At present it is not known whether fast [Ca2+]n transients converge in the nucleus with signaling of synapto-nuclear protein messenger. Jacob is a protein that translocate a signalosome from N-methyl-D-aspartate receptors (NMDAR) to the nucleus and that docks this signalosome to the transcription factor CREB. Here we show that the residing time of Jacob in the nucleoplasm strictly correlates with nuclear [Ca2+]n transients elicited by neuronal activity. A steep increase in [Ca2+]n induces instantaneous uncoupling of Jacob from LaminB1 at the nuclear lamina and promotes the association with the transcription factor cAMP-responsive element-binding protein (CREB) in hippocampal neurons. The size of the Jacob pool at the nuclear lamina is controlled by previous activity-dependent nuclear import, and thereby captures the previous history of NMDAR-induced nucleocytoplasmic shuttling. Moreover, the localization of Jacob at the nuclear lamina strongly correlates with synaptic activity and [Ca2+]n waves reflecting ongoing neuronal activity. In consequence, the resulting extension of the nuclear residing time of Jacob amplifies the capacity of the Jacob signalosome to regulate CREB-dependent gene expression and will, thereby, compensate for the relatively small number of molecules reaching the nucleus from individual synapses.Nuclear CaThe online version contains supplementary material available at 10.1007/s00018-023-04876-8. Proximity ligation assays revealed a Ca2+-dependent dissociation of Jacob from LaminB1 which is accompanied by an increased association of Jacob with the transcription factor CREB. Intriguingly, the pattern of nuclear [Ca2+]n transients is strictly correlated with Jacob’s subnuclear distribution. When we induced long-term potentiation with field stimulation in hippocampal primary neurons, we observed an immediate dissociation of Jacob from NL and a subsequent accumulation with the nuclear matrix. Moreover, the dissociation of Jacob from the NL strictly follows the pattern of nuclear [Ca2+]n elicited by different types of synaptic stimulation. We speculate that the Ca2+-dependent association with LaminB1 controls the nuclear function of Jacob inasmuch as its stimulus-dependent availability in the nuclear matrix will contribute to a specific pattern of CREB-dependent gene regulation reflecting different patterns of nuclear [Ca2+]n transients.Here we show that the dissociation of Jacob from the NL strongly correlates with nuclear -4,8-dihydroxyanthracene-9,10-dione (DRAQ5™) was obtained from abcam (ab108410). 4-Aminopyridine  was obtained from Sigma . NMDA , bicuculline methiodide (50 μM), Ifenprodil (5 μM) were purchased from Tocris.The Cat test, nested t test using GraphPad Prism 8 (GraphPad Software). Error bars indicate SEM and sample numbers are either indicated in the graph or in the figure legend. For correlation analysis between Fluo-4 (AM) and Jacob-CyRFP intensity changes, time-lapse recordings were corrected for bleaching and normalized to signal intensities between events. Jacob signal intensities in the nuclear matrix were plotted over Fluo-4 (AM) signal intensities, and a linear regression model and Pearson correlation coefficients were computed to detect correlation.Blind data collection was employed as a protection against bias. Statistical analyses were performed with the paired Student’s We have shown previously that the nuclear import of Jacob critically relies on the activation of GluN2B-containing NMDAR receptors . MoreoveTo address these questions, we first expressed the protein in mature hippocampal neurons, photobleached a small area within the nuclear envelope and plotted the fluorescence recovery after photobleaching .During the fast data acquisition in FRAP experiments, we noticed relatively rapid fluctuations in Jacob fluorescence intensity within the nuclear matrix and at the nuclear rim Fig. c. When w2+-indicator Fluo-4 (AM) and thereby detected local [Ca2+]n. Intriguingly, we found that the time course of Jacob`s redistribution to the nucleoplasm matched the time course of nuclear Ca2+ waves . As expected, we observed a similar pattern of nuclear oscillations for the rise in [Ca2+]n and changes in Jacob-CyRFP2 intensity within the nuclear matrix, suggesting a Ca2+-dependent regulation of Jacob`s subnuclear distribution  boosted Jacob's intensity within the nuclear matrix and, concomitantly, reduced its intensity at the nuclear periphery  Fig. a–c. Trea2+]n transients liberates Jacob from the association with LaminB1, thereby promoting its binding to CREB.We further substantiated these findings in acute hippocampal slices, a physiologically more relevant experimental system with preserved synaptic circuitry. Here we found that in CA1 neurons, endogenous Jacob dissociates from the complex with LaminB1 after brief bath application of bath NMDA Fig. i–l. We hPhosphorylation of Jacob`s serine residue (S180) encodes in the nucleus activation of synaptic GluN2B-containing NMDAR , 54. Nex2+]n transients [2+]n transients n transients and in parallel, the subnuclear distribution of Jacob  to stimulate Schaffer-collateral fibers for induction of LTP in acute hippocampal slices and detected a significant increase in the association of pS180-Jacob with CREB , whose activation is driven in response to Cagh NMDAR . Since agh NMDAR , we obse2+ waves are closely coupled to cytosolic Ca2+ transients, which are in turn coupled to glutamate release that elicits excitatory postsynaptic currents and Ca2+ influx through NMDAR as well as opening of voltage-gated Ca2+ channels and backpropagating action potentials n transients result in a subnuclear redistribution of a synapto-nuclear protein messenger that has a documented role in transcriptional regulation [2+]n concentrations liberate Jacob from its association with LaminB1 and thereby promote its binding to CREB  encodes in the nucleus activation of synaptic GluN2B-containing NMDAR, whereas S180 is not phosphorylated in response to stimulation of extrasynaptic GluN2B-containing NMDAR. The question remains whether Jacob transiently sequestered at the nuclear lamina preserves its phosphorylation status while it is associated with the intermediate filament LaminB1. S180 phosphorylation is crucial for the assembly of a signalosome containing ERK and the intermediate filament α-internexin, which following binding to the basic leucine zipper (bZip) domain of CREB will enhance phosphorylation of S133 and subsequent activation of the transcription factor . WhetherIn contrast to this scenario, following extrasynaptic NMDAR activation, Jacob is not phosphorylated at S180 and it binds preferentially to the nuclear adaptor protein LIM only 4 (LMO4), a transcriptional co-activator of CREB that hinders de-phosphorylation and transcriptional inactivation of CREB . Jacob iIn any of these scenarios, the question arises how long Jacob will stay in the nucleus and how turnover of the protein is regulated since it has to be expected that this is a major regulatory event. A central observation of the present study was that sustained synaptic activity elicited by LTP-like stimulation results in a persistent shift of Jacob from the nuclear lamina to the nuclear matrix. The underlying molecular mechanisms are unclear but because this shift is very likely involved in the regulation of plasticity-relevant gene expression, future studies should aim to learn more about this process. Moreover, the latter observation points to the possibility that in the nucleus basal synaptic, enhanced synaptic and extrasynaptic NMDAR activity can be discriminated by the subnuclear dynamics of Jacob shuttling. Basal synaptic activity results in dissociation and re-association Jacob with the nuclear lamina Fig. . LTP-likIn summary, we here report a novel scenario for activity-dependent gene expression that might have significant functional consequences. The tight control of the dynamics of subnuclear localization clearly deserves more attention in future studies.Supplementary file1 (AVI 7128 KB)Supplementary file2 (AVI 22666 KB)Supplementary file3 (AVI 17428 KB)a Ten-minute representative trace of Jacob-GFP fluorescence within the nuclear matrix of hippocampal neurons under basal glutamatergic transmission. b Depicted are confocal image frames depicting changes in intensities within the nuclear matrix (corresponding to the orange bar in a) (JPG 379 KB)Supplementary file4 Time-lapse imaging of Jacob`s subnuclear redistribution over 10 min. Below is the link to the electronic supplementary material.


Our results illustrate the essential role of SOC in the metal electrode for the CISS spin valve effect. A tunneling model with a magnetochiral modulation of the potential barrier is shown to quantitatively account for the unusual transport behavior.Chirality has been a property of central importance in physics, chemistry and biology for more than a century. Recently, electrons were found to become spin polarized after transmitting through chiral molecules, crystals, and their hybrids. This phenomenon, called chirality-induced spin selectivity (CISS), presents broad application potentials and far-reaching fundamental implications involving intricate interplays among structural chirality, topological states, and electronic spin and orbitals. However, the microscopic picture of how chiral geometry influences electronic spin remains elusive, given the negligible spin-orbit coupling (SOC) in organic molecules. In this work, we address this issue via a direct comparison of magnetoconductance (MC) measurements on magnetic semiconductor-based chiral molecular spin valves with normal metal electrodes of contrasting SOC strengths. The experiment reveals that a heavy-metal electrode provides SOC to convert the orbital polarization induced by the chiral molecular structure to  Chirality induced spin selectivity is a process whereby a chiral molecule induces a spin-polarization to a current passing along the chiral molecule. The exact physical origin of the effect is still debated despite extensive experimental result. Here, Adhikari et al provide evidence for the important role of spin-orbit coupling in the normal metals that connect to the chiral molecule in CISS experiments. More recently, a parallel phenomenon in real space, in which structural chirality induces electron spin polarization in the direction of their momentum, has received increasing attention7. The effect, termed chirality-induced spin selectivity (CISS), was first evidenced by Mott polarimetry of photoelectrons from a nonmagnetic (NM) Au electrode through a self-assembled monolayer (SAM) of short synthetic molecules of dsDNA8. Since then, CISS has been observed in a variety of chiral molecular systems including macro13 and small15 molecules, supramolecular polymers16, metal-organic frameworks17, and hybrid organic-inorganic perovskites19 and artificial superlattices21, via a host of electrical, optical, and electrochemical probes25. More broadly, CISS is shown to effect enantio-selective chemical reactions26 and facilitate enantiomer separation27, and the adsorption of chiral molecules on the surface of a conventional superconductor was reported to induce unconventional superconductivity29. All these experiments suggest a highly consequential interaction between molecular structural chirality and electronic spin, which carries profound and broad implications.Helical textures and monopole-like chirality in electronic structures of topological materials have given rise to a plethora of emergent phenomena characterized by unusual interplays between electronic charge, spin, and orbital31. A central unsettled issue is the role of spin-orbit coupling (SOC) in the chiral media. SOC is a necessary element in the emergence of spin polarization in NM materials in general. Specifically, it is an essential ingredient in most theoretical models of CISS, whereas the SOC in the molecular materials is too weak to account for the experimentally significant spin selectivity at room temperature32. In order to overcome this difficulty, a number of theoretical approaches were proposed, based primarily on spin dependent scattering and tight-binding models41. The approaches have targeted at amplification of SOC, either its value, to account for the experimentally observed CISS spin polarization by introducing various factors such as density of scattering centers36, dephasing42, and environmental nonunitary effects41, or its effect, through electron-electron correlation43, exchange interactions44, vibrational and polaronic effects46, frictional dissipation47, and Berry force48.Despite increasing preponderance of experimental results and a great deal of theoretical efforts, the microscopic origin and physical mechanisms behind CISS remain open questions50. Gersten et al.39 introduced the concept of “induced spin filtering”: A selectivity in the transmission of the electron orbital angular momentum can induce spin selectivity in the transmission process, provided that there is strong SOC in the substrate supporting the chiral SAM. This proposal, however, was questioned because CISS was observed in photoemission experiments in which the substrates have negligible SOC15. Liu et al.49 noted an important difference between the manifestations of the CISS effect in photoemission setups15 and transport in molecular junctions:52 The former measures the “global orbital angular momentum” that includes both the orbital and spin angular momenta, whereas the latter probes spin polarization exclusively. Physically, the model suggests that chiral molecules act as an orbital filter rather than a spin filter, and the SOC in the metal electrode converts the orbital polarization into spin polarization, thus producing CISS without the need for any SOC in the molecules  and MC measurements. Such devices, however, present significant technical challenges of their own: Pinholes are almost always present in SAMs at device scales (~ µm), and in the cases of two metal electrodes, any direct contact will short out the device.One of the most widely used device platforms to detect and utilize spin polarization are spin valves. In a CISS spin valve, the spin polarization of a charge current from the NM electrode through the chiral SAM is analyzed by the magnetic electrode, and the junction conductance is expected to depend on the magnetization direction of the magnetic electrode, resulting in a MC. For CISS studies, a scanning probe rendition of the spin valve, magnetic conductive atomic force microscopy (mc-AFM), has been frequently used52. The use of the magnetic semiconductor was found to alleviate the shorting problem due to the presence of a Schottky barrier at direct contact with the Au electrode. Moreover, the As was grown by molecular beam epitaxy (MBE) on an As buffer layer, and the resulting strain from the lattice mismatch produces perpendicular magnetic anisotropy (PMA)54, namely an out-of-plane magnetization that is collinear with the spin polarization from CISS. These two unique device characteristics enabled observation of spin-valve MC distinctly associated with CISS, and the inherent stability of the platform facilitated a first rigorous determination of the bias current dependence of the MC from CISS52.We recently demonstrated that these complications can be effectively mitigated by using a ferromagnetic semiconductor, As, as the magnetic spin analyzer55 from orbital polarization is shown to provide quantitative account for both the magnitude and bias dependence of the MC of the two types of junctions. The work unambiguously evidenced the essential role of the contact SOC in generating observable CISS effect in chiral molecular spin valves.Leveraging this proven device platform, we fabricated and characterized a deliberately chosen set of As/SAM/NM hetero-junctions. The experiments yielded quantitative differentiation of the magnitude and bias-dependence of the spin valve conductance in junctions with NM electrodes of contrastingly different SOC strengths (Au versus Al) and SAMs of chiral and achiral molecules. The results revealed a definitive correlation between the magnitude of the CISS spin valve conductance and the SOC strength in the NM electrode: The molecular junctions with Au electrodes exhibit significant MC whose magnitudes depend distinctly on the chirality or length of the molecules; in contrast, in otherwise identical devices with Al electrodes, regardless of the molecule involved the MC are essentially indistinguishable from those of the control samples without any molecules. A model based on magnetochiral modulation of the tunneling barrier potential2 to 15 2.We detect the MC in chiral molecular spin valve devices with a As magnetic electrode and Au or Al normal metal electrode. Figure L-polyalanine (AHPA-L) and L-cysteine served to compare chiral molecules of different molecular lengths, whereas 16-mercaptohexadecanoic acid (MHA) and 1-octadecanethiol (ODT) were used as achiral molecules of similar length but with different terminal groups. The total molecular length of AHPA-L is 5.4 nm, whereas L-cysteine is around 4 Å, and those of MHA and ODT are 2.4 nm and 2.7 nm, respectively. All four molecules contain a thiol end group, which facilitates formation of high-quality SAMs on the As57. The experimental details are described in “Methods” section.Molecular assembly, the formation of the SAM on As, is a critical step in the device fabrication process. For this work, alpha-helix 52, despite the probable presence of direct contacts between the NM and As  through defects in the SAM, the spin valve conductance due to CISS in these junctions can be identified from the difference in junction conductance, ΔG, under opposite saturation magnetization for the As. Figure T = 4.8 K. Each MC curve shows two distinct conducting states, coinciding with the well-defined square magnetic hysteresis of the As due to its PMA, hence a ΔG can be precisely determined. As expected, the square hystereses of the MC resemble those of the anomalous Hall effect (AHE) of the As which is directly proportional to M. Any significant contribution of AHE of the As in the junction MC was ruled out in our previous work52. Here we present an updated list of justifications in Supplementary Information (Sec. 5).As described previously52The square hysteresis thus facilitates a straightforward and reliable determination of detailed bias current dependence of ΔG, from I-V curves measured under the opposite magnetization states of the As, as shown in Fig. mentclass2pt{minim52 are presented in Supplementary Information (Sec. 2). We conclude that the total G is spurious (depending the molecular coverage of the SAM) and has no bearing on CISS spin-valve conductance; it is the We note that the total junction conductance (G) for the Au junction is also much greater than that of the Al junction. Nevertheless, measurements on the large number of As/AHPA-L/Au devices clearly demonstrated that although the I-V and total conductance may vary greatly in the molecular junctions of similar structures depending on the degree of parallel conduction , both the magnitude and bias current dependence of the CISS spin valve conductance , each with 4 junctions, and the results were consistent. The experiments, therefore, strongly indicate that the observed significant impact of the NM electrodes on the CISS spin valve effect originates from inherent differences of Au and Al, and is independent of the specific chiral molecule used.Figure We have also fabricated and measured large numbers of devices with Cu and Ag electrodes, two NM materials of intermediate SOC strengths between Au and Al. However, despite the repeated attempts, for either material, we were unable to obtain results with the degree of consistency achieved in Au and Al devices. Most Ag and Cu junctions yielded very small L-cysteine) junctions. Moreover, 49, not necessarily chirality. Both MHA and ODT molecules have a thiol end and different terminal groups, thus are polar and possess inversion asymmetry. Here, the Al junctions again provide an illuminating comparison . It is clear that the MC in the control samples were close to the noise level and lacking any well-defined bias dependence, thus constituting a viable reference for meaningful comparison of the MC in various molecular junctions.We further examined the spin valve effect in the molecular junctions with achiral molecules. Figure son Fig. : There a60, the MC is symmetric As/AHPA-L/Au junction, consistently exhibits significant MC, which increases linearly with the bias current at high biases but approaches a finite value at zero-bias. Contrary to expectation from the Onsager reciprocal relation49 offers a natural account for the observed qualitative differences between the molecular junctions with Au and Al electrodes. More recently, incorporating orbital polarization, a tunneling model was proposed to describe the electronic transport in chiral molecular junctions55. The essential ideas are depicted in Fig. 55. Physically, a current flow induces charge accumulation, which changes the tunneling barrier in a way that depends on the magnetization and molecular chirality. We demonstrate that this model55 can provide a semi-quantitative self-consistent description of all the key observations in the following.The orbital polarization model62 of metal/insulator/metal tunneling junctions. The problem of an arbitrarily shaped potential barrier is modeled into that of a rectangular barrier, which results in an explicit expression for the 64. Based on the Simmons expression and assuming a small magnetochiral modulation of the potential barrier, in the intermediate bias range, we approximate the magnetization-dependent differential conductance through a chiral molecular junction in the form of a simple exponential relation:We incorporate the magnetochiral modulation into the Simmons modelmentclass2pt{minimβ fixed, best fit to the data to Eq. Figure ) to Eq. . More dementclass2pt{minim62 for junctions with dissimilar metallic electrodes and/or asymmetric potential barriers. The fitting results for other junctions, including those of achiral molecules, are described in Sec. 7 of the Supplementary Information. The results provide a quantitative measure of the different effects of the normal metal electrode (Au versus Al) consistent with the qualitative trends of Two notable features are evident in Table entclass1pt{minima55. Physically, the magnetochiral modulation of the tunneling barrier of the insulating molecules can be understood as an analog of the electrical magnetochiral anisotropy (eMChA) in conductors65. It is therefore important to note that eMChA has been observed in both chiral and  polar conductors . Quantitatively, the eMChA is stronger in a chiral crystal with well-defined structural chirality67 than in a nonchiral polar solid. Therefore, finite spin-valve conductance in junctions of achiral (polar) molecules are expected in our model and their magnitudes should be smaller relative to that in chiral molecular junctions. Further theoretical work is needed to relate the quantitative differences with the molecular structures and physical parameters of Au and Al.The observation of CISS MC in the spin valves of the achiral molecules and their magnitudes are well accounted for in our model. The molecular polarity (asymmetry) factors directly into the magnetochiral modulation of the tunneling barrier69. On the other hand, theoretically, there is continuing debate as to whether and how this can be reconciled with the Onsager relation, which precludes a linear-response MC in the two-terminal chiral spin valves71. This topic was a focal point of our previous work52, here we describe a possible pathway out of the dilemma within the proposed model above. Because the model relies on bias current induced magnetochiral modulation of the tunneling barrier, any hysteresis in the charging/discharging process would yield a finite remnant (“linear-response”) spin valve conductance even at zero bias. We note that bias-induced conductance switching, and I-V hysteresis have been reported in a variety of molecular junctions of different configurations and molecules76. Since the conductance is exponentially sensitive to the tunneling barrier, the induced spin valve conductance can be considerable even for a small remnant modulation (splitting) of the potential barrier. In one of the devices we reported on previously , where particular care was taken to examine the effect of bias history, pronounced bias-induced changes in I-V and MC were observed. The results are shown in the current Supporting Information (Sec. 8). In this junction, for applied bias currents up to 800 μA, the junction showed negligible low-bias (linear-response) MC, whereas the application of a 1000 μA bias current induced substantial increases in MC over the full bias range; most notably, a significant zero-bias MC was now present As/AHPA-L/Au junctions. Experimentally, finite low-bias MC has been observed in a variety of two-terminal junctions of different chiral molecules and device structuresion Sec. . In thisIn summary, utilizing a robust device platform of magnetic semiconductor-based molecular junctions proven effective for CISS studies, we have obtained direct experimental evidence that the SOC in the NM electrode is essential to the emergence of the CISS spin valve effect. With a Au electrode, the precipitous decrease of the spin valve conductance from AHPA-L junctions to those of shorter chiral molecule and achiral molecules is readily discerned. Replacing the Au electrode with Al results in pronounced drops of the spin valve conductance for all molecules, to the degree that the differences between the molecules and with the control junctions are no longer discernible. A model based on orbital polarization from inversion-symmetry breaking and the resulting magnetochiral modulation of the tunneling barrier potential is shown to not only consistently account for all key aspects of the experimental results, but also provide resolution to several long-standing open issues in the field, including finite low-bias magnetoresistance due to CISS in two-terminal molecular spin valves, the spin valve conductance being symmetric upon current reversal, and its magnitude often being much greater than what is expected from the spin polarization of the ferromagnet. Our work reveals a close relation between chirality and electronic properties, in which structural chirality information is transferred and transformed from molecular geometry to electronic orbital and eventually to the electronic spin via SOC. The results thus provide useful guidelines for detecting chirality-induced phenomena and designing CISS devices.L-cysteine, MHA, and ODT were acquired from Sigma-Aldrich, Inc. Molecules, except  L-cysteine, were dissolved in pure ethanol at 1 mM concentration. L-cysteine was dissolved in deionized water at a concentration of 2.5 mM. The AHPA-L solution was kept at -18 oC for storage whereas L-cysteine, MHA and ODT solution were stored in ambient conditions.The AHPA-L in the experiments was obtained from RS Synthesis, LLC. The As with perpendicular magnetic anisotropy was grown by low-temperature molecular-beam epitaxy (LT-MBE). 500 nm thick As buffer layer was first grown at 450 °C on semi-insulating (001) GaAs substrates. As films of 40 nm thickness with perpendicular magnetic anisotropy were later grown at substrate temperature of 270 °C. The As thin film has a Curie temperature of 80 K and coercive field of 174 Oe.Junctions Defined by Electron-Beam Lithography (EBL): The sample was spin-coated with 2% PMMA at 4 krpm for 30 s. It was prebaked at 180 °C for 10 min on a hot plate. The EBL was performed to draw the junction patterns of size ranging from 5 2 to 15 2. The sample was developed in methyl isobutyl ketone (MIBK) diluted with isopropanol (1:3) for 40 s and then in pure isopropanol for 30 s at room temperature.Removal of oxide layer on As and molecular assembly: The sample was cleaned with O2 plasma to remove any organic residue. It was set with medium power at 200 mTorr oxygen pressure for 1 min. The sample was then baked at 180 °C for 20 min on the hot plate to harden the PMMA. To remove the native oxide layer on the As, the sample was etched with an ion mill for 1 min 30 sec. It was immersed in ethanol immediately after being taken out from the ion mill chamber before the molecular assembly. For the assembly of molecular monolayer on the As, the sample was left in the molecular solution at room temperature for 24 h. After the assembly, the sample was rinsed with ethanol and dried with nitrogen gas. The molecular assembly process is similar for both chiral and achiral molecules.Top NM Electrodes Deposition: A metallic shadow mask was used to draw the electrodes patterns which was aligned with the molecular junctions in a sample under optical microscope. Thermal evaporation was done to deposit NM electrodes. During the deposition of the top magnetic electrode, the substrate is cooled with liquid nitrogen and the temperature is maintained below –50 oC. 35 nm of Au (without Cr) and 50–70 nm of Al was deposited through a shadow mask.The junction devices are fabricated in the following steps:3He cryostat at the temperature range of 4.2–5.5 K. The measurement procedure is similar to previously reported52. Magnetic field perpendicular to the sample plane was applied up to 700 Oe. The As was first magnetized at 700 Oe, and then the magnetic field was swept at a constant rate of 350 Oe/min for measurements. DC measurements were done with Keithley 2400 as the current source and HP 3458 as the voltmeter. AC measurements were performed with SR2124 dual-phase analog lock-in amplifiers.The sample was fixed on a socket with a copper base and wired with silver paint and Pt wire. All the measurements were performed in an Oxford Supplementary InformationPeer Review FileSource Data


Social media has become an essential channel for the public to create and obtain information during emergencies. As the theme of public concern for emergencies changes over time, there is a lack of research on its dynamic evolution from its latent stage. This paper selects the Henan rainstorm event as a case study and extracts the theme characteristics by combining the life cycle theory and Latent Dirichlet Allocation (LDA) model. It integrates the Term Frequency–Inverse Document Frequency (TF-IDF) and Pointwise Mutual Information (PMI) algorithms as the theme-coding data source to build a dynamic theme propagation model for emergencies. Our research results showed that the theme coding effectively verified the assumption of latent development trends. The dynamic theme model could reveal the theme characteristics of different time series stages of emergencies, analyze the law of the theme evolution of the network’s public opinion, and provide practical and theoretical insights for the emergency management of urban cities. With the continuous development of network technology, tremendous changes have occurred in how information is released, disseminated, and interacted with. As of October 2022, the latest data show that global Internet users have increased by 3.5% in the past year. The global Internet penetration rate has reached 63.5%. In addition, the number of social media users, which takes up 59.3% of the worldwide population, increased by 190 million in the previous year  and can In order to better grasp the golden time after the occurrence of emergencies, gain the initiative, and reduce the severe consequences caused by emergencies, it is imperative to conduct a dynamic evolution analysis on public concern topics on social media platforms under the background of unconventional emergencies, which can provide a theoretical basis for public opinion decision making and analysis. In the literature, researchers divide the whole event into different communication cycles and summarize the public opinion themes in different periods ,6,7 and This paper starts with the latent development time node of unconventional emergencies, takes the recent “Henan rainstorm” urban natural disaster event as an example, analyzes microblog texts in combination with the life cycle theory and hidden Dirichlet model, and explores the topic of dynamic evolution information in the background of the sequence, whereby we mine the evolution law of public attention and put forward suggestions for improvement. Conducting a public opinion evolution analysis can show the discussion hotspots of the network’s public opinion at different stages in the propagation cycle of unconventional emergencies and the dynamic trend changes over time. By counting the number of popular microblogs in the “Henan rainstorm” incident hour by hour, it was found that it took a shorter time to reach the breaking point of public opinion than before, so the advent of the new media era requires that we must plan in the face of public opinion crises. It can help guide public opinion on the Internet when dealing with similar incidents in the future, which will provide a theoretical basis and decision support.With the development of Internet technology, social media has become a popular form of media, and it provides new channels for information sources as well as rapid communication . Social Because social media facilitates the public to publish information in different forms , it has an important place in the field of crisis management . Social In summary, we can understand the important role played by social media in dealing with emergencies, and its rich data provide ideas for diverse research. In this paper, we select data from social media platforms to study the evolution of public attention themes in emergencies and provide theoretical support for emergency management.The “life cycle theory” comes from the evolutionary process of organisms in biology, which change the organisms’ appearance, shape, and function accordingly and show stages. Levitan  argues tThe LDA (Latent Dirichlet Allocation) model is a classic Bayesian model and includes a three-layer structure of documents, topics, and words. It is a probabilistic generation model used to mine potential topics in texts, as shown in The topic model of the LDA model is developed based on the PLSA model. The calculation of the original model is complicated by introducing the Dirichlet prior distribution. LDA is one of the most commonly used text topic mining models. Compared with traditional text semantic analysis, LDA has a better effect in analyzing large-scale unstructured data sets . The datIn the study of opinion topic models, LDA topic models combined with temporal dimensions used to mine the evolution pattern of topic content are commonly used by researchers. Combining other complex models with LDA models to mine text information and change patterns in different dimensions is also a popular research direction. Chen et al.  used theTo sum up, the LDA model is one of the most effective methods used to mine text topics. It has gained significant interest from researchers who are interested in modifying LDA models in order to improve the accuracy of topic mining. However, previous researchers adopted the method of dividing events into different periods and analyzing the characteristics of each stage, which ignores the dynamic evolution process in the development cycle of public opinion events and weakens the connection between different periods. Since there is little research on the incubation period before an emergent event, this paper chooses a recent emergent event with great impact as an example, applies the life cycle theory and the implicit Dirichlet distribution theme model, excavates the theme characteristics of the different stages of public opinion dissemination, and constructs a dynamic theme evolution model through user-defined theme codes to explore the theme dynamic evolution law in the overall process of the event. In order to help deal with similar events, the effective guidance and control of online public opinion provide a theoretical basis and decision support.This study takes the recent unconventional emergency event of “Henan rainstorms” as an example and selects “Sina Microblog” as the data-source platform. In China, microblogs are an important Internet space for public information expression  and are The Henan rainstorm incident was a high-impact event that attracted widespread attention and could thus provide a rich data source for this study. Moreover, regarding the thematic evolution of the incident, it can be found that before the heavy rainfall in Zhengzhou caused significant damage, heavy rainfall extremes had already occurred in neighboring cities with severe consequences. However, because the city is inland, the emergency services personnel lacked experience in managing heavy rainfall disasters, which eventually led to severe consequences. Therefore, the study of heavy rainfall events in Henan Province can provide a reference for other inland cities and a theoretical basis for crisis management intervention points.The initial corpus was built by searching keywords and analyzing the texts of “popular tweets” related to the events. The overall research framework is shown in Based on the division of the evolution stages of online public opinion in the existing literature and the characteristics of the real data set, this paper divides the process of public opinion dissemination into four stages: the incubation period, explosion period, recession period, and stabilization period based on the life cycle theory.First of all, the incubation period refers to the stage when public opinion is just occurring along with the emergence of events, the number of topics related to public opinion is small, the attention is partially great, and the event fails to attract widespread attention.In addition, the explosion period refers to the period when the amount of information related to public opinion grows rapidly and reaches its highest point, the attention to events increases comprehensively, the number of public opinion topics becomes diversified, and the discussion reaches its peak.Furthermore, the recession period refers to the situation where the heat of the matter decreases rapidly after the explosion period. Still, a small-scale rebound of the derived public opinion may occur.Finally, the stabilization period refers to the basic end of the development of the matter and the stabilization of the online public opinion, with the amount of discussion and attention related to the matter showing a decreasing trend.Since the number of topics in the LDA model needs to be set artificially, this paper selected the confusion index to judge whether the selection effect of the number of topics was optimal. Based on the comprehensive performance of the perplexity curve in four periods, the final number of topics was selected as 8. In addition, the selected number of topics achieved a good differentiation effect and did not overfit. The confusion curves for the four periods are shown in The TF-IDF algorithm is commonly used to extract keywords due to its feature of fusing word frequency and word importance. The calculation formula is as follows:PMI  can be used to calculate the similarity between two words. The formula is as follows:One of the most used and influential user interaction platforms in China is the Sina Weibo platform. The microblogging platform was an important channel for public expression and information exchange during the Henan rainstorm outbreak, and it provided a sufficient data volume for this research. In this article, Sina Weibo platform data are used as the data source, and the keyword “Henan rainstorm” was used to explore the popular microblogs from 25 June 2021 to 30 August 2021, and a total of 11,747 data were collected. The data of the Henan rainstorm event were selected from June 25, which is in line with the purpose of mining the latent trend before the event.In order to improve the accuracy of the experimental results, text preprocessing work was performed on all the popular microblogs used, including the steps of jieba word separation, removing deactivated words, and adding user-defined dictionaries. In the process of the experiment, the user-defined dictionary and the deactivated word list were continuously updated and optimized according to the judgment of the model output until the output was relatively satisfactory.The time series distribution of popular microblogs related to the “Henan rainstorm” is shown in The LDA themes were extracted from the corpus of the microblogging texts in the incubation, explosion, recession, and stabilization phases. The final results of the eight topics in each phase and the related feature words of each topic were obtained. In addition, five topic words with high relevance under each topic were selected for theme interpretation. The criteria for selecting highly relevant words were a combination of word probability and topic discrimination in the LDA model results. However, after several experiments, it was found that when only the word probability value was considered when selecting some of the topic features, the top-ranked words in some topics may have been duplicated, which was not conducive to showing the topic differentiation. Therefore, the five words with high relevance that were selected by considering the word probability and topic variability are shown in Themes 5–8 in As mentioned above, the division of the text data topics in the latent period is relatively clear and simple as it is mainly divided into two major parts: “weather warning” and “warm-hearted moments”, and it can be seen that the words related to “rainstorm” appear many times in each division. This can be explained by the fact that the weather changes quickly, and thus the weather forecast information is swiftly updated, which shows that the weather forecast department can effectively warn of possible disasters before the arrival of heavy rain.As shown in As shown in Topic 6 in Topic 7 shown in As shown in To sum up, the text topics in the explosion period were more diversified, with new themes of “help-seeking information” and “relief donation” appearing as the event developed in addition to the themes of “weather warning” and “warm-hearted moment” that already appeared in the incubation period.As shown in The words “assault boat” and “rainstorm mutual aid “ in Topic 3 in As shown in In Topic 7 and Topic 8, as shown in To sum up, although there was no noticeable increase in the theme of the recession period, the content of the theme words was richer. Whether it was “relief donation” or “help-seeking information”, they all showed more diversified information, which was related to the wide dissemination of news reports with a certain lag, which is in line with the law of information dissemination.Many words in Topic 1 in To sum up, the topic of the text in the stabilization period was relatively simple, the development of the event entered a calm state, and the heat surrounding the new round of rainstorm warnings decreased; additionally, by considering all of society’s responses as well as the gradual subsidence of the rainstorm event, the investigation of this rainstorm disaster event was conducted in an orderly manner to give the public an explanation of the event.The LDA model tended to obtain information about the topics contained in the text corpus at a certain stage and the characteristic words within different topics. Some themes may recur in the whole life cycle . But even similar themes had different attention levels in different periods, and their changes could reflect the shift in public attention, which is important when mining the dynamic evolution characteristics of themes.In this paper, the topic hotness value was calculated using topic coding, which is a set of coding rules containing several related feature words. For example, the weather warnings topic contains topic feature words  and the surrounding cities topic contains topic feature words . The encoding rules can be used to capture keywords for the whole life cycle of the text corpus and thus obtain the hotness values of related topics in different periods. This paper integrated TF-IDF keyword extraction and a PMI  algorithm to draw a word co-occurrence network as one of the data sources for constructing topic codes, while the results of the topic words extracted by the LDA model were also used as a supplementary word bank so that the integrity of the topic feature word selection could be effectively realized and hot topic mining and a dynamic topic evolution analysis could be carried out. In order to make a strong distinction between different topic codes, when selecting topic words of different topic codes, one should avoid the selection of possible interference words. For example, the interference word “rainstorm” should not be selected for the topic of help-seeking information.This paper drew a network diagram  that wasThe results of the heat map  and bubbCompared with the incubation period, the topic evolution of the explosion period showed a “diagonal” state, with the proportion of “help-seeking information” and “relief donation” themes increasing rapidly, which was in line with the theme feature word extraction. This indicated that the public released a large amount of unofficial mutual help information through the Internet after the rainstorm and that the rainstorm in Henan attracted widespread attention within a short period of time. The rapid increase in the proportion of the “relief donation” theme showed that the unity in the blood of the nation is stimulated in the face of sudden disasters, and the whole nation was united to help Henan.Compared with the explosion period, the proportion of “help-seeking information” in the text topics in the recession period decreased, and the keywords in the user-defined dictionary were mainly divided into two aspects: real-time help information and postdisaster urgent supplies. In the explosion period, the proportion of real-time help information was larger, while in the recession period, the theme of “help-seeking information” mainly focused on the information of the materials needed after the disaster. The proportion of “warm-hearted moments” kept increasing and reached the top value in the recession period, which was consistent with the extracted results of the theme words due to the repeated nature of the rainstorm and the delay of news reporting.After entering the stable period, it can be seen that except for “Safety tips”, all the other topics were rapidly declining in popularity, and the development of the event entered a stable stage with intermittent small climaxes due to the repetitive nature of the heavy rainfall. The combination of “weather warning” and “safety tips” showed that the public became more alert to the new round of rainfall after the heavy rainfall in Zhengzhou, and all relevant departments were fully prepared to face the possible situation. Relevant departments should pay timely attention to the impact of the rainstorm disaster on all walks of life. For example, as the event entered a stable period at the end of August, schools faced the opening of schoolwork, postdisaster reconstruction, the resumption of work and production, and other issues, which required the introduction of relevant measures.In summary, the user-defined topic codes could clearly show the changes in the proportion of different topics in the whole life cycle of event development, tap the patterns between different periods, and better show the hot trends of discussions at each stage of the public opinion dissemination cycle in the context of sudden natural disaster events, which can help relevant management departments give early warning tips when dealing with similar events in the future and can provide a theoretical basis and decision support to guide online public opinions.In the era of rapid Internet development, information breaks the traditional channels of information dissemination with the help of social media platforms, which revolutionize different aspects of its generation, dissemination, and reception . Social For the government, the timely handling of relevant public opinion is the best solution. After the occurrence of an emergency event, the government should be the first to promptly release the progress of the event through an official platform, constantly update the information, carry out the investigation and analysis of the event, update the causes of the emergency event and the progress of the investigation, and listen to the public opinion to ensure that the dissemination of information is not one way . The pubNowadays, posting on social media platforms is the way that companies promote their brands, products, and corporate images ; thus, tThe media plays a vital role as a link between the government and the public, and the timeliness of news is its value. In the face of emergencies, the network will be flooded with a large number of reports on emergencies. The news media should make a rigorous and authentic report on the event itself and continue to follow up on the event’s progress. The news report itself enables the public to more easily understand the event through the Internet, so the news report should uphold a true and rigorous attitude. All information should be based on the actual situation of the event. Therefore, news reports should keep a thorough attitude, and all information should be based on the actual development of the event; eye-catching news reports that simply draw attention should not be written. Fake news is a burden on contemporary society and has a significant impact on citizens, countries, and organizations .In particular, fragmented reading has become a popular way to occupy our lives in the age of self-media. Short videos and articles can attract a lot of attention through eye-catching titles, but such a format may be accompanied by the covert spread of online rumors. The public interprets things through fragmented texts and videos and adds subjective understandings and emotions for secondary dissemination, which gives favorable conditions for the fermentation of online rumors. Therefore, news media should be rigorous in reporting based on facts.From the empirical analysis of theme mining, it can be seen that the theme of the needs of the public after an emergency is divided into different modules. During the rainstorm in Henan, rainstorm mutual assistance became one of the crucial unofficial help platforms for the public. Through the explosive release and reprint of information on social media platforms, there will inevitably be a duplication of information and failure to update the rescued information promptly, which will result in a waste of rescue efforts. In media platforms, this process should be improved from the perspective of user needs. Social media platforms can establish an official topic channel of the media to summarize information and update it promptly to avoid the resource waste caused by information redundancy.By analyzing the dynamic evolution process of the event theme, we could mine the change rule of public opinion development and the change in hot topics in different periods from the complex data, which could thus help the relevant management departments improve their efficiency and decision support when dealing with similar events in the future.The rainstorm event in Henan that was selected for this study was of practical research significance. The rainstorm event in Henan reflected the lack of experience of inland cities in natural disaster emergency management. Ecological and environmental issues are the key concerns of today’s society, and we should learn from the experience of the Henan rainstorm event to provide ideas for the emergency management of other inland cities. The dynamic evolution process of theme mining and the theme hotness value can provide theoretical support for relevant managers to understand public needs and grasp the changes in public needs to improve emergency relief work. The multiperspective discussions among enterprises, media, and the government can enable different participants to gain experience and improve their work. The theme of “surrounding cities” was used to verify the assumption of the existence of latent phase crises and to enrich emergency management research. By combining the life-cycle theory and the LDA theme modeling, this study mined and analyzed the themes of texts at different stages of the communication cycle and explored the dynamic evolution of themes in the development of public opinion by setting user-defined theme codes and mining the hot themes at each stage. In terms of theory, this study combined the LDA theme mining model and life cycle theory to realize the integration of theory into practice. Previous studies have mainly analyzed each stage of public opinion dissemination independently. Still, this paper established user-defined topic codes based on the whole corpus collected. It reflected the dynamic evolution of various topics in the life cycle of public opinion dissemination more accurately and intuitively through visualization results, which provided relevant ideas for public opinion topic mining and evolution analysis.In future research, we will consider collecting data from various platforms, such as the popular short-video platform TikTok, where videos give users a richer sensory experience. Still, the update of communication media will also lead to an increase in the risk of emotional public opinions on emergencies. Meanwhile, we need to enrich the selection of breaking news cases to improve the practical significance of our findings. We will break the research boundaries of different influencing factors by constructing an opinion network and linking user nodes with themes and emotional factors to study the impact of multidimensional online public opinions.


The ejaculatory reflex comprises a complex reflex synchrony involving the central and peripheral nervous systems. Receptors and sensory areas, afferent pathways, cerebral sensory areas, cerebral motor centers, spinal motor centers, and efferent motor centers are involved in the ejaculatory reflex. These areas also jointly modulate other aspects of sexual response .Neurochemically, this reflex involves a complex interaction between serotonergic and dopaminergic and, secondarily, cholinergic, adrenergic, oxytocinergic, and γ-aminobutyric (GABA) neurons. While dopamine promotes seminal emission/ejaculation via D2 receptors, serotonin is inhibitory. This aspect needs to be clear, as knowledge of this neurophysiology will guide the treatment: serotonergic stimulation of 5-HT2c receptors results in delayed ejaculation, while stimulation of postsynaptic 5-HT1a receptors results in shortening of ejaculatory latency .The peripheral events leading to ejaculation are controlled by the synergistic activation of autonomic functions (sympathetic and parasympathetic) and somatic portions of the nervous system. The origin of the autonomic and somatic motor efferent roots originates in the thoracolumbar and lumbosacral segments. Autonomic and somatic coordination are controlled in the L3–L5 region by a group of neurons called spinal ejaculatory generators. This nucleus, in conjunction with the autonomic system and the spinal somatic nuclei, is capable of stimulating or inhibiting the entry of genital and supraspinal sensory stimuli.In higher centers, specific areas are activated during ejaculation, such as the bed nucleus of the stria terminalis, the amygdala, the preoptic nucleus, and the thalamus.We prepared a didactic scheme that summarizes the aforementioned aspects:After this review of neurophysiology, we can move on to deepening the ejaculatory process. To facilitate understanding, it is subdivided into three synchronized phases: emission, ejection (or penile expulsion) and orgasm.Emission consists of contractions of the vas deferens and seminal vesicles, with the expulsion of sperm and seminal fluid into the prostatic urethra. This step is mediated by synergistic sympathetic and parasympathetic activation (T10-L2). The ejection phase is mediated by a spinal sympathetic reflex and somatic nerves (S2 to S4) from the pudendal nerve and involves pulsatile contractions of the bulbospongiosus, ischiocavernosus, and levator ani muscles along with relaxation of the external urinary sphincter. The internal urinary sphincter (bladder neck) closes to prevent retrograde flow into the bladder. The external urinary sphincter also presents cyclic contractions during semen ejection, helping in its expulsion.Ultimately, orgasm is the result of brain processing of sensory stimuli from the pudendal nerve by increased pressure in the posterior urethra, sensory stimuli from the verumontanum, and contraction of the bulbourethral and sexual organs.These are a class of drugs prescribed for the treatment of a variety of mood disorders, such as depression. O use of SSRIs to treat PE is based on the observation that delayed ejaculation and anorgasmia are common side effects of this class of drugs used for a long time off-label. SSRIs, alone at low doses or in combination with psychosexual counseling, are widely accepted as first-line treatments for lifelong PE and are recommended by leading urology guidelines.Men with acquired PE usually receive targeted therapy with the aim of resolving the underlying etiology of your PE, with or without the addition of SSRIs, depending on clinical judgment. SSRIs act to block the axonal reuptake of serotonin from the synaptic cleft of central serotonergic neurons by 5-HT transporters, which desensitize 5-HT1A and 5-HT1B receptors  5, 6)..6).Although ejaculation delay can be observed within a few days, it is necessary to administer the drugs for at least 2-3 weeks to maximize their therapeutic effects. Most SSRIs have been shown to delay ejaculatory time by increasing the average IELT by up to 13.2 times. However, chronic use of these drugs can increase the likelihood of unwanted adverse events such as decreased libido, anorgasmia and impotence/erectile dysfunction, and their sudden discontinuation can lead to SSRI withdrawal syndrome, which can last for more than a week and be accompanied even by suicidal ideation. Therefore, caution is required in handling these medications, starting use gradually as well as gradually weaning. Following this reasoning, the ideal SSRI should have a rapid onset of action and elimination, good tolerability, fewer adverse effects and be formulated for use as an on-demand treatment. And here we come to the high point of this editorial: dapoxetine is a short-acting SSRI option that meets the above PE treatment requirements.Dapoxetine, or dapoxetine hydrochloride (N-dimethyl-(a)-[2-ethyl]-benzenemethanamine) has a structure similar to fluoxetine and works similarly to other SSRIs, inhibiting the transporter that performs serotonin reuptake in the pre-synaptic cleft, whith minimal effects on norepinephrine and dopamine reuptake receptors. It is rapidly absorbed after administration but may change absorption in the presence of other foods. Its elimination is biphasic, with an initial half-life of approximately 1.3-1.4 hours and a terminal half-life of 21 hours, depending on the dose. Compared to longer-acting SSRIs such as fluoxetine and paroxetine, dapoxetine reaches steady-state plasma concentrations within four days due to its short half-life, being metabolized in the liver by cytochrome P450 isoenzymes and excreted in the urine without any effects on the enzymes of cytochrome P450 .It is worth mentioning that for patients who present an anxious component, dapoxetine used on demand will not treat generalized anxiety, and its indication should be made in accordance with the observation of this clinical aspect. For this profile of patients, exclusive use on demand will not bring benefits to other areas of life, and the very use of dapoxetine with the aim of improvement in the sexual sphere, due to the basic anxious character, can induce a state of anxiety regarding an expected response in the improvement of the ejaculatory pattern.It is also important to point out that in the case of choosing to replace a long-acting medication (SSRI) with dapoxetine, adequate weaning over the course of weeks should be carried out, not simply replacing the drugs.Finally, as a conclusion, we observed that dapoxetine is a drug that fills an important space in the management of premature ejaculation, offering a safer and more comfortable treatment, mainly for the urologist, through the administration of an on-demand central nervous system action drug.


This has exerted a severe environmental impact, so in our work, we propose the utilization of a sustainable electrospun nanofiber based on poly lactic acid (PLA), as it is biobased and conditionally degradable. We optimized the weight percentage of the PLA-precursor solution and found that 19% PLA produces fine nanofibers with good morphology. We also introduced carbon nanodots (CNDs) in the nanofibers and evaluated their antibacterial efficiency. We used 1, 2, 3, and 4% CNDs with 19% PLA and found increased antibacterial activity with increased concentrations of CNDs. Additionally, we also applied a spunbond-nanofiber layered assembly for the medical face masks and found that with the addition of only 0.45 mg/cm The sudden outbreak and rapid transmission of the SARS-CoV-2 COVID-19) virus has encouraged people across the globe to adopt safety measures to help stop the spread of this infectious disease. Thus, there has also been a sudden increase in the demand for PPE, such as face masks and gowns. Globally, the market size of PPE in 2022 was USD 80 billion, and this was projected to rise to USD 111 billion by 2029. Furthermore, due to COVID-19, the market size for PPE was increased by 17% in 2020 compared to 2019 9 virus h. Its firThe transmission of COVID-19 is mainly caused by aerosols exhaled by a COVID-positive individual to a by-stander within a range of 1 m. People infected with COVID-19 can spread the virus in the shape of respiratory droplets 5 microns in size when they exhale, cough, sneeze, speak, or sing. These exhaled respiratory droplets (containing the COVID-19 virus) can be inhaled by other people directly and can also be transmitted if the droplets touch the eyes of other people . Another2 and found that the bacterial filtration efficiency was above 97.9%. Choi et al. [As the sudden outbreak has increased the usage of disposable medical face masks, globally, this has generated an estimated waste of 7200 tons per day . Generali et al.  developei et al.  worked oi et al.  printed i et al.  developei et al.  proposedE. coli cells both in suspensions and on the agar surface.Whether by using antimicrobial polymers or by adding antimicrobial agents, researchers have opted to impart antimicrobial properties to the polymers used for medical face masks. In our previous study, we also studied the comparative antimicrobial properties of Aloe vera and ZnO NPs in PVA nanowebs. We found that the ZnO NPs offered superior antimicrobial activity to Aloe vera, which was entirely due to the photocatalysis property of ZnO NPs. Among the emerging material with antimicrobial properties are carbon nanodots (CNDs), or carbon quantum dots . These CS. aureus and E. coli. To our knowledge, this work has not yet been reported.To address the current environmental impact and disposal-related issues with medical face masks, we propose to develop a nanofiber sheet or web to replace the inner layer in medical face masks. The inner layer in medical face masks is composed of a melt-blown nonwoven web, and it essentially serves as a filtration layer, sandwiched between two layers of spunbonded nonwoven sheets. We also propose that the material cost is reduced to almost 75% with the nanofiber layer compared to the meltblown layer. Furthermore, we introduce CNDs as emerging photoactive materials in PLA nanowebs, having found that they offer good antimicrobial properties against The biopolymer polylactic acid 4060D, in pellet form, was purchased from NatureWorks LLC. It has a melting temperature of 210 °C. Dimethyl formamide was purchased from Merck. Glucosamine hydrochloride (98% purity) from Thermos Fischer, Shanghai, China and 1,3-diaminobenzene (99% purity) was purchased from Carl Roth.2O (20 mL) and agitated to achieve complete dissolution in 250-mL Erlenmeyer conical flask. Next, 1,3-diaminobenzene  dissolved by sonication in methanol (10 mL) was added to the glucosamine solution and further agitated to obtain solution homogeneity. The conical flask was then placed in microwave, in a well-ventilated area, and then irradiated for 3 min . A viscous brown oil-like material was produced, which was dissolved in water (10 mL) and centrifuged at 4000 rpm for 1 h. Next, the bulk solution was reduced in vacuo to obtain brown powder. The CNDs were then dispersed in DMF and used in different weight percentages following the design of experiment defined in For the synthesis of CNDs, glucosamine hydrochloride  was dissolved in deionized HThe PLA was first dissolved in DMF in desired quantity and then stirred for about 6 h to produce a homogenous solution. 2 and the feed rate was maintained through a syringe pump at 0.4 mL/h. The machine was enclosed in a see-through container and temperature/humidity were maintained automatically by a device. The needle was placed vertically to that of collector drum with a maintained distance of 17 cm. A potential difference of 23 KV was applied for the generation of PLA nanofibers. The nanofiber sheets were prepared by electrospinning technique. All solutions with varying amounts of PLA and CNDs are presented in The rheological properties, such as viscosity of PLA solutions, were measured by Viscometer B-ONE PLUS (LAMY RHEOLOGY Instruments). First, a zero setting was measured by removing the spindle, after which viscosity of PLA solution was measured with spindle R-6, at 600 rpm, for 60 s. The electrical conductivity of PLA solutions was determined with conductivity meter . The surface morphology of nanofibers was observed with scanning-electron microscope (SEM), . To check the fiber diameter, we analyzed the SEM images with ImageJ software by taking an average of thirty readings from each sample. Dynamic light scattering (DLS) was performed to determine the size distribution of the CNDs using a Zetasizer 1000 apparatus . The sample was diluted to minimize the agglomeration. Measurements were conducted at an angle of 90° and a temperature of 25 °C. To check the chemical composition of the formed nanofibers, we subjected them to FTIR (Shimadzu IRPrestige21) spectroscopy. Thermogravimetric analysis (TGA) (TA Instrument TGA55) was performed to check any residual DMF in the nanofibers. The aluminum oxide crucible was used to perform TGA with a heating rate of 10 °C/min.E. coli) and Gram-positive bacteria (S. aureus). The test specimens were taken in a specific grammage and then placed in the vial. As the samples were hydrophobic, they were placed in the bacterial solution with the help of glass rod. All vials were shaken well on a mixer to obtain homogenous solution. All vials were then incubated with a D-65 light source at 37 ± 2 °C for 18 to 24 h. After incubation, the antimicrobial activity was calculated according to ISO 20645: 2005 Annex C: Quantitative measurement by plate-count method. The antimicrobial activity was measured against the control vial with pure PLA nanofibers.The antimicrobial activity was analyzed by plate-count method (quantitative measurement), defined in ISO 20645:2004 standard. The PLA/CND electrospun nanofibers were tested against Gram-negative bacteria  to that in the filtrate (downstream). The PFE utilizes filtered and dry air through an automizer, which is used to produce aerosol containing latex spheres with particle sizes in range of 0.1 to 5.0 μm and airflow-test velocities of 0.5 to 25 cm/s. Each sample was clamped into the fixtures and then mounted on the machine, and the test procedure was initiated. The generated aerosol was passed through the sample material, and the photometer detected the upstream and downstream counts of aerosol particles and generated a report on percentage filtration efficiency for different sizes of aerosol.2 and differential-pressure-sensor range from 50 to 500 Pascals. Following EN 14683 test method, air was drawn at the rate of 8 L/min to a measured area  of the sample, and the differential pressure was measured in Pa/cm2.Air permeability of prepared samples was determined by a differential-pressure tester from Qinsun Instruments Co., Ltd.,  model G285, with test head of 4.9 cmThe SEM analysis described the morphology of the as-formed nanofibers on the nonwoven sheets. With the help of ImageJ software, we measured the average diameter of each sample. About thirty measurements were taken on each SEM image on the same fiber at different locations and on different fibers, and then the average diameter of the nanofibers was calculated. One of the most important parameters that influences the uniform formation of electrospun nanofibers, and their diameter is the concentration of the precursor solution. We started with 12% pure PLAs in DMF (without CNDs) to check their spinnability, and we observed that rather than forming nanofibers, the droplets were collected on the drum collector. This was because low concentrations provide low viscosity, which favors droplet formation rather than nanofiber formation . Figure For the electrospinning of the PLA/CNDs, it was observed that with the increase in the concentration of conductive CNDs, the obtained average diameter of the nanofibers decreased, as shown in Similar behavior was also observed in the samples with 19% PLA with varying concentrations of CNDs, as shown in The hydrodynamic size analysis was conducted on a zeta sizer (DLS). S. aureus or the E. coli. In contrast, the integration of the CNDs resulted in an antimicrobial activity and with the increase in the concentration of the CNDs in the PLA nanofibers, the antimicrobial activity also increased.Amongst all the weight percentages of the PLA in the DMF, we found that 19% PLA produced fine nanofibers. Therefore, we tested the antimicrobial activity of the PLA-19 with different concentrations of CNDs. E. coli are shown in E. coli, PLA-19/C1 exhibited only 10% antimicrobial efficiency, which increased to 62.5% with the 4% CNDs. Since the CNDs were also positioned on the surfaces of the PLA nanofibers, more sites for reactive oxygen species were present and, in turn, more microbes were killed.The quantitative measurements of the antimicrobial activity against S. aureus are shown in S. aures as compared to the E. coli. This can be explained by the fact that the cell wall of the Gram-negative bacterium, E. coli, is sandwiched and protected by two phospholipid bilayers; thus, the outer lipid layer resists the reactive oxygen species, resulting in less antimicrobial activity [The quantitative measurements of the antimicrobial activity against the activity ,26.The PLA/CNDs nanofibers were electrospun on a PLA nonwoven fabric. After 24 h of electrospinning, we performed the FTIR analysis. Due to the reduced electrospinning time, the weight percentage of the nanofibers collected on the nonwoven PLA sheet was very low. Due to the stacking of the FTIR data of all the samples and comparing them to the nonwoven PLA-collecting sheet, no differences were present in the samples, as shown in The as-collected nanofibers on the nonwoven sheet were then subjected to thermogravimetric analysis (TGA), which is an analytical technique used to determine the thermal stability of materials and its fraction of volatile components by monitoring the weight change that occurs as a sample is heated. The samples were subjected to TGA analysis after 24 h of formation. The TGA curve in As the main attribute of a medical face mask is its filtration efficiency, we conducted the particle-filtration-efficiency and differential-pressure tests, which are the main requirements according to ASTM-F2100-20. We performed the electrospinning of PLA-19/C4, as it showed the highest antimicrobial efficiency amongst other concentrations. This PLA-19/C4 was then electrospun on the nonwoven sheet for different time periods 30, 60, 90, and 120 min.2, respectively. These values are perfect for Type I medical face masks, as mentioned in ASTM-F2100-20.2) compared to the meltblown sheet (1.77 mg/cm2), which is normally used as a filtration layer in medical face masks. Thus, by using sustainable, biodegradable PLA/CND nanofibers as a filtration layer, we saved almost 75% of the PLA material’s cost without compromising the barrier performance of the medical face mask.With respect to the different electrospinning times, the weight add-on to the rotating-drum collector is shown in 2. We suggest that the use of this nanofiber layer sandwiched between spunbond layers makes a perfect filter layer that can be used for Type I medical face masks. The PLA is also a biobased and conditionally degradable polymer, which can also address the waste problems incurred by medical face masks made from non-sustainable sources.This study concludes that the use of 19% PLA in DMFs produced nanofibers with perfect morphology. The other concentrations of 12, 15, and 17% produced nanofibers but with more beads, and we also found that with the increase in concentration from 12% to 19%, the bead formation decreased and finer, smoother nanofibers were formed. As carbon nanodots are reported to have photocatalytic properties, we introduced different proportions of CNDs in PLA-19 nanofibers and found that with the increase in the concentrations of the CNDs from 1 to 4%, the antibacterial efficiency was also increased. In addition, we found that when the PLA/19-C4 was electrospun for 90 min on a spunbond sheet, it formed a perfect layered assembly, with a good PFE, of 96.5%, and a differential pressure of 39 Pa/cm


Pathological calcifications may consist of calcium oxalate (CaOx), hydroxyapatite (HAP), and brushite (BRU). The objective of this study was to evaluate the effect of phytate , InsP6 hydrolysates, and individual lower InsPs  on the crystallization of CaOx, HAP and BRU in artificial urine. All of the lower InsPs seem to inhibit the crystallization of calcium salts in biological fluids, although our in vitro results showed that InsP6 and InsP5 were stronger inhibitors of CaOx crystallization, and InsP5 and InsP4 were stronger inhibitors of BRU crystallization. For the specific in vitro experimental conditions we examined, the InsPs had very weak effects on HAP crystallization, although it is likely that a different mechanism is responsible for HAP crystallization in vivo. For example, calciprotein particles seem to have an important role in the formation of cardiovascular calcifications in vivo. The experimental conditions that we examined partially reproduced the in vivo conditions of CaOx and BRU crystallization, but not the in vivo conditions of HAP crystallization. Phytate was first discovered in vegetable seeds during the 1850s, its chemical formula  was established in the early 1900s, and the first animal experiments assessed the effects of phytate consumption in the 1940s . These sThe activity of phytate and its derivatives as inhibitors of the crystallization of calcium salts must be attributed to the presence of various phosphate groups in its molecule, which can interact with areas of nuclei or crystalline surfaces where calcium is found. In fact, all polyphosphates have the capacity to inhibit the crystallization of calcium salts, with pyrophosphate being the first to be described . PreciseA notable difficulty in studies that evaluate the effects of InsP6 and its metabolites is determining the levels of all these substances in biological fluids and tissues . One issPyrophosphate is one of the first described inhibitors of calcium salt crystallization in living organisms ,19. The The objective of this study is to evaluate the effect of InsP6, mixtures of InsP6 hydrolysates, and individual lower InsPs  on the crystallization of three compounds that are fundamentally involved in pathological calcifications: calcium oxalate (CaOx), hydroxyapatite (HAP), and brushite (BRU).The synthetic urine components were obtained from PanReac . Myo-Inositol-1,2,3,5,6-pentaphosphate , myo-Inositol-2,3,5,6-tetraphosphate , myo-Inositol-1,4,5-triphosphate , and myo-Inositol-2,4-diphosphate  isomers were purchased from SiChem . Phytic acid sodium salt hydrate and etidronate were purchased from Sigma-Aldrich . Bisphosphonates, alendronate, pamidronate and ibandronate were purchased from Molekula . All solutions were prepared in ultra-pure deionized water from a Milli-Q system.A phytate stock solution (2 mM) was prepared from phytic acid sodium salt and adjusted to pH 2 using 0.5 M HCl. Duplicate aliquots (5 mL) were kept in a metallic dry bath block heater  at 97 °C for 6, 9, 16, 24, 48, or 72 h to obtain the mixtures of its dephosphorilation products at different hydrolysis time.The estimation of the evolution of the relative proportion of the InsPs of hydrolysates during thermal hydrolysis was performed by MS/MS spectrometry and the concentration of inorganic phosphate released was determined.m/z and a resolution of 140,000.A Q Exactive Orbitrap high-resolution mass spectrometer equipped with a heated electrospray ionization (HESI) probe , operated in a negative ionization mode, was used to evaluate the evolution of the relative proportion of the InsPs in function of the hydrolysis time. The temperature of the ion transfer capillary was set to 320 °C, the spray voltage was set to 2.9 kV in negative mode, and the S-lens RF level was 50 AU. Direct injection of properly diluted samples was performed in the full-scan acquisition mode over a range of 150 to 700 The amount of inorganic phosphate that was liberated during InsP6 hydrolysis was determined using ammonium molybdate and ascorbic acid (a reducing agent), according to the phosphomolybdate–ascorbic acid method , measuriThe consideration of the relative MS/MS peaks of each InsP at the different hydrolysis times together with the corresponding liberated inorganic phosphate concentrations allows the estimation of the evolution of the concentration of each InsP in the hydrolysate samples.The effects of phytate and the mixtures of its dephosphorilation products at different hydrolysis times on the crystallization of CaOx, HAP, and BRU in synthetic urine were assessed using a kinetic turbidimetric system, as previously described . This syi) was set as the point at which the absorbance cero line crosses the tangent line of the first straight zone of the absorbance curve vs. time. The effects of tested substances on the induction time for the crystallization of CaOx, BRU, and HAP were expressed as the increment of that time  with respect to the induction time of the corresponding control, that is, Δti = induction time − induction time of control.The induction time  was added to induce CaOx crystallization.For the BRU and HAP experiments, 100 mL of synthetic urine solution A at pH 6.5 or 7.5, respectively, was transferred into a 250 mL crystallization flask placed in a thermostatic bath at 37 °C, and 0.2 mL of InsP6 stock solution or a hydrolyzed mixture was added. When the resulting solution reached a temperature of 37 °C, 100 mL of synthetic urine solution B at 37 °C and pH 6.5 or 7.5, respectively, was added to induce crystallization. All experiments were performed in triplicate.The morphological and structural characteristics of the CaOx, BRU, and HAP crystals that formed in synthetic urine in the absence or presence of different InsPs were examined using a scanning electron microscopy system  coupled with XR energy dispersive microanalysis . Crystals formed during crystallization experiments in the absence and presence of additives were collected at the end of each experiment by passing the solution through a 0.45 µm filter. They were then dried in a desiccator and examined by SEM. The methodology used for SEM consists of placing the crystals on a sample holder with fixation on adhesive conductive copper tape, with no need to cover the sample with gold. FTIR spectra of crystals were also obtained by the KBr pellet method with a Bruker Hyperion IR spectrometer  to confirm the crystalline phases identified by SEM.We first recorded the evolution of hydrolysates (dephosphorylation products) of InsP6 during thermal hydrolysis in a metallic dry bath block heater at pH 2.0 and 97 °C . The InsWe then determined the effect of InsP6 and the mixtures of its dephosphorilation products at different hydrolysis times on the time needed for CaOx crystallization in artificial urine at pH 6.0 and 37 °C . In thisWe then compared the effects of different specific InsPs on the induction time of CaOx crystallization . The resWe also compared the effect of four different bisphosphonates at the same concentration to InsPs (2 µM) on the induction time for CaOx crystallization. Under the conditions studied, alendronate, pamidronate, ibandronate, and etidronate had much weaker inhibitory effects  than InsP6 and its hydrolysates (up to 18 min).As can be seen in We used the same basic procedures to determine the effect of different InsPs and bisphosphonates on the formation of BRU crystals in artificial urine at pH 6.5 and 37 °C. The results showed that the InsP6 hydrolysates collected at 16 h and 24 h (which were abundant in InsP4 and InsP5) had much stronger inhibitory effects on the crystallization of BRU than InsP6 alone . HoweverWe then examined the effects of four different bisphosphonates (2 µM) on the inhibition of BRU crystallization relative to 2 µM phytate alone . AlendroWe also determined the effects of four different bisphosphonates and InsP6 and its hydrolysates on the induction time of HAP crystallization . These rWe performed scanning electron microscopy to confirm the inhibitory effects of the different treatments. The results showed that CaOx crystallized as a CaOx trihydrate A in the The present study extended our previous findings  by examiAs shown in previous studies ,23, thesOur study on InsP6 hydrolysates suggested that InsP4 inhibited CaOx crystallization, but the effect of the pure 2,3,5,6-InsP4 isomer was less than expected. This might be because 2,3,5,6-InsP4 is a weaker inhibitor than the InsP4 isomers that are present in the hydrolysate mixture. Alternatively, it is possible that there are synergistic effects in the mixture of InsP hydrolysates. As noted above, the plasma and urinary levels of InsP4, InsP3, and InsP2 are also much higher than those of InsP6 and InsP5 ,23, and Our examination of BRU crystallization showed that the 16 h and 24 h hydrolysates (which were enriched with InsP5 and InsP4) had the strongest effects. Nevertheless, our examination of individual InsPs showed that InsP6, InsP5, and InsP4 had similar effects. As outlined above, these results can be explained by synergistic effects of the multiple InsPs or different InsPs isomers in the hydrolysate.Interestingly, we found that InsP6 and its hydrolysates had little effect on the crystallization of HAP, and that bisphosphonates (except ibandronate) also had little effect. In contrast, studies with experimental animals and humans showed that the intake of InsP6 significantly reduced HAP calcification ,25,26. TOur results suggest that the effects of InsP6 on experimental animals and humans on crystallization are a consequence of the mixture of the different InsPs. These mixtures of hydrolysates can form in the intestine prior to paracellular absorption (despite the absence of phytases in the human intestine) because certain foods contain phytases. Once in the blood, phytases/phosphatases produced by humans and experimental animals can give rise to lower InsPs. Obviously, we did not demonstrate whether the in vivo effects are due to the presence of certain isomers, differences in the concentrations of different InsPs, or interactions among different InsPs. To answer these questions, it is necessary to use analytical methods that can determine all of these different InsP isomers in biological fluids, and appropriate InsPs standards must be available. At present, most of these standards are not available, and the currently available analytical methods do not allow the precise determination of different InsPs isomers.Finally, it is necessary to consider that in addition to the direct interference of crystallization by the different InsPs examined here, many other dephosphorylated InsP6 molecules can also indirectly affect crystallization. For example, previous research reported that InsPs can inhibit osteoclastogenesis and mineralization of osteoblasts . AnotherThe major limitations of this study are that we did not determine the specific InsPs isomers that formed during the thermal hydrolysis of InsP6, and we did not test the effect of these different isomers. This is simply because these isomers are not readily available or are extremely expensive.Although the lower InsPs appear to be responsible for the inhibition of the crystallization of calcium salts in vivo, our in vitro studies showed that the higher InsPs (InsP6 and InsP5) were the most effective in inhibiting CaOx crystallization and that InsP5 and InsP4 were the most effective in inhibiting BRU crystallization. For the specific in vitro experimental conditions of the present study, the InsPs had very weak effects on HAP crystallization. HAP deposits seem to form by a mechanism totally different to the formation of well-defined crystals, depending on the conditions of each case. The in vitro conditions that we examined do not correspond exactly to the in vivo conditions in which HAP deposits are usually formed.


The craniocervical junction (CCJ) is a complex region. Rarely, arteriovenous fistulas (AVFs) can occur in the CCJ region. Currently, it is accepted that CCJ AVFs should only refer to AVFs at the C1-C2 levels. It is reasonable to assume that high cervical CCJ AVFs are being referred to when discussing CCJ AVFs. High cervical CCJ AVFs can be divided into the following four types: dural AVF, radicular AVF, epidural AVF and perimedullary AVF. Until now, it was difficult to understand high cervical CCJ AVFs and provide a proper treatment for them. Therefore, an updated review of high cervical CCJ AVFs is necessary. In this review, the following issues are discussed: the definition of high cervical CCJ AVFs, vessel anatomy of the CCJ region, angioarchitecture of high cervical CCJ AVFs, treatment options, prognoses and complications. Based on the review and our experience, we found that the four types of high cervical CCJ AVFs share similar clinical and imaging characteristics. Patients may present with intracranial hemorrhage or congestive myelopathy. Treatment, including open surgery and endovascular treatment (EVT), can be used for symptomatic AVFs. Most high cervical CCJ AVFs can be effectively treated with open surgery. EVT remains challenging due to a high rate of incomplete obliteration and complications, and it can only be performed in superselective AVFs with simple angioarchitecture. Appropriate treatment can lead to a good prognosis. ArterioEVT can only be used in certain CCJ AVFs because the feeding arteries of most CCJ AVFs are too small and tortuous, and EVT will result in incomplete obliteration, high recurrence and a risk of complications . Current2.The CCJ region is located between the skull base and the upper cervical spine and is formed by the occipital bone and the first two cervical vertebrae (C1 or atlas and C2 or axis), which span both the lower brainstem and upper cervical cord  2, 11, , 11, 12.Broadly speaking, CCJ AVFs should contain foramen magnum (FM) DAVFs . FM DAVFCCJ AVFs at the C1-C2 levels are different from FM DAVFs, although they include the same elements of FM DAVFs and share a similar drainage pathway . Angioar3.3.1.The arteries involved in the high cervical CCJ region mainly originate from the suboccipital V3 segment of the VA, including the radicular  artery, anterior and posterior spinal arteries (ASA and PSA), posterior inferior cerebellar artery (PICA), anterior and posterior meningeal arteries, and some muscular branches  38–40)..38–40). The high cervical cord has little radicular inflow, and the radicular arteries are often thin or invisible on digital subtracted angiography (DSA)  43, 44), 4443, 43.2.Venous drainage in the high cervical CCJ region is abundant, complex and varied, including the extradural and intradural drainage systems , 49, 50.4.4.1.High cervical CCJ AVFs can be located on the inner or outer surface of the dura mater, on the spinal nerves, or on the spinal cord surface and can be divided into the following four types: DAVFs, radicular AVFs (RAVFs), epidural AVFs (EAVFs), and perimedullary AVFs (PAVFs) , 5, in wHigh cervical CCJ AVFs may be single or multiple AVFs, and less than 10% of AVFs are multiple AVFs , 7, 55. High cervical CCJ AVFs present with multiple feeding arteries connecting to the same draining vein . They arMost CCJ AVFs had intradural venous drainage, which was ventral to the medulla oblongata or spinal cord . NormallThe drainage pattern of CCJ AVFs determines their clinical manifestation. CCJ AVFs with downward drainage mainly result in congestive myelopathy , whereasOf these high cervical CCJ AVFs, DAVF tends to present with venous congestion; Intradural RAVF and PAVF tend to accompany aneurysm in fistula and varix on draining vein and may present with hemorrhage , 44, 67.4.2.4.2.1.DAVFs are the most common AVFs in the high cervical CCJ region, accounting for 2.23% of brain and spinal DAVFs , 7, 33. ASA and PSA can be involved in CCJ DAVFs. According to the feeding arteries and their relationships with the ASA and PSA, Choi et al.  classifi4.2.2.RAVFs are defined as the direct communication between a radicular  artery and a radicular vein on spinal C1-C2 nerve roots  72, 73), 7372, 74.2.3.High cervical CCJ EAVFs are not uncommon . They arHigh cervical CCJ EAVFs without the spinal pial artery as the feeder or intradural drainage are generally benign . Compare4.2.4.High cervical CCJ PAVFs are characterized by direct arteriovenous communication without an intervening nidus on the spinal cord surface that consists of single or multiple holes and drains into a single vein  33, 77), 7733, 75.High cervical CCJ AVFs can often present as acute hemorrhage, myelopathy, brainstem dysfunction, radiculopathy, etc. , 78–81. 5.1.Although EVT reportedly has the benefit of being a minimally invasive procedure, it is not the best choice for high cervical CCJ AVFs , 80, 82.In superselective cases of other CCJ AVFs, EVT can be attempted. For instance, in the report by Alshekhlee et al. , a CCJ P5.2.Compared to EVT, open surgery has a higher rate of obliteration and is still considered first-line management for high cervical CCJ AVFs , 13 4, , 85, 86.During surgery for CCJ AVFs, it is crucial to distinguish the fistula, feeding arteries, draining vein, normal spinal artery, and normal spinal vein . Because6.6.1.Good outcomes are defined by a modified Rankin scale (mRS) score of 0–2. In general, open surgery for high cervical CCJ AVFs can achieve good outcomes in more than 90% of cases  3, 4, 3, 53, 90.Except for successful obliteration of CCJ AVFs by either open surgery or EVT, many factors can affect the prognosis of high cervical CCJ AVFs. In general, patients who present with hemorrhage may have better outcomes than those who present with venous congestion. Of these high cervical CCJ AVFs, DAVFs tend to present with venous congestion, and intradural RAVFs and PAVFs tend to present with hemorrhage; therefore, CCJ intradural and extradural AVF cases tend to have better outcomes than DAVF cases. In addition, younger age is predictive of a favorable outcome , 91. Cer6.2.For high cervical CCJ AVFs, complications were always accompanied by open surgery or EVT, including ischemic and hemorrhagic complications, hydrocephalus, and cerebrospinal fluid leakage , 37, 92.Compared with open surgery, EVT has a higher rate of complications . In the Given the risk of serious complications associated with the treatment of CCJ AVF, Inoue et al.  suggeste7.Rarely, AVFs occur in the CCJ region. Currently, it is accepted that CCJ AVFs should only be used to refer to AVFs at the C1-C2 levels. It is reasonable that the term ‘high cervical CCJ AVFs’ is more appropriate to use when discussing CCJ AVFs. High cervical CCJ AVFs can be divided into DAVFs, RAVFs, EAVFs and PAVFs. These four types of CCJ AVFs share similar clinical and imaging characteristics. When they become symptomatic, treatment is necessary. Most high cervical CCJ AVFs can be effectively treated with open surgery. EVT should only be used in certain AVFs with simple angioarchitecture. After appropriate treatment, a good prognosis can be obtained.JY contributed to the conception and design of the review. HS collection of data. JY and HS contributed to drafting the text and preparing the figures. All authors contributed to the article and approved the submitted version.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.