With highly nonplanar topological configuration, TPA(PDAF)3 shows weaker intermolecular interaction in the solid states and thus exhibits single nanomolecular behavior, which is crucial for charge stored and retained in an organic field‐effect transistor (OFET) memory device. Furthermore, diazafluorene derivatives possess a completely separate highest occupied molecular orbital/lowest unoccupied molecular orbital, which offers ideal hole and electron trapping sites. As charge storage elements, triphenylamine groups provide the hole trapping sites, while diazafluorene units provide the electron trapping sites and act as a hole blocking group to restrain the leakage of stored holes trapped in triphenylamine. The pentacene‐based OFET memory device with solution‐processing TPA(PDAF)3 shows a good hole‐trapping ability, high hole trapping density (4.55 × 1012 cm−2), fast trapping speed (<20 ms), a large memory window (89 V), and a tunable ambipolar memory behavior. The optimized device shows a large ON/OFF current ratio (2.85 × 107), good charge retention (>104 s), and reliable endurance properties. This study suggests that diazafluorene based donor–acceptor small molecular elements have great promise for high‐performance OFET memory.Three diazafluorene derivatives triphenylamine (TPA)(PDAF) Among the reported polymer electrets, polystyrene (PS)‐based side‐chain polymers] and donor–accepter polyimides1, TPA(PDAF)2, and TPA(PDAF)3 were prepared to explore the role of the electronic structure and charge trapping mechanism. Herein, the preparations, characterizations, spectra, electrical properties, and theory calculation of TPA(PDAF)n were systematically explored. It is worth mentioning that the TPA(PDAF)n elements were prepared by simple solution‐processing methods instead of vacuum evaporation like most small molecules reported.] The charge‐trapping and ambipolar memory characteristics of pentacene‐based OFET memory using TPA(PDAF)n as charge storage layers were studied. In addition, the relationships between electronic structure of TPA(PDAF)n, charge trapping, and memory characteristics was discussed in detail.In order to achieve efficient small molecular elements, in this study, donor–acceptor type molecules with triphenylamine (TPA) and 4,5‐diazafluorene units were designed and synthesized to serve as charge storage layers in OFET memory devices. As a classic hole‐transporting moiety, TPA unit has been proved to be an efficient hole trapping core in OFET memory.22.1n  are listed in Scheme1 was obtained at −78 °C, while by‐products tertiary alcohols 2 and 3 were synthesized at higher temperature. Therefore, route I was thermodynamic‐controlled grignard reaction. Subsequently, TPA(PDAF)n  was smoothly prepared by 1 and TPA via concentrated sulfuric acid‐mediated Friedel–Crafts reaction with the yields of 56%, 46%, and 75%, respectively. Chemical structures of the products were confirmed by 1H and 13C NMR, high‐resolution mass spectrometry (HRMS), and fortunately, the single crystals of compounds 3 and 6 were also obtained. The data could be seen in the Supporting Information. The single crystals of compounds 3 and 6 were unambiguously characterized by X‐ray crystallographic analysis, as shown in Figure S1 (Supporting Information). The single crystal of TPA(PDAF)3 revealed a 3D topology shape, consisting a core unit of TPA and three open‐arms of 4,5‐diazafluorene moieties. Owing to the large steric hindrance of diazafluorene moieties connecting to TPA, TPA(PDAF)3 displayed the asymmetric conformation in three directions with open‐armed at angles of 122°, 113°, and 125°. Compared with the angle values of the single crystals of TPA , TPA(PDAF)3 displayed a larger distortion derived from the steric hindrance. All compounds dissolved well in common solvents such as chloroform, tetrahydrofuran, and dichloromethane. The thermal properties of TPA(PDAF)n  were estimated using thermogravimetric analysis (TGA) and differentialscanning calorimetry (DSC). The DSC curves indicate that three diazafluorene derivatives show no glass phase transition and melting point by heating to 195 °C . The degradation temperatures  of diazafluorene derivatives are 354, 440, and 485 °C, respectively . With increased diazafluorene units, the TPA(PDAF)n show enhanced thermal stability probably to the rigid and steric hindrance structure that suppresses the phase transition process.Td values suggested the good thermal stability of TPA(PDAF)n for the application as charge storage layers in OFET memory devices.The synthesis routes of tertiary alcohols and TPA(PDAF)2.2n  in solutions and films are plotted in Figures S4 and S5 (Supporting Information), with the corresponding data summarized in Table1, TPA(PDAF)2, and TPA(PDAF)3 showed absorption peaks at 308, 311, and 312 nm, respectively  in solutions exhibits emission spectra with peaks at 494, 487, and 483 nm, respectively. Similar with absorption phenomenon, hypsochromic shifts of 28, 18, and 1 nm were observed in film, as Figuren powder under 365 nm UV light. Photoluminescence spectra in several different polar solvents were also measured and shown in Figure S6 (Supporting Information). Three diazafluorene derivatives exhibited obviously solvent‐dependent fluorescence that is the higher polarity of the solvent, the more pronounced red‐shift of the emission maxima, indicating the complicated excited states and energy transfer occur.3 showed the smallest change 3 revealed single molecular behavior creating random molecular distribution, which could be considered as isolated molecular trapping centers, and minor energy transfer occurred between the centers. As a result, it could be beneficial for charge stored and restrain the charge leakage of charges as charge storage layers in OFET memory devices.The optical properties of the compounds were investigated by UV–visible (UV–vis) and photoluminescence (PL) analysis in solution and solid films. The absorption and emission spectra of TPA(PDAF)n n = , 2, 3 inn n = , 2, 3 inn n = , 2, 3 inn films are showed in Figure S7 (Supporting Information), with the electrochemical properties of TPA(PDAF)n  listed in Table 1, TPA(PDAF)2, and TPA(PDAF)3 were −5.40, −5.39, and −5.38 eV, respectively.And the LUMO energy levels were −2.45, −2.59, and −2.69 eV, respectively. As a result, the electrochemical bandgaps were 2.95, 2.80, 2.69 eV, respectively. The energy levels revealed that the introduction of 4,5‐diazafluorene groups could lead to significant reduction in LUMO levels, while less change occurred in HOMO level (only 0.01 eV). Such low LUMO levels could reduce the energy barrier which is defined as the difference value of the LUMO/HOMO levels between semiconductor and charge trapping elements, and facilitate charge injection and avoid the charge escaping from the trapping layer. Besides, in the anodic successive multiple potential scans, as Figure S8 (Supporting Information) shown, TPA(PDAF)3 exhibited most stable reversible oxidation processes. The electrochemical results demonstrated that TPA(PDAF)3 end‐capping with large bulky steric hindrance imparts extraordinarily high electrochemical stability, which is crucial for the stability of electrical devices.The cyclic voltammetry (CV) curves of TPA(PDAF)AF)n n = , 2, 3 li2.3Figure3‐hybridized C atoms, the complete separation of HOMO/LUMO confirms that there are minimal intramolecular interactions between the two moieties. The complete separation of frontier molecular orbitals of diazafluorene derivatives can facilitate the balance of carrier injecting and transporting, and offer ideal hole and electron trapped sites.1, TPA(PDAF)2, TPA(PDAF)3 were also calculated with the values of −0.11, −0.52, −0.68 eV, respectively 3 is about 1.8 nm × 1.6 nm × 0.9 nm based on the single crystal structure . The multidimensional conformation suggests that the end‐capped bulky molecule TPA(PDAF)3 owned highly nonplanar topological configuration, which could suppress π–π stacking and aggregation in the solid states . In this regard, TPA(PDAF)3 may exhibit single nanomolecular behavior with weaker intermolecular interaction, which is critical for charge storage and retention as charge storage layers in OFET memory devices.To further demonstrate the intramolecular charge transport and the influence of carrier transporting groups on the frontier molecular orbitals, all the molecules are evaluated by density functional theory (DFT) calculations. The geometry optimization and frontier molecular orbitals are shown in 2.4Figuren  were used as the charge storage elements. For OFETs, the morphology and surface energy of charge trapping elements are critical factors to influence the growth and crystallinity of overlaying semiconductor. The TPA(PDAF)n  based charge trapping films were fabricated by solution spin‐coating process on bare SiO2 substrates. The atomic force microscopy (AFM) images of the TPA(PDAF)n films  are with water contact angles of 97.19°, 94.88°, and 96.97°, respectively. The hydrophobic surfaces ensure good wetting and well‐formed growth of pentacene on the smooth electret surface.[[qv: 8a]] AFM images of pentacene on top of TPA(PDAF)n films are shown in Figure 1 exhibited a grain domain with the size about 0.2–0.3 µm, while pentacene on TPA(PDAF)2 and TPA(PDAF)3 exhibited a bigger grain domain with the sizes of ≈0.4–0.6 µm and 0.6–1 µm, respectively. It may be ascribed to the weaker intermolecular interaction of TPA(PDAF)3 in solid state that provided a nanoscaled nucleation center for crystallinity of pentacene.n  are shown in Figure S10 (Supporting Information), and the calculated transistor parameters are listed in Table2 V−1 s−1, respectively. The increase of mobility was attributed to the increased crystalline and grain size of pentacene. The transfer curves of these devices showed the initial threshold voltages (Vth) of −4.8, −18.7, −11.2, and ON/OFF current ratios of 5.1 × 104, 11.1 × 105, and 3.96 × 104, respectively, which were enough to distinguish between “0” and “1” digital states clearly.The output and transfer characteristics of the devices based on TPA(PDAF)2.5Vth) is defined as the difference between the Vth of the programming (PGM) and erasing (ERS) states. It should be noted that the thickness of blocking dielectric layer SiO2 is 300 nm, programming voltage would be relatively large. FigureVDS = −30 V) of the memory devices. After applying a negative gate bias (VG = −100 V for 20 ms) under dark conditions, the transfer curves based TPA(PDAF)1, TPA(PDAF)2, and TPA(PDAF)3 were significantly shifted in the negative direction with threshold voltages (Vth) of −55.7, −73.0, and −74.4 V, respectively, serving as the “programming” process. Subsequently, the memory devices were applied reverse gate bias of 50, 50, and 55 V for TPA(PDAF)1, TPA(PDAF)2, and TPA(PDAF)3 based devices with assistance of light illumination for 1 s, respectively. Then the transfer curves were completely shifted to the positive direction and recovered to their initialstate, serving as the “erasing” process. The negative memory windows between PGM and ERS states were calculated to be 50.9, 54.3, and 63.2 V for TPA(PDAF)1, TPA(PDAF)2, and TPA(PDAF)3 based devices, respectively. In the electric‐programming process, when the programming time was changing from 20 ms to 5 s for TPA(PDAF)n  based device, there was no change with the value of ΔVth  provided strong trapping sites, thus displayed a fast trapping speed. Owing to the programming speed limit of our instrument is 20 ms, faster saturation programming speed could be achieved if experimental conditions permitted. The electric‐programming time of TPA(PDAF)n (<20 ms) was suitable compared with other reported charge trapping elements, such as poly(α‐methylstyrene) (PαMS) (<1 µs),[[qv: 3a]] [2‐(9‐(4‐(octyloxy)phenyl)‐9H‐ﬂuoren‐2‐yl)thiophene]3 (WG3)(≈1 s),n  owned the same hole trapping core TPA unit, and similar potential barrier (approximate HOMO levels) for hole carriers transferred, the widest memory windows of TPA(PDAF)3 may be ascribed to the larger torsion angle of triphenylamine unit. As Figure S1 (Supporting Information) showed, the torsion angles of triphenylamine were 111°, 112°, 109° for TPA(PDAF)1, 108°, 109°, 113° for TPA(PDAF)2, and 122°, 113°, 125° for TPA(PDAF)3. The largest torsion angle of TPA(PDAF)3 may create higher barrier for trapping more charges deeply and prevent the recombination of segregated charges in the trapping layer.1, 15.2, 24.5, 38.3, and 54.3 V for TPA(PDAF)2 and 16.1, 28.3, 44.5, and 63.2 V for TPA(PDAF)3, respectively . Namely, the devices exhibited stepwise shifts of transfer curves with the increase of the negative gate bias, which indicated that these memory devices could process the multibit storage function to achieve high storage density in one cell.To investigate the influence from 4,5‐diazafluorene group on transistor memory devices, we applied appropriate gate pulse to lead the shifts of transfer curves, which brought about high conductance (ON) and low conductance (OFF) states. Memory window ΔVth is def1 based device barely shifted in the positive direction. Whereas the TPA(PDAF)2 based device showed slight positive memory windows of 1.0, 2.1, 7.3, and 12.1 V, respectively. Significantly, much bigger memory windows of 8.2, 12.5, 18.3, and 26.2 V were obtained from TPA(PDAF)3, respectively . The shifts of Vth enhanced with the increased proportion of 4,5‐diazafluorene group. TPA(PDAF)3 based device exposited largest positive memory window, because TPA(PDAF)3 has the lowest LUMO energy level due to the strongest electron affinity, thus facilitating more electrons transfer from pentacene to trapping layer. Moreover, TPA(PDAF)3 possesses the maximum electron‐withdrawing 4,5‐diazafluorene group, which helps for more trapped electrons. As a result, considering the ambipolar memory characteristic, under the conditions of −100 V at hole‐trapping mode and 100 V at electron‐trapping mode, about 52, 66, and 89 V total memory windows were obtained from TPA(PDAF)1, TPA(PDAF)2, and TPA(PDAF)3, respectively, shown in Figure n, single‐component charge trapping elements diazafluorene and triphenylamine were also measured and the result electrical characteristics are shown in Figures S13 and S14 (Supporting Information). Although the memory windows can be found in these devices, they were much smaller than the devices with TPA(PDAF)n, which was clearly indicated that the design strategy of small molecular elements was crucial to achieve the memory performance.However, when applying positive gate bias ranging from 40, 60, 80 to 100 V, the transfer curves of TPA(PDAF)Figuren  illustrated in Figure n . According to Fowler–Nordheim (FN) tunneling mechanism, when applying a negative gate electric field (programming process), the holes were induced through pentacene and transferred into trapping layers, which injected into the HOMO of the molecules and trapped by the triphenylamine moiety, as shown in Figure Vth. As is well‐known that photosensitive pentacene can generate excitons by light emission. For OFETs, the photogenerated excitons can be separated by applying appropriate gate voltage to generate holes and electrons. For the erasing process, when the reverse gate bias and light irradiation were applied, the trapped holes were neutralized by the injected photoinduced electrons, resulting in the transfer curve returning to a high conductance state.E0), cationic (E+), and anionic (E−) states of TPA(PDAF)n molecules are summarized in Table S1 (Supporting Information). When the charge trapping molecule combines a positive charge or a negative charge, the cationic state or anionic state of the molecule were obtained. The values of IP and EA were calculated as IP = E0(Q0) – E+(Q+) and EA = E0(Q0) – E+(Q+).[[qv: 25a]] Comparing the values of TPA(PDAF)n, TPA(PDAF)3 has a much lower energy in both charged states , which facilitate the charge injection and the stabilization of trapped charges.[[qv: 25b]] Therefore, TPA(PDAF)3 based device exhibited higher positive and negative memory windows and better charge retention property.The possible mechanism was proposed in Vth) with different programming gate voltage (VProg). All the devices displayed nearly linear fittings with ΔVth as a function of VProg, corresponding to the model of FN tunneling.[[qv: 8b]] As shown in Figure n increased linearly with the enhanced VProg dramatically which suggested TPA(PDAF)n possess strong hole storage capability. For the positive memory windows, the ΔVth of TPA(PDAF)2 and TPA(PDAF)3 increased linearly with the increasing VProg 2. Larger memory windows could be obtained for TPA(PDAF)2 and TPA(PDAF)3 if a higher VProg was applied. However, there is no increase for TPA(PDAF)1 until applying a VProg beyond 100 V. For TPA(PDAF)n trapping elements, the magnitude of positive memory windows could be controlled by the proportion of 4,5‐diazafluorene group.To further analyze the trapping behavior, we plotted the shifts of threshold voltage  according to the equation: Δn = ΔVth·Ci/e, where e is the element charge (1.602 × 10−19 C), Ci is the total capacitance per unit area . Under a programming voltage of VGS = −100 V for 20 ms, the calculated Δn in these memory devices were about 3.69 × 1012 cm−2 for TPA(PDAF)1, 3.91 × 1012 cm−2 for TPA(PDAF)2, and 4.55 × 1012 cm−2 for TPA(PDAF)3, respectively. It suggests that the TPA(PDAF)n based devices have similar hole trapping capacity. The charge trapping density with a magnitude of 1012 in OFET memory devices is better than the nanofloating gate memory devices newly reported.VGS = 100 V with the assist of light for 1 s, the calculated Δn was 7.49 × 1010 cm−2 for TPA(PDAF)1, 5.99 × 1011 cm−2 for TPA(PDAF)2, and 1.29 × 1012 cm−2 for TPA(PDAF)3. The experimental results indicated that the strong electronic deficiency of 4,5‐diazafluorene group could generate more electron trapping sites and high charge trapping density to improve memory performance, while the hole trapping ability depends on the core of the TPA. The memory characteristics of TPA(PDAF)n  based devices are listed in Table n based OFET memory devices have large hole storage windows, tunable electron storage windows, and high charge storage capacity, our approach is a promising strategy for realizing multilevel characteristics memory.We can calculate the charge trapping density  as charge trapping layers were evaluated through write–read–erase–read (WRER) cycles. After applying writing and erasing gate bias, the ON and OFF currents were steady. The ON/OFF current ratios of TPA(PDAF)n  based devices were measured around 102–103 during the WRER examinations, and the devices were tested for over 100 cycles . The retention time is a critical measure of charge storage stability that reflects the duration of the stored charge retained in the charge‐trapping layer. The retention time of the ON and OFF states of the devices at a gate voltage of −35 V was maintained for 104 s with ON/OFF current ratios of 11, 29, and 522, respectively n devices exhibited nonexponential characteristics, which were consistent with the model for long‐term trapped charge decay in most polymer/small molecule elements.] For the erasing process, when the reverse gate bias and light irradiation were applied, the trapped holes were neutralized by the injected photoinduced electrons, resulting in a high conductance state. Due to linking three diazafluorene units which exhibited strong electron affinity, TPA(PDAF)3 possessed the strongest electron trapping ability. The trapped photogenerated electrons in TPA(PDAF)3 were more stable than those in TPA(PDAF)1 and TPA(PDAF)2. So, the erasing state of TPA(PDAF)3 remained relatively stable compared with the TPA(PDAF)1 and TPA(PDAF)2 devices. In consideration of the programmed state, the IDS of TPA(PDAF)3 exhibited better stability than those of TPA(PDAF)1 and TPA(PDAF)2. One possible reason is that in the TPA(PDAF)3 molecule, the TPA core surrounding by three strong electron‐withdrawing 4,5‐diazafluorene groups, which naturally acts as hole‐blocking role, could trap more holes and prevent the stored charge from leaking. The schematic of molecular charge storage mechanism is revealed in Figure 3 element were found. Additionally, owing to three bulky diazafluorene moieties connected to TPA, the larger torsion angle of TPA(PDAF)3 caused high barrier for trapping holes deeply and prevented the leakage of stored holes leading to long‐time charges reserved in the trapping layer.3 film presented single nanomolecular behavior and random molecular distribution, which could be viewed as isolated molecular trapping centers with minor charge delivery. The special behavior was also helpful for charge storage and maintenance. This result reflects the importance of the electronic structure of the charge storage elements on the memory characteristics.The multiple switching stability of the devices using TPA(PDAF)2.6n  into PS for optimized charge trapping layers. As is well‐known, PS is hydrophobic and has a low‐k, which benefits smooth thin‐film charge trapping elements with pretty good mobility. Besides, PS acted as “charge blocking” layers improving the device stability by blocking the charges release, owing to the lower HOMO level of PS (−5.60 eV) and provided a deeper trapping site[[qv: 8b]] that would avoid the charge escaping from the trapping layers.3 blended in PS serving as charge trapping layers were fabricated to optimize a best blending ratio. The transistor and memory characteristics of various wt% TPA(PDAF)3@PS were shown in Table S2 (Supporting Information). With the blending ratio increased, more trapping sites were provided due to the increasing TPA(PDAF)3, but the hole mobility was decreased which reduced the hole injected from pentacene into TPA(PDAF)n.3. The ON/OFF ratio of 10% TPA(PDAF)3@PS device was almost two orders higher than those with other blend ratios devices. Thus, blending ratio of 10% was chosen to optimize the TPA(PDAF)n  elements.In order to obtain better retention property and ON/OFF current ratio, we blend TPA(PDAF)n@PS devices exhibited much higher field‐effect mobilities of 0.20, 0.25, and 0.55 cm2 V−1 s−1, respectively. The drain–source currents of TPA(PDAF)n@PS based OFETs enhanced dramatically, which benefit larger ON/OFF current ratios of 106, 107, and 107, respectively  and Table 1, 25.43 and 33.98 V for TPA(PDAF)2, and 25.46 and 38.30 V for TPA(PDAF)3 were found. Slightly smaller memory windows occurred comparing with isolated TPA(PDAF)n based devices, which were ascribed to the reduced trapping sites in the blend elements. For TPA(PDAF)n@PS based devices, under −100 V programming process, when the programming time was changing from 20 ms to 5 s, the memory windows were also not changed, which revealed the fast trapping efficiency in TPA(PDAF)n 2, and 27.0 V for TPA(PDAF)3 under a positive gate bias of 80 V with assistance of light for 1 s. Whether blend with PS or not, the role of 4,5‐diazafluorene group for electron trapping, high charge trapping density, and preventing hole leakage is still obvious. The ON/OFF current ratios of the three devices were indeclinable along with 100 WRER cycles 1 and TPA(PDAF)2. The ON/OFF current ratio of TPA(PDAF)3 was even up to 1.9 × 104, as shown in Figure 2 and TPA(PDAF)3 based memories exhibited good extrapolated retention characteristics, as Figure 3 based memory still had an ON/OFF current ratio greater than 10.n@PS are promising elements for reliable and stable memory devices used in high density information storage with controllable charge behaviors.As Figure S17 (Supporting Information) showed, the transfer characteristics of the optimized 10 wt % TPA(PDAF)3n  serving as molecular charge storage elements, which possess D–A type conjugate interrupted structures with steric hindrance building blocks. With fully end‐capping steric hindrance of triphenylamine, TPA(PDAF)3 owns highly nonplanar topological configuration and weaker intermolecular interaction, which suppress π–π stacking and aggregation in the solid states. The complete separation of HOMO/LUMO of diazafluorene derivatives facilitates the balance of carrier injecting and transporting and offers ideal hole and electron trapping sites. The OFET memory devices based on solution‐processing TPA(PDAF)n  elements were examined and TPA(PDAF)3 exhibited the best memory performance. With increased diazafluorene units, the larger memory windows were observed, whether for hole storage or electron storage, which illustrated that diazafluorene groups not only provided electron trapping sites, but also acted as hole blocking role for trapping more holes deeply in triphenylamine and prevented charge leakage. TPA(PDAF)3 based device showed an ambipolar memory behavior with a large memory window of 89 V, high hole trapping density and fast trapping speed. Optimizing by mixing with 90% PS, the device exhibited reliable endurance property and good charge retention time. The novel donor–acceptor small molecular elements design of trapping core attached with diazafluorene is a promising candidate for high performance OFET memory.In summary, we have successfully synthesized three diazafluorene derivatives TPA(PDAF)4Chemical and Materials: All the solvents and reagents were purchased from commercial suppliers and used without further purification, unless noted otherwise. All products were purchased by flash column chromatography which was carried out with silica gel (200–300 mesh). Spectrochemical‐grade solvents were used for optical measurements. Bromobenzene and 1,10‐Phenanthroline monohydrate, triphenylamine were obtained from Aldrich Chemical Co. Potassium carbonate, magnesium sulphate, chloroform, and toluene were purchased from Suzhou Rosen Chemical Co., Ltd. without further purification. Dichloromethane was dried by anhydrous sodium under room temperature. Tetrahydrofuran (THF) and toluene were dried over sodium benzophenone ketyl anion radical and distilled under a dry nitrogen atmosphere immediately prior to use. DAFO was synthesized as in our previous work.Mw = 250 000) were purchased from Sigma‐Aldrich and used without further purification. Experimental details including general methods, synthesis, NMR, HRMS, theoretical calculations, and crystallographic data are given in the Supporting Information.Characterization: All compounds were characterized by 1H NMR and 13C NMR on a Bruker AVANCE III 400 MHz instrument with tetramethylsilane as the internal standard. For the high‐resolution mass spectrometry, the data were recorded on Thermo Fisher Scientific LTQ FTICR‐MS with MALDI_DHB positive ion mode. X‐ray diffraction was performed on a Bruker D8 X‐ray diffractometer with Mo Ka radiation (λ = 0.71073 Å), and the operating 2θ angle ranged from 5° to 50°, with the step length of 0.02°. Absorption spectra were measured with a Shimadzu UV‐3150 spectrometer and emission spectra were recorded on a Shimadzu RF‐530XPC luminescence spectrometer. DSC analyses were performed on a Shimadzu DSC‐60A Instrument. TGAs were conducted on a Shimadzu DTG‐60H thermogravimetric analyzer under a heating rate of 10 °C min−1 and a nitrogen flow rate of 20 cm3 min−1. CV studies were conducted using an CHI600C in a typical three‐electrode cell with a platinum sheet working electrode, a platinum wire counter electrode, and a silver/silver nitrate (Ag/Ag+) reference electrode. AFM measurements were obtained with a NanoScope IIIa AFM at room temperature. Commercial silicon cantilevers with typical spring constants of 21–78 N m−1 were used to operate the AFM in tapping mode. All the quantum‐chemical calculations were performed using the Gaussian 09 program suite.Device Fabrication and Characterization: OFET memory devices were fabricated with a top‐contact and bottom‐gate configuration. Heavily doped n‐type Si wafer with 300 nm SiO2 as the gate dielectric (15 nF capacitance) was used as the substrate. The substrates were cleaned sequentially in an ultrasonic bath with acetone, ethanol, and deionized water for 10 min each, and then transferred into an oven at 120 °C for 30 min after dried using a nitrogen gun. Later, the substrate surface was UV/ozone cleaned for 3 min and transferred to a N2 filled glovebox. Chloroform solution of TPA(PDAF)n and TPA(PDAF)n@PS (5 mg mL−1) were stirred 30 min to form a homogeneous solution. All the solution‐processed single‐component films were fabricated by spin‐coating, with the spin speed of 2500 rpm for TPA(PDAF)1, 2900 rpm for TPA(PDAF)2, 3000 rpm for TPA(PDAF)n, and 3000 rpm for TPA(PDAF)n@PS, respectively. The spin time was 30 s. Then, they were baked at 80 °C on a hot plate for 30 min to remove the residual solvent. The thicknesses of TPA(PDAF)n thin films were estimated to be 22.3, 20.8, and 20.3 nm, respectively. The thicknesses of (0–40 wt%) TPA(PDAF)3@PS films were 37.2, 33.6, 33.9, 34.2, 36.7, and 35.0 nm, respectively. The thicknesses of TPA(PDAF)n @PS films were 35.5 and 34.2 nm. After that, 50 nm thick pentacene was thermally evaporated at a deposition rate of 0.1 Å s−1 under a pressure of 5 × 10−4 Pa. Finally, about 50 nm thick Au was thermally evaporated through a shadow mask to form source and drain electrodes with the channel width W = 1500 µm and length L = 100 µm. Film thickness was measured by Bruker Dektak XT stylus profiler. The electrical characteristics of the memory devices were characterized using an Keithley 4200‐SCS semiconductor parameter analyzer. All electrical measurements were carried out under ambient conditions at room temperature.The authors declare no conflict of interest.SupplementaryClick here for additional data file.


Purpose: Tumor cells, with drug resistance, are associated with failed treatment and poor prognosis. Our aim was to explore potential strategy to overcome the epidermal growth factor receptor (EGFR) inhibitors resistance in non-small cell lung cancer (NSCLC).Materials and methods: Flow cytometry was used to examine and sort cells. Using MTT assay, we detected the cell viability under different conditions. Using RT-qPCR and Western blot, we determined the targeted gene expression in mRNA and protein levels. The morphology of the prepared nanoparticles was pictured by transmission electron microscopy. We also performed immunohistochemistry (IHC) and immunofluorescence (IF) to detect the proteins expression. Subcutaneous cancer models in nude mice were constructed to evaluate the anti-cancer effects in vivo.Results: Here, we observed enhanced expression of integrin αvβ3 in tumor tissues from EGFR inhibitors resistant patients. Also, integrin αvβ3-positive NSCLC cells revealed significant EGFR inhibitors resistance, resulting from the activation of Galectin-3/KRAS/RalB/TBK1/NF-κB signaling pathway. Co-encapsulating integrin αvβ3 inhibitor and EGFR inhibitor further improved the drug delivery system, leading to superior anti-cancer effects and reduced systemic toxicity.Conclusion: Our results demonstrated that co-encapsulation of erlotinib and cilengitide by MPEG-PLA (Erlo+Cilen/PP) nanoparticles revealed enhanced tumor suppression along with reduced organ damages, providing an innovative approach for NSCLC treatment. Lung cancer is the most common malignance with a high lethality worldwide , two maiIntegrins, which consist of α subunit and β subunit, are heterodimeric adhesion receptors which are associated with the cellular adhesion to the extracellular matrix and signal transduction. By now, 18 α subunits and 8 β subunits of integrins have been identified in humans, which combine to form 24 different integrin heterodimers . AccumulIn our studies, enhanced expression of integrin αvβ3 was observed in EGFR-resistant NSCLC samples. Additionally, our results revealed that the Galectin-3/KRAS/RalB/TBK1/NF-κB signaling pathway was activated by integrin αvβ3, leading to the EGFR-associated drugs resistance in NSCLC. Concomitantly, the application of integrin αvβ3 inhibitor, cilengitide (Cilen), could efficiently reverse the drug resistance. To further enhance the anti-cancer effects and avoid the side effects, we developed the MPEG-PLA nanoparticles to co-encapsulate Cilen and EGFR inhibitors. Compared with traditional drug delivery system, such as gold nanoparticles  or liposA549 (human NSCLC cell line), NCI-H1975 (human NSCLC cell line) and Lewis (mouse NSCLC cell line) cells were purchased from the Cell Bank of Chinese Academy of Sciences . All cell lines were cultured in RPMI-1640  supplemented with 10% fetal bovine serum , penicillin (100 U/ml) and streptomycin (0.1 mg/ml). Doxorubicin (DOX), paclitaxel (PTX), gemcitabine (GEM), erlotinib (Erlo) ,14, gefiNSCLC samples were obtained by fiberoptic bronchoscopy at The Second Affiliated Hospital of Zhejiang Chinese Medical University and were kept in 4% paraformaldehyde. According to the clinical data, samples were divided into newly diagnosed and EGFR resistance. All samples were confirmed as NSCLC by a pathologist expert. All subjects gave written informed consent. Ethical approval was obtained from the Committee of the Second Affiliated Hospital of Zhejiang Chinese Medical University and all researchers have been carried out in accordance with the World Medical Association Declaration of Helsinki. After fixation, the samples were processed, embedded in paraffin, and sectioned at 4 μm for further study. Antigen retrieval was done using citric acid and sodium citrate in a microwave oven . Then the sections were incubated with integrin αvβ3  at 4°C overnight, followed by signal amplification using an ABC HRP Kit  and counter-staining with Hematoxylin, dehydration with series of graded ethanol and cleaned with xylene. Microscope  was used to visualize the sections. For cell immunofluorescence (IF), samples were blocked in 5% bovine serum albumin in PBS for 1 h, NF-κB  were incubated at 4°C overnight, followed by signal amplification using TSA Kit . An Olympus confocal microscope (Japan) was used to visualize.+ cell sorting, BD FACSARIA III was used to sort PE positive cells.Cells were collected in PBS and stained with anti-human integrin αvβ3 PE  at 4°C for 30 min. To examine the percentage of integrin αvβ3 in patients’ samples, 1 mg/ml collagenase , 2 units/ml hyaluronidase , and 0.1 mg/ml DNase  were used to digest the tissues into single cells, the anti-human CD45 APC  and anti-human integrin αvβ3 PE  were stained at 4°C for 30 min. After washing, the data were collected by BD Canto II . 7-AAD was used to exclude the dead cells. IgG  was used as the negative control. For integrin αvβ3Cell viability was determined by MTT assay. Briefly, 3000 cells were seeded into 96-well culture plates. After 12 h, cells were treated with various concentrations of drugs or nanoparticles. After 48 h, cell growth was measured after addition of 10 μl of 0.5 mg/ml MTT solution. After 4 h of incubation at 37°C, the medium was replaced with 100 μl dimethyl sulfoxide and vortexed for 10 min. Absorbance was measured at 570 nm by a microplate reader (Bio-Rad). Each experiment was performed for at least three times.Erlo+Cilen loaded MPEG-PLA nanoparticles preparation: 95 mg MPEG-PLA, 1 mg Erlo (Lapa or Gefi) and 5 mg Cilen were co-dissolved in 3 ml dichloromethane, followed with evaporation under reduced pressure in a rotary evaporator at 60°C. Then 500 ml PBS (pH = 7.4) was used to rehydrate the film and allow the self-assembly of Erlo+Cilen loaded MPEG-PLA nanoparticles.DL and EE of co-delivery PG NPs were calculated from the following formulas:The particle size of Erlo+Cilen/PP NPs was detected by Malvern Nano ZS90 . The measuring progress was performed under the temperature of 25°C. All results were performed for at least three independent experiments.The morphology of the prepared nanoparticles was pictured by transmission electron microscope . Briefly, the nanoparticles were diluted by distilled water and then placed on a copper grid covered with nitrocellulose. Samples were negatively stained with phosphotungstic acid and dried at room temperature. Each experiment was performed for at least three independent times.In vitro drug release behaviors of free Cilen, free Erlo and Erlo+Cilen loaded nanoparticles were detected by a dialysis method. Briefly, 2.5 ml Cilen or Erlo in PBS, or Erlo+Cilen/PP nanoparticles were placed in dialysis bags . The dialysis bags were incubated in 25 ml PBS containing 2.5 ml fetal calf serum (37°C and pH = 5.0 or 7.4) in a 50-ml centrifugal tube at 100 rpm. At predetermined time points, 2 ml of release medium was collected for the drug release analysis. The amount of released drugs was quantified by high-performance liquid chromatography . For Erlo, an octadecylsilyl silica gel column  was used. The column was maintained at room temperature. The detection wavelength was 316 nm. The mobile phase solvent consisting of acetonitrile, methanol, water and trifluoroacetic acid (26:26:48:0.1) was pumped at a flow rate of 1.0 ml/min. The Cilen release in those samples was determined by HPLC with UV detection , eluent B = acetonitrile/formic acid (999:1), gradient: t = 0 min 90% A, t = 0.6 min 90% A, t = 4 min 10% A, t = 5.5 min 10% A, column temperature 37°C). Each experiment was performed for three independent times.Nude mice  were purchased from Beijing Vital River Laboratory Animal Technology  and received a subcutaneous injection with 1 million A549 cells. One week later, mice were randomly separated into different groups and treated with Erlo  , Gefi  and were boiled for 10 min, then samples were separated by SDS/PAGE, followed by transferring to PVDF membranes and detecting by immunoblotting with primary antibodies against human integrin αvβ3 , Galectin-3 , KRAS , RalB , TBK1 , p-TBK1 , respectively, at 4°C overnight. Then HRP–conjugated secondary antibody  was incubated for 1 h at room temperature, and visualized by using ECL detection kit . β-actin  was used as an internal control.t test by the GraphPad 6.0 software. P-value <0.05 was considered as statistically significant.Results were presented as mean ± SEM and statistical significance was examined by an unpaired Student’s + and αvβ3− cells from NSCLC cell lines  and treated with three clinical EGFR inhibitors . Compared with the αvβ3− or the control groups, the integrin αvβ3+ cancer cells showed significant EGFR inhibitors resistance  effects and selective pH release . MPEG-PLtoxicity G. More itoxicity H,I. The + NSCLC cells, the integrin αvβ3+ A549 cells were inoculated in nude mice to establish the EGFR inhibitors resistant tumor model. Here, we found that Erlo+Cilen/PP efficiently restrained tumor growth and prolonged the tumor-bearing mice survival compared with the free drugs groups  transporters protein, DNA damages repair, expression of anti-apoptotic protein such as BCL-2 expression, and activation of pro-survival pathway induced by cellular surface receptors ,29,30. PIt has been demonstrated that integrins family served as the cancer stem cells markers, which regulate the cancer stemness and promote tumor progressions in a wide range of tumors 31. Our sin vivo. According to previous reports and the optimal proportion of Cilen, Erlo (50 mg/kg) [in vivo. We co-encapsulated the chemotherapeutic agents and integrin αvβ3 inhibitor, and found that the co-encapsulation of EGFR drugs and integrin αvβ3 inhibitor by MPEG-PLA nanoparticles enables the tumor cells to uptake the EGFR drugs and integrin αvβ3 inhibitor simultaneously, which efficiently reverses the drug resistance and enhanced the cytotoxicity to tumor cells. Compared with previous reports, the similar dose of chemotherapeutic agents also induced superior anti-cancer effects after co-encapsulation with αvβ3 inhibitor, along with reduced systemic toxicity. Apart from the advantages of improved drugs delivery systems based on nanoparticles, the MPEG-PLA nanoparticles composed of degradable polyethylene glycol and polylactic acid, which were approved by the U.S. Food and Drug Administration, ensuring the safety as drugs delivery carriers compared with other nanoparticles [Unfortunately, traditional anti-EGFR drugs reveal severe side effects, such as tetter, diarrhea, renal toxicity and so on . To furt0 mg/kg) , Gefi (20 mg/kg) , Lapa (10 mg/kg) , Cilen (articles . MoreoveIn conclusion, we defined the integrin αvβ3 as the driver of EGFR drugs resistance via the activation of RalB/TBK1/NF-κB signaling pathway. We also revealed that co-delivery of integrin αvβ3 inhibitors and EGFR agents by MPEG-PLA nanoparticles could efficiently reverse the drug resistance induced by integrin αvβ3 and provide a promising strategy in the treatment of NSCLC.


Arabidopsis thaliana accession Columbia-0, glucosinolate biosynthesis and turnover are coordinated according to nutrient availability. Whereas exogenous carbon sources had general quantitative effects on glucosinolate accumulation, sulfur or nitrogen limitation resulted in distinct changes in glucosinolate profiles, indicating that these macronutrients provide different regulatory inputs. Raphanusamic acid, a breakdown product that can potentially be formed from all glucosinolate structures appears not to reflect in planta turnover rates, but instead correlates with increased accumulation of endogenous glucosinolates. Thus, raphanusamic acid could represent a metabolic checkpoint that allows glucosinolate-producing plants to measure the flux through the biosynthetic and/or turnover pathways and thereby to dynamically adjust glucosinolate accumulation in response to internal and external signals.Dynamically changing environmental conditions promote a complex regulation of plant metabolism and balanced resource investments to development and defense. Plants of the Brassicales order constitutively allocate carbon, nitrogen, and sulfur to synthesize glucosinolates as their primary defense metabolites. Previous findings support a model in which steady-state levels of glucosinolates in intact tissues are determined by biosynthesis and turnover through a yet uncharacterized turnover pathway. To investigate glucosinolate turnover in the absence of tissue damage, we quantified exogenously applied allyl glucosinolate and endogenous glucosinolates under different nutrient conditions. Our data shows that, in seedlings of  Arabidopsis thaliana, a small herbaceous plant of the Brassicaceae family, is a widely used and well-studied model organism for plant specialized metabolism, particularly the metabolism of glucosinolates  was provided as the only sulfur source, seedlings accumulated higher levels of sulfur-containing metabolites, including glucosinolates, than seedlings on glucosinolate-free, sulfur-deficient medium  is less well understood and the pathways and their regulation are yet to be uncovered  were sterilized with chlorine gas (generated by mixing 100 ml 14% sodium hypochlorite and 3 ml 37% HCL) for 3h. Growth medium was prepared as described in a published protocol (dx.doi.org/10.17504/protocols.io.5q6g5ze). Total nitrogen concentrations were 3 mM (+N) and 0.3 mM (−N) and total sulfur concentrations 1.68 mM (+S) and 0.015 mM (−S). ½ MS medium  as internal standard and subsequently homogenized in a bead mill . After centrifugation , 20 µl of the supernatant was diluted with 180 µL MilliQ-grade water (1:10) and filtered  for the analysis of raphanusamic acid. For glucosinolate analysis, samples were prepared as desulfo-glucosinolates as previously described  coupled to an EVOQ Elite TripleQuad mass spectrometer  equipped with an electrospray ionization source (ESI). The injection volume was 1 µL. Separation was achieved with a gradient of water/0.05% (v/v) formic acid (solvent A)–acetonitrile (solvent B) at a flow rate of 0.4 ml/min at 40°C . The elution profile was: 0–0.5 min 2% B; 0.5–1.2 min 2-30% B; 1.2–2.0 min 30–100% B; 2.0–2.5 min 100% B; 2.5–2.6 min 100–2% B; 2.6–4.0 min 2% B. For raphanusamic acid the instrument parameters were optimized by infusion with the pure standard . The mass spectrometer parameters were as follows: ionspray voltage was maintained at 3,500 V, cone temperature was set to 300°C and heated probe temperature to 400°C, cone gas flow was set to 20 psi, probe gas flow to 40 psi, nebulizer gas 60 psi, and collision gas to 1.5 mTorr. Nitrogen was used as cone and nebulizer gas, and argon as collision gas. Multiple reaction monitoring (MRM) was used to monitor analyte parent ion > product ion transitions [collision energy]: (pos) 164 > 118 [15 V] (quantifier) and 164 > 59 [15 V] . Both Q1 and Q3 quadrupoles were maintained at unit resolution. Glucosinolates were quantified relative pOHb using experimentally determined response factors with commercially available standards in a representative plant matrix. Raphanusamic acid was quantified based on external calibration using a dilution series of an authentic standard.For glucosinolate and raphanusamic acid analysis, seedlings were harvested into 300 µl 85% (v/v) methanol (HPLC grade) containing 10 µM Statistical analyses (parametric and non-parametric standard tests) were performed using the software R  within ten to 13 days after transfer (16 to 19 day-old seedlings) and caused stagnation in glucosinolate accumulation  < 0.001) and nitrogen availability (P(N) < 0.001; In seedlings grown on 50 µM allyl glucosinolate-containing medium, short-chain aliphatic glucosinolates showed the same pronounced increase seven days after germination 8, but aNine days after germination, seedlings grown on 50 µM allyl glucosinolate-containing medium had accumulated >35% higher total levels of glucosinolates than seedlings grown on control medium 8. To coOverall, the presence of sucrose in the growth medium correlates with lowered total glucosinolate levels 17 indepA. thaliana, i.e. at a developmental stage at which biosynthesis and turnover of glucosinolates coincide. Between three and nine days after germination, the average rate of total glucosinolate accumulation was ca. 1 nmol/day . Under nitrogen limiting conditions, the strongest fold-reduction was detected for short-chain aliphatic glucosinolates, which are, however, less abundant than indolic and long-chain aliphatic glucosinolates at this developmental stage  pathway  and Villumfonden, project no. 13169 .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.


FCGR loci. Additionally, we will discuss the role of neutrophils in providing help to marginal zone B cells for the production of antibodies, which in turn may trigger neutrophil effector functions when engaging FcγRs.Neutrophils are best known for their critical role in host defense, for which they utilize multiple innate immune mechanisms, including microbe-associated pattern recognition, phagocytosis, production of reactive oxygen species, and the release of potent proteases, mediators, antimicrobials, and neutrophil extracellular traps. Beyond their well-established contribution to innate immunity, neutrophils were more recently reported to interact with various other cell types, including cells from the adaptive immune system, thereby enabling neutrophils to tune the overall immune response of the host. Neutrophils express different receptors for IgG antibodies (Fcγ receptors), which facilitate the engulfment of IgG-opsonized microbes and trigger cell activation upon cross-linking of several receptors. Indeed, FcγRs (via IgG antibodies) confer neutrophils with a key feature of the adaptive immunity: an antigen-specific cell response. This review summarizes the expression and function of FcγRs on human neutrophils in health and disease and how they are affected by polymorphisms in the  Following a circadian rhythm, neutrophils are released from the bone marrow (Neutrophils are key players of the innate immune response. They are the most abundant leukocytes in the human blood (4.5–11 × 10e marrow –8. This e marrow ] bare thKA) ranging from 8 × 107 down to 2 × 104 M−1  stand out. They bestow on neutrophils the capacity to react in an antigen-specific way—hence to acquire a key feature of the adaptive immunity. FcγRs enable neutrophils to interact with and respond to monomeric or aggregated immunoglobulins, antigen–antibody immune complexes, and opsonized (antibody-coated) particles, cells, or surfaces. Humans express six classical FcγRs: FcγRI/CD64, FcγRIIA/CD32A, FcγRIIB/CD32B, FcγRIIC/CD32C, FcγRIIIA/CD16A, and FcγRIIIB/CD16B . All theFCGR polymorphisms have been described in humans, adding to the complexity of this receptor family with overlapping functions and affinities for their ligands that collaborate, regulate, or compete with each other to tune cellular responses.All FcγRs, except for FcγRIIB and FcγRIIIB, are classical activating receptors. Their activating signals are transduced by an immunoreceptor tyrosine-based activation motif (ITAM) that is either present in the cytoplasmic domain of the FcγR itself (FcγRIIA/FcγRIIC) or in an associated signaling subunit, notably the FcRγ chain. Upon FcγR aggregation by multimeric ligands, Src family kinases phosphorylate these motifs, allowing the activation of a signaling cascade, involving the spleen tyrosine kinase (SYK), phosphatidylinositol 3-kinase (PI3K), phospholipase C (PLC)-γ, Rho, and Rac, resulting in calcium mobilization, cell activation, cytokine/chemokine production, and cell migration –13. CounIn this review, we will focus on IgG receptors (FcγRs) on neutrophils and their role and regulation in steady state and inflammatory conditions.2+ mobilization, cell adhesion, and degranulation, but not to respiratory burst -anchored protein with no signaling capacity on its own. It was first described on neutrophils in 1982 with the means of a newly developed monoclonal antibody  that also recognizes FcγRIIIA on monocytes and NK cells . Incubatry burst –27. The ry burst , 27–29, ry burst –32. In tantibody . The maiantibody . It is gantibody , 35. Desantibody , 37. Dueantibody , the bioantibody . Adding ractions . These iractions .in vitro, neutrophils become activated, degranulate, and produce inflammatory mediators and ROS and trigger neutrophil extracellular trap (NET) formation 2 fragments or ICs can be rather attributed to FcγRIIIA than to the GPI-anchored receptor.In contrast to the abundant FcγRIIIB and FcγRIIA, neutrophils express less than 2,000 copies of the high-affinity FcγRI . Ligatioragments . Many diragments . This reNeutrophil FcγR expression can change dramatically in the context of an inflammation or infection. Notably, FcγRI is strongly upregulated in the presence of inflammatory cytokines such as interferon-γ (IFN-γ) or granulocyte colony-stimulating factor reaching up to 20,000 copies per cell , 54, 55.IFN-γ treatment has little effect on neutrophil FcγRIIA expression but can induce FcγRIIB expression  and, depending on the experimental conditions, may induce FcγRIIIB down-modulation , 71. Fcγin vitro and helped to understand that soluble ICs require primed neutrophils to efficiently trigger external ROS production and degranulation, while insoluble ICs can activate unprimed neutrophils, leading to intracellular ROS production, degranulation, and sustained liberation of inflammatory mediators such as IL-8 and leukotriene B4 (LTB4) that sustain neutrophil-driven inflammation  and copy number variations (CNVs). These genetic variants display heterogeneity among ethnic groups (FCGR2A results in substitution of an arginine residue by a histidine at position 131 (rs1801274) that results in a receptor variant with an improved binding to IgG2 (and to a lesser extent to IgG1 and IgG3) , has been described that retains a cryptic exon in the cytoplasmic tail of the receptor (FCGR2B (rs1050501) results in the replacement of a threonine by an isoleucine in the transmembrane domain (I232T). The presence of threonine in that position entails a failure of FcγRIIB to enter lipid rafts and, as a consequence, reduces its ability to inhibit activatory receptors  located in the membrane-distal Ig-like domain. These variants are termed NA1 (R36 N65 A78 D82 V106), NA2 (S36 S65 A78 N82 I106), and SH (S36 S65 D78 N82 I106)  . They do82 I106)  and is a82 I106) . The SH  numbers  that cou numbers .FCGR2/3 locus  of the resulting receptor . Indeed, a high degree of linkage disequilibrium has been reported for the /3 locus , 98, 99 /3 locus . Associa/3 locus . Adding eceptors . These Creceptor . CNVs ofimmunity –106. Indtibility , 108, whtibility , 110.FCGR3B (and the FCGR2C gene)  , 107. Whwith SLE , 112. Thwith SLE . This suThe best-described pathway of inhibiting activating FcγRs is, without doubt, through co-engagement of the inhibitory FcγRIIB by the same immune complexes , 115. Hoin vivo  or anti-EGFR mAb (cetuximab). Notably, copy numbers of FcγRIIIB could be linked to the inhibitory effect and blocking FcγRIIIB with F(ab′)2 3G8 improved target cell killing , and sialylated IgG has reduced capacity to initiate ADCC in vivo  , a molectibodies .BH) are located around the MZ of healthy human donors and express high levels of the B cell-stimulating cytokines BAFF, APRIL, and IL-21 in response to microbial stimuli and thereby provide help to MZ B cells to trigger antibody production against T cell-independent antigens  and the red pulp –170. Theantigens . In patisubjects . Contradt donors ; also, st donors . In addit donors . PTX3 ist donors . PTX3 bit donors . The capCollectively, the discussed literature exemplifies how our understanding of neutrophils has evolved from their early descriptions as simple first-line defense cells, equipped with powerful weapons to defend the host but unable to differentiate between different threats, to portrayals of cells capable of tailoring their responses according to their environment. We now appreciate that neutrophils interact with various other cell types, integrate complex stimuli, and cooperate with other players of the immune system to fine-tune their responses. The role and regulation of FcγR on neutrophils is not an exception to this rule. Early reports frequently suggested that IgG ICs had very strong neutrophil-activating capacities, but more recently, a much more detailed picture has been drawn, taking into account the size, solubility, composition, and location of ICs. Furthermore, IgG enables neutrophils to function as antigen-specific cells; at the same time, accumulating evidence suggest that neutrophils in turn tune the activity of (at least some) B cells to regulate antibody production. Much remains to be discovered and hopefully new techniques will allow us to unravel previously unappreciated functions of neutrophils and revise others. We are looking forward to hearing more about these exciting cells.All authors listed have made a substantial, direct and intellectual contribution to the work, and approved it for publication.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.


Background: Nimodipine can block the influx of calcium into the vascular smooth muscle cell and prevent secondary ischemia in patients with aneurysmal subarachnoid hemorrhage. However, the reduction of blood pressure after long-term intravenous administration of nimodipine has been associated with neurological deterioration. Yet, no effective solutions have been suggested to address this phenomenon. The use of neuroprotective drug combinations may reduce the risk of sudden blood pressure loss. This prospective, randomized, controlled trial was performed to evaluate the nimodipine-sparing effect of perioperative dexmedetomidine infusion during aneurysmal subarachnoid hemorrhage.Methods: One hundred nine patients who underwent aneurysm embolization were divided into three groups: group C , group D1 , and group D2 . Patient-controlled analgesia was given for 48 h after surgery. The primary outcome measure was the total consumption of nimodipine during the first 48 h after surgery. The secondary outcome measures were recovery time at post-anesthesia care unit (PACU), postoperative pain intensity scores, dexmedetomidine and sufentanil consumption, hemodynamic, satisfaction of patients and neurosurgeon, neurologic examination , Bruggemann comfort scale, and adverse effects. Intraoperative hemodynamics were recorded at the following time-points: arrival at the operating room (T1); before intubation (T2); intubation (T3); 5 min (T4), 10 min (T5), and 15 min (T6) after intubation; suturing of femoral artery (T7); end of surgery (T8); extubation (T9); and 5 min (T10), 10 min (T11), and 15 min (T12) after arrival at the PACU. The level of sedation was recorded at 15 min, 30 min, 1 h, and 2 h after extubation. We also recorded the incidence of symptomatic cerebral vasospasm during 7 days after surgery, Glasgow Outcome Score (GOS) at 3 months, and incidence of cerebral infarction 30 days after surgery.Results: The consumption of nimodipine during the first 48 h after surgery was significantly lower in group D2 (P < 0.05). Compared with group C, HR and MAP were significantly decreased from T2 to T12 in group D1 and D2 (P < 0.05). Patients in group D2 showed a significantly decreased MAP from T5 to T9 compared with group D1 (P < 0.05). The consumption of sevoflurane, remifentanil, dexmedetomidine, and nimodipine were all significantly reduced in groups D1 and D2 during surgery (P < 0.05). Compared with group C, MAP was significantly decreased in groups D1 and D2 during the first 48 h after surgery (P < 0.05). Compared with group C, consumption of sufentanil and dexmedetomidine at 1 h, pain intensity at 1 h, and 8 h after surgery were significantly decreased in groups D1 and D2 (P < 0.05). FAS was significantly higher in group D2 at 8 h, 16 h, and 24 h after surgery. LOS was significantly lower only in group D2 at 0.5 h after surgery (P < 0.05). Compared with group C, BCS was significantly higher group D2 at 4 h and 8 h after surgery (P < 0.05). There were no significant differences among the three groups in consumption of propofol, cisatracurium, fentanyl, and vasoactive drugs during operation, recovery time at PACU, satisfaction of patients and neurosurgeon, and number of applied urapidil and GCS during the first 48 h after surgery. The incidence of symptomatic cerebral vasospasm during 7 days after surgery, GOS of 3 months, and cerebral infarction after 30 days were also comparable among the three groups.Conclusions: Dexmedetomidine  significantly reduced the total consumption of nimodipine during the first 48 h after surgery and promoted early rehabilitation of patients although the incidences of symptomatic cerebral vasospasm, GOS, and cerebral infarction were not reduced. Aneurysmal subarachnoid hemorrhage (aSAH) is an acute cerebrovascular condition associated with high morbidity and mortality worldwide . It impovia enteral administration except in cases of impaired absorption or metabolism (Previous studies have strongly suggested that nimodipine should be administered tabolism . Howevertabolism . Accorditabolism .Recent studies reported that dexmedetomidine may have a neuroprotective effect and is safe for neurosurgery such as craniotomy and carotid endarterectomy (CEA) . The genFollowing our search for English language articles published between 1980 and 2017 on MEDLINE, PubMed, Embase, and Cochrane Central Register of Controlled Trials using the terms of dexmedetomidine, nimodipine, aneurysmal subarachnoid hemorrhage, and cerebral vasospasm, we did not find any study on the nimodipine-sparing effect of perioperative dexmedetomidine infusion during aneurysmal subarachnoid hemorrhage. Therefore, we conducted this prospective, randomized, controlled trial on this topic.We obtained ethical approval from the Institutional Review Board of Liaocheng People’s Hospital to conduct this prospective, randomized, controlled trial. Written informed consent was obtained from patients or their guardian before participation in this trial. The trial was also registered at chictr.org . All methods were performed in accordance with the relevant guidelines.2), and operative time <1 h or >3 h.Patients admitted to our hospital within 72 h of initial subarachnoid hemorrhage from October 2016 to June 2018 were included in this trial if they met the following criteria: age 40–75 years, American Society of Anesthesiologists (ASA) grades I to III, diagnosed with aSAH using digital subtraction angiography (DSA), undergoing aneurysm embolization with general anesthesia, using patient-controlled analgesia (PCA) for at least 48 h after surgery. Patients who met the following criteria were excluded: reoperation within 48 h, taking drugs known to interact with nimodipine such as anticonvulsants within the last month before enrollment, serious systemic diseases of liver and kidney, psychiatric disorders, ischemic heart disease or second- or third-degree atrioventricular block, long-term abuse of alcohol, opioids, or sedative-hypnotic drugs, obesity , group D2 , and group C . A team of nurses who offered the Acute Pain Services were blinded to this trial, and they prepared the experimental drug (dexmedetomidine was diluted to 50 ml using 0.9% sodium chloride in groups D1 and D2) and the PCA pump (sufentanil (0.01 μg·kg–1·h–1) plus dexmedetomidine (0.1 μg·kg–1·h–1) programmed to deliver a bolus of 2 ml, with a background infusion of 2 ml·h–1, a lockout of 5 min, and 1-h limit of 16 ml). All anesthesiologists and neurosurgeons who participated in this trial were blinded.A computer-generated randomization table was used to allocate patients into three groups by an independent anesthetist before surgery: group D1  was infused for 10 min in three groups, fentanil (2–3 μg·kg–1), propofol (1.5–2 mg·kg–1), and cisatracurium (0.2 mg·kg–1) were administered intravenously, and tracheal intubation was performed 3 min later. Sevoflurane (1.5–2.1%), remifentanil (0.05–0.1 μg·kg–1·min–1), and nimodipine (5–20 μg·kg–1·h–1) were administered during surgery. Cisatracurium (0.05 mg·kg–1) was intermittently added to maintain muscle relaxation. The pressure of arterial carbon dioxide (PaCO2) was maintained at 35–40 mmHg during surgery. Sevoflurane and the experimental drugs were stopped when all of the coils were placed. Remifentanil was continued until the femoral artery was sutured. All patients received 5 mg of tropisetron and underwent routine reversal of neuromuscular blockade (atropine 4 μg·kg–1 plus neostigmine 15 μg·kg–1) at the end of surgery.All patients had not been premedicated. Peripheral venous was established in the left upper extremity before the start of anesthesia, then five-lead electrocardiogram, noninvasive blood pressure, oxygen saturation, temperature, and bispectral index  were continuously monitored using an automated system . The anesthetic methods adopted are routinely used in our center . Oxygen –1·min–1 stepwise titration according to acceptable hemodynamic limits. Nimodipine was adjusted by 5-μg·kg–1·h–1 stepwise titration if premise of satisfactory depth of anesthesia and poor response to remifentanil. Besides, phenylephrine, ephedrine, atropine, and urapidil were used where necessary. Endovascular embolization was performed by the same neurosurgeon who had more than 15 years of residency experience.The concentration of sevoflurane was adjusted by 0.2% stepwise titration according to BIS. Remifentanil infusion was adjusted by 0.01-μg·kgAll patients underwent head computed tomography (CT) scan immediately after surgery to detect acute complications such as hemorrhage before they were transferred to the PACU. The same neurosurgeon performed neurological examination and trained the patients to operate the PCA pump. Patients were encouraged to push the bolus of PCA when VASm >3. For patients with poor response to PCA, 1 mg butorphanol was administered intravenously.–1·h–1 to maintain hemodynamic parameters after surgery according to the standards of “Guidelines for the Management of Aneurysmal Subarachnoid Hemorrhage: A Guideline for Healthcare Professionals from the American Heart Association/American Stroke Association” , consumption of dexmedetomidine and sufentanil, hemodynamic satisfaction of patients and neurosurgeon, neurological examination , Bruggemann comfort scale  and adverse effects.Intraoperative hemodynamic data (MAP and HR) were recorded at the following time-points: arrival at the operating room (T1); before intubation (T2); intubation (T3); 5 min (T4), 10 min (T5), and 15 min (T6) after intubation; suturing of femoral artery (T7); end of surgery (T8); extubation (T9); and 5 min (T10), 10 min (T11), 15 min (T12) after arrival at the PACU. Level of sedation  was recorded at 15 min and 30 min and 1 h and 2 h after extubation. We also recorded the incidence of symptomatic cerebral vasospasm during 7 days after surgery, Glasgow Outcome Score (GOS) at 3 months, and incidence of cerebral infarction 30 days after surgery.According to our previous study, the sample size was calculated on the basis of an expected difference of 20% in the cumulative amount of nimodipine utilized 48 h after surgery. For a study power of 80% , the required sample size per group was calculated to be 32 . Assuming a dropout rate of 10%, the final sample size was determined to be 35 patients for each group.post hoc multiple comparisons. The nonparametric Kruskal–Wallis test was used for non-normally distributed continuous variables presented as an inter-quartile range. Categorical data were expressed as frequencies and percentages and analyzed using chi-squared tests or Fisher’s exact tests. P < 0.05 was considered statistically significant.Statistical analyses were carried out using SPSS for Windows Version 21.0 . The Kolmogorov–Smirnov test was used to assess the distribution of the variables. Homogeneity of variance was determined using Levene’s test. Normally distributed continuous variables were presented as mean ± SD. Inter-group comparisons were performed using repeated-measures analysis of variance. The Bonferroni’s correction was used for 2; and six patients had an operative time >3 h. Finally, 120 patients were included in the primary analysis, among whom 11 were excluded . At last, 109 remaining patients were divided into three groups: 35 patients for group C, 38 patients for group D1, and 36 patients for group D2. Three groups were comparable in terms of age, BMI, ASA grade, sex, comorbidity, and GCS before surgery (P > 0.05) . Compared with group C, patients in groups D1 and D2 showed significantly decreased HR and MAP from T2 to T12 (P < 0.05). Patients in group D2 showed a significant decrease in MAP from T5 to T9 compared with group D1 (P < 0.05)  Figure 2vs. 1.50 [1.50–1.70] vs. 1.50 [1.50–1.50] %, P = 0.007), remifentanil , dexmedetomidine , and nimodipine  were significantly reduced in groups D1 and D2—these differences were also statistically significant between groups D1 and D2 (P > 0.05)  (P < 0.05), while MAP was significantly decreased in groups D1 and D2 during the first 48 h after surgery (P < 0.05) (P = 0.021), 16 h (P = 0.007), and 24 h (P = 0.001) after surgery (The consumption of nimodipine during the first 48 h after surgery was significantly lower in group D2 (63.36 [60.48–69.12] < 0.001) Table 4.P < 0.05) (P < 0.05) (P < 0.05) (P < 0.05) (P > 0.05)  Figure 4–1 for 10 min, then adjusted to 0.4 μg·kg–1·h–1 during the surgery) significantly reduced the total consumption of nimodipine during the first 48 h after surgery without any related complications. We also found that the requirements of sevoflurane, remifentanil, and nimodipine in group D2 were significantly decreased during the operation. Although more patients in group C needed vasoactive drugs during the first 48 h after surgery, there was no significant difference among the three groups. Compared with group C, FAS at 8–24 h, BCS at 4–8 h, pain intensity at 1 h, and 8 h after surgery were significantly improved in group D2. LOS in group D2 was significantly lower at 0.5 h after surgery, which may be attributed to the lower consumption of sufentanil and dexmedetomidine. We found no significant differences among the three groups in recovery time at PACU, the satisfaction of patients and surgeon, GCS, the incidence of symptomatic cerebral vasospasm during 7 days after surgery, GOS at 3 months, and cerebral infarction after 30 days.We report, for the first time, that high dose of dexmedetomidine  seemed to be outweighed by the hemodynamic effect. Hence, the author proposed that administration of a plasma expanding drug could be an appropriate strategy to reduce the risk of sudden initial BP reactions  was associated with neurological worsening after intravenous administration of 2 mg·heactions . We alsoeactions . Only oneactions . Dexmedeeactions . HoweverSince 1991, an increasing number of patients choose intravascular embolization to treat aSAH. Compared with surgical clipping, both short- and long-term morbidity and mortality rates are reduced in patients treated with endovascular coiling . We recrAdditionally, McNeill showed that the 6-month mortality of SAH is inversely related to the treatment institution. High-volume hospitals (>60 cases per year) improve the outcomes of patients by provision of neurocritical care units . In our There are several limitations in this trial. First, the number of patients included in this trial is small and hence these results need to be confirmed by larger multicenter randomized controlled trials. Second, we did not measure the serum concentration of dexmedetomidine in this trial as a result of high hospital costs. Third, this trial only studied one dosage of dexmedetomidine-sufentanil after neurosurgery; different dosages should be further investigated. Finally, further studies on specific clinical monitoring for SAH patients are needed, especially for poor-grade patients.–1 for 10 min, then adjusted to 0.4 μg·kg–1·h–1 during surgery) significantly reduced the total consumption of nimodipine during the first 48 h after surgery and promoted early rehabilitation of patients although the incidence of symptomatic cerebral vasospasm, GOS, and cerebral infarction were not reduced. However, more multi-center prospective studies are required to determine the optimal dosage of dexmedetomidine during the perioperative period of neurosurgery.In summary, we report for the first time, that high dose of dexmedetomidine .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.


Enterobacteriaceae, is likely to be used during renal replacement therapy. The aim of this work was to describe the extracorporeal removal (adsorption and clearance) of meropenem-vaborbactam during continuous venovenous hemofiltration (CVVH).The combination product meropenem-vaborbactam, with activity against KPC-producing carbapenem-resistant  Enterobacteriaceae, is likely to be used during renal replacement therapy. The aim of this work was to describe the extracorporeal removal (adsorption and clearance) of meropenem-vaborbactam during continuous venovenous hemofiltration (CVVH). An ex vivo model was used to examine the effects of a matrix of operational settings. Vaborbactam did not adsorb to AN69  ST100  hemofilter; the mean (±standard deviation [SD]) meropenem adsorption was 9% (±1%). The sieving coefficients (mean ± SD) with AN69 ST100 and ST150  filters ranged from 0.97 ± 0.16 to 1.14 ± 0.12 and from 1.13 ± 0.01 to 1.53 ± 0.28, respectively, for meropenem and from 0.64 ± 0.39 to 0.90 ± 0.14 and 0.78 ± 0.18 to 1.04 ± 0.28, respectively, for vaborbactam. At identical settings, vaborbactam sieving coefficients were 25% to 30% lower than for meropenem. Points of dilution, blood flow rates, or effluent flow rates did not affect sieving coefficients for either drug. However, doubling the effluent flow rate resulted in >50 to 100% increases in filter clearance for both drugs. Postfilter dilution resulted in 40 to 80% increases in filter clearance at a high effluent flow rate , compared with ∼15% increases at a low effluent flow rate  for both drugs. For all combinations of setting and filters tested, vaborbactam clearance was lower than that of meropenem by ∼20 to 40%. Overall, meropenem-vaborbactam is efficiently cleared in CVVH mode.The combination product meropenem-vaborbactam, with activity against KPC-producing carbapenem-resistant  Enterobacteriaceae (CRE)  priority list for research and development of novel antibiotics . One proae (CRE) . In Auguae (CRE) , followiae (CRE) . This meefficacy . Compareefficacy .Patients with KPC-producing CRE infections may develop severe renal impairment that necessitates the use of continuous renal replacement therapy (CRRT) . Thus, t14–The aims of this work were (i) to estimate the extent of adsorption of meropenem and vaborbactam within a clinically used continuous venovenous hemofiltration (CVVH) circuit system and (ii) to describe the effect of point of dilution and a range of common CVVH settings on the extracorporeal removal of meropenem and vaborbactam.ex vivo model of CVVH with a clinically used CRRT machine and human blood-crystalloid mixture to closely simulate the clinical condition. The ex vivo model was set up with the commonly used AN69  hemofilters, ST100  or ST150 .Given the difficulty of conducting a clinical study in which the various operational settings of CVVH are altered in a controlled manner so as to describe direct effects on adsorption and/or clearance of drugs, we used an Initial “plasma” concentrations (mean ± standard deviation [SD]) measured from blood-crystalloid mixtures prepared at 50 mg/liter were 53.4 ± 3.5 mg/liter and 60.2 ± 7.3 mg/liter for meropenem and vaborbactam, respectively. These plasma concentrations, which do not affect result analysis, are expected given that the hematocrit of the blood-crystalloid mixture was approximately 20% and the possibility of red blood cell partitioning. For a blood-crystalloid mixture spiked to 50 mg/liter, circulating around a closed ST100 circuit over 3 h, the meropenem concentration decreased relative to that in the stability control experiment . The perThe effect of point of dilution on meropenem and vaborbactam CVVH clearance (with ST100 filter) for different blood flow rates and effluent flow rates is illustrated in in vivo pharmacokinetic properties with no interactions that affect each other's disposition, an important feature suitable for fixed-dose combination  was the same as a previous estimate of 9% . For both filters, a sieving coefficient of ∼1.0 or more shows that meropenem is extensively filtered during CVVH. Vaborbactam filtration was also efficient, although the sieving coefficients were consistently lower than those of meropenem for all filters and conditions tested (mostly about 0.8  ×100.The percent drug remaining after loss by adsorption and degradation was estimated from initial and measured concentrations during the adsorption experiment by using the following equation: percent drug remaining =  ×100.The percent drug adsorbed at each time point was calculated by the following equation using mean percent drug lost by degradation and mean percent drug remaining: percent adsorbed = 100 − percent drug remaining − percent drug lost by degradation.ex vivo experimental model of CVVH was set up with a clinically used Prismaflex CRRT system . Unlike in the adsorption experiment, the ultrafiltrate was directed into the effluent bag and replacement fluid was introduced into the circuit by either predilution or postdilution. Priming fluid and blood-crystalloid mixture were prepared as described above. The Prismaflex with an AN69 filter set was primed in CVVH mode with the following settings: blood flow, 200 ml/min; pre-blood pump, 0 ml/min (no citrate anticoagulation); dialysis fluid, 0 ml/min ; replacement fluid, 1,000 ml/min; percent predilution, 100%; and patient fluid removal, 0 ml/h (hence effluent flow rate matches the replacement fluid rate). Drug-spiked blood-crystalloid mixture was subsequently introduced with priming solution directed to waste, as described above, and the circuit was allowed to equilibrate for 20 min. Clearance experiments were run for different filters and settings in three replicates. Using an AN69 ST100 filter  and initial meropenem and vaborbactam concentrations of either 50 and 50, 20 and 20, or 5 and 5 mg/liter, clearance experiments were run with a 2 × 3 CVVH setting matrix, including blood flow rates of 200 and 100 ml/min against replacement fluid (effluent) flow rates of 1,000, 2,000, and 4,000 ml/h. Similarly, using an AN69 ST150 filter  and meropenem and vaborbactam concentrations of 50 and 50 mg/liter, clearance was assessed for a 1 × 3 CVVH setting matrix of blood flow rate (200 ml/min) against replacement fluid (effluent) flow rates . These experimental conditions were tested with both prefilter and postfilter introduction of replacement fluid.An For each specific setting, the circuit was allowed to equilibrate for 10 min prior to sampling. Samples were taken from the mixing chamber (1 ml of blood), the prefilter sampling port (1 ml of blood), the postfilter sampling port (1 ml of blood), and the effluent sampling port (1 ml of effluent). Blood samples were centrifuged  and the plasma was stored frozen (−80°C) within 15 min. Effluent samples were transferred directly to labeled cryovials and frozen (−80°C) within 15 min. Samples were kept frozen until drug assay.Extracorporeal clearance was calculated as follows: extracorporeal clearance = (effluent drug concentration/mixing chamber drug concentration) × effluent flow rate. The sieving coefficient was calculated as follows: sieving coefficient = effluent drug concentration/[(prefilter plasma concentration + postfilter plasma concentration)/2].Total concentrations of meropenem and vaborbactam in plasma and renal replacement fluid effluent (RRTE) were measured by a validated high-performance liquid chromatography (HPLC)-tandem mass spectrometry (MS/MS) method on a Shimadzu Nexera2 ultrahigh-performance liquid chromatography (UHPLC) system coupled to a Shimadzu 8030+ triple quadrupole mass spectrometer. Each sample (plasma [10 μl] or RRTE [10 μl]) was spiked with internal standard (sulbactam and d6-meropenem) and proteins were precipitated with acetonitrile. An aliquot of 0.2 μl of the supernatant was injected onto the HPLC-MS/MS. The stationary phase was a Poroshell HPH-C18 2.1- by 50-mm, 2.7-μm column  operated at room temperature. The mobile phase A was 0.1% (vol/vol) formic acid in water, and mobile phase B was 100% acetonitrile with 0.1% (vol/vol) formic acid. The mobile phase was delivered with gradient from 5% to 50% at a flow rate of 0.4 ml/min for a run time of 5.5 min. Meropenem was monitored by positive mode electrospray at multiple reaction monitoring (MRMs) of 384.0 → 68.10 (384.0 → 141.10 as reference ion). Deuterated meropenem (d6-meropenem) was monitored in positive mode at 390.15 → 67.90 (390.15 → 147.00 as reference ion). Vaborbactam and sulbactam were monitored by negative mode electrospray at MRMs of 296.25 → 234.2 (296.25 → 278.15 as reference ion) and 232.35 →139.90, respectively. The method was validated for the calibration range from 0.05 to 100 mg/liter. Precision and accuracy of meropenem and vaborbactam were within the industry-standard limits of 15% at all four QC concentrations monitored (