Oral diseases present a global public health problem that imposes heavy financial burdens on individuals and health-care systems. Most oral health conditions can be treated in their early stage. Even if the early symptoms of oral diseases do not seem to cause significant discomfort, prompt treatment is essential for preventing their progression. Biomaterials with superior properties enable dental therapies with applications in restoration, therapeutic drug/protein delivery, and tissue regeneration. Graphene nanomaterials have many unique mechanical and physiochemical properties and can respond to the complex oral microenvironment, which includes oral microbiota colonization and high masticatory force. Research on graphene nanomaterials in dentistry, especially in caries, periodontitis therapy, and implant coatings, is progressing rapidly. Here, we review the development of graphene and its derivatives for dental disease therapy. Oral diseases constitute a public health problem worldwide with high prevalence (3.4 billion cases) and incidence (4.35 billion cases) . Oral diThe goal of dentistry is, through an achievable plan, to maintain or rehabilitate a person’s oral health, promoting general health. Filling, also called restoration, is the main treatment method for caries, which requires various materials, such as composite resins and adhesives. These materials are exposed to saliva, oral microbiota, high masticatory force, and abrasion, which can lead to failure of the treatment, the formation of secondary caries, fracture of the restoration, and microleakage caused by the shrinkage of the resin or the dissolution of adhesive ,8. MoreoApplications of nanotechnology have greatly contributed to the development of dentistry, and one of the more prominent aspects is the innovative development and application of nanomaterials in dental practice ,21. Grap2 -hybridized carbon atoms with the arrangement of a honeycomb lattice [Graphene, the thinnest and strongest material known, is a single atomic sheet made up of sp lattice ,26. It w lattice . Graphen lattice ,30,31,32 lattice ,34,35. N lattice ,28,36,37 lattice ,39,40,41Graphene oxide (GO) and reduced graphene oxide (rGO) are two main graphene derivatives. GO was first discovered in 1859 and presented many oxygen-containing functionalities, such as hydroxyl epoxy and carboxyl groups, which contribute to the covalent or noncovalent combination of GO with biomolecules and other nanomaterials ,43,44. GChemical vapor deposition, chemical-based methods, and micromechanical exfoliation of graphite are three main techniques used to synthesize graphene ,51. The The oxidization of natural graphene results in graphite oxide, followed by sonication exfoliation to produce GO, the reduction of which forms rGO . The cheRecently, graphene and its derivatives have given rise to great interest in the fields of biomedicine and dentistry. Similar to other inorganic nanomaterials, assessments of cytotoxicity are critical to their further clinical application. The biological toxicity of graphene and its derivatives has been systematically reviewed . The toxDecreasing the restoration failure caused by bulk or marginal fractures, as well as reducing the risk of secondary caries, to minimize the demand to replace restorations is a crucial goal of contemporary dentistry ,76. The Graphene-based materials have been introduced into dentistry to improve the performance of dental materials. Evidence has indicated that graphene can enhance the mechanical and physicochemical properties of biomaterials, being biocompatible and noncytotoxic in the form of few-layer graphene (FLG), generally 1 to 6 layers. Therefore, FLG should be an ideal material that can be incorporated into dental polymers, thereby increasing their strength and durability. Malik S et al.  fabricatIn addition to GICs, bioactive calcium silicate bone cement is widely used in dentistry for endodontic treatments such as the management of perforations, retrograde root filling, and pulp capping due to its strong sealing ability and mineralization inducing ability, but it still has some problems in clinical application, including a long setting time and unsatisfactory physico-mechanical properties. Dubey et al.  tried toUnlike GICs and bioactive calcium silicate bone cement, PMMA, known as “organic glass”, is widely used as a denture material due to its superior heat resistance, high strength, and low cost. However, it suffers from brittleness and poor impact resistance. A previous study demonstrated that incorporation of G–AgNp improved the flexural strength of PMMA resin . FurtherIn addition, GO could improve the performance of adhesive and silane primers and protect Ti substrates ,88,89. SStreptococcus mutans is a key cariogenic pathogen and can synthesize insoluble EPS using dietary sucrose, thereby facilitating bacterial adhesion–cohesion and accumulation on the tooth surface, subsequently promoting the formation of caries-causing acidogenic biofilms [S. mutans-targeting materials is of great importance for controlling the pathologic condition. Although widely explored biocides, such as chlorhexidine, present a strong ability to inhibit S. mutans, they also suppress the growth of beneficial bacteria, disrupting oral microbiota homeostasis [Dental caries is a disease that relies on cariogenic biofilms, which are highly organized microbial communities embedded in a cohesive matrix of extracellular polymers, mainly extracellular polysaccharide (EPS) . Streptobiofilms . Therefoeostasis . CurrentS. mutans in vitro [S. mutans, and the antibacterial properties improved with increasing FG content. When 4 wt% FG was added, the antibacterial rate of S. mutans reached 85.27%. Abundant evidence has demonstrated that graphene and its derivatives are promising anti-caries nanomaterials due to their impressive ability to inhibit cariogenic bacteria and prevent tooth demineralization [S. mutans using the 3--2,5-diphenyl tetrazolium bromide (MTT) reduced test, colony forming unit counting, growth curve observation, and live/dead fluorescent staining [S. mutans. Consistent with this result, the addition of 0.25% graphene nanoplatelets doped with silver nanoparticles into adhesives showed optimized antibiofilm properties against S. mutans without affecting the standard adhesion characteristics regarding bond strength, leakage expression and durability at the resin–dentin interface [S. mutans, graphene oxide–copper nanocomposites (GO–Cu) and graphene/zinc oxide nanocomposites (GZNC) could also reduce the biomass of the S. mutans biofilm and suppress cariogenic properties of S. mutans, such as acid production and glucan formation [Abundant evidence has demonstrated that graphene and its derivatives are promising anti-caries nanomaterials due to their impressive ability to inhibit cariogenic bacteria and prevent tooth demineralization ,97,98,99in vitro ,93,94,96lization . A studystaining  found thormation .S. mutans [S. mutans cells using GOPEI complexes, and a reduction in virulence-associated gene (gtfBCD and gbpB) expression due to AsvicR was observed. Both GO and AsvicR alone could significantly suppress biofilm formation and EPS production. In contrast, GOPEI–AsvicR exhibited much more remarkable inhibitory effects with respect to virulence-associated gene expression, biofilm aggregation, and EPS accumulation, which may be related to silencing the expression of the vicR gene and the physical effect of GO. They suggested preserving nanographene oxide with AsvicR RNA as a more effective strategy for dental caries management [S. mutans [Interestingly, there are studies that combine GO with new technologies to achieve its antibacterial effect . When io. mutans . Wu S et. mutans  developenagement  , pulp regeneration, and apical induction are the main and effective treatments for teeth with pulpitis or apical periodontitis. RCT is a fundamental step to remove infected tissue and pathogens in the root canal system. To reach the goals of RCT, filling materials used to occupy complex root canal systems are supposed to have the desired sealing ability, biocompatibility, and antibacterial properties. Persistent infection in confined areas of the root canal system is a main reason for root canal treatment failure and posttreatment apical periodontitis. Owing to the less accessible areas of the root canal system, mechanical debridement and chemical irrigation during RCT are often not enough to eliminate bacteria from the root canal system. Therefore, it is necessary to develop new strategies, such as agitation and activation methods, to improve efficacy.Enterococcus faecalis , Ag–GO particles and GO–ICG–PDT could effectively reduce the biovolumes of faecalis , which htic acid . These ftic acid . Recentltic acid . Akbari tic acid  fabricatPulp regeneration, a promising technique to treat pathological or iatrogenic dental pulp lesions caused by caries or pulpectomy, is expected to replace RCT in the future . In a piPeriodontitis is a chronic infectious disease of tooth supporting tissues, including gingiva, cementum, alveolar bone, and the periodontal ligament. Peri-implant diseases are inflammatory conditions that influence the soft and hard around dental implants. They are considered a special subtype of periodontal diseases, which could result in failures of dental implants. The primary goal of periodontitis and peri-implantitis treatment is to control infection and to prevent tissue destruction by removing multispecies biofilms on the root/implant surface and reducing tissue invasion.Porphyromonas gingivalis is regarded as a keystone periodontal pathogen, so it is the target bacteria for many new dental drugs and materials development research. GO nanosheets could effectively suppress the viability of P. gingivalis in a dose-dependent manner, stopping growth at a concentration of 40 μg/mL [Fusobacterium. nucleatum via a similar mechanism [P. gingivalis, F. nucleatum, and S. mutans [Aggregatibacter actinomycetemcomitans, P. gingivalis, and Prevotella intermedia). This is associated with ROS generation and the downregulation of biofilm genes  [S. mutans, F. nucleatum, and P. gingivalis reformation on implants well, and the effect was significantly better than that of single use [40 μg/mL . The mec40 μg/mL . GO nanoechanism . Thus, w. mutans . The und. mutans . GQDs cond inpA) . The comngle use . Taken tThe regeneration of lost tooth/implant-supporting tissues is an ambitious purpose of periodontal regenerative therapy. In stem cell-based therapy of dentistry regeneration, cells and/or growth factors often need to be delivered to the injured site by scaffolds. Periodontal ligament stem cells (PDLSCs) are readily available mesenchymal stem cells (MSCs) with promising applications in regenerative therapy. GO-based materials, such as GO combined with fibroin, GO-coated Ti substrates and GO-applied scaffolds, are regarded as promising biomaterials for tissue engineering due to their biological compatibility and bioactivity to promote the proliferation of PDLSCs ,122,123.Several lines of evidence have suggested the involvement of rGO in the proliferation of stem cells ,125. ForDental implants are now a common way to repair permanent tooth loss caused by oral diseases, especially dental hard tissue diseases and periodontitis. Ti is the most commonly used material in implantology, mainly due to its good biological compatibility. However, it has been demonstrated to release alloy particles and ions into peri-implant tissues, which could result in bone loss and osseointegration failure of the implant. Osseointegration is the “gold standard” used to assess the success of implants, and the interactions between the material surface and cells can be regulated by the characteristics of an implant surface. Therefore, considerable efforts have been made to optimize the properties of Ti implant surfaces to improve osteointegration. GO is regarded as a promising candidate to functionalize the surfaces of implants for the regulation of the interactions between implant surfaces and cells, and several groups have explored whether and how GO coatings of dental implants could improve osseointegration .In general, GO coatings can significantly enhance bone marrow mesenchymal stem cell (BMSC) adhesion, spreading, proliferation, and osteogenic differentiation in vitro and promote bone-implant osseointegration in vivo . More neInterestingly, GO was also studied as an efficient carrier to deliver therapeutic proteins, such as BMP-2 and SP . La W etS. mutans supplemented with sucrose for 8 days [Candida albicans biofilm growth and removal [Ti implants modified with graphene nanocoating (GN) can maintain their quality and electrochemical and structural integrity under biologically relevant stresses such as microbial-rich environments and inflammatory macrophages. Specifically, Ti implant surfaces with GN exhibited higher polarization resistance and lower corrosion rates after exposure to r 8 days . GN cove removal . However removal . In addi removal . This ma removal . Further removal .MPCR-TNZ, the multipass caliber-rolled Ti alloy of Ti13Nb13Zr, has been reported to have strong fatigue characteristics and high mechanical strength. For further applications in dental practice, Jung HS et al. obtained DEX/rGO–MPCR-TNZ by modifying the MPCR-TNZ surface with rGO followed by the loading of osteogenic DEX via π-π stacking on the graphitic domain of rGO. They found that DEX/rGO–MPCR-TNZ significantly stimulated MC3T3-E1 cell growth and osteoblast differentiation, showing remarkable expression of osteogenic markers, including Runx2, OPN, OCN and Col-1. rGO–MPCR-TNZ also exhibited an increase in calcium nodule deposition and ALP activity. The prototype implant of rGO–MPCR-TNZ was successfully implanted onto an artificial bone block with mechanical performance and structural characteristics similar to those of a jawbone, and the rGO on the surface remained stable even after implantation, confirming the feasibility of rGO–MPCR-TNZ for applications in clinical dentistry. Shin YC et al.  preparedTaken together, to date, there have been studies using graphene, GO, and rGO as coatings for Ti implants. Evidence from both in vitro and in vivo studies has suggested that GO coatings are beneficial for peri-implant bone formation and stabilization. The rGO coating also showed the potential to promote osteoblast differentiation; however, the pro-osteogenic effect of the rGO coating needs further investigation in vivo. Although graphene coatings have good stability in bacteria-rich and inflammatory environments, the ability of graphene coatings to cope with mechanical wear needs to be improved, and their osseointegration is unclear. Here, we summarize the applications of graphene and its derivatives mentioned above in dentistry for reference by subsequent researchers .S. mutans and P. gingivalis, which is beneficial for caries and periodontal disease therapy, as well as the success of implantation. However, many materials have this effect, and the current opinion is that dental caries and periodontal disease are not infections caused by a single pathogenic bacterium but result from oral microbiota dysbiosis and the imbalance of oral microbiota and host interactions. If graphene and its derivatives can selectively inhibit harmful bacteria without influencing beneficial bacteria, they will become a modern and powerful weapon for dentistry. Moreover, although graphene and GO can promote the differentiation and proliferation of DPSCs and PDLSCs, which is beneficial to the regeneration of dental pulp and periodontal tissues, whether they can act on specific genes or proteins to precisely regulate the host immune inflammatory response and maintain the balance between oral microbes and the host remains to be studied. In addition, the development of dentistry is inextricably linked to the delivery of biological drugs, including peptides, monoclonal antibodies, and nucleic acids. Graphene and its derivatives have shown great promise in the development of drug delivery systems, especially the delivery of drugs for targeted cancer therapy. However, there are currently only several lines of evidence that GO can be applied to oral biopharmaceutical delivery.Taken together, graphene and its derivatives can improve the mechanical properties of dental materials and present good biocompatibility; it may be an ideal material for caries filling, especially for severe caries filling. Existing evidence indicates that graphene and GO can inhibit the growth of both


TARDBP gene, is an RNA-binding protein with fundamental RNA processing activities, and its loss-of-function (LOF) has a central role in the pathogenesis of amyotrophic lateral sclerosis (ALS) and frontotemporal lobar degeneration (FTLD). TARDBP mutations are postulated to inactivate TDP-43 functions, leading to impaired RNA processing. However, it has not been fully examined how mutant TDP-43 affects global RNA regulation, especially in human cell models. Here, we examined global RNA processing in forebrain cortical neurons derived from human induced pluripotent stem cells (iPSCs) with a pathogenic TARDBP mutation encoding the TDP-43K263E protein. In neurons expressing mutant TDP-43, we detected disrupted RNA regulation, including global changes in gene expression, missplicing, and aberrant polyadenylation, all of which were highly similar to those induced by TDP-43 knock-down. This mutation-induced TDP-43 LOF was not because of the cytoplasmic mislocalization of TDP-43. Intriguingly, in nonneuronal cells, including iPSCs and neural progenitor cells (NPCs), we did not observe impairments in RNA processing, thus indicating that the K263E mutation results in neuron-specific LOF of TDP-43. This study characterizes global RNA processing impairments induced by mutant TDP-43 and reveals the unprecedented cell type specificity of TDP-43 LOF in ALS/FTLD pathogenesis.Transactivating response element DNA-binding protein of 43 kDa (TDP-43), which is encoded by the  TARDBP cause ALS and FTLD; thus, these mutations are considered to induce TDP-43 LOF. Here, using a human induced pluripotent stem cell (iPSC)-based model, we found that the pathogenic K263E mutation disrupted RNA processing events in neurons, reflecting TDP-43 LOF. Interestingly, in contrast to neuronal cultures, mutated TDP-43 remained functional in nonneuronal cells, including iPSCs and neural progenitors. This finding indicates an unprecedented neuron-specific LOF of TDP-43 that potentially accounts for the CNS-selective lesions in patients with ALS/FTLD.Altered RNA metabolism induced by transactivating response element DNA-binding protein of 43 kDa (TDP-43) loss-of-function (LOF) has been suggested to be a core disease mechanism in amyotrophic lateral sclerosis (ALS) and frontotemporal lobar degeneration (FTLD). Pathogenic mutations in TARDBP gene encoding TDP-43 in patients with ALS and FTLD  is a neurodegenerative disorder caused by the degeneration of upper and lower motor neurons, leading to a progressive loss of motor function. Frontotemporal lobar degeneration (FTLD) is the second most common type of dementia after Alzheimer’s disease and is characterized by progressive atrophy in the frontal and temporal lobes and by personality and behavioral changes. Transactivating response element DNA-binding protein of 43 kDa (TDP-43) was identified as a major component of pathologic inclusions found in the vast majority of patients with ALS and ∼50% of patients with FTLD . Moreoveand FTLD . These fand FTLD .STMN2, which is also observed in patients with ALS/FTLD  expressing TDP-43K263E did not mimic TDP-43 LOF. This study characterizes global RNA processing impairments induced by mutant TDP-43 and indicates that mutant TDP-43 exhibits LOF in a cell type-specific manner.In this study, using human iPSC-based models, we examined the K263E variant of TDP-43, which was initially identified in a patient with FTLD , with a 4 cells/well in an iMatrix 511 -treated six-well plate; 10 μm Y27632  was only added for the first day. Culture media were changed every other day.Human iPSCs 201B7;  were maiTARDBP were as follows: 5′-ATTGTGCTTAGGTTCGGCAT-3′and 5′-AATAGACAGTTAGAAAGAAG-3′. Both sequences were synthesized and ligated after the U6 promoter. The double-stranded targeting vector (TV) harboring the K263E mutation (c.A > G) together with the silent mutations at S258 (c.C > G) and S273 (c.A > T and c.G > C) flanked by 1-kbp homology arms was assembled using the NEBuilder HiFi DNA Assembly kit (New England BioLabs). iPSCs were electroporated with the Cas9 (D10A) expression vector, both sgRNA vectors, and TV at a ratio of 1:1:2 and verified as containing the biallelic K263E mutant using Sanger sequencing.CRISPR-Cas9-mediated homologous recombination was performed by GenAhead Bio. The sgRNA sequences targeting 5 cells/well in StemFit AK02 N medium. After 3 d, neural induction was initiated by changing the medium to neural induction medium  with 150 nm LDN193189 (StemRD), 5 μm SB431542 (Tocris), and 3 μm IWR1e . On day 6, the cells were dissociated into single cells using Accutase  and seeded onto poly-L-ornithine-coated and laminin-coated 12-well plates at a 1:1–1:2 ratio. On day 12, the cells were dissociated again, and 8 × 105 cells seeded in each well of poly-L-ornithine-coated and laminin-coated six-well plates and cultured in neuronal medium  with 20 μm DAPT (Sigma). On day 18, DAPT was removed, and 10 ng/ml BDNF (Alomone Labs), 10 ng/ml GDNF (Alomone Labs), and 1 μm PD0332991 (Sigma) were added. The medium was changed every 3 d.Neuronal induction of iPSCs was performed using a previously described method , with slTotal RNA was isolated from iPSCs and NPCs on day 12 and from neurons on day 36 with the RNeasy Mini kit (QIAGEN) with DNase I treatment. The quality of RNA  was assessed by Agilent 2100 Bioanalyzer (Agilent). The indexed cDNA libraries were prepared using the TruSeq stranded mRNA Library Preparation kit (Illumina) and sequenced using a NovaSeq6000 (Illumina) to obtain 150-bp paired-end reads at Macrogen. RNA-seq datasets of TDP-43 knock-down experiments from previous reports p values and a cutoff of 0.25 for the log2 fold change ratio. Principal component analysis (PCA) was performed using vst transformation of estimated counts based on intersections between DEG lists of two independent TDP-43 knock-down experiments from the studies by Salmon  was usedp values. Differentially spliced intron clusters were analyzed from the HISAT2-aligned data without existing isoform annotations by LeafCutter . Salmon index was prepared using this 3′ UTR sequence. 3′ UTR isoform usage was then quantified using “salmon quant” and “qapa quant.” PASs with TPMs >5 were retained for further analysis. For each gene, gene-level relative PAS usage was summarized using a metric Ψ  was evaluated, and gene-level PAS usage was summarized using the following formula:QAPA  was usedmetric Ψ . Each PAThe 3′ end-seq datasets from a previous report  were dowACTB levels. Data were analyzed using the comparative (ΔΔCt) method. The primers used for qPCR were as follows: ACTB, forward 5′-TGAAGTGTGACGTGGACATC-3′, reverse 5′-GGAGGAGCAATGATCTTGAT-3′; FOXG1, forward 5′-CCCGTCAATGACTTCGCAGA-3′, reverse 5′-GTCCCGTCGTAAAACTTGGC-3′; OCT4, forward 5′-GACAGGGGGAGGGGAGGAGCTAGG-3′, reverse 5′-CTTCCCTCCAACCAGTTGCCCCAAAC-3′; PAX6, forward 5′-ACCACACCGGTTTCCTCCTTCACA-3′, reverse 5′-TTGCCATGGTGAAGCTGGGCAT-3′; TARDBP-ΔIntron7, forward 5′-TTCATCTCATTTCAAATGTTTATGGAAG-3′, reverse 5′- ATTAACTGCTATGAATTCTTTGCATTCAG-3′; TUBB3, forward 5′-ATTTCATCTTTGGTCAGAGTGGGGC-3′, reverse 5′-TGCAGGCAGTCGCAGTTTTCAC-3′; and UNC13A-CE, forward 5′-TGGATGGAGAGATGGAACCT-3′, reverse 5′-GGGCTGTCTCATCGTAGTAAAC-3′.cDNA was prepared by using a ReverTraAce qPCR RT kit (Toyobo). The qPCR analysis was performed with TB Green Premix Ex Taq (TAKARA) using a ViiA 7 real-time PCR system (Applied Biosystems) according to the manufacturer’s instructions. Values were normalized to Neurons cultured until day 36 were fixed with 4% paraformaldehyde for 15 min at room temperature and then washed three times with PBS. After an incubation with blocking buffer  for 30 min at room temperature, the cells were incubated overnight at 4°C with primary antibodies at the following dilutions: TDP-43  and TUBB3 . The cells were again washed three times with PBS and incubated with secondary antibodies conjugated to Alexa Fluor 488 or 555 (Life Technologies) and Hoechst 33342 (Dojindo Laboratories) for 1 h at room temperature. After three washes with PBS and one wash with distilled water, the samples were mounted on slides and examined using an LSM-710 confocal laser scanning microscope (Carl Zeiss). Line-scan analysis was performed using ImageJ software. The resulting values were normalized to the maximum intensity. For the analysis of TDP-43 localization, TUBB3 staining was used to determine the cell body as the region of interest, and then Pearson’s correlation coefficient was calculated for TDP-43 and Hoechst staining using the Coloc2 plugin.GSE195689 (201B7 iPSC) and GSE196144 (otherwise).All the sequencing data have been deposited in the NCBI’s Gene Expression Omnibus and are accessible through GEO Series accession number K263E protein  and pseudo-alignment by Salmon (E). F, The variance of DEGs in neurons derived from TDP-43K263E iPSCs relative to those from wild-type iPSCs. Download Figure 1-1, EPS file.Quality of RNA-seq data. K263E iPSC-derived neurons . I, Percentages of cryptic splicing of UNC13A based on junction spanning read counts in #One sample in Melamed et al. contained no reads spanning exon 20–exon 21 junction, and was excluded for the splicing rate quantification. Download Figure 2-1, EPS file.STMN2 but also in various gene targets, including PFKP, DNAJC5, KALRN, SYT7, and UNC13B, all of which have been shown to be misspliced in TDP-43 knock-down models  that increase the efficiency of UNC13A cryptic splicing on TDP-43 loss (Extended Data UNC13A in these cells is expected to be drastically enhanced by TDP-43 LOF, which supports the second possibility. Indeed, RT-qPCR analysis revealed that UNC13A cryptic splicing was reproducibly increased in TDP-43K263E neurons (Extended Data UNC13A missplicing was actually occurring, but RNA-seq analysis was not sensitive enough to detect it. The low sensitivity of RNA-seq analysis to detect UNC13A missplicing was also supported by the finding that analyses using the data from UNC13A missplicing in only a subset of samples (Extended Data The exon usage analysis is based on existing isoform annotations, but disease-relevant missplicing often occurs in unannotated introns. We performed an annotation-free differential splicing analysis to characterize global changes in intron splicing . We deten models . On the P-43 LOF . To furtK263E and TDP-43 knock-down, TARDBP expression was upregulated in TDP-43K263E neurons (TARDBP expression was autoregulated by TDP-43 itself; namely, the binding of TDP-43 to its own mRNA changes the processing of the 3′ UTR, finally leading to a decrease in the mRNA level (TARDBP mRNA in TDP-43K263E neurons (K263E neurons compared with wild-type neurons (TARDBP mRNA, leading to the collapse of autoregulation.While gene expression profiling and splicing analyses indicated the similarity between TDP-43 neurons D. TARDBPNA level . Indeed, neurons B, sugges neurons , and gen neurons . Genes w neurons C. The us neurons D. These K263E neurons (PPP2R2D was preferentially used in wild-type neurons; on the other hand, TDP-43K263E neurons used the lengthened 3′ UTR (SMC1A was shifted similarly, but in the opposite direction (K263E affects RNA 3′ end processing of various genes because of the impaired TDP-43 function.TDP-43 regulates not only its own mRNA but also a broad range of 3′ end processing events . Thus, w neurons E. The prd 3′ UTR F. The 3′irection F. These irection . CollectK263E neurons. TDP-43 functions mainly in the nucleus, and its redistribution from the nucleus to the cytoplasm has been recognized as a pathologic hallmark of ALS and FTLD, suggesting that the pathogenic mechanism is associated with the loss of nuclear TDP-43 function (K263E neurons, we observed signals for TDP-43 immunostaining primarily in the nucleus (K263E neurons (Based on our results, the LOF of TDP-43 has been suggested in TDP-43function . Thus, w nucleus A,B. Pear nucleus C. In add nucleus , TDP-43  neurons A,B. TherK263E were also present in cell types other than neurons (Extended Data TARDBP was expressed at similarly high levels among iPSCs, NPCs, and neurons (TARDBP expression in TDP-43K263E iPSCs and NPCs compared with wild-type cells (K263E iPSCs and TDP-43 knock-down ESCs (K263E iPSCs and TDP-43 knock-down ESCs was also evident in the differential exon usage analysis, in which exon usage fold changes induced by TDP-43K263E were quite distinct from those induced by TDP-43 knock-down (POLDIP3 exon 3 was only observed in TDP-43 knock-down ESCs but not in TDP-43K263E iPSCs (TARDBP only in neurons; in contrast, this splicing event was rarely affected in TDP-43K263E iPSCs and NPCs (PPP2R2D mRNA, RNA-seq and 3′ end-seq revealed that the 3′ UTR was lengthened in TDP-43 knock-down ESCs, but TDP-43K263E iPSCs did not exhibit this PAS switch (K263E did not mimic the RNA processing impairment induced by TDP-43 depletion. This result is the opposite of the close relationship between the K263E mutation and LOF in neurons (We performed RNA-seq of iPSCs and NPCs expressing wild-type or K263E variant TDP-43 to test whether these RNA processing disruptions induced by TDP-43 neurons A. In conpe cells B. We reape cells , and theown ESCs C, suggesock-down D. Remark3E iPSCs E. Moreovand NPCs F. In theS switch G. Taken  neurons H.K263E (Human iPSC-based disease models have remarkable potential to clarify disease pathogenesis and to discover effective therapies because of the discrepancy between patients and animal models . PreviouK263E , substanK263E mutant indicate that K263E disrupts the RNA binding capacity of TDP-43 and enhances intranuclear TDP-43 aggregation or liquid shell formation (K263E, we did not observe aggregation or droplets of TDP-43 in our iPSC-derived neurons. This inconsistency is probably because of the difference in TDP-43 expression levels. Excess TDP-43 expression might enhance TDP-43 aggregation, but endogenous TDP-43 expression does not. This finding is supported by a neuropathological study of a patient with FTLD carrying the K263E mutation, in which intranuclear TDP-43 inclusions were not observed in the cerebral cortex (Previous reports have suggested several distinct but overlapping effects of mutations on TDP-43 functions, including cytoplasmic mislocalization, an increased tendency to aggregate, and a decreased RNA binding capacity . Cellulaormation . Althougl cortex .Additionally, cytoplasmic TDP-43 inclusions were indeed observed in this K263E carrier patient . In contK263E. TDP-43 LOF was only observed in neuronal cultures, whereas iPSCs and NPCs did not suffer from K263E mutation. The effect of TDP-43 mutation is often tested in nonneuronal cell lines, and comparisons among cell lines have been seldom studied. Thus, the findings for the TDP-43 mutant may need to be revisited in terms of the cell types tested. This cell type specificity is also important for the pathogenesis of ALS/FTLD. CNS-specific lesions in patients with ALS/FTLD do not match the observation that TDP-43 is expressed in various organs in the body (Our results strongly suggest the cell type-specific function of TDP-43the body . Neuron-


Consequently, the corresponding solid‐state lithium/LiFePO4 battery exhibits a high capacity of ≈146 mAh g−1 with a high capacity retention of 94.3% for up to 300 cycles.Elastomers offer attractive advantages over classical solid‐state electrolytes in terms of ensuring stable interfacial contact and maintaining fatigue durability, but the low ionic conductivity obstructs their practical applications in long‐life lithium metal batteries. In this work, rubber‐derived lithium‐conducting elastomer has been developed via sulfur vulcanization of nitrile butadiene rubber with a polymerizable ionic liquid to provide both high resilience and dramatically improved ionic conductivity. Owing to the chemically crosslinked network between rubber chains and ionic liquid fragments generated during vulcanization, the elastic lithium‐conductor achieves high resilience of 0.92 MJ m −3, superior fatigue durability of 1000 cycles at 50% strain, and high room‐temperature ionic conductivity of 2.7 × 10−4 S cm−1. This unique lithium‐conducting elastomer enables intimate contact and fast lithium‐ion transport, resulting in long‐life and high‐performance solid‐state lithium metal batteries.By virtue of a chemically crosslinked network via vulcanization approach, the rubber‐derived lithium‐conductor achieves high resilience of 0.92 MJ m Moreover, with the fast development of advanced energy storage systems with high‐energy density, mechanical properties of solid‐state electrolytes are becoming a key factor in withstanding repeated volume changes of electrodes  and large deformations of pouch cells. There are two main types of solid‐state electrolytes: inorganic solid electrolytes and solid polymer electrolytes (SPEs). The former include sulfides, oxides, nitrides, and phosphates, and can provide high ionic conductivities (10–4–10–2 S cm–1) close to or even higher than liquid electrolytes, but their brittle nature, high reactivity, and high interfacial impedance with electrodes hinder their practical applications. Compared to inorganic solid electrolytes, SPEs consisting of a polymer matrix and lithium salts possess significant processing advantage and superior interfacial contact, as well as comparable safety enhancements. Unfortunately, pristine polymer electrolytes with high mechanical strength usually have poor room‐temperature ionic conductivity of less than 10–5 S cm–1. The introduction of small molecule additives can plasticize or gelatinize polymer electrolytes and improve their ionic conductivity but significantly reduce the mechanical strength, causing irreversible plastic deformation or fracture at small strains and unstable interface contact with electrodes. Therefore, there is an urgent need for SPEs with both high mechanical performance and good ionic conductivity.The ever‐increasing demand for energy storage systems with high‐energy density, high safety, and long‐cycling life to power mobile electronics and electric vehicles has shifted the focus from organic liquid electrolyte‐based lithium batteries toward solid‐state batteries.1, 2 In the case of lithium metal batteries, elastomers can afford stable contact with electrodes even when metallic lithium anodes suffer from relatively infinite volume changes during deep charge/discharge cycling. Moreover, elastomers can be assembled with stretchable electrodes to form multifunctional batteries, thus powering wearable electronic devices such as healthcare devices, stretchable sensors, and implantable devices. To date, only a few lithium‐conducting elastomers with high ionic conductivities have been reported. For example, elastic poly‐butyl acrylate/succinonitrile (PBA/SN) and SiO2/poly(propylene oxide) (PPO) electrolytes were fabricated via polymerization‐induced phase separation and covalent‐dynamic hydrogen bonding dual‐crosslinking method, respectively, and the corresponding solid‐state lithium metal batteries operated well at room temperature. However, most reported elastomer‐based electrolytes are either poor at lithium conducting or suffer mechanical deterioration due to gelation. Therefore, new approaches need to be developed toward a resolution of the dilemma between ionic conductivity and mechanical properties.Elastomers are an ideal choice for solid electrolytes in long‐life batteries due to their high resilience to accommodate repeated volume changes of advanced electrodes during operation.12 In –3) and long‐lasting cyclic durability (1000 cycles) at a high strain of 50%. Furthermore, the elastic NBR/IBIL hybrid electrolyte achieves a high room‐temperature ionic conductivity of 2.7 × 10–4 S cm–1 as a result of fast lithium‐ion pathways paved by IBIL fragments. Consequently, the NBR/IBIL hybrid electrolyte provides stable contact with the electrodes and fast lithium‐ion transport, resulting in the high performances of solid‐state lithium metal batteries, including long cycling life of 2000 h for the symmetrical battery and 300 cycles for the full battery.In this work, a rubber‐derived lithium conductor (denoted NBR/IBIL hybrid electrolyte) has been fabricated by vulcanization of nitrile butadiene rubber (denoted NBR) with a polymerizable ionic liquid imide denoted IBIL) in an attempt to resolve the above dilemma. The NBR/IBIL hybrid electrolyte inherits the elasticity and fatigue durability of NBR by virtue of a chemically crosslinked network of NBR chains and IBIL fragments generated via free radical addition reaction during vulcanization. The resulting lithium‐conducting elastomer possesses high resilience  imide, denoted LiTFSI). As schematically illustrated in e Figure , with a  , which is lower than the other NBR‐based electrolytes, suggesting that the introduction of IBIL fragments has resulted in a high degree of amorphousness and high local mobility of the polymer chain segments at room temperature. The width of the Tg peak is less than 15 K, which is consistent with the random vulcanization of NBR and IBIL and the absence of nano‐ or micro‐phase segregation. To investigate the lithium‐conducting mechanism of the NBR/IBIL hybrid electrolyte, temperature‐dependent conductivities were measured and compared with those of NBR and v‐NBR electrolytes , one order of magnitude higher than that of typical PEO‐based composite electrolytes (≈10–5 S cm–1), and comparable to that of previously reported elastic SiO2/PPO electrolyte. Additionally, NBR/IBIL hybrid electrolyte has a low Li‐ion transference number of 0.22  spectroscopy. The 7Li NMR peak of the hybrid electrolyte  has been introduced to quantify its elasticity, which is the maximum reversible stored energy in the material and can be given by the area under the linear stress–strain curve. Ur of the NBR/IBIL hybrid electrolyte (0.92 MJ m–3) is ≈2.2 times that of PBA/SN electrolyte (0.41 MJ m−3), ≈2.9 times that of SiO2/PPO electrolyte (0.32 MJ m−3), and much higher than the values for other reported electrolytes  cathode. The full battery's cyclic voltammograms (CV) were recorded at 0.5 mV s–1, and typical characteristic redox peaks of Fe2+/Fe3+ couple for LFP cathodes were detected at ≈3.7/3.2 V . In contrast, full batteries using NBR, v‐NBR, and NBR‐IBIL gel electrolytes failed rapidly  is comparable to the initial value, confirming the good cycling stability of the hybrid electrolyte. A Li/LFP battery with NBR/IBIL hybrid electrolyte and a high loading LFP cathode was also performed at 0.2 C (Figure –2, which is higher than that of a full battery with SiO2/PPO electrolyte at 0.2 C under 25 °C. The safety of solid‐state pouch cells using the NBR/IBIL hybrid electrolyte was further confirmed by abuse testing. The solid‐state pouch cell continues to power an LED at room temperature even when subjected to bending, puncturing, and cutting (Figure The electrochemical performance of the NBR/IBIL hybrid electrolyte in full batteries was further evaluated using metallic lithium foil as the anode and a standard LiFePO cathodes31 wery Figure , the fird Figure . When th3−3, long‐lasting fatigue durability, and high room‐temperature ionic conductivity of 2.7 × 10−4 S cm−1 to enable intimate contact and fast lithium‐ion transport, resulting in stable, long‐cycling, and high‐performance solid‐state lithium metal batteries. Given the rich variety, diverse structures, and modifiable properties of rubber materials, we believe this vulcanization method can be extended to explore a range of lithium‐conducting elastomers with highly tunable mechanical properties and good ionic conductivity for practical applications in solid‐state lithium metal batteries.In conclusion, a rubber‐derived lithium‐conducting elastomer has been fabricated using the vulcanization approach. By virtue of the chemically crosslinked network consisting of NBR chains and IBIL fragments generated during vulcanization, both mechanical performance and lithium‐ion transport of NBR/IBIL hybrid elastomer can be guaranteed. Hence, elastic NBR/IBIL hybrid electrolyte achieves high resilience of 0.92 MJ mThe authors declare no conflict of interest.Supporting InformationClick here for additional data file.


These printing technologies are now being applied in microbiology, with many recent examples where microbes and matrices are co‐printed to create bespoke living materials. Here, we propose a new paradigm for microbial printing. In addition to its importance for materials, we argue that printing can be used to understand and engineer microbiome communities, analogous to its use in human tissue engineering. Many microbes naturally live in diverse, spatially structured communities that are challenging to study and manipulate. 3D printing offers an exciting new solution to these challenges, as it can precisely arrange microbes in 3D space, allowing one to build custom microbial communities for a wide range of purposes in research, medicine, and industry. Many microbes naturally live in diverse, spatially structured communities, that are challenging to study and manipulate. 3D printing offers an exciting new solution to these challenges, as it can precisely arrange microbes in 3D space, allowing one to build custom microbial communities for a wide range of purposes in research, medicine, and industry. Proposed as early as 1945 in science fiction  embedded in human skin by outputting a fluorescence signal in response to the inducers , which is then converted by Saccharomyces cerevisiae into betaxanthin. By printing the two species in different spatial arrangements, the authors found that they could increase betaxanthin production by spatially segregating E. coli and S. cerevisiae  at the University of Oxford. K.R.F. is funded by a European Research Council Advanced Grant (787932) and a Wellcome Trust Investigator Award (209397/Z/17/Z).K.R.F. is a co‐founder of Postbiotics plus research LLC.


PBP1 reduces TDP-43 toxicity while overexpression enhances toxicity, led to the discovery that its human homolog, ATXN2, is associated with ALS risk. Thus, the yeast system has relevance to human disease. We now show that deletion of a new yeast gene, tip41Δ, likewise suppresses TDP-43 toxicity. We also found that TDP-43 overexpression and toxicity is associated with reduced autophagy. This is consistent with findings in other systems that increasing autophagy reduces TDP-43 toxicity and is in contrast to a report of enhanced autophagy when TDP-43 was overexpressed in yeast. Interestingly, we found that deletions of PBP1 and TIP41, which reduced TDP-43 toxicity, eliminated TDP-43′s inhibition of autophagy. This suggests that toxicity of TDP-43 expressed in yeast is in part due to its inhibition of autophagy and that deletions of PBP1 and TIP41 may reduce TDP-43 toxicity by preventing TDP-43 from inhibiting autophagy.When human TDP-43 is overexpressed in yeast it is toxic and forms cytoplasmic aggregates. The mechanism of this toxicity is unknown. Genetic screens for TDP-43 toxicity modifiers in the yeast system previously identified proteins, including PBP1, that enhance TDP-43 toxicity. The determination in yeast that deletion of  Yeast has been useful in the study of human proteins that form amyloid or prion-like aggregates associated with certain diseases ,4,5,6,7.PBP1, respectively reduces or enhances TDP-43 toxicity led to the discovery that PBP1′s human homolog, ATXN2, is associated with ALS risk. Research shows that an increase in the size of the polyglutamine expansions in ATXN2, within the normal range, is associated with enhanced risk for ALS  MATa his3Δ1 leu2Δ0 met15Δ0 ura3Δ0), and isogenic deletions from the yeast deletion library made in this strain, were obtained from Open Biosystems, Huntsville, AL; Cat # YSC1053 , and resolved on 10% SDS-PAGE followed by immunoblotting with α-GFP  to compare the level of uncleaved GFP-ATG8 and cleaved GFP, with α-TDP-43  to measure the level of TDP-43, and with α-Pgk  for an internal loading control. The ratio of cleaved GFP to uncleaved GFP-ATG8 measures autophagy. To compare the level of TDP-43 present in WT vs. deletion strains the internal PGK control was used. To determine the level of cleavage of GFP-ATG8 cells were grown in plasmid selective glucose media (SD -Leu -Ura -His), harvested, washed, normalized to 0.2 ODapamycin . Cells w600 into 2% galactose medium supplemented with 2% raffinose, 50 µM Cu++ and 1.6 nM FM4-64  and grown for 18 hrs with shaking at 30 °C to visualize cell membranes and vacuoles. Fluorescence was visualized with a Nikon Eclipse E600 fluorescent microscope (100× oil immersion) equipped with GFP and mCh filters. Cells were examined for GFP-ATG8 fluorescence, FM4-64 staining, and trypan blue staining. Cells with diffuse fluorescence in the cytoplasm without a concentration of fluorescence in the vacuole, or fluorescence in cytoplasmic foci even if accumulation of fluorescence in the vacuole, were scored as exhibiting inhibited autophagy. Cells with a concentration of fluorescence in the vacuole without punctate cytoplasmic foci, or live cells with no fluorescence, were scored as having undergone autophagy. Dead cells were detected with trypan blue staining seen in bright field.The cellular location of GFP-ATG8 determined by microscopy was also used to measure autophagy. Triple transformants verified in ICY1 and SSB1 did not affect toxicity associated with galactose induced overexpression of TDP43, while deletion of TIP41 had a big effect. Thus, we focused on TIP41. The effect of overexpression of TDP-43 on growth and cell viability was compared in yeast strain BY4741 and isogenic deletions of TIP41, PBP1 and ATG6 made in that strain background. These three strains were transformed with pGAL-TDP-43 (or its empty vector control) to allow for overexpression of TDP-43 on galactose, phER to allow the addition of 2 µM β-estradiol to induce overexpression of TDP-43 from pGAL-TDP-43 in dextrose media, and pCUP1-GFP-ATG8 to express GFP-ATG8 in the presence of 50 µM Cu++ to enable measurement of autophagy. Growth of spots of serial dilutions of at least four independent transformants was compared on plates and the fraction of dead cells was determined in liquid cultures  were used as controls. Autophagy was assayed by observing the breakdown or location of the autophagy reporter, GFP-ATG8, using both and immunoblotting  did not inhibit TDP-43 toxicity. Recalling that, in our conditions, deletion of PBP1 or TIP41 only effected autophagy in the presence of TDP-43 overexpression (Our data suggest that TDP-43 requires both PBP1 and TIP41 for it to inhibit autophagy and be toxic. The mechanism is unknown although their common effect on rescuing petites from inefficient mitochondrial import of proteins may not be the cause since other deletions with that phenotype (pression b, we conHere, we speculate on the last possibility, postulating TORC1 as the TDP-43 cellular target that causes toxicity and inhibits autophagy. In this scenario, in the absence of TDP-43, some TORC1 could be inactivated by being sequestered into stress granules or the TOROID structure  thereby


This study aimed to investigate whether non-radioactive assays can be used to quantitatively detect the lymphocyte proliferation <10% of normal as radioactive [3H]-thymidine, non-radioactive carboxyfluorescein diacetate succinimidyl ester (CFSE), and Ki-67 protein expressions were used to measure the lymphocyte proliferation as calculated using the stimulation index (SI), subtraction percentage, and proliferation index (FlowJo software). Normal references were established for comparison in the absence of parallel healthy controls.Radioactive [3H]-thymidine proliferation in those with T cell deficiency was compatible with CFSE- and Ki-67-stained lymphocyte percentages, and validated in patients with IL2RG, RAG1, and ZAP70 mutations. When calculating [3H]-thymidine <10% of normal lymphocyte proliferation, the threshold of parallel controls was more reliable than previously established normal references.Normal ranges of mitogen-stimulated lymphocyte proliferation were established as a SI of 15–267  with phytohemagglutinin (PHA) 5 μg/ml stimulation; 19–139  with concanavalin-A (ConA) 5 μg/ml stimulation; 7–53  with pokeweed mitogen (PWM) 0.1 ug/ml stimulation; 3–28  with candida 10 ug/ml stimulation; and 2–27  with bacille Calmette-Guerin (BCG) 0.02 ng/ml stimulation. The normalized CFSE-proliferation index was between 2.1 and 3.0. Although there was no significant correlation between these three assays in the healthy controls, the SI value for <10% [3H]-thymidine was more easily recognizable than that for non-radioactive CFSE and Ki-67. Even though the correlation was not significant, those identified to have <10% of normal proliferation by [3H]-thymidine could be consistently detected by CFSE and Ki-67, and consequently indicated for HSCT.The large quantitative value of radioactive [ Traditimulation . To prevmulation , 2 and n3H]-thymidine for the assessment of lymphocyte proliferation and reduce exposure to radiation.Theoretically, non-fluorescent CFSE passively diffuses into cells and is converted into highly fluorescent CFSE when the acetate groups are cleaved by intracellular esterases, subsequently releasing the succinimidyl ester group that reacts with intracellular amines to form fluorescent conjugates by a 2-fold decrease in cellular mean fluorescence intensity (MFI) , 2. Ki-63H]-thymidine was correlated with non-radioactive CFSE and Ki-67 assays with regards to lymphocyte proliferation. We then used these assays to investigate patients with profound T cell deficiency, speculated to what degree their lymphocyte proliferation was impaired, and assessed whether they met the indication for HSCT to reconstruct immunity.Patients with lymphocyte proliferation <10% of normal healthy controls are defined as having profound T cell deficiency, considered to be candidates for hematopoietic stem cell transplantation (HSCT) to reconstruct immunity and prevent them from life-threatening infections. In this study, we first established normal references and then investigated whether radioactive -thymidine as previously described (3H]-thymidine stimulation index (SI) was obtained from the formula: [stimulated count per minute (c.p.m.) – unstimulated or blank c.p.m.]/unstimulated c.p.m. × 100. A stock solution of CFSE  was prepared by dissolving CFSE in the DMSO at a concentration of 5 mmol/L. During the labeling process, 1 ml cell suspension was added to the bottom of a falcon tube, followed by the addition of 1:10 PBS-diluted CFSE stock solution. A final CFSE concentration of 5 μmol/L was loaded if the cell count was 2 × 106 cells/ml. After labeling, peripheral blood mononuclear cells (PBMCs) were placed in 96-well round-bottom plates with culture media under the indicated conditions, fixed with 2% paraformaldehyde , and then analyzed using a BD LSRII flow cytometer . In addition, the proliferation index of mitotic cells was calculated using the ModFit cell cycle analysis software (FlowJo).Lymphocyte proliferation was assessed escribed . The val6/ml PBMCs in the 96 round tube wells  cultured with blanks, mitogens, and antigens under the indicated conditions. The PBMCs in each well were then fixed, permeabilized (BD Biosciences), and stained with Ki-67 antibodies for 30 min (BD Biosciences). We determined the percentage of lymphocyte proliferation by subtracting those with stimulants and those with blanks only.We measured the kinetics of Ki-67 expression in lymphocytes using 10p < 0.05.Those with <10% of the normal range (an indication for HSCT) were defined by subtraction percentage and mean fluorescent intensity (MFI) from flow cytometry. Correlations between the different analysis methods were assessed using SPSS software version 17 . Statistical significance was set at 3H]-thymidine incorporation, SI, non-radioactive intracellular CFSE, Ki-67 percentage, and MFI were calculated  instead of using the original raw c.p.m. data to correlate the percentages and MFIs of CFSE- and Ki-67-stained lymphocytes and CFSE-proliferation index (FlowJo software) in the five age-matched healthy controls , ConA (5 ug/ml), and PWM (0.1 ug/ml), and the antigens candida (10 ug/ml) and BCG (0.02 ng/ml) to assess lymphocyte proliferation. Radioactive , RAG1 , and ZAP70  mutations, which are responsible for T cell deficiency, their PHA-stimulated lymphocyte proliferation as assessed by [3H]-thymidine incorporation was <10% compared with their parallel controls  supporting the indication for immune reconstruction by suitable HSCT, as our report with a good prognosis to date  combined T and B cell immunodeficiency (SCID or CID). Thus, we defined a threshold value <10% of normal proliferation from the healthy controls  as a potcontrols , which wcontrols . The acc to date .in vitro. If defective molecules involving activation pathways inhibit [3H]-thymidine nucleotide incorporation, incremental CFSE dilution, and nuclear Ki-67 entrance, the quantitative value of [3H]-thymidine measured using a beta-counter and flowcytometric percentages of CFSE and Ki-67 will increase less during stimulation, and this can explain recurrent opportunistic infections. In this study, we established standard normal ranges for comparison without the accompanying controls. The results showed that the quantitative value obtained with [3H]-thymidine incorporation was more sensitive and easily recognizable than that of CFSE and Ki-67 staining by flowcytometry, which depends more on skilled operators.Lymphocyte proliferation is a clinical screening method used to evaluate the lymphocyte response of a patient to mitogens and antigens Phytohemagglutinin-induced lymphocyte proliferation <10% in patients with severe T cell deficiency indicates HSCT for immune reconstruction. Otherwise, they will have increased susceptibility to opportunistic infections and die in infancy. Neonatal screening for T cell receptor excision circle (TREC) in the Guthrie card and consequently functional lymphocyte proliferation can identify patients with profound T cell immunodeficiency early  before s3H]-thymidine incorporation to detect lymphocyte proliferation is more sensitive and recognizable  without any detectable proliferation.Our results indicate that -thymidine); (2) by binding vital division-protein (Ki-67); and (3) by decreasing the concentration by 2-fold during each mitosis (CFSE). Therefore, the expressions of these three biomarkers could vary in the different stages of lymphocyte proliferation which may explain the non-significant correlation with each other. Second, the optimal conditions for the three biomarkers to quantify the strongest expression were different. Time-sequence analysis may reveal the adequate time-point to harvest. Third, those with <10% lymphocyte proliferation may have been misclassified as having >10% normal lymphocyte proliferation according to the previously established normal range in which the tested samples were obtained without parallel controls. Fourth, large-scale and time-sequence analysis studies are needed to investigate whether non-radioactive assays are correlated with radioactive lymphocyte proliferation, especially those without the profound impairment of lymphocyte proliferation.This study should be interpreted in light of its limitations. First, the three assays quantify lymphocyte proliferation using different mechanisms: (1) by incorporating a nucleotide , the National Science Council , and Taiwan Foundation for Rare Disorders (TFRD).The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher.


Late second language (L2) learners show translation priming from the first language (L1) to the second language (L1–L2), while L2–L1 effects are inconsistent. Late L2 learners also acquire the L2 after the L1 and are typically less dominant in the L2. As such, the relative contribution of language dominance and order of acquisition is confounded in these results. Here, Cantonese heritage and native speakers are tested in an auditory translation priming paradigm. As heritage speakers first learn Cantonese (L1) but later become dominant in English (L2), this profile allows for the potential dissociation of dominance and order of acquisition in translation priming. If order of acquisition is the primary factor, stronger priming is expected in the L1–L2 (Cantonese–English) direction; however, if dominance plays a stronger role, priming is expected in the L2–L1 (English–Cantonese) direction. Native speakers showed stronger L1–L2 priming, consistent with previous findings, while heritage speakers showed priming in both directions, and marginally larger L2–L1 priming. Treating language dominance as a continuous variable revealed that L1–L2 priming correlated with increased Cantonese dominance, while L2–L1 priming marginally correlated with increased English dominance. Collectively, these results suggest that both language dominance and order of acquisition help explain translation priming findings and bilingual lexical processing, generally. Overall, they invite a rethinking of the role of both variables in bilingual lexical access for speakers with different language dominance profiles. Hearing a word spoken in one language often activates similar words in the other language in bilingual speakers . ProficiTwo variables that characterise bilinguals are conflated in most late L2 learners: order of acquisition and relative language dominance . SpecifiTo examine the relative contributions of order of acquisition and language dominance, we tested a bilingual population, namely, heritage speakers, for whom their L2 is more dominant despite being acquired after the L1. Heritage speakers are often raised with an L1 that represents a  minority language at home but later became more dominant in the language of the wider community . AlthougIn nonselective models of bilingual lexical processing  were recruited both from the University of Toronto and Prolific (https://prolific.co/) and completed a web-based auditory translation priming lexical decision task. Participants reported no known linguistic, hearing, or neurological deficits and provided written informed consent prior to the experiment. Participants were compensated for their time either monetarily or with course credit. The experiment was approved by the Office of Research Ethics at the University of Toronto. Twenty-two participants who did not speak Cantonese were removed from further analysis. Thirty-nine participants were removed from the analysis for failing to achieve an accuracy rate greater than 75% on the lexical decision task. In total, 108 participants were included in the analysis .Cantonese–English bilinguals (SD = 1.1 years), followed by English . Participants were divided into two groups: Cantonese heritage and “native speakers” (see n = 72) and as native speakers if their BLP score was less than zero (n = 36). Supplementary Material.All participants were born to Cantonese-speaking parents and learned Cantonese first . All Cantonese words were disyllabic. English words varied between one and three syllables . The prime and target in each pair were always noncognates in distinct languages. In addition to these related translation pairs, each prime–target pair was matched with an unrelated, real-word Cantonese prime that was not a translation equivalent . Finally, these Cantonese–English pairs were matched with the corresponding English–Cantonese related and unrelated pairs . This produced translation priming pairs in the Cantonese–English direction  and in the English–Cantonese direction . Sixty real-word Cantonese–English related translation prime-target pairs were selected , and obtained from the English Lexicon Project  and 8 native speakers  who did not participate in the main experiment to complete a familiarity rating task. Both the Cantonese and English target words were included, and participants were randomly assigned to rate either the English words or the Cantonese words. Following the familiarity rating procedure in As our target population is heritage speakers, who are more dominant in English, it was important to ensure that all Cantonese items were familiar to our target sample; however, a suitable heritage speaker corpus that contained our experimental items was unavailable, making it difficult to control for lexical frequency. To this end, we recruited 16 heritage speakers  of The North Wind and the Sun. At the start of the block containing English targets, participants listened to the English version of The North Wind and the Sun, while at the start of the block containing Cantonese targets, they listened to the Cantonese version. The story was produced by the same speaker who recorded the experimental items. This was done to familiarise participants with the speaker’s voice and prepare them to make lexical decisions to the corresponding language of the target.At the start of the first block (irrespective of the target language of each block), participants were visually presented with instructions in English, and four practice trials with English primes and Cantonese targets (L2–L1 priming). Following the practice trials, participants were presented with either an English or Cantonese version  that appeared on the screen for 250 ms to indicate the start of the trial. Immediately after the fixation point, the auditory prime was presented to the participant. After a 500-ms inter-stimulus interval, the auditory target was presented to the participant, and participants made a lexical decision. Participants responded by pressing “1” on their keyboard if they heard a real word of Cantonese or English (depending on the block), or “0” if they heard a nonword of Cantonese or English. Both accuracy and reaction times (counted from the offset of the auditory target) were measured. The inter-trial interval was 1,000 ms. After the experiment was complete, participants completed the BLP questionnaire  and a shSupplementary Material. Overall, Cantonese heritage speakers were less accurate than native speakers ; however, this is largely due to the lower accuracy for heritage speakers to Cantonese nonwords . All other accuracy rates were above 90%. Only trials with correct responses were included in the reaction time analysis.All data aggregation and visualisation were conducted using the packages {dplyr}  and {ggpM = 556 ms, SD = 489 ms) responding faster than heritage speakers . There was also a main effect of Direction, with participants responding to the L2–L1 direction  faster than the L1–L2 direction . Finally, there was a main effect of Relatedness with participants responding slower to unrelated pairs  compared with related pairs . In addition to these main effects, there was a Direction × Relatedness interaction and a three-way Group × Direction × Relatedness interaction. For the Direction × Relatedness interaction, we observed a larger difference between the L1–L2 and L2–L1 directions in the unrelated trials  compared with the related trials . The descriptive statistics for the three-way interaction are presented in Reaction times were entered into a generalised linear mixed effects model with Gaussian errors  and a lo2(1) = 55.39, p < .001; L2–L1: χ2(1) = 72.55, p < .001, while native speakers showed priming only in the L1–L2 direction, χ2(1) = 43.04, p < .001, and not in the L2–L1 direction, χ2(1) = 1.65, p = .20. Priming magnitudes  by participant Group by Direction are presented in 2(1) = 11.77, p < .01, while for heritage speakers, marginally more L2–L1 priming (Δ = 166 ms) was observed compared with L1–L2 priming, Δ = 140 ms, χ2(1) = 2.86, p = .09. Finally, for the two-way interaction, the L2–L1 direction resulted in faster reaction times compared with the L1–L2 direction in the unrelated trials, χ2(1) = 18.45, p < .001, but not in the related trials, χ2(1) = 1.28, p = .26.To determine whether priming was observed in each priming direction for each participant group, as well as whether we observed more priming in the L1–L2 direction or the L2–L1 direction, a series of post hoc comparisons were conducted using the {phia} package . The Holz = –3.32, p < .001, τ = –0.23), and a marginal positive correlation between language dominance and priming in the L2–L1 direction . The more Cantonese dominant the participant was, the more L1–L2 priming was observed; the more English dominant the participant was, the more L2–L1 priming was observed.Order of acquisition and language dominance are still conflated in the native speakers, who showed stronger L1–L2 priming than L2–L1 priming. To dissociate these factors more clearly, as well as assess relative language dominance on a more granular level, participants’ BLP dominance scores were correlated with their priming magnitudes by direction see . KendallPrevious translation priming studies on late L2 learners have observed robust L1–L2 priming, whereas L2–L1 priming has been inconsistent and, traditionally, difficult to obtain in both visual L2–L1;  and audiWe found that heritage speakers showed marginally larger L2–L1 priming, while native speakers showed larger L1–L2 priming. Together, these results support the notion that language dominance drives translation priming, because both groups experienced stronger priming in the instances where their more dominant language acted as the prime . These findings are further supported on a more granular level in correlations between BLP dominance scores and priming magnitudes. Specifically, greater English dominance marginally correlated with greater L2–L1 priming, while greater Cantonese dominance correlated with greater L1–L2 priming. Although marginally more L2–L1 priming was observed compared with L1–L2 priming for heritage speakers, post hoc tests revealed that priming occurred in both directions. The same was not true for native speakers, however, for whom only L1–L2 priming was observed. The current findings are consistent with These results may be explained by recent models that posit nonselective access to an integrated lexicon. For example, in Multilink , lexicalMore broadly, Multilink formalises language use as language proficiency; however, language proficiency results from a variety of factors. Understanding the interaction between different bilingual factors as they contribute to language proficiency, overall, is necessary for a holistic account of bilingual lexical processing. Previous work has focused on late L2 learners and the role of order of acquisition in translation priming (see Previous translation priming studies have found robust L1–L2 priming and inconsistent L2–L1 priming; however, most of these studies test late L2 learners in the visual modality. Late L2 learner profiles conflate order of acquisition and language dominance: They both learn the L2 after the L1 and are less dominant in the L2. This makes disentangling the relative contribution of these two factors in translation priming difficult. To this end, the current experiment tested heritage Cantonese speakers in an auditory translation priming task. In heritage speakers, we observed marginally larger L2–L1 priming relative to L1–L2 priming, although heritage speakers showed priming in both directions. In native speakers, we only observed L1–L2 priming. Finally, English dominance positively correlated with L2–L1 priming and negatively correlated with L1–L2 priming. In sum, the current results suggest that both language dominance and order of acquisition are driving factors in the observed translation priming asymmetries, inviting a rethinking of lexical access in the context of bilingual speakers with various language dominance profiles.Click here for additional data file.Supplemental material, sj-pdf-1-qjp-10.1177_17470218221091753 for Language dominance and order of acquisition affect auditory translation priming in heritage speakers by Rachel Soo and Philip J Monahan in Quarterly Journal of Experimental PsychologyClick here for additional data file.Supplemental material, sj-pdf-2-csv-10.1177_17470218221091753 for Language dominance and order of acquisition affect auditory translation priming in heritage speakers by Rachel Soo and Philip J Monahan in Quarterly Journal of Experimental Psychology


The mixture of solid CO2 and Mg powder was filled in the pre-cooled sample tube and then impacted by a shock-driven cylindrical flyer tube. The impact generated a shockwave that propagated into the mixed precursor, inducing a chemical reaction between CO2 and Mg at a high shock pressure and high shock temperature. The recovered black powders were characterized via various techniques, confirming the presences of few-layer graphene. The mechanism is carefully shown to be that CO2 was reduced by Mg to form few-layer graphene under shock-induced high pressure and high temperature. By adding carbamide as an N source, this synthetic route was also applied to synthesize N-doped graphene nanosheets. Moreover, the yield and mass of the graphene materials in this study are up to 40% and 0.5 g, respectively. This study showed an efficient and easy-to-scale-up route to prepare few-layer graphene and N-doped few-layer graphene through shock synthesis.In this study, graphene and N-doped graphene nanosheets were synthesized through the shock-induced reduction of CO Graphene, the first two-dimensional 2D) material synthesized by Noveselov et al. , has beeD materia3 intercalated graphite powder with H2O2 to induce the decomposition of H2O2 to form O2 and the further expansion of graphitic layers to form graphene. In the arc-discharge method, Wu et al. [Multiple approaches have been documented for producing graphene materials, including the mechanical exfoliation approach, the oxidation reduction method, epitaxial crystal growth, chemical vapor deposition (CVD), the liquid exfoliation method, the arc-discharge method, the electrochemical exfoliation method, pulsed wire discharge, etc. Through the mechanical exfoliation of highly oriented pyrolytic graphite, mono-layer graphene with a lateral size of 1 mm was obtained successfully ,12. In tu et al.  obtainedu et al.  applied u et al. ,48.2 to useful carbon nanostructures. However, in a previous study, the yield of graphene recovered from shock-induced chemical reaction was 10%, approximately, due to the high sublimation rate of dry ice [In addition, the shock synthesis method can also be utilized to produce graphene . In the  dry ice .2, Mg fine powder, and carbamide (CH4N2O) with a high graphene yield. This work implies a potential large-scale production of graphene materials through carbon fixation using the shock synthesis method.In this work, we optimized the shock synthesis method to prepare few-layer graphene (FLG) and N-doped FLG by using CO2. Subsequently, the sample tube and cylindrical flyer tube were fixed on the cylindrical steel base with accessorial fixation of ring-like support to keep a gap of 2 mm between the sample tube and flyer tube. Then, the apparatus was assembled according to 2 and HCl, and dried using a vacuum-frozen drier for further characterization.To perform the shock treatment on precursors and for the recovery of the graphite samples, a cylindrical shock recovery apparatus was applied, as shown in 2 as an oxidant and carbon source, Mg as a reductant, and carbamide as a nitrogen source. The solid CO2 and Mg powder were mixed at a molar ratio of 1.1:2 in a mixing chamber cooled by liquid nitrogen. For the experiment to synthesize N-doped graphene, the carbamide is added in the precursor as a nitrogen source with the molar ratio 1:0.1 (solid CO2:carbamide). The carbamide powder was firstly mixed with Mg powder and then subsequently mixed with solid CO2 in the mixing chamber cooled by liquid nitrogen. The mass of the sample tube before and after filling it with the precursor were measured to obtain the mass of precursor powder (m0). The detailed experimental conditions and calculated shock pressure were listed in In these shock synthesis experiments, the mixed precursor consists of solid CO2 F20 S-Twin transmission electron microscope at an accelerating voltage of 200 kV to analyze the micromorphology of the recovered samples. Field emission scanning electron microscopy (SEM) observations were performed using a Hitachi S-4800 at an accelerating voltage of 5–15 kV. A LabRAM Aramis Raman spectrometer equipped with a He–Ne laser (excitation wavelength of 633 nm) was carried out to record the Raman spectra of recovered samples. The chemical composition and bonding states of recovered samples were probed by X-ray photoelectron spectroscopy (XPS) using a Thermo ESCALAB 250 Xi spectrometer with monochromatic AlK (1486.6 eV) X-ray sources. The recorded XPS spectra of all recovered samples were fit using a Gaussian–Lorentzian peak with non-linear Shirley background correlation. In addition, the samples for the characterization of SEM and Raman were separated in distilled water via ultrasonic dispersion and then dried on a Si plate.An X-ray Diffractometer (XRD)  was applied to record the XRD patterns of recovered samples with Cu Kα radiation (k = 0.15406 nm) and a step size of 0.0330° (2θ) from 10–90°. Transmission electron microscopy (TEM) and high-resolution TEM (HRTEM) observations of recovered samples were performed with a FEI Tecnai GThe representative SEM images  of the F2, etc., also indicates the high purity of the recovered FLG samples after the purification process.2 carbon are observed, including the D band (1332 cm−1), G band (1581 cm−1), and 2D band (in a range of 2663~2687 cm−1) in each spectrum. Through the spectra, the intensity ratios of the 2D band to the G band (I2D/IG) and the D band to the G band (ID/IG) are calculated to estimate the graphitic layers of the recovered samples [2D/IG values of the recovered samples are 0.71–1.13  and high pressure (14~18 GPa). These extreme conditions sublimate solid CO2 rapidly to form a high-density CO2 fluid in an enclosed sample tube with a small amount of CO and O2 formed by the partial decomposition of CO2. The high density of CO2 fluid also contributes to the reaction rate of the mixed precursor. Furthermore, shockwave propagation through the precursor powder is much higher than the combustion rate of Mg and CO2, leading to the ultra-high reaction rate of the mixed precursor in microseconds. Under this condition, the reduced free C atoms and oxidized MgO separate uniformly in the sample tube. Consequently, the free C atoms deposit on the surface of the adjacent MgO powder to form few-layer graphitic layers during the subsequent ultra-fast cooling process. After the purification process, the few-layer graphitic layers remain as FLG.The above characterizations confirm that shockwave action is an effective method to produce graphene materials. The formation mechanism of FLG from shock-treated precursor powder consisting of a mixture of solid COMoreover, the ultra-fast reaction rate, led by the shock loading and the uniform separation of free C atoms and MgO powder, also contributes to the uniform deposition to form FLG with the same orders of thickness and lateral size. Note that the higher pressure and higher temperature also contribute to the crystallization process of graphene to form graphene with a higher crystallinity considering the transient duration of this crystallization process. With respect to sample No. 3, the carbamide molecules decompose to produce free C atoms and N atoms under extreme high pressure and temperature during shock loading. These C atoms and N atoms also deposit on MgO surface, forming graphene and inducing the nitrogen doping during the deposition process to form N-doped graphene.2’s sublimation rate during experimental operation before detonation, leading to a high-efficiency preparation of graphene materials with a higher yield compared with previous studies. The graphene yield of samples Nos. 1–3 are in the range of approximately 70~80%. The formed N-doped graphene powder may have potential applications in the preparation of outstanding metal-free electrodes, which has been investigated using the specimen synthesized by the detonation method in our previous research.In addition, the pre-cooling treatment of the sample tube using liquid nitrogen efficiently inhibits the CO2 and Mg powder (with carbamide as N source). The shockwave technique provides an efficient carbon fixation route to transform CO2 into useful graphene materials. Experimental results confirm that shockwave action can induce the sublimation of solid CO2 to form dense CO2 fluid and further trigger the redox reaction between CO2 and Mg to produce FLG nanosheets within microseconds during the shock loading process. By adding carbamide as a N-doping agent, N-doped graphene can be obtained with a similar approach in one-step reaction route. The experimental results also imply that the increase in shock pressure and temperature is conducive to the crystallinity of the formed FLG and N-doped FLG though this shock-induced carbon fixation method.In this study, graphene and N-doped graphene materials are obtained through the shock-induced chemical reaction of CO


The first principles calculation and electrochemistry characteristic of the functional graphene films were performed on Materials Studio software and an electrochemical workstation, respectively. It is found that the nanostructures of functional graphene films with g-C3N4 and F-doped groups were significantly transformed. The introduction of fluorine atoms led to severe deformation of the g-C3N4 nanostructure, which created gaps in the electrostatic potential of the graphene surface and provided channels for electron transport. The surface of the roving fabric substrate covered by pure graphene is hydrophilic with a static contact angle of 79.4°, but the surface is transformed to a hydrophobic state for the g-C3N4/graphene film with an increased static contact angle of 131.3° which is further improved to 156.2° for CF2-modified g-C3N4/graphene film exhibiting the stable superhydrophobic property. The resistance of the electron movement of CF2-modified g-C3N4/graphene film was reduced by 2% and 76.7%, respectively, compared with graphene and g-C3N4/graphene.The physicochemical properties of functional graphene are regulated by compositing with other nano-carbon materials or modifying functional groups on the surface through plasma processes. The functional graphene films with g-C Graphene with a two-dimensional nanostructure possesses unique optical, electrical, and mechanical properties, which helps it work as a popular functional material ,2,3. The3N4) with high conductivity and photocatalytic activity to construct graphene and carbon nitride nanocomposite photocatalytic materials, which can thus not only retain their excellent properties but also produce synergistic effects to improve electrochemical activity [3N4 form a highly delocalized π conjugated system with sp2-C hybridization, which exhibits a suitable semiconductor band edge position and meets the thermodynamic requirements of photo-hydrolysis of aquatic hydrogen to produce oxygen. Dong [3N4/graphene/g-C3N4 sandwich heterostructure using density functional theory with van der Waals correction. The results showed that the band gap of the sandwich heterostructure could be as high as 106 meV without strain while maintaining high carrier mobility. Wu [3N4/graphene nanocomposites using first principles calculations. The calculation results showed that the buckle graphene/g-C3N4 nanocomposite was more stable than the planar one, and graphene as a substrate of graphene/g-C3N4 nanocomposite can obviously loosen the buckle degree of g-C3N4 to stabilize electron configuration. The main synthesis methods of g-C3N4 are mainly divided into solid phase reaction method, solvothermal method, electrochemical deposition method, and thermal polymerization [3N4 one-step preparation by CVD is rarely reported. In addition, the activity of nano-carbon-based materials can also be adjusted by doping atoms , so as to achieve the purpose of adjusting the charge distribution, electronegativity and electron transfer behavior [4 plasma to fluorinate multi-walled carbon nanotubes and applied it to lithium primary battery and lithium secondary battery and obtained slightly fluorinated multi-walled carbon nanotubes with improved cycling performance compared with the original carbon nanotubes.Many researchers have chosen graphite-phase carbon nitride , Ar/NH3 mixed gas (NH3 concentration: 1.02 × 10−4 mol/mol), and high-purity (>99.996%) CF4 gas were used as the plasma precursor gas; they were purchased from Nanjing Special Gas Co., Ltd. . The used substrates were roving fabrics with the size of 15 mm × 15 mm × 1 mm supplied by Kunshan Zhangsheng Nano Technology Co., Ltd. . The weight of the fabrics with the scale of 10 × 10 mm2 was tested and the values were 77, 81, and 86 mg, respectively, and the average value was 81.3 mg with a standard deviation of 3.68. The used reagent including Ethylene glycol  and acetone  were obtained from Aladdin .The Ar/CHThe roving fabric substrate was washed with acetone solution for 20 min in an ultrasonic cleaning machine to wipe off the surface grease. Then, the fabric surface was washed repeatedly with alcohol and deionized water 3–4 times, and the fabric was dried at 80 °C for 60 min for subsequent experiments.−3 Pa. In this study, different types of films can be obtained by changing the plasma gases, the graphene film was prepared by injecting the 0.25 mL/min plasma gas of Ar/CH4 at 10 Pa of stabilized working pressure for an exposure time of 10 min with RF power of 30 W. After the graphene film deposition, the Ar/CH4 gas was stopped, then the g-C3N4 groups were subsequently generated on the graphene by injecting 0.5 mL/min plasma gas of Ar/NH3 with the discharge power of 100 W at 24 Pa for 120 min, so the g-C3N4/graphene film was produced, and the sample was named as NG. Moreover, the g-C3N4/graphene film was further modified by the CF4 plasma which was generated at the RF power of 100 W under the working pressure of 24 Pa, and the CF4 gas of 0.5 mL/min was injected. Such modification process was performed for 120 min, finally, the CF2 modified g-C3N4/graphene film was prepared, and the sample was designed as FNG. The above abbreviations for the samples refer to the prepared film on the fabric.The synthesis scheme regarding the experimental procedures is shown in −1) and X-ray photoelectron spectroscopy  with the excitation source of a monochromatized Al Kα X-ray source (hν = 1486.68 eV). The binding energies of the XPS peaks were given with an accuracy of ±0.2 eV.Nanostructures and morphologies of the films including graphene, NG, and FNG were tested using high-resolution transmission electron microscopy . Due to the requirement of sample preparation for high-resolution transmission electron microscopy, the film should be stripped from the fabric and evenly dispersed in an alcohol solvent to characterize the morphology and structure of the film. The plasma-treated fabric was placed in an ultrasonic vibrator with an ethanol solution and vibrated for 1 h to ensure that the surface film of the fabric was fully dissolved in an alcohol solvent. The functional groups and chemical bonding types were tested by infrared spectroscopy  in ATR mode (ranging from 4000 to 400 cmWater contact angles (WCAs) of the fabrics covered by the produced films were measured by a contact angle analyzer , and the sessile drop method was used. Stability Testing was performed by an ultrasonic cleaning machine , which provided an important indicator for preparing durable superhydrophobic fabric. The plasma-treated fabric was completely immersed in 95% alcohol and ultrasonic for 20 min, then removed and cleaned with deionized water three times. Finally, the plasma-treated fabric was placed in an oven and dried at 60 °C for 60 min. The contact angle of each sample after ultrasonic cleaning was secondly measured. The average angle was obtained by testing five different locations of each sample with ~10 μL of water droplets. Rolling angles (RAs) were detected using the sessile drop method and 10 μL deionized water droplets with the optical contact angle measuring instrument . RAs were determined by tilting the sample stage at a rate of one degree per second until the drops started moving and rolled off the sample surface.3N4/graphene (NG), and CF2-modified g-C3N4/graphene (FNG) films were generated on the auxiliary electrode for measurements. The Electrochemical impedance spectroscopy (EIS) was obtained by testing the stable open circuit potential of the films immersed in 3.5% NaCl solution at frequencies ranging from 10−2–105 Hz. Additionally, the Gamry Echem Analyst software was used to study the obtained PP and EIS curves.The electrochemical properties of the films were investigated by using the electrochemistry workstation . The pure graphene, g-CThe first principles calculation of the obtained film was performed using the Dmol module in Materials Studio 7.0. The atomic and chemical combination was modified according to the characterization results based on the graphite model after unlocking its symmetry.3N4/graphene (NG), and CF2-modified g-C3N4/graphene (FNG) films. Firstly, the nanostructures of graphene and NG films were captured by TEM, as shown in According to the PECVD deposition steps and different injected gases, three samples of the functional graphene films have been produced, which are the pure graphene, g-C2 modified g-C3N4/graphene film (NG film) sample was detected, and the selected diffraction rings were (010) and (100) of g-C3N4 after the fast Fourier transform (FTT) as shown in Then, the crystal structure of CF2-modified g-C3N4 film (FNG film), the nanostructure and EDS of FNG film were analyzed by TEM, as shown in 3N4 was destroyed due to the modification of the F atom on the film surface, meanwhile, the CF2-modified g-C3N4 presented an amorphous structure, as shown in Zone B. The whole FNG film was scanned by EDS as shown in For the sample of CF3N4 is grown in situ on the basis of graphene which presents a similar crystal structure to graphene, and the nanostructure of g-C3N4 is destroyed after an F atom was introduced on its surface. The FNG film exhibits an amorphous structure on the whole.The above TEM results reflected that all the prepared films are of good quality. For the NG film, g-Cn, CF2, and CF3 appeared at the binding energy of 285.4, 288.4, 289.4, 291.1, and 292.9 eV, respectively [3N4, and CFx originated from the modified CF functional group. The N1s spectrum was fitted to C-NHx, N-(C)3, and N-C=N peaks, whose binding energy values were 401.6, 400.5, and 399.1 eV, respectively [3 and CF2 bonds, where the CF3 bond played the role of structure construction and the CF2 bond endowed the ability of superhydrophobic performance [The survey spectra of all the films were analyzed by XPS as shown in ectively ,38. The formance .x bond are attributed to the graphene structure, g-C3N4, and modified functional group, respectively. The addition of nitrogen and fluorine atoms can enhance the valence bandwidth.From the above results, the composition and functional group types of the film can be obtained. The C-C bond, the N-C=N bond, and the CF3N4 nanostructure with staggered C and N atoms was formed under the premise of ensuring the hexagonal “P1” spatial community of graphene [4 gas and continuous glow discharge, the C-N bond was broken, and an F atom was attached to the end of a C atom to form CF2. Such a process led to serious deformation of the g-C3N4 nanostructure, and C-N close to the F atom shrunk, while C-N far away from the F atom stretched [−1 was carefully scanned. It was found that the C-N absorption peaks can be fitted into three peaks, corresponding to C-N (compress), C-N , and C-N (stretching) [2 functional groups increased the band gap of g-C3N4 from 0.022 eV to 0.217 eV, and the value of the valence band obtained from the density of state was 1.38 by studying the band gap of FNG film [2 functional groups. This phenomenon caused a gap in the electrostatic potential on the graphene surface, which provided a channel for electron transfer, as shown in FNG film was analyzed based on density functional theory (DFT) as shown in graphene . At thistretched , as showetching) ,45, as sFNG film , which c3N4 nanostructure, which created gaps in the electrostatic potential of the graphene surface and provided channels for electron transport, which was consistent with the band gap results of XPS analysis.In combination with the results of infrared analysis, it can be concluded that the introduction of fluorine atoms led to severe deformation of the g-C4 preliminarily treated fabric substrate was hydrophilic with a static contact angle of 79.4°. After the second step of Ar/NH3 treatment, the surface of the treated fabric substrate transformed from a hydrophilic state to a hydrophobic state, with an increased static contact angle of 131.3°. Finally, after CF4 plasma treatment, the fabric substrate possessed superhydrophobic properties with a static contact angle of 156.2°. After ultrasonic cleaning for 20 min, the plasma-treated fabrics attached by the FNG, and NG film maintained favorable hydrophobic performance. Particularly, the superhydrophobic performance of the fabric attached by FNG film was stable on the whole. After ultrasonic cleaning, the fabric attached by FNG film still had a water contact angle as high as 147.6°, which was 8.6° different from the contact angle before ultrasonic cleaning, which indicated that the film exhibited good bonding force with the substrate, and the film exhibited good interlayer bonding force with the film. As for the rolling angles of the samples, the plasma-treated fabrics that were attached by the three kinds of films had similar rolling angles, and the sample with static contact angles over 150° showed a rolling angle of 35.8°.The optical images of the contact angle were obtained by the contact angle measuring instrument, as shown in 2 modified g-C3N4/graphene composite film has a better water contact angle relative to the recent literature review.The superhydrophobic performance of the prepared composite films in this work is compared with the experimental results of other authors, it can be seen in −9, 8.492 × 10−9, 7.603 × 10−9, and 1.059 × 10−8, respectively, which indicated that FNG film still had the ability to decompose reactive oxygen species at a higher polarization current and had good stability [p = Rct + W (where Rct is the charge transfer resistance and W is the infinite diffusion resistance) [The electron transfer of the obtained film was established by an electrochemical workstation, and the polarization curves of the three films and the electrode are shown in tability . The FNGistance) ,55, mean3N4 appeared in the high-frequency region, indicating that FNG film had the characteristics of high pass and the ability of electron transfer [e (ReW), where CPEe is the constant phase angle element of the electrode, Re is the electron transfer resistance of the electrode, and W is the infinite diffusion resistance of graphene, NG, and the FNG film. The Cf and CPEf are capacitance and constant phase angle elements of the film, respectively, Rpo is pore resistance, CPEdl is long phase angle element equivalent to multiple double capacitors, and Rct is electron transfer resistance of the film. The data fitted by electrochemical impedance spectroscopy are shown in 3N4 was alternately arranged by C and N atoms, which formed an uneven three-dimensional structure [The prepared film was analyzed by electrochemical impedance spectroscopy as shown in transfer ,59. Figutransfer ,61, and tructure . The sur3N4/graphene, and CF2-modified g-C3N4/graphene films were synthesized by PECVD. For the g-C3N4/graphene film, g-C3N4 was in-situ grown on the basis of graphene which possessed a similar crystal structure as graphene, while the CF2-modified g-C3N4/graphene film presented an amorphous structure on the whole. Based on density functional theory, first principles calculations combined with infrared experiments revealed that the introduction of fluorine atoms caused serious deformation of the g-C3N4 nanostructure, which created gaps in the electrostatic potential on the graphene surface and provided a channel for electron transfer. The CF2 functional groups promoted the treated fabric to exhibit excellent superhydrophobic properties and stability. The resistance of electron movement of CF2-modified g-C3N4/graphene film was reduced by 2% and 76.7%, respectively, compared with graphene and g-C3N4/graphene. Such sudden reduction of electron motion resistance resulted from longitudinal penetrating equipotential surface caused by introducing F atoms. This method provides ideas for expanding the application scope of multifunctional graphene, but some further studies still need to be conducted to face the problems of film thickness uniformity and large-area deposition for real applications.In summary, by tuning the plasma deposition steps and injecting different plasma gases, the pure graphene, g-C