Coordination of zygotic genome activation entry and exit by H3K4me3 and H3K27me3 in porcine early embryos.

Histone modifications are critical epigenetic indicators of chromatin state associated with gene expression. Although the reprogramming patterns of H3K4me3 and H3K27me3 have been elucidated in mouse and human preimplantation embryos, the relationship between these marks and zygotic genome activation (ZGA) remains poorly understood. By ultra-low-input native chromatin immunoprecipitation and sequencing, we profiled global H3K4me3 and H3K27me3 in porcine oocytes and in vitro fertilized (IVF) embryos. We found that promoters of ZGA genes occupied sharp H3K4me3 peaks in oocytes, and these peaks became broader after fertilization, and reshaped into sharp again during ZGA. By simultaneous depletion of H3K4me3 demethylase KDM5B and KDM5C, we determined that broad H3K4me3 domain maintenance impaired ZGA gene expression, suggesting its function to prevent premature ZGA entry. By contrast, broad H3K27me3 domains underwent global removal upon fertilization, followed by a re-establishment for H3K4me3/H3K27me3 bivalency in morulae. We also found that bivalent marks were deposited at promoters of ZGA genes, and inhibiting this deposition was correlated with the activation of ZGA genes. It suggests that promoter bivalency contributes to ZGA exit in porcine embryos. Moreover, we demonstrated that aberrant reprogramming of H3K4me3 and H3K27me3 triggered ZGA dysregulation in somatic cell nuclear transfer (SCNT) embryos, whereas H3K27me3-mediated imprinting did not exist in porcine IVF and SCNT embryos. Our findings highlight two previously unknown epigenetic reprogramming modes coordinated with ZGA in porcine preimplantation embryos. Finally, the similarities observed between porcine and human histone modification dynamics suggest that the porcine embryo may also be a useful model for human embryo research.

MnO2-based dual channel surface plasmon resonance fiber sensor for trace glutathione and refractive index detection.

Glutathione (GSH) plays vital role in human biological systems, so its rapid and sensitive detection is necessary for health condition monitoring. In this work, a simple structure for dual channel GSH and refractive index (RI) detection is proposed. By introducing Au-MnO2 thin film coating on the fiber surface for the first time, GSH solution would lead to the dissolution of MnO2, the change in GSH levels could be monitored over a short period in channel 2. For channel 1, ITO-Ag thin film is applied for RI change detection. After optimization, the GSH detection sensitivity reached about -2.361 nm/mM in the range of 0.005-50 mM, and the RI sensitivity reached 1704.252 nm/RIU in the range of 1.331-1.3895 RIU. Channel 1 could also put into GSH detection in the high concentration scale to enlarge the sensor's range and 0.095 nm/mM of sensitivity is acquired within the range of 50-600 mM. With the presence of MnO2 film, the detection sensitivity increased 25.663 times. Neither channel interferes with the operation of the other. Proposed sensor provides stability, high selectivity and elevation in GSH detection sensitivity, which shows great potential for environmental and biological detection field and their applications.

Highly sensitive surface plasmon resonance temperature sensor based on a hollow core fiber multilayer structure.

A surface plasmon resonance (SPR) temperature sensor based on a hollow core fiber (HCF) is designed in this paper. The sensor is composed of a multi-mode fiber (MMF)-HCF-MMF structure, and the self-made HCF is deposited successively with a thin layer of Au film (50 nm in thickness), gold nanoparticles (10 nm in diameter) and polydimethylsiloxane (PDMS). A series of theoretical and experimental investiagtions are conducted, and the results are as follows: the proposed sensing structure only with Au film can effectively excite the SPR effect, with a sensitivity of (2200 ± 100) nm / RIU in the refractive index (RI) range of 1.3334-1.3811; after adding AuNPS, the sensitivity of the sensor is effectively improved, the sensitivity can be increased to (3100 ± 100) nm / RIU, and after the PDMS coating, temperature sensing can be realized due to its unique temperature-sensitive characteristics, a linear sensitivity of (-2.1 ± 0.1) nm / °C is realized in the temperature range of 25 °C to 100 °C. The sensor has the advantages of simple structure, wide application, large measurement range, high sensitivity, good stability and repeatability. Meanwhile, the internal air hole of HCF leaves a preparation channel for dual-parameter measurement. It has broad application prospect in medical treatment, environmental monitoring and manufacturing industry.

Circ_0000064 promotes high glucose-induced renal tubular epithelial cells injury to facilitate diabetic nephropathy progression through miR-532-3p/ROCK1 axis.

BACKGROUND: Circular RNA (circRNA) has been shown to mediate diabetic nephropathy (DN) development by regulating renal tubular epithelial cells (RTECs) injury. However, the role and mechanism of circ_0000064 in high glucose (HG)-induced RTECs injury have not been fully elucidated. METHODS: Human RTECs (HK-2) were exposed to HG to induce cell injury. Cell oxidative stress was assessed by detecting the levels of oxidative stress-markers. Moreover, cell proliferation and apoptosis were determined by CCK8 assay, EDU assay and flow cytometry. The protein levels of proliferation markers, apoptosis markers and Rho-associated coiled-coil-containing kinase 1 (ROCK1) were measured using western blot analysis. Furthermore, quantitative real-time PCR was performed to assess the expression of circ_0000064, microRNA (miR)-532-3p and ROCK1. The interaction between miR-532-3p and circ_0000064 or ROCK1 was confirmed by dual-luciferase reporter assay and RNA pull-down assay. RESULTS: Our results revealed that HG treatment could promote HK-2 cells oxidative stress, apoptosis, fibrosis, and inhibit proliferation. Circ_0000064 expression was increased in the serum of DN patients and HG-induced HK-2 cells, and silenced circ_0000064 could relieve HG-induced HK-2 cells injury. MiR-532-3p could be sponged by circ_0000064, and its overexpression also alleviated HG-induced HK-2 cells injury. Besides, the regulation of circ_0000064 knockdown on HG-induced HK-2 cells injury could be reversed by miR-532-3p inhibitor. Additionally, ROCK1 was a target of miR-532-3p, and its expression was inhibited by circ_0000064 knockdown. The inhibition effect of circ_0000064 knockdown on HG-induced HK-2 cells injury also could be reversed by overexpressing ROCK1. CONCLUSION: In summary, circ_0000064 knockdown might alleviate HG-induced HK-2 cells injury via regulating the miR-532-3p/ROCK1 axis, which provided a new perspective for DN treatment.

An Adaptive Feature Selection Algorithm for Fuzzy Clustering Image Segmentation Based on Embedded Neighbourhood Information Constraints.

This paper addresses the lack of robustness of feature selection algorithms for fuzzy clustering segmentation with the Gaussian mixture model. Assuming that the neighbourhood pixels and the centre pixels obey the same distribution, a Markov method is introduced to construct the prior probability distribution and achieve the membership degree regularisation constraint for clustering sample points. Then, a noise smoothing factor is introduced to optimise the prior probability constraint. Second, a power index is constructed by combining the classification membership degree and prior probability since the Kullback-Leibler (KL) divergence of the noise smoothing factor is used to supervise the prior probability; this probability is embedded into Fuzzy Superpixels Fuzzy C-means(FSFCM) as a regular factor. This paper proposes a fuzzy clustering image segmentation algorithm based on an adaptive feature selection Gaussian mixture model with neighbourhood information constraints. To verify the segmentation performance and anti-noise robustness of the improved algorithm, the fuzzy C-means clustering algorithm Fuzzy C-means (FCM), FSFCM, Spatially Variant Finite Mixture Model(SVFMM), EGFMM, extended Gaussian mixture model (EGMM), adaptive feature selection robust fuzzy clustering segmentation algorithm (AFSFCM), fast and robust spatially constrained Gaussian mixture model (GMM) for image segmentation (FRSCGMM), and improve method are used to segment grey images containing Gaussian noise, salt-and-pepper noise, multiplicative noise and mixed noise. The peak signal-to-noise ratio (PSNR) and the error rate (MCR) are used as the theoretical basis for assessing the segmentation results. The improved algorithm indicators proposed in this paper are optimised. The improved algorithm yields increases of 0.1272-12.9803 dB, 1.5501-13.4396 dB, 1.9113-11.2613 dB and 1.0233-10.2804 dB over the other methods, and the Misclassification rate(MSR) decreases by 0.32-37.32%, 5.02-41.05%, 0.3-21.79% and 0.9-30.95% compared to that with the other algorithms. It is verified that the segmentation results of the improved algorithm have good regional consistency and strong anti-noise robustness, and they meet the needs of noisy image segmentation.

A Local Neighborhood Robust Fuzzy Clustering Image Segmentation Algorithm Based on an Adaptive Feature Selection Gaussian Mixture Model.

Since the fuzzy local information C-means (FLICM) segmentation algorithm cannot take into account the impact of different features on clustering segmentation results, a local fuzzy clustering segmentation algorithm based on a feature selection Gaussian mixture model was proposed. First, the constraints of the membership degree on the spatial distance were added to the local information function. Second, the feature saliency was introduced into the objective function. By using the Lagrange multiplier method, the optimal expression of the objective function was solved. Neighborhood weighting information was added to the iteration expression of the classification membership degree to obtain a local feature selection based on feature selection. Each of the improved FLICM algorithm, the fuzzy C-means with spatial constraints (FCM_S) algorithm, and the original FLICM algorithm were then used to cluster and segment the interference images of Gaussian noise, salt-and-pepper noise, multiplicative noise, and mixed noise. The performances of the peak signal-to-noise ratio and error rate of the segmentation results were compared with each other. At the same time, the iteration time and number of iterations used to converge the objective function of the algorithm were compared. In summary, the improved algorithm significantly improved the ability of image noise suppression under strong noise interference, improved the efficiency of operation, facilitated remote sensing image capture under strong noise interference, and promoted the development of a robust anti-noise fuzzy clustering algorithm.

PDZRN4 acts as a suppressor of cell proliferation in human liver cancer cell lines.

Recently, some reports show that Ligand of Numb Protein-X 1 (LNX1) could be a suppressor gene in gliomas, while our current research has firstly shown that PDZ domain containing ring finger 4 (PDZRN4), another member of LNX family, could also be a potential suppressor in hepatocellular carcinoma (HCC). PDZRN4, also named LNX4 (Ligand of Numb Protein-X 4), is a member of the LNX family. We recently found that PDZRN4, but not LNX1, was down-regulated in HCC samples, and the role of PDZRN4 in the progression of HCC had not been studied before. To address this question, firstly, we evaluated the expression of PDZRN4 in HCC samples and adjacent non-cancerous tissues. Semi-quantitative polymerase chain reaction showed that PDZRN4 was down-regulated in 24/36 (66.7%) HCC samples separately. In addition, our research shows that PDZRN4 is silenced in all of the 12 HCC cell lines tested. Subsequently, cell-based functional assay exhibited that ectopic expression of PDZRN4 inhibits the proliferation, plate colony formation and anchorage-independent colony formation of HCC cells. Collectively, our results showed that PDZRN4 might be a potential tumour suppressor gene and had anti-proliferative effect on HCC cell proliferation, which would be of great significance to the researches on HCC.

TICRR Overexpression Enhances Disease Aggressiveness and Immune Infiltration of Cutaneous Melanoma.

Objective: To investigate the role of the TopBP1 interacting checkpoint and replication regulator (TICRR) in cutaneous melanoma (CM) as a prognostic biomarker and therapeutic target. Methods: TICRR expression in tumour samples was explored using the TCGA and the GTEx database. The Kaplan-Meier survival curve, nomogram model and risk score curve were established to evaluate the prognostic role of TICRR in CM. Tissue samples of CM patients were obtained to validate the TICRR expression further. Several experiments in vitro were conducted to investigate the effect of TICRR upon CM aggressiveness and to explore underlying mechanisms. Results: TICRR was overexpressed in CM tissue and was correlated with poor prognosis of CM patients. The knockdown of TICRR decreased the proliferation, migration, and invasion of CM cells, whereas overexpression produced the opposite effect. Furthermore, TICRR suppression substantially attenuated the activation of PI3K/AKT/mTOR signalling, while the PI3K/AKT inhibitor LY294002 could partially reverse the aggressiveness-enhancing effect induced by TICRR overexpression. It was further confirmed that TICRR was closely related to immune cell infiltration activities by using immune infiltration and immunofluorescence analysis. Conclusion: TICRR overexpression may enhance CM aggressiveness by activating the PI3K/Akt/mTOR pathway and promoting immune infiltration. TICRR was verified as a potential prognostic biomarker and therapeutic target for CM.

KLF4 facilitates chromatin accessibility remodeling in porcine early embryos.

Chromatin accessibility remodeling driven by pioneer factors is critical for the development of early embryos. Current studies have illustrated several pioneer factors as being important for agricultural animals, but what are the pioneer factors and how the pioneer factors remodel the chromatin accessibility in porcine early embryos is not clear. By employing low-input DNase-seq (liDNase-seq), we profiled the landscapes of chromatin accessibility in porcine early embryos and uncovered a unique chromatin accessibility reprogramming pattern during porcine preimplantation development. Our data revealed that KLF4 played critical roles in remodeling chromatin accessibility in porcine early embryos. Knocking down of KLF4 led to the reduction of chromatin accessibility in early embryos, whereas KLF4 overexpression promoted the chromatin openness in porcine blastocysts. Furthermore, KLF4 deficiency resulted in mitochondrial dysfunction and developmental failure of porcine embryos. In addition, we found that overexpression of KLF4 in blastocysts promoted lipid droplet accumulation, whereas knockdown of KLF4 disrupted this process. Taken together, our study revealed the chromatin accessibility dynamics and identified KLF4 as a key regulator in chromatin accessibility and cellular metabolism during porcine preimplantation embryo development.

GATA2 promotes oxidative stress to aggravate renal ischemia-reperfusion injury by up-regulating Redd1.

Renal ischemia-reperfusion injury (RIRI) is a common pathophysiological process, and it is also an important cause of acute renal failure. Therefore, finding an effective therapeutic target for RIRI is extremely urgent. In our study, we constructed hypoxia-reoxygenation (HR) model in vitro and a renal ischemia-reperfusion (IR) model in vivo. Elevated levels of serum creatinine (Cr), blood urea nitrogen (BUN) tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6) and malondialdehyde (MDA) along with the decreased levels of superoxide dismutase (SOD) and glutathione (GSH) proved that kidney function was damaged after IR, and pathological changes of renal tissues were observed using HE staining and TUNEL staining. The protein of Redd1 expression level was detected to be upregulated after IR by western blot (WB). However, transfection of short hairpin RNA of Redd1 (sh-Redd1) alleviated the HR injury on LLC-PK1 cells, as evidenced by increased cell viability, proliferation and decreased cell apoptosis; additionally, the accumulation of ROS was inhibited. Sh-Redd1 also alleviated IR injury in the mouse model. Subsequently, GATA2 was proved to be upregulated in IR and HR models and was the transcription factor of Redd1. Knockdown of GATA2 efficiently mitigated the oxidative stress induced damages in vivo and in vitro, while these mitigations were reversed by transfection of Redd1 overexpression plasmid. In conclusion, our study clarified the possible underlying mechanism of protecting RIRI.

Establishment and engineering application of viscoelastic-plastic constitutive laws for creep modeling in interbedded rock masses.

In order to study the creep behavior of the surrounding rock of the interbedded rock mass tunnel considering the time-dependent deformation, this paper proposes a viscoelastic-plastic seven-element model considering the stress threshold, and derives and establishes its creep equation under three-dimensional stress state. At the same time, the UMAT (User-defined Material) subroutine of the model is developed based on the ABAQUS software. The rationality of the seven-element model and the effectiveness of the subprogram are verified by rheological test results. Finally, the UMAT subroutine is applied to the numerical simulation of the creep behavior of soft and hard interbedded rock tunnels with different rock inclinations (α). The results show that the different rock inclination angles have different effects on the horizontal displacement of the ground above the tunnel, settlement deformation, and the convergence of the tunnel section. With the increase of the rock inclination (0 ≤ α ≤ 90°), the horizontal displacement of the surface on both sides is antisymmetric. When α is 0°, 45° and 90°, the horizontal displacement on both sides is equivalent. Surface subsidence decreases and then increases slowly. When α is 0° and 45°, the surface subsidence is the largest (12.4 mm) and the smallest (11.1 mm), respectively. The convergence values of the tunnel section change according to different parts of the tunnel. The convergence values of the arch top and arch bottom decrease continuously, and their maximum convergence values are 23.4 mm and 17.3 mm, respectively. The change trend of the arch waist and arch shoulder convergence values is the opposite. When α is 0°, the convergence value of the arch waist is maximum (3.5 mm). When α is 15°, the convergence value of the arch shoulder is the maximum (2.0 mm).

Transcriptomic and Physiological Analyses Reveal the Molecular Mechanism through Which Exogenous Melatonin Increases Drought Stress Tolerance in Chrysanthemum.

Chrysanthemum (Chrysanthemum morifolium (Ramat.) Hemsl.) is an important species in China's flower industry, and drought stress seriously affects the growth, quality, yield, and geographical distribution of this species. Melatonin (MT) plays a key role in regulating plant abiotic stress responses and stress resistance, but the mechanism through which exogenous MT regulates drought resistance in chrysanthemum remains unclear. This study explored the protective effect of MT on chrysanthemum drought tolerance and its key regulatory pathways. Exogenous MT application increased the photosynthetic capacity (Tr increased by 18.07%; Pn increased by 38.46%; and Gs increased by 26.52%) of chrysanthemum and attenuated decreases in its chlorophyll (19.89%) and relative water contents (26.94%). Moreover, MT increased the levels of osmolarity-related compounds such as soluble sugars (43.60%) and soluble protein (9.86%) under drought stress and increased antioxidant enzyme activity (SOD increased by 20.98%; POD increased by 35.04%; and CAT increased by 26.21%). Additionally, MT increased the endogenous MT (597.96%), growth hormone (45.31% and 92.09%), gibberellic acid (75.92% and 3.79%), salicylic acid (33.02%), and cytokinin contents (1400.00%) under drought stress while decreasing the abscisic acid (50.69% and 56.79%), jasmonate contents (62.57% and 28.31%), and ethylene contents (9.28%). RNA-seq analysis revealed 17,389, 1466, and 9359 differentially expressed genes (DEGs) under three treatments (PEG, MT, and MT _ PEG, respectively) compared with the control. Enrichment analyses of the DEGs identified more than 10 GO terms and 34 KEGG pathways. Nitrogen metabolism, sulfur metabolism, and alanine, aspartate, and glutamate metabolism were significantly increased under all three treatments. The DEGs included many transcription factors, such as MYB, WRKY, and NAC proteins. Our results preliminarily classify candidate genes and metabolic pathways with active roles in the interaction between MT and drought stress and advance the understanding of the molecular mechanism of the response to drought stress under MT conditions, thereby providing a theoretical basis for the breeding of drought-resistant chrysanthemum.

Emodin Ameliorates High Glucose-Induced Podocyte Apoptosis via Regulating AMPK/mTOR-Mediated Autophagy Signaling Pathway.

OBJECTIVE: To investigate the effect of emodin on high glucose (HG)-induced podocyte apoptosis and whether the potential anti-apoptotic mechanism of emodin is related to induction of adenosine-monophosphate-activated protein kinase (AMPK)/mammalian target of rapamycin (mTOR)-mediated autophagy in podocytes (MPC5 cells) in vitro. METHODS: MPC5 cells were treated with different concentrations of HG (2.5, 5, 10, 20, 40, 80 and 160 mmol/L), emodin (2, 4, 8 µ mol/L), or HG (40 mmol/L) and emodin (4 µ mol/L) with or without rapamycin (Rap, 100 nmol/L) and compound C (10 µ mol/L). The viability and apoptosis of MPC5 cells were detected using cell counting kit-8 (CCK-8) assay and flow cytometry analysis, respectively. The expression levels of cleaved caspase-3, autophagy marker light chain 3 (LC3) I/II, and AMPK/mTOR signaling pathway-related proteins were determined by Western blot. The changes of morphology and RFP-LC3 fluorescence were observed under microscopy. RESULTS: HG at 20, 40, 80 and 160 mmol/L dose-dependently induced cell apoptosis in MPC5 cells, whereas emodin (4 µ mol/L) significantly ameliorated HG-induced cell apoptosis and caspase-3 cleavage (P<0.01). Emodin (4 µ mol/L) significantly increased LC3-II protein expression levels and induced RFP-LC3-containing punctate structures in MPC5 cells (P<0.01). Furthermore, the protective effects of emodin were mimicked by rapamycin (100 nmol/L). Moreover, emodin increased the phosphorylation of AMPK and suppressed the phosphorylation of mTOR. The AMPK inhibitor compound C (10 µ mol/L) reversed emodin-induced autophagy activation. CONCLUSION: Emodin ameliorated HG-induced apoptosis of MPC5 cells in vitro that involved induction of autophagy through the AMPK/mTOR signaling pathway, which might provide a potential therapeutic option for diabetic nephropathy.

Neural grafts containing exosomes derived from Schwann cell-like cells promote peripheral nerve regeneration in rats.

Background: Schwann cell-like cells (SCLCs), differentiated from mesenchymal stem cells, have shown promising outcomes in the treatment of peripheral nerve injuries in preclinical studies. However, certain clinical obstacles limit their application. Hence, the primary aim of this study was to investigate the role of exosomes derived from SCLCs (SCLCs-exo) in peripheral nerve regeneration. Methods: SCLCs were differentiated from human amniotic mesenchymal stem cells (hAMSCs) in vitro and validated by immunofluorescence, real-time quantitative PCR and western blot analysis. Exosomes derived from hAMSCs (hAMSCs-exo) and SCLCs were isolated by ultracentrifugation and validated by nanoparticle tracking analysis, WB analysis and electron microscopy. A prefabricated nerve graft was used to deliver hAMSCs-exo or SCLCs-exo in an injured sciatic nerve rat model. The effects of hAMSCs-exo or SCLCs-exo on rat peripheral nerve injury (PNI) regeneration were determined based on the recovery of neurological function and histomorphometric variation. The effects of hAMSCs-exo or SCLCs-exo on Schwann cells were also determined via cell proliferation and migration assessment. Results: SCLCs significantly expressed the Schwann cell markers glial fibrillary acidic protein and S100. Compared to hAMSCs-exo, SCLCs-exo significantly enhanced motor function recovery, attenuated gastrocnemius muscle atrophy and facilitated axonal regrowth, myelin formation and angiogenesis in the rat model. Furthermore, hAMSCs-exo and SCLCs-exo were efficiently absorbed by Schwann cells. However, compared to hAMSCs-exo, SCLCs-exo significantly promoted the proliferation and migration of Schwann cells. SCLCs-exo also significantly upregulated the expression of a glial cell-derived neurotrophic factor, myelin positive regulators (SRY-box transcription factor 10, early growth response protein 2 and organic cation/carnitine transporter 6) and myelin proteins (myelin basic protein and myelin protein zero) in Schwann cells. Conclusions: These findings suggest that SCLCs-exo can more efficiently promote PNI regeneration than hAMSCs-exo and are a potentially novel therapeutic approach for treating PNI.

Selective autophagic degradation of ACLY (ATP citrate lyase) maintains citrate homeostasis and promotes oocyte maturation.

Macroautophagy/autophagy is a cellular and energy homeostatic mechanism that contributes to maintain the number of primordial follicles, germ cell survival, and anti-ovarian aging. However, it remains unknown whether autophagy in granulosa cells affects oocyte maturation. Here, we show a clear tendency of reduced autophagy level in human granulosa cells from women of advanced maternal age, implying a potential negative correlation between autophagy levels and oocyte quality. We therefore established a co-culture system and show that either pharmacological inhibition or genetic ablation of autophagy in granulosa cells negatively affect oocyte quality and fertilization ability. Moreover, our metabolomics analysis indicates that the adverse impact of autophagy impairment on oocyte quality is mediated by downregulated citrate levels, while exogenous supplementation of citrate can significantly restore the oocyte maturation. Mechanistically, we found that ACLY (ATP citrate lyase), which is a crucial enzyme catalyzing the cleavage of citrate, was preferentially associated with K63-linked ubiquitin chains and recognized by the autophagy receptor protein SQSTM1/p62 for selective autophagic degradation. In human follicles, the autophagy level in granulosa cells was downregulated with maternal aging, accompanied by decreased citrate in the follicular fluid, implying a potential correlation between citrate metabolism and oocyte quality. We also show that elevated citrate levels in porcine follicular fluid promote oocyte maturation. Collectively, our data reveal that autophagy in granulosa cells is a beneficial mechanism to maintain a certain degree of citrate by selectively targeting ACLY during oocyte maturation.Abbreviations: 3-MA: 3-methyladenine; ACLY: ATP citrate lyase; AMA: advanced maternal age; CG: cortical granule; CHX: cycloheximide; CQ: chloroquine; CS: citrate synthase; COCs: cumulus-oocyte-complexes; GCM: granulosa cell monolayer; GV: germinal vesicle; MII: metaphase II stage of meiosis; PB1: first polar body; ROS: reactive oxygen species; shRNA: small hairpin RNA; SQSTM1/p62: sequestosome 1; TCA: tricarboxylic acid; TOMM20/TOM20: translocase of outer mitochondrial membrane 20; UBA: ubiquitin-associated domain; Ub: ubiquitin; WT: wild-type.

DNMT3a negatively regulates PTEN to activate the PI3K/AKT pathway to aggravate renal fibrosis.

BACKGROUND: Renal fibrosis has become one of the major diseases threatening global public health and harming human life and health. PTEN methylation plays an important role in fibrotic diseases of many organs. However, the relationship between PTEN methylation and renal fibrosis is still elusive. METHODS: In the present study, we established a unilateral ureteral obstruction (UUO) mouse model in vivo and a transforming growth factor ß1 (TGF-ß1)-stimulated renal tubular epithelial cell (HK-2) model in vitro. The degree of renal interstitial fibrosis was detected by haematoxylin-eosin (HE) staining and Masson's trichrome staining. Western blot (WB), qRT-PCR, immunohistochemistry (IHC) and methylation-specific PCR (MSP) analyses were used to determine the mechanism by which PTEN methylation regulates renal fibrosis. The α-SMA fibrosis marker was detected by immunofluorescence (IF). Additionally, the relationship of PTEN and DNMT3a in UUO was determined by ChIP-qRT-PCR. RESULTS: Our results showed that the promoter region of PTEN was methylated in UUO. Compared to the sham group, the expression of PTEN was significantly reduced in the UUO group. However, the demethylation reagent significantly inhibited epithelial-mesenchymal transition (EMT), which showed increased expression of E-cadherin and decreased expression of α-SMA and fibronectin. Moreover, treatment of HK-2 cells with 5-aza-dc reversed the activation of the TGF-ß1-induced PI3K/AKT signalling pathway, which inhibited renal fibrosis. WB analysis demonstrated that TGF-ß1 inhibited the PTEN protein expression level and DNMT3a knockdown reversed the inhibitory effect of TGF-ß1 on PTEN expression. Furthermore, ChIP-qRT-PCR showed that DNMT3a interacted with PTEN. Finally, we found that DNMT3a negatively regulated PTEN to activate the PI3K/AKT signalling pathway and aggravate renal fibrosis in vitro and in vivo. CONCLUSION: In summary, these results indicated that renal fibrosis is related to the downregulation of PTEN. Additionally, DNMT3a negatively regulates PTEN to activate the PI3K/AKT signalling pathway and induce EMT in renal tubular epithelial cells, thereby aggravating renal fibrosis.

Hybrid Event-Triggered and Impulsive Control Strategy for Multiagent Systems With Switching Topologies.

This article investigates the hybrid event-triggered and impulsive consensus problems for leaderless and leader-following multiagent systems (MASs) with switching topologies. Based on the state information of neighboring agents at event-triggered moments and impulsive instants, a hybrid event-triggered and impulsive control strategy (HETICS) is designed to reduce the communication frequency between neighboring agents and to ensure consensus of leaderless and leader-following MASs. By utilizing the Lyapunov direct method, some consensus criteria are obtained for leaderless and leader-following MASs with switching topologies. It is shown that the HETICS excludes the Zeno behavior. Several numerical examples and simulations are given to illustrate the effectiveness of the proposed consensus strategy and a comparison with previous consensus control methods is given.

State-based event-triggered consensus strategy for Takagi-Sugeno fuzzy fractional-order multiagent systems with switching topologies.

This article addresses the event-triggered consensus problem for Takagi-Sugeno fuzzy fractional-order multiagent systems with switching topologies. First, to effectively avoid the frequent communication among agents, a state-based event-triggered consensus strategy is designed, which uses the local information from neighboring agents at sampling moments. Then, several sufficient conditions, which rely on the fractional derivative number and time delay information, are presented to guarantee the consensus of fractional-order multiagent systems based on Takagi-Sugeno fuzzy model. Moreover, Zeno behaviors are precluded by proving that the interval length of the two consecutive event-triggering moments for each agent is greater than a positive constant. Finally, some numerical examples are presented, which not only demonstrated the rationality of our proposed consensus protocol but also shown that the presented consensus method based on the designed event-triggered control protocol has the advantage for avoiding communication congestion compared to the existing results.

First-Principles Study of Electronic Properties of Substitutionally Doped Monolayer SnP3.

SnP3 has a great prospect in electronic and thermoelectric device applications due to its moderate band gap, high carrier mobility, absorption coefficients, and dynamical and chemical stability. Doping in two-dimensional semiconductors is likely to display various anomalous behaviors when compared to doping in bulk semiconductors due to the significant electron confinement effect. By introducing foreign atoms from group III to VI, we can successfully modify the electronic properties of two-dimensional SnP3. The interaction mechanism between the dopants and atoms nearby is also different from the type of doped atom. Both Sn7BP24 and Sn7NP24 systems are indirect bandgap semiconductors, while the Sn7AlP24, Sn7GaP24, Sn7PP24, and Sn7AsP24 systems are metallic due to the contribution of doped atoms intersecting the Fermi level. For all substitutionally doped 2D SnP3 systems considered here, all metallic systems are nonmagnetic states. In addition, monolayer Sn7XP24 and Sn8P23Y may have long-range and local magnetic moments, respectively, because of the degree of hybridization between the dopant and its adjacent atoms. The results complement theoretical knowledge and reveal prospective applications of SnP3-based electrical nanodevices for the future.

Hierarchical Accumulation of Histone Variant H2A.Z Regulates Transcriptional States and Histone Modifications in Early Mammalian Embryos.

Early embryos undergo extensive epigenetic reprogramming to achieve gamete-to-embryo transition, which involves the loading and removal of histone variant H2A.Z on chromatin. However, how does H2A.Z regulate gene expression and histone modifications during preimplantation development remains unrevealed. Here, by using ultra-low-input native chromatin immunoprecipitation and sequencing, the genome-wide distribution of H2A.Z is delineated in mouse oocytes and early embryos. These landscapes indicate that paternal H2A.Z is removed upon fertilization, followed by unbiased accumulation on parental genomes during zygotic genome activation (ZGA). Remarkably, H2A.Z exhibits hierarchical accumulation as different peak types at promoters: promoters with double H2A.Z peaks are colocalized with H3K4me3 and indicate transcriptional activation; promoters with a single H2A.Z peak are more likely to occupy bivalent marks (H3K4me3+H3K27me3) and indicate development gene suppression; promoters with no H2A.Z accumulation exhibit persisting gene silencing in early embryos. Moreover, H2A.Z depletion changes the enrichment of histone modifications and RNA polymerase II binding at promoters, resulting in abnormal gene expression and developmental arrest during lineage commitment. Furthermore, similar transcription and accumulation patterns between mouse and porcine embryos indicate that a dual role of H2A.Z in regulating the epigenome required for proper gene expression is conserved during mammalian preimplantation development.