Emerging evidence indicates that circadian rhythms profoundly influence the pathophysiology of heart failure (HF). The circadian clock system governs myocardial functions via synchronized actions from multiple cell types involving cardiomyocytes and non-cardiomyocytes. This review summarizes state-of-the-art advances on the cardiovascular circadian system and provides a perspective on time-of-day effects in the pathophysiology of HF, including energy metabolism, tissue inflammation and fibrosis. In particular, we focus on cell-specific circadian regulation and synthesize how circadian clocks across cardiomyocytes, vascular cells, fibroblasts, and adipocytes interact to regulate the pathophysiology of HF. Based on this framework, we discuss how synchronizing internal and external time cues via clock- modulating compounds and/or time-restricted eating may prevent HF or facilitate rehabilitation. Together, this Review provides a timely synthesis of current understanding of HF as a function of biological timing and highlights potential chronotherapeutic strategies.

Background Oligoasthenoteratospermia (OAT) represents a major cause of male infertility. The molecular mechanisms underlying sperm morphological defects, particularly their potential interplay with environmental exposure-associated molecular alterations, remain poorly understood and warrant exploratory investigation. Methods Semen and plasma samples were collected from normal controls (NC, n = 6), mild morphological defect (MMD, n = 6), and severe morphological defect (SMD, n = 6) subjects in this exploratory pilot study designed to detect large-effect signals. Integrated proteometabolomic profiling was performed using 4D proteomics and LC-MS. Bioinformatic analysis focused on screening differential expression, functional enrichment, exploring potential interactions between exposure-associated metabolites and proteins, and identifying coherent patterns to potentially differentiate groups. Results Comparative analysis identified biphasic dysregulation patterns in key cytoskeletal proteins. The prominent hub ACTN4 exhibited depletion in MMD sperm while displaying pathological spatial dislocation with 83% showing midpiece signal loss and fragmented tail fluorescence, accompanied by extracellular vesicle release in SMD sperm. Quantitative immunofluorescence validation confirmed these divergent phenotypes, that is a 56% systemic reduction in MMD compared to controls (p < 0.0001), with SMD sperm also showing 41% absolute depletion versus NC (p < 0.0001) despite proteomic upregulation, which indicates functional loss through vesicular shedding. Plasma metabolomics revealed distinct profiles between severity groups, quantifying 357 differentially abundant metabolites (including prioritized selenometabolite-bile acid interactions) between SMD and MMD. Exploratory correlation analysis prioritized inverse associations including arsenocholine (an established environmental arsenic exposure biomarker) and ACTN4 (rho = -0.867, p = 0.0025), glycodeoxycholic acid versus PPP1CC (rho = -0.917, p < 0.001), and Se-adenosylselenomethionine-PGK1 (rho = -0.867, p = 0.0025), reflecting ultrastructural damage patterns. Conclusion This severity-stratified analysis demonstrates that sperm morphological defects in OAT associate with compartment-specific ACTN4 depletion patterns, that is quantitative loss systemically impairing head-tail coupling in MMD versus spatial dislocation with pathological vesiculation causing functional nullification in SMD. The exploratory findings suggest potential associations with multisystem exposure effects, particularly indicated by the arsenocholine-ACTN4 correlation, though causal relationships cannot be established from this observational study. In addition, exposure-protein associations involving PPP1CC and PGK1 also support the hypothesis of environmental perturbation affecting sperm integrity, though these observations require validation in independent cohorts.

Artificial light at night (ALAN) is an escalating environmental stressor linked to cardiovascular disease, yet its underlying mechanisms remain poorly understood. Here, we investigate the molecular basis of this pathology using zebrafish embryos for mechanistic discovery and murine models for translational validation under a chronic dim-light-at-night (dLAN) paradigm. dLAN exposure induced cardiac injury characterized by pericardial edema and hemodynamic impairment. Transcriptomic profiling revealed dysregulation of inflammatory and oxidative stress pathways, accompanied by marked suppression of the calcium-activated chloride channel accessory protein zclca1. Genetic knockdown ofzclca1 recapitulated core dLAN pathologies, while pharmacological intervention with the chloride-modulating diuretic bumetanide reversed cardiac dysfunction in both species. These findings identify a conserved "dLAN-CLCA-chloride-cardiac injury" axis, wherein light pollution disrupts chloride homeostasis to drive cardiac pathology, suggesting that chloride modulation may represent a potential therapeutic strategy for mitigating light pollution-induced cardiovascular injury.

Stress-induced gastric ulcers (SGU) are categorized as a stress-related mucosal disease (SRMD) that exhibits a high morbidity among ICU patients. However, due to the harmful side effects of proton pump inhibitors (PPIs), there is an urgent need for natural and proactive alternative methods in the treatment of SGU. Acacetin, a natural flavonoid, has demonstrated potential in alleviating gastric ulcers. To elucidate the effects of acacetin in SGU, in this study, gastric ulcers (GU)-related targets were retrieved from GeneCards, DisGeNET, and OMIM. A protein-protein interaction (PPI) network was constructed, followed by functional enrichment analyses, including Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways. Molecular docking was employed to predict the interactions between acacetin and key target proteins. Furthermore, a cold water-immersion restraint stress (CWIR) induced mouse model was used to validate the protective effects of acacetin on SGU. Network pharmacology and molecular docking revealed that acacetin exerts mucosal protective effects through multiple ferroptosis-related targets. In vivo, acacetin significantly mitigated the loss and disorganization of epithelial cells by inhibiting oxidative stress and ferroptosis, indicating its potential to protect against CWIR-related SGU. Especially, 25 mg/kg/day might be the optimal therapeutic dosage of acacetin as an alternative treatment option for SGU.

The circadian regulation of autophagy in airway epithelial inflammation remains poorly understood, particularly in the context of asthma. This study investigated how nuclear receptor subfamily 1 group D member 1 (NR1D1)-mediated downregulation of myosin-like BCL2-interacting protein (BECLIN1) influences autophagy in airway epithelial cells to alleviate inflammation. An ovalbumin-induced asthma mouse model and a Beas-2b cell line with type 2 inflammation were established. Rhythmic fluctuations in circadian clock genes (Circadian locomotor output cycles Kaput [Clock], Brain and muscle Arnt-like protein 1 [Bmal1], and Nr1d1) and autophagy-related genes (Autophagy Related 5 [Atg5], Beclin1, and microtubule-associated proteins 1A/1B light chain 3B [Lc3]) were monitored. In vitro, NR1D1 expression was modulated using SR9009 or siRNA, and effects on BECLIN1 and LC3 were assessed. BECLIN1 expression was altered via plasmid transfection to evaluate effects on NR1D1. NR1D1 bonded with and repressed the BECLIN1 promoter. In asthmatic mice and Beas-2b cells, NR1D1 expression was reduced, whereas ATG5, BECLIN1, and LC3 were elevated. CLOCK, BMAL1, NR1D1, ATG5, BECLIN1, and LC3 levels in the lung tissue showed cyclic fluctuations, with NR1D1 and BECLIN1 negatively correlated. SR9009 reduced BECLIN1 and LC3 levels, while NR1D1 knockdown increased them in vitro. BECLIN1 overexpression elevated NR1D1, whereas its inhibition reduced NR1D1. Circadian and autophagy-related genes showed pronounced periodic oscillation in asthmatic mice, coinciding with airway inflammation. NR1D1 negatively regulates BECLIN1 to suppress autophagy and mitigate airway inflammation in asthma, indicating a possible therapeutic target.

Accumulating evidence has indicated that lung ferroptosis is an important contributor to septic acute lung injury (SALI). Inducible nitric oxide synthase (iNOS) may be implicated in the regulation of bronchial epithelial ferroptosis. Nevertheless, the precise mechanisms by which iNOS modulates ferroptosis remain elusive. This study investigated whether iNOS selective inhibitor 1400w alleviates LPS-induced SALI and suppresses ferroptosis in mice. Additionally, RNA sequencing (RNA-seq), molecular docking, molecular dynamic simulation, Transmission electron microscope (TEM), and western blotting were employed to predict and evaluate the molecular mechanism of 1400w on LPS-induced ferroptosis in vivo. The results showed that the administration of 1400w markedly attenuated LPS-induced lung injury and facilitated pulmonary function in mice. Also, 1400w administration effectively suppressed bronchial epithelial ferroptosis induced by LPS in mice. Furthermore, molecular docking and molecular dynamics simulations revealed stable binding between GPX4 and iNOS, with 1400w modulating ferroptosis mediated by SLC7A11/GPX4 through targeting iNOS. Collectively, our research demonstrated that inhibition of iNOS might represent a potential therapeutic strategy to improve SALI by inhibiting ferroptosis.

A disrupted circadian rhythm can cause liver and intestinal diseases, however, the mechanisms remain unclear. Therefore, we examined the effects of the gut-liver axis circadian locomotor output cycles kaput (Clock) gene, on mouse liver and intestinal to identify how Clock regulates gut-liver axis functions. Wild-type (WT) and gut-liver axis-specific Clock knockdown (Clock-KD) mice were fed a normal diet. Intestinal microflora diversity and species abundance changes were analysed by 16s rDNA sequencing. A Caco2 cell monolayer model was established and Clock-KD effects was examined following Clock small-hairpin RNA transfection. Clock-KD mouse livers were not significantly changed. However, microflora abundance and biodiversity in these mice were significantly lower when compared with WT mice. Moreover, Clock was shown to regulate intestinal tight junction protein expression both in vivo and in vitro. Interestingly, the CLOCK protein interacted with TJP1. Clock affects intestinal flora diversity and composition and regulates intestinal barrier integrity in mice.

The increased global prevalence of metabolic dysfunction-associated steatohepatitis (MASLD) has been closely associated with chronic disorders of the circadian clock. Herein, we investigate the role of Clock, a core circadian gene, in the pathogenesis of MASLD. Wild-type (WT) and liver-specific Clock knockdown (Clock-KD) mice were fed a Western diet for 20 weeks to induce MASLD. A cellular MASLD model was established by treating AML12 cells with free fatty acids and the effects of Clock knockdown were examined following transfection with Clock siRNA. Increased lipid deposition and more severe steatohepatitis and fibrosis were observed in the livers of Western diet-fed but not normal chow diet-fed Clock-KD mice after 20 weeks compared to WT mice. Moreover, the Clock gene was found to be significantly downregulated in WT MASLD mice. The Clock gene was shown to regulate the expression of lipophagy-related proteins (LC3B, P62, RAB7, and PLIN2) in vivo and in vitro. Knockdown of Clock was found to inhibit lipophagy resulting in increased accumulation of lipid droplets in the mouse liver and AML12 cells. Interestingly, the CLOCK protein was shown to interact with P62. However, knockdown of the Clock gene did not promote transcription of the P62 gene but suppressed degradation of the P62 protein during lipophagy in AML12 cells. The hepatic Clock gene regulates lipophagy and affects lipid droplet deposition in liver cells, and thus plays a critical role in the development of MASLD induced by a Western diet.

Background:The testicular sperm instead of ejaculated sperm for intracytoplasmic sperm injection (ICSI) in infertile men with high sperm DNA fragmentation (SDF) is a controversial topic. This updated systematic review and meta-analysis aims to evaluate whether couples with high level of SDF will benefit more from intracytoplasmic sperm injection with testicular sperm (Testi-ICSI) as compared to intracytoplasmic sperm injection with ejaculated sperm (Ejac-ICSI).Methods:A systematic search was conducted according to PRISMA guidelines, using PubMed, Embase, Web of Science and the Cochrane Central Register of Controlled Trials (CENTRAL), encompassing studies from the earliest record until May 2022. We included studies analyzing comparative pregnancy outcomes of testicular versus ejaculated sperm for ICSI in infertile men with high DNA fragmentation. The risks of bias and certainty of evidence were assessed using the Risk Of Bias In Non-randomized Studies of Interventions (ROBINS-I) and the Grading of Recommendations Assessment, Development, and Evaluation (GRADE) framework, respectively.Results:Eleven studies were included. Meta-analysis showed that SDF levels revealed a significant difference association [odds ratio (OR) =-25.81; 95% confidence interval (CI): -34.82, -16.81; I2=94%; P<0.00001] between testicular and ejaculated sperm. Compared with Ejac-ICSI, a non-significant tendency was observed for fertilization rates (FRs) in the Testi-ICSI group (OR =0.87; 95% CI: 0.67, 1.12; I2=81%; P=0.28). However, there was significant difference pointing to better outcomes for Testi-ICSI in clinical pregnancy rates (CPRs) (OR =2.36; 95% CI: 1.71, 3.24; I2=0%; P<0.00001), live birth rates (LBRs) (OR =3.10; 95% CI: 2.13, 4.51; I2=4%; P<0.00001) and miscarriage rates (MRs) (OR =0.28; 95% CI: 0.13, 0.60; I2=0%; P=0.001).Conclusions:Results of this updated meta-analysis reveal that SDF rates are lower in testicular sperm than in ejaculated sperm and that Testi-ICSI is correlated with better clinical outcomes, including higher CPRs, higher LBRs, and lower MRs in infertile males with high SDF levels. Nevertheless, with the overall low to moderate quality of the studies, further well-designed controlled studies are required.

Background Polycystic ovary syndrome (PCOS) is one of the most common endocrine abnormalities in women of reproductive age. In this study, we set out to construct a molecular long non-coding RNA (lncRNA)-microRNA (miRNA)-messenger RNA (mRNA) network according to the competitive endogenous RNA (ceRNA) theory and obtain insights into the related biological characteristics and pathways. Methods We downloaded two gene expression profile datasets of mural granulosa cells (MGCs) of women with PCOS and healthy women without PCOS (GSE84376 and GSE106724) from Gene Expression Omnibus (GEO) DataSets. Using GEO2R, we identified the mRNAs and non-coding RNAs with differential expression. The DIANA-microT-CDS algorithm was applied to predict the genes targeted by the differentially expressed miRNAs. The lncRNA-miRNA interactions were predicted using DIANA-LncBase v2. Then, we constructed the lncRNA-miRNA-mRNA network. The Database for Annotation, Visualization and Integrated Discovery (DAVID) was employed to identify the functions and enriched pathways of the genes. Subsequently, STRING was used to construct the protein-protein interaction (PPI) network. cytoHubba in Cytoscape was used to rank the hub genes, and finally, PPI modules were screened with Cytoscape MCODE. Results There were 462 mRNAs, 2,464 lncRNAs, and 55 miRNAs which showed differential expression between the MGCs of patients with PCOS and those of healthy controls. Based on the PPI analysis, differentially expressed genes (DEGs) were significantly enriched in retinol metabolism, drug metabolism—cytochrome P450, malaria, the Hippo signaling pathway, and glycine, serine, and threonine metabolism. The ceRNA network contained 71 lncRNA nodes, 14 miRNA nodes, and 69 mRNA nodes, as well as 167 edges. We identified some novel genes and non-coding RNAs that might be involved in PCOS, including CD163, MRC1, VSIG4, CCL2, CCR2, SPP1, hsa-miR-3135b, hsa-miR-4649-3p, hsa-miR-1231, hsa-miR-3609, and hsa-miR-4433b-3p. Conclusions This study identified a novel lncRNA-miRNA-mRNA network based on the ceRNA mechanism in PCOS. Some novel genes and non-coding RNAs that may be involved in the occurrence and development of PCOS were excavated, including CD163, MRC1, VSIG4, CCL2, CCR2, SPP1, hsa-miR-3135b, hsa-miR-4649-3p, hsa-miR-1231, hsa-miR-3609, and hsa-miR-4433b-3p. However, our findings need to be validated by in vivo and in vitro experiments.

[目的]研究干扰雄性小鼠睾丸Cloc k基因后,对小鼠胚胎着床及后期胎鼠发育的影响,并从甲基化的角度探讨引起胎鼠发育异常的原因.[方法]将成年雄性小鼠分为干扰组和对照组,分别于睾丸中注射干扰质粒和阴性质粒,与经过促排处理的雌鼠合笼,研究比较两组胚胎着床情况,观察两组胎鼠的外形及体重等指标的差异;观察胎鼠组织结构,比较两组是否存在差异;分析胎鼠总甲基化水平,同时测定甲基转移酶(DNMTs)水平,比较两组是否存在差异.[结果]:①与对照组比较,干扰组胚胎着床数下降(14.5 vs.20.33),胎儿体重减轻,死胎和异常胎数增多(P<0.05).②干扰组胎鼠的心脏和脊柱发育分节出现异常.③11.5～16.5 dpc阶段胎鼠总甲基化水平,干扰组较对照组显著降低(8.59 vs.18.25);干扰组甲基转移酶DNMT3B-4的相对IOD值低于对照组(0.124 vs.0.141),有统计学意义.[结论]干扰雄性小鼠睾丸Clock基因,可导致胚胎着床数下降,胎鼠发育异常增多,体重减轻.甲基转移酶DNMT3B-4水平下调及总甲基化水平下降,可能是影响胎鼠发育异常的原因.本研究表明Clock基因与胎鼠发育密切相关,初步探讨引起不育的机制,为揭示该类疾病发病机制提供依据.

目的:筛选结肠癌(Colorectal cancer,CRC)癌组织与正常癌旁组织之间差异表达基因(Differentially Expressed Genes,DEGs),分析结肠癌的发病机制和可能的生物标志物.方法:在基因表达综合数据库(Gene Expression Omnibus,GEO)中以"colon cancer"作为搜索词,检索关于CRC的基因表达芯片数据.采用R语言对获取的芯片数据进行基因的差异表达分析,对所得DEGs进行基因本(Gene Ontology,GO)和京都基因与基因组百科全书(Kyoto Encyclopedia of Genes and Genomes,KEGG)信号通路的富集分析,并对DEGs的编码蛋白进行蛋白互作分析,筛选其中的关键基因(hub gene).结果:在芯片GSE44861中共筛选出241个DEGs,包括74个上调DEGs和167个下调DEGs.GO和KEGG分析显示,DEGs分别在8条生物过程(Biological Process,BP)、5条细胞组成(Cellular Component,CC)、13条分子功能(Molecular Function,MF)注释和2条KEGG通路中富集.所有互作蛋白中筛选出10个hub基因,除P2RY14外,其余主要分布于MF注释的"CXCR趋化因子受体结合"、"趋化因子受体结合"、"趋化因子活性"、"受体配体活性"、"信号受体激活剂活性"、"激素活性",以及BP注释的"抗菌肽介导的抗菌体液免疫应答"、"抗菌体液反应"、"体液免疫反应".结论:CRC的发生可能与CXCR趋化因子受体结合以及肠道感染和免疫应答密切相关,P2RY14有可能成为CRC的筛查、诊断、预后的分子生物标志物.

A telemetry system based on Bluetooth Low Energy (BLE) was constructed to simultaneously collect locomotor activity and physiological signals of small animal cohorts for circadian rhythm experiments; it consists of miniature transmitters and mobile phone with customized App. The continuous sampling signals obtained from the 3-axis acceleration and temperature sensors in the transmitters are sent to the mobile phone in real-time through Internet of Things (IoT) for temporary storage and then imported into the computer for summary and rhythm analysis by the general open-source mathematical software. Unlike expensive and complicated commercial telemetry systems with industrial wireless standards, no special data receivers and software are needed. In our validation experiment, six rats were divided into two groups under natural dark and light-dark cycles. For two consecutive weeks, the transmitter mounted on the head of the rat-recorded locomotor activity, skin temperature, and ambient temperature of each rat at a frequency of 6 Hz. After processing with Local Weighted Regression Scatter Smoothing (LOWESS) and Fast Fourier Transform (FFT) filtering, single cosinor and multi-components cosinor were then used to assess and characterize the circadian rhythm. The results showed that the rhythm values of the two groups of rats coincided with the corresponding light-dark cycle, and that the system was robust to data loss and error from BLE communication failures. Therefore, the proposed system provides a light-weight framework for long-term circadian rhythm monitoring in free-moving rodents to further simplify and promote experimental chronobiology animal studies.

Glioma-associated microglial cells, a key component of the tumor microenvironment, play an important role in glioma progression. In this study, the mouse glioma cell line GL261 and the mouse microglia cell line BV2 were chosen. First, circadian gene expression in glioma cells co-cultured with either M1 or M2 microglia was assessed and the exosomes of M2-polarized and unpolarized BV-2 microglia were extracted. Subsequently, we labeled the exosomes with PKH67 and treated GL261 cells with them to investigate the exosome distribution. GL261 cell phenotypes and related protein expression were used to explore the role of M2 microglial exosomes in gliomas. Then a specific miR-7239-3p inhibitor was added to verify miR-7239-3p functions. Finally, the mouse subcutaneous tumorigenic model was used to verify the tumorigenic effect of M2 microglial exosomes in vivo. Our results showed that in gliomas co-cultured with M2 microglia, the expression of the BMAL1 protein was decreased (P < 0.01), while the expression of the CLOCK protein was increased (P < 0.05); opposite results were obtained in gliomas co-cultured with M1 microglia. After treatment with M2 microglial exosomes, the apoptosis of GL261 cells decreased (P < 0.001), while the viability, proliferation, and migration of GL261 cells increased. Increased expression of N-cadherin and Vimentin, and decreased E-cadherin expression occurred upon treatment with M2 microglial exosomes. Addition of an miR-7239-3p inhibitor to M2 microglial exosomes reversed these results. In summary, we found that miR-7239-3p in the glioma microenvironment is recruited to glioma cells by exosomes and inhibits Bmal1 expression. M2 microglial exosomes promote the proliferation and migration of gliomas by regulating tumor-related protein expression and reducing apoptosis.

Emerging evidence has demonstrated that long noncoding RNAs (lncRNAs) play critical roles in the epigenetic and transcriptional regulation of mammalian circadian systems. Circadian rhythmicity regulates many aspects of our immune system, and perturbation of the circadian clock can augment the inflammatory response. However, knowledge of the precise functions of lncRNAs in the regulation of immune functions within the circadian system is relatively limited. In this study, differentially expressed lncRNAs induced by Clock knockdown were screened via mRNA/lncRNA microarray and bioinformatic prediction analysis. We identified a Clock-regulated lncRNA, AK028245, which was correlated with the activation of the immune response. The expression levels of AK028245 were decreased in the spleen of immunosuppressed mice and elevated in immune-activated mice treated with lipopolysaccharide (LPS). Further, Clock knockdown decreased the expression of OTUD7B and A20, 2 early immune response factors acting on the NF-κB signaling pathway. Interestingly, inhibition of AK028245 increased their expression, mitigating the effects of Clock knockdown. In addition, inhibition of AK028245 downregulated the expression of tumor necrosis factor-α and interleukin-6 in the late stages of LPS stimulation and the expression of interferon-γ and Cxcl12 in the peak stages. We conclude that this newly identified lncRNA plays a role in the crosstalk between Clock and immune response regulators, likely resulting in a proinflammatory response targeting OTUD7B and A20. The lncRNA AK028245 has revealed a new mechanism of the immune response and provided new targets for the treatment of immune disorders.

Our previous study indicated that Clock gene could affect the thrombotic potential. In this present study, we examined the differential expression of proteins in Clock knockdown mice's plasma, including alpha 1-antitrypsin as a potential target. The proteins were examined in AML12 cells with Clock gene being knocked down to confirm the differential expressions. RNAs of those cells were extracted every 3 h in 24 h. The transcriptional levels of alpha 1-antitrypsin (Serpina1a) and fibronectin (Fn1) were analyzed with the least-squares fit of a 24-h cosine function by single cosinor method, but no circadian rhythm was determined in neither of these genes. The expressions of alpha 1-antitrypsin and fibronectin in Clock knockdown cells were found upregulated in both transcriptional and translational levels. Then, we applied ELISA assay to detect the concentration of activated protein C in Clock knockdown plasma and found a risen concentration. Fibronectin was reported to play a role in inhibiting the platelet aggregation, while activated protein C was demonstrated to inhibit PAI-1. All these results indicated that downregulation of the Clock gene in the circulatory system might have effects on PAI-1 by upregulating alpha 1-antitrypsin, and might play some roles in platelet aggregation by increased fibronectin.

This study was conducted to identify genes that are differentially expressed in paracancerous tissue and to determine the potential predictive value of selected gene panel. Gene transcriptome data of bladder tissue was downloaded from UCSC Xena browser and NCBI GEO repository, including GTEx (the Genotype-Tissue Expression project) data, TCGA (The Cancer Genome Atlas) data, and GEO (Gene Expression Omnibus) data. Differentially Expressed Genes (DEGs) analysis was performed to identify tumor-DEGs candidate genes, using the intersection of tumor-paracancerous DEGs genes and paracancerous-normal DEGs genes. The survival-related genes were screened by Kaplan–Meier (KM) survival analysis and univariable Cox regression with the cutoff criteria of KM < 0.05 and cox p -value < 0.05. The risk model was developed using Lasso regression. The clinical data were analyzed by univariate and multivariate Cox regression analysis. Gene Ontology (GO) and KEGG enrichment analysis were performed in the DEGs genes between the high-risk and low-risk subgroups. We identified six survival-related genes, EMP1, TPM1, NRP2, FGFR1, CAVIN1, and LATS2, found in the DEG analyses of both, tumor-paracancerous and paracancerous-normal differentially expressed data sets. Then, the patients were classified into two clusters, which can be distinguished by specific clinical characteristics. A three-gene risk prediction model (EMP1, FGFR1, and CAVIN1) was constructed in patients within cluster 1. The model was applied to categorize cluster 1 patients into high-risk and low-risk subgroups. The prognostic risk score was considered as an independent prognostic factor. The six identified survival-related genes can be used in molecular characterization of a specific subtype of bladder cancer. This subtype had distinct clinical features of T (topography), N (lymph node), stage, grade, and survival status, compared to the other subtype of bladder cancer. Among the six identified survival-related genes, three-genes, EMP1, FGFR1, and CAVIN1, were identified as potential independent prognostic markers for the specific bladder cancer subtype with clinical features described.

Circadian rhythm has been involved in the regulation of many physiological activities. Autophagy is the metabolic process that transports substances in the cytoplasm to the lysosomes for degradation, and involved in the process of many diseases, including cancer. Studies have shown that autophagy activity has a circadian rhythm feature. As the core gene of the circadian rhythm system, clock has participated in the occurrence and development of cancer. The expression of clock whether regulates tumor development by autophagy has not been illustrated at present. In this study, we established a stableclock-knockdown strain of mouse breast cancer cell 4T1 to explore the changes in autophagy activity. The results showed that autophagy-related proteins ATG9A, ATG7, LC3B and Beclin1 (ATG6) were down-regulated withclock-knockdown, and the p62, the key factors of autophagy, was up-regulated. Moreover,clock-knockdown caused significant up-regulation of p65, and inhibition of I kappa B alpha and p-AKT?which were the core factors of several signaling pathways involved in autophagy. Serum rhythm-inducing experiments behaved that the expression of LC3B and Beclin1 had rhythmic characteristics?differed from LC3A. The regulative geneclockwas initially established to regulate the autophagy-related genes, and the suggested direction of its regulatory pathways.

Circadian rhythms help organisms adapt to changes of external environment by regulating energy metabolism and remaining the balance of homeostasis. Numerous researches have proved that the physiological function of liver was precisely controlled by circadian rhythms. Clock , one of core circadian genes, has been demonstrated to regulate the oxidative phosphorylation process of mitochondrial, which provides energy for living cells and acts as one of the hub for apoptosis. However, whether Clock gene regulates mitochondrial apoptosis pathways in liver cells remains less explored. In the present study, we used lentiviral vector to establish a stable AML12 cell lines which were capable of expressing specific shRNA to interfere the expression of Clock gene and investigated the effect of Clock on mitochondrial apoptosis pathways. Herein, we found that the interference of Clock gene could significantly suppress mitochondrial apoptosis pathways by stabilizing mitochondrial membrane potential and inhibiting mitochondria out membrane permeablization, which might be a result of lower expression of BAD and BIM proteins. Moreover, the interference of Clock gene could downregulate the expression of mitochondrial apoptosis factors, i.e. AIF, CYCS, APAF-1 and SMAC, which will suppress the formation of apoptosome and the process of DNA degradation to further inhibit apoptosis process. This work provides an insight on the important role of Clock gene participating in mitochondrial apoptosis pathways of hepatocytes and unveils a probable pathogenesis of how circadian rhythm regulates liver diseases.

目的 探讨不明原因复发性自然流产(URSA)患者子宫内膜差异基因筛选及其相关信号通路及生物学过程.方法 在基因表达总览(GEO)数据库中,以"recurrent spontaneous abortion""recurrent pregnancy loss""endometrium""homo sapiens"为关键词,检索URSA患者子宫内膜基因芯片数据集.利用R语言对U RS A患者子宫内膜基因芯片数据进行分析,筛选差异基因,并对不同芯片数据集间差异基因取交集,获得共同差异基因.利用David数据库对共同差异基因的基因本体(GO)及京都基因与基因组百科全书(KEGG)信号通路进行富集分析.利用STRING数据库对这些基因数据集的差异基因编码蛋白的蛋白质与蛋白质相互作用(PPI)网络进行构建,并采用Cytoscape软件(v3.0)进行可视化.结果 ①本研究获得GSE26787和GSE65099共计2个URSA患者子宫内膜基因数据集,对这2个数据集差异基因取交集,最终获得23个共差异基因,其中上调基因为19个,分别为ADM2、SH3D21、FGFR4、SULT2B1、NRG2、SERPINA4、NLRP5、FAM124B、MCOLN3、SLC7A1、PPP1R1B、SQLE、SLC24A4、PLPPR5、EPHB3、SMIM24、ZNF589、SOX7、EDN3;下调基因为4个,分别为PRKCB、A NG、IRX5、A TG9B.②共同差异基因GO富集分析获得BP、CC和M F共计3个部分结果.共同差异基因在BP中,主要涉及细胞Ca2+稳态、血管生成及细胞内信号传导;在CC中,主要涉及膜的整体组分;在M F中,主要涉及受体结合.③K EGG通路富集分析结果显示,共同差异基因富集于ErbB信号通路(P=0.070,FDR=55.245).涉及该信号通路的共同差异基因为NRG2和PRKCB.结论 基于GEO数据库,采用生物信息学的方法筛选到参与URSA发生、发展的部分差异基因,并为进一步研究提供数据支撑.