Identification of cytokines in benign and malignant thymus tumors: based on Mendelian randomization and proteomics.

Objective: The aim of this study was to identify potential causal cytokines in thymic malignancies and benign tumors from the FinnGen database using Mendelian randomization (MR). Methods: In this study, data from genome-wide association studies (GWAS) of 91 cytokines were used as exposure factors, and those of thymic malignant tumors and thymic benign tumors were the outcome variables. Two methods were used to determine the causal relationship between exposure factors and outcome variables: inverse variance weighting (IVW) and MR-Egger regression. Sensitivity analysis was performed using three methods, namely, the heterogeneity test, the pleiotropy test, and the leave-one-out test. Results: There was a causal relationship between the expression of fibroblast growth factor 5, which is a risk factor for thymic malignant tumors, and thymic malignant tumors. C-C motif chemokine 19 expression, T-cell surface glycoprotein CD5 levels, and interleukin-12 subunit beta levels were causally related to thymic malignant tumors and were protective. Adenosine deaminase levels, interleukin-10 receptor subunit beta expression, tumor necrosis factor (TNF)-related apoptosis-inducing ligand levels, and TNF-related activation-induced cytokine levels showed a causal relationship with thymic benign tumors, which are its risk factors. Caspase 8 levels, C-C motif chemokine 28 levels, interleukin-12 subunit beta levels, latency-associated peptide transforming growth factor beta 1 levels, and programmed cell death 1 ligand 1 expression showed a causal relationship with thymic benign tumors, which are protective factors. Sensitivity analysis showed no heterogeneity. Conclusion: Cytokines showed a causal relationship with benign and malignant thymic tumors. Interleukin-12 subunit beta is a common cytokine that affects malignant and benign thymic tumors.

Fish ubiquitin-specific protease 8 (USP8) inhibits IFN production through autophagy-lysosomal dependent degradation of IRF7.

Interferon regulatory factor 7 (IRF7) is considered the master regulator of virus-induced interferon (IFN) production. However, to avoid an autoimmune response, the expression of IRF7 must be tightly controlled. In this study, we report that zebrafish ubiquitin-specific protease 8 (USP8) promotes IRF7 degradation through an autophagy-lysosome-dependent pathway to inhibit IFN production. First, zebrafish usp8 is induced upon spring viremia of carp virus (SVCV) infection and polyinosinic/polycytidylic acid (poly I:C) stimulation. Second, overexpression of USP8 suppresses SVCV or poly I:C-mediated IFN expression. Mechanistically, USP8 interacts with IRF7 and promotes its degradation via an autophagy-lysosome-dependent pathway. Finally, USP8 significantly suppresses cellular antiviral responses and enhances SVCV proliferation. In summary, our discoveries offer a perspective on the role of zebrafish USP8 and provide additional understanding of the regulation of IRF7 in host antiviral immune response.

Achieving zero fouling in the ultrafiltration for secondary water supply systems in the absence of residual chlorine.

Ultrafiltration (UF) technology is widely used in secondary water supply systems (SWSS) to provide high-quality drinking water. However, the challenge of severe membrane fouling, which leads to frequent cleaning requirements, makes UF maintenance intensive. In this study, we tried to validate the feasibility of achieving zero fouling without the need for cleaning in the UF for SWSS, i.e., the fouling resistance can be maintained for a very long time without any increase. We operated dead-end UF systems at different fluxes, both with and without residual chlorine, and monitored the formation of fouling layers during filtration. The results demonstrated the successful achievement of zero fouling under a flux of 10 L/(m2 h) in the absence of chlorine, evidenced by no increase in transmembrane pressure for three months. This zero-fouling phenomenon was attributed to the formation of a self-regulating biofouling layer. This biofouling layer could degrade the deposited foulants and featured a loose morphology, facilitated by microbial activities in the cake layer. Although residual chlorine reduced the fouling rate by half at a flux of 30 L/(m2 h), it hindered the achievement of zero fouling at the lower flux of 10 L/(m2 h), due to its inhibitory effect on microbial activity. Intermittent operation of UF was effective in achieving zero fouling at higher fluxes (e.g., 30 L/(m2 h)). This benefit was primarily ascribed to the biodegradation of accumulated foulants and the expansion of biofouling layer during the pause of the intermittent filtration, which prompted the formation of biofouling layers with loose structure and balanced composition. To the best of our knowledge, this study is the first attempt to achieve zero fouling in UF for SWSS, and the findings may offer valuable insights for the development of cleaning-free and low-maintenance membrane processes.

Convenient synthesis and delivery of a megabase-scale designer accessory chromosome empower biosynthetic capacity.

Synthetic biology confers new functions to hosts by introducing exogenous genetic elements, yet rebuilding complex traits that are based on large-scale genetic information remains challenging. Here, we developed a CRISPR/Cas9-mediated haploidization method that bypasses the natural process of meiosis. Based on the programmed haploidization in yeast, we further developed an easy-to-use method designated HAnDy (Haploidization-based DNA Assembly and Delivery in yeast) that enables efficient assembly and delivery of large DNA, with no need for any fussy in vitro manipulations. Using HAnDy, a de novo designed 1.024 Mb synthetic accessory chromosome (synAC) encoding 542 exogenous genes was parallelly assembled and then directly transferred to six phylogenetically diverse yeasts. The synAC significantly promotes hosts' adaptations and increases the scope of the metabolic network, which allows the emergence of valuable compounds. Our approach should facilitate the assembly and delivery of large-scale DNA for expanding and deciphering complex biological functions.

YAP1/Piezo1 involve in the dynamic changes of lymphatic vessels in UVR-induced photoaging progress to squamous cell carcinoma.

BACKGROUND: UV-induced cutaneous squamous cell carcinoma (cSCC) is one of the most common skin cancers. The constant alterations of the lymphatic-centered immune microenvironment are essential in transforming from photoaging to cSCC. Studying the mechanism will be beneficial for new targets exploration to the early prediction of cSCC. AIMS: To investigate the dynamic changes and mechanism of the lymphatic-centered immune microenvironment in transforming from photoaging to cSCC induced by ultraviolet irradiation (UVR). METHODS: TIMER2.0 was used to analyze whether YAP1/VEGFC signaling pathway is involved in lymphangiogenesis in head and neck squamous cell carcinoma (HNSCC). Meanwhile, lymphatic-centered immune microenvironments alterations and the related cumulative survival time were also analyzed. With the accumulated UVR, skin photoaging developed and gradually progressed into actinic keratosis and cSCC on SKH-1 hairless mice. The skin lymphatic-centered immune microenvironment was evaluated at the 0th, 8th, 12th, 16-18th, and 20-24th week of UVR. Skin phenotype was assessed using optical coherence tomography (OCT) and skin image. H&E and Masson's trichrome staining evaluated epidermis and dermis. The structure of lymphatic vessels (LVs), blood vessels, and different types of T cells were evaluated by immunohistochemistry staining. The expression of Piezo1 whose deletion in adult lymphatics led to substantial valve degeneration, VE-cadherin that maintained the permeability of LVs, and YAP1 were evaluated by immunohistochemistry staining as well. Besides, the drainage function of LVs was assessed by Evans Blue assay in vivo. RESULTS: The lymphatic function and immune cell infiltration underwent adaptive changes under continuous UVR. TIMER2.0 analysis indicated that VEGFC genes high expressed in HNSCC. YAP1 gene expression was positive correlated with VEGFC in HNSCC. LV density increased in human cSCC. More LVs in HNSCC were beneficial to prolong the survival time. VEGFC gene overexpression was positive correlated to CD8+T cell infiltration. More CD8A+T cells and CD8B+T cell infiltration in HNSCC extended survival time. When YAP1 gene overexpression and high infiltration of endothelial cells took place simultaneously might prolong the survival time of HNSCC patients. And high infiltration of CD8+T cells prolonged the survival time as well. In animal studies, UVR-induced eight weeks (photoaging) and 16-18 weeks (precancerous) were two turning points. The density of LVs in UV-8w was the least. When photoaged skin developed into AK lesions (UV-16-18w), LV slightly exceeded healthy skin and proliferated sharply in cSCC (UV-20-24w). YAP1 expression was almost consistent with LV but rose after the photoaging stage. The drainage of cSCC mice induced by UVR was better than that of photoaged skin and worse than that of health skin. The dynamic alterations of LVs number, Piezo1 expression, and collagen might be reasons for it. The expression of Piezo1 was in the highest point after 8 weeks of UVR, then gradually descended to the platform. The total T cells increased slowly, but the infiltration of CD4+T cells increased, and CD8+T cells decreased after eight weeks of UVR. The CD8+T cells and CD4+T cells increased sharply in UV-16-18w and UV-20-24w groups. CONCLUSION: The lymphatic-centered immune microenvironment underwent adaptive changes under continuous UVR via regulating YAP1/VEGFC and Piezo1. During the formation of cSCC, there are two turning points, eight weeks (photoaging) and 16-18 weeks (precancerous). YAP1, Piezo1, LVs, and immune cells constantly changed with the skin state induced by UVR. According to these changes the process of cSCC can be identified in advance and intervene timely.

Evidence based on Mendelian randomization: Causal relationship between mitochondrial biological function and lung cancer and its subtypes.

OBJECTIVE: This study aimed to investigate the causal relationship between mitochondrial biological function and lung cancer, including its subtypes, via MR. METHODS: SNPs significantly associated with lung cancer and its subtypes were employed as instrumental variables. MR-Egger regression, simple mode, weighted mode, simple median, and weighted median, were utilized to determine the causal relationship between the exposure factor and the occurrence of lung cancer and its subtypes. RESULTS: NADH dehydrogenase (ubiquinone) flavoprotein 2 and transmembrane protein 70 were found to have a causal relationship with lung adenocarcinoma, acting as protective factors. The causal relationship between mitochondrial import inner membrane translocase subunit and NADH dehydrogenase (ubiquinone) iron-sulfur protein 4 and small-cell lung cancer was established as a risk factor. NADH dehydrogenase (ubiquinone) 1 beta subcomplex subunit 8 exhibited a causal relationship with small-cell lung cancer, acting as a protective factor. Furthermore, NAD-dependent protein deacylase sirtuin-5 was causally linked to lung squamous cell carcinoma, serving as a protective factor. A funnel plot demonstrated the symmetrical distribution of the SNPs. Thew pleiotroy test (P > 0.05) and "leave-one-out" test validated the relative stability of the results. CONCLUSION: This study established a causal relationship between mitochondrial biological function and lung cancer, including its subtypes.

Prognosis of recurrence after complete resection in early-stage lung adenocarcinoma based on molecular alterations: a systematic review and meta-analysis.

Molecular biomarkers have the potential to predict the recurrence risk of early-stage lung adenocarcinoma (LUAD) after complete resection, but the study results are controversial. We aimed to clarify the association of molecular alterations with disease-free survival (DFS) and recurrence-free survival (RFS) in early-stage LUAD with R0 resection. Comprehensive searches were conducted in PubMed/MEDLINE, Web of Science, and Cochrane Library for this systematic review and meta-analysis with date restrictions from 2012 to 2022. In the 18 included studies, data from a total of 7417 participants in 11 studies and 4167 participants in 9 studies were collected for the EGFR and KRAS meta-analyses, respectively. Two studies were assessed as having a moderate risk of bias, and the others were all assessed as having a high individual risk of bias. The molecular alterations in KRAS rather than EGFR, were associated with a high risk of recurrence for early-stage LUAD patients suffering from R0 resection, especially for those in pStage I, the pooled hazard ratios (HRs) of KRAS were 2.71 (95% CI, 1.81-4.06; I2 = 22%; P < 0.00001) and 1.95 (95% CI, 1.25-3.20; I2 = 57%; P = 0.003) with small interstudy heterogeneity in univariate and multivariate analyses, respectively. This finding suggests that molecular alterations in KRAS that could be detected by polymerase chain reaction techniques would provide new insight into stratifying risk and personalizing patient postoperative follow-up.

Using Fe(II)/Fe(VI) activated peracetic acid as pretreatment of ultrafiltration for secondary effluent treatment: Water quality improvement and membrane fouling mitigation.

Ultrafiltration (UF) is a technology commonly used to treat secondary effluents in wastewater reuse; however, it faces two main challenges: 1) membrane fouling and 2) inadequate nitrogen (N), phosphorus (P), and organic micropollutants (OMPs) removal. To address these two issues, in this study, we applied peracetic acid (PAA), Fe(VI)/PAA, and Fe(II)/PAA as UF pretreatments. The results showed that the most effective pretreatment was Fe(II)/200 µM PAA, which reduced the total fouling resistance by 90.2%. In comparison, the reduction was only 29.7% with 200 µM PAA alone and 64.3% with Fe(VI)/200 µM PAA. Fe(II)/200 µM PAA could effectively remove fluorescent components and hydrophobic organics in effluent organic matter (EfOM), and enhance the repulsive force between foulants and membrane (according to XDLVO analysis), and consequently, mitigate pore blocking and delay cake layer formation. Regarding pollutant removal, Fe(II)/200 µM PAA effectively degraded OMPs (>85%) and improved P removal by 58.2% via in-situ Fe(Ⅲ) co-precipitation. The quencher and probe experiments indicated that FeIVO2+, •OH, and CH3C(O)OO•/CH3C(O)O• all played important roles in micropollutant degradation with Fe(II)/PAA. Interestingly, PAA oxidation produced highly biodegradable products such as acetic acid, which significantly elevated the BOD5 level and increased the BOD5/total nitrogen (BOD5/TN) ratio from 0.8 to 8.6, benefiting N removal with subsequent denitrification. Overall, the Fe(II)/PAA process exhibits great potential as a UF pretreatment to control membrane fouling and improve water quality during secondary effluent treatment.

Sigma-1 receptor knockout disturbs gut microbiota, remodels serum metabolome, and exacerbates isoprenaline-induced heart failure.

Introduction: Heart failure (HF) is usually the end stage of the continuum of various cardiovascular diseases. However, the mechanism underlying the progression and development of HF remains poorly understood. The sigma-1 receptor (Sigmar1) is a non-opioid transmembrane receptor implicated in many diseases, including HF. However, the role of Sigmar1 in HF has not been fully elucidated. Methods: In this study, we used isoproterenol (ISO) to induce HF in wild-type (WT) and Sigmar1 knockout (Sigmar1-/-) mice. Multi-omic analysis, including microbiomics, metabolomics and transcriptomics, was employed to comprehensively evaluate the role of Sigmar1 in HF. Results: Compared with the WT-ISO group, Sigmar1-/- aggravated ISO-induced HF, including left ventricular systolic dysfunction and ventricular remodeling. Moreover, Sigmar1-/- exacerbated ISO-induced gut microbiota dysbiosis, which was demonstrated by the lower abundance of probiotics g_Akkermansia and g_norank_f_Muribaculaceae, and higher abundance of pathogenic g_norank_f_Oscillospiraceae and Allobaculum. Furthermore, differential metabolites among WT-Control, WT-ISO and Sigmar-/--ISO groups were mainly enriched in bile secretion, tryptophan metabolism and phenylalanine metabolism, which presented a close association with microbial dysbiosis. Corresponding with the exacerbation of the microbiome, the inflammation-related NOD-like receptor signaling pathway, NF-kappa B signaling pathway and TNF signaling pathway were activated in the heart tissues. Conclusion: Taken together, this study provides evidence that a Sigmar1 knockout disturbs the gut microbiota and remodels the serum metabolome, which may exacerbate HF by stimulating heart inflammation.

Identification of different types of tumors based on photoacoustic spectral analysis: preclinical feasibility studies on skin tumors.

Significance: Collagen and lipid are important components of tumor microenvironments (TME) and participates in tumor development and invasion. It has been reported that collagen and lipid can be used as a hallmark to diagnosis and differentiate tumors. Aim: We aim to introduce photoacoustic spectral analysis (PASA) method that can provide both the content and structure distribution of endogenous chromophores in biological tissues to characterize the tumor-related features for identifying different types of tumors. Approach: Ex vivo human tissues with suspected squamous cell carcinoma (SCC), suspected basal cell carcinoma (BCC), and normal tissue were used in this study. The relative lipid and collagen contents in the TME were assessed based on the PASA parameters and compared with histology. Support vector machine (SVM), one of the simplest machine learning tools, was applied for automatic skin cancer type detection. Results: The PASA results showed that the lipid and collagen levels of the tumors were significantly lower than those of the normal tissue, and there was a statistical difference between SCC and BCC (p<0.05), consistent with the histopathological results. The SVM-based categorization achieved diagnostic accuracies of 91.7% (normal), 93.3% (SCC), and 91.7% (BCC). Conclusions: We verified the potential use of collagen and lipid in the TME as biomarkers of tumor diversity and achieved accurate tumor classification based on the collagen and lipid content using PASA. The proposed method provides a new way to diagnose tumors.

Polarization Guided HDR Reconstruction via Pixel-Wise Depolarization.

Taking photos with digital cameras often accompanies saturated pixels due to their limited dynamic range, and it is far too ill-posed to restore them. Capturing multiple low dynamic range images with bracketed exposures can make the problem less ill-posed, however, it is prone to ghosting artifacts caused by spatial misalignment among images. A polarization camera can capture four spatially-aligned and temporally-synchronized polarized images with different polarizer angles in a single shot, which can be used for ghost-free high dynamic range (HDR) reconstruction. However, real-world scenarios are still challenging since existing polarization-based HDR reconstruction methods treat all pixels in the same manner and only utilize the spatially-variant exposures of the polarized images (without fully exploiting the degree of polarization (DoP) and the angle of polarization (AoP) of the incoming light to the sensor, which encode abundant structural and contextual information of the scene) to handle the problem still in an ill-posed manner. In this paper, we propose a pixel-wise depolarization strategy to solve the polarization guided HDR reconstruction problem, by classifying the pixels based on their levels of ill-posedness in HDR reconstruction procedure and applying different solutions to different classes. To utilize the strategy with better generalization ability and higher robustness, we propose a network-physics-hybrid polarization-based HDR reconstruction pipeline along with a neural network tailored to it, fully exploiting the DoP and AoP. Experimental results show that our approach achieves state-of-the-art performance on both synthetic and real-world images.

Hybrid High Dynamic Range Imaging fusing Neuromorphic and Conventional Images.

Reconstruction of high dynamic range image from a single low dynamic range image captured by a conventional RGB camera, which suffers from over- or under-exposure, is an ill-posed problem. In contrast, recent neuromorphic cameras like event camera and spike camera can record high dynamic range scenes in the form of intensity maps, but with much lower spatial resolution and no color information. In this article, we propose a hybrid imaging system (denoted as NeurImg) that captures and fuses the visual information from a neuromorphic camera and ordinary images from an RGB camera to reconstruct high-quality high dynamic range images and videos. The proposed NeurImg-HDR+ network consists of specially designed modules, which bridges the domain gaps on resolution, dynamic range, and color representation between two types of sensors and images to reconstruct high-resolution, high dynamic range images and videos. We capture a test dataset of hybrid signals on various HDR scenes using the hybrid camera, and analyze the advantages of the proposed fusing strategy by comparing it to state-of-the-art inverse tone mapping methods and merging two low dynamic range images approaches. Quantitative and qualitative experiments on both synthetic data and real-world scenarios demonstrate the effectiveness of the proposed hybrid high dynamic range imaging system. Code and dataset can be found at: https://github.com/hjynwa/NeurImg-HDR.

Comprehensive profiling of EGFR mutation subtypes reveals genomic-clinical associations in non-small-cell lung cancer patients on first-generation EGFR inhibitors.

Common sensitizing mutations in epidermal growth factor receptor (cEGFR), including exon 19 deletions (19-Del) and exon 21 L858R substitution, are associated with high sensitivity to EGFR-TKIs in NSCLC patients. The treatment for NSCLC patients with uncommon EGFR (uEGFR) mutations remains a subject of debate due to heterogeneity in treatment responses. In this manuscript, the targeted next-generation sequencing (NGS) data of a large cohort of EGFR-mutated NSCLC patients was assessed to elucidate genomic profiles of tumors carrying cEGFR or uEGFR mutations. The results showed that NSCLC patients with uEGFR mutations were more likely to harbor co-occurring genetic alterations in the Hippo pathway and a higher TMB compared with cEGFR-positive patients. Smoking-related mutations were found to significantly enriched in uEGFR-positive patients. Subgroup analyses were performed to identify potential prognostic biomarkers in patients harboring various EGFR subtype mutations. L858R-positive patients with co-existing ARID2 mutations had shorter progression-free survival (PFS) than those who were L858R- or 19-Del-positive but ARID2-negative (median: 2.3 vs. 12.0 vs. 8.0 months, P = 0.038). Furthermore, mutational profiles, such as top frequently mutated genes and mutational signatures of patients with various EGFR subtype mutations were significantly different. Our study analyzed the mutational landscape of NSCLC patients harboring cEGFR and uEGFR mutations, revealing specific genomic characteristics associated with uEGFR mutations that might explain the poor prognosis of first-generation EGFR-TKIs.

Numerical prediction of transient electrohydrodynamic instabilities under an alternating current electric field and unipolar injection.

In this paper, a direct numerical simulation (DNS) of dielectric fluid flow subjected to unipolar injection under an alternating current (AC) electric field is carried out. The effect of frequency f of pulsed direct current (PDC) and AC on the transient evolution of electroconvection and their subcritical bifurcations are investigated in details. Electroconvection under PDC or AC tends to exhibit oscillating flow due to the periodic boundary condition of charge density and potential compared to the direct current (DC) case. The results demonstrate that under the PDC field, the linear criterion T c decreases with increasing frequency, while the nonlinear stability criterion T f is hardly affected. Under the AC field, a critical frequency f c  = 0.0316 is found, which separates electroconvection into two typical flow regimes-periodic flow regime (f < f c ) and inhibited flow regime (f ≥ f c )-depending on whether free charges can reach the collector electrode before electric field inversion. AC-electrohydrodynamics (EHD) systems promote various flow patterns with relatively lower voltage regimes than DC-EHD systems. These mechanisms of electroconvection under the PDC/AC field offer unique possibilities for fluid flow control in biological EHD-driven flow and portable EHD applications.

Enhancing photoelectrochemical CO2 reduction with silicon photonic crystals.

The effectiveness of silicon (Si) and silicon-based materials in catalyzing photoelectrochemistry (PEC) CO2 reduction is limited by poor visible light absorption. In this study, we prepared two-dimensional (2D) silicon-based photonic crystals (SiPCs) with circular dielectric pillars arranged in a square array to amplify the absorption of light within the wavelength of approximately 450 nm. By investigating five sets of n + p SiPCs with varying dielectric pillar sizes and periodicity while maintaining consistent filling ratios, our findings showed improved photocurrent densities and a notable shift in product selectivity towards CH4 (around 25% Faradaic Efficiency). Additionally, we integrated platinum nanoparticles, which further enhanced the photocurrent without impacting the enhanced light absorption effect of SiPCs. These results not only validate the crucial role of SiPCs in enhancing light absorption and improving PEC performance but also suggest a promising approach towards efficient and selective PEC CO2 reduction.

Research on pharyngeal bacterial flora in transoral atlantoaxial operation and the postoperative follow-up study.

OBJECTIVES: To study the changes of bacterial flora after a series of preoperative oral disinfection and the postoperative recovery of nerve function of patients with craniovertebral junction disorders who were treated with transoral approach operations. METHODS: This research analyzed 20 cases collected from October 2009 to May 2010. All these patients were with CVJ disorders, including 8 males and 12 females, aged 2 to 66 (38.1 on average), and they were all treated with transoral approach operations. The mucosa samples of the posterior pharyngeal wall were sent for bacteria culture. These samples were collected by sterile cotton swabs at four crucial points, including 3 days before operation/before gargling, 3 days after continuous gargling/after anesthesia intubation on the day of operation, after intraoperative cleaning and washing of the mouth, and after intraoperative iodophor immersion. The microflora was stained by means of smear and further counted after an investigation by microscope. The neural function of patients was evaluated by the ASIA classification and the JOA scores. All patients but two with posterior stabilization performed respectively underwent transoral atlantoaxial reduction plate (TARP) fixation consecutively in the same sitting. A regular reexamination of cervical vertebra with lateral and open mouth X-ray, CT and MRI was conducted after operation to evaluate the reduction of atlantoaxial dislocation, internal fixation position, bone graft fusion, inflammatory lesions and tumor recurrence. RESULTS: This bacteriological research showed that the mucosa of the posterior pharyngeal wall of all the patients was in a sterile state after a series of oral preoperative preparations and intraoperative iodophor disinfection, which was considered as type I incision. The bacterial culture results of the mucosa samples of the posterior pharyngeal wall collected at different time points showed significant differences (χ2 = 42.762, P = 0.000). All the patients had improvement in ASIA, and their neural functions were improved to different levels after operation. There was a significant difference in JOA scores before and after operation (t = 8.677, P = 0.000). Postoperative imaging examination showed that the atlantoaxial screw position was good and firm, and the CVJ disorders were treated appropriately. CONCLUSION: It is safe and effective to cut the posterior pharyngeal muscle layer and implant internal fixation by means of transoral approach.

Identification of circRNAs involved in the development of hepatocellular carcinoma after insufficient radiofrequency ablation.

Previous studies have reported that circular RNAs (circRNAs) play a key role in the pathogenesis and progression of various diseases. In the present study, we aimed to identify potential circRNAs associated with the progression of hepatocellular carcinoma (HCC) after insufficient radiofrequency ablation (IRFA). A xenograft mouse IRFA model was initially established, and immunohistochemical staining (IHC) and polymerase chain reaction (PCR) were performed to confirm the expression of programmed cell death-ligand 1 (PD-L1) and vascular endothelial growth factor receptor-1 (VEGFR-1). CircRNA expression alterations were screened by next-generation sequencing (RNA-seq). Gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses were conducted to predict the function of genes coding differentially expressed circRNAs. The selected circRNAs were validated utilizing PCR and Sanger sequencing. The relationships between circRNAs, microRNAs, PD-L1, and VEGFR-1 were predicted by bioinformatics. Overall, a total of 612 circRNAs were differentially expressed in IRFA-treated subcutaneous tumorigenesis tissue. Among them, 435 circRNAs were significantly upregulated and 177 circRNAs were downregulated. GO and KEGG analyses were employed to predict the functions of these circRNAs. Thereafter, quantitative reverse transcription PCR (qRT-PCR) assays determined that these seven circRNAs were overexpressed in the IRFA group, which was consistent with the RNA-seq results. Based on the bioinformatic analysis, seven circRNAs confirmed by Sanger sequencing were predicted to likely regulate PD-L1 and VEGFR-1 expression levels by functioning as sponges for microRNAs (miRNAs) and forming a circRNA-miRNA-PD-L1/VEGFR-1 regulatory network. Finally, IHC and qRT-PCR of PD-L1 and VEGFR-1 confirmed the activation of this pathway. Taken together, we report that differentially expressed circRNAs might simultaneously regulate PD-L1 and VEGFR-1 in the IRFA tissues, which provides a novel view of circRNAs in HCC progression after the IRFA procedure.

Modified 5-aminolevulinic acid photodynamic therapy reduces pain and improves therapeutic effects in cutaneous squamous cell carcinoma mouse model.

BACKGROUND AND OBJECTIVES: Conventional ALA-PDT (C-PDT) has limited efficacy in cutaneous squamous cell carcinoma (cSCC), and there is obvious pain during treatment, which limits its clinical application. We sought to modify photodynamic therapy into a more painless and effective treatment. METHODS: We modified C-PDT by reducing the incubation time of the pro-sensitizer and increasing the light dose; we named this method modified ALA-PDT (M-PDT). We compared the pain response and curative effect between C-PDT and M-PDT in cSCC mouse models. Pain-related proteins were examined by western blot analysis and immunohistochemistry. Tumor progression-associated signaling pathways were analyzed by RNA-seq and western blot analysis. Reactive oxygen species (ROS) generation was measured with a ROS test kit and Microplate reader. RESULTS: M-PDT greatly reduced pain during treatment. Interestingly, when the cSCC tumor volume increased to 150-200 mm3 , M-PDT almost completely eliminated the tumors, while C-PDT did not. The better curative effect of M-PDT might be due to the stronger suppression of the Stat3, Erk1/2, and mTOR signaling pathways. Moreover, flow cytometry demonstrated that M-PDT could recruit CD8+ T cells to inhibit cSCC progression. Further investigation determined that the different mechanisms of C-PDT and M-PDT were related to more ROS generation induced by M-PDT. CONCLUSIONS: Our results suggest that M-PDT, which is more painless and effective than C-PDT, is expected to provide a solution for the treatment of cSCC.

Self-Enhanced False Memory Across the Life Span.

OBJECTIVES: The role of self in veridical memory has been extensively studied, but what is the role of self in false memory development across the life span? The current study examined the impact of self-reference on associative false memory in children, younger adults, and older adults, and further investigated possible mechanisms concerning how self-reference might affect false memory in different age groups. METHODS: Combining a self-reference manipulation with the Deese/Roediger-McDermott (DRM) paradigm, children, younger adults, and older adults encoded DRM word lists as paired with their own name, another person's name, or a red square. Later their true and false recognition memory as well as recollection and familiarity were measured. RESULTS: A self-enhanced false memory effect was found in all age groups. That is, participants generated more false memories in the self-reference condition relative to the other-reference and neutral conditions. Furthermore, when examining its underlying memory mechanisms, we found that self-reference mainly increased false recollection in younger adults but facilitated familiarity of critical lures in older adults. DISCUSSION: Although self-reference increases false memory in both younger and older adults, the underlying mechanisms are different in that older adults have more self-relevant false familiarity while younger adults generate more self-relevant phantom recollection. The current study also has implications for eyewitness reports, suggesting that the self-relevance of memory may be one relevant factor to consider when evaluating potential risk factors of false memory.

DNA N6-methyladenine involvement and regulation of hepatocellular carcinoma development.

DNA N6-methyladenine (6 mA) is a new type of DNA methylation identified in various eukaryotic cells. However, its alteration and genomic distribution features in hepatocellular carcinoma (HCC) remain elusive. In this study, we found that N6AMT1 overexpression increased HCC cell viability, suppressed apoptosis, and enhanced migration and invasion, whereas ALKBH1 overexpression induced the opposite effects. Further, 23,779 gain-of-6 mA regions and 11,240 loss-of-6 mA regions were differentially identified in HCC tissues. The differential gain and loss of 6 mA regions were considerably enriched in intergenic regions. Moreover, 7% of the differential 6 mA modifications were associated with tumors, with 60 associated with oncogenes and 57 with tumor suppressor genes (TSGs), and 17 were common to oncogenes and TSGs. The candidate genes affected by 6 mA were filtered by gene ontology (GO) and RNA-seq. Using quantitative polymerase chain reaction (qPCR), BCL2 and PARTICL were found to be correlated with DNA 6 mA in certain HCC processes.