Graphene and graphene oxide have attracted tremendous interest over the past decade due to their unique and excellent electronic, optical, mechanical, and chemical properties. This review focuses on the functional modification of graphene and graphene oxide. First, the basic structure, preparation methods and properties of graphene and graphene oxide are briefly described. Subsequently, the methods for the reduction of graphene oxide are introduced. Next, the functionalization of graphene and graphene oxide is mainly divided into covalent binding modification, non-covalent binding modification and elemental doping. Then, the properties and application prospects of the modified products are summarized. Finally, the current challenges and future research directions are presented in terms of surface functional modification for graphene and graphene oxide. Graphene and graphene oxide have attracted tremendous interest over the past decade due to their unique and excellent electronic, optical, mechanical, and chemical properties. However, four years later, Krätschmer3 confirmed the cage structure of C60-fullerene. In 1991, Nippon Electric Company (NEC) Ltd. first reported carbon nanotubes and expanded the carbon material family.4 In 2004, Novoselov et al.5 successfully separated graphene from the monolithic state using a micro-computer peeling method, which challenged the scientific understanding of two-dimensional crystals. The structure of graphene is shown in 2 hybrid carbon atoms. It is a two-dimensional carbonaceous material with a hexagonal honeycomb crystal structure. To now, graphene is the thinnest and strongest nanomaterial, with a sheet thickness of 0.34 nm.6 Each carbon atom in graphene is bonded to three adjacent carbon atoms through a σ bond. The remaining p electrons most likely form a π bond with the surrounding atoms due to their failure to form a bond, and the bonding direction is perpendicular to the graphene plane. The structure of graphene is very stable, and its C–C bond length is only 0.142 nm.7 The connection between each carbon atom of graphene is very strong. When an external force is applied to graphene, the atomic surface inside it is deformed and further bent to offset the external force. Thus, there is no rearrangement and misalignment between the carbon atoms, maintaining a consistently stable structure.8 When the electrons of graphene move in the internal orbit, there is no scattering phenomenon due to the interference of foreign atoms or lattice defects.9,10 This unique lattice structure gives graphene various excellent properties. Nowadays, there are numerous methods for the preparation of graphene, but the main preparation methods include mechanical stripping, liquid phase stripping, chemical vapor deposition, epitaxial growth and redox methods.11 Recently, significant research has been conducted on graphene quantum dots,12–14 and carbon doped with other elements, molecules or organic materials.15–18Graphene is a carbon material,20 In recent years, with the further study of GO, scientists have found that it also has excellent properties with rich active oxygen-containing functional groups.21 These oxygen-containing groups or reduced doping elements can be used as catalytic active centers for covalent/non-covalent modification design according to the requirements of specific application fields. In addition, the presence of oxygen-containing groups also broadens the interlayer gap of graphene oxide. It can be functionalized by small molecules or polymer intercalations. At present, great progress has been achieved in the functionalization of graphene oxide. It has been applied in the fields of desalination,22 drug delivery,23 oil–water separation,24 immobilization catalysis,25 solar cells,26 energy storage,27 healthcare,28,29etc.Compared with graphene (G), graphene oxide (GO) has the advantages of low production cost, large-scale production, and easy processing. It is often used as a precursor for the preparation of reduced graphene oxide (RGO).However, the single component graphene material has certain limitations, such as weak electrochemical activity, easy agglomeration and difficult processing, which greatly limit the application of graphene. Therefore, functional modification of graphene and graphene oxide is crucial to expanding their application. The functionalization of graphene and graphene oxide is based on the further modification of their intrinsic structure. Herein, we introduce the functional modification methods based on the intrinsic structural chemical bonds and functional groups of graphene and graphene oxide. Firstly, we introduce the basic structure and properties of graphene and graphene oxide, and then their functionalization based on their surface structure features, which is divided into three types, functional modification of covalent bond bonding, functional modification of non-covalent bond action and element doping modification. Subsequently, we categorize and systematically summarize the reaction process and reaction conditions of typical reaction types and their research methods. Finally, the future of surface functionalization of graphene and graphene oxide is presented.2.x electron and 2py electron), forming plane sp2 hybrid orbitals. The remaining orbital electron forms a large π bond, and this electron can move freely in the plane. Graphene and graphene oxide show excellent electrical, mechanical, and thermal properties due to their unique structural and morphological features.30–36 They exhibit the Hall effect, tunneling effect, bipolar electric field effect and high thermal conductivity. The electrons are not easily scattered when they are transported in G, and the maximum electron mobility at room temperature can reach 2 × 105 cm2 (V s)−1,24 and thus the ideal conductivity of G is over 1 × 106 S cm−1.24 The Young's modulus of G can reach up to 1100 GPa,25 its transmittance is about 97.9% for visible light,25 and its specific surface area can be as high as 2630 m2 g−1.26 As a two-dimensional carbon material, graphene oxide has a single atomic layer, and the size of its sheets is polydisperse. Compared with graphene, there are many oxygen-containing functional groups on the graphene oxide sheet layer, thus the structure of graphene oxide is more complicated, and its properties depend on its structure. Lerf and Klinowski37,38 put up the structural model of graphene oxide, called the L–K model. It indicates that the hydroxyl and epoxy groups are randomly distributed on the graphene oxide single layer, while the carboxyl and carbonyl groups are introduced at the edge of the single layer. Graphene oxide has an unoxidized benzene ring region and an oxidized aliphatic six-membered ring region, and the relative size of these two regions depends on the degree of oxidation and random distribution on graphene oxide. However, the L–K structural model is based on certain conditions, ignoring the influence of raw graphene, oxidant and oxidation methods. Erickson et al.39 studied graphene oxide nano-plates via scanning electron microscopy (SEM), and found that graphene oxide not only has an oxidized region with high disorder and unoxidized graphene region, but also has hole defects due to overoxidation and lamellar peeling. Accordingly, in recent years, other models have been proposed, such as the dynamic structure model (DSM)40,41 and binary structure model.42 Extensive research has been conducted on the preparation and properties of G and GO by many scientists, and it has been found that the obvious difference between graphene and graphene oxide is the addition of oxygen atoms bound with some carbons, as shown in 2) and aliphatic (sp3) domains, which lead to an increase in the type of interactions that can occur on its surface. Graphene oxide can be reduced to graphene by a reducing agent. However, the produced graphene is not suitable for electronic applications and mechanical reinforcement of polymers due to the structural defects created during the synthesis of graphene oxide. Nevertheless, this is the preferable route for the large-scale modification of the surface properties of graphene materials by functionalization.Graphene has a planar hexagonal lattice structure. There are 4 valence electrons per carbon atom, including 3 electrons  had strong oxidizing properties in concentrated sulfuric acid, and there was a high risk of explosion when Mn2O7 contacted with organic matter at 55 °C.55 Subsequently, Marcano50 used the less corrosive phosphoric acid to produce graphene oxide in situ. This method does not involve the release of intense exothermic or toxic gases, and the prepared graphene oxide has a higher degree of oxidation and structural regularity, but requires a large amount of KMnO4 and concentrated sulfuric acid, thus increasing the cost of processing raw materials and the treatment of waste liquids. The preparation of graphene oxide using the K2FeO4/H2SO4 oxidation system49 at room temperature for 1 h is a safe and efficient method. Using benzoyl peroxide52 as an oxidant at 110 °C, graphene oxide can also be prepared by reaction for 10 min. Although this method is highly efficient, benzoyl peroxide itself is extremely unstable and there may be a risk of explosion when the reaction temperature is high. Besides the effect of oxidants on graphene oxide, the graphite raw materials and the reaction methods also affect the properties of graphene oxide, where relatively few hydroxyl and carboxyl groups are obtained on graphene oxide prepared from highly crystalline graphite powder.56 During the low-temperature and mid-temperature stages of graphene oxide preparation, a modified Hummers' method with the help of auxiliary ultrasound equipment was used, and the layer spacing of graphene oxide became larger.57 However, there were many holes in the graphene oxide due to excessive oxidation. When the reaction temperature was over 50 °C, the graphene oxide became unstable. If this reaction occurred at a relatively low temperature, the hole defects of graphene oxide would diminish significantly.58 Currently, the feasibility of the industrial preparation of graphene with graphene oxide as the precursor is high, but the existing graphene oxide preparation methods generally have technical problems such as complicated purification steps and many defects. Therefore, compared with mechanically exfoliated high-quality graphene, the prepared reduced graphene oxide is inferior in structure and properties.The different methods have advantages and disadvantages. In the early stages, nitric acid and KClO59 and preserves their inherent properties. The functionalization of graphene and graphene oxide results in new functions, and subsequently can possess excellent mechanical properties, electrical properties, thermal properties, etc. Currently, the methods for the functionalization of graphene and graphene oxide mainly include covalent bond functionalization, non-covalent bond functionalization, and other atomic doping functionalization.60The proper functionalization of graphene and graphene oxide prevents agglomeration during the reduction of graphene and graphene oxide,4.4.1et al.61 prepared graphene from graphene oxide using the sodium borohydride reducing agent and then sulfonation with the aryl diazonium salt of sulfanilic acid. The light sulfonated graphene was easily dispersed in water at a suitable concentration (2 mg mL−1) in the pH range of 3–10. By observing the AFM images shown in Si et al.62 reported a simple method for the fabrication of multifunctional fibers with mechanically strong RGO cores and highly conductive chemical vapor deposition (CVD) graphene shells (rGO@Gr fibers), which showed outstanding electrical conductivity as high as 137 S cm−1 and a failure strain value of 21%. These values are believed to be the highest values among polymer-free graphene fibers. We also demonstrated the use of the rGO@Gr fibers in high power density supercapacitors with enhanced mechanical stability and durability, which enables their practical applications in various smart wearable devices in the future. The main method involved the preparation of the CVD graphene film, GO fiber fabrication, fabrication of graphene–GO fiber (a reduction solution (HI : AcOH = 2 : 5 v/v)) and testing the thermal conductivity.Choi 4.2The reduction of GO with an alcohol is a relatively mild reduction method because this treatment method does not cause severe damage to the edge morphology of GO, and highly conductive RGO can be prepared.et al.63 reduced GO with ethanol vapor at high temperature to obtain highly conductive RGO. Experiments showed that the impedance of GO was 188–418 kΩ μm−2, and after ethanol vapor reduction at 900 °C, the impedance was reduced by 43 kΩ μm−2. This result was similar to the result from reducing GO at 1100 °C under vacuum (40–100 kΩ μm−2). Besides, they also studied the reduction of GO with H2 at high temperature. The results indicated that ethanol vapor was more effective at reducing GO at the same temperature.Su 4.3et al.64 studied the peeling of GO for 20 h into a GNS sheet by hydroquinone upon refluxing GO with hydroquinone. The results showed that the product RGO had a low oxygen content and maintained an ordered crystal structure, but agglomeration occurred within only a few hours in water. It was also found that the thermal stability of the product was worse than that of graphite powder.In addition to alcohol reducing agents, some phenolic reducing agents are also used to reduce GO. Wang 4.4via the rapid heating of dry GO under inert gas and high temperature. The properties of thermal reduction products are directly related to the heating rate, reduction temperature and time. Too fast heating rate and too long reduction time will greatly expand the volume of the product, and the specific surface area is much lower than the theoretical value of graphene. Hyunwoo Kim65 reviewed that in top-down processes, graphene or modified graphene sheets are produced by separation/exfoliation of graphite or graphite derivatives. In general, these methods are suitable for large-scale production for polymer composite applications. For the process of TRG, heating GO in an inert environment for 30 s at 1000 °C causes the reduction and flaking of GO, thereby producing TRG sheets. Delamination occurs when the pressure generated by the gas (CO2) due to the decomposition of the epoxy and hydroxyl groups of GO exceeds the van der Waals force holding the graphene oxide sheets together.Thermally reduced graphene oxide (TRG) can be produced 66 also reported a polymer/GO material, where its preparation involved aqueous mini-emulsion copolymerization of St and nBA in the presence of nano-dimensional GO  and the conventional surfactant SDS. The obtained latex comprising polymer particles armoured with GO sheets were used directly for film formation at ambient temperature.Fadil4.52 reduction, ultraviolet radiation reduction method, and electrochemical reduction method. Some researchers prepared RGO using sulfur-containing compound reducing agents, and found that sulfur-containing compounds have good reducing properties. The oxygen content in the reduced product is relatively low and stably dispersed in solution. When SO2 gas was used as a reducing agent for reducing GO, the oxygen content of the product RGO obtained by this method was similar to that using hydrazine as the reduction agent, and the sulfur-containing compound could promote the dispersion of the product in aqueous solution.67 It was also found that the product RGO was a single layer having a thickness of only 0.87 nm, but the surface had significant wrinkles. When the number of layers was below 10 layers, a large area of wrinkles appears in GO,68 as shown in There as other reduction methods such as sulfur-containing compound reducing agent reduction method, SOet al.69 conducted an in-depth study on the ultraviolet (UV) irradiation reduction method, and a schematic of the TiO2–graphene composite and its response under UV-excitation is shown in Ultraviolet radiation reduction and electrochemical methods are also environmentally friendly methods for the preparation of GNS, and they have a great potential in the large-scale preparation of GNS. Williams They obtained stable RGO by reducing the GO suspension by UV irradiation. Compared with the use of a photocatalyst or chemical reducing agent, this method is not only simple and easy, but also inexpensive, and it provides a new possible way for the mass preparation of GNS.5.59 and preserves their inherent properties. The functional modification of graphene and graphene oxide not only maintains their excellent characteristics, but also introduces new functional groups to give them new characteristics. Also, different functional groups give different characteristics. Presently, the methods for the functionalization of graphene and graphene oxide mainly include covalently functionalization, non-covalent functionalization, and elemental doping.70The proper functionalization of graphene and graphene oxide prevents their agglomeration during the reduction process,5.171 The covalent bond functional modification of graphene and graphene oxide is illustrated below according to the functional group. Covalent bond modification increases the processability and brings new functions. There are multiple strategies to covalently functionalize GO. For example, Man et al.72 using polystyrene particles “armoured” with nanosized graphene oxide (GO) sheets, which were prepared by aqueous mini-emulsion polymerization of styrene, exploiting the amphiphilic properties of GO in the absence of conventional surfactants. The nanoscale GO sheets were prepared from graphite nanofibers with a diameter of approximately 100 nm based on a novel procedure, thus effectively ensuring the absence of larger sheets.Covalent bond functionalization of graphene involves combining graphene with newly introduced groups in the form of covalent bonds to improve and enhance its performance. The oxygen-containing groups on the surface of graphene oxide makes covalent bond functionalization easier than that on graphene. The surface of graphene oxide contains a large amount of hydroxyl groups, carboxyl groups, and epoxy groups. These groups can be used for common chemical reactions such as isocyanation, carboxylic acylation, epoxy ring opening, diazotization, and addition.5.1.1 PBM data was replaced with SVG by xgml2pxml:<glyph-data id="z.dbd" format="PBM" resolution="300" x-size="8" y-size="10" xml:space="preserve">00000000000000000000000000000000111111110000000011111111000000000000000000000000</glyph-data><svg xmlns="http://www.w3.org/2000/svg" version="1.0" width="13.200000pt" height="16.000000pt" viewBox="0 0 13.200000 16.000000" preserveAspectRatio="xMidYMid meet"><metadata>Created by potrace 1.16, written by Peter Selinger 2001-2019</metadata><g transform="translate scale" fill="currentColor" stroke="none"><path d="M0 440 l0 -40 320 0 320 0 0 40 0 40 -320 0 -320 0 0 -40z M0 280 l0 -40 320 0 320 0 0 40 0 40 -320 0 -320 0 0 -40z"/></g></svg>C bond in the aromatic ring of graphene or graphene oxide. The graphene oxide diazotization reaction and Diels–Alder reaction have been reported.73–75The functional modification of the carbon skeleton is mainly carried out using the Cet al.76 used solution-phase graphene as a raw material dispersed in 2% sodium cholate (as surfactants) aqueous solution, and stirred it with 4-propargyloxydiazobenzenetetrafluoroborate at 45 °C for about 8 h, to obtain 4-propargyloxyphenyl graphene  scale" fill="currentColor" stroke="none"><path d="M80 600 l0 -40 600 0 600 0 0 40 0 40 -600 0 -600 0 0 -40z M80 440 l0 -40 600 0 600 0 0 40 0 40 -600 0 -600 0 0 -40z M80 280 l0 -40 600 0 600 0 0 40 0 40 -600 0 -600 0 0 -40z"/></g></svg>CH). Then, a click chemistry reaction with the azido polyethylene glycol carboxylic acid . Finally, alkyne-functionalized polystyrene (HCC–PS) was used to graft polystyrene onto the surface of the oxide by esterification to obtain graphene-based polystyrene. The modified graphene oxide had good solubility in polar solvents such as tetrahydrofuran, dimethylformamide and chloroform, and the distance between GO layers could be controlled by the length of PS. This method can be extended to the functionalization of other graphite polymer composites. The preparation route and reaction conditions for the target hydroxy-based functionalization of graphene oxide is shown in Graphene oxide contains a large amount of reactive hydroxyl groups on its sheet layer, where hydroxyl-based functional modification generally involves hydroxy-reaction of amide or isocyanate and graphene oxide to produce esters, and then further functional modification using different groups.et al.80 demonstrated novel reversible addition–fragmentation chain transfer agent (RAFT-CTA)-modified reduced graphene oxide nanosheets (CTA-rGONSs) by crosslinking rGONSs with a RAFT-CTA via esterification reaction. These nano CTA-rGONSs were used to polymerize a hydrophobic amino acid-based methacrylamide  monomer with different monomer/initiator ratios. The synthetic procedure is shown in detail in Namvari 5.1.381,82 The carboxyl functionalization step is generally the activation of the reaction, and then the group containing an amino group and a hydroxyl group is dehydrated to form an ester or amide bond. The reagents commonly used for carboxyl activation include thionyl chloride (SOCl2),83 2-(7-aza-1H-benzotriazole-1-yl)-1,1,3,3-tetramethyluronium hexafluorophosphate,84N,N-dicyclohexylcarbodiimide (DCC),85 and 1-ethyl-3-(3-dimethylaminopropyl)-carbodiimide (EDC).86 Ouyang et al.87 demonstrated the chemical functionalization of semiconducting graphene nanoribbons (GNRs) with Stone–Wales (SW) defects by carboxyl (COOH) groups. It was found that the geometrical structures and electronic properties of GNRs changed significantly, and the electrical conductivity of the system could be considerably enhanced by mono-adsorption and double adsorption of COOH, which sensitively depends on the axial concentration of SW defect COOH pairs (SWDCPs). With an increase of the axial concentration of SWDCPs, the system transformed from semiconducting behavior to p-type metallic behavior. This makes GNRs a possible candidate for application in chemical sensors and nanoelectronic devices. The preparation route and reaction conditions for the target hydroxy-based functionalization of graphene oxide are shown in There are a large number of carboxyl groups at the edge of graphene oxide, and the carboxyl group is a highly reactive group, and thus has been greatly studied for the functionalization of graphene oxide.et al.88 removed the oxygen-containing groups from GO through a reductive treatment, and consequently reintroduced carboxyl groups onto the graphene surface. Carboxyl groups were inserted based on a free-radical-addition reaction, which occurred not only on the carbon atoms located at the edge plane, but also at the much more abundant basal plane of the graphene sheets. Specifically, chemically reduced graphene oxide (CRGO) was first functionalized with isobutyronitrile groups, which were generated from the thermal decomposition of azobisisobutyronitrile (AIBN) to give CRGO–CN. Subsequent reflux of CRGO–CN in a mixture of methanol and sodium hydroxide aqueous solution resulted in a hydrolysis reaction to provide CRGO enriched with carboxyl groups (CRGO–COOH), as shown in Bonanni 5.2The non-covalent bond functionalization of graphene or graphene oxide results in the formation of a composite material having a specific function by interaction between hydrogen bonds and electrostatic forces between graphene and functional molecules, the greatest advantage of which is maintaining the bulk structure and excellent properties of graphene or graphene oxide, and also improving the dispersibility and stability of graphene or graphene oxide. The methods for the functional modification of surface non-covalent bonds mainly include π–π bond interaction, hydrogen bonding, ionic bonding, and electrostatic interaction modification. The non-covalent bond functionalization process is simple with mild conditions, while maintaining the structure and properties of graphene. However, the disadvantage of this method is that other components (such as surfactants) are introduced.5.2.1et al.89 inspired by interfacial interactions of protein matrix and the crystal platelets in nacre, produced a super tough artificial nacre through the synergistic interface interactions of p–p interaction and hydrogen bonding between graphene oxide (GO) nanosheets and sulfonated styrene–ethylene/butylene–styrene copolymer synthesized with multifunctional benzene. The resultant GO-based artificial nacre showed super-high toughness of 15.3 ± 2.5 MJ m−3, superior to that of natural nacre and other GO-based nanocomposites. The ultra-tough property of the novel nacre was attributed to the synergistic effect of the p–p stacking interactions and hydrogen bonding. Thus, this bioinspired synergistic toughening strategy opens a new avenue for constructing high performance GO-based nanocomposites in the near future.Song et al.90 used a tetradecene derivative with a dendritic polyether branch as a modifier and the synergistic effect of an aromatic cyclic fluorene skeleton interacting with graphite and a polyether chain to induce high hydrophilicity, stripping graphite and stabilizing the graphene layer, as shown in Lee 5.2.2et al.91 researched the fabrication of reduced graphene oxide aerogel membranes (rGOAMs) via the reduction-induced self-assembly of rGO through hydrogen bond mediation. Using polyethylene glycol (PEG) as the mediator, the PEG–rGO hydrogen bonding interactions partly replaced the interlayer p–p and hydrophobic interactions during reduction, decreasing the rGO laminate size in 2D stacking and alleviating the structural shrinkage of the rGOA networks. The tight correlation between membrane pore size and porosity was broken, leading to rGOAMs with tunable pore sizes (0.62 to 0.33 mm) and high porosity (95%). The resultant rGOAMs could effectively reject oil-in-water emulsions with different sizes and exhibited ultrahigh water flux (up to 4890 L m−2 h−1) under 0.10 bar as well as a persistent anti-oil-fouling performance for up to 6 cycles.He et al.92 realized the surface functionalization of graphene by hydrogen bonding between graphene and DNA, which improved the hydrophilicity of graphene and stabilized it in water. On the other hand, the loading of organic molecules occurred on the surface of graphene. The functional modification of the graphene surface by hydrogen bonding does not introduce impurities, which is safe and reliable, and has important potential application prospects in biomedical field. A schematic illustration of aqueous dispersions of (a) GO, (b) GO–PDI and (c) GO–PyS through π–π interaction is shown in Patil via a simple noncovalent method, and the loading and release behaviors of DXR on GO were investigated by Patil et al.93 The efficient loading of DXR on GO was as high as 2.35 mg mg−1 at the initial DXR concentration of 0.47 mg mL−1. The loading and release of DXR on GO showed strong pH dependence, which may be due to the hydrogen-bonding interaction between GO and DXR. The fluorescent spectrum and electrochemical results indicate that strong π–π stacking interaction exists between them, as shown in A novel graphene oxide–doxorubicin hydrochloride nanohybrid (GO–DXR) was prepared 5.2.3et al.94 reported that a stable dispersion of reduced graphene was achieved in various organic solvents via noncovalent ionic interaction functionalization with amine-terminated polymers. An aqueous dispersion of reduced graphene was prepared via the chemical reduction of graphene oxide in aqueous media and vacuum filtered to generate reduced graphene sheets. Thorough FTIR and Raman spectroscopy investigation verified that the protonated amine terminal group of polystyrene underwent noncovalent functionalization with the carboxylate groups on the graphene surface, providing high dispersibility in various organic media, as shown in Choi et al.95 improved the mechanical and barrier properties of a starch film with reduced graphene oxide (RGO) modified by sodium dodecyl benzene sulfonate (SDBS). The hydrophilia of the modified RGO (r-RGO) was improved, which resulted in its good dispersion in the oxidized starch (OS) matrix. The tensile strength of the r-RGO-4/OS film increased to 58.5 MPa, which was more than three times that of the OS film (17.2 MPa). Besides, both the water vapor and oxygen barrier properties of the r-RGO/OS film improved greatly compared with that of the OS and GO/OS films. Moreover, the r-RGO/OS film could protect against UV light effectively due to its lightproof performance. In conclusion, the r-RGO/OS composite film has great potential applications in the packaging industry.Ge 5.2.4et al.60 used hydrazine as a reducing agent to control the reduction, while removing the functional groups such as hydroxyl groups and epoxy bonds of graphene oxide and retaining the carboxyl anion, which is well dispersed by charge repulsion. The chemical conversion of graphene can be conducted in water. Graphene oxide is soluble in water because its surface negative charge repels each other and forms a stable colloidal solution, as shown in Electrostatic repulsion between the same type of charge is another strategy to improve the dispersion of graphene. Bhunia 96 prepared novel environmentally friendly Gemini surfactants, i.e. each with two hydrophilic and two hydrophobic groups. The reaction was conducted in hydrogen peroxide for the epoxidation of the carbon–carbon double bond. Methylene was used as the spacer group, and the nonionic hydrophilic head group was then introduced by ring-opening reaction. The targeted surfactants were synthesized via the reaction of chlorine sulfonic acid and hydroxy esterification. The results were compared with that for mixtures of the standard surfactants sodium decylsulfate and octaoxyethyleneglycol mono n-decyl ether under equivalent conditions. The surfactants were shown to exhibit improved performance over the mixed system both in terms of micellization and surface tension lowering.Ge X. and Li H.5.397–100 Element doping adjusts the energy band structure of graphene, but the doping process is difficult to control quantitatively.Element doping modification usually adopts annealing heat treatment, ion bombardment, arc discharge and other means to incorporate different elements into graphene, thereby resulting in the substitution of defects and vacancy defects in graphene, and maintaining the intrinsic two-dimensional structure of graphene. Simultaneously, its surface properties change to give new performances.et al.100 used thermal annealing to treat graphene and ammonium nitrate to prepare 6.54 at% nitrogen-doped graphene, which catalyzed the oxidative degradation of phenol by 5.4 times that of undoped graphene. The synergistic effect of B-, P- or N-doped graphene was studied. The detailed growth process of N-doped graphene by thermal annealing treatment is shown in Duan et al.101 reported a CVD technique to produce N-doped graphene, which was the first experimental example of substitutionally doped graphene, which was hard to produce using other methods. The CVD method is a nondestructive route to produce graphene and realizes substitutional doping since the doping is accompanied by the recombination of the carbon atoms in graphene during the CVD process. By using SEM, TEM, Raman, XPS and EDS, they demonstrated the existence of N-doped graphene. The CVD method can not only be used to produce N-doped graphene, but also has potential to produce the graphene doped with other elements. Moreover, they measured the electrical properties of the N-doped graphene. This research provides a new type of graphene experimentally, which is required for the further application of graphene. The synthesis of N-doped graphene by electrostatic repulsion is shown in Wei 6.The functional modification of graphene is of great significance for the application of graphene composites. According to previously mentioned literature, the surface functional modification of graphene and graphene oxide is used to obtain related products and the interactions and reactions of the modification types have been studied, briefly explaining the functional characteristics and application fields of graphene surface-functionalized composites, as summarized in 7.102 medicine,103 energy,104 water splitting, biosensing, bioimaging, environmental studies, catalysis, photocatalysis, and biomedical technologies.105,106 However, a coin has two sides. Because of the unique structure of graphene, it is difficult to disperse in water and organic solvents, which is known as the agglomeration phenomenon of graphene. This problem limits the application of graphene. Therefore, it is important to functionalize graphene and graphene oxide to expand their various applications.Graphene has many unique and prominent physical and chemical properties  due to its special structure, and thus has wide application potential in many fields, such as heterojunction solar cells,Currently, the study of these functional methods is still in the experimental stage, and researchers are mainly focused on the development of new product varieties and their characterization and application. However, there are several problems as follows. (1) Functionalized target groups difficult to control, (2) the synthetic procedures are complicated, and (3) separation and purification are very difficult.Thus, considering the above-mentioned superior performance, we should pay attention to the relationship between the structure and properties of graphene and graphene oxide in future work. For example, according to the quantitative structure and property relationship principle, functional groups are introduced into molecules based on functionalized target group design. Functionalized target group design is carried out by molecular simulation technology, and the structure property model is set up to guide the synthesis of new products. However, the synthetic process of different types of graphene and graphene oxide is complex, which involves the introduction of many other special groups, thus it is difficult to control the reaction and obtain the target products. Also, some reaction conditions are extremely harsh. Nevertheless, considering the optimization of reaction procedures and environmentally friendly synthetic routes, graphene and graphene oxide will achieve industrialization. Of course, other new functional modification methods should be developed, such as non-alkali carbonylation and carbon–carbon multi-bond addition reactions, which are rarely reported in functional applications.8.Graphene-based composites are involved in many fields with the development of science, such as graphene-based drug carriers to improve the stability, toxicity, metabolic kinetics of drugs, and many aspects are still unclear, pending further research and exploration by scientists. Accordingly, graphene and graphene oxide will be researched completely and use widely in the future.There are no conflicts to declare.


Central nervous system (CNS) metastases and acquired resistance complicate the treatment of anaplastic lymphoma kinase (ALK) rearrangement-positive (ALK-p) advanced non-small cell lung cancer (NSCLC). Thus, this review aimed to provide a comprehensive overview of brain metastasis, acquired resistance, and prospects for overcoming these challenges. A network meta-analysis of relevant phase III randomized controlled trials was performed to compare the efficacies of multiple ALK inhibitors by drug and generation in overall patients with ALK-p untreated advanced NSCLC and a subgroup of patients with CNS metastases. The primary endpoint was progression-free survival (PFS). Generation-specific comparison results showed that third-generation ALK inhibitors were significantly more effective than second-generation ALK inhibitors in prolonging the PFS of the subgroup of patients with CNS metastases. Drug-specific comparison results demonstrated that lorlatinib was the most effective in prolonging PFS, followed by brigatinib, alectinib, ensartinib, ceritinib, crizotinib, and chemotherapy. While lorlatinib was superior to brigatinib for PFS in the overall patient population, no significant difference between the two was found in the subgroup of patients with CNS metastases. These results can serve as a foundation for basic, clinical, and translational research and guide clinical oncologists in developing individualized treatment strategies for patients with ALK-p, ALK inhibitor-naive advanced NSCLC. The tremendous advances in tumor molecular research over the past two decades have contributed enormously to our understanding of the etiology of non-small cell lung cancer (NSCLC), which constitutes 84% of all primary lung cancers ,2,3,4,5.As shown in Lung cancer has a high probability of metastasis to the CNS, and patients with advanced NSCLC involving driver gene alterations, such as EGFR mutations and ALK rearrangements, frequently have brain metastases (BM). The incidence of BM in such patients is 37–64% ,24,25. HNevertheless, therapeutic strategies targeting ALK-p NSCLC with CNS metastases remain to be developed, and current strategies need to be improved ,12,14.Even patients who respond well to initial therapy with ALK inhibitors may experience tumor recurrence because of acquired resistance to these drugs via multiple mechanisms, including secondary ALK resistance mutations and tumor growth promoting molecular pathways. Therefore, the mechanisms underlying acquired resistance to ALK inhibitors must be elucidated to develop novel strategies to treat patients with ALK-p advanced NSCLC, especially those with CNS metastases ,12.In this meta-analysis report we first summarize the different ALK inhibitors  used in patients with ALK-p advanced NSCLC. We then discussed the treatment of patients with CNS metastases and the prospects for overcoming acquired resistance mutations in these patients.ALK inhibitors bind to the ATP-binding pocket of the intracellular tyrosine kinase domain, and regulate their downstream signals such as the RAS, PI3K-AKT, JAK/STAT signaling cascades which are involved in tumor progression; attenuation of these cascades produces an antitumor effect ,20,27. ACrizotinib is a first generation ALK inhibitor authorized for the potential therapeutic application in the treatment of ALK-p NSCLC. Two phase III trials (PROFILE1014  and PROFCeritinib is a second-generation ALK inhibitor developed to improve the low activity of first-generation ALK-TKIs against CNS diseases and overcome resistance. A phase III study (ASCEND-4) found that the PFS of patients with stage IV ALK-p NSCLC and PS 0–1 is longer with ceritinib monotherapy than with platinum combination therapy  . HoweverThree phase III trials  that inL  that in = 7/11 patients) than that of crizotinib monotherapy . However, the incidence of treatment-related serious adverse events is higher with ensartinib monotherapy (7.7%) than with crizotinib monotherapy (6.1%), with no new safety signals [A phase III eXalt3 trial demonstrated that ensartinib (X-396) monotherapy, as a first-line treatment for patients with ALK-p advanced NSCLC, is better than crizotinib monotherapy in prolonging PFS  . In addi signals .A phase III trial (CROWN ) showed In this section we discuss the treatment options for asymptomatic and symptomatic BM cases and the future direction of drug therapy development.For asymptomatic BM cases, single-drug therapy with TKIs is the preferred and recommended treatment option because the patients’ tumors, including BM, are expected to have high responsiveness to these drugs ,26.However, BM progression can easily cause neurological symptoms and rapidly deteriorate the patient’s condition. For example, BM lesions in the brain stem or close to the pyramidal tract can rapidly deteriorate, even if their size is small. Thus, intracranial radiotherapy is preferred for such patients, even those who are asymptomatic. Close monitoring of BM and the timing of radiotherapy intervention are critical in managing asymptomatic cases ,12,14. For symptomatic BM cases intracranial radiotherapy is the primary treatment option because robust local control of BM and neurological improvement are expected. The drugs selected by treating physicians are also essential in managing symptomatic BM cases. Several phase III clinical trials demonstrated that the intracranial antitumor activity of second- or further-generation ALK-TKIs is higher than that of crizotinib ,43. LorlDrug delivery into the CNS is generally prevented and primarily regulated by the BBB. WBRT can irreversibly disrupt the BBB and improve the delivery of ALK-TKIs ,47. WBRTDrug therapy combined with other agents and fourth-generation ALK-TKIs are future treatment options for ALK-p NSCLC with BM.Glycogen synthase kinase 3 (GSK3) is a serine-threonine kinase that serves an essential role in many cellular processes; thus, it has been considered as a promising target in various malignant diseases including brain tumors . A preclRadiotherapy, drug therapy, and their combinations have markedly progressed and prolonged the survival of patients with BM from ALK-p NSCLC. However, individualized treatment selection is a critical component for managing such patients. Thus, a multidisciplinary team comprising thoracic and radiation oncologists should conduct individualized treatment discussions to prolong the survival of patients with BM from ALK-p NSCLC.Although crizotinib can significantly prolong the response rate and PFS of patients with ALK-p NSCLC, disease progression inevitably occurs after treatment because of the acquired resistance of 1–2 years . The mecSecondary ALK inhibitor resistance mutations were previously identified in 20–30% of tumor samples with crizotinib failure . Among tSecond-generation ALK inhibitors, such as ceritinib, alectinib, and brigatinib, have been developed and approved clinically to overcome crizotinib-resistant mutations. These inhibitors are potent against common crizotinib-resistant mutations, L1196M, and G1269A ,59,60,61Although the G1202R mutation is rarely detected in crizotinib-relapsed clinical samples (2%), it is the most frequent resistance mutation following the administration of second-generation ALK inhibitors, accounting for 40–65% of all acquired resistance mutations ,64,65. SIn 2018 lorlatinib was approved by the U.S. FDA for the treatment of patients with ALK-p NSCLC. Lorlatinib displays activity against all potential ALK-TKI resistance mutations, including L1196M, G1269A, and G1202R . LorlatiMDR1) gene can induce multidrug resistance through the ATP-dependent efflux of chemotherapeutical agents [Activating bypass signaling pathways is important in ALK-independent resistance mechanisms via gene alterations, autocrine signaling with ligand overexpression, and feedback signaling. Such pathways include epidermal growth factor receptor signaling ,72, KIT l agents . P-gp acl agents . In mostl agents . Brain al agents ; otherwil agents . Phenotyl agents ,84,85,86l agents . The hisl agents . With reBased on these prospects, we conducted a comprehensive literature search and network meta-analysis . The results of this meta-analysis provide important information to guide clinical oncologists treating non-small cell lung cancer when considering treatment strategies for patients with ALK-p, ALK inhibitor-naive advanced NSCLC.A systematic literature search identified 2724 studies , second-generation ALK inhibitors , and third-generation ALK inhibitors (lorlatinib) in prolonging PFS were compared between generations in the overall patient population of ALK-p, ALK inhibitor-naive advanced NSCLC. Statistically significant differences were found among all generations compared  . RankingThe efficacies of chemotherapy, first-generation ALK inhibitors (crizotinib), second-generation ALK inhibitors , and third-generation ALK inhibitor (lorlatinib) in prolonging the PFS in a subgroup of patients with CNS metastases were compared. Statistically significant differences were found between third- and second-generation ALK inhibitors, third- and first-generation ALK inhibitors, second- and first-generation ALK inhibitors, third-generation ALK inhibitors and chemotherapy, and second-generation ALK inhibitors and chemotherapy, but not between first-generation ALK inhibitors and chemotherapy . RankingA paired comparison of the efficacies of ensartinib, lorlatinib, brigatinib, alectinib, ceritinib, crizotinib, and chemotherapy in prolonging the PFS of the overall patients is presented in A paired comparison of the efficacies of ensartinib, lorlatinib, brigatinib, alectinib, ceritinib, crizotinib, and chemotherapy in prolonging the PFS in the subgroup of patients with CNS metastases is presented in A paired comparison of the efficacies of ensartinib, lorlatinib, brigatinib, alectinib, ceritinib, crizotinib, and chemotherapy in prolonging the PFS in non-Asian subgroup was presented in A paired comparison of the efficacies of ensartinib, lorlatinib, brigatinib, alectinib, ceritinib, crizotinib, and chemotherapy in prolonging the PFS in the subgroup of patients with CNS metastases is presented in The qualities of the studies that were included were appraised on the basis of the Cochrane-recommended Risk of Bias tool 2 (RoB2) . Nine stOf the nine studies included in this analysis, three , indicating mild between-trial heterogeneity  were compared with that of chemotherapy in the overall patients with ALK-p, ALK inhibitor-naïve advanced NSCLC and in the subgroup of patients with CNS metastases. The comparisons were conducted by ALK inhibitor generation and by drug. Generation-specific comparison results showed that third-generation ALK inhibitors had the best efficacy in prolonging the PFS of the overall patient population and in the subgroup of patients with CNS metastases. Statistically significant differences in efficacy were found between third-generation ALK inhibitors and second- and first-generation ALK inhibitors in the overall patients and in the subgroup of patients with CNS metastases. Significant differences in efficacy in prolonging PFS were also demonstrated between second- and first-generation ALK inhibitors. Drug-specific comparison results showed that lorlatinib had the best efficacy in prolonging PFS in the overall patients, followed by alectinib, brigatinib, ensartinib, ceritinib, crizotinib, and chemotherapy. The differences between lorlatinib and brigatinib, ensartinib, ceritinib, crizotinib, and chemotherapy were statistically significant. Analysis of the subgroup of patients with CNS metastases showed that lorlatinib exerted the most favorable effect, followed by brigatinib, ensartinib, ceritinib, crizotinib, and chemotherapy. The differences between lorlatinib and ensartinib, ceritinib, crizotinib, and chemotherapy were statistically significant. Several previous meta-analyses have compared the efficacies of ALK inhibitors in patients with ALK-p, ALK inhibitor-naive advanced NSCLC ,103,104.Notable findings were also obtained in drug-specific comparisons. For instance, lorlatinib was significantly better than brigatinib in prolonging the PFS of the overall patients, but their difference was not significant in the subgroup of patients with CNS metastases. In addition, evaluation results showed that brigatinib ranked third in the overall participant population but second, above alectinib, in the subgroup of patients with CNS metastases. These results support the theory that brigatinib, along with lorlatinib, is a potential first-line treatment option for ALK-p, ALK inhibitor-naïve advanced NSCLC with CNS metastases.Our results also suggest that lorlatinib has potential as a novel first-line treatment for ALK-p, ALK inhibitor-naive advanced NSCLC. However, lorlatinib should not be recommended for all patients with this disease because its tolerability is reportedly lower than that of alectinib, and its effect on OS was not evaluated. Furthermore, in our analysis of racial differences lorlatinib ranked highest in PFS among non-Asians, whereas ensartinib ranked highest among Asians. Further clinical studies are warranted to develop a detailed treatment strategy for first-line treatment of ALK-p ALK-untreated advanced NSCLC.This NMA has several limitations. First, the study compared the efficacies of six ALK inhibitors in the overall patients and subgroup of patients with CNS metastases. However, OS and safety outcomes were not analyzed because of insufficient data reported for CNS metastases. Further validation is needed to determine whether the results of this comparative analysis of PFS in the subgroup with CNS metastases will be consistent with the results of the comparative analyses of OS and safety outcomes. Second, this analysis included patients who had received systemic anticancer chemotherapy and those who had not. Although the results of sensitivity analysis showed that the inclusion and exclusion of patients who had undergone systemic anti-cancer chemotherapy did not apparently influence the results, we cannot completely rule out the potential impact of this heterogeneity on the final conclusions. Third, mild heterogeneity, although not statistically significant, was demonstrated in the five studies comparing second-generation ALK inhibitors with crizotinib  age 18 years or older; (2) histologically or cytologically confirmed progressive or metastatic ALK-p NSCLC; (3) performance status of 0 to 2 ; (4) at least one measurable lesion assessed according to RECIST version 1.1,25; and (5) no prior exposure to ALK-targeted therapy.In this analysis, patients treated with ensartinib (225 mg/day), lorlatinib (100 mg/day), brigatinib (180 mg/day), alectinib (300 or 600 mg/day), ceritinib (750 mg/day), crizotinib (250 mg/day), and platinum-based chemotherapy  were considered. Phase III trials that included any of these agents were eligible for inclusion. Crizotinib was the first approved ALK inhibitor and the former first-line agent for initial therapy, and platinum-based chemotherapy was the first-line agent for ALK-p treatment-naive NSCLC prior to the approval of crizotinib. Thus, crizotinib or platinum-based chemotherapy was assumed as the common comparator for each treatment. The primary efficacy endpoint was PFS in all participants and in the subgroup of patients with CNS metastases, with corresponding HRs and 95% credible intervals (CrIs). To rank the relative efficacy of each therapeutic approach, the surface under the cumulative rank under the curve (SUCRA) values were calculated for each endpoint, with higher SUCRA values corresponding to a more preferred therapeutic approach for the corresponding endpoints . These aThe research for this systematic review and meta-analysis was a phase III trial of a parallel-group RCT.The Bayesian NMA was performed following robustly established methods developed at the National Institute of Medical Research ,109. We A sensitivity analysis  was condWe evaluated the statistical heterogeneity among the included studies to determine whether it impacts the final conclusions . StatistInstitutional Review Board approval and patient consent were waived because of the retrospective nature of this systematic review.This review outlines future treatment strategies and future prospects for ALK-p, ALK inhibitor-naïve advanced NSCLC with CNS metastasis, with a focus on elucidating and overcoming acquired resistance mechanisms. In addition, the therapeutic efficacies of ALK inhibitors in prolonging the PFS of the overall patients with ALK-p, ALK inhibitor-naïve advanced NSCLC and a subgroup of patients with CNS metastases are compared by drug and by generation. Generation-specific comparison shows that third-generation ALK inhibitors are significantly more efficient than second-generation and first-generation ALK inhibitors in prolonging the PFS of the overall patients and subgroup of patients with CNS metastases. Drug-specific comparison demonstrates that lorlatinib is the most efficient in prolonging the PFS of the overall patients and subgroup of patients with CNS metastases. Notably, although a significant difference in efficacy of prolonging PFS was found between lorlatinib and brigatinib in the overall patient population, no such significant difference was found in the subgroup of patients with CNS metastases. These results indicate a trend toward brigatinib as a promising first-line treatment option along with lorlatinib in the subgroup of patients with CNS metastases. These results can serve as a foundation for basic, clinical, and translational research and guide clinical oncologists in developing individualized treatment strategies for ALK-p ALK inhibitor-naïve advanced NSCLC. This NMA includes direct and indirect comparisons, and additional studies are warranted to confirm the results. The results of this analyses can serve as a basis for further clinical studies formulating novel treatment strategies for ALK-p ALK inhibitor-naïve advanced NSCLC with CNS metastases or acquired resistance mutations.


MedicDeepLabv3+ improves the state-of-the-art DeepLabv3+ with an advanced decoder, incorporating spatial attention layers and additional skip connections that, as we show in our experiments, lead to more precise segmentations. MedicDeepLabv3+ requires no MR image preprocessing, such as bias-field correction or registration to a template, produces segmentations in less than a second, and its GPU memory requirements can be adjusted based on the available resources. We optimized MedicDeepLabv3+ and six other state-of-the-art convolutional neural networks  on a heterogeneous training set comprised by MR volumes from 11 cohorts acquired at different lesion stages. Then, we evaluated the trained models and two approaches specifically designed for rodent MRI skull stripping (RATS and RBET) on a large dataset of 655 MR rat brain volumes. In our experiments, MedicDeepLabv3+ outperformed the other methods, yielding an average Dice coefficient of 0.952 and 0.944 in the brain and contralateral hemisphere regions. Additionally, we show that despite limiting the GPU memory and the training data, our MedicDeepLabv3+ also provided satisfactory segmentations. In conclusion, our method, publicly available at The online version contains supplementary material available at 10.1007/s12021-022-09607-1. Rodents are widely used in preclinical research to investigate brain diseases Carbone, . These sWith brain edema biomarkers in mind, our work focuses on cerebral hemisphere segmentation in MRI volumes of rat brains with ischemic lesions. Segmenting these images is particularly challenging since lesions’ size, shape, location, and contrast can vary even within images from the same cohort, hampering, as we show in our experiments, traditional segmentation methods. Additionally, rodents’ small size makes image acquisition sensitive to misalignments, potentially producing slices with asymmetric hemispheres and particularly affecting anisotropic data. Furthermore, although neuroanatomical segmentation tools can be used to produce hemisphere masks , these In recent years, convolutional neural networks (ConvNets) have been widely used to segment medical images due to their outstanding performance The T2-weighted MRI volumes were not preprocessed , and their intensity values were standardized to have zero mean and unit variance. Brain and contralateral hemisphere masks were annotated by several trained technicians employed by Charles River according to a standard operating procedure. These annotations did not include the cerebellum and the olfactory bulb. Finally, we computed the ipsilateral hemisphere mask by subtracting the contralateral hemisphere from the brain mask, yielding non-overlapping regions  for optimizing the ConvNets.We divided the MR images into a training set of 51 volumes, validation set of 17 volumes, and test set of 655 volumes. Specifically, we grouped the MR images by their cohort and acquisition time-point Fig. .Fig. 2MeMedicDeepLabv3+ is a 3D fully convolutional neural network (ConvNet) based on DeepLabv3+  is a quality metric that calculates the distance to the misclassification located the farthest from the boundary masks. Formally:A and B, respectively. In other words, HD provides the distance to the largest segmentation error. We provided HD values in mm and we accounted for voxel anisotropy. Finally, precision is the percentage of voxels accurately classified as brain/hemisphere, and recall is the percentage of brain/hemisphere voxels that were correctly identified:We assessed the automatic segmentations with Dice coefficient Dice, , HausdorWe compared our MedicDeepLabv3+ with DeepLabv3+ baseline  of MedicDeepLabv3+ at https://github.com/jmlipman/MedicDeepLabv3Plus.MedicDeepLabv3+, DeepLabv3+, UNet, HighRes3DNet, V-Net, VoxResNet, and Demon were implemented in Pytorch  and precision (0.94 and 0.94), and the lowest HD (1.856 and 2.064)  combining several preclinical neuroimaging studies to a large dataset of 723 rat MR volumes.ConvNets performed markedly better and their training time was about 10 times shorter than RATS Supplementary file2 (PDF 6.51 MB)Supplementary file3 (PDF 90.7 KB)Below is the link to the electronic supplementary material.


Children and adolescents with cerebral palsy (CP) are a vulnerable group who find it challenging to meet current physical activity guidelines, which predispose them to the negative health implications associated with low levels of physical activity and high levels of sedentary time. For these reasons, a key role for many clinicians, parents, and other practitioners working with children and adolescents with cerebral palsy is to encourage and facilitate an increase in habitual physical activity and reduce the amount of time spent sedentary, in order to optimize long-term health outcomes. Since 2014 Danish schools have been committed to enhance physical activity during the school day, but teachers still find it challenging to include children and adolescents with special needs. In Denmark, there is a strong tradition of practicing habitual exercise in the voluntary sports clubs (83% of children and adolescents). In sports clubs, these children are being physically active as well as experiencing being a part of a community. Children and adolescents living with CP are often not able to participate in these sports clubs which excludes them from the active and social life that's happening there. This study acknowledges that parents of children and adolescents living with CP play an important role in supporting them being physically active. Therefore, this study aims to identify perceived barriers and motivators for being physically active, experienced by this particular group of children and parents. This knowledge can be used by parents, clinicians, coaches, teachers and other practitioners to guide families living with CP towards a more physically active lifestyle and possibly optimize long-term physical and social health outcomes for children and adolescents with CP.This study will investigate the children's and parents' perceptions of motivators and barriers. The study is designed as a multi-family member interview study involving 10-14 combined in-depth interviews with children aged 8-15 (GMFCS I-III) and their parents. Interviews will be analysed thematically within and between groups.The study will take place in spring 2020 as a part of a pre-graduate research year and thus data and conclusions will be presented at the conference.


Their structures were elucidated through a detailed comprehensive spectroscopic analysis, as well as a comparison with the literature. An anti-inflammatory evaluation showed that compounds 2, 5, and 6 possessed inhibitory activity against the excessive production of nitric oxide (NO) and pro-inflammatory cytokines in LPS-treated RAW 264.7 macrophages in a dose-dependent manner without cytotoxicity. Further studies revealed that compound 2 was active in blocking the release of pro-inflammatory cytokines  induced by LPS both in vivo and in vitro. Our findings provide a basis for the further development of linear polyketides as promising anti-inflammatory agents.A new linear polyketide, named aspormisin A ( An acute inflammation response is usually triggered by infection or tissue injury, and the subsequent response is mainly mediated by tissue-resident macrophages and mast cells . A varieα and IL-1, have long been regarded as pro-inflammatory cytokines induced by LPS and have been determined as markers for the systemic activation of pro-inflammatory cytokines + . A detailed analysis of 1H NMR data ; three olefinic protons at δH 6.56 , 5.43 , and 5.15 ; two oxygenated methines at δH 4.87  and 3.74 ; two methines at δH 2.77–2.82  and 2.69–2.74 ; and seven methyls at δH 1.88 , 1.65 , 1.57 , 1.56 , 1.55 , 0.89 , and 0.78 . The 13C NMR data and HSQC spectrum displayed nineteen carbon signals, including one aldehyde carbon at δC 195.4 (C-1); an ester carbon at δC 169.4 (C-18); three olefinic quaternary carbons at δC 138.0 (C-2), 137.2 (C-6), and 132.8 (C-10); three olefinic carbons at δC 158.6 (C-3), 127.9 (C-7), and 123.3 (C-11); four methines (including two oxygenated) at δC 82.3 (C-9), 78.9 (C-5), 37.3 (C-4), and 33.7 (C-8); and seven methyls at δC 20.9 (C-19), 17.3 (C-16), 16.7 (C-14), 12.8 (C-15), 12.2 (C-12), 11.4 (C-17), and 9.1 (C-13). The sequential 1H-1H COSY correlations /CH-5, CH-7/CH-8(CH3-16)/CH-9, and C-11/CH3-12. The above NMR data indicated that the structural skeleton of 1 was similar to that of the co-isolated 5,9-dihydroxy-2,4,6,8,10-pentamethyldodeca-2,6,10-trienal (2) + , 667.4172 [2M + Na]+ .Aspormisin A -1223-336033 or e-mail: deposit@ccdc.cam.ac.uk).The clear light colorless crystal of 6: C10H10O4, Mr = 194.18, crystal size 0.16 × 0.14 × 0.10 mm3, monoclinic, a = 6.5123 (10) Å, b = 24.5467 (3) Å, c = 16.8673 (2) Å, α = 90°, β = 91.0470 (10)°, γ = 90°, V = 2695.88 (6) Å3, Z = 12, T = 100.00 (10) K, space group P21, μ(Cu Kα) = 0.944 mm−1, calcD = 1.435 g/cm3, 27695 reflections measured (5.24° ≤ 2Θ ≤ 148.846°), 10555 unique . The final R1 values were 0.0503 (I > 2σ(I)). The final wR(F2) values were 0.1342 (I > 2σ(I)). The final R1 values were 0.0538 . The final wR(F2) values were 0.1366 . The goodness of fit on F2 was 1.034. The flack parameter was 0.02(6). (CCDC 2132859).Crystal data for v/v) fetal bovine serum , penicillin (100 U/mL), and streptomycin (100 mg/mL)  with 5% CO2 at 37 °C. For LPS induction, RAW264.7 cells were seeded in a 24-well plate at a density of 1 × 105 cells/well, after adhesion cells were exposed to 0.1 μg/mL LPS  and separately co-treated with the compounds  for another 24 h. Culture supernatant was collected to measure the content of NO, IL-6, TNF-α, and MCP-1, and cells were harvested for RNA extraction.RAW264.7 cells, which originated from the American Type Culture Collection (ATCC) , were obtained from the Peking Union Medical College. Cells were cultured in Dulbecco’s modified Eagle’s medium (DMEM) with high glucose  supplemented with 10% (4 cells/well (100 μL medium/well) were seeded in a 96-well plate until adhesion; then, they were exposed to the compounds at a final concentration of 10 μM for 24 h, respectively. Whereafter, CCK-8 solution  was added following the manufacturer’s instructions and incubated at 37 °C for 1 h. The absorbance was read at 450 nm with a microplate reader . Cell viability was calculated by  × 100%.An amount of 3 × 10NO production was assessed with Nitric Oxide Assay Kit  according to the manufacturer’s instructions. Briefly, after 24 h of LPS treatment, the culture supernatant (50 μL) was transferred to another 96-well plate, and Griess reagents I (50 μL) as well as Griess reagents II (50 μL) were successively added. Absorbance was read at 540 nm.α , IL-6 , and MCP-1  in the culture supernatant collected above, according to the manufacturer’s instructions. Absorbance was measured at 450 nm.Mice enzyme-linked immunosorbent assay (ELISA) kits were used to measure the contents of TNF-® Reagent  and 1 μg total RNA was reverse transcribed into cDNA using EasyScript® One-Step gDNA Removal and cDNA Synthesis SuperMix . Gene transcript levels of IL-6, TNF- α, MCP-1, iNOS, Cox2, and MCP-1 were quantified using the TransStart Top Green qPCR SuperMix  according to the manufacturer’s protocol. β-actin was used as the housekeeper gene. Primers for quantitative PCR are shown in Total RNA was extracted from cells with TRIzoln = 6): the control group, the model group, and the compound 2 group. Mice in the compound 2 group were orally administrated with compound 2 (35 mg/kg) for 24 h before LPS (5 mg/kg) treatment, with saline used as the control in the other two groups. After 6 h of LPS administration, the animals were sacrificed for the collection of bronchoalveolar lavage fluid (BALF) and lung tissues. The content of inflammatory cytokines in BALF was determined by ELISA, and the upper left lungs from each group (n = 6 per group) were fixated with 4% buffered formalin solution. Serial sections 3 μm thick were stained with hematoxylin and eosin (H&E).All animal experiments have been approved by the Medical Ethics Committee of Peking Union Medical College  and comply with the regulations of the National Institutes of Health regarding the care and use of animals in research. Briefly, 6-week-old male BALB/c mice were purchased from Vital River Laboratories Co., Ltd. . The animals were randomly divided into 3 groups , were isolated from the alga-derived fungus A. ochraceopetaliformis SCSIO 41020. The X-ray single-crystal diffraction analysis of compound 6 was reported for the first time. Compounds 2, 5, and 6 dose-dependently inhibited the excessive production of NO and pro-inflammatory cytokines in LPS-treated RAW 264.7 macrophages without any evidence of cytotoxicity. The preliminary structure–activity relationship is also discussed. Moreover, the greatest potential inhibitor (2) further attenuated the lung injury in LPS-treated mice by downregulating the levels of pro-inflammatory cytokines, including IL-6, MCP-1, and TNF-α. Our findings provide a basis for the further development of linear polyketides as potential anti-inflammatory agents.In conclusion, six polyketides, including a new one (


Powered by this tool, coupled with domain expertise, we identify several potential COVID-19 drugs including methylprednisolone and metformin, and further discern key cellular SARS-CoV-2-affected pathways as potential druggable targets in COVID-19 pathogenesis.The current, rapidly diversifying pandemic has accelerated the need for efficient and effective identification of potential drug candidates for COVID-19. Knowledge on host-immune response to SARS-CoV-2 infection, however, remains limited with few drugs approved to date. Viable strategies and tools are rapidly arising to address this, especially with repurposing of existing drugs offering significant promise. Here we introduce a systems biology tool, the PHENotype SIMulator, which -by leveraging available transcriptomic and proteomic databases-allows modeling of SARS-CoV-2 infection in host cells   Soon afin vitro studies, in silico studies are of great value for rapid and effective drug discovery. Indeed, computational structure-based drug design and immuno-informatics have recently resulted in identification of potential SARS-CoV-2 target proteins and drugs that are being selected for further testing , target in Ref.  – recent in Ref. , however in Ref. .in silico with the addition of CD147 gene into KEGG, hereby developing an in silico ‘knock-in’ of the CD147 gene, in order to investigate the role of this extracellular matrix metalloproteinase inducer (EMMPRIN) in COVID-19  to the original simulator, if absent in KEGG. In the case of SARS-CoV-2 infection, the absence of some important genes involved in infection of host cells in the KEGG database, was considered a limitation. One gene in particular is Basigin (BSG)  gene. The importance of CD147 in SARS-CoV-2 infection in spike-protein (SP) binding and viral infiltration of host cells has been described . In our D-19 see .in silico model presented here provides an interesting framework that can be constinously developed and expanded further, achieving a more complete cell signature with input of (newly) available data on processes such as cell-cell communication through ligand-receptor complexes [in vitro studies are performed on cell lines, tissue tropism characteristics of viral infection seem key to better understanding viral activity [As discussed, the omplexes  or viralomplexes . As mostactivity . The samactivity ,62. Moreactivity . The sysactivity ,65. Inteactivity .Here we show distinct candidate drugs having a variable effect depending on the multiplicity of infection (MOI) of virus infection in A549-ACE2 expressing cells in low MOI 0.2; A and higin vitro SARS-CoV-2 drug screening [in vitro results  and identification of specific pathways of action of both pathogen and therapeutic compound in healthy and infected systems. For cost efficiency, validated predictive methods and assays for early elimination of potential drug candidates are of great value [in vitro experiments and clinical trials are time-consuming or impossible to reduce to practice. Optimally leveraging the power of pathway analysis by simulating host cell and tissue-specific infection and performing in silico drug selection, has a tremendous potential beyond COVID-19, with applicability to high global burden communicable diseases, translatable to pathogens of viral, bacterial and fungal origin, and potentially chronic disease such as inflamm-aging and diabetes. In conclusion, our PHENSIM approach will enable more effective in vitro experiments resulting in more rapidly initiated clinical trials and accelerated regulatory review of already pre-selected drugs with a high repurposable potential.PHENSIM gives rise to feasible validation and comparison of tal data ,9, givestal data ,52, tractal data , in siliat value . The oveat value , but in 4in vitro and cell-specific data, and will still need to be evaluated further in an in vitro and in vivo setting, as exemplified by HCQ which has not proven to be effective in vivo. Furthermore, our analysis is restricted to simulation of in vitro cell line data for specific cell types.The potential absence of some unknown important genes involved in SARS-CoV-2 infection of host cells in the KEGG databaseis a limitation of this study. Additionally, PHENSIM-determined potential candidates were based on 4Detailed methods are provided in the online version of this paper and include:-KEY RESOURCES TABLE-RESOURCE AVAILABILITYo Data and Code Availability:https://github.com/alaimos/phensim-covid19. Website: https://phensim.atlas.dmi.unict.it/[Bud Mishra: mishra@nyu.edu. Courant Institute of Mathematical Sciences Room 405, 251 Mercer Street, New York, NY 10012.o Lead contact: All input data, raw images, and source codes for PHENSIM are available atunict.it/.o Lead cConceptualization, design, drafting and revision of the manuscript: NIM, RVR, SA, BM, AJD, AF.Concept, design, and revisions of the manuscript: EB, AS, JACB, AP.Methodology: SA, AP; Writing original draft: NIM; Supervision: NIM, BM, AJD, AF.All authors declare no competing interests.https://github.com/alaimos/phensim-covid19. Website: https://phensim.atlas.dmi.unict.it/ [All input data, raw images, and source codes for PHENSIM are available at nict.it/ .Netherlands-Caribbean Foundation for Clinical Higher Education (NASKHO). S.A., A.F. and A.P. have been partially supported by the MIUR PON research project BILIGeCT “Liquid Biopsies for Cancer Clinical Management”.N.I.M. was funded in part by a fellowship award from the