Fluorinated graphene, an up‐rising member of the graphene family, combines a two‐dimensional layer‐structure, a wide bandgap, and high stability and attracts significant attention because of its unique nanostructure and carbon–fluorine bonds. Here, we give an extensive review of recent progress on synthetic methods and C–F bonding; additionally, we present the optical, electrical and electronic properties of fluorinated graphene and its electrochemical/biological applications. Fluorinated graphene exhibits various types of C–F bonds , tunable F/C ratios, and different configurations controlled by synthetic methods including direct fluorination and exfoliation methods. The relationship between the types/amounts of C–F bonds and specific properties, such as opened bandgap, high thermal and chemical stability, dispersibility, semiconducting/insulating nature, magnetic, self‐lubricating and mechanical properties and thermal conductivity, is discussed comprehensively. By optimizing the C–F bonding character and F/C ratios, fluorinated graphene can be utilized for energy conversion and storage devices, bioapplications, electrochemical sensors and amphiphobicity. Based on current progress, we propose potential problems of fluorinated graphene as well as the future challenge on the synthetic methods and C‐F bonding character. This review will provide guidance for controlling C–F bonds, developing fluorine‐related effects and promoting the application of fluorinated graphene. The chemical modification endows graphene with many excellent properties, such as good dispersion in organic/water solvents, chemical activity on the surface via functional groups and tunable electronic properties, such as bandgap opening, charge transfer density and work functions. Despite numerous studies, GO and halogenated graphene continue to have several problems as follows: (1) GO has a variety of chemical bonds containing carboxyl, carbonyl, hydroxyl, lactone, and epoxide at graphene edge and basal‐plan. The amount and concentration of these functional groups are not controlled. (2) Halogenated graphene usually is a mixture of nanosheets with different degrees of substitution and different halogen‐carbon bonds. (3) GO and halogenated graphene show less chemical and thermal stability than graphene as a result of a great amount of defects or substituents on the surface, especially for brominated‐ and iodine‐doped graphene. The representative characteristics of halogenated graphene were shown in TableGraphene oxide (GO)Fluorinated graphene is regarded as the two‐dimensional basic structural element of fluorinated graphite first synthesized by Ruff et al. in 1934.x, x ≈ 0–1.12), which is a stable and wide‐bandgap nanosheet in which a certain amount of C atoms is covalently bonded to F atoms, becomes a rising star of graphene derivatives because of its outstanding properties, such as a large negative magnetic resistance (a factor of 40 in a 9 T field), a wide optical bandgap (3.8 eV) and a high room‐temperature resistance (>10 GΩ).[[qv: 4a]],Fluorinated graphene (CF1.0 is 2162 Wh kg−1).x battery shows high energy densities, good chemical stability, a long‐term shelf life (>10 years) and minimal (<10%) self‐discharge.x (x = 1) is 865 mAh g−1 with an average discharge potential between 4.5 and 5 V for a purely ionic C–F bond.Compared with other derivatives, fluorinated graphene shows many unique properties because of the formation of various types of C‐F bonds. First, because the F atoms has a higher electronegativity (4.0) than C (2.5), H (2.2), and O atoms (3.4), fluorinated graphene show great potential for using as an atomically thin insulator or a tunnel barrier based on the heterostructure.[[qv: 4a]] Second, because of the difference in electronegativity (1.5) between C and F atoms, fluorinated graphene exhibits several C‐F bonding characters from ionic, semi‐ionic to covalent bonds controlled by the fluorination conditions.FigureRecently, much progress has been made on the preparation and control of C‐F bonding characters, F/C ratios (the F/C ratio is defined as the mole ratio of fluorine to carbon) and configurations of fluorinated graphene. The brief roadmap of a synthetic strategy of fluorinated graphene is shown in 2TableThe methods for synthesizing fluorinated graphene or fluorographene are mainly classified into two groups: fluorination and exfoliation methods. Fluorination mainly include direct gas‐fluorination,[[qv: 4a]],[[qv: 12a]],[[qv: 13b]],[[qv: 21a]],2.12.1.12 gas to treat a graphene film at 70 °C  on the Si substrate using XeF2 gas at room temperature.[[qv: 12a]] Fluorographene showed a dominant stoichiometry of C1.0F1.0 and a high F/C ratio of graphene film on both the front and back surface because of the effective etching on the Si substrate by XeF2 gas. This effect was confirmed by fluorinated graphene (CF0.25) on Cu foils because the Cu substrate cannot be etched by XeF2  is another important fluorination agent to prepare fluorinated graphene because of its high reactivity. Fluorinated graphene with different F/C ratios was synthesized by Wang et al.[[qv: 22a]] GO was treated by F2 at a low temperature (from room temperature to 180 °C) , which was treated using 1 atm F2 at a high temperature of 600 °C. The resultant few‐layer fluorinated graphene showed a high F/C ratio (CF0.7). Interestingly, Sofer et al.[[qv: 22e]] also presented an easy and weighable method for the fluorination of Hummers GO and Staudenmaier GO in 20% F2/N2 (v/v) at elevated temperatures and pressures. The high resolution XPS results indicated that the F/C ratio was 17.8% and 5.61% for Hummers GO and Staudenmaier GO, respectively.Fluorine gas F is anoth2 is limited by poor controllability of the C–F bonding characters (semi‐ionic or ionic bonds) and F/C ratios, special equipment requirements and environmental hazards (high toxicity and corrosion). Thus, many other fluorine‐containing agents such as SF6, SF4 or MoF6 were also used for fluorinating graphene. Pumera et al.[[qv: 22d]] demonstrated the fluorination of GO using SF6, SF4 or MoF6. The surface elemental composition showed that GO synthesized by the Hummers method were thermally fluorinated using SF6, SF4 and MoF6 at 800 °C with different F/C ratios of 1.92%, 0.53%, and 0.26%, respectively. Additionly, GO synthesized by Staudenmaier method were treated by microwave in SF6 at 800 °C and 1000 °C and showed diffeirent F/C ratios of 4.25% and 0.49%, respectively. The results revealed that F/C ratios of GO could be tuned by different gaseous fluorine‐containing agents with the control of the temperature for the fluorination. The structural changes of GO also led to the changes of F/C ratios under the treatment of SF6.[[qv: 22d]] Despite recent progress, the fluorination using XeF2, SF6, SF4 or MoF6 is still far from up‐scale industrial production. Thus, exploring a low‐toxic fluorine‐containing gas (or mixed gas) for mild, selective and high efficient fluorination is important for preparing various fluorianted graphene in the future.Despite the high activity, the fluorination of graphene using F2.1.26,] CF4,] and F2,[[qv: 23h]] have been used. Baraket et al.[[qv: 23a]] synthesized fluorinated graphene using electron‐beam generated plasmas in Ar/SF6  at room temperature.[[qv: 23b]] Yu et al.[[qv: 23k]] also synthesized fluorinated reduced graphene oxide (RGO) using CF4 plasma at room temperature, and the F/C ratios (F/C ≈ 0.17–0.27) were controlled by the plasma exposure time. Recently, Wang et al.[[qv: 23i]] reported the fluorination of CVD‐grown single‐layer graphene using CF4 plasma. The results showed that F/C ratios of fluorinated graphene were tuned by the conditions of the plasma; however, the resultant fluorinated graphene and fluorographene consisted of a mixture of CFx (x ≈ 1–3), and the spatial distribution of F on graphene was highly inhomogeneous.[[qv: 23i]] K. I. Ho et al.[[qv: 23e]] presented a one‐step approach for the selective fluorination of graphene using CF4 plasma in a plasma‐enhanced chemical vapor deposition (PECVD) system. During the fluorination, F‐radicals preferentially fluorinated graphene at a low temperature (<200 °C), while the defect was suppressed by screening out the effect of ion damage  energies are lower than fluoride radicals. Desipte a simple and effective method, the plasma fluorination inevitably damages the carbon structure of graphene by severe ion bombardment at a relatively high temperature.[[qv: 25a]],High‐density plasma is important for fluorinating graphene with high F/C ratios because the fluoride‐containing ions (such as F2.1.33‐etherate[[qv: 24b]] and diethylaminosulfur trifluoride (DAST)] and hexafluorophosphoric acid (HPF6).[[qv: 24d]]Hydrothermal or solvothermal fluorination is another versatile method for preparing fluorinated graphene and fluorographene. The fluorination effect depends on fluorine precursors, such as hydrofluoric acid (HF),[[qv: 24c]] BF3‐etherate and alkyl thiol/alkyl amine on the gram scale.Wang et al.[[qv: 24c]] presented a convenient method to fluorinate dispersed GO using HF through a simple hydrothermal process. Note that some oxygen‐containing groups were substituted by F atoms during the hydrothermal reaction. In addition, the F/C ratios were controlled by varying the temperature, times and HF concentration. Similarly, Gao et al.[[qv: 24a]] reported the solvothermal fluorination of GO films through converting the oxygen‐containing groups  to C‐F bonds by treating GO with DAST in chloroform at 50 °C. More recently, Samanta et al.[[qv: 24b]] prepared fluorinated RGO with fluorine coverage of 38 wt% using anhydrous BFGO is an excellent nanosheet for the hydrothermal fluorianation because of many epoxide, hydroxyl, carboxylic and ketone functional groups on the surface The oxygen‐containing groups can be removed or substituted by the formation of C‐F bonds at high temperature using a suitable fluorination solvent. Thus, the hydrothermal fluorination shows great potential for fluoriated graphene with high F/C ratios. Unfortunately, the uniform distribution of C‐F bonds on fluorinated graphene by the hydrothermal fluorination has yet been reported.2.1.42/Si substrate. Active fluorine radicals, generated by the decomposition of CYTOP, reacted with the sp2‐hybridized carbon and formed C‐F bonds  source and laser irradiation. The fluoropolymer CYTOP decomposed under laser irradiation on the surface of a single‐layer graphene film on a SiOs Figure d. Gong e2.22.2.12.G) of the intercalation compounding process triggered by the F atoms, which is defined in Equation H and ΔS are the enthalpy and entropy for the intercalation of molecules or solvents respectively. Because of van der Waals attraction between two adjacent layers of fluorinated graphite, ΔH is generally expected to be positive; resulting in a small and positive ΔG, and thus the exfoliation is mainly affected by TΔS. At high temperature and pressure, the increase in ΔS leads to a decrease in ΔG, which indicates increasing driving forces. Thus, compared with graphite, fluorinated graphene is easily exfoliated by the intercalation of molecules with relatively weak van der Waals attraction and a large interlayer space. High‐yield single‐ or few‐layer fluorinated graphene with a specific F/C ratio is obtained.Sonochemical exfoliation of multilayer materials has been well researched because it is a versatile and nondestructive technique for preparing high‐quality two‐dimensional single‐ or few‐layer nanomaterials. Solution‐processed exfoliation has been employed for up‐scale production of two‐dimensional graphene and MoSFigureZborˇil et al.[[qv: 9a]] prepared fluorinated graphene (F/C = 1.00) by a single‐step liquid‐phase exfoliation. In this process, fluorinated graphene was exfoliated from commercial fluorinated graphite suspended in sulfolane at a 135 W ultrasonic bath for 1 h at 50 °C. Chang et al.[[qv: 26a]] reported an effective and low‐cost exfoliation to obtain single and few‐layer fluorinated graphene (F/C = 0.25 or 0.50) in ionic liquid. In this method, ionic liquid (1‐butyl‐3‐methylimidazolium bromide) intercalated into the interlayer of commercial fluorinated graphite by mixing and incubating. After intercalation, black colloidal dispersion of fluorinated graphene was obtained by ultrasonication. Transmission electron microscopy (TEM) and atomic force microscopy (AFM) images revealed that two‐dimensional fluorinated graphene showed 1–5 layers with 2–10 μm in edge size. Among many organic solvents, NMP is considered to be an important intercalated molecule to exfoliate fluorinated graphene because of its dipole moment value of 4.09 D Figure4a.],3‐hybridized C atoms connect to F atoms and (2) fluorine radicals react with graphene to form semi‐ionic C‐F bonds, in which the sp2‐hybridized C atoms connect to F atoms. C–F bonds change from ionic to semi‐ionic to covalent, accompanied by a decrease in F/C ratios by changing the fluorination conditions .[[qv: 14a]] Borini et al.[[qv: 25b]] reported oxy‐fluorianted graphene with semi‐ionic C‐F bonds through the electrochemical intercalation of graphite in hydrofluoric acid solution. Wang et al.[[qv: 19b]] found that C‐F bonds showed the partial transformation from semi‐ionic nature to covalence with the increasing F/C ratios of fluorinated grapehene, which was controlled by the exposure time in XeF2 atmosphere. This transformation was also found in the liquid‐phase exfoliation with the appropriate solvent such as chloroform. Feng et al.[[qv: 26c]] found the partial transformation of covalent to semi‐ionic C‐F bonds in fluorinated graphene exfoliated by chloroform due to the formation of C–H…F hydrogen bonds between chloroform molecules and F atoms of fluorinated graphite.[[qv: 22a]],[[qv: 33b]],2‐F bonds.[[qv: 33b]] Previous studies indicated that the partial transformation between ionic (semi‐inoic) and covalent bonds could be caused by the interaction between C‐F bonds and other molecules or materials. Importantly, the natrue of C‐F bonds have a significant impact on the properties of fluorinated graphene, such as work function,[[qv: 23g]] reaction activity,The fluorination of C–C bonds of graphene usually contains two competing reaction processes: (1) fluorine radicals react with graphene to form covalent C‐F bonds, in which the sp−1 in the FTIR spectra.[[qv: 19b]],[[qv: 22a]],[[qv: 25c]],],TableThe presence and percentage of covalent, semi‐ionic or ionic C‐F bonds in fluorinated graphene are investigated by X‐ray photoelectron spectroscopy (XPS) and Fourier transform infrared spectroscopy (FTIR). According to previous studies, the characteristic peaks of semi‐ionic bonds between C and F atoms were observed at approximately 287–290 eV in the C1s XPS spectra Figure , 685–6883.2Precise control of the F/C ratio of fluorinated graphene is important for opening the bandgap, tuning electrical conductivity and optical transparency and understanding the structural transformation. Thus, beyond the aforementioned techniques in the second section, fluorination conditions, including the reaction temperature, the species of fluorination agents and catalysts, the type of carbon , the treated side and the sonochemical time, are utilized to tune the F/C ratios of fluorinated graphene.],[[qv: 24c]],[[qv: 26b]],4‐plasma treatment. Similar results were also observed in a recent study by Kuan‐I Ho et al.[[qv: 23e]] In addition, Wang et al.[[qv: 24c]] presented an easy, low‐cost and efficient hydrothermal‐process to tune F/C ratios of fluorinated graphene. The contents of each C‐F‐containing group  were dependent on the reaction temperature, time, and HF amount. An increase in the F/C ratios (from 0.11 to 0.48) was mainly attributed to the formation of the CF–CF2 group.[[qv: 24c]] Interestingly, Robinson et al. found that fluorine saturation coverage differed when graphene films were fluorinated by XeF2 on one or both sides. X‐ray photoelectron spectroscopy and Raman spectroscopy revealed that fluorine coverage saturates at 25% (C4F) for one‐side fluorination and at 100% (CF) for double‐side fluorination in XeF2 at room temperature.[[qv: 12a]] Gong et al.[[qv: 26b]] also reported that the F/C ratios decreased with increasing ultrasonication time in NMP, which might be attributed to the increasing stretching vibration energy of C–F groups gained from the sonic power facilitating the departure of fluorine.[[qv: 26b]]Yu et al.[[qv: 23k]] reported that F/C ratios (0.17–0.27) of fluorinated graphene were controlled by the time of CF3.3FigureFluorinated graphene and fluorographene consisting of weakly bound stacked two‐dimensional carbon monofluorides are a basic building block of fluorinated graphite.[[qv: 4a]],[[qv: 12a]],−1, which was twice the experimental value (100 ± 30 N m−1).[[qv: 4a]] The difference between the calculation and experiment might be attributed to a large number of structural defects in fluorographene because a certain portion of C atoms was not bonded to F atoms but formed dangling bonds.[[qv: 43a]] However, compared with fluorinated carbon nanotubes, the energy difference among various configurations of fluorographene is very small, and this result indicates that fluorographene is unlikely to be a pure single‐crystal form in a chair, stirrup, boat, or twist‐boat configuration.The first‐principles density functional theory (DFT) calculation showed that fluorographene with the chair configuration had a direct bandgap of 3.1 eV,4−1), good thermal stability below 400 °C, distinct nonlinear feature and high hydrophobicity. In this section, we discuss a variety of properties including bandgap, absorption or luminescence, stability, electronic conductivity, dspersibility, magnetic properties, tribological properties, mechanical or micromechanical properties, and thermal conductivity. These properties are significantly important for the application of fluorinated graphene.Fluorinated graphene shows many excellent properites such as wide bandgap of 3.1 eV, the highest theoretical specific capacity  using XeF2. The calculation indicated that the bandgap of fluorinated graphene increases with an increasing F/C ratio because of the interaction between the p‐orbital of F and the π‐orbital of C. The formation of sp3 bonds led to a large change in charge densities and scattering centers in the conduction band  showed a large bandgap of 3.07 eV.[[qv: 12a]] Liu et al.0.031, CF0.056, and CF0.125) were polar covalent bonds because of the high electronegativity of F atoms, and thus, they exhibited a metallic behavior. This behavior could be changed by increasing F/C ratios. CF0.25 and fluorinated graphene (CF1.0) had wide bandgaps of 2.92 eV and 3.13 eV, respectively, according to the generalized gradient approximation (GGA) calculations. Interestingly, fluorinated graphene (CF0.5) in which C atoms bonded to F atoms on one side also exhibited metallic behavior ascribed to the exchange splitting of the dangling C‐pz orbital with a coupling with an impurity state induced by F atoms. The results indicate that the bandgap of fluorinated graphene is greatly influenced by F/C ratios.Robinson et al.[[qv: 12a]] prepared fluorinated graphene films (on one side) with fluorine coverage of 25%  had a much narrower indirect bandgap than that of monolayer fluorinated graphene. Additionally, the bandgap of C4F nanosheets was further decreased by increasing the number of stacked layers because the conversion from insulator to semiconductor based on the dipole‐dipole interaction between two C4F layers induce a subtle interlayer polarization.Based on the density‐functional GGA calculation, the bandgap can also be controlled by different configurations and layers of fluorinated graphene and fluorographene. Specifically, a chair configuration shows a bandgap of 3.10 eV, while the bandgaps of the stirrup, boat, and twist‐boat configuration are 3.58, 3.28, and 3.05 eV, respectively.[[qv: 12b]] In the chair configuration, F atoms are alternately distributed on the plane. One F atom locates above the carbon layer, while the other one is under the same layer. Thus, the chair configuration has more symmetry than the stirrup configuration. In addition, the stirrup configuration is more significant for the conduction state because charge density follows the chain characteristic.[[qv: 12b]] Li et al.4.22. The absorption coefficient decreased after fluorination by SF6[[qv: 23c]] and CF4 plasma,[[qv: 23e]] which was in agreement with other fluorinated carbon materials.[[qv: 14b]],Fluorine‐substitution on carbon atom dramatically changes the optical properties of graphene including the absorption band, photoluminescence and transparency. Robinson et al.[[qv: 12a]] found that graphene film was optically transparent in the visible region after treatment by XeF2 at 70 °C  below the bandgap because of phonon‐assisted radiative recombination across the bandgap where the C–F vibration mode was excited when the electron‐hole pair recombined. Analogously, two accompanying peaks at 2.88 eV and 2.73 eV were also observed in low‐degree fluorinated graphene (fluorination for 1 day)  of fluorinated graphene have attracted attention because it not only yields insight into understanding electronic properties but is also crucial for advanced semiconductor devices and energy harvesting. Jeon et al.[[qv: 13a]] reported the room‐temperature PL spectra of graphene and fluorinated graphene dispersed in acetone using 290 nm (4.275 eV) excitation Figure d. The re[154–156] Samarakoon et al.[[qv: 12b]] reported the in‐plane absorption spectra of graphene and fluorographene calculated by GW‐BSE along with the random phase approximation (RPA) and GW‐RPA, respectively. RPA was regarded as the result of the DFT level. As shown in FigureOptical properties of ground‐state fluorinated graphene were also predicted by theoretical calculation based on DFT.154–1 Samarako4.3FigureD band. In contrast, fluorographene shows a high stability below 400 °C .[[qv: 4a]],[[qv: 12a]],[[qv: 22c]] The removal of both C and F atoms in fluorographene was observed by a prolonged annealing time at high temperature (≈450 °C).[[qv: 4a]] In addition, according to XPS data, fluorinated graphene, prepared using XeF2 gas on SiO2, Au, and Cu substrates, lost approximately 50–80% of the initial F/C ratios over 10 days until the F/C ratios were not changed.A below 200 °C. Fluorographene became weakly conductive, and the effective resistivity ρ = V/I decreased to ≈1 GΩ at 350 °C. The results indicated that the thermal stability and chemical inertness of fluorographene were similar to Teflon.[[qv: 4a]],[[qv: 13b]]Compared with the instability of GOn + nN2H4 → 4C + 4nHF + 2nN2. Radek et al.[[qv: 9a]] provided a pathway for defluorination using KI in DMF. In this process, fluorinated graphene transformed to metastable graphene iodide, which quickly decomposed to graphene and iodine at just 150 °C: CF+ KI → KF + [CI]; [CI] → C + 1/2I2. Additionally, Lee and co‐workers[[qv: 14d]] reported that ionic C‐F bond was selectively reduced by acetone treatment at a low temperature with the equation 2C2F(semi‐ionic) + CH3C(O)CH3(l) → HF + 2C(s) + C2F + CH3C(O)CH2(l).In addition, fluorographene showed a good chemical stability in many liquids such as water, acetone, and propanol, and under ambient conditions except for strong reductants.[[qv: 4a]],[[qv: 9a]] It was found that fluorinated graphene could be reduced by hydrazine, potassium iodide, ultraviolet irradiation and alkylamine compounds.[[qv: 9a]],[[qv: 12a]],More recently, new fluorinated graphene derivatives were prepared by the covalent modification of fluorinated graphene.4.42 hybridized C atoms with a pz orbital forming a π‐conjugated bond. Fluorination is widely used to chemically tailor the electrical conducitivity because it enables the transition from metallic/semiconducting to an insulating nature controlled by different F/C ratios.Single‐layer graphene shows high electron mobility because of its spI–V curves.[[qv: 4a]],[[qv: 22c]],[[qv: 23i]] Different from C–C bonds of graphene, every C atom in fluorographene with sp3 hybridization is bound to an F atom. Thus, fluorographene is an insulator because of the disappearance of π‐conjugated bonds. Wang et al.[[qv: 23i]] investigated the typical I–V characteristics of fluorinated graphene prepared by CF4 plasma. Fluorinated graphene showed a linear I–V curve with a short fluorination time (<10 min) with a resistance <10 MΩ because a low amount of F and sp3 C atoms were regarded as defects in an sp2 hybridized C network. Thus, the π‐conjugated network of graphene is preserved. However, this network was destroyed by a long fluorination‐time (>10 min) using CF4 plasma. Fluorinated graphene underwent the transition from semiconductor to insulator, resulting in a nonlinear curve with a resistance >1 GΩ. Compared with graphene, the resistance of fluorinated graphene showed a sharp increase by more than 7 orders of magnitude (from 10 kΩ to >100 GΩ) when the fluorine coverage is a few tenths of a percent.[[qv: 23i]],2 hybridized carbon network.−1, respectively.Fluorographene, the thinnest two‐dimensional insulator, shows a distinct nonlinear feature of 4.5z orbitals to form pseudohydrogen bonds with the hydroxy group of ethanol.[[qv: 13a]] The pseudohydrogen bond has been demonstrated to facilitate the dispersion of graphene with the π bond  in ethanol.[[qv: 13a]],[[qv: 26b]],Because of the presence of C‐F bonds, fluorinated graphene and fluorographene are highly hydrophobic and difficult to disperse or solubilize in most solvents because of its low surface free energy.[[qv: 21b]],z orbitals such as phenylethylene (PS), NMP, and THF than others with nonhybridized pz orbitals.[[qv: 26b]] Specifically, in a homogeneous solvent, a free pz orbital acted as an electron acceptor and formed pseudo‐hydrogen bonds with (CnF)x–F groups, thus resulting in an increase in dispersion of fluorinated graphene.[[qv: 13a]],[[qv: 26b]] Recently, the dispersion of fluorinated graphene in water was improved by fluorosurfactants in which perfluorinated units were adhered on the surface of fluorinated graphene and cationic or anionic units provided static repulsion.z orbitals, such as CH3CN, chloroform, and DMF.[[qv: 24e]]Gong et al.[[qv: 26b]] studied the dispersibility of fluorinated graphene in a variety of organic solvents. It was found that fluorinated graphene showed much better dispersion in solvents with a large closed conjugated system formed by p4.6T.x samlpes with x increasing from 0.1 to 1 was described by the Brillouin function. z = gJμBH/kBT, g was the g‐factor, J was the angular momentum number, N was the number of spins and kB was the Boltzmann constant. The number of spins N increased monotonically with x up to ≈0.9, and then showed some decrease for fluorographene. The maximum M achieved by the fluorination of graphene was one order of magnitude higher than that achieved by irradiation. Unfortunately, the concentration of magnetic moments was only ≈0.1% of the maximum hypothetically possible magnetism of one moment per carbon atom because F adatoms have a strongly towards clustering.[[qv: 73a]] Tang et al.[[qv: 73b]] reported that small F clusters that could be preferably formed around the vacancies in RGO produced a lot of magnetic edge adatoms. And such fluorinated RGO have a high magnetization of 0.83 emu g−1, a high magnetic moment of 3.187 × 10−3μB per carbon atom and a high efficiency of 8.68 × 10−3μB per F adatom. Recently, fluorinated GO was used as an efficient magnetic resonance imaging (MRI) contrast agent by Ajayan et al.Graphene obtained by sonochemical exfoliation of high‐purity HOPG shows a strongly diamagnetic response and no sign of ferromagnetism over a wide range of temperature, 4.73 configuration.[[qv: 76c]],[[qv: 77a]] Hou et al.Generally, graphene shows good tribological performance due to its high chemical inertness, extreme strength, easy shear capability on its densely packed and atomically smooth surface.4.84.8.1E) and intrinsic strength (σ) because of the presence of C‐F bonds. Nair et al. measured the E and σ of fluorographene using AFM.[[qv: 4a]] Fluorographene exhibited a lower E (100 ± 30 N m−1) and a lower σ (≈15 N m−1) than of graphene .[[qv: 4a]],E and σ arised from longer sp3 hybridized C–C bonds in fluorographene than sp3 hybridized C–C bonds in graphene.[[qv: 4a]] Interestingly, the elastic deformation σ/E of fluorgraphene showed litttle change in comparison of graphene because of the absence of structural defects during fluorination.[[qv: 4a]] The mechanism of controlling specifc mechanical properties including the strength, modules and deformation has yet been understood.Fluorination usually affects mechnical properities of graphene such as Young's modulus  were first used as the cathode in lithium primary batteries by Watanabe et al. in 1972.x was considered to be one of the ideal cathode materials for lithium primary batteries because of a variety of unique properties, such as high energy density, high average operating voltage, long shelf life, stable operation ability and wide operating temperature. Subsequently, Li/CFx batteries were first commercialized by Matsushita Electric Co. in Japan in 1975.x batteries have the highest theoretical specific capacity  in primary battery systems.[[qv: 14c]],[[qv: 18c]] With an ultrathin two‐dimensional layer‐structure, fluorinated graphene and fluorographene are regarded as the most promising CFx to achieve the theoretical capacity because of their tunable F/C ratios and C‐F bonding characters, favorable diffusion kinetics of lithium ions and large specific surface area. Recently, many studies focused on the performance of Li/CFx batteries using fluorinated graphene or fluorographene as the cathode material.[[qv: 17b]],],Fluorinated carbon materials , excellent rate capabilities (305 mA h g−1 at 5 A g−1), and outstanding cycling stability (capacity retention of ≈95% at 5 A g−1 after 2000 cycles) as the anode material for lithium ion batteries.−1 at 50 mA g−1 and excellent cycle performance for 50 cycles  (865 mAh g−1)  because of the high electronegativity of F atom (χ = 3.98) and the strong C‐F covalent bonds  at the edges.Feng et al. reported that lithium primary batteries using fluorographene exhibited a remarkable discharge rate because of good Lite Figure11a.],5.3Although fluorinated graphene is a semiconductor with a wide bandgap and shows UV‐fluorescence,[[qv: 12a]],[[qv: 13a]],5.4Fluorinated graphene can also be used for applications in supercapacitors,[[qv: 24e]] electrochemistry−1, which was much better than GO. The results were also confirmed by charge/discharge curves.[[qv: 24e]]Zhao et al.[[qv: 24e]] prepared solid supercapacitors using fluorinated graphene as an electrode material. Cyclic voltammetry measurements showed that fluorinated graphene prepared in dichloromethane exhibited the highest specific capacitance at 106.6 F gkobs0) at 2.69 × 10−3 cm s−1 and 4.37 × 10−3 cm s−1 in [Fe(CN)6]4–/3− and Eu2+/3+ redox probes, respectively. And the overpotentials of ascorbic acid and uric acid oxidations decrease with the increasing F/C ratios. And the fluorographite with F/C ratio of 0.75 provided a response to uric acid at 18.46 μA mM−1, which more sensitive than that to ascorbic acid (2.15 μA mM−1).Pumera et al.−1, synthesized by oxidizing the basal plane of fluorinated graphite. This method allows for unique, accessible, carbon‐based amphiphobic coatings.[[qv: 21b]]C‐F bonds drastically reduce the surface energy of graphene, resulting in a change in wetting behavior. Mathkar et al.[[qv: 21b]] reported an amphiphobic coating of fluorinated GO with a low surface tension of 59 dyn cm6In this review, we have given an overview of synthetic methods, structures and properties of fluorinated graphene that can be utilized for applications in high‐energy storage, unique biological response and magnetic resonance imaging, fluorinated graphene quantum dots, supercapacitors, electrochemistry and amphiphobicity. We have emphasized the importance and significance of controlling C‐F bonding characters, F/C ratios and configurations of fluorinated graphene, fluorographene and F‐GQDs by fluorination (gas or liquid phase) or exfoliation. The selective fluorination enables graphene with different two‐dimensional configurations for various properties including wide bandgap, blue luminescence, excellent electrochemistry, high stability and self‐lubricating. For example, an increase in the F/C ratio enlarges the bandgap of fluorographene, while a low F/C ratio usually ensures charge transport based on π‐conjugated structures. The covalent C‐F bonds in gas fluorination are crucial for thermal and chemical stability, while semi‐ionic and ionic bonds endow fluorinated graphene with a higher discharge potential for lithium batteries. Moreover, thermal conductivity, magnetic properties and luminescence of fluorinated graphene are not well developed because of a complicated fluoro‐carbon structure.Although significant progress has been made, additional challenge for uniform up‐scale synthesis/production, target‐oriented fluorination, the homogeneity of C‐F bonding character, solution processability and removal of other fluorides in applications must be addressed. It is very difficult to tune C‐F bonding precisely at the specifc microstructure and/or chemical structure because of the strong fluorination and the complicated chemical and microstructures  of graphene. A versatile, low‐cost and safe method of fluorinating graphene has not yet been found, resulting in the limitation of a wide range of use in commercial applications. Furthermore, because the mechanism of the formation of different C‐F bonding character is still unclear, fluorinated graphene is often composed of a mixture of various types of covalent, semi‐ionic and/or ionic C‐F bonds with different ratios. As a result, fluorinated graphene with different structures  needs to be separated and/or purified before the use in advanced electronic devices. Thus, chemical methods or strategies for selectively fluorinating graphene are of paramount importance.To date, there is a huge number of opportunities and challenges for designing and synthesizing fluorinated graphene with different structures such as core–shell, nanoporous spheres, nanocages and topologically nontrivial assemblies. The investigation of diffenent fluorinated graphene will put insightful understanding of their properties. On the basis of controlling the selectivity of plasma‐treatment, gas‐ and/or mild fluorniation, the change in F bonding nature, bandgap or electronic interaction of fluorinated graphene with the increasing F/C ratios and/or the specific C‐F bonds will be investigated. This result will further illustrate the effect of C‐F bonding character and configuration on thermal conductivity, self‐lubricating and optical properties.In addition to controllability and uniformity, multifunctional fluorinated graphene will be an interesting subject of intense and fruitful research in the future.A systematic study of thermal conductivity, magnetic property and luminscence will provide more special application in fluorinated graphene and F‐GQDs as well as other fluorinated carbon materials. Besides, the electrochemical properties of fluorinated graphene will be further improved to maxmize the energy density and power density by optimizing the microstructure, C‐F bonding character, the interfacial wettability and cooperation with additives. By the integration of chemical groups, polymer chains and/or functional nanoparticles, fluorinated graphene and its composites will show great potential for secondary batteries , super‐insulating materials, light emitting diodes (LED) and display materials. Based on much effort focused on the controllability of structures and optimized properties, in the future, fluorinated graphene will exhibit excellent performance in flexible nanoelectronics, energy conversion/storage, special protective coatings, and tissue engineering.


The as-synthesized samples were characterized by using field emission scanning electron microscopy (FE-SEM), high-resolution transmission electron microscopy (HRTEM), ultraviolet-visible (UV-vis) spectroscopy, Raman spectroscopy, X-ray diffraction (XRD), thermogravimetric-differential thermal analysis (TG-DTA) and gas-sensing test. The resistivity of the thick-film gas sensors based on sandwich-like graphene/WO3 nanocomposites can be controlled by varying the amount of graphene in the composites. Graphene/WO3 nanocomposites with graphene content higher than 1% show fast response, high selectivity and fine sensitivity to NOx.Graphene has attracted enormous attention owing to its extraordinary properties, while graphene-based nanocomposites hold promise for many applications. In this paper, we present a two-step exploitation method for preparation of graphene oxides and a facile solvothermal route for preparation of few-layer graphene nanosheets and graphene/WO Becau sensors ,14.3 is a well-known semiconductor material used for gas sensors to monitor toxic and explosive gases  [3 gas sensors is limited by poor selectivity, long response time, low sensitivity and high resistivity. Although methods, such as reduction of grain size, use of novel synthesis morphology, addition of dopants, and use of mixed sensing materials, have been adopted to enhance the sensing properties of WO3, fewer contributions have been made to reducing the high resistivity of WO3-based sensors [WOand NH3) ,18,19,20 sensors ,22.3 nanocomposites were prepared by a facile solvothermal method in an ethanol-distilled water medium. Material characterizations of the prepared samples were carried out by FE-SEM, HRTEM, UV-vis spectroscopy, Raman spectroscopy, XRD and TG-DTA. Gas-sensing properties of the sensors based on pure WO3, few-layer graphene nanosheets, and graphene/WO3 nanocomposites were investigated by using a HW-30A gas-sensing measurement system.In this paper, resistivity-controlled thick-film sensors based on few-layer graphene nanosheets and graphene/WOMorphologies of the first exploited graphite (EG) and graphite prepared from natural graphite by ultrasonication for 30 min are shown in −1, labeled as the D band, corresponds to breathing modes of rings or K-point phonons of A1g symmetry, whereas the G band at 1580 cm−1 corresponds to an E2g mode of graphite and is related to the vibration of sp2 bonded carbon atoms in a two-dimensional hexagonal lattice. The ratio of the intensity of the G band to that of the D band is related to the in-plane crystallite size, La. The in-plane crystallite size of FGO and FG, calculated by using the relationship La (nm) = 4.4 (I(G)/I(D)), about 4.51 and 3.7 nm, respectively, for which the corresponding I(G)/I(D) ratios are 1.0242 and 0.8315, respectively [−1, making it conceivable that the sample contained highly disordered and randomly arranged graphene sheets.Raman spectroscopy is a nondestructive and efficient approach to characterize graphitic materials, especially for determining ordered and disordered crystal structures of graphene. The Raman spectra of FGO and FG are shown in ectively . Compare3 nanocomposites can be indexed to hexagonal WO3(H2O)0.33 (JCPDS 87-1203), which is the same as the WO3 sample  to (002), which indicated that the preferred growth direction of WO3 was changed.XRD measurements were employed to investigate phase and structure of the synthesized samples. As shown in 3 sample d, and no3 and the FG (5%)/WO3 nanocomposites were further analyzed by FE-SEM and HRTEM. As shown in 3 sample exhibits unordered bundles with uniform strip size. TEM investigation can identify the bundled feature, giving evidence that each one-dimensional nanostructure bundle consists of a nanostrip with a thickness of about 100 nm, as shown in 3 nanocomposites. As displayed in 3 particles distributed randomly on the FG surface with different morphology are easily observed. The morphology change of WO3 indicates that the addition of FG has a significant effect on the WO3 particle growth, which is consistent with the XRD result, i.e., the preferred growth direction of WO3 was changed. The high-resolution TEM image of another area of the sample, given in 3(H2O)0.33. This is evidence for the existence of a sandwich-like structure between graphene and WO3. It is believed that both oxygen-containing defect sites and pristine regions of the GO favorably interact with W6+ via van der Waals interactions and chemisorptions, respectively. In addition, oxygen functional groups located at the surface of GO can act as anchor sites and effectively hinder diffusion, recrystallization, and growth of WO3 grains. The two morphologies of FG/WO3 nanocomposites may be formed via isolated growth of WO3 crystal species at the surface of GO and in situ growth of WO3 crystal species on the surface of GO, respectively.Morphologies of WO3 nanocomposites synthesized via the solvothermal method were characterized by TG-DTA, which was performed in air and N2 atmospheres with a heating rate of 15 °C·min−1. As shown in 2 atmosphere is significantly lower than that in air atmosphere especially at temperatures higher than 450 °C, which should result from the combustion of the carbon skeleton of graphene happened above 420 °C in air.Thermal properties and composition of the FG and FG (5%)/WO3 nanocomposites synthesized via the solvothermal method shows two abrupt weight losses occurring between 300 and 630 °C in air atmosphere. Correspondingly, the DTA curve shows an endothermic peak and an exothermal peak, which can be assigned to the loss of chemically bonded water and oxidation of graphene in air, respectively. As shown in 3 nanocomposites performed in N2 atmosphere exhibits a continuous weight loss, which can be assigned to the loss of chemically bonded water.From 3 sensors to different kinds of target gases were measured at different operating voltages. The results are shown in 3 sensors exhibit the largest response to NOx among all the tested gases at an operating voltage of 3.00 V. Furthermore, the optimal operating voltages are 3.00 V for NH3, 3.25 V for H2S and 4.00 V for acetone, dimethylbenzene, ethanol and trimethylamine. The response of the pure WO3 sensor to other target gases is extremely low at the tested operating voltage. As a result, the pure WO3 sensor is a very promising semiconductor for monitoring NOx at relatively low temperatures, but its high resistance limited its wider application.Selectivity is an important factor of gas sensors, so the responses of pure WO3-nanocomposite-based sensors with different amounts of FG  are shown in 3 nanocomposites with FG amount below 2%  is noticeably higher than that of the pure WO3 sensor . The sensors with few-layer graphene nanosheets and graphene/WO3 nanocomposites with graphene content higher than 1% have lower resistivity (below 135 kΩ) than that of the pure WO3 sensor. As we all know, WO3 is an n-type semiconductor and reduced graphene oxide is a p-type semiconductor, so it can form a p-n heterojunction between FG and WO3. The p-n heterojunction and the conduction of graphene play an important role in the resistance of FG/WO3 nanocomposites. As the amount of FG drops below 2%, the resistance of the FG/WO3 nanocomposites becomes controlled by the p-n heterojunction, so the resistance is really higher than that of WO3 and the sensors based on these are not suited for gas sensor application.Resistances of the FG/WO3 nanocomposites was studied at an operating voltage between 1.45 and 2.70 V. It was found that FG/WO3 nanocomposites only had gas sensitivity to NOx. The responses of FG/WO3 nanocomposite sensors with different amounts of FG to 100 ppm NOx at different operating voltages between 1.45 and 2.70 V are presented in x increased as the amount of FG decreased. Moreover, at operating voltages between 1.60 and 2.45 V, with the increase of operating voltage, the response of the FG/WO3 nanocomposite sensors was enhanced, and for operating voltages higher than 2.45 V, the response declined. As is well known, WO3 is very sensitive to NOx at relative higher operating voltage. As the operating voltage was increased to 2.45 V, the effect of WO3 on the response of the FG/WO3 nanocomposite sensors to NOx was enhanced. In other words, the counterbalance of the p-n heterojunction in the response of the FG/WO3 nanocomposite sensors to NOx cannot be ignored, so the response to NOx trends downward. Compared with the pure WO3 sensor, the sensitivity of FG/WO3 nanocomposite sensors was decreased, but both the resistance of the FG/WO3 nanocomposite sensors and the operating voltage were obviously lower.Gas sensitivity of the FG/WO3 nanocomposites, FG (3%)/WO3 nanocomposites, and FG (2%)/WO3 nanocomposites to 100 ppm NOx at an operating voltage of 2.45 V are shown in S = Rg/Ra, where Ra is a fixed value. The strong increase in Rg of the sensors upon exposure to 100 ppm NOx of the FG/WO3 nanocomposite sensors can be attributed to charge transfer between NOx molecules and the sensing material, where NOx act as an acceptor. It was clear that the response curves ascend or descend sharply with the inflow or outflow of NOx gas, indicating that the sensors have an excellent response-recovery property.The responses of FG, FG (5%)/WOAll the reagents used in the experiments were in analytic grade and used without further purification. Natural graphite, hydrogen peroxide, and ammonium persulfate were mixed at a weight ratio of 2:1:0.1 in a beaker at room temperature with ultrasonication for 5 min. Subsequently, the mixture was transferred to a crucible, and then it was placed in a microwave oven and irradiated at 500 W for 90 s . Under m2SO4 (98%). Then, KMnO4 (8.0 g) was added gradually under stirring and the temperature of the mixture was kept to below 10 °C by cooling. The reaction was maintained for 2 h below 10 °C. Then, the ice bath was removed, and the reaction mixture was stirred at 35 °C for 1 h, and diluted with 100 mL of distilled water. Because the addition of water in concentrated H2SO4 medium released a large amount of heat, the addition of water was performed to keep the temperature below 100 °C. After adding 100 mL of distilled water, the mixture was stirred for 1 h at 90 °C and further diluted to approximately 300 mL with distilled water. After that, 20 mL of 30% H2O2 was added to the mixture to reduce the residual KMnO4. Finally, the mixture was filtered and redispersed with 1 M HCl aqueous solution (300 mL) and left overnight. Several sets of filtering and redispersal processes were carried out to wash the SO42− completely. The resulting solid was dried at 50 °C for 24 h.Graphite oxides (GOs) were synthesized from EG using a modified Hummers method. In a typical procedure, 2.0 g of EG powder was put into 100 mL of cold (0 °C) concentrated H6) was dispersed in the mixture solution with continued stirring for 24 h. Then, the solution was transferred into a Teflon-line autoclave, which was put in an oven of 200 °C for 10 h. After the autoclave was naturally cooled to room temperature, the as-synthesized products were isolated by centrifugation, washed several times with deionized water and absolute ethanol to remove Cl−, respectively, and finally dried in a vacuum oven at 70 °C for 24 h. In addition, FG and WO3 were synthesized in the same way as the composites in the absence of WCl6 and GO, respectively.Graphite oxide was dispersed in 50 mL of distilled water with ultrasonication for 5 h (the second step of exfoliation of graphite), and the FGO solution was isolated by centrifugation, then 50 mL of ethanol was added into the FGO solution, and the pH value was adjusted to 8.0 with aqueous ammonia. One hundred and sixty mg of tungsten hexachloride  and a JEOL JEM-2100 (UHR) high-resolution transmission electron microscope (JEOL JEM-2100 (UHR) HRTEM). UV-vis spectroscopy measurements were performed on a UNICO UV-4802 UV-vis double-beam spectrophotometer  in ethanol dispersion. Raman spectra were recorded on a Renishaw inVia-Reflex confocal Raman microscope  with 532 nm laser excitation. Crystalline structure and crystallite size of the samples were characterized by x-ray diffraction , Cu Kα (λ = 0.15418 nm) radiation at 40 kV and 60 mA at room temperature. The TG-DTA measurement was carried out with a ZRY-1 instrument  at a heating rate of 15 °C/min with air and NS = Ra/Rg to the reducing gas and S = Rg/Ra to the oxidizing gas, where Ra and Rg are the resistances of a sensor in air and in a test gas, respectively.The fabrication process of gas sensors made from the as-prepared powder samples can be seen in . The gas3 nanocomposites were prepared via a solvothermal method in an ethanol-distilled water medium. It was found that the obtained few-layer graphene nanosheets have the intrinsic nature of graphene nanosheets. The preferred growth direction of WO3 was changed in the graphene/WO3 nanocomposites. The morphology of graphene/WO3 nanocomposites appeared as a sandwich-like structure. The few-layer graphene nanosheets and graphene/WO3 nanocomposite sensors with graphene content higher than 1% have obviously lower resistivity (below 135 kΩ) than that of a pure WO3 sensor , and they have the highest response to NOx at a relatively lower operating voltage (2.45 V).Graphene oxides were prepared by using a two-step exploitation method, and few-layer graphene nanosheets and graphene/WO


In addition, mutations at S409/410 of TDP-43 could inhibit its phosphorylation, attenuate nuclear loss, and abolish the increase in neuronal apoptosis in the subcortex. Inhibition of TDP-43 phosphorylation attenuated ICH-induced downregulation of mTOR activity and dynactin1 expression, which may relieve blocking of autophagosome-lysosome fusion and the increase of autophagosomal and lysosomal biogenesis induced by ICH. However, knockdown of TDP-43 could worsen ICH-induced SBI. Furthermore, TDP-43 could be dephosphorylated by calcineurin (CN), and CN activity was increased by OxyHb treatment. In conclusion, this study demonstrated that TDP-43 loss-of-function by phosphorylation at S409/410 may block autophagosome-lysosome fusion and induce elevation of LC3II and p62 levels by inhibiting the activity of mTOR and expression of dynactin1. This mechanism may play an important role in ICH-induced SBI, and TDP-43 may be a potential therapeutic target.This study aimed to determine the role of TAR DNA binding protein-43 (TDP-43) in intracerebral hemorrhage (ICH)-induced secondary brain injury (SBI) and its underlying mechanisms. After ICH, expression of TDP-43 in the nucleus was significantly decreased, and its expression in the cytoplasm increased both  Intracranial hemorrhage (ICH) is considered the most fatal subtype of stroke. It has a high mortality and accounts for 10%–15% of stroke cases  is a multifunctional DNA and RNA binding protein that participates in RNA transcription, alternative splicing, and regulation of mRNA stability in cells , phosphorylation, and ubiquitination. Phosphorylated TDP-43 plays important roles in formation of UPIs in FTLD and ALS  and increases autophagosomal and lysosomal biogenesis. In addition, loss of TDP-43 downregulates dynactin1 to impair fusion of autophagosomes with lysosomes  was provided by Novus Biologicals . Anti-mTOR (ab32028), anti-dynactin1 , anti-p62 (ab56416), anti-P-TDP43 (phospho S409 + S410), anti-NeuN (ab104224), and anti-histone H3 (ab1791) were purchased from Abcam . Anti-β-tubulin (sc-5274) was purchased from Santa Cruz .Adult male Sprague-Dawley rats weighing 280–300 g and pregnant rats were purchased from the Laboratory Animal Center, Medical College of Soochow University, Suzhou, Jiangsu, China. The animal experimental agreements were approved by the Experimental Animals Committee of the First Affiliated Hospital of Soochow University and complied with the ARRIVE guidelines. All rats were housed in a quiet environment maintained at 18–22°C with stable humidity and animals had free access to food and water. We strived to minimize the number of rats used and their pain.Rat (280–300 g) were fully anesthetized and then fixed in a stereotaxic apparatus frame . The rats were then placed supine on a heating pad that was maintained at approximately 27–35°C. The experimental ICH model was produced using stereotaxic insertion of a needle with a rounded tip and a side hole into the basal ganglia. The position of the basal ganglia was 3.5 mm lateral to the midline, 0.2 mm posterior to Bregma, and 5.5 mm ventral to the cortical surface. After the microinjector was in position, autologous blood was injected over 5 min, and the needle was left in place for an additional 5 min  and were randomly assigned to seven groups (six rats per group): sham group and six experimental groups arranged by time points of 3 h, 6 h, 12 h, 24 h, 48 h and 72 h after ICH. At the appropriate time point, rats were killed, and the right hemisphere basal ganglia tissues  were separated and collected for analysis . Neurons were maintained in a 5% CO2 incubator at 37°C. Half of the culture medium was replaced every 2 days for 1 week. Cells were then transfected with siRNA. To mimic ICH and evaluate effects of TDP-43 in vitro, cells were exposed to OxyHb at a concentration of 10 μM. After these treatments, cellular morphology was observed using an inverted phase contrast microscope, and total cell protein was collected and stored at −80°C until tested.Whole brains of 17-day-old rat embryos were used to prepare primary neuron-enriched cultures. We tried to minimize the number of embryos used and their suffering. In brief, we removed blood vessels and the meninges, and then the brains were digested with 0.25% trypsin for 5 min. Next, we centrifuged the brain suspension at 500× Nuclear protein extraction was performed using a Nuclear and Cytoplasmic Protein Extraction Kit  according to the manufacturer’s instructions.g for 10 min at 4°C. The supernatant was collected, and a standard BCA method  was used to determine protein concentration. Then, protein samples (100 mg/lane) were loaded onto a 10% SDS polyacrylamide gel, separated and electrophoretically transferred to a PVDF membrane . The membrane was then blocked with 5% nonfat milk for 2 h at 37°C. Next, the membrane was incubated with the primary antibody overnight at 4°C, followed by incubation with the horseradish peroxidase-linked secondary antibody for 1.5 h at 37°C. The membrane was washed with PBST and visualized using enhanced chemiluminescence detection . Relative quantities of protein levels were analyzed using ImageJ software.Brain tissues around the hematoma, which had been stored in liquid nitrogen, were thoroughly ground on ice. RIPA lysis buffer (Beyotime) was used to lyse the brain tissue for 30 min. The lysates were then transferred to a centrifuge tube and centrifuged at 12,000 We performed double labeling for TDP-43 and NeuN to assess expression of TDP-43 in neurons. The rat brain samples were fixed in 4% paraformaldehyde and embedded in paraffin. Next, sections were incubated with the primary antibody  overnight at 4°C followed by incubation with the NeuN antibody  overnight at 4°C. Then, sections were incubated with the secondary antibodies, which included Alexa Fluor 488 donkey anti-rabbit IgG antibody, Alexa Fluor 555 donkey anti-mouse IgG antibody, Alexa Fluor 488 donkey anti-mouse IgG antibody, and Alexa Fluor 555 donkey anti-rabbit IgG antibody . Normal rabbit IgG and normal mouse IgG were used as negative controls (data not shown). Finally, sections were observed using a fluorescence microscope , and relative fluorescence intensity was analyzed using ImageJ software.in vitro and detection by western blots. The most efficient siRNAs were used in this study, and the TDP-43 target sequences were as follows:Specific siRNAs against TDP-43 were provided by Ribobio. Knockdown efficiency of siRNAs was determined by transfection GAGAGGACTTGATCATTAACAGCGTGCATATATCCAATTGCTGAACCTAAGCATAATThe coding region of rat TDP-43 cDNA was subcloned into a pEGFP-N2 expression vector to produce the pEGFPN2-TDP-43 construct (without an EGFP tag). In addition, a rat TDP-43 cDNA construct with mutations at a possible key phosphorylation site  was also subcloned into a pEGFP-N2 expression vector (without an EGFP tag). All constructs were confirmed by DNA sequencing.in vivo RNA transfection reagent (18668-11-1 Engreen) according to the manufacturer’s instructions. Briefly, 5 nmol TDP-43 siRNA and 5 nmol scramble siRNA were dissolved in 66.5 μL DEPC RNase-free water. Then, 5 μL Entranster-in vivo RNA transfection reagents and 5 μL normal saline were added immediately to 10 μL siRNA or 10 μL scramble siRNA. The solution was mixed for 15 min at room temperature. Finally, 20 μL Entranster-siRNA mixture was injected intracerebroventricularly 48 h before ICH.Transfection of siRNA in the rat brain was done using Entranster-in vivo DNA transfection reagent (18668-11-2 Engreen) according to the manufacturer’s instructions. Briefly, 10 μL Entranster-in vivo DNA transfection reagent was added to 5 μg plasmid or 5 μg empty vector. The solution was mixed for 15 min at room temperature. Finally, 15 μL Entranster-in vivo-plasmid mixture was injected intracerebroventricularly 48 h before ICH.Transfection of plasmid in the rat brain was performed using Entranster-® 3000 Transfection Reagent  according to the manufacturer’s instructions. Neurons were transfected with TDP-43 siRNA, empty vector, TDP-43 plasmid, or scramble siRNA. Next, 48 h after transfection, neurons were exposed to 10 μM OxyHb for an additional 48 h. After the indicated treatments, neurons were harvested for further analysis.Transfection of siRNA and plasmid in cultured neurons was performed using LipofectamineBrain tissues embedded in paraffin were used for TUNEL staining. The sections were deparaffinized, dehydrated by heating at 75°C in an oven for 60 min, and then rehydrated through xylenes and graded ethanol solutions to water. The sections were then incubated in Triton X-100 for 10 min. After washing three times in PBS (5 min per wash), sections were incubated with the TUNEL reaction mixture for 1 h at 37°C. The sections were washed again three times in PBS (5 min per wash). After the final wash, sections were coverslipped with an anti-fading mounting medium containing DAPI. The number of TUNEL-positive neurons per millimeter was determined for each sample. Cell counts from each brain sample were averaged to provide a mean value.Cell necrosis in brain tissue was assessed using FJB. FJB procedures were identical to those for TUNEL. The sections were deparaffinized, dehydrated in an oven, rehydrated through xylenes and graded ethanol solutions to water, and permeabilized in 0.04% Triton X-100. The sections were then incubated in FJB dye solution and visualized using a fluorescence microscope . FJB-positive cells were counted by an observer who was blind to the experimental groups. To evaluate the extent of cell necrosis, the necrotic index was defined as the average number of FJB-positive cells in each section counted in six microscopic fields (400× magnification).Lactate dehydrogenase (LDH) activity in cell culture supernatants was quantified using a specific LDH assay according to the manufacturer’s instructions . First, reaction wells, which included standard wells, sample wells, control wells, and empty wells, were set up, and reagents sufficient for the number of assays to be performed were prepared. Then, a master mix of the reaction mix was prepared to ensure consistency of concentration and added to the relative groups. Finally, activity of LDH was measured immediately at OD 450 nm using a microplate reader.Analysis of CN activity was performed using a CN activity quantification kit  according to the manufacturer’s instructions.post hoc test. The former was used for comparisons to a single control group, and the latter was used to compare with the preselected groups. All data are expressed as mean ± SEM. A value of p < 0.05 was considered statistically significant.GraphPad Prism 6 was used for all statistical analysis. Data were analyzed by one-way ANOVA followed by either a Dunnett’s or a Sidak’s in vivo confirmed that TDP-43 translocated from the nucleus to the cytoplasm, and this shift peaked at 48 h after induction of ICH. The relative fluorescence intensity of TDP-43 was significantly increased at 48 h compared with the sham group, and then it rebounded at 72 h after ICH, which suggests that TDP-43 returns to the nucleus after 48 h  group than in the WT group Figures . Howeverin vivo experiments, we found that protein levels of mTOR, which reflect activity of mTORC1, were significantly decreased in the ICH groups compared with the sham group. Nevertheless, compared with the ICH group, mTOR levels were significantly elevated after TDP-43 overexpression, and the TDP-43 mutation group showed higher levels than the WT group. However, the TDP-43 knockdown group showed an obvious decreasing trend when compared with the Ctrl RNA group after ICH (Figures It has been reported that TDP-43 can participate in autophagosomal and lysosomal biogenesis as well as in autophagosome-lysosome fusion in some neurodegenerative diseases (She et al.,  Figures . Next, p Figures . Moreove Figures .in vitro, neurons were treated with 10 μM OxyHb. Immunofluorescence staining was used to assess TDP-43 expression in neurons after OxyHb treatment. After OxyHb treatment, TDP-43 translocated from the nucleus to the cytoplasm. Compared with the control group, the shift peaked at 48 h and then rebounded by 72 h after OxyHb treatment, which was similar to results of the in vivo experiments (Figures in vivo experiments, TDP-43 mutations could inhibit LDH release, in contrast to an increase in release with TDP-43 knockdown (Figure To mimic ICH  Figures . Furthern Figure .Previous studies have indicated that CN can dephosphorylate P-TDP-43 (Liachko et al., in vivo and in vitro, resulting in neuronal apoptosis and SBI. It is known that phosphorylation of TDP-43 affects its subcellular localization in part by formation of aggregates that prevent its return to the nucleus in both ALS and FTLD (Nonaka et al., Numerous recent studies have shown that phosphorylated TDP-43 plays an important role in formation of UPIs in FTLD and ALS (Fatima et al., in vivo and in vitro, and we found that levels of P-TDP-43 were significantly reduced after mutations at S409/410. Furthermore, these mutations reduced nuclear loss of TDP-43. In addition, with the mutations of TDP-43, ICH-induced SBI was ameliorated as shown by decreased neuronal necrosis and apoptosis as well as reduced LDH release. However, TDP-43 knockout increased ICH-induced SBI, which is similar to effects of TDP-43 in ALS, showing that neuronal loss of TDP-43 can induce FTLD (Yousef et al., There are multiple potential phosphorylation sites within TDP-43, and S409/410 plays the most critical role (Neumann et al., This study revealed roles of mTORC1 dysfunction as well as abnormal autophagy and lysosomal biogenesis in TDP-43-mediated neurodegeneration (She et al., Since CN can dephosphorylate P-TDP-43 (Liachko et al., In conclusion, this study provided new insight into the cellular mechanisms underlying TDP-43 nuclear loss-induced brain injury after ICH. Phosphorylation-induced TDP-43 translocation may enhance biogenesis of autophagosomes and lysosomes and partly block fusion of autophagosomes and lysosomes, resulting in ICH-induced SBI Figure . Thus, TGC and ZY contributed to the conception of this study and designed the research. LS and KZ were responsible for conducting the experiments about the animal model and writing the manuscript. WZ and HS conducted the experiments about the cell culture. HL and HS helped to design the study’s analytic strategy and perform the analysis with constructive discussion. All authors have critically revised the manuscript and approved the final 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.


PIK3CA-mutated GCs. A high PD-L1/PD-1 expression was associated with a significantly better patient outcome, and PD-L1 turned out to be an independent survival prognosticator. The correlation of PD-L1/PD-1 expression with distinct clinico-pathological patient characteristics may serve as a surrogate marker of PD-L1-positive GCs and may direct the use of immune checkpoint treatment strategies.Targeting the PD-1/PD-L1 immune checkpoint signaling is a novel promising treatment strategy in several tumor entities, and it is suggested that PD-L1/PD-1 expression is predictive for a PD-1/PD-L1 checkpoint inhibitor treatment response. We investigated the expression of PD-L1 and PD-1 by immunohistochemistry in a large and well characterized gastric cancer (GC) cohort of Caucasian patients, consisting of 465 GC samples and 15 corresponding liver metastases. Staining results were correlated with clinico-pathological characteristics and survival. PD-L1 expression was found in tumor cells of 140 GCs (30.1%) and 9 liver metastases (60%) respectively in immune cells of 411 GCs (88.4%) and 11 liver metastases (73.3%). PD-1 was expressed in tumor infiltrating lymphocytes in 250 GCs (53.8%) and in 11 liver metastases (73.3%). PD-L1 expression was significantly more prevalent in men, GCs of the proximal stomach, unclassified, papillary, Her2/neu-positive, Epstein-Barr-virus-positive, microsatellite instable, and  The genetic complexity of gastric cancer (GC) has been recently shown in an integrative genomic analysis including whole-genome sequencing. A molecular classification was proposed, which categorizes four subtypes, i.e. Epstein-Barr-virus (EBV)-positive, microsatellite instable (MSI), chromosomal instable, and genomically stable GCs , 2. ThesPD-L1 (B7-H1) is a 290aa type I transmembrane surface glycoprotein encoded by the CD274 gene located on chromosome 9 and expressed by several cell types of the immune system, e.g. lymphocytes and dendritic cells, but also aberrantly on the surface of epithelial cells of a wide range of solid tumors. PD-L1 is the ligand of programmed cell death 1 (PD-1), which is a member of the immunoglobulin superfamily B7 and involved in immunomodulation . PD-1 isCurrently, more than 400 studies worldwide focus on PD-L1/PD-1 immune checkpoint signaling, including 65 studies for cancer of the gastrointestinal tract , and theThe clinico-pathological characteristics of our patient cohort are summarized in Table PD-L1 expression was observed in tumor, stromal and immune cells, but not in non-neoplastic gastric epithelium. 140 of 465 cases (30.1%) showed a membranous PD-L1 expression in tumor cells. The percentage of stained tumor cells ranged from 0 to 80% (median 0%), the staining intensity ranged from 0 to 3  also serving as a positive control for the negatively rated tumor cell cases. The percentage of stained immune cells ranged from 0 to 70% (median 5%), the intensity ranged from 0 to 3 (median 1). As the majority of cases with PD-L1 positive immune cells showed a weak staining intensity , the intensity of the immunostaining was neglected for further statistical analyses, and only the percentage of PD-L1 positive immune cells was considered. All cases were graded into four groups as 0 (negative), 1 (1-9% positive), 2 (10-20% positive), and 3 (>20% positive). For further statistical analyses, group 0 and 1 were summarized and classified as “PD-L1 negative in immune cells” , and group 2 and 3 were classified “PD-L1 positive in immune cells” .The PD-L1 expression in stroma cells was mainly very limited to single cells, hence only the presence or absence was evaluated; PD-L1 positive stroma cells were observed in 107 of 465 cases  are known to be associated with PD-L1 overexpression , 15. NexMoreover, four different PD-L1 expression patterns were observed: (1) EBV-positive GCs often showed a heterogeneous, “patchy” expression pattern with a striking accumulation of PD-L1 positive tumor cells around larger blood vessels. (2) MSI-GCs were mainly PD-L1-positive at the interface between neoplastic and non-neoplastic tissue, especially in areas of “pushing borders”. (3) Papillary type GCs often showed PD-L1 positivity within the fibrovascular connective tissue cores and the intratumoral necrosis. (4) Other cases mainly showed no distinct PD-L1 distribution pattern and were classified as “patternless” .PD-1 expression was observed neither in tumor nor stroma cells. PD-1 positive diffusely distributed TILs were present in 250 of 465 cases (53.8%). As the amount of PD-1 positive TILs was mainly restricted to single cells, a quantitative analysis was neglected. Apart from a cumulative appearance of PD-1 positive immune cells in intratumoral lymphocyte aggregates/lymph follicles  or immune cells (group 2 and 3) showed a significant better overall and tumor specific survival Figure ; Table 1s Figure . There ws Figure .Patients with PD-1 positive TILs showed a better tumor specific survival, which was not significant after Simes' multiple testing procedure Table .in=0.05 and pout=0.10. These were UICC-stage, lymph node ratio, R-status, MET-status and PD-L1 expression in immune cells , and PD-1-status (immune cells). Five parameters remained in the Cox model after running the backward LR method with pls Table .Tissue specimens from liver metastases were available in 15 cases. Nine metastases (60%) showed a membranous PD-L1 expression of tumor cells, which was mainly limited to single cells and ranged from 0.1 to 3% respectively from 1+ to 2+ Figure . DichotoPD-L1 expression in immune cells was found in 11 metastases (73.3%) and ranged from 1 to 10% Figure . DichotoPD-1 positive TILs were present in 11 metastases (73.3%). The expression in GCs and their corresponding metastases was concordant in 12 cases  with a membranous PD-L1 expression in tumor cells, the tumor surface could not be evaluated. In the remaining 118 cases, PD-L1 positive tumor cells were close to the mucosal surface and generally accessible by biopsy in 79 (66.9%) cases. In 39 cases (33.1%), PD-L1 positive tumor cells were localized only in the tumor center or near the invasion front Figure .The present study is the first evaluation of PD-L1 and PD-1 expression in a large and well-characterized Caucasian cohort of GC. PD-L1 and PD-1 were expressed in a substantial amount of GCs either in tumor and immune cells or immune cells only. PD-L1 expression was found to be an independent survival prognosticator and correlated with distinct clinico-pathological patient characteristics.Up to now, only five studies of smaller Asian patient cohorts investigated the PD-L1/PD-1 expression in GC –16. PD-LAccurate interpretation of immunohistochemical stains is crucial for the establishment of a valid, histology-based predictive biomarker. Although we aimed for the HistoScore, we recognized that the overall percentage of PD-L1-positive tumor cells was low and the assessment of different percentages of three different staining intensities was indiscernible and impractical. Applying our simplified IRS, we next used patient survival as surrogate marker for biological relevance. Interestingly, while dichotomization into completely negative and any positive staining of tumor cells did not show a significant correlation with patient survival , dichotoA significant number of patients is diagnosed at an advanced, inoperable stage . Only biIn patients with a metastatic and/or unresectable GC, biopsy specimens might be obtained from the liver metastases, rather than the primary tumor, and biomarker expression between the primary GC and the liver metastasis might also show divergent test results. For Her2/neu, a discordance rate between 9 and 16% is known . In our In view of non-representative tissue sampling issues, we were next interested to test the hypothesis whether other clinico-pathological patient characteristics may serve as surrogate markers for PD-L1/PD1-expression. Correlation of PD-L1/PD-1 expression with various clinico-pathological patient characteristics demonstrated that PD-L1 is enriched in men, intestinal, unclassified or papillary type GC of the proximal stomach, EBV-positive, and MSI GCs. The latter findings confirm independently previous observations , 20. EBVPIK3CA-mutational status and combination of PD-L1/PD-1-targeted therapies with mTOR-inhibitors merit consideration in GC.The simultaneous application of separate monoclonal antibodies (mAbs) or multi-specific mAbs is still in its infancies but might offer new treatment strategies for complex diseases . Stagg ePD-L1 expression in GC of Western patients correlates significantly with overall and tumor specific survival as well as distinct clinico-pathological patient characteristics. Our findings might help to develop valid predictive test algorithms, which do not rely only on immunostaining patterns and may also help to explore novel combination therapies, e.g. trastuzumab or everolimus and PD-1/PD-L1 checkpoint inhibitors.All procedures followed were in accordance with the ethical standards of the responsible committee on human experimentation  and with the Helsinki Declaration of 1964 and later versions. Informed consent or substitute for it was obtained from all patients for being included in the study. Ethical approval was obtained from the local ethical review board (D 453/10).Epidemiological Cancer Registry of the state of Schleswig-Holstein, Germany. Follow-up data of those patients who were still alive were retrieved from hospital records and general practitioners. All patient data were pseudonymized prior to study inclusion.From the archive of the Institute of Pathology, University Hospital Kiel, we sought Caucasian patients who had undergone either total or partial gastrectomy for adenocarcinoma of the stomach or esophago-gastric junction between 1997 and 2009. The following patient characteristics were retrieved: type of surgery, age at diagnosis, gender, tumor size, tumor localization, tumor type, tumor grade, depth of invasion, residual tumor status, number of lymph nodes resected, and number of lymph nodes with metastases. Patients were included if an adenocarcinoma of the stomach or esophago-gastric junction was histologically confirmed. Exclusion criteria were defined as 1) histology identified a tumor type other than adenocarcinoma, and 2) patients had undergone a perioperative chemo- or radiotherapy. Each resected specimen had undergone gross sectioning and histological examination by trained and board certified surgical pathologists. Date of patient death was obtained from the th edition of the UICC guidelines [Tissue specimens were fixed in formalin and embedded in paraffin (FFPE). Deparaffinized sections were stained with hematoxylin and eosin. Histological re-examination of primary tissue sections was carried out for all cases to assure if inclusion criteria were met. Tumors were classified according to the Laurén classification  and re-eidelines .Immunostaining was carried out manually, using a rabbit monoclonal anti-PD-L1 antibody . Tissue sections were pretreated in citrate buffer for antigen retrieval and incubated with hydrogen peroxide block and Ultra V Block  to avoid unspecific reactions. For visualization the ImmPRESS-HRP-Universal–Antibody Polymer and the NovaRED substrate kit  were applied. Counterstaining was done with hematoxylin . Immunohistochemical PD-L1 stainings of liver metastases and PD-1 stainings of GCs and liver metastases were carried out with a Bondmax automated slide staining system , using the Polymer Refine Detection Kit  and the anti-PD-L1 antibody described above respectively a mouse monoclonal anti-PD-1-antibody . Germinal centers of lymph follicles served as internal positive control for both antibodies.Intensity and percentage of stained cells was evaluated separately for tumor, stroma and immune cells by two pathologists (CB and CR). For the evaluation of the PD-L1 expression in tumor cells, only the membranous staining was evaluated and the following immunoreactivity scoring system (IRS) was applied: Category A rated the percentage of immunoreactive cells and was graded as 0 (negative), 1 (≤1% positive), 2 (2 to 10% positive), 3 (11-50%) and 4 (>50%). Category B documented the intensity of immunostaining as 0 (no immunostaining), 1 (weak), 2 (moderate), or 3 (strong). The addition of category A and B resulted in an IRS ranging from 0 to 7. The evaluation of PD-L1 in stroma cells was rated as present or absent. For the evaluation of the PD-L1 expression in immune cells , only the percentage of positive cells was considered, and cases were graded as 0 (negative), 1 (1-5% positive), 2 (6-20%) and 3 (>20%). The immunostaining of PD-1 in immune cells was rated separately for tumor infiltrating lymphocytes (TILs) and intratumoral lymph follicles as present or absent.KRAS-, PIK3CA-, RHOA-, and GNAS-genotype was assessed as previously described [The mucin phenotype, Helicobacter pylori-, Her2/neu-, MET-, EBV-, MSI-status as well as the escribed , 26–29.Whole tissue sections from GCs and their liver metastases were stained with antibodies directed against PD-L1 and PD-1. The staining results were correlated with clinico-pathological and survival data.Statistical analyses were done using SPSS 20.0 . For the evaluation of PD-L1 in tumor cells and immune cells, marker expressions were first examined as raw score values and then dichotomized into positive and negative. Cross tabulations of clinical data and marker expressions were tested for independence using Fisher's exact test. The correlation between the PD-L1 and PD-1 expression in GCs and corresponding metastases was calculated by using Cohen's kappa. A kappa value of 0.20 was considered to be poor, of 0.21–0.40 to be fair, of 0.41–0.60 to be moderate, of 0.61–0.80 to be good, and of 0.81–1.00 to be very good. Median overall and tumour specific survival were calculated using the Kaplan-Meier method. Log-rank test was used to determine significance of differences between survival curves. Hazard ratios of variables were calculated by univariate Cox regression and those having p-values up to 0.1 were included in a multivariate Cox regression, combined with iterative backward LR method to identify independent prognostic variables. P-values up to 0.5 were considered as statistically significant. False discovery rate of correlations between clinical variables and biomarkers was controlled by applying the explorative Simes (Benjamini-Hochberg) procedure group-wise for each biomarker . P-value


This paper describes a study of the quantify surface roughness of experimentally manufactured particleboards and sandwiched panels having fibers on the surface layers. Surface quality of specimens before and after being overlaid with thin melamine impregnated papers was determined by employing profilometer equipment. Roughness measurements and Janka hardness were carried out on the specimens conditioned at 60% and 95% relative humidity levels. Based on the findings in this work, surface roughness of the specimens that were exposed two relative humidity exposure showed significant differences from each others. Data determined in this study could be beneficial to understand behavior of such panels exposed different humidity levels. In manufacture value-added products, wood based panels, such as fiberboards and particleboards, are used extensively as underlayment for thin overlays. These overlaid panels are main members in production of furniture and kitchen cabinets. As a well-known fact that wood-based composites are hygroscopic material and their dimensional stability varies with fluctuating as function of relative humidity. Especially, their surface quality depending on particle or fiber size on the face layer plays an important role on. Overall surface quality of panel is important so-called telegraphing effect of the roughness of the substrate shows through the overlay. Therefore, it is necessary to determine not only surface of the panel but also roughness of overlaid member with changing relative humidity so that they can be used more efficiency for different types of applications. Laminated and overlaid particleboard (PB) and medium density fiberboard (MDF) have been commonly used manufacturing indoor cabinets in Europa and the United States of America (USA) for over 35 years ,2. OverlIf surface irregularities on a composite panel are present, they may show through the overlays. Therefore, the overall quality of the final product can be affected by such irregularities on the substrate. In several previous studies, the stylus technique has been used to calculate the value of surface roughness of wood composites ,13,14,15In another previous work, medium density fiberboard surface roughness values were calculated using a fine stylus technique. During the sanding of the panels, surface roughness variations are introduced. In order to effectively evaluate this variation, previous studies used the profilometer technique . AlthougJuniperus virginiana L.) particles was supplied by a local sawmill in Oklahoma City. The particles contained both heartwood and softwood fractions of the trunk from eastern redcedar trees. Particles were dried to 2–3% moisture content in a laboratory type oven with a 1.0 m3 at the temperature of 67 ± 2 °C for 72 h. Dried particles were classified into two particle sizes, namely fine and coarse, on a 1 mm screen and 3 mm, respectively. After screening urea formaldehyde (UF) was blended with particles. Experimental panels were compressed at a temperature of 180 ± 2 °C and a pressure of 5.17 MPa for 5 min. All of the panels were pressed to a nominal thickness of 14 mm, and their target density was 0.70 g/cm3. Panels that were prepared with a length of 50 cm, width of 50 cm, and thickness of 14 mm. Manufacting process of composite panels is illustrated in Eastern redcedar (a), mean peak-to-valley height (Rz), and maximum roughness (Rmax), can be calculated from the digital information [Stylus method is a well accepted technique, resulting in quantitavie numerical values on the surface of sample. The profilometer consists of main unit and pick up which has a skid-type diamond stylus with 5 m tip radius and tracing span constant speed of 1 mm/s over 15.2 mm at a surface. Technical details and working principles of stylus type profilometer are presented in a past work . Roughneormation . a values of 3.02 μm. Samples with the size of 12 cm by 5 cm were used for random roughness measurements for test. A total of 20 samples were used for roughness measurements as illustrated in The calibration of the profilometer was checked every 100 measurements by using a standard reference plate with RHardness of the non-overlaid, overlaid particleboard, and non-overlaid, overlaid fiber sandwich panel specimens was tested by embedding a hemisphere steel having 11.2 mm diameter onto their tangential surface using a Comten 95 Series Universal Testing machine. Five measurements were taken from each sample and recorded in kg to evaluate their Janka hardness, as illustrated in 2. The test samples overlaid 50 s at Carver press with a 165 ± 5 °C under pressure of 2.3 MPa. Test samples were conditioned in a chamber with a temperature of 20 °C and a relative humidity of 65% until they reach to the equilibrium moisture content before any roughness measurements were taken from their surface. The overlaying process of the specimens is shown in A total of ten, five for each type of composites with dimensions of 12 cm by 5 cm were overlaid with melamine based decorative paper having weight of 7000 g/cmAfter initial measurements were taken from surface of samples, they were placed in chamber having 95% relative humidity and were kept for 10 days. In the next step individual samples were weighted at an accuracy of 0.1 g. Later roughness measurements from the surface of each overlaid samples. Roughness values of the samples were quantitatively evaluated at initial dry condition and as they were exposed to relative humidity levels of 60% and 95%. Penetration of humidity on roughness of overlaid and non-overlaid substrate is schematically illustrated in In order to analyze the significant differences of all the parameters that were used in this study, one-way analysis of variance (one-way ANOVA) was used. All results were computed employing IBM Statistical Package for the Social Sciences software (SPSS), version 21 .a value of 0.48 μm once they were exposed to 60% and 95% relative humidity levels, corresponding values were 0.62 μm and 1.67 μm, respectively. Measurements of average surface roughness values of fiber-sandwich composite panels are illustrated in z and Rmax as well.As expected, relative humidity effects all three measures of surface roughness in both fiber sandwich boards. It is seen in a values once they were exposed to 60% and 95% relative humidity levels, as illustrated in It seems that PB samples had rougher initial average surface values, however they had similar trend having higher RIn p < 0.05) on interaction effects between panel types and humidity level. Based on ANOVA, significant difference was observed between main effects such as composite panel types and humidity levels at a 95% confidence level. The average surface roughness values were found to be effective (After relative humidity exposure process, the surface roughness of composite panel specimens increased from 3.22 μm to 5.82 μm and 80% increase in the surface roughness was found. Both fiber-sandwich type and particleboard samples had higher surface roughness values at 95% humidity level. It appears that humidity level exposure effected surface quality of fiber-sandwich panels. In a had the greatest increase with a value of 83%, followed by Rmax 64%, and finally Rz the least 38% increase. If the trends are compared between the surface roughness characteristics, Ra had a steeper slope than that of other parameters. Therefore, it appears that an increase in relative humidity adversely influenced overall surface quality of the samples. It can be noticed that as percentage of relative humidity increase, all of the surface roughness values increase. Specifically, RIncreasing the percentage of relative humidity increased all the parameters of the surface roughness; this difference is due to the overlay, which protects the surfaces against relative humidity. Using overlay on the fiber sandwich board surfaces reduces the negative effects of relative humidity with an approximate value of nine times. Both MDF-Sandwich panel samples had a higher surface roughness values at 95% humidity level. Humidity levels effected MDF sandwich panels directly and surface quality deteriorated with humidity. In a had the greatest increase with a value of 58%, followed by Rmax 30%, and finally Rz, with the least 8% increase. If the trends are compared between the surface roughness characteristics, Ra had a steeper slope than that of other parameters. Therefore, it seems that increase in relative humidity adversely influenced the overall surface quality of the samples.The percentage of relative humidity increase, all surface roughness values of non-overlaid particleboard increase. Specifically, Ra had the greatest increase with a value of 143%, followed by Rmax 96%, and finally, Rz the least, with an 86% increase. If the trends are compared between the surface roughness characteristics, Ra had a steeper slope than that of other parameters. Therefore, it appears that the increase in relative humidity adversely influenced all the measures of surface quality for each sample evaluated. Increasing the percentage of relative humidity increased all parameters of the surface roughness, this difference is due to the overlay, which protects the surfaces against relative humidity. Using overlay on the particleboard surfaces reduces the negative effects of relative humidity, with an approximate value of six times. All of the surface roughness values of overlaid particleboard increase after increasing relative humidity. Specifically, RAs percentage of relative humidity increase, all Janka hardness values decrease. Specifically, non-overlaid particleboard had the minimum Janka harness value 171 kg overall. If the Janka hardness values are compared between the overlaid and non-overlaid specimens, overlaid samples have better Janka hardness values between 5% and 6%. Therefore, it appears that the increase in relative humidity adversely influenced the overall hardness values of the specimens. This study aimed to quantify the effect of humidity exposure on both surface quality and Janka hardness for different types of wood composite material, namely these were overlaid and non-overlaid composite panels. Based on the findings that are discussed in this work, as a conclusion, overlaid samples that were exposed to different levels of relative humidity had less overall surface roughness characteristics than the non-overlaid samples. Furthermore, the Janka hardness results were higher in the case of overlaid samples than in the non-overlaid samples. The data found in this work can inform future decisions to make more efficient use of overlaid and non-overlaid wooden composite boards. This work can be extended in further studies by evaluating other mechanical properties, such as bending and compression strength, of wood composite boards under different levels of relative humidity. Additionally, through future studies of dimensional stability, as effected by relative humidity, can gain a better understanding of the behavior of overlaid panels.


Autophagy is a catabolic mechanism where intracellular material is degraded by vesicular structures called autophagolysosomes. Autophagy is necessary to maintain the normal function of the central nervous system (CNS), avoiding the accumulation of misfolded and aggregated proteins. Consistently, impaired autophagy has been associated with the pathogenesis of various neurodegenerative diseases. The proteins TAR DNA-binding protein-43 (TDP-43), which regulates RNA processing at different levels, and chromosome 9 open reading frame 72 (C9orf72), probably involved in membrane trafficking, are crucial in the development of neurodegenerative diseases such as Amyotrophic lateral sclerosis (ALS) and Frontotemporal Lobar Degeneration (FTLD). Additionally, recent studies have identified a role for these proteins in the control of autophagy. In this manuscript, we review what is known regarding the autophagic mechanism and discuss the involvement of TDP-43 and C9orf72 in autophagy and their impact on neurodegenerative diseases. The population is aging worldwide. Indeed, between 2015 and 2030, the number of people in the world aged 60 years or over is projected to grow by 56 per cent, from 901 million to 1.4 billion; by 2050, the global population of older persons is projected to more than double in size, reaching nearly 2.1 billionNeurodegenerative diseases such as Alzheimer's, Parkinson's, Prion, Huntington's, and Motor neuron diseases (MND), as well as Spinal muscular atrophy, Spinocerebellar ataxia (SCA), Frontotemporal Lobar Degeneration (FTLD), and Amyotrophic lateral sclerosis (ALS) cause progressive damage in the central and peripheral nervous system, reducing the quality of life of affected patients , a form of dementia characterized by a progressive decline in behavior and/or language  genes are involved in the process of autophagic vacuole formation  in membrane microdomains where proteins with the PtdIns3P-binding (FYVE) and the conserved Phox-homology (PX) domains are recruited to form a protein elongation platform at the omegasome , which are considered cellular nutritional sensors able to modulate different signaling pathways  at Thr1227, which, together with TSC1, forms the TSC complex  is another negative regulator of autophagy  inhibits autophagy and promotes a neurodegenerative phenotype characterized by axonal degeneration and accumulation of aggregate-prone proteins in the cytosol of neurons without altering proteasome function  the gene encoding for Beclin 1 shows decreased mRNA levels in brain tissue, suggesting reduced autophagosome formation and increased development of AD , where omegasomes formation is inhibited due to the mislocalization of the protein ATG9, preventing the formation of new autophagosomes. This reduces the clearance of the protein α-synuclein, which accumulates in intra-neuronal inclusions called Lewy bodies, a feature of the brain tissue of PD patients . Autophagosomes move along microtubules in the direction of the perinuclear microtubule-organizing center of the neuron, which is enriched in lysosomes . Inefficient autophagosomal clearance can be associated with reduced lysosomal acidification, which reduces the activity of lysosomal hydrolases and promotes the accumulation of undigested byproducts. This is typical of the so-called “lysosomal storage disorders” (LSD), which comprise more than 50 diseases and represent the most common forms of neurodegenerative disease in childhood. While the proteins and mechanisms involved in these diseases differ, impaired autophagy is unfailingly present , hereafter called TDP-43, was initially described as a transcriptional repressor factor of HIV n]. These repeats locate in a non-coding region of C9orf72, specifically within the intron that separates exons 1a and 1b , whereas affected individuals incorporate between 30 and 250 hexanucleotide repeats (n ≥ 30) n repeats may involve both, a reduction in the expression of some C9orf72 variants and/or the aberrant formation of RNA foci n repeats, in all reading frames, and in the absence of an initiating ATG codon. Thus, this mechanism can generate polypeptides formed by different numbers of dipeptide units originated from one (G4C2)n repeat. These polypetides, known as DRPs (Dipeptide Repeat Proteins) or C9RANT proteins, have shown to be toxic for the cell, since they impair RNA biogenesis n repeats have been recently reviewed by Todd and Petrucelli  and CHMP2B. OPTN, which is involved in the mitophagy machinery, is proposed to impact on ALS as its mutations promote severe mitochondria degradation  as a new gene associated with ALS  and OPTN, which co-localize with TDP-43, p62/SQSTM1 and ubiquitin in spinal motor neurons of sporadic ALS patients . Indeed, it has been observed that HSP-90, cell division cycle 37 (CDC37) and TDP-43 form a protein complex that prevents TDP-43 degradation in HeLa cells. The disruption of this complex, by downregulation of HSP-90 or CDC37 promoted degradation of TDP-43 via induction of autophagy  are strictly coordinated to degrade WT and aggregated forms of TDP-43  , a master transcription factor that regulates the expression of genes controlling lysosomes biogenesis and autophagy induction , TBK1 Binding Protein 1 (SINTBAD) and Nucleosome Assembly Protein 1 (NAP1), forming a protein complex that activates the TANK-Binding Kinase 1 (TBK1). Active TBK1 phosphorylates SMCR8 at Ser 402 and Thr 796, stabilizing the C9orf72/SMCR8 complex and thus inducing autophagy. On the other hand, the interaction between C9orf72/SMCR8/WDR41 and ULK complex induced SMCR8 phosphorylation at Ser 400, 492, 562, and Thr 666/Ser 667. These modifications are less efficient in inducing autophagy compared with those performed by TBK1 (Sellier et al., in vivo model, the over-expression of Ataxin-2 Q30 aggregates induced cell toxicity when autophagy was inhibited by C9orf72 down-regulation (Sellier et al., In agreement with the role of C9orf72 in autophagy, different works have demonstrated that C9orf72 down-regulation impaired autophagy progression, as evidenced by the intracellular accumulation of p62/SQSTM1 (Farg et al., Autophagy dysfunction has been identified in several neurodegenerative diseases such as ALS and FTLD. TDP-43 and C9orf72 are also associated with these illnesses as well as other neurodegenerative disorders. TDP-43 aggregates are found in neurons of ALS and FTLD patients, and it has been proposed that they may act as a sink for the newly synthetized TDP-43 protein, thus leading to TDP-43 loss-of-function. Autophagy has been proposed to be crucial for the maintenance of TDP-43 protein levels and to avoid TDP-43 aggregation. Indeed, very recently, data indicate TDP-43 is able to regulate autophagy by stabilizing the mRNA of important components of the autophagic machinery, and, conversely, TDP-43 down-regulation/loss-of-function finally inhibits the autophagic flux.The association of C9orf72 with ALS and FTLD has recently been discovered, and the physiological functions of C9orf72 are still being investigated. Additionally, recent studies showed that C9orf72 is involved in membrane trafficking and autophagy. Specifically, it has been demonstrated that C9orf72 stimulates autophagy initiation by interacting with autophagic proteins involved in membrane trafficking. Consistently, in diseases such as ALS and FTLD, it has been observed a reduction in C9orf72 as well as decreased levels of autophagy.Thus, to date, evidences strongly support that both, TDP-43 and C9orf72 regulate autophagy and that loss-of-function of any of these factors blocks the autophagic flux. In the scenario of TDP-43-associated proteinopathies, TDP-43 loss-of-function caused by TDP-43 aggregates, can further increase cellular stress by blocking the autophagic process. In turn, autophagy inhibition provokes an exacerbated accumulation of TDP-43 aggregates, enhances cellular stress and induces cell death, leading to the onset or progression of neurodegenerative diseases.Consistently, in C9orf72-associated neuropathologies, two events lead to the onset of the disease. The first one is associated with the production of aberrant protein inclusions, such as TDP-43 or Ataxin aggregates, while the second one is characterized by the deregulation of autophagy caused by C9orf72 loss-of-function.Thus, the dysregulation of autophagy is key in the development of TDP-43- and C9orf72-associated neurodegenerative disorders. Even though different are the mechanisms that cause the impairment in autophagy, induction in the autophagic pathway can be considered as a general strategy to avoid these diseases, and research should focus on the identification of novel therapeutic strategies to stimulate autophagy.MB, EM, and AC wrote the article and made the figures. EB made important contributions, discussions and corrections to the manuscript.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.