Construction of a Cancer Stem Cell related Histone Acetylation Regulatory Genes Prognostic Model for Hepatocellular Carcinoma via Bioinformatics Analysis: Implications for Tumor Chemotherapy and Immunity.

BACKGROUND: Cancer stem cells (CSC) play an important role in the development of Liver Hepatocellular Carcinoma (LIHC). However, the regulatory mechanisms between acetylation- associated genes (HAGs) and liver cancer stem cells remain unclear. OBJECTIVE: To identify a set of histone acetylation genes (HAGs) with close associations to liver cancer stem cells (LCSCs), and to construct a prognostic model that facilitates more accurate prognosis assessments for LIHC patients. METHODS: LIHC expression data were downloaded from the public databases. Using mRNA expression- based stemness indices (mRNAsi) inferred by One-Class Logistic Regression (OCLR), Differentially Expressed Genes (DEGs) (mRNAsi-High VS. mRNAsi-Low groups) were intersected with DEGs (LIHC VS. normal samples), as well as histone acetylation-associated genes (HAGs), to obtain mRNAsi-HAGs. A risk model was constructed employing the prognostic genes, which were acquired through univariate Cox and Least Shrinkage and Selection Operator (LASSO) regression analyses. Subsequently, independent prognostic factors were identified via univariate and multivariate Cox regression analyses and then a nomogram for prediction of LIHC survival was developed. Additionally, immune infiltration and drug sensitivity analysis were performed to explore the relationships between prognostic genes and immune cells. Finally, the expressions of selected mRNAsi-HAGs were validated in the LIHC tumor sphere by quantitative Reverse Transcription Polymerase Chain Reaction (qRT-PCR) assay and western blot analysis. RESULTS: Among 13 identified mRNAsi-HAGs, 3 prognostic genes (HDAC1, HDAC11, and HAT1) were selected to construct a risk model (mRNAsi-HAGs risk score = 0.02 * HDAC1 + 0.09 * HAT1 + 0.05 * HDAC11). T-stage, mRNAsi, and mRNAsi-HAGs risk scores were identified as independent prognostic factors to construct the nomogram, which was proved to predict the survival probability of LIHC patients effectively. We subsequently observed strongly positive correlations between mRNAsi-HAGs risk score and tumor-infiltrating T cells, B cells and macrophages/monocytes. Moreover, we found 8 drugs (Mitomycin C, IPA 3, FTI 277, Bleomycin, Tipifarnib, GSK 650394, AICAR and EHT 1864) had significant correlations with mRNAsi-HAGs risk scores. The expression of HDAC1 and HDAC11 was higher in CSC-like cells in the tumor sphere. CONCLUSION: This study constructed a mRNAsi and HAGs-related prognostic model, which has implications for potential immunotherapy and drug treatment of LIHC.

Application of noninvasive sampling technique in mitochondrial genome intraspecific phylogeny of the endangered butterfly, Teinopalpus aureus (Lepidoptera: Papilionidae).

The butterfly genus of Teinopalpus, endemic to Asia, embodies a distinct species of mountain-dwelling butterflies with specific habitat requirements. These species are rare in the wild and hold high conservation and research value. Similar to other protected species, the genetic analysis of the rare Teinopalpus aureus poses challenges due to the complexity of sampling. In this study, we successfully extracted DNA and amplified mitochondrial genomic DNA from various noninvasive sources such as larval feces, larval exuviae, larval head capsules, pupal exuviaes, and filamentous gland secretions, all integral parts of butterfly metamorphosis. This was conducted as part of a research initiative focused on the artificial conservation of T. aureus population in Jinggang Shan Nature Reserve. Our findings illustrated the successful extraction of DNA from multiple noninvasive sources, achieved through modified DNA extraction methodologies. Although the DNA concentration obtained from noninvasive samples was lower than that from muscle tissues of newly dead larvae during rearing, all samples met the requirements for PCR amplification and sequencing, yielding complete circular sequences. These sequences are pivotal for both interspecific and intraspecific genetic relationship analysis. Our methods can be extended to other insects, especially scarce species.

Refinement of sensitive azides via in situ generated azole-based metal-organic frameworks towards stable energetic materials.

Energetic materials (EMs) have been widely employed in both military and civilian areas for nearly two centuries. The introduction of high-energy azide anions to assemble energetic metal-organic frameworks (EMOFs) is an efficient strategy to enhance energetic properties. However, azido-based EMOFs always suffer low stabilities to external mechanical stimulation. Herein, we employed an in situ hydrothermal reaction as a technique to refine azide anions with a neutral triazole-cyano-based ligand TrzAt (TrzAt = 2-(1H-1,2,4-triazol-1-yl)acetonitrile) to yield two tetrazole-based EMOFs, namely, [ZnBr(trmetz)]n1 and [Cd(trmetz)2]n2 (Htrmetz = 5-(1,2,4-triazol-1-ylmethyl)-1H-tetrazole). Compound 1 features a closely packed 2D layered network, while compound 2 exhibits a 3D architecture. With azide anions inlaid into a nitrogen-rich and chelating ligand in the EMOFs, compounds 1 and 2 present remarkable decomposition temperatures (Tdec ≥ 300 °C), low impact sensitivities (IS ≥ 32 J) and low friction sensitivities (FS ≥ 324 N). The calculated heat of detonation (ΔHdet) values of 1 and 2 are 3.496 and 4.112 kJ g-1, respectively. In particular, the ΔHdet value of 2 is higher than that of traditional secondary explosives such as 2,4,6-trinitrotoluene (TNT, ΔHdet = 3.720 kJ g-1). These results indicate that EMOFs 1 and 2 may serve as potential replacements for traditional secondary explosives. This work provides a simple and effective strategy to obtain two EMOFs with satisfactory energy densities and reliable stabilities through an in situ hydrothermal technique for desensitization of azide anions.

An injectable nanocomposite alginate-Ca2+ hydrogel for melittin-assisted Ca2+-overload and photothermal cancer therapy.

An injectable nanocomposite alginate-Ca2+ hydrogel embedded with melittin and polyaniline nanofibers was fabricated for Ca2+-overload and photothermal combination cancer therapy. Melittin disrupts the cell membranes and enhances Ca2+ influx significantly, improving Ca2+-overload treatment, while the polyaniline nanofibers endow the hydrogel with glutathione (GSH) depletion and photothermal ability.

Regenerative plantlets with improved agronomic characteristics caused by anther culture of tetraploid potato (Solanum tuberosum L.).

Objective: As the primary means of plant-induced haploid, anther culture is of great significance in quickly obtaining pure lines and significantly shortening the potato breeding cycle. Nevertheless, the methods of anther culture of tetraploid potato were still not well established. Methods: In this study, 16 potato cultivars (lines) were used for anther culture in vitro. The corresponding relation between the different development stages of microspores and the external morphology of buds was investigated. A highly-efficient anther culture system of tetraploid potatoes was established. Results: It was shown in the results that the combined use of 0.5 mg/L 1-Naphthylacetic acid (NAA), 1.0 mg/L 2,4-Dichlorophenoxyacetic acid (2,4-D), and 1.0 mg/L Kinetin (KT) was the ideal choice of hormone pairing for anther callus. Ten of the 16 potato cultivars examined could be induced callus with their respective anthers, and the induction rate ranged from 4.44% to 22.67% using this hormone combination. According to the outcome from the orthogonal design experiments of four kinds of appendages, we found that the medium with sucrose (40 g/L), AgNO3 (30 mg/L), activated carbon (3 g/L), potato extract (200 g/L) had a promotive induction effect on the anther callus. In contrast, adding 1 mg/L Zeatin (ZT) effectively facilitated callus differentiation. Conclusion: Finally, 201 anther culture plantlets were differentiated from 10 potato cultivars. Among these, Qingshu 168 and Ningshu 15 had higher efficiency than anther culture. After identification by flow cytometry and fluorescence in situ hybridization, 10 haploid plantlets (5%), 177 tetraploids (88%), and 14 octoploids (7%) were obtained. Some premium anther-cultured plantlets were further selected by morphological and agronomic comparison. Our findings provide important guidance for potato ploidy breeding.

[Mechanism of active components of "Notoginseng Radix et Rhizoma-Drynariae Rhizoma" in treatment of osteoporosis based on network pharmacology and in vitro cell experiment].

The present study aimed to explore the main active components and potential mechanisms of Panax notoginseng saponins(PNS) and osteopractic total flavone(OTF) in the treatment of osteoporosis(OP) through network pharmacology, molecular docking and in vitro cell experiments, which was expected to provide a theoretical basis for clinical applications. The blood-entering components of PNS and OTF were obtained from literature search and online database, and their potential targets were obtained from Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform(TCMSP) and SwissTargetPrediction. The OP targets were obtained by means of searching Online Mendelian Inheritance in Man(OMIM) and GeneCards. The common targets of the drug and disease were screened by Venn. Cytoscape was used to construct a "drug-component-target-disease" network, and the core components were screened according to the node degree. The protein-protein interaction(PPI) network of the common targets was constructed by STRING and Cytoscape, and the core targets were screened according to the node degree. GO and KEGG enrichment analysis of potential therapeutic targets were carried out by R language. Molecular docking was used to determine the binding activity of some active components to key targets by AutoDock Vina. Finally, HIF-1 signaling pathway was selected for in vitro experimental verification according to the results of KEGG pathway analysis. Network pharmacology showed that there were 45 active components such as leachianone A, kurarinone, 20(R)-protopanaxatriol, 20(S)-protopanaxatriol, and kaempferol, and 103 therapeutic targets such as IL6, AKT1, TNF, VEGFA and MAPK3 involved. PI3K-AKT, HIF-1, TNF and other signaling pathways were enriched. Molecular docking revealed that the core components had good binding ability to the core targets. In vitro experiments found that PNS-OTF could up-regulate the mRNA expression levels of HIF-1α, VEGFA and Runx2, indicating that the mechanism of PNS-OTF in treating OP may be related to the activation of HIF-1 signaling pathway, and thus PNS-OTF played a role in promoting angiogenesis and osteogenic differentiation. In conclusion, this study predicted the core targets and pathways of PNS-OTF in treating OP based on network pharmacology and carried out in vitro experimental verification, which reflected the characteristics of multi-component, multi-target and multi-pathway synergy of PNS-OTF, and provided new ideas for the future clinical treatment of OP.

Epidemiological Characteristics and Spatiotemporal Distribution Patterns of Human Norovirus Outbreaks in China, 2012-2018.

Objective: To clarify the epidemiological characteristics and spatial distribution patterns of human norovirus outbreaks in China, identify high-risk areas, and provide guidance for epidemic prevention and control. Methods: This study analyzed 964 human norovirus outbreaks involving 50,548 cases in 26 provinces reported from 2012 to 2018. Epidemiological analysis and spatiotemporal scanning analysis were conducted to analyze the distribution of norovirus outbreaks in China. Results: The outbreaks showed typical seasonality, with more outbreaks in winter and fewer in summer, and the total number of infected cases increased over time. Schools, especially middle schools and primary schools, are the most common settings of norovirus outbreaks, with the major transmission route being life contact. More outbreaks occurred in southeast coastal areas in China and showed significant spatial aggregation. The highly clustered areas of norovirus outbreaks have expanded northeast over time. Conclusion: By identifying the epidemiological characteristics and high-risk areas of norovirus outbreaks, this study provides important scientific support for the development of preventive and control measures for norovirus outbreaks, which is conducive to the administrative management of high-risk settings and reduction of disease burden in susceptible areas.

Stigmasterol: Remodeling gut microbiota and suppressing tumor growth through Treg and CD8+ T cells in hepatocellular carcinoma.

BACKGROUND: Hepatocellular carcinoma (HCC), the most primary malignant liver tumor and is ranked as the fifth most common malignancy worldwide. Despite various therapeutic approaches being used in clinical practice, the overall effectiveness remains insufficient. Stigmasterol, a compound known for its anti-tumor properties and ability to induce apoptosis in tumor cells, has been found to influenced the composition of the intestinal microbiota. However, the mechanism through which stigmasterol influences the intestinal microbial-host crosstalk in HCC remains elusive. PURPOSE: This study was to investigate whether stigmasterol can remodel gut microbiota, and suppress tumor volume by regulating Treg and IFN-γ+ CD8+ cell in the host with HCC. METHOD: Stigmasterol (at dosages of 0, 50, 100, or 200 mg/kg) was orally administered to Balb/c mice with subcutaneous tumor once every 2 days for 3 weeks. RESULTS: We first found that tumors volume in the group treated with 100 mg/kg stigmasterol were significantly decreased compared with those in the control group (P < 0.05), which exhibited a similar effect as the sorafenib treatment in mice with HCC. This resulted in a significant upregulation of Caspase3, Bax, and P53 expressions, as well as a decrease in Cyclin D1 expression, ultimately leading to a reduction in tumor volume. Additionally, stigmasterol can alter the α and ß diversity of the intestinal flora and significantly increase the abundance of Lactobacillus_johnsonii, Lactobacillus_murinus, and Lactobacillus_reuteri (P<0.05), which can lead to a decrease in the ratio of regulatory T cells (Tregs) to CD8+ T cells in the intestinal tract and tumor tissue, and consequently enhance immune response in the tumor microenvironment (TME) in the host with HCC. CONCLUSION: In this study, we initially utilized different dosages of stigmasterol to intervene in mice with HCC and confirmed its inhibitory effects on tumor growth in vivo, and discovered that stigmasterol affected Lactobacillus johnsonii, Lactobacillus murinus, and Lactobacillus reuteri, resulting in an increased proportion of IFN-γ+ CD8+ T cells and Treg cells in both the intestinal mucosa and tumor tissues, and ultimately leading to increased levels of apoptotic proteins and the subsequent death of tumor cells, which shed light on the effect of stigmasterol on host intestinal tissue and intratumoral immune cells by reshaping the intestinal microbiota, and provide a theoretical foundation for the potential clinical application of stigmasterol in the treatment of HCC.

Exosomal microRNAs in the DLK1-DIO3 imprinted region derived from cancer-associated fibroblasts promote progression of hepatocellular carcinoma by targeting hedgehog interacting protein.

BACKGROUND: Hepatocellular carcinoma (HCC) is the sixth most commonly diagnosed cancer and third leading cause of cancer-related death worldwide in 2020. Exosomes derived from cancer-associated fibroblasts (CAFs-exo) can promote tumor progression in various human cancers. However, the underlying regulatory mechanism controlling how CAFs-exo can promote HCC progression remains poorly understood. METHODS: CAFs and para-cancer fibroblasts (PAFs) were isolated from HCC tissues and corresponding para-cancer tissues, then were cultured in vitro. CAFs and PAFs were characterized by immunofluorescence and western blot (WB) assays. Exosomes were isolated by ultracentrifugation, and characterized by transmission electron microscopy, nanoflow cytometry, and WB assay. The internalization of exosomes by HCC cells was observed under a fluorescence microscope. Cell Counting Kit-8 (CCK-8) assay was used to evaluate cell proliferation. Wound healing and transwell assays were used for migration and invasion experiments. RT-PCR assay was used to examine differentially expressed microRNAs (miRNAs) in exosomes and HCC cells. The TargetScan database was used to predict miRNA target genes. Hedgehog interacting protein (HHIP) expression analysis, prognostic analysis, and enrichment analysis of HHIP-related co-expressed genes were performed using the TIMER, UALCAN, Kaplan-Meier plotter, and LinkedOmics databases. RESULTS: CAFs-exo were internalized by HCC cells. CAFs-exo contributed to the aggressive phenotype of HCC cells, while inhibiting exosome secretion reversed these effects. Mechanistically, miRNAs in the DLK1-DIO3 imprinted region (miR-329-3p, miR-380-3p, miR-410-5p, miR-431-5p) were increased in HCC cells co-cultured with CAFs-exo compared with PAFs-exo. Expression of HHIP, a possible miR-431-5p target gene, was significantly downregulated in HCC cells. Low HHIP expression level in tumor tissues could predict poor prognosis in HCC patients. HHIP-related co-expressed genes were mainly associated with cell adhesion molecules. CONCLUSIONS: CAFs-exo can promote HCC progression by delivering miRNAs in the DLK1-DIO3 imprinted region to HCC cells, subsequently inhibiting HHIP expression. HHIP is a potential prognostic biomarker in HCC.

[Spatiotemporal distribution and multi-source characteristics of microplastics in the soil and water environment of Poyang Lake Wetland, China]. / 鄱阳湖湿地水土环境中微塑料的时空分布及多源性.

The increasing microplastics (MPs) pollution in freshwater wetlands has received global concerns. To investigate the spatiotemporal dynamics of MPs in the wetlands of Poyang Lake, surface water and sediment samples were collected from five rivers entering the lake as well as the confluence of Poyang Lake into the Yangtze River, in both dry and wet seasons. The MPs in water and sediment were extracted by the digestion-filtration method and flotation-separation-digestion-filtration method, respectively. Light microscope, Fourier transform infrared spectroscopy, and scanning electron microscope were used for microplastic characterization. The results showed that the abundance of MPs ranged from 32.1 to 127.3 n·L-1 in water samples, and from 533.3 to 1286.6 n·kg-1 in sediment samples during the wet season. In the dry season, the abundance of MPs ranged from 87.1 to 295.5 n·L-1 in water and from 460.0 to 1368.0 n·kg-1 in sediment. Compared with other freshwater wetlands, Poyang Lake had higher abundance of MPs. There were temporal and spatial differences among regions. The main forms of MPs included beads, fragment, film and fiber, and the corresponding polymer components were mainly polystyrene, polypropy-lene and polyethylene. Beads (35.7% in wet season and 52.0% in dry season) were the main form of MPs in water, while fragment (45.8% in wet season and 69.7% in dry season) was the main form of MPs in sediment. Small size (<0.1 mm) MPs were dominant (>50%) in water and sediment in both seasons. The abundance of MPs with different sizes decreased with the increases of size. The potential main sources of MPs in the wetlands of Poyang Lake included the discharge of industrial wastewater, discharge from urban and rural domestic sewage treatment plants, agricultural and fishing activities, and improper disposal of domestic wastes.

Potassium ferrate followed by alkali-stripping treatment to achieve short-chain fatty acids and nitrogen recovery from waste activated sludge.

Waste activated sludge (WAS) is a potential resource to achieve carbon-neutral goal of wastewater treatment plant. However, the solubilization is always the rate-limiting step for resource recovery from anaerobic digestion of WAS. This study reported a novel strategy, i.e., potassium ferrate (PF) followed by alkali-stripping treatment, to achieve short-chain fatty acids (SCFAs) and nitrogen recovery from WAS. Results showed that whether the stripping process was conducted under alkaline condition or not, the SCFAs production potential was increased rather than reduced. The promoted SCFAs production was due to the accelerated solubilization and hydrolysis stages but the inhibited methanogenesis stage. The SCFAs yield reached 258 mg chemical oxygen demand (COD)/g volatile suspended solids (VSS), and the carbon source, including SCFAs, soluble polysaccharides and proteins, reached 384 mg COD/g VSS. The potentially recovered nitrogen was about 8.71 mg NH4+-N/g VSS. This work may provide some new solutions for enhancing resource recovery from WAS.

[Evaluation of agricultural drought in Luanhe River Basin based on the standardized soil moisture index.] / åŸºäºŽæ ‡å‡†åŒ–åœŸå£¤æ¹¿åº¦æŒ‡æ•°çš„æ»¦æ²³æµåŸŸå†œä¸šå¹²æ—±è¯„ä»·.

Drought is one of the main disasters causing agricultural production losses. Accurate and effective agricultural drought monitoring is of great significance to reduce agricultural production losses. In this study, the standardized soil moisture index for agricultural drought evaluation was established by using the historical time series of soil moisture. Based on the index, the temporal and spatial distribution of drought events in the Luanhe River Basin from 2002 to 2019 were analyzed, and the agricultural drought was quantitatively evaluated. The results showed that soil moisture of most grid units in the Luanhe River Basin obeyed normal distribution during the study period, and that the soil moisture of a few grid units obeyed generalized extreme distribution. The changes of drought and flood in each grid unit had good temporal consistency, that is, the changes of drought and flood was consistent. The changes of drought and flood in the middle of the basin was relatively significant, while the agricultural drought generally showed a trend of reduction. The spatial distribution of agricultural drought severity was not obvious. Relatively speaking, the drought characteristics (the frequency, duration and intensity of drought) were higher in the middle and northwest of the basin and lower in the southeast. The results could provide a reference for agricultural drought prediction and the formulation of drought prevention and mitigation measures in the Luanhe River Basin.

CD155/SRC complex promotes hepatocellular carcinoma progression via inhibiting the p38 MAPK signalling pathway and correlates with poor prognosis.

BACKGROUND: Hepatocellular carcinoma (HCC) is a prevalent malignancy with poor prognosis. As a cell adhesion molecule, poliovirus receptor (PVR/CD155) is abnormally overexpressed in tumour cells, and related to tumour proliferation and invasion. However, the potential role and mechanism of CD155 have not yet been elucidated in HCC. METHODS: Immunohistochemistry, RT-PCR and Western blot assays were used to determine CD155 expression in HCC cell lines and tissues. Cell Counting Kit-8 and colony formation assays were used to examine cell proliferation. Transwell and wound healing assays were used to evaluate cell migration and invasion. Cell apoptosis and cycle distribution were assessed by flow cytometry. Cox regression and Kaplan-Meier analyses were performed to explore the clinical significance of CD155. The role of CD155 in vivo was evaluated by establishing liver orthotropic xenograft mice model. RNA sequencing, bioinformatics analysis and co-immunoprecipitation assay were used to explore the downstream signalling pathway of CD155. RESULTS: CD155 was upregulated in HCC tissues and represented a promising prognostic indicator for HCC patients (n = 189) undergoing curative resection. High CD155 expression enhanced cell proliferation, migration and invasion, and contributed to cell survival in HCC. CD155 overexpression also induced epithelial-mesenchymal transition in HCC cells. CD155 function in HCC involved SRC/p38 MAPK signalling pathway. CD155 interacted with SRC homology-2 domain of SRC and promoted SRC activation, further inhibiting the downstream p38 MAPK signalling pathway in HCC. CONCLUSIONS: CD155 promotes HCC progression via the SRC/p38 MAPK signalling pathway. CD155 may represent a predictor for poor postsurgery prognosis in HCC patients.

Comprehensive Analysis of HHLA2 as a Prognostic Biomarker and Its Association With Immune Infiltrates in Hepatocellular Carcinoma.

Background: Inhibitory immune checkpoint proteins promote tumor immune escape and are associated with inferior patient outcome. However, the biological functions and regulatory roles of one of its members, HHLA2, in the tumor immune microenvironment have not been explored. Methods: RandomForest analyses (371 cases), qRT-PCR (15 cases), and immunohistochemical staining (189 cases) were used to validate the prognostic value of HHLA2 in hepatocellular carcinoma (HCC) patients. Bioinformatic analyses were further performed to explore the biological functions and potential signaling pathways affected by HHLA2. Moreover, ESTIMATE, single sample gene set enrichment analysis, CIBERSORT, TIMER, and other deconvolution methods were used to analyze the composition and infiltration level of immune cells. Multiplex immunofluorescence assays were employed to validate the fractions of suppressive immune cells, and HHLA2-related molecular alterations were investigated. Finally, the clinical response to chemotherapy and immune checkpoint blockade was predicted by TIDE, Submap, and several other in silico analyses. Results: RandomForest analysis revealed that HHLA2 was the most important inhibitory immune checkpoint associated with HCC patient prognosis (relative importance = 1). Our HCC cohorts further revealed that high HHLA2 expression was an independent prognostic biomarker of shorter overall survival (P<0.01) and time to recurrence (P<0.001) for HCC patients. Bioinformatics experiments revealed that HHLA2 may accelerate the cell cycle of cancer cells. Additionally, we found that high expression of HHLA2 was associated with immune infiltrates, including some immunosuppressive cells, cytokines, chemokines, and corresponding receptors, resulting in an immunosuppressive environment. Notably, HHLA2 expression was positively correlated with the infiltration of exhausted CD8+ T cells, which was validated by immunofluorescence. Genomic alteration analyses revealed that promoter hypermethylation of HHLA2 may be associated with its low expression. More importantly, patients with high HHLA2 expression may be more sensitive to chemotherapy and have better responses to immunotherapy. Conclusions: High expression of HHLA2 is an independent prognostic biomarker for HCC patients. It can activate the cell cycle and foster an immunosuppressive tumor microenvironment by enriching exhausted CD8+ T cells. Promoter hypermethylation might lead to low expression of HHLA2 in HCC. Thus, targeting HHLA2 may be a practical therapeutic strategy for HCC patients in the future.

Hsa_circ_0003945 promotes progression of hepatocellular carcinoma by mediating miR-34c-5p/LGR4/ß-catenin axis activity.

Accumulating evidence suggests that circular RNAs (circRNAs) play essential roles in regulating cancer progression, but many circRNAs in hepatocellular carcinoma (HCC) remain unknown. Dysregulated circRNAs in HCC were identified through bioinformatics analysis of Gene Expression Omnibus data sets. Quantitative real-time PCR (qRT-PCR), Sanger sequencing, RNase R digestion and actinomycin D treatment were conducted to confirm the characterization of circRNAs. CCK-8, wound-healing and Transwell assays were performed to assess the functional roles of Hsa_circ_0003945 (Circ_0003945) in HCC cell lines. Subcellular fractionation and fluorescence in situ hybridization (FISH) were performed to locate Circ_0003945 in HCC cells. Dual-luciferase reporter assay was executed to verify the binding of Circ_0003945 to microRNAs (miRNAs) or the miRNAs to their target genes. In this study, we found that Circ_0003945 was upregulated in HCC tissue, and higher Circ_0003945 expression was positively correlated with tumour size and tumour stage. Furthermore, high plasma levels of circulating Circ_0003945 were confirmed in HCC patients compared with those in non-HCC groups. The functional experiments revealed that overexpression or knockdown of Circ_0003945 promoted or attenuated tumour growth and migration, respectively. Mechanistically, Circ_0003945 might exert as a miR-34c-5p sponge to upregulate the expression of leucine-rich repeat-containing G protein-coupled receptor 4 (LGR4), activating the ß-catenin pathway, and finally facilitating HCC progression. Additionally, a ß-catenin activator could reverse the effect of Circ_0003945 knockdown. In conclusion, Circ_0003945 exerts a tumour-promoting role in HCC cells by regulating the miR-34c-5p/LGR4/ß-catenin axis, which may be a potential target for HCC therapy.

High serum soluble CD155 level predicts poor prognosis and correlates with an immunosuppressive tumor microenvironment in hepatocellular carcinoma.

BACKGROUND: Hepatocellular carcinoma (HCC) is one of the most prevalent malignancies with poor prognosis. There is no research about the clinical significance of serum soluble CD155 (sCD155) level for HCC. We aim to explore the prognostic and diagnostic value of sCD155 in HCC patients undergoing curative resection. METHODS: Serum sCD155 level in HCC patients was determined by enzyme-linked immunosorbent assay. The prognostic significance of sCD155 was evaluated by Cox regression and Kaplan-Meier analyses. CD155 expression and biomarkers of immune cells in HCC tissues were detected by immunohistochemistry staining. The diagnostic significance of sCD155 was evaluated using receiver operating characteristic curve. RESULTS: Serum sCD155 level was significantly increased in HCC patients and predicted poor prognosis. The prognostic value of sCD155 remained in low recurrent risk subgroups of HCC. Serum sCD155 level was positively related to CD155 expression in HCC tissues. High serum sCD155 level was associated with decreased numbers of CD8+ T cells and CD56+ NK cells and increased number of CD163+ M2 macrophages. Serum sCD155 level had better performance in distinguishing HCC patients from healthy donors and patients with chronic liver conditions than α-fetoprotein. Among patients with α-fetoprotein ≤ 20 ng/ml, serum sCD155 level could differentiate HCC patients from non-HCC patients. CONCLUSION: Serum sCD155 level represents a promising biomarker for diagnosis and prognosis of HCC. High serum sCD155 level may reflect an immunosuppressive tumor microenvironment in HCC.

Role and significance of water and acid washing on biochar for regulating methane production from waste activated sludge.

Methane recovered from anaerobic digestion of waste activated sludge (WAS) can be used as the energy supplement of the wastewater treatment plant, benefiting to its carbon-neutral operation. In order to enhance methane production, biochar (BC) has been widely selected as conductive material to build direct interspecies electron transfer (DIET) in anaerobic digestion of WAS. However, the role and significance of washing strategies, including water and acid washing, on BCs for regulating methane production have not been reported. This study selected the frequently used woody- (W) and straw (S)-BCs as mode. Compared to raw W-BC, water and acid washing W-BC increased the methane yields by 19.1% and 15.7%, respectively. Differently, the methane yields among raw, water and acid washing S-BCs were similar. Mechanism study showed that both the two washing strategies optimized the properties of raw W-BC for promoting methane production. Water and acid washing W-BCs increased the electron transfer functional groups, such as ketones and quinones, which were not observed in S-BCs. Moreover, the electron-active microorganisms were enriched with the presence of water and acid washing W-BCs, and the predominant pathway for methane production shifted from hydrogentrophic to acetotrophic and DIET methanogenesis, while the microbial communities, including bacteria and archaea, were similar with the presence of raw, water and acid washing S-BCs. These findings of this work provide some new insights for production improvement regulation of methane from anaerobic digestion of wastes induced by BCs.

Freezing-low temperature treatment facilitates short-chain fatty acids production from waste activated sludge with short-term fermentation.

This study proposed a novel and high-efficiency strategy, i.e., freezing followed by low-temperature thermal treatment, to significantly promote short-chain fatty acids (SCFAs) production from waste activated sludge compared to traditional freezing/thawing treatment. The maximal production of SCFAs was 212 mg COD/g VSS with a shortened retention time of five days, and the potentially recovered carbon source, including SCFAs, soluble polysaccharides and proteins, reached 321 mg COD/g VSS, increased by 92.1 and 28.3% compared to sole freezing and thermal treatment. Both the solubilization and hydrolysis steps of WAS were accelerated, and the acid-producing microorganisms, such as Macellibacteroides, Romboutsia and Paraclostridium, were greatly enriched, with a total abundance of 13.9%, which was only 0.54% in control. Interestingly, the methane production was inhibited at a shortened retention time, resulting in SCFAs accumulation, whereas it was increased by 32.0% at a longer sludge retention time, providing a potential solution for energy recovery from WAS.

Stepwise freezing-thawing treatment promotes short-chain fatty acids production from waste activated sludge.

Waste activated sludge (WAS), as the byproducts of wastewater treatment plants, has been greatly produced. With high cost and environmental risk of WAS disposal, to explore a low-cost and environment-friendly technology has been a great challenge. Considering that WAS is a collection of organic matters, anaerobic fermentation has been selected as a sustainable way to simultaneously recover resources and reduce environmental pollution. To recover short-chain fatty acids (SCFAs) has gained great concern because of the high value-added application and high-efficiency production process. Considering the temperature in some areas of the world can reach to below 0 °C, this study proposed an efficient strategy, i.e., stepwise freezing and thawing treatment, to promote SCFAs production. The maximal production of SCFAs, i.e., 246 mg COD/g volatile suspended solid, was obtained with the shortened retention time of five days. Mechanistic studies showed that the solubilization of both extracellular polymeric substances (EPSs) and microbial cells could be accelerated, with the EPSs removal of 58.3% for proteins and 59.0% for polysaccharides. Also, the hydrolysis process was promoted to provide more substrates for subsequent acidogenisis, and the functional microorganisms, such as Romboutsia, Paraclostridium, Macellibacteroides and Conexibacter, were greatly enriched, with a total abundance of 26.2%. Moreover, compared to control, methanogenesis was inhibited at a shortened sludge retention time (e.g., five days), which benefited to the accumulation of SCFAs, but the methane production was increased by 25.2% at a longer sludge retention time (e.g., ten days). Thus, these findings of this work may provide some new solutions for the enhanced resource recovery from WAS, and further for carbon-neutral operation of wastewater treatment plants.