Multi-Omics Integrated Analysis of the Protective Effect of EZH2 Inhibition in Mice with Renal Ischemia-Reperfusion Injury.

INTRODUCTION: Acute kidney injury (AKI) is a common clinical syndrome associated with high morbidity and mortality. Inhibition of the methyltransferase enhancer of zeste homolog 2 (EZH2) by its inhibitor 3-deazaneplanocin A (3-DZNeP) exerts renal benefits in acute renal ischemia-reperfusion injury (IRI). However, the underlying mechanisms are not completely known. This study aimed to elucidate the pathological mechanism of EZH2 in renal IRI by combination of multi-omics analysis and expression profiling in a public clinical cohort. METHODS: In this study, C57BL/6 J mice were used to establish the AKI model, which were treated with 3-DZNeP for 24 h. Kidney samples were collected for RNA-seq analysis, which was combined with publicly available EZH2 chromatin immunoprecipitation sequencing (ChIP-seq) data of mouse embryonic stem cell for a joint analysis to identify differentially expressed genes. Several selected differentially expressed genes were verified by quantitative PCR. Finally, single-nucleus sequencing data and expression profiling in public clinical datasets were used to confirm the negative correlation of the selected genes with EZH2 expression. RESULTS: 3-DZNeP treatment significantly improved renal pathology and function in IRI mice. Through RNA-seq analysis combined with EZH2 ChIP-seq database, 162 differentially expressed genes were found, which might be involved in EZH2-mediated pathology in IRI kidneys. Four differential expressed genes (Scd1, Cidea, Ghr, and Kl) related to lipid metabolism or cell growth were selected based on Gene Ontology and Kyoto Encyclopedia of Genes and Genome enrichment analysis, which were validated by quantitative PCR. Data from single-nucleus RNA sequencing revealed the negative correlation of these four genes with Ezh2 expression in different subpopulations of proximal tubular cells in IRI mice in a different pattern. Finally, the negative correlation of these four genes with EZH2 expression was confirmed in patients with AKI in two clinical datasets. CONCLUSIONS: Our study indicates that Scd1, Cidea, Ghr, and Kl are downstream genes regulated by EZH2 in AKI. Upregulation of EZH2 in AKI inhibits the expression of these four genes in a different population of proximal tubular cells to minimize normal physiological function and promote acute or chronic cell injuries following AKI.

Biomarker discovery and application-An opportunity to resolve the challenge of liver cancer diagnosis and treatment.

Liver cancer is one of the most common malignancies, with severe morbidity and mortality. While considerable progress has been made in liver cancer treatment, the 5-year overall survival (OS) of patients has not improved significantly. Reasons include the inadequate capability of early screening and diagnosis, a high incidence of recurrence and metastasis, a high degree of tumor heterogeneity, and an immunosuppressive tumor microenvironment. Therefore, the identification and validation of specific and robust liver cancer biomarkers are of major importance for early screening, timely diagnosis, accurate prognosis, and the prevention of tumor progression. In this review, we highlight some of the latest research progress and potential applications of liver cancer biomarkers, describing hotspots and prospective directions in biomarker discovery.

Additively Manufactured Multilevel Voronoi-Lattice Scaffolds with Bonelike Mechanical Properties.

Irregular porous scaffold through Voronoi tessellation based on global modeling demonstrated randomness to a certain degree and susceptibility to producing large processing deviations. A modeling method for new types of scaffolds based on periodic arrays of Voronoi unit cell was proposed in this study. These porous scaffolds presented controllable local cells and satisfactory mechanical properties. The topological structure of the Voronoi unit cell was controlled using three independent cell design factors (Voronoi polyhedron volume V, face-centered scaled factor F1, and body-centered scaled factor F2), and multilevel Voronoi-lattice scaffolds were constructed on the basis of periodic arrays of the Voronoi unit cell. Compressive test and simulation were combined to quantify the mechanical properties of scaffolds. The regression equations were established using the response surface method (RSM) to determine relationships between Voronoi unit cell design factors and structural characteristic parameters and mechanical properties. The same trends were observed in stress-strain curves of the compressive test and simulation. The mechanical properties of scaffolds can be appropriately quantified via simulation. Regression equations based on RSM can properly predict the structural characteristic parameters and mechanical properties of the scaffold. Compared with V, F1 and F2 exerted a stronger influence on the structural characteristic parameters and mechanical properties of the scaffold. The modeling method of the multilevel Voronoi-lattice scaffold based on the Voronoi unit cell was proposed in this study to design the porous scaffold and meet the requirements of human bone morphology, mechanical properties, and actual manufacturing by adjusting factors V, F1, and F2. The proposed method can provide a feasible strategy for designing implants with suitable and similar morphologies and mechanical properties to cancellous bone.

Chiral rhodium(III)-azobenzene complexes as photoswitchable DNA molecular locks.

Chiral rhodium(III)-azobenzene complexes that are able to intercalate into DNA were developed. Upon light exposure, the azobenzene moiety of the metal complexes can photoisomerize from the trans-form to the cis-form, and strongly stabilize the DNA double-helix and modulate DNA transcription. This study presents the first example of metal-based photoswitchable DNA molecular locks.

[Design of new gradient scaffolds based on triply periodic minimal surfaces and study on its mechanical, permeability and tissue differentiation characteristics].

In order to establish a bone scaffold with good biological properties, two kinds of new gradient triply periodic minimal surfaces (TPMS) scaffolds, i.e., two-way linear gradient G scaffolds (L-G) and D, G fusion scaffold (N-G) were designed based on the gyroid (G) and diamond (D)-type TPMS in this study. The structural mechanical parameters of the two kinds of scaffolds were obtained through the compressive simulation. The flow property parameters were also obtained through the computational fluid dynamics (CFD) simulation in this study, and the permeability of the two kinds of scaffolds were calculated by Darcy's law. The tissue differentiation areas of the two kinds of scaffolds were calculated based on the tissue differentiation theory. The results show that L-G scaffold has a better mechanical property than the N-G scaffold. However, N-G scaffold is better than the L-G scaffold in biological properties such as permeability and cartilage differentiation areas. The modeling processes of L-G and N-G scaffolds provide a new insight for the design of bone scaffold. The simulation in this study can also give reference for the prediction of osseointegration after the implantation of scaffold in the human body.

Ruthenium(II) complexes as bioorthogonal two-photon photosensitizers for tumour-specific photodynamic therapy against triple-negative breast cancer cells.

Herein, we developed the first Ru(ii) complex-based bioorthogonal two-photon photosensitizers. Through bioorthogonal labelling, they realize effective tumour-specific photodynamic therapy against triple-negative breast cancer cells.

Influence of postoperative adjuvant transarterial chemoembolization on the prognosis of early-stage intrahepatic cholangiocarcinoma: a single center study.

BACKGROUND: The effectiveness of postoperative adjuvant transarterial chemoembolization (TACE) on survival and recurrence in tumor-node-metastasis (TNM) stage I intrahepatic cholangiocarcinoma (ICC) after radical resection remains unclear. This study aimed to compare overall survival (OS) and recurrence-free survival (RFS) in TNM stage I ICC patients with and without postoperative TACE. METHODS: A retrospective cohort study was conducted on TNM stage I ICC patients who had undergone R0 resections with curative intent in Shanghai Eastern Hepatobiliary Surgery Hospital from January 2012 to December 2016. A total of 269 patients were divided into two groups: (I) 35 patients who received postoperative TACE and (II) 234 patients no TACE. Staging was performed according to the 8th edition of American Joint Committee on Cancer (AJCC) TNM staging system. The tumor-related RFS and OS were estimated by the Kaplan-Meier method. Cox proportional regression model was employed to evaluate the prognosis between the two groups. RESULTS: In all patients, the median OS was 66.8 months. After R0 resection, adjuvant TACE could not improve the survival of TNM stage I patients, and the OS of the TACE group was not better than that of the non-TACE group (P=0.7070). In addition, in the TACE group, the recurrence rate of TNM stage I ICC patients was statistically significantly higher than that of the non-TACE group (P=0.0328). Multivariable analysis revealed that adjuvant TACE was an independent predictor of worse RFS (HR: 1.88, 95% CI: 1.21-2.93). CONCLUSIONS: Adjuvant TACE after radical surgery failed to prolong the OS and potentially delay recurrence for patients with TNM Stage I ICC. Adjuvant TACE might not be suitable for patients with TNM Stage I ICC.

Intrinsic strategies for the evasion of cGAS-STING signaling-mediated immune surveillance in human cancer: How therapy can overcome them.

Cyclic GMP-AMP synthase (cGAS) recognizes cytosolic DNA and catalyzes the formation of cyclic GMP-AMP, which upon activation triggers the induction of stimulator of interferon genes (STING), leading to type I interferons production; these events then promote the cross-priming of dendritic cells and the initiation of a tumor-specific CD8+ T cell response. However, cancer cells in the tumor microenvironment cannot trigger intrinsic cGAS-STING signaling, regardless of the expression of cGAS and STING. This dysfunctional cGAS-STING signaling enables cancer cells to evade immune surveillance, thereby promoting tumorigenesis. Here, we review recent advances in the current understanding of the activation of cGAS-STING signaling and immunotherapies based on this pathway and focus on the mechanisms for the inactivation of this pathway in tumor cells to promote the development of cancer immunotherapy. The discovery of inherent resistance and the selection of appropriate combination therapies are of great significance for tumor treatment development.

Human Cancer Cells Sense Cytosolic Nucleic Acids Through the RIG-I-MAVS Pathway and cGAS-STING Pathway.

Pattern recognition receptors (PRRs) are germline-encoded host sensors of the innate immune system. Some human cancer cells have been reported to express PRRs. However, nucleic acid sensors in human cancers have not been studied in detail. Therefore, we systematically analyzed the expression, molecular cascade, and functions of TLR3, RIG-I, MDA5, LGP2, cGAS, and STING in human cancer cells. TLR3, TRIF, RIG-I, MDA5, LGP2, and MAVS were expressed in 22 cell lines. The majority of cell lines responded to only RIG-I ligands 5'-ppp-dsRNA, Poly(I:C)-HMW, Poly(I:C)-LMW, and/or Poly(dA:dT), as revealed by IRF3 phosphorylation and IFN-ß secretion. IFN-ß secretion was inhibited by RIG-I and MAVS knockdown. cGAS and STING were co-expressed in 10 of 22 cell lines, but IFN-ß secretion was not induced by STING ligands ISD, HSV60, VACV70, Poly(dG:dC), and 3'3'-cGAMP in cGAS and STING intact cell lines. Further experiments revealed that the cGAS-STING pathway was activated, as revealed by TBK1 and IRF3 phosphorylation and IFN-ß and ISG mRNA expression. These results suggest that human epithelial cancer cells respond to cytosolic RNA through the RIG-I-MAVS pathway but only sense cytosolic DNA through the cGAS-STING pathway. These findings are relevant for cancer immunotherapy approaches based on targeting nucleic acid receptors.

Desflurane and Surgery Exposure During Pregnancy Decrease Synaptic Integrity and Induce Functional Deficits in Juvenile Offspring Mice.

Anesthesia in pregnant women may cause adverse effects in the hippocampus of unborn babies and fetal brain development. The mechanisms underlying pathological changes resulting from anesthetics are unclear. This study tested the hypothesis that exposure to desflurane during pregnancy may impair cognition and memory functions of juvenile offspring. Pregnant mice (at gestational day 14) were administered 10% desflurane for 3 h and compared to sham control and sciatic nerve hemi-transection surgery. Hippocampal tissues of both fetal (G14) and offspring mice (postnatal day 31) were collected and analyzed by real-time qPCR and Western blot. Functional tests were performed to assess fear and memory functions in offspring mice. Primary hippocampal neuronal cultures from postnatal day 0 (without desflurane exposure) were examined for neuronal and synaptic development under desflurane treatment in vitro. In this acute experiment, we showed that neuronal cultures exposed to desflurane significantly increased interleukin (IL)-6 expression and apoptotic gene caspase-3 activation. Desflurane exposure significantly reduced PSD-95 expression in hippocampal neurons. Similar changes were observed in hippocampal tissues from juvenile offspring mice. Inhaled desflurane impaired memory functions in offspring mice compared to sham control. These mice displayed higher sensitivity to fear conditioning. Neurons isolated from the mice exposed to desflurane exhibited significantly lower levels of synaptophysin expression. These results suggest that anesthetic exposure together with surgery during pregnancy may induce detrimental effects in juvenile offspring mice via the induction of cell death and disruption of synaptic integrity.

Optimization of extraction of polysaccharide from dandelion root by response surface methodology: Structural characterization and antioxidant activity.

In this study, response surface methodology (RSM) was used to optimize the extraction parameters of dandelion root polysaccharide (DRP). Two novel polysaccharides (DRP-2b, DRP-3a) were isolated from dandelion root by graded ethanol precipitation and column chromatography. Structural analyses indicated that DRP-2b, with a molecular weight of 31.8 kDa, was composed of rhamnose, glucuronic acid, glucose, galactose and arabinose, whereas DRP-3a, with a molecular weight of 6.72 kDa, was composed of rhamnose, glucose, galactose and arabinose. The backbones of DRP-2b and DRP-3a were mainly composed of (1 → 5)-linked-α-D-Ara and (1 → 6)-linked-α-D-Glc, respectively. In addition, the antioxidant activities and protective effects against H2O2-induced damage in hepatic L02 cells were evaluated. DRP-3a exhibited higher radical scavenging activity than DRP-2b against the DPPH, hydroxyl and superoxide anions. Furthermore, DRP-3a exhibited strong protective effects against H2O2-induced damage in hepatic L02 cells. Thus, DRP-3a could be used as a potential antioxidant in medicine or as a functional food.

The Role of Long Noncoding RNAs in Central Nervous System and Neurodegenerative Diseases.

Long noncoding RNAs (lncRNAs) refer to a group of noncoding RNAs (ncRNAs) that has a transcript of more than 200 nucleotides in length in eukaryotic cells. The lncRNAs regulate gene expression at epigenetic, transcriptional, and post-transcriptional levels by multiple action modes. In this review, we describe the diverse roles reported for lncRNAs, and discuss how they could mechanistically be involved in the development of central nervous system (CNS) and neurodegenerative diseases. Further studies on the function of lncRNAs and their mechanism will help deepen our understanding of the development, function, and diseases of the CNS, and provide new ideas for the design and development of some therapeutic drugs.

Structural characterization, antioxidant and hepatoprotective activities of polysaccharides from Sophorae tonkinensis Radix.

In current study we present two polysaccharides, STRP1 and STRP2, purified from Sophorae tonkinensis Radix via column chromatography. Structural analyses indicated that STRP1 and STRP2 were consisted of mannose, rhamnose, glucuronic acid, glucose, galactose and arabinose in a similar molar ratio with main backbones of (1 → 3)-linked-α-d-Gal and (1 → 4)-linked-α-d-Glc, while average molecular weights were 1.30 × 104 and 1.98 × 105 Da, respectively. We observed a strong chelating ability on ferrous ions; substantial radical scavenging activities on DPPH, hydroxyl and superoxide anion radicals in vitro; and significant attenuation on acetaminophen-induced hepatic oxidative damage in mice for STRP1 and STRP2. The promising data on these polysaccharides showcase the need to further develop novel natural antioxidant and liver-protecting drugs.

Interfering with DNA High-Order Structures using Chiral Ruthenium(II) Complexes.

In this work, it was found that DNA can undergo B-Z transformational changes and compaction in the presence of DNA intercalators such as ruthenium(II) polypyridyl complexes. The link between B-Z transition and condensation is weak but can be strengthened under certain circumstances with slight alterations to the structures of the ruthenium(II) complexes. Here, following on from previous research, this work reports a series of ruthenium(II) complexes with imidazophenanthroline ligands, which vary in size and planarity. The complexes exhibit distinct effects on DNA structures, ranging from little impact to the transformation of DNA secondary structures to the formation of higher-order DNA structures. Further studies on DNA morphological changes induced by chiral ruthenium(II) complexes are observed by atomic force microscopy and transmission electron microscopy.

Research progress on the roles of microRNAs in governing synaptic plasticity, learning and memory.

The importance of non-coding RNA involved in biological processes has become apparent in recent years and the mechanism of transcriptional regulation has also been identified. MicroRNAs (miRNAs) represent a class of small regulatory non-coding RNAs of 22bp in length that mediate gene silencing by identifying specific sequences in the target messenger RNAs (mRNAs). Many miRNAs are highly expressed in the central nervous system in a spatially and temporally controlled manner in normal physiology, as well as in certain pathological conditions. There is growing evidence that a considerable number of specific miRNAs play important roles in synaptic plasticity, learning and memory function. In addition, the dysfunction of these molecules may also contribute to the etiology of several neurodegenerative diseases. Here we provide an overview of the current literatures, which support non-coding RNA-mediated gene function regulation represents an important but underappreciated, layer of epigenetic control that facilitates learning and memory functions.

Differentially expressed lncRNAs and miRNAs with associated ceRNA networks in aged mice with postoperative cognitive dysfunction.

Postoperative cognitive dysfunction (POCD) is a common postoperative complication observed in elderly patients. Using microarray analyses, we comprehensively compared long non-coding RNA (lncRNA), messenger RNA (mRNA), and microRNA (miRNA) expression profiles in hippocampal tissues from a mouse model of POCD and control mice. A total of 175 lncRNAs, 117 mRNAs, and 26 miRNAs were differentially expressed between POCD and control mice. Gene ontology (GO) and KEGG pathway enrichment analyses were performed to explore the principal functions of dysregulated genes. Correlated coding-noncoding co-expression (CNC) and competing endogenous RNA (ceRNA) expression networks were constructed using bioinformatics methods. lncRNA NONMMUT000708 correlated positively with expression of the inflammation-related gene Hif3a. lncRNAs NONMMUT043249 and NONMMUT028705 mediated gene expression by binding the transcription factor cAMP response element-binding protein (CREB). The constructed ceRNA network suggested lncRNA NONMMUT055714 binds competitively with miR-7684-5p, increasing expression of its target gene, Sorl1. Finally, eight dysregulated lncRNAs, four miRNAs, and ten mRNAs were confirmed via quantitative real-time polymerase chain reaction (PCR) in 10 POCD-healthy mouse paired samples. These results suggest that lncRNAs and miRNAs are involved in POCD pathogenesis and progression. Our ceRNA network will improve understanding of lncRNA-mediated ceRNA regulatory mechanisms operating during the pathogenesis of POCD.

CYP2A6 Polymorphisms Associate with Outcomes of S-1 Plus Oxaliplatin Chemotherapy in Chinese Gastric Cancer Patients.

Gastric carcinoma is a heterogeneous malignant disease involving genetic factors. To identify predictive markers for gastric cancer treatment in Chinese patients, we evaluated the association between polymorphisms of the gene encoding cytochrome P450 2A6 (CYP2A6) and outcomes of S-1 plus oxaliplatin (SOX) chemotherapy treatment. Clinical data on 60 consecutive gastric cancer patients receiving SOX regimen were collected prospectively. We sequenced all exons of CYP2A6 and a total of 22 different polymorphisms were detected in the present study. Comprehensive analyses of these genetic polymorphisms were performed to determine their association with both safety and efficacy of SOX regimen. Our results showed that polymorphisms of CYP2A6 were associated with the safety and efficacy of SOX treatment. Among them, missense mutations CYP2A6 rs60823196 and rs138978736 could be possible risk factors (P<0.05) for severe diarrhea induced by SOX, whereas CYP2A6 rs138978736 could be a conceivable predictor for overall survival of patients treated with SOX adjuvant chemotherapy. Further large-scale randomized prospective studies are warranted to confirm these findings.

MiR-7684-5p leads to surgery-induced cognitive decline in mice probably through the downregulation of SorLA.

BACKGROUND: Postoperative cognitive dysfunction is a postoperative severe complication caused by many factors. However, its specific pathogenesis remains unclear. MicroRNAs (miRNAs), which are involved in the pathogenesis of neurodegenerative diseases, may also affect POCD. METHODS: In this research, microarray technology was used to screen 26 miRNAs that had a differential expression in the hippocampus of mouse between the surgery group and control group. The qRT-PCR verification on the hippocampuses of 10 pairs of mouse testifies the high expression of miR-7684-5p in the surgery group (identical with the result of chip). RESULTS: Surgical trauma was found to induce the expression of miR-7684-5p with the accumulation of Aß in the hippocampus. Furthermore, miR-7684-5p knockdown effectively reduced the levels of Aß triggered by surgery, and attenuated hippocampal-dependent memory impairment. Moreover, we testify that sorLA is a target gene of miR-7684-5p through bioinformatics prediction and dual-luciferase report gene experiment. CONCLUSIONS: Our data indicate that decreased postoperative cognitive function may be caused by the increased generation of Aß by reducing sorLA expression. Our work implicates miR-7684-5p as a potential biomarker and a novel therapeutic target.

UGT1A1*6, UGT1A7*3 and UGT1A9*1b polymorphisms are predictive markers for severe toxicity in patients with metastatic gastrointestinal cancer treated with irinotecan-based regimens.

Irinotecan-induced severe neutropenia and diarrhea, which remain unpredictable, has restrained the dose and clinical efficiency of irinotecan administration. In the present study, a total of 70 irinotecan-treated patients with histologically confirmed metastatic gastrointestinal cancer were enrolled. Despite genotyping well-reported alleles, direct sequencing was specifically adopted to avoid ethnic heterogeneity and to identify novel variations. The promoter (-1000 bp) and exon 1 regions of UDP glucuronosyltransferase family 1 member A complex locus (UGT1A1) gene family members UGT1A1, UGT1A7 and UGT1A9 were sequenced, and comprehensive analysis of their genetic polymorphisms was performed to determine the association between inherited genetic variations and irinotecan-induced toxicity. A total of 23 different genetic variants were detected in the present study, including 2 novel polymorphisms. The results of the present study revealed that UGT1A1*6 and UGT1A7*3 are risk factors for irinotecan-induced severe neutropenia, and UGT1A9*1b is associated with severe diarrhea. These results may provide biomarkers for the selection of the optimal chemotherapy for Chinese patients with metastatic gastrointestinal cancer.

The identification of human aldo-keto reductase AKR7A2 as a novel cytoglobin-binding partner.

Cytoglobin (CYGB), a member of the globin family, is thought to protect cells from reactive oxygen and nitrogen species and deal with hypoxic conditions and oxidative stress. However, its molecular mechanisms of action are not clearly understood. Through immunoprecipitation combined with a two-dimensional electrophoresis-mass spectrometry assay, we identified a CYGB interactor: aldo-keto reductase family 7 member A2 (AKR7A2). The interaction was further confirmed using yeast two-hybrid and co-immunoprecipitation assays. Our results show that AKR7A2 physically interacts with CYGB.