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Encouraging advances in both regenerative medicine and tissue engineering with stem cells require a short-term preservation protocol to provide enough time for quality control or the transportation of cell products from manufacturing facilities to clinical destinations. The hypothermic preservation of stem cells under refrigerated conditions (2-8 °C) in their specific culture medium provides an alternative and low-cost method for cryopreservation or commercial preservation fluid for short-term storage. However, most stem cells are vulnerable to hypothermia, which might result in cell damage from the cooling process and the lack of extracellular matrix (ECM). Herein, we report a peptide scaffold cell-culture-medium additive for mimicking

Human platelets (PLTs) are vulnerable to unfavorable conditions, and their adequate supply is limited by strict transportation conditions. We report here that PLTs preserved under three-dimensional (3D) conditions using novel biomimetic nanofiber peptides showed reduced apoptosis compared with classical PLTs stored at 22 °C and facilitated the storage and transportation of PLTs. The mechanism of PLT 3D preservation involves the formation of cross-links and a 3D nanofibrous network by a self-assembled peptide scaffold material at physiological conditions after initiation by triggers in plasma. PLTs adhere to the surface of the nanofibrous network to facilitate the 3D distribution of PLTs. The 3D microstructure, rheological properties, and effect on the inflammatory response and hemolysis were evaluated. Compared to traditional PLTs stored at 22 °C, PLTs subjected to 3D preservation showed similar morphology, number, aggregation activity, and reduced apoptosis. The detection of the reactive oxygen species (ROS) levels demonstrated that both reduced intracellular and mitochondrial ROS levels were correlated with reduced apoptosis. This study reveals a new 3D preservation method for PLTs based on the use of novel biomimetic nanofiber peptides that presents an attractive opportunity for various biomedical applications.

The sulfur-coordinated organoiridium(III) complexes pbtIrSS and ppyIrSS, which contain C,N and S,S (dithione) chelating ligands, were found to inhibit breast cancer tumorigenesis and metastasis by targeting Wnt/β-catenin signaling for the first time. Treatment with pbtIrSS and ppyIrSS induces the degradation of LRP6, thereby decreasing the protein levels of DVL2, β-catenin and activated β-catenin, resulting in downregulation of Wnt target genes CD44 and survivin. Additionally, pbtIrSS and ppyIrSS can suppress cell migration and invasion of breast cancer cells. Furthermore, both complexes show the ability to inhibit sphere formation and mediate the stemness properties of breast cancer cells. Importantly, pbtIrSS exerts potent anti-tumor and anti-metastasis effects in mouse xenograft models through the blockage of Wnt/β-catenin signaling. Taken together, our results indicate that pbtIrSS has great potential to be developed as a breast cancer therapeutic agent with a novel mechanism.

Fabricating dye adsorbents with efficient adsorption properties is of great significance in the treatment of printing and dyeing wastewater. Herein, composite materials of polydopamine decorated cellulose fibrous nonwovens (PDA@CF NWs) were fabricated by constructing a PDA functional layer on the surface of cellulose fibers

Abstract: The previous year's worldview for cancer treatment has advanced from general to more precise therapeutics approaches. Chemotherapies were first distinguished as the most reliable and brief therapy with promising outcomes in cancer patients. However, patients could also suffer from severe toxicities resulting from chemotherapeutic drug usage. An improved comprehension of cancer pathogenesis has led to new treatment choices, including tumor-targeted therapy and immunotherapy. Subsequently, cancer immunotherapy and targeted therapy give more hope to patients since their combination has tremendous therapeutic efficacy. The immune system responses are also initiated and modulated by targeted therapies and cytotoxic agents, which create the principal basis that when targeted therapies are combined with immunotherapy, the clinical outcomes are of excellent efficacy, as presented in this review. This review focuses on how immunotherapy and targeted therapy are applicable in cancer management and treatment. Also, it depicts promising therapeutic results with more extensive immunotherapy applications with targeted therapy. Further elaborate that immune system responses are also initiated and modulated by targeted therapies and cytotoxic agents, which create the principal basis that this combination therapy with immunotherapy can be of great outcome clinically.

Abstract: There has been a great amount of advancement in the early field of nano-immunotherapy and combination therapy. Persistent consideration regarding the clinical challenges and therapeutic hindrance should be tended to achieve therapeutic efficacy and potential. In this review, we will ad-dress how nanotechnology could defeat the difficulties resulting from cancer immunotherapy, how nanoparticles’ utilization can enhance the efficacy of immune checkpoint blockers, and reconstituting the tumor microenvironment can promote antitumor responses. Moreover, this review discusses how nanoparticles mediate therapeutic modalities like chemotherapy, photodynamic therapy, photother-mal therapy, and radiotherapy, which are used to target and destroy cancerous cells, initiate the release of tumor antigens, and can trigger anti-tumor immunity reactions. Furthermore, we analyzed the po-tential benefits of immunotherapy combinatorial using the nanoparticle delivery system to prevent tumor recurrence, hinder metastases, and decrease systemic toxicity of major organs and healthy cells common with uncontrolled targeting.

Hydrophobic interaction chromatography (HIC) as an indispensable method for protein purification has attracted considerable attentions of researchers as well as biopharmaceutical industries. However, the operation pressure-drop and flow rate of the typical HIC porous resin beads are less than satisfactory. To address these issues, composite nanofibrous membranes (CNFMs) with controllable wettability have been developed by multi-component electrospinning of polystyrene/polyacrylonitrile (PS/PAN), and further by regulating the solution properties and introducing SiO 2 nanoparticles in, CNFMs with proper wettability and large surface area of 23.6 m−2 g−1 were fabricated. The obtained CNFMs showed a high static adsorption capacity of 78 mg g−1 and fast adsorption process within 30 min. Additionally, a high saturation dynamic binding capacity of 39.6 mg ml−1 was obtained under a low pressure-drop of 200 Pa, which surpassed the commercial HIC membrane media. Moreover, these membranes have a selectivity toward the protein with higher hydrophobicity , and proteins adsorbed on the membranes can be eluted with 10 wt% ethanol aqueous solution. Therefore, such a successful fabrication of nanofibrous membranes with outstanding hydrophobic protein adsorption performance could provide a promising platform to prepare economical and inspiring HIC media in the future.

Background The pathogenesis of transfusion-related acute lung injury (TRALI) progress is incompletely understood, and specific therapies for TRALI are lacking. Alveolar macrophages (AMs) are critical for initiation and resolution of lung inflammation. However, the role of AMs in the pathogenesis of TRALI-associated lung failure is poorly understood. Study design and methods Mouse model for in vivo imaging of interleukin (IL)-6 activation in AMs was established by intratracheal instillation of a lentiviral vector carrying the luciferase reporter gene. The TRALI mouse model was produced by intraperitoneal lipopolysaccharide plus intravenous major histocompatibility complex Class I monoclonal antibody treatment. We focused on the changes in AMs in the lung during TRALI and examined whether targeting AMs is an effective strategy to alleviate this condition. Measurements and main results We confirmed that TRALI progress is accompanied by IL-6 activation in AMs. Further study showed that AMs undergo M1 activation during TRALI progress. AM depletion protected mice from TRALI, and transfusion of M1-polarized AMs into 34-1-2 s-treated mice elevated acute lung injury, indicating that the severity of TRALI was able to be ameliorated by targeting AM polarization. Next, we showed that α1 -antitrypsin (AAT) expression improved lung injury by modulating the production of IL-6 in AMs and decreased polarization of AMs toward the M1 phenotype. Conclusions M1-polarized AMs are crucial in a mouse model of TRALI, and AAT may serve as a future treatment for TRALI by regulating the polarization of AMs.

The development of chromatographic media with superb adsorption capacity and large processing throughput is of great importance for highly efficient protein adsorption and separation, yet still faces a huge challenge. Herein, a new kind of butane tetracarboxylic acid (BTCA) functionalized ethylene vinyl alcohol (EVAL) nanofibrous membranes (BTCA@EVAL NFM)-based chromatographic media is fabricated, for the first time, by combining blend electrospinning technique with in-situ modification technology. The resulting BTCA@EVAL NFM possesses an enhanced equilibrium protein adsorption capability (716 mg g−1), a high saturated dynamic protein binding capacity (490 mg g−1), and a distinctive selectivity towards positively charged proteins, which are attributed to the synergistic effects of the hydrophilic EVAL nanofibrous matrix and the plentiful carboxyl groups introduced by BTCA. Besides, benefiting from its stable physical and chemical structures, the membrane also presents excellent acid and alkaline resistance as well as good reusability. Significantly, the BTCA@EVAL NFM can directly extract lysozyme from egg white with a relatively large capture capability of 353 mg g−1, highlighting its superb potential practicability. We sincerely hope that this new design concept and the highly effective nanofiber-based chromatographic media can provide some guidance for the further development of bio-separation and purification.

Hydrophobic interaction chromatography (HIC) as an indispensable method for protein purification has attracted considerable attentions of researchers as well as biopharmaceutical industries. However, the low binding capacity and slow adsorption rate of the currently available HIC media lead to a little supply and high price of the highly purified proteins. Herein, nanofibrous membranes with hydrophobic binding sites were developed for HIC by directly coupling phenyl glycidyl ether on the hydrolyzed cellulose acetate nanofiber membrane (cellulose-phenyl NFM). Scanning electron microscope (SEM), water contact angle (WCA), Fourier transform infrared (FTIR), thermogravimetric analysis (TGA), Brunauer–Emmett–Teller (BET) surface area analysis and capillary flow porometer (CFP) were applied to evaluate the physically and chemically structural transformation. The obtained cellulose-phenyl NFMs showed a proper hydrophilcity (WCA = 37°), a relatively high BET surface area (3.6 times the surface area of commercial fibrous membranes), and tortuous-channel structure with through-hole size in the range of 0.25–1.2 μm, which led to a little non-specificity adsorption, high bovine serum albumin adsorption capacity of 118 mg g−1, fast adsorption process within 12 h, good long-term stability and reusability. Moreover, compared with traditional modification methods which always include activation and graft two steps, direct coupling method is more efficient for HIC media fabrication. Therefore, cellulose-phenyl NFMs with outstanding protein adsorption performance could be a kind of promising candidate for HIC.

The sulfur-coordinated organoiridium(III) complexes pbtIrSS and ppyIrSS, which contain C,N and S,S (dithione) chelating ligands, were found to inhibit breast cancer tumorigenesis and metastasis by targeting Wnt/β-catenin signaling for the first time. Treatment with pbtIrSS and ppyIrSS induces the degradation of LRP6, thereby decreasing the protein levels of DVL2, β-catenin and activated β-catenin, resulting in downregulation of Wnt target genes CD44 and survivin. Additionally, pbtIrSS and ppyIrSS can suppress cell migration and invasion of breast cancer cells. Furthermore, both complexes show the ability to inhibit sphere formation and mediate the stemness properties of breast cancer cells. Importantly, pbtIrSS exerts potent anti-tumor and anti-metastasis effects in mouse xenograft models through the blockage of Wnt/β-catenin signaling. Taken together, our results indicate that pbtIrSS has great potential to be developed as a breast cancer therapeutic agent with a novel mechanism.

Creating adsorptive materials for the fast, efficient, and high-throughput adsorption and purification of proteins is critical to meet the great demands for highly purified proteins, yet it has proven to be a highly challenging task. Here, we report that cross-linked and highly carboxylated poly(vinyl alcohol) (PVA) nanofibrous membranes were fabricated by a combination of electrospinning and the in situ graft polymerization of PVA and maleic anhydride (MAH) under mild conditions. Taking advantage of the large surface area available for protein binding, the highly tortuous porous structure, and the robust mechanical properties, the resultant PVA/MAH nanofibrous membranes exhibited a good integrated adsorption performance towards lysozyme, including a superior adsorption capacity of 177 mg g-1, fast adsorption equilibrium within 4 h, good selectivity, and good reversibility. Moreover, the saturation dynamic adsorption amount towards lysozyme reached 159 mg g-1 under 750 Pa driven solely by gravity, which conformed to the specified requirements for high adsorption capacity under relatively low pressure drops. Furthermore, the adsorption performance towards a protein mixture was analyzed by sodium dodecyl sulphate-polyacrylamide gel electrophoresis and the resultant PVA/MAH nanofibrous membranes retained excellent stability under depyrogenation conditions. The successful fabrication of such fascinating nanofibrous materials by using this simple and intriguing approach may provide new insights into the design and development of adsorptive materials for the purification of proteins with superior adsorption performance.

With the rapid urbanization and fast growth of transportation, noise pollution has become one of the most serious environmental problems in front of the people worldwide, it usually causes various disorders and greatly affects the work efficiency and living standards of human beings. Reducing noise by using sound absorption materials is an important approach to lessen the harm of noise pollution. As the most abundantly used materials, porous materials combine the properties of lightweight, wide absorption frequency range and highly sound absorption ability, and they hold great potential in the field of sound absorption. In this review, the recent progress in the design and fabrication of porous sound absorption materials is summarized and highlighted. This review covers the introduction of the sound absorption mechanism and evolution of prediction models for porous sound absorption materials, and the research and development of the design concepts and fabrication of sound absorption foams and fibrous sound absorption materials. The review concludes with some perspectives and outlook for the porous sound absorption materials.

Fabricating metal-chelating affinity materials with excellent protein adsorption and isolation properties is of great significance in numerous biotechnological fields, yet still face huge challenges. Herein, we successfully developed a new kind of nanofiber based metal-chelating affinity membranes by orderly modifying the electrospun regenerated cellulose nanofibrous membranes (RC NFMs) with intermediate bridging agent of cyanuric chloride (CC), chelating agent of iminodiacetic acid (IDA) and affinity ligand of Fe3+ ions. The resultant Fe3+ ions immobilized RC NFMs (RC-CC-IDA-Fe3+ NFMs) exhibited a high lysozyme adsorption amount of 365 mg g-1 within the fast equilibrium adsorption time of 4 hours. Meanwhile, the performance of RC-CC-IDA-Fe3+ NFMs could be facilely regulated by controlling the properties and concentration of protein solutions. Moreover, the obtained nanofibrous affinity materials also possessed superior cycle performance and excellent regeneration ability. Undoubtedly, the successful fabrication of efficient RC-CC-IDA-Fe3+ NFMs might provide a new kind of affinity adsorbent for protein purification and other potential biotechnological fields.

Highly effective separation and purification of protein are extremely important for the development of pharmaceutical industry, which still remains a great challenge. As a kind of frontier new material, electrospun nanofibers show vast perspectives in fabricating high performance protein adsorption materials attributing to their competitive structure advantages involving large specific surface area, high porosity, good pore channel connectivity, easy to surface functionalization, controllable single fiber and aggregate structure. Herein, we detailedly summarize the recent progress in the design concepts, synthesis and structure control of electrospun nanofibrous composite materials with different functional interfaces for the separation and purification of proteins. Furthermore, the current challenges and future development directions in this rapidly developed field are also highlighted in this review.

Construction ion-exchange membranes with superb biomolecules adsorption and purification performance plays a greatly important role in the fields of biotechnological and biopharmaceutical industry, yet still remains an extremely challenging. Herein, we in situ synthesized the cis-butenedioic anhydride grafted poly(vinyl alcohol) hydrogel nanofibrous membranes (CBA-g-PVA HNFM) by combining electrospinning technique with the grafting-copolymerization crosslinking. Taking advantages of the large specific surface area which could provide numerous sites available for functional groups and biomolecules binding, highly tortuous and interconnected porous channel for biomolecules transfer, and enhanced mechanical strength, the resultant CBA-g-PVA HNFM exhibited relatively high binding amount of 170 mg g−1, rapid equilibrium time of 8 h towards the biomolecule template of lysozyme, and the performance could be tailored by regulating the buffer properties and protein concentrations. Additionally, the resultant functional HNFM also possessed superior acid resistance property, excellent reversibility and regeneration performance. More importantly, the obtained CBA-g-PVA HNFM could directly extract lysozyme from fresh chicken eggs with capacity of 125 mg g−1, exhibiting excellent practical application properties. The fabrication of proposed CBA-g-PVA HNFM offers a feasible alternative for construction of ion-exchange chromatograph column for bio-separation and purification engineering.

Developing nanofibrous aerogels with high porosity, robust underwater mechanical strength, and rich adsorption ligands, has been considered as one of the most promising strategies for preparing the next generation of high-efficiency and high-throughput chromatographic media; yet great challenges still remain. Herein, a novel type of highly phosphorylated nanofibrous aerogels (PNFAs) is fabricated, for the first time, by combining electrospinning, cryogenic induced phase separation regulation, and in situ phosphorylation modification. The PNFAs exhibit outstanding underwater superelasticity and excellent compression fatigue resistance (∼0% plastic deformation after 1000 compression cycles), as well as favorable shape-memory property. Besides, the PNFAs also can be bent and compressed even in the ultracold liquid nitrogen without obvious plastic deformation, further highlighting their robust structural stability. Benefiting from the superelastic, interconnected, and highly phosphorylated 3D nanofibrous frameworks, the PNFAs possess a superb protein adsorption capability of 3.3 × 103 mg g-1 and a large liquid flux of 1.5 × 104 L m-2 h-1, which are superior to the commercial and previously reported fiber-based chromatographic media. Moreover, the PNFAs also exhibit superior performance stability, easy assembly, and outstanding applicability, highlighting their potential actual application. The successful preparation of such fascinating PNFAs may not only provide a new option for the current protein adsorption and purification engineering, but also could open up some new perspectives for further design and development of next-generation nanofibrous aerogel-based chromatographic media for various bioseparation applications.

Effective separation of oils from water surfaces and oil/water mixtures is crucial for oil purification and oily wastewater treatment, and remains a worldwide challenge. Electrospun nanofibers are considered a promising candidate for oil–water separation, with competitive advantages including a large specific surface area, high porosity, good pore channel connectivity, facile tunability of surface wettability, and easily scalable fabrication from various materials. According to the properties of oil/water mixtures (immiscible or emulsified), various electrospun nanofibrous materials in the form of bulky mats, membranes, and aerogels have been developed; meanwhile, special surface wettability including superhydrophobic–superoleophilic, superhydrophilic–superoleophobic, and stimulus–responsive switchable wettability have been designed for oil–water separation in different conditions. This chapter reviews recent achievements in the development of electrospun nanofibrous materials for oil–water separation applications. The current challenges and promising prospects in this rapidly developing field are also considered.

// Qiuxia Fu 1, * , Shaoduo Yan 1, * , Licui Wang 1 , Xiangguo Duan 2 , Lei Wang 1 , Yue Wang 1 , Tao Wu 3 , Xiaohui Wang 1 , Jie An 1 , Yulong Zhang 1 , Qianqian Zhou 1 , Linsheng Zhan 1 1 Beijing Institute of Transfusion Medicine, Beijing Key Laboratory of Blood Safety and Supply Technologies, Beijing, China 2 Surgical Laboratory of General Hospital, Ningxia Medical University, Yinchuan, China 3 Blood Transfusion Department, General Hospital of Beijing Military Area Command of PLA, Beijing, China * These authors have contributed equally to this work Correspondence to: Linsheng Zhan, email: lszhan91@yahoo.com Keywords: HCV, codon-optimized φC31 integrase, NK cells, liver injury Received: December 13, 2015 Accepted: July 18, 2016 Published: August 04, 2016 ABSTRACT The role of hepatic NK cells in the pathogenesis of HCV-associated hepatic failure is incompletely understood. In this study, we investigated the effect of HCV on ConA-induced immunological hepatic injury and the influence of HCV on hepatic NK cell activation in the liver after ConA administration. An immunocompetent HCV mouse model that encodes the entire viral polyprotein in a liver-specific manner based on hydrodynamic injection and φC31o integrase was used to study the role of hepatic NK cells. Interestingly, the frequency of hepatic NK cells was reduced in HCV mice, whereas the levels of other intrahepatic lymphocytes remained unaltered. Next, we investigated whether the reduction in NK cells within HCV mouse livers might elicit an effect on immune-mediated liver injury. HCV mice were subjected to acute liver injury models upon ConA administration. We observed that HCV mice developed more severe ConA-induced immune-mediated hepatitis, which was dependent on the accumulated intrahepatic NK cells. Our results indicated that after the administration of ConA, NK cells not only mediated liver injury through the production of immunoregulatory cytokines (IFN-γ, TNF-α and perforin) with direct antiviral activity, but they also killed target cells directly through the TRAIL/DR5 and NKG2D/NKG2D ligand signaling pathway in HCV mice. Our findings suggest a critical role for NK cells in oversensitive liver injury during chronic HCV infection.

With the objective of investigating the acute activation of inflammatory cascades upon exposure to gold nanoparticles (GNPs) as well as detailing the mechanisms, a reporter mouse model that allows for non-invasive and longitudinal imaging of hepatic acute-phase serum amyloid A (SAA) activation is constructed. The model is able to visualize SAA activation at the transcriptional stage, with higher sensitivity than serum protein detection by ELISA. GNPs of various sizes (10-80 nm) and geometries are assessed using the reporter mice with results demonstrating that 50 nm nanospheres (GNS50) possess the highest capacity to induce hepatic SAA activation. Detailed analysis uncovers that resident macrophages in the liver are the main origins of these cytokines and that the exposure to GNS50 significantly induces the M1 macrophage phenotype. Moreover, those M1-polarized macrophages, together with the subsequently secreted pro-inflammatory cytokines, exert effects on hepatocytes and then initiate SAA transcription through the NF-κB signal pathway. The results detail the sequential reactions to GNPs among macrophages, inflammatory mediators, and SAA-synthesizing hepatocytes, which shed light on the acute effects of GNPs on the body. In addition, the established in situ and highly sensitive SAA detection system is expected to have vast applications in evaluating NP-induced acute inflammatory reactions.

Fabricating protein adsorbents with high adsorption capacity and appreciable throughput is extremely important and highly desired for the separation and purification of protein products in the biomedical and pharmaceutical industries, yet still remains a great challenge. Herein, we demonstrate the synthesis of a novel protein adsorbent by in situ functionalizing eletrospun ethylene-vinyl alcohol (EVOH) nanofibrous membranes (NFM) with critic acid (CCA). Taking advantage of the merits of large specific surface area, highly tortuous open-porous structure, abundant active carboxyl groups introduced by CCA, superior chemical stability, and robust mechanical strength, the obtained CCA-grafted EVOH NFM (EVOH-CCA NFM) present an excellent integrated protein (take lysozyme as the model protein) adsorption performance with a high capacity of 284 mg g(-1), short equilibrium time of 6 h, ease of elution, and good reusability. Meanwhile, the adsorption performance of EVOH-CCA NFM can be optimized by regulating buffer pH, ionic strength, and initial concentration of protein solutions. More importantly, a dynamic binding efficiency of 250 mg g(-1) can be achieved driven solely by the gravity of protein solution, which matches well with the demands of the high yield and energy conservation in the actual protein purification process. Furthermore, the resultant EVOH-CCA NFM also possess unique selectivity for positively charged proteins which was confirmed by the method of sodium dodecyl sulfate polyacrylamide gel electrophoresis. Significantly, the successful synthesis of such intriguing and economic EVOH-CCA NFM may provide a promising candidate for the next generation of protein adsorbents for rapid, massive, and cost-effective separation and purification of proteins.

Daylight-driven antimicrobial nanomaterials serve as a novel barrier for transmission of emerging infectious diseases., Emerging infectious diseases (EIDs) are a significant burden on global economies and public health. Most present personal protective equipment used to prevent EID transmission and infections is typically devoid of antimicrobial activity. We report on green bioprotective nanofibrous membranes (RNMs) with rechargeable antibacterial and antiviral activities that can effectively produce biocidal reactive oxygen species (ROS) solely driven by the daylight. The premise of the design is that the photoactive RNMs can store the biocidal activity under light irradiation and readily release ROS under dim light or dark conditions, making the biocidal function “always online.” The resulting RNMs exhibit integrated properties of fast ROS production, ease of activity storing, long-term durability, robust breathability, interception of fine particles (>99%), and high bactericidal (>99.9999%) and virucidal (>99.999%) efficacy, which enabled to serve as a scalable biocidal layer for protective equipment by providing contact killing against pathogens either in aerosol or in liquid forms. The successful synthesis of these fascinating materials may provide new insights into the development of protection materials in a sustainable, self-recharging, and structurally adaptive form.

Construction of adsorptive materials for simple, efficient, and high-throughput adsorption of proteins is critical to meet the great demands of highly purified proteins in biotechnological and biopharmaceutical industry; however, it has proven extremely challenging. Here, we report a cost-effective strategy to create carbonyl groups surface-functionalized nanofibrous membranes under mild conditions for positively charged protein adsorption. Our approach allows maleic anhydride to in situ graft on cellulose nanofibrous membranes (CMA) to construct adsorptive membranes with large surface area and tortuous porous structure. Thereby, the resultant CMA membranes exhibited high adsorption capacity of 160 mg g(-1), fast equilibrium within 12 h, and good reversibility to lysozyme. Moreover, the dynamic adsorption was performed under low pressure-drops (750 Pa), with a relatively high saturation adsorption amount of 118 mg g(-1), which matched well with the requirements for proteins purification. Considering the excellent adsorption performance of the as-prepared adsorptive membranes, this simple and intriguing approach may pave a way for the design and development of robust and cost-effective adsorption membranes to meet the great demands for fast and efficient adsorption of positively charged proteins.

Many applications proposed for functional nanofibers require their assembly into a monolithic cellular structure. The ability to maintain structural integrity upon large deformation is essential to ensure a macroscopic cellular material that functions reliably. However, it remains a great challenge to achieve high elasticity in three-dimensional (3D) nanofibrous networks. Here, we report a strategy to create fibrous, isotropically bonded elastic reconstructed (FIBER) aerogels with a hierarchical cellular structure and superelasticity by combining electrospun nanofibers and the freeze-shaping technique. Our approach allows the intrinsically lamellar deposited electrospun nanofibers to assemble into elastic bulk aerogels with tunable porous structure and wettability on a large scale. The resulting FIBER aerogels exhibit the integrated properties of ultralow density (30 mg cm(-3)), rapid recovery from 80% compression strain, superhydrophobic-superoleophilic wettability, and high pore tortuosity. More interestingly, the FIBER aerogels can effectively separate surfactant-stabilized water-in-oil emulsions, solely using gravity, with high flux (maximum of 8140 ± 220 L m(-2) h(-1)) and high separation efficiency, which match well with the requirements for treating the real emulsions. The synthesis of FIBER aerogels also provides a versatile platform for exploring the applications of nanofibers in a self-supporting, structurally adaptive, and 3D macroscopic form.

Apoptosis is an essential process for the maintenance of liver physiology. The ability to noninvasively image apoptosis in livers would provide unique insights into its role in liver disease processes. In the present work, we established a stable mouse model by hydrodynamics methods to study the activity of caspase-3 and evaluate the effect of the apoptosis inhibitors in mouse livers under true physiological conditions by bioluminescence imaging. The reporter plasmid attB-ANLuc(DEVD)BCLuc that contains fragment of attB and ANLuc(DEVD)BCLuc was codelivered with the mouse-codon optimized φC31 (φC31o) integrase plasmids specifically to mouse liver by hydrodynamic injection procedure. Then, φC31o integrase mediated intramolecular recombination between wild-type attB and attP site in mice, and thus the reporter expression cassette attB-ANLuc(DEVD)BCLuc was integrated permanently into mouse liver chromosome. We used these mice to characterize in vivo activation of caspase-3 upon treatment with LPS/D-GalN. Our data show that liver apoptosis could be reflected by the activity of luciferase. The shRNA targeting caspase-3 protein or apoptosis inhibitors could effectively downregulate luciferase activity in vivo. Also, this model could be used to measure caspase-3 activation during inflammatory and infectious events in vivo as verified by infected with MHV-3. This model could be used for screening anti-apoptosis compounds target mouse livers.

The lack of efficient antigen and activator delivery systems, as well as the restricted migration of dendritic cells (DCs) to secondary lymph organs, dramatically limits DC-based adoptive immunotherapy. We selected two spherical gold nanoparticle (AuNP)-based vehicles of optimal size for activator and antigen delivery. Their combination (termed the NanoAu-Cocktail) was associated with the dual targeting of CpG oligonucleotides (CpG-ODNs) and an OVA peptide (OVAp) to DC subcellular compartments, inducing enhanced antigen cross-presentation, upregulated expression of costimulatory molecules and elevated secretion of T helper1 cytokines. We demonstrated that the intravenously transfused NanoAu-Cocktail pulsed DCs showed dramatically improved in vivo homing ability to lymphoid tissues and were settled in T cell area. Especially, by tissue-distribution analysis, we found that more than 60% of lymphoid tissues-homing DCs accumulated in liver-draining lymph nodes (LLNs). The improved homing ability of NanoAu-Cocktail pulsed DCs was associated with the high expression of chemokine receptor 7 (CCR7) and rearrangement of the cytoskeletons. In addition, by antigen-specific tetramers detection, NanoAu-Cocktail pulsed DCs were proved able to elicit strong antigen-specific CD8+ T cell responses, which provided enhanced protection from viral invasions. This study highlights the importance of codelivering antigen/adjuvant using different sized gold nanoparticles to improve DC homing and therapy.

In the current work, a one-step, washing-free, homogeneous nanosensor assay has been constructed to sensitively detect hepatitis B surface antigen (HBsAg) based on the light scattering property of gold nanoparticles (GNPs) through a sandwich model. The two nanoprobes in this study were designed by conjugating monoclonal and polyclonal hepatitis B surface antibody (HBsAb) onto the GNPs of different diameters. First, the detection behavior of the combinations of different sizes of GNPs was evaluated and the optimized combination was determined. In analyzing HBsAg in Tris-HCl buffer, such bioassay composed of GNPs of approximately 50 and 100 nm has a limit of detection (LOD) as high as 0.005 IU/ml and a dose-dependent response ranging from 0.005 to 1 IU/ml, which indicates its good diagnostic capability and provides a useful means to analyze protein biomarkers with low virus loads. Observation with transmission electron microscopy (TEM) provides direct evidence that the increase of hydrodynamic diameters resulted from the aggregation induced by immunological reactions. The bioassay also exhibits satisfactory specificity in analyzing HBsAg in serum media. Therefore, with its simple preparation, easy readout, and good stability, this bioassay has the potential to be developed into an automated and widely used biosensor assay.

Hydrodynamic-based gene delivery has emerged as an efficient and simple method for the intracellular transfection of naked plasmid DNA (pDNA) in vivo. In this system, a hydrodynamic injection via the tail vein is the most effective non-viral method of liver-targeted gene delivery. However, this injection is often technically challenging when used in animals whose tail veins are difficult to visualize or too small to operate on. To overcome this limitation, an alternative in vivo gene delivery method, the rapid injection of large volume of pDNA solution through retro-orbital sinus, was established. Using this technique, we successfully delivered pDNA to the tissue of adult mice, neonatal mice and tree shrews. The efficient expression of exogenous genes was specifically detected in the liver of test animals treated with this gene delivery method. This study demonstrates for the first time that the hydrodynamic gene delivery via the retro-orbital sinus can not only reach the same transgene efficiency as a tradition hydrodynamic-based intravascular injection but also be used in animals that are difficult to inject via the tail vein. This method could open up new areas in gene function studies and gene therapy disease treatment.

A novel gold nanorods (GNRs) biosensor based on the localized surface plasmon resonance (LSPR) behavior was designed to detect the hepatitis B surface antigen (HBsAg) which indicates active viral replication of hepatitis B virus (HBV). The surface of GNRs was modified with monoclonal hepatitis B surface antibody (HBsAb) through physical adsorption. Raman spectrum, dynamic light scattering (DLS) and zeta potential measurement were conducted to access the nature of the GNRs after antibody modification. The binding of analyte to the molecular probe was monitored by the longitudinal wavelength shift of LSPR peak in the UV-Vis extinction spectrum resulting from the changes of local refractive index induced by the immunological reaction. The biosensor could be utilized in quantitative analysis in Tris buffers, which has dose-dependence response ranging from 0.01 IU/mL to 1 IU/mL. Further, the biosensor was suited for qualitative analysis of HBsAg in the actual media of blood serum and plasma. The ease of operation, high sensitivity, and its generality offer specific advantages over other GNRs-based immunoassay methods.

Nanofiber-based hydrogels (NFHGs) prepared by the combination of traditional hydrogels and novel nanofibers have demonstrated great potential in various application fields, owing to their integrated advantages of superhydrophilicity, high water-holding capacity, good biocompatibility, enhanced mechanical strength, and excellent structural tenability. In this review, a comprehensive overview of the structure design and synthetic strategy of NFHGs derived from electrospinning technique, weaving, freeze-drying, 3D printing, and molecular self-assembling method is provided. The widely researched multifunctional applications, primarily involving tissue engineering, drug delivery, sensing, intelligent actuator, and oil/water separation are also presented. Furthermore, some unsolved scientific issues and possible directions for future development of this field are also intensively discussed.

To improve the efficiency of expression of reporter transgenes delivered by hydrodynamic injection, we generated expression cassettes carrying different liver-specific regulatory elements using the firefly luciferase gene as a reporter. From our studies the human alpha-antitrypsin promoter together with the apolipoprotein E/C-I and albumin enhancer was the combination of choice for prolonged transgene expression, but reporter gene expression in vivo lasted for no more than 7 weeks. Subsequently, phage phiC31 integrase was introduced as a potential tool to improve further the efficiency of expression of the delivered transgene. Long-term transgene expression in vivo was achieved by specific integration of the target gene into mouse livers. This study demonstrates the use of a combination of phage phiC31 integrase and liver-specific regulatory elements for generation of transgenic mice.

Background The lack of a robust small animal model for hepatitis C virus (HCV) has hindered the development of novel drugs, including internal ribosome entry site (IRES) inhibitors. Phage φC31 integrase has emerged as a potent tool for achieving long-term gene expression in vivo. This study utilized φC31 integrase to develop a stable, reproducible and easily accessible HCV IRES mouse model. Methods φC31 integrase plasmid and the reporter vector, HCV-IRES–luciferase expression cassette (containing an attB site), was codelivered to murine livers using high pressure tail vein injection. HCV IRES-dependent translation refected by luciferase expression was accurately monitored in vivo by bioluminescence imaging. Genomic integration of the transgene was confirmed by partial hepatectomy and nested PCR. An HCV IRES-targeted short hairpin RNA (shRNA) expression plasmid, sh184, was hydrodynamically transfected into mouse liver to study its inhibition efficacy in vivo. Results φC31 integrase mediated intramolecular recombination between wild-type attB and attP sites in mice. The expression of luciferase was stable after 30 days post-transfection and remained so for 300 days only in the livers of mice that were coinjected with the integrase-encoding plasmid. Luciferase levels reduced dramatically after hydrodynamic transfection of sh184. Conclusions These results indicate that this mouse model provides a powerful tool for accurate and long-term evaluation of potential anti-IRES compounds in vivo.

Novel flexible, thermally stable and hierarchical porous silica nanofibrous membranes with superhydrophilicity and underwater superoleophobicity were prepared by a facile in situ synthesis method, which can effectively separate oil-in-water microemulsions solely driven by gravity, with an extremely high flux of 2237 L m(-2) h(-1).

The nuclear factor-κB (NF-κB) signaling pathway plays a critical role in a multitude of cellular processes. Activation of the NF-κB transcription factor family is essential for the initiation of inflammation, immunity, cell proliferation and apoptosis through a list of responsive genes. In hepatic tissue, activation of the NF-κB pathway has been implicated in a number of pathological conditions. Here we described a mouse model for noninvasive quantification of NF-κB activation in the hepatic tissues. Mice were subjected to hydrodynamic delivery with a mixture of pattB-NF-κB-Fluc reporter and φC31o integrase vector. Hepatic expression of φC31o integrase mediated chromosomal integration of the pattB-NF-κB-Fluc reporter, resulting in stable luciferase expression at 300 days post transfection. We applied noninvasive imaging and were able to detect NF-κB activation under acute liver injury and hepatitis conditions. During hepatectomy-induced liver regeneration, NF-κB activation was detected locally in the tissues at the surgery site. Treatment with Sorafenib suppressed NF-κB activation, accompanied with perturbation of liver regeneration. In conclusion, we established a method for stable transfection of the hepatic tissues and applied the transfected mice to longitudinal monitoring of NF-κB activity under pathological conditions. Further exploration of this methodology for establishment of other disease models and for evaluation of novel pharmaceuticals is likely to be fruitful.

Hepatitis B virus(HBV) infection remains a global problem, despite the effectiveness of the Hepatitis B vaccine in preventing infection. The resolution of Hepatitis B virus infection has been believed to be attributable to virus-specific immunity. In vivo direct evaluation of anti-HBV immunity in the liver is currently not possible. We have developed a new assay system that detects HBV clearance in the liver after the hydrodynamic transfer of a reporter gene and over-length, linear HBV DNA into hepatocytes, followed by bioluminescence imaging of the reporter gene (Fluc). We employed bioluminescence detection of luciferase expression in HBV-infected hepatocytes to measure the Hepatitis B core antigen (HBcAg)-specific immune responses directed against these infected hepatocytes. Only HBcAg-immunized, but not mock-treated, animals decreased the amounts of luciferase expression, HBsAg and viral DNA from the liver at day 28 after hydrodynamic infection with over-length HBV DNA, indicating that control of luciferase expression correlates with viral clearance from infected hepatocytes.

It remains challenging to detect unknown pathogenic bacteria in diagnostic, clinical and environmental fields. This work describes the approach to the development of a sensitive, broad-range genosensing assay targeting the conserved 16S rDNA region existing in most bacteria, by monitoring the aggregation level of gold nanorods (GNRs)-based nanoprobes through their localized surface plasmon resonance (LSPR) property. In the quantitative detection of artificial sequence, the limit of detection (LOD) of such a bioassay is demonstrated to reach the 5 pM level. This pair of universal GNRs-based nanoprobes can further identify at least 6 species of bacteria that were most prevalent in platelet concentrates (PCs) and have no cross-reaction with other pathogens. Moreover, it also exhibits higher sensitivity than other broad-range methods in analysing Serratia marcescens-spiked PCs. Therefore, the presented strategy not only provides a novel and effective DNA analysis method to detect multiple bacterial contaminations in PCs, but also opens up possibilities for its future use of detecting unknown bacteria in other systems, such as food and water, even at ultralow levels.

In the present study, we aimed to develop a nucleic acid lateral-flow method for the rapid and sensitive detection of multiple bacteria that contaminate platelet concentrations (PCs). Polymerase chain reaction (PCR) amplicons were produced by a set of board-range primers that recognize the conserved region of bacteria 16S rDNA, followed by hybridization with both an FITC (fluorescein isothiocyanate)-labelled probe and biotin-labelled probe, and then a nucleic acid lateral-flow dipstick (LFD) assay. The LFD accurately identified 7 species of bacteria, but had no cross-reactivity with human genomic DNA. The limit of detection (LOD) of the LFD assay was as low as 10(1) copies/µL of 16S rDNA for plasmid. In the case of spiked PCs without enrichment, the detection limit of LFD for K. pneumonia was 5 CFU/mL, 6.5 × 10(4) CFU/mL for the S. epidermidis and 35 CFU/mL for P. aeruginosa.

Background/Aims Interferon beta (IFN-β) is the priming cytokine in the interferons (IFNs) response that plays essential roles in innate immune system. Only very few studies on IFN activation in animals have been reported before, therefore, we embarked to develop a novel method to dynamically examine IFN-β activation in mouse liver by noninvasive molecular imaging. Methods Interferon beta promoter-directed firefly luciferase gene was integrated into chromosomes of hepatocytes by hydrodynamic injection. Mouse hepatitis virus type 3 (MHV-3) and polyinosinic-polycytidylic acid [poly(I:C)] were used to stimulate the activation of IFN-β. Luciferase activity was used as an indicator of the IFN-β promoter activity in vitro and in vivo. The expression level of IFN-β in the sera and firefly luciferase in the liver was assessed by ELISA and bioluminescence imaging respectively. Western blot was used for detecting proteins expression. Results A rapid and elevated luciferase expression in the mouse liver induced by poly (I:C) and MHV-3 was detected by bioluminescence imaging. The detectable level of IFN-β in the sera was not induced by MHV-3. Moreover, IFN-β activation was significantly inhibited by the hepatitis C virus (HCV) NS3/4A protease in mouse liver. These results were consistent with IFN-β production in the sera. Therefore, a novel visual method to monitor IFN-β promoter activity was established in the current study. Conclusion This novel sensitive method can be used for not only assessing IFN-β activation or inhibition in the liver under different conditions, but also screening drug candidates of stimulating or inhibiting of IFN-β production.

The identification and quantitative analysis of protein-protein interactions are essential to the functional characterization of proteins in the post-proteomics era. The methods currently available are generally time-consuming, technically complicated, insensitive and/or semi-quantitative. The lack of simple, sensitive approaches to precisely quantify protein-protein interactions still prevents our understanding of the functions of many proteins. Here, we develop a novel dual luciferase reporter pull-down assay by combining a biotinylated Firefly luciferase pull-down assay with a dual luciferase reporter assay. The biotinylated Firefly luciferase-tagged protein enables rapid and efficient isolation of a putative Renilla luciferase-tagged binding protein from a relatively small amount of sample. Both of these proteins can be quantitatively detected using the dual luciferase reporter assay system. Protein-protein interactions, including Fos-Jun located in the nucleus; MAVS-TRAF3 in cytoplasm; inducible IRF3 dimerization; viral protein-regulated interactions, such as MAVS-MAVS and MAVS-TRAF3; IRF3 dimerization; and protein interaction domain mapping, are studied using this novel assay system. Herein, we demonstrate that this dual luciferase reporter pull-down assay enables the quantification of the relative amounts of interacting proteins that bind to streptavidin-coupled beads for protein purification. This study provides a simple, rapid, sensitive, and efficient approach to identify and quantify relative protein-protein interactions. Importantly, the dual luciferase reporter pull-down method will facilitate the functional determination of proteins.

AIM: To develop a sensitive assay for screening compounds against hepatitis C virus (HCV). METHODS: The proteolytic cleavage of NS3/4A on enhanced yellow fluorescent protein (eYFP)-mitochondrial antiviral signaling protein (MAVS) was examined by reporter enzyme secreted placental alkaline phosphatase (SEAP), which enabled us to perform ongoing monitoring of anti-HCV drugs through repeated chemiluminescence. Subcellular localization of eYFP-MAVS was assessed by fluorescence microscopy. Cellular localization and protein levels were examined by Western blotting. RESULTS: HCV NS3/4A protease cleaved eYFP-MAVS from mitochondria to block the activation of interferon (IFN)-β promoter, thus resulting in downregulation of SEAP activity. The decrease in SEAP activity was proportional to the dose of active NS3/4A protease. Also this reporter assay was used to detect anti-HCV activity of IFN-α and cyclosporine A. CONCLUSION: Our data show that this reporter system is a sensitive and quantitative reporter of anti-HCV inhibitors. This system will constitute a new tool to allow the efficient screening of HCV inhibitors.