ROS-responsive nano-medicine for navigating autophagy to enhance targeted therapy of inflammatory bowel disease.

Inflammatory bowel disease (IBD) is a chronic gastrointestinal disorder characterized by immune dysregulation and intestinal inflammation. Rapamycin (Ra), an mTORC1 pathway inhibitor, has shown promise for autophagy induction in IBD therapy but is associated with off-target effects and toxicity. To address these issues, we developed an oral liposome responsive to reactive oxygen species (ROS) using lipids and amphiphilic materials. We combined ketone thiol (TK) for ROS responsive and hyaluronic acid (HA) with high affinity for CD44 receptors to prepare rapamycin-loaded nanoparticle (Ra@TH). Owing to its ROS responsive characteristic, Ra@TH can achieve inflammatory colonic targeting. Additionally, Ra@TH can induce autophagy by inhibiting the mTORC1 pathway, leading to the clearance of damaged organelles, pathogenic microorganisms and oxidative stress products. Simultaneously, it also collaboratively inhibits the NF-κB pathway suppressed by the removal of ROS resulting from TK cleavage, thereby mediating the expression of inflammatory factors. Furthermore, Ra@TH enhances the expression of typical tight junction proteins, synergistically restoring intestinal barrier function. Our research not only expands the understanding of autophagy in IBD treatment but also introduces a promising therapeutic approach for IBD patients.

Sandwich-type electrochemical aptasensor for sensitive detection of myoglobin based on Pt@CuCo-oxide nanoparticles as a signal marker.

When an acute myocardial infarction (AMI) occurs, myoglobin (Mb) is the biomarker whose concentration firstly increases, and the high sensitive detection of Mb is critical for early diagnosis of AMI. Herein, a sandwich-type electrochemical aptasensor for the sensitive detection of Mb was constructed by using Pt@Cu1.33OCo0.83O as the signal marker. On one hand, nano-flower-like Cu1.33OCo0.83O was synthesized by hydrothermal method and Pt nanoparticles (Pt NPs) were loaded on its surface. Pt@Cu1.33OCo0.83O could immobilize aptamer 2 (Apt2) successfully by the Pt-S bond. And because of the synergistic effect between Pt and bimetallic oxide, Pt@Cu1.33OCo0.83O had an excellent catalytic effect on the signal source of hydrogen peroxide (H2O2) to amplify the current signal, which enhance the sensitivity of the aptasensor. On the other hand, the screen-printed gold electrode (SPGE) was used as the sensing base, which had good conductivity and ensured the immobilization of aptamer 1 (Apt1). The quantitative detection of Mb was achieved by specific recognition between Mb and Apt1, Apt2. As a result, the constructed electrochemical aptasensor had a good linear range (1-1500 ng/mL) with a low detection limit (LOD) of 0.128 ng/mL (S/N = 3), and a high sensitivity of 29.47 µA dec-1. The aptasensor also realized the detection of Mb in human serum samples with good accuracy, and the results were consistent with the hospital's biochemical indicators, which demonstrated the potential application of the prepared sensor in the clinical detection of Mb.

Multiply-mesoporous hydrophilic titanium dioxide nanohybrid for the highly-performed enrichment of N-glycopeptides from human serum.

N-glycopeptide is considered as one of significant biomarkers which provide guidance for the diagnosis and drug design of diseases. However, the direct analysis of N-glycopeptides is nearly impracticable mainly owing to their extremely low abundance and grave signal suppression from other interfering substances in the bio-samples. In this research, a multiply-mesoporous hydrophilic TiO2 nanohybrid (mM-TiO2@Cys) was synthesized by immobilizing Cys on a TiO2 substrate with hierarchical mesopores to achieve the highly-performed enrichment of N-glycopeptides. With the advantages of superior hydrophilicity and multiply-mesoporous structure, the obtained material exhibited an excellent selectivity (IgG digests and BSA digests at the molar ratio of 1/500), a high sensitivity (1 fmol µL-1 for IgG digests) and a good size-exclusion ability (IgG digests, IgG and BSA at the molar ratio of 1/500/500) in the enrichment of N-glycopeptides from IgG digests. As a result, 281 N-glycopeptides corresponded with 109 glycoproteins were identified from 2 µL serum digests of the patients with nasopharyngeal carcinoma, and 181 N-glycopeptides corresponded with 78 glycoproteins were identified from 2 µL serum digests of the healthy volunteers, revealing the potential application value of mM-TiO2@Cys in glycoproteomics.

Spatiotemporally controlled Pseudomonas exotoxin transgene system combined with multifunctional nanoparticles for breast cancer antimetastatic therapy.

The tumor microenvironment is a barrier to breast cancer therapy. Cancer-associated fibroblast cells (CAFs) can support tumor proliferation, metastasis, and drug resistance by secreting various cytokines and growth factors. Abnormal angiogenesis provides sufficient nutrients for tumor proliferation. Considering that CAFs express the sigma receptor (which recognizes anisamide, AA), we developed a CAFs and breast cancer cells dual-targeting nano drug delivery system to transport the LightOn gene express system, a spatiotemporal controlled gene expression consisting of a light-sensitive transcription factor and a specific minimal promoter. We adopted RGD (Arg-Gly-Asp) to selectively bind to the αvß3 integrin on activated vascular endothelial cells and tumor cells. After the LightOn system has reached the tumor site, LightOn gene express system can spatiotemporal controllably express toxic Pseudomonas exotoxin An under blue light irradiation. The LightOn gene express system, combined with multifunctional nanoparticles, achieved high targeting delivery efficiency both in vitro and in vivo. It also displayed strong tumor and CAFs inhibition, anti-angiogenesis ability and anti-metastasis ability, with good safety. Moreover, it improved survival rate, survival time, and lung metastasis rate in a mouse breast cancer model. This study proves the efficacy of combining the LightOn system with targeted multifunctional nanoparticles in tumor and anti-metastatic therapy and provides new insights into tumor microenvironment regulation.

Sequential Rocket-Mode Bioactivating Ticagrelor Prodrug Nanoplatform Combining Light-Switchable Diphtherin Transgene System for Breast Cancer Metastasis Inhibition.

The increased risk of breast cancer metastasis is closely linked to the effects of platelets. Our previously light-switchable diphtheria toxin A fragment (DTA) gene system, known as the LightOn system, has demonstrated significant therapeutic potential; it lacks antimetastatic capabilities. In this study, we devised an innovative system by combining cell membrane fusion liposomes (CML) loaded with the light-switchable transgene DTA (pDTA) and a ticagrelor (Tig) prodrug. This innovative system, named the sequential rocket-mode bioactivating drug delivery system (pDTA-Tig@CML), aims to achieve targeted pDTA delivery while concurrently inhibiting platelet activity through the sequential release of Tig triggered by reactive oxygen species with the tumor microenvironment. In vitro investigations have indicated that pDTA-Tig@CML, with its ability to sequentially release Tig and pDTA, effectively suppresses platelet activity, resulting in improved therapeutic outcomes and the mitigation of platelet driven metastasis in breast cancer. Furthermore, pDTA-Tig@CML exhibits enhanced tumor aggregation and successfully restrains tumor growth and metastasis. It also reduces the levels of ADP, ATP, TGF-ß, and P-selectin both in vitro and in vivo, underscoring the advantages of combining the bioactivating Tig prodrug nanoplatform with the LightOn system. Consequently, pDTA-Tig@CML emerges as a promising light-switchable DTA transgene system, offering a novel bioactivating prodrug platform for breast cancer treatment.

Polydopamine-coated ferric oxide nanoparticles for R848 delivery for photothermal immunotherapy in breast cancer.

Breast cancer, which requires comprehensive multifunctional treatment strategies, is a major threat to the health of women. To develop multifunctional treatment strategies, we combined photothermal therapy (PTT) with immunotherapy in multifunctional nanoparticles for enhancing the anti-tumor efficacy. Fe3O4 nanoparticles coated with the polydopamine shell modified with polyethylene glycol and cyclic arginine-glycyl-aspartic peptide/anisamide (tNP) for loading the immune adjuvant resiquimod (R848) (R848@tNP) were developed in this research. R848@tNP had a round-like morphology with a mean diameter of 174.7 ± 3.8 nm, the zeta potential of -20.9 ± 0.9 mV, the drug loading rate of 9.2 ± 1.1 %, the encapsulation efficiency of 81.7 ± 3.2 %, high photothermal conversion efficiency and excellent magnetic properties in vitro. Furthermore, this research also explored the anticancer efficacy of nanoparticles against the breast cancer under the near-infrared (NIR) light (808 nm) in vitro and in vivo. R848@tNP-based NIR therapy effectively inhibited the proliferation of breast cancer cells. Moreover, R848@tNP mediated PTT significantly enhanced the maturation of dendritic cells in vitro. Additionally, R848@tNP enhances the anti-tumor effect and evoked an immune response under NIR in vivo. Furthermore, the biosafety of R848@tNP was fully investigated in this study. Collectively, these results clearly demonstrate that R848@tNP, with magnetic resonance imaging characteristics, is a potential therapeutic for breast cancer that combines PTT with the immunotherapy.

Multiplexed detection of biomarkers using a microfluidic chip integrated with mass-producible micropillar array electrodes.

Quantifying multiple biomarkers with high sensitivity in tiny biological samples is essential to meet the growing demand for point-of-care testing. This paper reports the development of a novel microfluidic device integrated with mass-producible micropillar array electrodes (µAEs) for multiple biomarker detections. The µAE are mass-fabricated by soft lithography and hot embossing technique. Pt-Pd bimetallic nanoclusters (BNC) are modified on the surface of µAEs by constant potential (CP)/multi-potential step (MPS) electrodeposition strategies to improve the electroanalytical performance. The experimental result displays that Pt-Pd BNC/µAEs have good sensitivity enhancement compared with bare planar electrodes and bare µAEs, the enhancement being 56.5 and 9.5 times respectively, from the results of the H2O2 detection. Furthermore, glucose, uric acid and sarcosine were used as model biomarkers to show the biosensing capability with high sensitivity. The linear range and LOD of the glucose, uric acid and sarcosine detection are 0.1 mM-12 mM, 10 µM-800 µM and 2.5 µM-100 µM, 58.5, 3.4 and 0.4 µM, respectively. In particular, biosensing chips show wide linear ranges covering required detection ranges of glucose, uric acid and sarcosine in human serum, indicating the developed device has great potential in self-health management and clinical requirements.

Facile fabrication of hydrophilic covalent organic framework composites for highly selective enrichment of N-glycopeptides.

The development of facilely synthetic materials acts an essential role in glycoproteome analysis, especially for the highly efficient enrichment of N-linked glycopeptides. In this work, a facile and timesaving route was introduced in which COFTP-TAPT served as a carrier and poly (ethylenimine) (PEI) and carrageenan (Carr) were successively coated on the surface via electrostatic interaction. The resultant COFTP-TAPT@PEI@Carr showed remarkable performance in glycopeptide enrichment with high sensitivity (2 fmol µL-1), high selectivity (1:800, molar ratio of human serum IgG to BSA digests), large loading capacity (300 mg g-1), satisfactory recovery (102.4 ± 6.0%) and reusability (at least eight times). Due to the brilliant hydrophilicity and electrostatic interactions between COFTP-TAPT@PEI@Carr and positively charged glycopeptides, the prepared materials could be applied in the identification and analysis in the human plasma of healthy subjects and patients with nasopharyngeal carcinoma. As a result, 113 N-glycopeptides with 141 glycosylation sites corresponding to 59 proteins and 144 N-glycopeptides with 177 glycosylation sites corresponding to 67 proteins were enriched from 2 µL plasma trypsin digests of the control groups and patients with nasopharyngeal carcinoma, respectively. 22 glycopeptides were identified only from the normal controls and 53 glycopeptides were detected only from the other set. The results demonstrated that this hydrophilic material was promising on a large scale and further N-glycoproteome research.

An electrochemical aptasensor based on multi-walled carbon nanotubes loaded with PtCu nanoparticles as signal label for ultrasensitive detection of adenosine.

Adenosine, as an endogenous nucleoside modulator, plays an important role in heart rate regulation, neurotransmission, and control of the respiratory system and thus it is significantly important to realize its sensitive detection. Herein, a highly sensitive electrochemical aptasensor for adenosine detection was proposed by using multi-walled carbon nanotubes (MWCNTs) as support matrix loading PtCu nanoparticles (PtCu-MWCNTs) to amplify signal. On one hand, disposable screen-printing gold electrodes (SPGEs) were used as superb sensing base to ensure the stable connection of aptamers 1 (ssDNA1). On the other hand, the PtCu-MWCNTs complex was synthesized through a one-pot method, which not only can precisely control the proportion of metal mass in the product but also exhibited superior electrocatalytic activity towards H2O2. The recognition reactions were achieved by stepwise incubation of ssDNA1, ssDNA2-PtCu-MWCNTs (denoted as ssDNA2-label), and adenosine on the SPGEs. As a result, the constructed electrochemical aptasensor exhibited a wide linear range from 10 nM to 1.0 µM with a low detection limit of 1.0 nM (S/N = 3) for adenosine detection. The aptasensor also successfully realized the adenosine detection in human serum samples, which means that the proposed aptasensor holds a potential application in point-of-care detection.

Colon-specific delivery of methotrexate using hyaluronic acid modified pH-responsive nanocarrier for the therapy of colitis in mice.

Oral immunosuppressant methotrexate (MTX) is an effective method for the treatment of inflammatory bowel disease (IBD). To overcome the defects of clinical application of MTX, poly (lactic-co-glycolic acid) (PLGA), Eudragits® S100 (ES100), chitosan (CS) and hyaluronic acid (HA) were used to structure the MTX-loaded HA-CS/ES100/PLGA nanoparticles (MTX@hCEP). MTX@hCEP had a hydrodynamic particle size of approximately 202.5 nm, narrow size distribution, negative zeta potential (-18.7 mV), and smooth surface morphology. In vitro drug release experiments under simulated gastrointestinal conditions indicated that MTX@hCEP exhibited colonic pH-sensitive drug release properties. The cellular uptake capacity of hCEP nanoparticles was significantly enhanced in RAW 264.7 macrophages. Moreover, we further found that the MTX@hCEP also inhibited the proliferation and the secretion of pro-inflammatory cytokines in the LPS-stimulated macrophages. In vivo imaging results not only demonstrated that the accumulated in the colon of colitis mice, but also indicated the extended retention time of MTX in the colon. Additionally, MTX@hCEP alleviated inflammatory symptoms via decreasing the activities of myeloperoxidase and pro-inflammatory factors, promoting mucosal repair in vivo. Collectively, these results clearly demonstrated that MTX@hCEP with properties of colon-specific and macrophages targeting can be exploited as an efficient nanotherapeutic for IBD therapy.

Flexible electrochemical sensor with Fe/Co bimetallic oxides for sensitive analysis of glucose in human tears.

The construction of uniformly dispersed structure with abundant active sites is crucial for fast electron transport and advancing electrocatalytic reactions. Herein, FexCoyO4-rGO was prepared by depositing Fe and Co bimetallic oxides in-situ on reduced graphene oxide through a simple process combined hydrothermal reaction and calcination. Fe was elaborately introduced into the synthesis of metal oxides to alleviate the aggregation of cobalt oxides and obtain nanocomposites with homogeneously structured and abundant redox sites, and the bimetallic oxides nanomaterials had enhanced electrocatalysis under the synergistic effect. The flexible electrode prepared from FexCoyO4-rGO exhibited excellent detection performance for glucose with a detection limit down low to 0.07 µM and a sensitivity of 1510 µM cm-2 mA-1. The adoption of flexible substrates improved the wearability of the electrode and broadened its practicality for detecting biomarkers on the skin surface. The constructed sensor was successfully used in the dynamic analysis of glucose content in tears, and the results were highly consistent with the test outcome of a commercial test kit, demonstrating its application prospects in non-invasive epidermal diabetes mellitus diagnosis.

Development of a novel microfluidic biosensing platform integrating micropillar array electrode and acoustic microstreaming techniques.

Quantifying biomarkers at the early stage of the disease is challenging due to the low abundance of biomarkers in the sample and the lack of sensitive techniques. This article reports the development of a novel microfluidic electrochemical biosensing platform to address this challenge. The electrochemical sensing is achieved by utilizing a micropillar array electrode (µAE) coated with 3D bimetallic Pt-Pd nanotrees to enhance the sensitivity. A bubble-based acoustic microstreaming technique is integrated with the device to increase the contact of analyte molecules with the surface of electrodes to further enhance the electrochemical performance. The current density of Pt-Pd NTs/µAE with acoustic microstreaming is nearly 22 times that of the bare planar electrode in potassium ferrocyanide solution. The developed biosensor has demonstrated excellent sensing performance. For hydrogen peroxide detection, both the Pt-Pd NTs/µAE and acoustic microstreaming contribute to the sensitivity enhancement. The current density of the Pt-Pd NTs/µAE is approximatively 28 times that of the bare µAE. With acoustic microstreaming, this enhancement is further increased by nearly 1.6 times. The platform has a linear detection range of 5-1000 µM with a LOD of 1.8 µM toward hydrogen peroxide detection, while for sarcosine detection, the linear range is between 5 and 100 µM and LOD is 2.2 µM, respectively. Furthermore, the sarcosine biosensing shows a high sensitivity of 667 µA mM-1∙cm-2. Such a sensing platform has the potential as a portable device for high sensitivity detection of biomarkers.

Spatiotemporally controllable diphtherin transgene system and neoantigen immunotherapy.

Individualized immunotherapy has attracted great attention due to its high specificity, effectiveness, and safety. We used an exogenous antigen to label tumor cells with MHC I molecules, which allowed neoantigen-specific T cells to recognize and kill tumor cells. A neoantigen vaccine alone cannot achieve complete tumor clearance due to a tumor immunosuppressive microenvironment. The LightOn system was developed to effectively eliminate tumor cells through the spatiotemporally controllable expression of diphtheria toxin A fragment, leading to antigen release in the tumor region. These antigens stimulated and enhanced immunological function and thus, recruited neoantigen-specific T cells to infiltrate tumor tissue. Using the nanoparticle delivery system, neoantigens produced higher delivery efficiency to lymph nodes and improved tumor targeting ability for tumor cell labelling. Good tumor inhibition and prolonged survival were achieved, while eliciting a strong immune response. The combination of a spatiotemporally controllable transgene system with tumor neoantigen labeling has great potential for tumor immunotherapy.

XA pH-Responsive and Colitis-Targeted Nanoparticle Loaded with Shikonin for the Oral Treatment of Inflammatory Bowel Disease in Mice.

Epidemiology shows that more than 6.8 million people in the world are influenced by inflammatory bowel disease (IBD) each year. IBD is a refractory inflammatory disease, and the disease mainly affects the colon. Shikonin (SK) was originally extracted from traditional Chinese medicine "Zicao" (with an English name Lithospermum erythrorhizon) and found to inhibit inflammation, regulate immunity, and be involved in healing wounds. Herein, we used chitosan (CS), hyaluronic acid (HA), and pH-responsive polymer Eudragits S100 (ES100) to design SK-loaded ES100/HA/CS nanoparticles (SK@SAC) as an oral delivery system to treat the colitis mice. Particle size of SK@SAC was 190.3 nm and drug loading efficiency was 6.6%. SAC nanoparticles accumulated in RAW264.7 macrophages and exhibited colitis-targeted ability by increasing the local drug concentration as well as reducing nonspecific distribution after oral gavage. In TNBS-induced IBD mice, SK@SAC treatment had significant therapeutic effects, regulated of pro-inflammatory cytokines (TNF-α, IL-6, and IL-1ß) and anti-inflammatory cytokines (IL-10 and TGF-ß), and also inhibited COX-2 and iNOS activity. SK@SAC also increased tight junction protein ZO-1 and occludin to some extent. These promising results showed that this novel oral SK-loaded nanoparticle drug delivery system for targeted treatment provides a new strategy for the management of IBD.

A Ti3 C2 TX /Pt-Pd based amperometric biosensor for sensitive cancer biomarker detection.

The detection of cancer biomarkers is of great significance for the early screening of cancer. Detecting the content of sarcosine in blood or urine has been considered to provide a basis for the diagnosis of prostate cancer. However, it still lacks simple, high-precision and wide-ranging sarcosine detection methods. In this work, a Ti3 C2 TX /Pt-Pd nanocomposite with high stability and excellent electrochemical performance has been synthesized by a facile one-step alcohol reduction and then used on a glassy carbon electrode (GCE) with sarcosine oxidase (SOx ) to form a sarcosine biosensor (GCE/Ti3 C2 TX /Pt-Pd/SOx ). The prominent electrocatalytic activity and biocompatibility of Ti3 C2 TX /Pt-Pd enable the SOx to be highly active and sensitive to sarcosine. Under the optimized conditions, the prepared biosensor has a wide linear detection range to sarcosine from 1 to 1000 µM with a low limit of detection of 0.16 µM (S/N = 3) and a sensitivity of 84.1 µA/mM cm2 . Besides, the reliable response in serum samples shows its potential in the early diagnosis of prostate cancer. More importantly, the successful construction and application of the amperometric biosensor based on Ti3 C2 TX /Pt-Pd will provide a meaningful reference for detecting other cancer biomarkers.

Sandwich-type electrochemical aptasensor based on Au-modified conductive octahedral carbon architecture and snowflake-like PtCuNi for the sensitive detection of cardiac troponin I.

The cardiac troponin I (cTnI) detection is increasingly significant given its promising value in the clinical acute myocardial infarction diagnosis. Here a sensitive sandwich-type cTnI electrochemical aptasensor was developed by using zirconium-carbon loaded with Au (Au/Zr-C) as electrode-modified material and snowflake-like PtCuNi catalyst as label material. The Au/Zr-C was prepared from a carbonation process and a reduction step. The PtCuNi was synthesized by a one-pot hydrothermal reaction. On the one hand, due to its many merits of large effective area, rich pores, high degree of graphitization, the assistance of Au, the Au/Zr-C exhibited remarkable electronic conductivity but low catalytical capacity, thus improving the electrochemical property but lowing the background signal of electrode. On the other hand, because of its accessible active sites of the special snowflake-like structure and the synergy of three elements, the PtCuNi catalyst presented excellent catalytic activity and improved stability compared to binary alloy. The recognition reactions were achieved by stepwise incubation of aptamer 1, cTnI, and aptamer 2-PtCuNi (denoted as Apt2-label) on the Au/Zr-C-modified electrode. The electrocatalytic signals of the immobilized Apt2-label towards the H2O2 reduction were recorded in all tests for cTnI analysis. Consequently, this cTnI aptasensor exhibited excellent performance involving a wide linear range of 100 ng mL-1 to 0.01 pg mL-1 with a detection limit of 1.24 × 10-3 pg mL-1 (S/N = 3), good selectivity, satisfying reproducibility, outstanding stability, and good recovery.

Two-dimensional mesoporous nitrogen-rich carbon nanosheets loaded with CeO2 nanoclusters as nanozymes for the electrochemical detection of superoxide anions in HepG2 cells.

Two-dimensional (2D) porous carbon-based composite nanosheets loaded with metal oxide nanoclusters are expected to be promising electrocatalysts for high-performance electrochemical sensors. However, for this complicated composite material, strict reaction conditions and complex synthesis steps limit its general application in electrochemical detection. Here we present a facile method to fabricate 2D mesoporous nitrogen-rich carbon nanosheets loaded with CeO2 nanoclusters (2D-mNC@CeO2), for fabricating superoxide anions (O2•-) electrochemical sensor. The method is based on block copolymers self-assembly and the affinity of polydopamine to metal ions to obtain organic-inorganic hybrid, which can be directly converted into 2D-mNC@CeO2 through carbonization strategy without structural deterioration. Characterizations demonstrate that the 2D-mNC@CeO2 owned the 2D N-doped carbon structure with an interlinked hierarchical mesoporous and the uniformly dispersed CeO2 nanoclusters on the surface. Benefitted from the unique structure, the 2D-mNC@CeO2 shortens electron transfer distance, enhances mass transfer efficiency, exposes numerous active sites, and obtain a high Ce3+/Ce4+ ratio for improving electrocatalytic performance. The 2D-mNC@CeO2/SPCEs sensors for O2•- detection has a detection limit of 0.179 µM (S/N = 3) and sensitivity of 401.4 µA cm-2 mM-1. The sensors can be applied to capture electrochemical signals of O2•- released from HepG2 cells, demonstrating the application potential of the sensors to monitor O2•- in biological fields.

High-efficiency two-dimensional separation of natural products based on ß-cyclodextrin stationary phase working in both hydrophilic and reversed hydrophobic modes.

Natural products are rather complex samples containing a large number of compounds ranging from polar to nonpolar, small molecules to macromolecules, as well as numerous homologs/analogs with quite similar structures, which create great opportunities and treasures for discovery of bioactive drugs. For the purpose of better understanding the complex natural products and controlling their qualities, powerful analytical techniques for adequately separating chemical constituents and tracking potentially bioactive components are quite essential. Here, a design concept of bidirectional ß-cyclodextrin (ß-CD)-modified chromatographic stationary phase toward separation of bufadienolides extracted from toad is proposed. Bufadienolides could be divided into two classes: toad venom ligand (AAUBs) and toad toxin (AACBs) with remarkable differences in structures and polarity. The hydrophobic cavity of ß-CD can encapsulate the steroid part of AACBs while the hydroxyls exposed on the ß-CD surface have strong hydrophilic interactions with the arginine part of AACBs. Isothermal titration calorimetry and hydrogen nuclear magnetic resonance titration experiment further validate the rationality of this design. Furthermore, the ß-CD-based stationary phase can be used as a hydrophilic material to construct a HILIC × RPLC 2D separation mode for the separation of AACBs, also works as a reverse phase material to construct a RPLC × RPLC 2D separation mode for the separation of AAUBs with good orthogonality. This study will open an avenue and provide a novel insight for high-efficiency two-dimensional separation of natural products.

Light-switchable diphtherin transgene system combined with losartan for triple negtative breast cancer therapy based on nano drug delivery system.

Breast cancer is a common malignancy in women. The abnormally dense collagen network in breast cancer forms a therapeutic barrier that hinders the penetration and anti-tumor effect of drugs. To overcome this hurdle, we adopted a therapeutic strategy to treat breast cancer which combined a light-switchable transgene system and losartan. The light-switchable transgene system could regulate expression of the diphtheria toxin A fragment (DTA) gene with a high on/off ratio under blue light and had great potential for spatiotemporally controllable gene expression. We developed a nanoparticle drug delivery system to achieve tumor microenvironment-responsive and targeted delivery of DTA-encoded plasmids (pDTA) to tumor sites via dual targeting to cluster of differentiation-44 and αvß3 receptors. In vivo studies indicated that the combination of pDTA and losartan reduce the concentration of collagen type I from 5.9 to 1.9 µg/g and decreased the level of active transforming growth factor-ß by 75.0% in tumor tissues. Moreover, deeper tumor penetration was achieved, tumor growth was inhibited, and the survival rate was increased. Our combination strategy provides a novel and practical method for clinical treatment of breast cancer.

Nanostructure stable hydrophilic hierarchical porous metal-organic frameworks for highly efficient enrichment of glycopeptides.

Protein glycosylation plays a vital role in many physiological activities in organisms. Due to the low abundance of glycopeptides and the interference of numerous non-glycopeptides in biological samples, selective enrichment of glycopeptides is of great significance for their successful identification. Metal organic frameworks (MOFs) materials are appropriate for glycopeptides enrichment by virtue of their large specific surface area and outstanding hydrophilic properties. However, the instability of hydrophilic MOFs in acidic solutions have severely limited their applications. In this work, a rational facile strategy was established to synthesize a stable hydrophilic hierarchical porous MOF (denoted as HP-MOF-Arg@mSiO2). This new material improved the selectivity and sensitivity of enrichment for glycopeptides via modification of arginine groups. More importantly, the mesoporous silica layer was introduced to enhance the stability of MOFs in aqueous solution and achieve the size exclusion effect of large-size proteins in complex samples. Overall, owing to the unique hierarchical porous and the hydrophilic modification, the synthesized HP-MOF-Arg@mSiO2 materials showed excellent hydrophilicity and hydrolytic stability, resulting in outstanding specific separation capacity in glycopeptides enrichment. A total of 521 and 342 glycopeptides were respectively captured from 2 µL human serum digests and mouse testis tissue digests, revealing the potential of the materials in the study of glycoproteomics in complex biological samples.