions from protein sequences and infectious disease phenotypes.

MOTIVATION: Infectious diseases caused by novel viruses have become a major public health concern. Rapid identification of virus-host interactions can reveal mechanistic insights into infectious diseases and shed light on potential treatments. Current computational prediction methods for novel viruses are based mainly on protein sequences. However, it is not clear to what extent other important features, such as the symptoms caused by the viruses, could contribute to a predictor. Disease phenotypes (i.e. signs and symptoms) are readily accessible from clinical diagnosis and we hypothesize that they may act as a potential proxy and an additional source of information for the underlying molecular interactions between the pathogens and hosts. RESULTS: We developed DeepViral, a deep learning based method that predicts protein-protein interactions (PPI) between humans and viruses. Motivated by the potential utility of infectious disease phenotypes, we first embedded human proteins and viruses in a shared space using their associated phenotypes and functions, supported by formalized background knowledge from biomedical ontologies. By jointly learning from protein sequences and phenotype features, DeepViral significantly improves over existing sequence-based methods for intra- and inter-species PPI prediction. AVAILABILITY AND IMPLEMENTATION: Code and datasets for reproduction and customization are available at https://github.com/bio-ontology-research-group/DeepViral. Prediction results for 14 virus families are available at https://doi.org/10.5281/zenodo.4429824. SUPPLEMENTARY INFORMATION: Supplementary data are available at Bioinformatics online.

MiR-145 is upregulated in the retarded preimplantation embryos and modulates cholesterol levels in mice preimplantation embryos through targeting Abca1.

BACKGROUND: Preimplantation embryonic lethality is a driver of female infertility. Certain microRNAs (miRNAs) have previously been demonstrated to play important roles in the regulation of embryogenesis. METHODS: Normally developing blastocysts and arrested embryos were collected from patients undergoing intracytoplasmic sperm injection (ICSI), and the expression of specific miRNAs therein was evaluated by qPCR. The overexpression of target molecule miR-145 in early mice embryos was achieved via oocyte microinjection, enabling the subsequent monitoring of how such overexpression impacted embryonic development. Bioinformatics approaches were utilized to identify putative miR-145 target mRNAs, and luciferase reporter assessments were implemented to confirm the ability of miR-145 to regulate Abca1 in HEK293T cells. The functional relationship between miR-145 and Abca1 in the mice's embryonic development was then confirmed through rescue assays. RESULTS: Abnormally increased miR-145 expression was observed in patients' arrested embryos, and the exogenous overexpression of this miRNA significantly suppressed mural blastocyst formation. Mechanistically, miR-145 was found to bind to the 3'-untranslated area of the Abca1 mRNA in HK293T cells, thus suppressing its expression and increasing embryonic cholesterol levels. In line with the importance of these cholesterol levels to embryogenesis, the upregulation of Abca1 was sufficient to rescue the observed change in cholesterol levels and the associated retardation of mice embryonic development that occurred in response to the overexpression of miR-145. CONCLUSION: The regulatory dynamics of the miR-145/Abca1 axis play an important role in shaping normal embryonic development.

CCL7 playing a dominant role in recruiting early OCPs to facilitate osteolysis at metastatic site of colorectal cancer.

BACKGROUND: Chemoattractant is critical to recruitment of osteoclast precursors and stimulates tumor bone metastasis. However, the role of chemoattractant in bone metastasis of colorectal cancer (CRC) is still unclear. METHODS: Histochemistry analysis and TRAP staining were utilized to detect the bone resorption and activation of osteoclasts (OCs) after administration of CCL7 neutralizing antibody or CCR1 siRNA. qRT-PCR analysis and ELISA assay were performed to detect the mRNA level and protein level of chemoattractant. BrdU assay and Tunel assay were used to detect the proliferation and apoptosis of osteoclast precursors (OCPs). The migration of OCPs was detected by Transwell assay. Western blots assay was performed to examine the protein levels of pathways regulating the expression of CCL7 or CCR1. RESULTS: OCPs-derived CCL7 was significantly upregulated in bone marrow after bone metastasis of CRC. Blockage of CCL7 efficiently prevented bone resorption. Administration of CCL7 promoted the migration of OCPs. Lactate promoted the expression of CCL7 through JNK pathway. In addition, CCR1 was the most important receptor of CCL7. CONCLUSION: Our study indicates the essential role of CCL7-CCR1 signaling for recruitment of OCPs in early bone metastasis of CRC. Targeting CCL7 or CCR1 could restore the bone volume, which could be a potential therapeutical target. Video Abstract.

Acute progressive large-area cerebral infarction caused by wasp sting: a case report.

Acute renal failure, multiple organ failure, and allergic reactions caused by wasp sting have been reported in many clinical reports, however, there are few reports on cerebral infarction due to wasp sting. We report a patient who developed acute progressive cerebral infarction after wasp stings, suffered from left-sided hemiplegia, clumsy speech, and left facial droop, only left the MRC grade 2 power and left conjugate gaze palsy after treatment. The MRA showed normal blood vessels. And the wasp sting was detected in head and right shoulder. We explored the mechanism by which the bee venom induced vasospasm and stimulated the superior cervical sympathetic ganglion leading to cerebral infarction. The treatment strategy is presented to provide experience for other clinicians.

Pathogenicity analysis and a novel case report of intronic mutations in CSF1R gene.

Colony-stimulating factor 1 receptor-associated leukoencephalopathy (CSF1R-related leukoencephalopathy) is a genetic disorder mutated in a single allele. It is characterized by an adult-onset along with predominantly cognitive impairment, accompanied by neuropsychiatric symptoms as well as motor symptoms such as Parkinsonism. In the current study, we confirmed a case of CSF1R-related leukoencephalopathy pedigree by genetic screening, and a new intron c. 1858 + 5 G > A mutation was detected in affected patients. After reviewing all previous reports of introns, we found that symptoms and clinical manifestations of the patients were typical and met the features of previous intron reports.

Value Of Anteroposterior-To-Transverse Ratio in Ultrasonic Diagnosis of Thyroid Nodule in Different Locations.

PURPOSE: To investigate the value of anteroposterior-to-transverse ratio (ATR) and the effect on features of nodules in ultrasound (US) diagnosis of thyroid nodules in different locations.  Methods: The nodules were divided into three groups according to the different nodule location: isthmus group; upper and lower poles of bilobed thyroid group; and the middle of the bilobed thyroid group. The diameters of the nodules were recorded, and ATR of the nodule was calculated on the transverse and longitudinal sections. The transverse and the longitudinal sections of ATR of thyroid nodules in different groups were compared.  Result: The transverse section of ATR was significantly different among the three groups (p = 0.001). In addition, there are significant differences in many US features among three groups, including nodule composition, thyroid parenchyma, morphology, echogenicity, shape, calcifications, vascularity, nodule ACR TI-RADS and histopathologic (all p < 0.05). In the group of upper and lower poles of bilobed thyroid, significant difference was found between the transverse and the longitudinal section of ATR (p = 0.019). The cut-off values of transverse section and longitudinal section of ATR were 0.967 and 0.750, respectively.  Conclusion: The transverse section of ATR at different location of thyroid may be a predictor for malignancy with clinical diagnostic significance.

A Novel Supervised Filter Feature Selection Method Based on Gaussian Probability Density for Fault Diagnosis of Permanent Magnet DC Motors.

For permanent magnet DC motors (PMDCMs), the amplitude of the current signals gradually decreases after the motor starts. In this work, the time domain features and time-frequency-domain features extracted from several successive segments of current signals make up a feature vector, which is adopted for fault diagnosis of PMDCMs. Many redundant features will lead to a decrease in diagnosis efficiency and increase the computation cost, so it is necessary to eliminate redundant features and features that have negative effects. This paper presents a novel supervised filter feature selection method for reducing data dimension by employing the Gaussian probability density function (GPDF) and named Gaussian vote feature selection (GVFS). To evaluate the effectiveness of the proposed GVFS, we compared it with the other five filter feature selection methods by utilizing the PMDCM's data. Additionally, Gaussian naive Bayes (GNB), k-nearest neighbor algorithm (k-NN), and support vector machine (SVM) are utilized for the construction of fault diagnosis models. Experimental results show that the proposed GVFS has a better diagnostic effect than the other five feature selection methods, and the average accuracy of fault diagnosis improves from 97.89% to 99.44%. This paper lays the foundation of fault diagnosis for PMDCMs and provides a novel filter feature selection method.

CircCAMSAP1 promotes hepatocellular carcinoma progression through miR-1294/GRAMD1A pathway.

Hepatocellular carcinoma (HCC) is one of the most common cancers with high prevalence and mortality, and it has brought huge economic and health burden for the world. It is urgent to found novel targets for HCC diagnosis and clinical intervention. Circular RNA (circRNA) has been reported to participate in many cancer progressions including HCC, suggesting that circRNA might paly essential role in HCC initiation and progression. Our study aims to found that potential circRNA participates in HCC development and its underlying molecular mechanisms. We obtained three pairs of HCC tissues and its adjacent normal tissues data from GEO DataSets. MTT, cell colony, EdU, wound-healing, transwell invasion and mouse xenograft model assays were used to demonstrate the biological functions of circCAMSAP1 in HCC progression. Furthermore, we conducted bioinformatics analysis, AGO2-RIP, RNA pull-down and luciferase reporter assays to assess the association of circCAMSAP1-miR-1294-GRAMD1A axis in HCC cells. The expression of circCAMSAP1 was up-regulated in HCC tissues compared with its adjacent normal tissues. Up-regulation of circCAMSAP1 promoted HCC biological functions both in vitro and in vivo. The promotive effects of circCAMSAP1 on HCC progression function through miR-1294/GRAMD1A pathway. CircCAMSAP1 was up-regulated in HCC tissues, and circCAMSAP1 up-regulated GRAMD1A expression to promote HCC proliferation, migration and invasion through miR-1294. CircCAMSAP1 might be a potential prognosis and therapeutic target for HCC.

Metformin alleviates the calcification of aortic valve interstitial cells through activating the PI3K/AKT pathway in an AMPK dependent way.

BACKGROUND: Calcific aortic valve disease (CAVD) is the most prevalent valvular disease worldwide. However, no effective treatment could delay or prevent the progression of the disease due to the poor understanding of its pathological mechanism. Many studies showed that metformin exerted beneficial effects on multiple cardiovascular diseases by mediating multiple proteins such as AMPK, NF-κB, and AKT. This study aims to verify whether metformin can inhibit aortic calcification through the PI3K/AKT signaling pathway. METHODS: We first analyzed four microarray datasets to screen differentially expressed genes (DEGs) and signaling pathways related to CAVD. Then aortic valve samples were used to verify selected genes and pathways through immunohistochemistry (IHC) and western blot (WB) assays. Aortic valve interstitial cells (AVICs) were isolated from non-calcific aortic valves and then cultured with phosphate medium (PM) with or without metformin to verify whether metformin can inhibit the osteogenic differentiation and calcification of AVICs. Finally, we used inhibitors and siRNA targeting AMPK, NF-κB, and AKT to study the mechanism of metformin. RESULTS: We screened 227 DEGs; NF-κB and PI3K/AKT signaling pathways were implicated in the pathological mechanism of CAVD. IHC and WB experiments showed decreased AMPK and AKT and increased Bax in calcific aortic valves. PM treatment significantly reduced AMPK and PI3K/AKT signaling pathways, promoted Bax/Bcl2 ratio, and induced AVICs calcification. Metformin treatment ameliorated AVICs calcification and apoptosis by activating the PI3K/AKT signaling pathway. AMPK activation and NF-κB inhibition could inhibit AVICs calcification induced by PM treatment; however, AMPK and AKT inhibition reversed the protective effect of metformin. CONCLUSIONS: This study, for the first time, demonstrates that metformin can inhibit AVICs in vitro calcification by activating the PI3K/AKT signaling pathway; this suggests that metformin may provide a potential target for the treatment of CAVD. And the PI3K/AKT signaling pathway emerges as an important regulatory axis in the pathological mechanism of CAVD.

Upregulated ox40l Can Be Inhibited by miR-146a-5p in Condylar Chondrocytes Induced by IL-1ß and TNF-α: A Possible Regulatory Mechanism in Osteoarthritis.

INTRODUCTION: Osteoarthritis (OA) is a common musculoskeletal disease characterized by pain, stiffness, limited activity, occasional effusion, and local inflammation. MiR-146 is one of the noncoding RNA closely related to OA, but the role of miR-146 in OA remains controversial. The tumour necrosis factor receptor OX40 is activated by its cognate ligand OX40L (TNFSF4) and functions as a T-cell costimulatory molecule. The T-cell functions, including cytokine production, expansion, and survival, are enhanced by the OX40 costimulatory signals. METHODS: We established an inflammatory model of condylar chondrocytes induced by IL-1ß and TNF-α and detected the expression of miRNA by miRNA sequencing. Then, cell transfection was used to study the role of miR146a-5p in OA. The Kyoto Encyclopedia of Genes and Genomes (KEGG) and database analysis were used to screen out potential target genes of miR-146a-5p. A dual luciferase activity assay tested whether ox40l is the target gene of miR-146a-5p. RESULTS: MiR-146a-5p and OX40L was upregulated after induced by IL-1ß and TNF-α, miR-146a-5p reduced the production of inflammatory factors but had no effect on chondrophenotypic factors, and ox40l was targeted by miR-146a-5p. CONCLUSION: OX40L and miR-146a-5p of condylar chondrocytes in the inflammatory environment (induced by IL-1ß and TNF-α) were significantly increased, miR-146a-5p is a protective factor in the inflammatory response, which can reduce the production of inflammatory factors, and miR-146a-5p may regulate T-cell-mediated immunity through targeting of ox40l in OA.