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Water-saving irrigation technology has been studied for many years, but due to insufficient investment in research and development of irrigation equipment, it cannot be effectively promoted and applied. In order to improve the water use efficiency of farmland irrigation, the automatic monitoring of groundwater level change, soil moisture change, and crop seasonal growth status is realized in the ditch or pump well irrigation area. An intelligent measuring terminal for water-saving irrigation based on RN2026 microcontroller is developed. The terminal uses the coefficient of “water conversion by electricity” to automatically convert the water consumption of farmers and uses the price mechanism of water rights to affect farmers’ water conservation. In combination with changes in groundwater level, crop evapotranspiration, and dynamic changes in soil moisture, precision irrigation strategies are automatically generated to guide farmers to use water rationally. Through the function and performance experiments, the results show that the error of the coefficient of “water conversion by electricity” is less than 1%, the error of electric energy measurement is less than 0.5%, the estimated value of average water use right per unit arable area is consistent with the actual value, and the predicted value of daily average soil moisture change is consistent with the measured value. The equipment has strong adaptability to the environment, high measurement accuracy, strong data processing ability, low cost, and other characteristics. It meets the requirements of water-saving irrigation technology in practical applications and has good promotion and application value.

Carnosic acid protects against doxorubicin-induced cardiotoxicity by inhibiting NLRP3 inflammasome activity, which is related to the upregulation of the Nrf2/HO-1 transcription pathway.

We propose an adaptive planewave method for eigenvalue problems in electronic structure calculations. The method combines a priori convergence rates and accurate a posteriori error estimates into an effective way of updating the energy cut-off for planewave discretizations, for both linear and nonlinear eigenvalue problems. The method is error controllable for linear eigenvalue problems in the sense that for a given required accuracy, an energy cut-off for which the solution matches the target accuracy can be reached efficiently. Further, the method is particularly promising for nonlinear eigenvalue problems in electronic structure calculations as it shall reduce the cost of early iterations in self-consistent algorithms. We present some numerical experiments for both linear and nonlinear eigenvalue problems. In particular, we provide electronic structure calculations for some insulator and metallic systems simulated with Kohn--Sham density functional theory (DFT) and the projector augmented wave (PAW) method, illustrating the efficiency and potential of the algorithm.

In order to get the law of rainwater infiltration and the law of progressive water damage, the slope of Tianshang bridge foundation in Yunnan Province is taken as the engineering basis. The site investigation of the damaged slope is carried out, The traditional Green-Ampt model is improved by considering the ponding effect of dynamic water flow on the slope surface. Based on the variation characteristics of wetting front obtained by the improved infiltration model, the progressive failure process of slope with continuous infiltration is simulated by FLAC3D software. The results show that: the water damage of Tianshang bridge slope is mainly caused by the slope absorbing a lot of rainwater in a short time, which has obvious time discontinuity characteristics in time, and shows obvious multi-layer progressive failure characteristics in failure space. The wetting front characteristics obtained by the improved Green-Ampt infiltration model are more consistent with the engineering practice, the water accumulation effect of surface water gradually weakens with the increase of the wet front depth. the slope failure develops gradually from the toe to the top and from shallow to deep, and the intermittent failure characteristics provide surplus time for the engineering treatment, the gravel soil slope should be treated after a small-scale water damage occurs.

ObjectiveSolid tumours respond poorly to immune checkpoint inhibitor (ICI) therapies. One major therapeutic obstacle is the immunosuppressive tumour microenvironment (TME). Cancer-associated fibroblasts (CAFs) are a key component of the TME and negatively regulate antitumour T-cell response. Here, we aimed to uncover the mechanism underlying CAFs-mediated tumour immune evasion and to develop novel therapeutic strategies targeting CAFs for enhancing ICI efficacy in oesophageal squamous cell carcinoma (OSCC) and colorectal cancer (CRC).DesignAnti-WNT2 monoclonal antibody (mAb) was used to treat immunocompetent C57BL/6 mice bearing subcutaneously grafted mEC25 or CMT93 alone or combined with anti-programmed cell death protein 1 (PD-1), and the antitumour efficiency and immune response were assessed. CAFs-induced suppression of dendritic cell (DC)-differentiation and DC-mediated antitumour immunity were analysed by interfering with CAFs-derived WNT2, either by anti-WNT2 mAb or with short hairpin RNA-mediated knockdown. The molecular mechanism underlying CAFs-induced DC suppression was further explored by RNA-sequencing and western blot analyses.ResultsA negative correlation between WNT2+ CAFs and active CD8+ T cells was detected in primary OSCC tumours. Anti-WNT2 mAb significantly restored antitumour T-cell responses within tumours and enhanced the efficacy of anti-PD-1 by increasing active DC in both mouse OSCC and CRC syngeneic tumour models. Directly interfering with CAFs-derived WNT2 restored DC differentiation and DC-mediated antitumour T-cell responses. Mechanistic analyses further demonstrated that CAFs-secreted WNT2 suppresses the DC-mediated antitumour T-cell response via the SOCS3/p-JAK2/p-STAT3 signalling cascades.ConclusionsCAFs could suppress antitumour immunity through WNT2 secretion. Targeting WNT2 might enhance the ICI efficacy and represent a new anticancer immunotherapy.

The abnormality of ribosomal biogenesis is found to be involved in carcinogenesis and progression in various types of cancer. RPL22L1 gene is a highly homologous paralog of RPL22, both of which belong to the L22E family of ribosomal proteins and take part in encoding a cytoplasmic ribosomal protein that is a component of the 60S subunit of ribosomes. Our previous work has established a CRISPR/Cas9 genome-wide knockout screening profile and developed a two-component mixture model to identify cancer-specific essential genes in 436 cancer cell lines. Bioinformatics analysis reveals that RPL22L1 acts as a feature gene to predict cancer-specific essential genes in human cancer. Previous studies suggest that RPL22L1 is correlated with tumor progression and adverse prognosis in several cancer types, while its role in hepatocellular carcinoma (HCC) is unknown. In the present study, we found that RPL22L1 is highly expressed in embryonic stem cells (ES) and liver progenitor cells, which were acquired from a vitro hepatic differentiation model originating from ES cells. Hence, we hypothesized that RPL22L1 enhances cancer stem cell properties in HCC. Functional assays showed that RPL22L1 would promote sphere formation in vitro. qRT-PCR and Western Blotting results indicated that RPL22L1 could promote stemness-related gene expressions such as CD133, CD44, and SOX9 in HCC cell lines. Overexpression of RPL22L1 increased aldehyde dehydrogenase (ALDH) activity, which was decreased when RPL22L1 was silenced with short hairpin RNA (shRNA). Extreme limiting dilution assay showed that RPL22L1 overexpression stimulated self-renewal ability in immunodeficient mice. Furthermore, higher RPL22L1 expression can also promote cell proliferation and epithelial-mesenchymal-transition, as well as drug resistance to 5-Fluorouracil (5-FU) and cisplatin (CDDP). Silver nitrate staining demonstrated that RPL22L1 overexpression correlated with a larger size and number of nucleoli. The expression of RPL22L1 was also associated with adverse prognosis in HCC clinical samples. In conclusion, our study reveals aberrant ribosomal protein activity in HCC and the role of RPL22L1 in promoting HCC tumor progression. Citation Format: JinLin Huang, Zhenyang Guo, Jie Luo, Beilei Liu, Lanqi Gong, Xiaona Fang, Dora Lai-wan Kwong, Xinyuan Guan. Overexpression of ribosomal L22-like1 (RPL22L1) enhances cancer stem cell properties and promotes tumor progression in hepatocellular carcinoma (HCC) [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 1489.

Esophageal squamous carcinoma (ESCC) is one of the leading malignancies worldwide, especially in Asian countries, largely threatening human life. The 5-year survival rate of ESCC is less than 20%, and patients with distant metastases have dismal survival with Citation Format: Hongyu Zhou, Beilei Liu, Ching Ngar Wong, Xinyuan Guan. GPRC5A drives an implantation-like phenotype in early-stage lung disseminated cancer cells of esophageal squamous carcinoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 1293.

Hepatocellular carcinoma (HCC) ranks as the fifth most common cancer and the second leading cause of cancer-related mortality worldwide. The overall prognosis of HCC patients is very poor because it is susceptible to recurrence and metastasis. According to the cancer stem cell (CSC) model, there is a small subset of cancer cells within the tumor bulk that is responsible for tumor relapse and metastasis. CSCs represent cells that can self-renew, initiate tumors, and keep resistant to anti-cancer treatment. However, the fate and signatures of CSCs in HCC remain elusive. Our previous in vitro hepatocyte differentiation model successfully mimicked the liver developmental process from the stage of embryonic stem cells, endoderm, liver progenitor cells, and premature hepatocytes, together with one pair of nontumor and tumor tissue from an HCC patient. We found that the expression of keratin 17 (KRT17) reached its peak at the liver progenitor stage and was down-regulated along with liver development, which shared the same expression pattern with keratin 19 (CK19), a well-known HCC stemness-associated gene. Single-cell RNA sequencing in 9 HCC clinical samples from the GEO dataset revealed that KRT17 was expressed in less than 1% of tumor cells. Moreover, KRT17 shared similar expression patterns among hepatocytes, non-stem like HCC and stem like HCC subpopulations with a panel of hepatic stem cell markers and showed reverse trends with a list of hepatic markers. Clinically, KRT17 was highly expressed in cancer tissues compared to normal counterparts and positively correlated with poor overall survival. Functional assays indicated that KRT17 could considerably facilitate the self-renewal, growth, and metastasis properties of HCC both in vitro and in vivo. Also, KRT17 endowed HCC cells with enhanced resistance to sorafenib treatment. Moreover, the expressions of the CSC marker (CD133) and oncofetal drivers (NANOG and KLF4) were significantly upregulated upon KRT17 overexpression. Bioinformatic analysis using gene expression profiles of patients from the TCGA database revealed that development- and differentiation-related biological processes were dramatically enriched in KRT17-high patients. The underlying molecular mechanism of how KRT17 augments HCC stemness still needs further study. Overall, our study uncovered a new connection between KRT17 and HCC stemness and progression. KRT17 might be a novel marker of CSC and a prognostic biomarker for the treatment of HCC patients. Citation Format: Xiaona Fang, Shan Liu, Liuxian Ban, Jiao Huang, Jie Luo, Lanqi Gong, Baifeng Zhang, Beilei Liu, Jinlin Huang, Yuma Yang, Ching Ngar Wong, Qian Yan, Xin-Yuan Guan. Identification and characterization of the role of KRT17 in the stemness regulation of hepatocellular carcinoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 2441.

PPM1G is a member of protein phosphatase family, marked as Mg2+/Mn2+ dependent 1G. PPM1G protein mainly has three domains: phosphatase domains at both ends and acidic domain (AD) in the middle. It was highly expressed in both embryonic stem cells and hepatocellular carcinoma (HCC) by mRNA-seq data. From pan-cancer view, it was up-regulated in tumor tissue of 24 pan-cancer types and predicted worse overall survival (OS) for 9 cancers. In GSE40267 dataset, PPM1G overexpressed significantly in lung metastasis HCC patients than patients without metastasis. And both from TCGA and ICGC-JP database analysis showed PPM1G was an independent poor prognostic factor for both OS and disease-free survival (DFS) in HCC patients. In vitro, our functional experiments showed that the stemness features such as sphere formation, ALDH assay and atRA (Tretinoin) assay were undermined by knock down (KD) of PPMIG. In HepG2 and 8024 KD cell lines, the tumor capacity of proliferation, cell-cycle, foci-formation, migration invasion and cisplatin induced apoptosis were also impaired. In vivo, KD of PPM1G strongly suppressed the subcutaneous and liver orthotopic tumor formation and lung metastasis and cancer stemness by limited dilution experiment was also significantly weakened. For the mechanism exploration, firstly, GO analysis showed genes highly positive correlated with PPM1G (Spearman R > 0.5) enriched in mRNA splicing. Secondly, YBX1 (role as RNA m5C and m6A reader protein and chemotherapy resistance gene) was screened out by three database (RBP2GO, HiPredict and GEPIA) followed with in vivo CO-IP experiment and immunofluorescence colocalization. The YBX1 expression level was not affected by PPM1G from Western Blot. Lastly, the AD of PPM1G but not the phosphatase domain was interacted with YBX1 by PPM1G cut-off and CO-IP experiments. So, the above evidence proved that PPM1G could interact with YBX1 to promote HCC stemness and tumor growth and metastasis. Citation Format: Shan Liu, Xiaona Fang, Jie Luo, Jiao Huang, Yuma Yang, Beilei Liu, Jinlin Huang, Ching Ngar Wong, Baifeng Zhang, Lanqi Gong, Liuxian Ban, Qian Yan, Xinyuan Guan. PPM1G enhances hepatocellular carcinoma stemness via interacting with YBX1 [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 2445.

Background: Nasopharyngeal carcinoma (NPC) is a highly undifferentiated squamous malignancy with dense infiltration of dysfunctional T cells that is endemic in East and Southeast Asia. Based on multicentral single-cell RNA sequencing (scRNA-seq) analysis, we have identified SRY-Box Transcription Factor 4 (SOX4), an essential developmental transcription factor, is specifically and highly expressed by NPC cells and contributes to tumor growth and distant metastasis. SOX4 upregulation also promotes ZIP14-mediated intracellular zinc uptake, leading to competitive deprivation of free zinc in the tumor microenvironment (TME), which inhibits zinc-dependent T cell receptor (TCR) activation. Thus, our study provides experimental insights into how NPC develops and evades immune surveillance. Methods: scRNA-seq data and bulk RNA-Seq data from our lab and GEO database were integrated and analyzed on a bioinformatic basis. Immunohistochemistry staining was performed to validate the SOX4 protein level in an independent clinical cohort. Cell lines, patient-derived xenografts, and humanized mouse models were generated to evaluate the in vitro and in vivo function of SOX4. Flow cytometry was used for T-cell immunophenotyping. Immunoprecipitation was used to study protein-DNA interaction. The dual-luciferase reporter assay was utilized to demonstrate the transcriptional regulation of SOX4 on the ZIP14 promoter. qRT-PCR, immunoblotting, and immunofluorescence staining were also employed for functional studies. Result: Our transcriptome analysis showed SOX4 was frequently overexpressed by NPC cells, compared to infiltrating immune and stromal cells. Increasing SOX4 expression in normal nasopharyngeal epithelial cells enhanced proliferation, stemness, migration, and invasion. SOX4 also transcriptionally upregulated ZIP14 in NPC cells which promoted the uptake of free zinc from the TME. Thus, competitive deprivation of zinc between NPC cells and T cells in the TME inhibited TCR signaling and T-cell cytotoxicity, leading to NPC immune evasion. Conclusion: This study reveals the SOX4-ZIP14-Zinc axis as a vital mechanism for simultaneously inducing tumor development and immunosuppression in the NPC microenvironment. Therapeutic inhibition of SOX4 or ZIP14 is a new strategy for NPC patients and potentially synergizes with PD-1/PD-L1 blockades to overcome immunosuppressive signals in the TME. Citation Format: Yuma Yang, Lanqi Gong, Jie Luo, Xiaona Fang, Jiao Huang, Lu Bai, Qin Liu, Beilei Liu, Shan Liu, Jinlin Huang, Ching Ngar Wong, Baifeng Zhang, Danyang Zheng, Yu Zhang, Wei Dai, Xinyuan Guan. SOX4 promotes tumor development and immune evasion via disruption of zinc homeostasis between nasopharyngeal carcinoma cells and T cells. [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 4450.

Immune evasion is one of the important factors leading to the progression of hepatocellular carcinoma(HCC). Restricted antigen presentation by decrease of MHC-I remains a huge barrier to triggering an immune attack. However, knowledge is limited on how to enhance antigen presentation of cancer cells. Here we identify that FASN inhibition by shRNA and inhibitors could boost MHC-I level on HCC cell surface. FASN deficiency also promotes antigen presentation and augments CD8+ T cell cytotoxicity. Mechanistically, FASN knock-down decreases the degradation of MHC-I in lysosomes. Moreover, ZDHHC3-mediated palmitoylation participates in this process. In vivo, orthotopic HCC mouse model shows that Fasn knock-down greatly improves the cytotoxicity of tumor-infiltrating CD8+ T cells and suppresses HCC tumor growth, indicating that inhibiting FASN is a promising way to enhance the efficiency of immunotherapy in HCC. Citation Format: Jiao Huang, Wai Ying Tsang, Xiaona Fang, Yu Zhang, Jie Luo, Lanqi Gong, Baifeng Zhang, Ching Ngar Wong, Beilei Liu, Jinlin Huang, Yuma Yang, Shan Liu, Zhihong Li, Xinyuan Guan. FASN inhibition overcomes immune evasion by upregulating MHC-I in hepatocellular carcinoma. [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 4452.

Poorly differentiated tumors usually exhibit phenotypes similar to that of their developmental precursor cells. Tumor cells that acquire the lineage progenitor cells feature usually exploit developmental signaling to potentiate cancer progression. However, the underlying molecular events remain elusive. In this study, based on analysis of an in vitro hepatocyte differentiation model, the maternal factor PGC7 (also known as DPPA3, STELLA) was found closely associated with liver development and tumor differentiation in hepatocellular carcinoma (HCC). Expression of PGC7 decreased during hepatocyte maturation and increased progressively from well-differentiated HCCs to poorly differentiated HCCs. Whole-genome methylation sequencing found that PGC7 could induce promoter demethylation of genes related to development. Pathway-based network analysis indicated that downstream targets of PGC7 might form networks associated with developmental transcription factor activation. Overexpression of PGC7 conferred progenitor-like features of HCC cells both in vitro and in vivo. Mechanism studies revealed that PGC7 could impede nuclear translocation of UHRF1, and thus facilitate promoter demethylation of GLI1 and MYCN, both of which are important regulators of HCC self-renewal and differentiation. Depletion or inhibition of GLI1 effectively downregulated MYCN, abolished the effect of PGC7, and sensitized HCC cells to sorafenib treatment. In addition, we found a significant correlation of PGC7 with GLI1/MYCN and lineage differentiation markers in clinical HCC patients. PGC7 expression might drive HCC toward a “dedifferentiated” progenitor lineage through facilitating promoter demethylation of key developmental transcription factors; further inhibition of PGC7/GLI1/MYCN might reverse poorly differentiated HCCs and provide novel therapeutic strategies.

Chronic hepatitis B virus (HBV) infection is strongly associated with the initiation and development of hepatocellular carcinoma (HCC). However, the genetic alterations and pathogenesis mechanisms remain significantly unexplored, especially for HBV-induced metabolic reprogramming. Analysis of integration breakpoints in HBV-positive HCC samples revealed the preferential clustering pattern within the 3′-end of X gene in the HBV genome, leading to the production of C-terminal truncated X protein (Ct-HBx). In this study, we not only characterized the oncogenic role of two Ct-HBx (HBx-120 and HBx-134) via in vitro and in vivo functional assays but also deciphered their underlying molecular mechanisms. Gene expression profiling by transcriptome sequencing identified potential targets of Ct-HBx and novel malignant hallmarks such as glycolysis, cell cycle, and m-TORC1 signaling in Ct-HBx-expressing cells. TXNIP, a well-established regulator of glucose metabolism, was shown to be downregulated by Ct-HBx and play a pivotal role in Ct-HBx-mediated HCC progression. Suppression of TXNIP is frequently observed in HCC patients with Ct-HBx expression and significantly (P = 0.015) correlated to a poorer prognosis. Re-introduction of TXNIP attenuated the metabolic reprogramming induced by the Ct-HBx and inhibited the tumor growth in the mice model. Further study suggested that Ct-HBx could downregulate TXNIP via a transcriptional repressor nuclear factor of activated T cells 2 (NFACT2). Collectively, our findings indicate that TXNIP plays a critical role in Ct-HBx-mediated hepatocarcinogenesis, serving as a novel therapeutic strategy in HCC treatment.

This article discusses the impact of corporate managers' green environmental awareness and strategic intelligence on the formulation of corporate green product innovation strategies and their relationship with marketing performance. As the pre-factors affecting innovation, the green environmental awareness and strategic intelligence will affect the formulation and implementation of corporate green product innovation strategies, thereby promoting corporate performance. According to the regression analysis of 367 questionnaires from heavily polluting enterprises, it can be seen that managers' green environmental awareness is positively correlated with green product technological innovation and design innovation strategies. Managers' meta-cognitive strategic intelligence does not have a moderating effect on the relationship between green environmental awareness and green product technological innovation strategy, but has a significant positive moderating effect on the green product design innovation strategy. Managers' motivational strategic intelligence has a positive influence on the relationship between green environmental awareness and green product technology and design innovation strategy. In addition, the implementation of green product innovation strategies directly promotes the improvement of marketing performance. The above conclusions can provide inspiration or companies during the new opportunity period to transform managers' environmental awareness and strategic intelligence into corporate performance by using green product innovation strategies.

Compositions of resin composite exhibit cytotoxicity, especially Triethylene-glycol-dimethacrylate (TEGDMA), yet the underlying mechanisms and its relationship with filler content are poorly understood. Here, specimens of five composites (VITA LC, VITA ZETA, Z350, Filtek P60, and AP-X), containing different filler size and weight, were immersed into culture medium for 72 h. After TEGDMA quantification, the resin composite eluates were used to incubate HGFs. Cellular viability was evaluated. Total reactive oxygen species (ROS) and mitochondrial ROS were detected to assess oxidative stress. Adenosine triphosphate and cytochrome c oxidase (CcO) activity, mitochondrial membrane potential and morphology, mitochondrial biogenesis regulators were analyzed to evaluate mitochondrial functions. Results showed that TEGDMA release negatively correlated to filler size and weight of tested composites. Although cell viability reduction was not significant, total and mitochondrial ROS production showed a positive relationship with the amount of TEGDMA in composite eluates. Furthermore, the expression of mitochondrial biogenesis markers and mitochondrial fusion protein, were markedly elevated in TEGDMA rich eluates, especially in VITA-LC group, shown as elongated mitochondrial morphology and aberrant mitochondrial functions. Overall, TEGDMA could elute easier from those resin composites with less filler content and cause oxidative stress in HGFs via mitochondria dysregulation. These data can be instructive to optimize the synthesis of resin composites from the perspective of biocompatibility. © 2019 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 107A: 1132-1142, 2019.

Background MicroRNAs (miRNAs) have been shown to commonly contribute to cardiac hypertrophy (CH). The aim of this study was to test the hypothesis that miR‐200c plays an important role in the progression of CH by targeting myosin light chain kinase (MLCK/MYLK). Methods and results Cardiac hypertrophy was induced by aortic banding (AB) in rats. Cellular hypertrophy in neonatal rat cardiomyocytes (NCMs) was induced by AngII treatment. Echocardiography, histology and molecular measurements were used to assess the results of the experiments. The levels of apoptosis and reactive oxygen species (ROS) were also measured. Quantitative real‐time PCR (qRT‐PCR) and Western blotting were used to measure mRNA and protein levels respectively. The present results showed that miR‐200c expression was increased in response to CH both in vivo and in vitro. The down‐regulation of miRNA‐200c by a specific inhibitor markedly ameliorated CH resulting from AngII treatment, and the mRNA levels of atrial natriuretic peptide, brain natriuretic peptide and β‐myosin heavy chain were simultaneously decreased. Notably, minimal apoptosis and ROS accumulation were identified in AngII‐induced hypertrophic cardiomyocytes. Conversely, the up‐regulation of miR‐200c using specific mimics reversed these effects. Mechanistic investigations demonstrated that the MLCK gene is a direct target of miR‐200c; an increase in miR‐200c levels led to a decrease in the expression of MLCK and its downstream effector, p‐MLC2, while miR‐200c inhibition increased the expression of these proteins. Furthermore, inhibiting MLCK impaired the anti‐hypertrophic effects contributions produced by the knockdown of miR‐200c. Conclusion Our studies suggest that miR‐200c may serve as a potential therapeutic target that could delay hypertrophy. We have also uncovered a relationship between miR‐200c and MLCK, identifying MLCK as a direct mediator of miR‐200c.

Hepatocellular carcinoma (HCC) is one of the frequently diagnosed cancers worldwide and has emerged as the main cause of cancer-related death with a very poor prognosis, which is closely related to cancer stem cell (CSC), a subset of cells known to be a root of tumor recurrence, metastasis, and chemoresistance in HCC. However, the characteristics of CSCs in hepatocellular carcinoma (HCC) are still elusive. Our previous in vitro hepatocyte differentiation model revealed that the expression of anti-silencing function 1B histone chaperone (ASF1B) was remarkably upregulated in the stem cell-like stages. In HCC patients, ASF1B was highly expressed in cancer tissues compared with normal counterparts and was positively correlated with poor survival. Functional assays indicated that ASF1B could considerably promote the self-renewal, growth, and metastasis of HCC both in vitro and in vivo. Mechanistically, ASF1B could directly bind to histone H3 Variant H3.3 and enhance its incorporation into the promoter regions of certain stemness- and metastasis-associated genes including SOX9, KLF4, and TBX3, contributing to their augmented transcriptional and protein expression levels. H3.3 knock-down decreased the expression of these stemness and metastasis-associated genes, eliminating the stemness and oncogenic properties of ASF1B. Overall, our study revealed that ASF1B could facilitate HCC stemness and progression via H3.3-dependent transcriptional reprogramming. ASF1B might be a novel diagnostic and prognostic biomarker for HCC patients. Citation Format: Xiaona Fang, Beilei Liu, Shan Liu, Yu Zhang, Lanqi Gong, Qian Yan, Xinyuan Guan. ASF1B potentiates stem cell traits and tumor progression in hepatocellular carcinoma via histone H3.3-dependent transcriptional reprogramming [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 6065.

Background: Hepatocellular carcinoma (HCC) is one of the most common and most difficult to cure malignancies worldwide. Identifying new effective therapeutic targets is of great significance for the management of HCC. Cumulative evidence suggests cancer stem cells (CSCs) are key drivers of tumor growth and heterogeneity. Post-transcriptional RNA editing of adenosine to inosine (A-to-I) catalyzed by ADAR1 dramatically alters cellular transcriptome in cancers. However, ADAR1 editase-dependent mechanisms governing hepatic CSC generation have not been elucidated. Therefore, we have systematically and comprehensively investigated ADAR1’s effect on CSC self-renewal in HCC. Methods: An in vitro human hepatocyte differentiation model along with relevant transcriptomic data form GEO and TCGA were analyzed to characterize the oncofetal role of ADAR1. Using genetic approaches, RNA sequencing was performed to identify putative ADAR1-mediated recoding edited genes in HCC cell lines. Clinical implication of GLI1 editing was studied in a cohort of 88 HCC patients. GLI1 editing-mediated change in its promoter activity and protein stability was investigated by dual reporter assay, CHIP, Co-IP, and PLA assays. Functional difference in stemness properties, including abilities of self-renewal, differentiation, tumorigenesis, chemo-resistance and metastasis between wild-type and edited GLI1 (GLI1wt vs. GLI1R701G) and the exact mechanisms were also studied in cell models and mice. Results: Lentiviral ADAR1 wild-type but not editing-defective ADAR1E912A mutant, editing at nt2101 of GLI1 transcript caused arginine-to-glycine substitution at the residue 701. Importantly, increased editing of GLI1 was implicated in the pathogenesis of HCC. Upon editing, C-terminal half of GLI1 harbored a lower susceptibility to the inhibition of SUFU, thus promoting its nuclear translocation and activation. Moreover, GLI1R701G appeared more stable than GLI1wt due to reduced formation of specific K63-GLI1 substrate and β-TrCP-GLI1 complex. Edited GLI1 was found to strongly enhance targeted activation of NANOG and SOX9, resulting in accumulation of hepatic CSCs population, hepatocarcinogenesis, sorafenib-resistance and metastasis. Additionally, GLI1 editing initiated mitophagy via PINK1/Parkin-dependent pathway. Impaired mitophagy effectively antagonized the functions of GLI1R701G on hepatic CSC self-renewal and overcome chemoresistance. Conclusion: The critical advance of this study is that ADAR1 editase activity drives GLI1-dependent maintenance of the hepatic CSC population. Our discovery of a pivotal ADAR1-GLI1-PINK1 self-renewal axis provides the first mechanistic link between RNA-editing-driven malignant progression and mitochondrial homeostasis. Thus, ADAR1 represents a unique therapeutic vulnerability in liver CSCs with active mitophagy mediated by edited GLI1. Citation Format: Jie Luo, Lanqi Gong, Yuma Yang, Jiao Huang, Xiaona Fang, Baifeng Zhang, Ying Tang, Beilei Liu, Ming Liu, Lu Bai, Victor Ho-Fun Lee, Xin-Yuan Guan. ADAR1-dependent RNA editing of GLI1 drives hepatocellular carcinoma stem cell self-renewal by initiating mitophagy [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 6064.

Cancer metastasis remains the key barrier for patient’s cure. Tumor metastasis initiating from a single disseminated cell into the invasive nodule in the foreign microenvironment is poorly understand. In this study, we established the intravenous injection pulmonary metastatic model followed by single-cell RNA sequencing of disseminating cells from mice lungs at different timepoints, we identified a cluster of SLPI-high esophageal squamous cell carcinoma (ESCC) cells that mediates the lung metastasis evolution and predicts worse overall survival of ESCC patients. GSEA analysis reveals that the SLPI-high subset is functionally associated with epithelial-to-mesenchymal transition and neutrophil activation during lung metastasis. Immune infiltration prediction from TIMER also confirmed that high SLPI is tightly correlated with more abundance of neutrophils in the ESCC tumor microenvironment. SLPI is reported to drive the metastasis progression through regulating the vascular mimicry. In the next step, further in-vitro and in-vivo studies is required to explore the role of SLPI in ESCC cells in promoting lung metastasis evolution. We identified a SLPI+ ESCC cells with high pluripotency that particularly initiates the metastasis process. Citation Format: Yu Zhang, Ching Ngar Wong, Lanqi Gong, Xin-yuan Guan, Beibei Ru, Beilei Liu. Single cell RNA sequencing uncovers high-plasticity cell state during tumor metastasis [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 5981.

Background and Purpose Our previous studies have shown that Src homology 2 (SH2) B adaptor protein 1 (SH2B1) plays an important role in cardiac hypertrophy, but the specific mechanism remains to be studied. Through bioinformatics and related research, it is found that miR-14 2-3 p is closely related to SH2B1. Exploring the relationship between miR-14 2-3 p and gene SH2B1 expression is beneficial for the treatment of cardiac hypertrophy. SH2B1 is a key factor regulating energy metabolism, mitochondria are the main organelles of energy metabolism and cardiac hypertrophy are closely related to mitochondrial dysfunction. So it is particularly important to explore the relationship between miR-14 2-3 p and SH2B1 and myocardial mitochondrial function. In this study, we investigated whether overexpression of miR-14 2-3 p can inhibit the expression of gene SH2B1, ameliorate cardiac mitochondrial dysfunction and cardiac hypertrophy. Methods We first constructed a pressure overload myocardial hypertrophy model by ligation of the abdominal aorta(AB) of rats. After 4 weeks of modeling, echocardiographic examination showed that the heart volume of the model group became larger, and Hematoxylin and Eosin Staining Kit (HE) staining showed that the cross-sectional area of the heart tissue became larger. The expression of atrial natriuretic peptide (ANP), brain natriuretic peptide (BNP), β-Myosin Heavy Chain (β-MHC) messenger RNA (mRNA) increased by real‑time polymerase chain reaction (PCR), which proved that the model of cardiac hypertrophy was successfully constructed. Then, miR-14 2-3 p agomir was injected into the tail vein of rats 2 weeks and 4 weeks respectively. The expression of miR-4 2-3 p mRNA was increased by PCR, suggesting that the miR-14 2-3 p plasmid was successfully transfected. At 4 weeks of pressure overload myocardial hypertrophy model, echocardiography was used to detect cardiac function. HE staining of heart tissue and the expression of ANP, BNP, β-MHC mRNA were used to detect cardiac hypertrophy. Flow cytometry was used to detect changes in mitochondrial membrane potential. Secondly, we observed the effect of miR-14 2-3 p on cardiomyocyte hypertrophy and mitochondrial function in vitro by culture neonatal rat cardiomyocytes. Afterwards, using angiotensin (Ang)II-, miRNA mimic- and miRNA mimic nc- treated cardiomyocytes for a given time. α-actin staining found that the myocardial cells became larger, The expression of ANP, BNP, β-MHC mRNA increased by PCR, which proved that AngII-induced cardiac hypertrophy was successfully constructed. Then, the mitochondrial density was measured using mitochondrial Mito-Red staining by Confocal microscope, the mitochondrial membrane potential was evaluated using flow cytometry, Mitochondrial respiration oxygen consumption rate (OCR) was measured by a Seahorse Extracellular Flux Analyzer XF96, and the expression levels of miR-14 2-3 p, ANP, BNP, β-MHC mRNA, SH2B1 in the cardiomyocytes of different groups were measured by RT-PCR and Western blotting. Finally, we used luciferase assay and transfected miR-14 2-3 p agomir in rats, transfected miR-14 2-3 p mimic in Cardiomyocytes, it is found that myocardial SH2B1 mRNA and protein expression both were reduced. Results When the pressure overload myocardial hypertrophy model was constructed for four weeks, echocardiography revealed that the heart volume, Left ventricular end diastolic diameter(LVIDd), Left ventricular end systolic diameter (LVIDs), Left ventricular posterior wall thickness (LVPWd), Systolic left ventricular posterior wall (LVPWs), Left ventricle (LV) Mass increased, Ejection fraction (EF) % decreased of AB group increased, but transfected with miR-14 2-3 p agomir of AB, these increase was not significant, EF% reduction was not obvious. HE staining showed that the myocardial cross-sectional area of AB group increased significantly, but the miR-14 2-3 p agomir treatment of AB group did not increase significantly. PCR analysis showed that the expression of ANP, BNP,β-MHC mRNA was significantly increased in AB group, but the miR-14 2-3 p agomir treatment of AB group was not significantly increased. Flow cytometry showed that the mitochondrial membrane potential of AB group was significantly reduced, and the miR-14 2-3 p agomir treatment of AB group was not significantly decreased. During AngII-induced cardiomyocyte hypertrophy, ANP, BNP,β-MHC mRNA expression was increased, while these factors was not significantly increased in miR-14 2-3 p mimic treatment group; mitochondrial membrane potential, mitochondrial density and OCR was significantly decreased in AngII treated group, and these were not significantly reduced in miR-14 2-3 p mimic treatment group; Conclusions miR-14 2-3 p not only mitigate cardiac hypertrophy by directly inhibit the expression of gene SH2B1, but also can protect mitochondrial function in cardiac hypertrophy of vitro and vivo.

Check dam is a line of defense in the comprehensive watershed management system, but with the extended operation time, the operational risk of check dam increases, coupled with the frequent occurrence of sudden heavy rainfall under the influence of climate change in recent years, further increasing the risk of dam break.In this study, to address the current problem of untimely detection of potential dam break risk of check dam in China, we use the common variable screening based on WoE (Weight of Evidence) and IV (Information Value), establish a score card model based on logistic regression method, use KS curves and statistics, and AUC values for model evaluation, and analyze the 2016 From 22:00 on August 16 to 12:00 noon on August 17, 2016, a dam break was formed by a large rainstorm in Dalat Banner.The following conclusions were obtained: The IV values of each variable, such as control area, total storage capacity, siltation area, storage volume, and maximum rainfall, were all greater than 0.1, and each variable had more than moderate predictive power as a function of the breaking of check dam, that is, each factor had an effect on the breaking of check dam.The IV values of precipitation factors are higher than those of check dam factors. The influence probability of precipitation factors on the breaking of check dam is about 67%, and the influence probability of check dam factors on the breaking of check dam is about 33%.Precipitation is a direct factor affecting the breaking of check dam, and the maximum 6-hour precipitation has the most significant effect; among the check dam factors, the control area, storage volume, and flood storage capacity have a greater effect on the breaking of check dam, accounting for more than 10%, while the siltation volume and depth have a smaller effect on the breaking of check dam, accounting for 2.85% and 2.89% respectively.Very low risk which check dam with a score of 95 or higher account for about 43%, low risk for 28%, average risk for 15%, and higher risk for 7%. Very high risk accounted for 5% and there was only one dam with danger which is rated below 10.This study aims to monitor and warn the operation of check dam to ensure the safe operation of the check dam system and maintain the ecological security of the Loess Plateau.

The tumor microenvironment (TME) of nasopharyngeal carcinoma (NPC) harbors a heterogeneous and dynamic stromal population. A comprehensive understanding of this tumor-specific ecosystem is necessary to enhance cancer diagnosis, therapeutics, and prognosis. However, recent advances based on bulk RNA sequencing remain insufficient to construct an in-depth landscape of infiltrating stromal cells in NPC. Here we apply single-cell RNA sequencing to 66,627 cells from 14 patients, integrated with clonotype identification on T and B cells. We identify and characterize five major stromal clusters and 36 distinct subpopulations based on genetic profiling. By comparing with the infiltrating cells in the non-malignant microenvironment, we report highly representative features in the TME, including phenotypic abundance, genetic alternations, immune dynamics, clonal expansion, developmental trajectory, and molecular interactions that profoundly influence patient prognosis and therapeutic outcome. The key findings are further independently validated in two single-cell RNA sequencing cohorts and two bulk RNA-sequencing cohorts. In the present study, we reveal the correlation between NPC-specific characteristics and progression-free survival. Together, these data facilitate the understanding of the stromal landscape and immune dynamics in NPC patients and provides deeper insights into the development of prognostic biomarkers and therapeutic targets in the TME., The tumor microenvironment can influence patient survival response to therapy. Here, the authors used single-cell sequencing to investigate the microenvironment of nasopharyngeal cancer and identify tumor-specific signatures in five stromal clusters of cells that may influence patient survival.

Customer citizenship behavior in the online shopping environment is vital to the success of e-retailers. However, it is unclear whether and how service recovery expectation and recovery justice predict customer citizenship behavior in e-retailing settings. Grounded on the expectation confirmation theory and social exchange theory, this study examined the influence of service recovery expectation and recovery justice on customer citizenship behavior with a serial mediation of recovery expectation confirmation and post-recovery satisfaction. A total of 774 samples from e-shoppers with most impressive (N = 401) and most recent (N = 373) service recovery experience were collected to test the hypotheses using structural equation modeling and bootstrapping procedures. This study reveals that service recovery expectation has a negative impact on recovery expectation confirmation, while recovery justice positively affects recovery expectation confirmation, which is further positively correlated with post-recovery satisfaction and customer citizenship behavior. Moreover, recovery expectation confirmation and post-recovery satisfaction play a serial mediation in the relationship between service recovery expectation and recovery justice, and customer citizenship behavior. Our study contributes to the growing body of customer citizenship behavior literature by offering an alternative perspective (i.e., service recovery) to understand what encourage or impede customer citizenship behavior, and expands service recovery literature by combining service recovery expectation and recovery justice into a framework and revealing the expectation–confirmation mechanism through which they influence post-recovery satisfaction in online shopping setting.

β-adrenergic activation is able to exacerbate cardiac hypertrophy. Myosin light chain kinase (MLCK) and its phosphorylated substrate, phospho-myosin light chain 2 (p-MLC2), play vital roles in regulating cardiac hypertrophy. However, it is not yet clear whether there is a relationship between β-adrenergic activation and MLCK in the progression of cardiac hypertrophy. Therefore, we explored this relationship and the underlying mechanisms in this work.Cardiac hypertrophy and cardiomyocyte hypertrophy were induced by pressure overload and isoproterenol (ISO) stimulation, respectively. Echocardiography, histological analysis, immunofluorescence and qRT-PCR were used to confirm the successful establishment of the models. A β-blocker (metoprolol) and a calpain inhibitor (calpeptin) were administered to inhibit β-adrenergic activity in rats and calpain in cardiomyocytes, respectively. The protein expression levels of MLCK, myosin light chain 2 (MLC2), p-MLC2, myosin phosphatase 2 (MYPT2), calmodulin (CaM) and calpain were measured using western blotting. A cleavage assay was performed to assess the degradation of recombinant human MLCK by recombinant human calpain.The β-blocker alleviated cardiac hypertrophy and dysfunction, increased MLCK and MLC2 phosphorylation and decreased calpain expression in pressure overload-induced cardiac hypertrophy. Additionally, the calpain inhibitor calpeptin attenuated cardiomyocyte hypertrophy, upregulated MLCK and p-MLC2 and reduced MLCK degradation in ISO-induced cardiomyocyte hypertrophy. Recombinant human calpain degraded recombinant human MLCK in vitro in concentration- and time-dependent manners, and this degradation was inhibited by the calpain inhibitor calpeptin.Our study suggested that β-adrenergic activation may promote the degradation of MLCK through calpain in pressure overload-induced cardiac hypertrophy.

High mobility group box-1 (HMGB1) participates actively in oxidative stress damage, and the latter relates closely to diabetes and diabetic complications including osteoporosis, though the underlying mechanisms are elusive. This study aimed to investigate the effect of high glucose on bone marrow stromal cells (BMSCs) apoptosis and the role of HMGB1 in this process. BMSCs were isolated from 2-week-old Sprague-Dawley rats and cultured in medium containing normal glucose (NG), high glucose (HG), high glucose + glycyrrhizin (HMGB1 inhibitor, HG+GL), and high glucose + glycyrrhizin + dorsomorphin (AMPK inhibitor, HG+GL+Dm), respectively. Cell apoptosis, expression of HMGB1, AMPK, apoptotic markers, and mitochondrial functions were detected. By these approaches, we demonstrated that HG treatment significantly upregulated the expression of HMGB1 in BMSCs, which could be attenuated by GL treatment. Inhibiting HMGB1 by GL improved AMPK activation, decreased mitochondrial ROS levels, increased mitochondrial membrane potential, normalized mitochondrial fission/fusion balance, and consequently reduced apoptosis of BMSCs under HG condition. The addition of AMPK inhibitor dorsomorphin hampered this protective effect. Taken together, our data show that inhibition of HMGB1 can be an effective approach to alleviate HG-induced BMSCs apoptosis by activation of AMPK pathway and relieving mitochondrial dysfunction.

Oxygen deficiency is associated with various oral diseases, including chronic periodontitis, age-related alveolar bone loss, and mechanical stress-linked cell injury from orthodontic appliances. Nevertheless, our understanding of the impact of hypoxia on periodontal tissues and its biochemical mechanism is still rudimentary. The purpose of this research was to elucidate the effects of hypoxia on the apoptosis of human periodontal ligament stem cells (PDLSCs) in vitro and the underlying mechanism. Herein, we showed that cobalt chloride (CoCl2) triggered cell dysfunction in human PDLSCs in a concentration-dependent manner and resulted in cell apoptosis and oxidative stress overproduction and accumulation in PDLSCs. In addition, CoCl2promoted mitochondrial fission in PDLSCs. Importantly, CoCl2increased the expression of dynamin-related protein 1 (Drp1), the major regulator in mitochondrial fission, in PDLSCs. Mitochondrial division inhibitor-1, pharmacological inhibition of Drp1, not only inhibited mitochondrial fission but also protected against CoCl2-induced PDLSC dysfunction, as shown by increased mitochondrial membrane potential, increased ATP level, reduced reactive oxygen species (ROS) level, and decreased apoptosis. Furthermore, N-acety-l-cysteine, a pharmacological inhibitor of ROS, also abolished CoCl2-induced expression of Drp1 and protected against CoCl2-induced PDLSC dysfunction, as shown by restored mitochondrial membrane potential, ATP level, inhibited mitochondrial fission, and decreased apoptosis. Collectively, our data provide new insights into the role of the ROS-Drp1-dependent mitochondrial pathway in CoCl2-induced apoptosis in PDLSCs, indicating that ROS and Drp1 are promising therapeutic targets for the treatment of CoCl2-induced PDLSC dysfunction.