Meiotic transcriptional reprogramming mediated by cell-cell communications in humans and mice revealed by scATACseq and scRNA-seq

Meiosis is a highly complex process significantly influenced by transcriptional regulation.However,studies on the mechanisms that govern transcriptomic changes during meiosis,especially in prophase I,are limited.Here,we performed single-cell ATAC-seq of human testis tissues and observed reprogramming during the transition from zygotene to pachytene spermatocytes.This event,conserved in mice,involved the deactivation of genes associated with meiosis after reprogramming and the activation of those related to spermatogenesis before their functional onset.Furthermore,we identified 282 transcriptional regulators(TRs)that underwent activation or deactivation subsequent to this process.Evidence suggested that physical contact signals from Sertoli cells may regulate these TRs in spermatocytes,while secreted ENHO signals may alter metabolic patterns in these cells.Our results further indicated that defective transcriptional reprogramming may be associated with non-obstructive azoospermia(NOA).This study revealed the importance of both physical contact and secreted signals between Sertoli cells and germ cells in meiotic progression.

Prediction of cell-cell communication patierns of dorsal root ganglion cells:single-cell RNA sequencing data analysis

Dorsal root ganglion neurons transmit peripheral somatic information to the central nervous system,and dorsal root ganglion neuron excitability affects pain perception.Dorsal root ganglion stimulation is a new approach for managing pain sensation.Knowledge of the cell-cell communication among dorsal root ganglion cells may help in the development of new pain and itch management strategies.Here,we used the single-cell RNA-sequencing(scRNA-seq)database to investigate intercellular communication networks among dorsal root ganglion cells.We collected scRNA-seq data from six samples from three studies,yielding data on a total of 17,766 cells.Based on genetic profiles,we identified satellite glial cells,Schwann cells,neurons,vascular endothelial cells,immune cells,fibroblasts,and vascular smooth muscle cells.Further analysis revealed that eight types of dorsal root ganglion neurons mediated proprioceptive,itch,touch,mechanical,heat,and cold sensations.Moreover,we predicted several distinct forms of intercellular communication among dorsal root ganglion cells,including cell-cell contact,secreted signals,extracellular matrix,and neurotransmitter-mediated signals.The data mining predicted that Mrgpra3-positive neurons robustly express the genes encoding the adenosine Adora2b(A2B)receptor and glial cell line-derived neurotrophic factor family receptor alpha 1(GFRα-1).Our immunohistochemistry results confirmed the coexpression of the A2B receptor and GFRα-1.Intrathecal injection of the A2B receptor antagonist PSB-603 effectively prevented histamine-induced scratching behaviour in a dose-dependent manner.Our results demonstrate the involvement of the A2B receptor in the modulation of itch sensation.Furthermore,our findings provide insight into dorsal root ganglion cell-cell communication patterns and mechanisms.Our results should contribute to the development of new strategies for the regulation of dorsal root ganglion excitability.

Identification of an immune-related gene signature for predicting prognosis and immunotherapy efficacy in liver cancer via cell-cell communication

BACKGROUND Liver cancer is one of the deadliest malignant tumors worldwide.Immunotherapy has provided hope to patients with advanced liver cancer,but only a small fraction of patients benefit from this treatment due to individual differences.Identifying immune-related gene signatures in liver cancer patients not only aids physicians in cancer diagnosis but also offers personalized treatment strategies,thereby improving patient survival rates.Although several methods have been developed to predict the prognosis and immunotherapeutic efficacy in patients with liver cancer,the impact of cell-cell interactions in the tumor microenvir-onment has not been adequately considered.AIM To identify immune-related gene signals for predicting liver cancer prognosis and immunotherapy efficacy.METHODS Cell grouping and cell-cell communication analysis were performed on single-cell RNA-sequencing data to identify highly active cell groups in immune-related pathways.Highly active immune cells were identified by intersecting the highly active cell groups with B cells and T cells.The significantly differentially expressed genes between highly active immune cells and other cells were subsequently selected as features,and a least absolute shrinkage and selection operator(LASSO)regression model was constructed to screen for diagnostic-related features.Fourteen genes that were selected more than 5 times in 10 LASSO regression experiments were included in a multivariable Cox regression model.Finally,3 genes(stathmin 1,cofilin 1,and C-C chemokine ligand 5)significantly associated with survival were identified and used to construct an immune-related gene signature.RESULTS The immune-related gene signature composed of stathmin 1,cofilin 1,and C-C chemokine ligand 5 was identified through cell-cell communication.The effectiveness of the identified gene signature was validated based on experi-mental results of predictive immunotherapy response,tumor mutation burden analysis,immune cell infiltration analysis,survival analysis,and expression analysis.CONCLUSION The findings suggest that the identified gene signature may contribute to a deeper understanding of the activity patterns of immune cells in the liver tumor microenvironment,providing insights for personalized treatment strategies.

Source-Load Coordinated Optimal Scheduling Considering the High Energy Load of Electrofused Magnesium and Wind Power Uncertainty

In fossil energy pollution is serious and the“double carbon”goal is being promoted,as a symbol of fresh energy in the electrical system,solar and wind power have an increasing installed capacity,only conventional units obviously can not solve the new energy as the main body of the scheduling problem.To enhance the systemscheduling ability,based on the participation of thermal power units,incorporate the high energy-carrying load of electro-melting magnesiuminto the regulation object,and consider the effects on the wind unpredictability of the power.Firstly,the operating characteristics of high energy load and wind power are analyzed,and the principle of the participation of electrofusedmagnesiumhigh energy-carrying loads in the elimination of obstructedwind power is studied.Second,a two-layer optimization model is suggested,with the objective function being the largest amount of wind power consumed and the lowest possible cost of system operation.In the upper model,the high energy-carrying load regulates the blocked wind power,and in the lower model,the second-order cone approximation algorithm is used to solve the optimizationmodelwithwind power uncertainty,so that a two-layer optimizationmodel that takes into account the regulation of the high energy-carrying load of the electrofused magnesium and the uncertainty of the wind power is established.Finally,the model is solved using Gurobi,and the results of the simulation demonstrate that the suggested model may successfully lower wind abandonment,lower system operation costs,increase the accuracy of day-ahead scheduling,and lower the final product error of the thermal electricity unit.

Regeneration of Diploid Intergeneric Somatic Hybrid Plants Between Microcitrus and Citrus via Electrofusion

Leaf-derived protoplasts of Rough lemon (Citrus jambhiri Lush, 2n = 2x = 18) were electrofused with embryogenic suspension protoplasts of its relative, Microcitrus papuana Swingle (2n = 2x = 18), with an intention of creating novel germplasm. Six plants were regenerated following protoplasts fusion. Cytological examination demonstrated that they were diploids with 18 chromosomes (2n = 2x = 18). RAPD (random amplified polymorphic DNA) analyses with six arbitrary 10-mer primers showed that the regenerated plants had identical band patterns to those of Rough lemon for primers OPA-07, OPAN-07, OPE-05 and OPA-08, Whereas for the other two primers, OPA-04 and OPS-13, bands specific to M. papuana could be detected in the regenerated plants. Cytological and RAPD analysis revealed that the regenerated plants were diploid somatic hybrids between M. papuana and Rough lemon. The putative hybrids were morphologically similar to Rough lemon. This is the first report on production of diploid somatic hybrid plants between citrus with its related genus via symmetric fusion.

Study on the Basic Mechanical Behaviors of High Density Polyethylene Electrofusion Welded Joints at Different Temperature

The basic mechanical behaviors of high density polyethylene electrofusion welded joint at different temperatures were studied by using differently designed specimens in this paper. The results show that the strength of weld bonding plane is higher than that of the pipe and socket materials at room temperature. In order to get the shear strength of electrofusion welded joint, the effective bond lengths were reduced by cutting artificial groove through the socket. The effective bonding length of welded joint to get the shear strength is decreased with decreasing testing temperature. The shear strength and the sensibility to sharp notch of HDPE material increased with decreasing temperature.

Improvement of chromium biosorption through protoplast electrofusion between Candida tropicalis and Candida lipolytica

Protoplasts from Candida tropicalis and Candida lipolytica were fused under an optimized electrofusion （electrical pulse strength 6 kV/cm, pulse duration time 40μs and pulse times 5） and then regenerated on YEPD media for achieving new genotypes with higher chromium loading capacity. A target fusant RHJ-004 was screened out by its chromium resistance and chromium-sorbing capacity tests for further research. The comparative study of applicability shows that the fusant has better performance than its parent strains in respect of solution pH, biomass concentration and chromium loading capacity. Especially for treating low concentration Cr（VI） （〈20 mg/L）, above 80% chromium is sequestered from the aqueous phase at pH 1-9. Atomic force microscopy （AFM） visualizes the distribution of chromium on the binding sites of the cells, suggesting that the altered surface structure and intracellular constitutes of the fusant associate with its increased biosorption capacity. The rapid biosorption processes of chromium foUow the Langmuir model well.

Dynamic interplay between adhesion surfaces in carcinomas:Cell-cell and cell-matrix crosstalk

Cell-cell and cell-matrix signaling and communication between adhesion sites involve mechanisms which are required for cellular functions during normal development and homeostasis; however these cellular functions and mechanisms are often deregulated in cancer. Aberrant signaling at cell-cell and cell-matrix adhesion sites often involves downstream mediators including Rho GTPases and tyrosine kinases. This review discusses these molecules as putative mediators of cellular crosstalk between cell-cell and cell-matrix adhesion sites, in addition to their attractiveness as therapeutic targets in cancer. Interestingly, inter-junctional crosstalk mechanisms are frequently typified by the way in which bacterial and viral pathogens opportunistically infect or intoxicate mammalian cells. This review therefore also discusses the concept of learning from pathogen-host interaction studies to better understand coordinated communication between cell-cell and cell-matrix adhesion sites, in addition to highlighting the potential therapeutic usefulness of exploiting pathogens or their products to tap into inter-junctional crosstalk. Taken together, we feel that increased knowledge around mechanisms of cell-cell and cell-matrix adhesion site crosstalk and consequently a greater understanding of their therapeutic targeting offers a unique opportunity to contribute to the emerging molecular revolution in cancer biology.

Targeted Macromolecules Delivery by Large Lipidic Nanovesicles Electrofusion with Mammalian Cells

Lipidic nanovesicles (so called liposomes) were one the earliest forms of nanovectors. One of their limits was our lack of knowledge on the delivery pathway of their content to the target cell cytoplasm. The present communication describes an efficient way to enhance the delivery. Pulsed electric fields (PEF) are known since the early 80’s to mediate a fusogenic state of plasma membranes when applied to a cell suspension or a tissue. Polykaryons are detected when PEF are applied on cells in contact during or after the pulses. Heterofusion can be obtained when a cell mixture is pulsed. When lipidic nanovesicles, either small unilamellar vesicles (SUVs) or large unilamellar vesicles (LUVs), are electrostatically brought in contact with electropermeabilized cells by a salt bridge, their content is delivered into the cytoplasm in electropermeabilized cells. The PEF parameters are selected to affect specifically the cells leaving the vesicles unaffected. It is the electropermeabilized state of the cell membrane that is the trigger of the merging between the plasma membrane and the lipid bilayer. The present investigation shows that the transfer of macromolecules can be obtained;i.e. 20 kD dextrans can be easily transferred while a direct transfer does not take place under the same electrical parameters. Cell viability was not affected by the treatment. As delivery is present only on electropermeabilized cells, a targeting of the effect is obtained in the volume where the PEF parameters are over the critical value for electropermeabilization. A homogeneous cytoplasm labeling is observed under digitised videomicroscopy. The process is a content and “membrane” mixing, following neither a kiss and run or an endocytotic pathway.

Effects of Polysilane-Coating on Interface of Electrofusion Joints for Maintaining Strength

The fusion strength of electrofusion joints using the polyethylene (PE) pipe connection greatly depends on the amount of sand which adheres to the interface by wind and so on, because there is no flow of melted resin at the fusion interface on electrofusion joints. Therefore, it is necessary to develop a method to prevent the fusion strength from reducing even in the case of sand adhesion. In this study, the fusion interface coated with polysilane, a kind of silicon polymer, effectively prevented the reduction of the fusion strength even if contaminated by sand. It was found that it brought the improvement of the fusion strength since when there was polysilane on the fusion interface. PS deeply permeated the polyethylene layer and lowered the viscosity of polyethylene.

Sensing Traction Strain Induces Cell-Cell Distant Communications

Mechanobiology has been a highly recognized field in studying the importance of physical forces in physiologies at the molecular,cellular,tissue,organ and body-levels.Beside the intensive work focusing on the fine local biomechanical forces,the long-range force which can propagate through a relatively distant scale(in hundreds of micrometers and beyond)has been an intriguing topic with increasing attentions in recent years.The collective functions at cell population level often rely on cell-cell communications with or without direct contacts.Recent progresses including our own work indicate that the long-range biomechanical force propagating across scales far beyond single cell size may reserve the capability to trigger coordinative biological responses within cell population.Whether and how cells communicate mechanically in a distant manner remains largely to be explored.In respiratory system,the mechanical property of airway smooth muscle(ASM)is associated with asthma attack with prolonged contraction during airway hyper-responsiveness.In this work,we found that ASM cells rapidly self-assembled into a well-constructed network on 3D matrigel containing type I collagen(COL I),which required the collective functions and coordination of thousands of cells completed within 12-16 hours.Cells were assembled with aligned actin stress fibers and elongated nuclei.The assembling process relied on the long-range mechanical forces across the matrix to direct cell-cell distant interactions.We further found that single ASM cells could rapidly initiate multiple buds precisely pointing to neighboring cells in distance,which relied on cell traction force and force strain on the matrix.Beads tracking assay demonstrated the long-range transmission of cellular traction force to distant locations,and modeling of maximum strain distribution on matrix by finite element method predicted the consistency with cell directional protrusions and movements in experiments.Cells could sense each other in distance to move directionally on both non-fibrous matrigel and in much more efficient way when containing COL I.Cells recruited COL I from the hydrogel to build nearly identical COL I fibrous network to mechanically stabilize the cell network.Our results revealed that ASM cells can sense the traction strain transmitted through matrix to initiate distant communications and rapidly coordinate the network assembly at the population level through active cell-matrix interactions.As an interesting phenomenon,cells sound able to’make phone call’via the role of long-range mechanical force.In summary,this work demonstrated that long-range biomechanical force facilitates the collective functions of ASM cell population for network assembly.The cells reacted to traction strain on the matrix for distant communications,which resulted in directional budding and movement.Fibrous COL I had important roles in facilitating the efficiency of force transmission to induce the assembly and stabilizing the cell network.This work has helped advance the understanding of the feature andfunction of long-range biomechanical force at the cell population level.The observed high mechano-sensitivity of ASM cells might suggest a re-enforced feedback of enhanced contraction by excessive ASM under asthmatic condition.

Could gastrointestinal tumor-initiating cells originate from cell-cell fusion in vivo?

Tumor-initiating cells(TICs)or cancer stem cells are believed to be responsible for gastrointestinal tumor initiation,progression,metastasis,and drug resistance.It is hypothesized that gastrointestinal TICs(giTICs)might originate from cell-cell fusion.Here,we systemically evaluate the evidence that supports or opposes the hypothesis of giTIC generation from cell-cell fusion both in vitro and in vivo.We review giTICs that are capable of initiating tumors in vivo with 5000 or fewer in vivo fused cells.Under this restriction,there is currently little evidence demonstrating that giTICs originate from cell-cell fusion in vivo.However,there are many reports showing that tumor generation in vitro occurs with more than 5000 fused cells.In addition,the mechanisms of giTIC generation via cell-cell fusion are poorly understood,and thus,we propose its potential mechanisms of action.We suggest that future research should focus on giTIC origination from cell-cell fusion in vivo,isolation or enrichment of giTICs that have tumor-initiating capabilities with 5000 or less in vivo fused cells,and further clarification of the underlying mechanisms.Our review of the current advances in our understanding of giTIC origination from cell-cell fusion may have significant implications for the understanding of carcinogenesis and future cancer therapeutic strategies targeting giTICs.

基于Logistic增长和“cell-cell”传播的动力学模型

建立了一个考虑Logistic增长和“cell-cell”传播的动力学模型。运用微分方程的稳定性理论对模型进行全局动力学分析,并进行了数值模拟。

Electrofusion between Blastula Cells and Unfertilized Egg in Loach (Paramisgurnus dabryanus)

Electrofusion between blastula cells and unfertilized eggs in loach were investigated usingdielectrophoretic field where, under alternating sinusoidal electric field, blastula cells formed beads-like chain in close contact with the unfertilized egg and cell fusion occurred between eggs and thecells in tight contact with them. The nuclei ofblastula cells were brought into the cytoplasm of therecipient eggs, where they promoted the development of the fused eggs just like the zygote nuclei.But the development of the fused eggs was different from that of zygotes. Several nuclei might enterone and the same egg simultaneously and all of them could undergo division, resulting in severalblastomere after the first cleavage of the recipient egg. Before blastula stage, the embryo developingfrom the fused egg showcd irregular shape, but it was soon regulated and developed to a normalblastula which often continued its development into a normal individual. Cell/egg electrofusion cameto its highest fosion rate (80%) 8nd hatching rate (20%), with cell density at 1×10~3 cells/ml, Ca^(++)concentration at 10 mM, mannitol at 0.2 M and when the blastula cells were digested with 100μg/ml pronase E for 6-10 min at 20℃. The mechanism underlying development of electrofused eggsis discussed. As the result indicates, electrofusion might prove to be a promising biotechnology justas nuclear transplantation.

Characteristics of cell motility during cell collision

Quantitative examination of cellular motion and intercellullar interactions possesses substantial relevance for both biology and medicine.However,the effects of intercellular interactions during cellular locomotion remain under-explored in experimental research.As such,this study seeks to bridge this research gap,adopting Dictyostelium discoideum(Dicty)cells as a paradigm to investigate variations in cellular motion during reciprocal collisions.We aim to attain a comprehensive understanding of how cell interactions influence cell motion.By observing and processing the motion trajectories of colliding cells under diverse chemical environments,we calculated the diffusion coefficient(D)and the persistence time(τ),using mean square displacement.Our analysis of the relationship dynamics between D andτprior to the collisions reveals intricate and non-monotonic alterations in cell movements during collisions.By quantitatively scrutinizing theτtrend,we were able to categorize the cellular responses to interactions under different conditions.Importantly,we ascertained that the effect of cell interactions during collisions in Dicty cells emulates a classical sigmoid function.This discovery suggests that cellular responses might comply with a pattern akin to the Weber–Fechner law.

scRNA-seq transcriptomic profiling of irradiated mouse skin reveals altered cell types,pathways,and cell-cell interactions

Objective:To investigate the substantial changes in cell types,pathways,and cell-cell interactions occurring in the irradiation-induced alopecia and dermatitis(IRIAD)mouse model and to identify potential targets for patients experiencing skin adverse reactions to radiotherapy.Methods:Mice were irradiated at 15 Gy,targeting the head and neck region.After a 14-day interval,living cells were extracted from both wild-type(WT)mice and irradiated mice for single-cell RNA sequencing(scRNA-seq).The scRNA-seq data,retrieved from the GEO database(GSE201447),underwent stringent quality control using the Seurat(v4.3.0)R package.Cell type annotation relied on previously reported typical markers and CellMarker 2.0.Differentially expressed genes were calculated to perform gene ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)analyses.Cell-cell interactions were evaluated using the Cellchat R package.Results:The application of single-cell RNA sequencing(scRNA-seq)enabled a comprehensive characterization of the intricate cellular composition of both wild-type(WT)and irradiated mice skin.Remarkably,cells within irradiated mice skin exhibited a significant alteration in the intensity of cell-cell interactions compared to their wild-type counterparts.This change in interaction intensity was observed across various cell types,including fibroblast cells,endothelial cells,and dendritic cells.Importantly,these"interacting cells"shared common signaling pathways,notably the upregulation of the IL-17 pathway following irradiation.Conclusions:The modification of intercellular communication induced by irradiation primarily involves fibroblast cells,endothelial cells,and various types of immune cells.This investigation provides a novel perspective on potential targets and holds promise for enhancing the clinical management of IRIAD.

聚乙烯管焊缝超声检测工艺探讨

通过对聚乙烯管不同焊接工艺所形成的焊缝结构分析,了解相应的焊接缺陷产生的机理,针对各自形成的焊缝形状,选择不同的超声检测工艺。对电熔焊接接头检测,采用相控阵超声检测工艺,优化其灵敏度、扫查孔径和聚焦深度的工艺参数来提高检测效率;对热熔焊接接头采用TOFD检测方法,选择合适TOFD楔块材质和规格来提高检测分辨力。

A Single-Cell Landscape of Human Liver Transplantation Reveals a Pathogenic Immune Niche Associated with Early Allograft Dysfunction

Liver transplantation(LT)is the standard therapy for individuals afflicted with end-stage liver disease.Despite notable advancements in LT technology,the incidence of early allograft dysfunction(EAD)remains a critical concern,exacerbating the current organ shortage and detrimentally affecting the prognosis of recipients.Unfortunately,the perplexing hepatic heterogeneity has impeded characterization of the cellular traits and molecular events that contribute to EAD.Herein,we constructed a pioneering single-cell transcriptomic landscape of human transplanted livers derived from non-EAD and EAD patients,with 12 liver samples collected from 7 donors during the cold perfusion and portal reperfusion stages.Comparison of the 75231 cells of non-EAD and EAD patients revealed an EAD-associated immune niche comprising mucosal-associated invariant T cells,granzyme B^(+)(GZMB^(+))granzyme K^(+)(GZMK^(+))natural killer cells,and S100 calcium binding protein A12^(+)(S100A12^(+))neutrophils.Moreover,we verified this immune niche and its association with EAD occurrence in two independent cohorts.Our findings elucidate the cellular characteristics of transplanted livers and the EAD-associated pathogenic immune niche at the single-cell level,thus,offering valuable insights into EAD onset.

燃气聚乙烯管道电熔接头泄漏事故失效机理分析

针对聚乙烯管道电熔接头泄漏事故,对焊缝进行了相控阵超声检测,并通过缺陷处电熔接头拉伸剥离试验,进一步分析了焊接接头的失效机理。结果表明:管件的过焊,使得管件与套筒之间的电阻丝区发生错位,形成熔合区未完全熔合的状态,聚乙烯材料发生大量断链反应,造成本次泄漏事故。最后,对新建管道现场施工和检验给出了建议。

Role of Immune Cells and Non-immune Cells with Immune Functions in the Pathogenesis of ALI/ARDS

This review outlines the effects of different types of cells with immune function on acute lung injury(ALI)inflammation and the regulation of inflammatory responses between these cells via cell-cell interactions.It is expected to provide some possible strategies for the research and treatment of ALI and acute respiratory distress syndrome(ARDS).