Search

A compact and clamped CHA-control-HCR (CCH) circuitry system, specifically for amplified multi-microRNA imaging, is developed to precisely distinguish cancer cells from normal cells.

Objective. Mitochondrial dysfunction plays an important role in intervertebral disc degeneration (IDD). We aim to explore the pathways and key genes that cause mitochondrial dysfunction during IDD and to further reveal the pathogenesis of IDD based on bioinformatic analyses. Methods. Datasets GSE70362 and GSE124272 were downloaded from the Gene Expression Omnibus. Differentially expressed genes (DEGs) of mitochondrial dysfunction between IDD patients and healthy controls were screened by package limma package. Critical genes were identified by adopting gene ontology (GO), Kyoto encyclopedia of genes and genomes (KEGG) pathways, and protein-protein interaction (PPI) networks. We collected both degenerated and normal disc tissues obtained surgically, and we performed western blot and qPCR to verify the key DEGs identified in intervertebral disc tissues. Results. In total, 40 cases of IDD and 24 healthy controls were included. We identified 152 DEGs, including 67 upregulated genes and 85 downregulated genes. Four genes related to mitochondrial dysfunction (SOX9, FLVCR1, NR5A1 and UCHL1) were screened out. Of them, SOX9, FLVCR1, and UCHL1 were down-regulated in peripheral blood and intervertebral disc tissues of IDD patients, while NR5A1 was up-regulated. The analysis of immune infiltration showed the concentrations of mast cells activated were significantly the highest in IDD patients. Compared with the control group, the level of T cells CD4 memory resting was the lowest in the patients. In addition, 24 cases of IDD tissues and 12 cases of normal disc tissues were obtained to verify the results of bioinformatics analysis. Both western blot and qPCR results were consistent with the results of bioinformatics analysis. Conclusion. We identified four genes (SOX9, FLVCR1, NR5A1 and UCHL1) associated with mitochondrial dysfunction that play an important role in the progress of disc degeneration. The identification of these differential genes may provide new insights for the diagnosis and treatment of IDD.

S1: RT-PCR confirms expression of E6 and E7 in TC1 S2: CD4+ T cell responses after E6E7 vaccination S3: E6E7 vaccination induces T-cell degranulation Tumor free S4: Tumor volume curves in E6E7 vaccinated mice S5: TCM frequency in mice after TC1 tumor clearance S6: PCA analysis of TC1 tumors S7: Images of MG1-GFP infected HPV+ primary tumor

The viral-transforming proteins E6 and E7 make human papillomavirus–positive (HPV+) malignancies an attractive target for cancer immunotherapy. However, therapeutic vaccination exerts limited efficacy in the setting of advanced disease. We designed a strategy to induce substantial specific immune responses against multiple epitopes of E6 and E7 proteins based on an attenuated transgene from HPV serotypes 16 and 18 that is incorporated into MG1-Maraba virotherapy (MG1-E6E7). Mutations introduced to the transgene abrogate the ability of E6 and E7 to perturb p53 and retinoblastoma, respectively, while maintaining the ability to invoke tumor-specific, multifunctional CD8+ T-cell responses. Boosting with MG1-E6E7 significantly increased the magnitude of T-cell responses compared with mice treated with a priming vaccine alone (greater than 50 × 106 E7-specific CD8+ T cells per mouse was observed, representing a 39-fold mean increase in boosted animals). MG1-E6E7 vaccination in the HPV+ murine model TC1 clears large tumors in a CD8+-dependent manner and results in durable immunologic memory. MG1-Maraba can acutely alter the tumor microenvironment in vivo and exploit molecular hallmarks of HPV+ cancer, as demonstrated by marked infection of HPV+ patient tumor biopsies and is, therefore, ideally suited as an oncolytic treatment against clinical HPV+ cancer. This approach has the potential to be directly translatable to human clinical oncology to tackle a variety of HPV-associated neoplasms that cause significant morbidity and mortality globally. Cancer Immunol Res; 5(10); 847–59. ©2017 AACR.

An on-site activated aptasensing platform is constructed for highly reliable ATP monitoring in cancer cells both in vitro and in vivo by combining tumor-specific activatable MnO2 with the stimuli-responsive cascade DNA amplifier.

Constructing artificial domino nanoarchitectures, especially dynamic DNA circuits associated with the actuation of biological functions inside live cells, represents a versatile and powerful strategy to regulate the behaviors and fate of various living entities. However, the stepwise operation of conventional DNA circuits always relies on freely diffusing reactants, which substantially slows down their operation rate and efficiency. Herein, a self-adaptive localized catalytic circuit (LCC) is developed to execute the self-sustained bioorthogonal assembly of DNA nanosponges within a crowded intracellular environment. The LCC-generated DNA scaffolds are utilized as versatile templates for realizing the proximity confinement of LCC reactants. Single-molecule-detecting fluorescence correlation spectroscopy (FCS) is used to explore the reaction acceleration of the catalytic circuit. This self-adaptive DNA circuit facilitates the bioorthogonal assembly of highly branched DNA networks for robust and accurate monitoring of miRNA targets. Based on its intriguing and modular design, the LCC system provides a pivotal molecular toolbox for future applications in early disease diagnosis.

Oxidative stress modulation is effective to kill cancer. While considerable effort has focused on the promotion of reactive oxygen species (ROS) levels, an additional crucial aspect of ROS regulati...

The direct visualization of vaccine fate is important to investigate its immunoactivation process to elucidate the detailed molecular reaction process at single‐molecular level. Yet, visualization of the spatiotemporal trafficking of vaccines remains poorly explored. Here, we show that quantum dot (QD) nanomaterials allow for monitoring vaccine dynamics and for amplified immune response. Synthetic QDs enable efficient conjugation of antigen and adjuvants to target tissues and cells, and non‐invasive imaging the trafficking dynamics to lymph nodes and cellular compartments. The nanoparticle vaccine elicits potent immune responses and anti‐tumor efficacy alone or in combination with programmed cell death protein 1 blockade. The synthetic QDs showed high fluorescence quantum yield and superior photostability, and the reliable and long‐term spatiotemporal tracking of vaccine dynamics was realized for the first time by using the synthetic QDs, providing a powerful strategy for studying immune response and evaluating vaccine efficacy., Visualization of spatiotemporal trafficking of vaccines is achieved with synthetic QDs for potent immune activation.

The systemic therapeutic utilisation of RNA interference (RNAi) is limited by the non-specific off-target effects, which can have severe adverse impacts in clinical applications. The accurate use of RNAi requires tumour-specific on-demand conditional activation to eliminate the off-target effects of RNAi, for which conventional RNAi systems cannot be used. Herein, a tumourous biomarker-activated RNAi platform is achieved through the careful design of RNAi prodrugs in extracellular vesicles (EVs) with cancer-specific recognition/activation features. These RNAi prodrugs are assembled by splitting and reconstituting the principal siRNAs into a hybridisation chain reaction (HCR) amplification machine. EVs facilitate the specific and efficient internalisation of RNAi prodrugs into target tumour cells, where endogenous microRNAs (miRNAs) promote immediate and autonomous HCR-amplified RNAi activation to simultaneously silence multiantenna hypoxia-related genes. With multiple guaranteed cancer recognition and synergistic therapy features, the miRNA-initiated HCR-promoted RNAi cascade holds great promise for personalised theranostics that enable reliable diagnosis and programmable on-demand therapy., The therapeutic application of small interfering RNA (siRNA) is challenging due to its non-specific targeting and delivery issues. Here, the authors report an endogenous micro-RNA guided and hybridisation chain reaction-promoted siRNA delivery system encapsulated in tumour-derived extracellular vesicles, with cancer-specific activation, and achieve silencing of hypoxia-related genes.

Nucleic acid therapeutics has reached clinical utility through modulating gene expression. As a potential oligonucleotide drug, DNAzyme has RNA-cleaving activity for gene silencing, but faces challenges due to the lack of a safe and effective delivery vehicle and low

Lead zirconate titanate (PZT) films have been deposited on Ti–Pt/Ti/SiO2/Si(100) alloy substrates by sol-gel process. The purpose of the present study was to examine the Ti content in the Ti–Pt alloy layers-dependent preferred orientation and electrical behaviors of PZT films. X-ray diffraction (XRD) shows that Ti–Pt alloy electrodes induces texture transition from (100) to (111), and the maximum peak of (100) and minimum peak of (111) are obtained in the sample on Ti–Pt alloy layer with a sputtering power of 12.5 W/150 W. The film deposited on Ti–Pt alloy layer with a sputtering power of 12.5 W/150 W exhibits a reduced oxygen vacancies concentration, which results in a maximum e of 1224.25, an enhanced 2Pr of 40.5 μC/cm2, and a relative decreased 2Ec of 108 kV/cm. Therefore, it can be concluded that the Ti–Pt alloy layers with an appropriate Ti content play a very critical role for the preferred orientation, microstructure, and improved electrical properties of the PZT films.

The extreme complexity and heterogeneity of fatal tumors requires the development of combination phototherapy considering the limited therapeutic efficiency of conventional monomodal photodynamic therapy (PDT) or photothermal therapy (PTT). However, tumor-specific drug administration and the accompanying hypoxia-restrained PDT present the main obstacles for executing an efficient combination phototherapy. Developing a highly biocompatible, tumor-specific, near infrared absorbing, and oxygen (O2)-evolving multifunctional nanoplatform is thus crucial for an effective PDT-based combination therapy. In this contribution, a multifunctional ZIF-8-gated polydopamine nanoparticle (PDA) carrier was synthesized for simultaneously delivering a photosensitizer and a catalase (CAT) into tumor cells, thus realizing a cooperatively enhanced combination photodynamic and photothermal therapy, as systematically demonstrated in vitro and in vivo. The ZIF-8 gatekeeper facilitates the simultaneous and effective delivery of these functional payloads, and the subsequent tumor acidic pH-stimulated drug release. This leads to a significant improvement of combination efficacy by ameliorating tumor hypoxic conditions since the CAT-mediated self-sufficient O2 generation could substantially promote an efficient PDT operation. In addition, this nanoplatform can effectively convert near infrared photoradiation into heat, resulting in thermally induced elimination of cancerous cells. As an intelligent multi-mode therapeutic nanosystem, this inorganic/organic hybrid nanosystem showed great potential for accurate cancer diagnosis and immediate therapy.

Powerful information processing and ubiquitous computing are crucial for all machines and living organisms. The Watson-Crick base-pairing principle endows DNA with excellent recognition and assembly abilities, which facilitates the design of DNA computers for achieving intelligent systems. However, current DNA computational systems are always constrained by poor integration efficiency, complicated device structures or limited computational functions. Here, we show a DNA arithmetic logic unit (ALU) consisting of elemental DNA logic gates using polymerase-mediated strand displacement. The use of an enzyme resulted in highly efficient logic gates suitable for multiple and cascaded computation. Based on our basic single-rail DNA configuration, additional combined logic gates (e.g., a full adder and a 4:1 multiplexer) have been constructed. Finally, we integrate the gates and assemble the crucial ALU. Our strategy provides a facile strategy for assembling a large-scale complex DNA computer system, highlighting the great potential for programming the molecular behaviors of complicated biosystems., Current DNA computational systems are constrained by integration efficiency, device structures and limited functions. Here the authors design a DNA arithmetic logic unit that uses polymerase-mediated strand displacement.

Summary An elaborately designed multifunctional nanoplatform with intelligent and inherent tumor-specific theranostic capacity is urgently desired for highly efficient cancer treatments. Herein, we report a class of compact MnO2-laden black phosphorus (BPN/MnO2) nanostructure for delivering drugs with smart tumor-specific diagnosis and exquisite self-driven therapeutic features. The auxiliary MnO2 nanosheets promote the photodynamic therapy (PDT) of black phosphorus nanosheets (BPNs) by 3.8-fold through their inherent remarkable hypoxia amelioration. Moreover, BPN/MnO2 shows an obvious 37% photothermal therapy (PTT) enhancement relative to BPNs. Significantly, the multiple-tumor-microenvironment-responsive features of MnO2 contribute to the maximized drug administration on the tumor (≥4-fold drug release of the single acid stimuli), thus reducing the potential side effect of chemotherapy on normal tissues. Besides the spatiotemporally programmable administration of drugs, BPN/MnO2 also provides intelligent MRI and thermal imaging guidance for synergistically enhancing PDT, PTT, and chemotherapy, thus offering an intelligent versatile toolbox for multifunctional nanotheranostic applications.

The accurate intracellular imaging of metal ions requires an exquisite site-specific activation of metal-ion sensors, for which the pervasive epigenetic regulation strategy can serve as an ideal alternative thanks to its orthogonal control feature and endogenous cell/tissue-specific expression pattern. Herein, a simple yet versatile demethylation strategy was proposed for on-site repairing-to-activating the metal-ion-targeting DNAzyme and for achieving the accurate site-specific imaging of metal ions in live cells. This endogenous epigenetic demethylation-regulating DNAzyme system was prepared by modifying the DNAzyme with an m6A methylation group that incapacitates the DNAzyme probe, thus eliminating possible off-site signal leakage, while the cell-specific demethylase-mediated removal of methylation modification could efficiently restore the initial catalytic DNAzyme for sensing metal ions, thus allowing a high-contrast bioimaging in live cells. This epigenetic repair-to-activate DNAzyme strategy may facilitate the robust visualization of disease-specific biomarkers for in-depth exploration of their biological functions., A simple yet versatile demethylation strategy is proposed for an on-site repairing-to-activating metal-ion-targeting DNAzyme and for achieving the highly reliable site-specific imaging of metal ions in live cells.

Catalytic DNA circuits represent a versatile toolbox for tracking intracellular biomarkers yet are constrained with low anti-interference capacity originating from their severe off-site activation. Herein, by introducing an unprecedented endogenous DNA repairing enzyme-powered pre-selection strategy, we develop a sequential and specific on-site activated catalytic DNA circuit for achieving the cancer cell-selective imaging of microRNA with high anti-interference capacity. Initially, the circuitry reactant is firmly caged by an elongated stabilizing duplex segment with a recognition/cleavage site of a cell-specific DNA repairing enzyme, which can prevent undesired signal leakage prior to its exposure to target cells. Then, the intrinsic DNA repairing enzyme of target cells can liberate the DNA probe for efficient intracellular microRNA imaging via the multiply guaranteed molecular recognition/activation procedures. This bioorthogonal regulated DNA circuit presents a modular and programmable amplification strategy for highly reliable assays of intracellular biomarkers, and provides a pivotal molecular toolbox for living systems., An on-site bioorthogonal regulated DNA circuit was developed by introducing an endogenous DNA repairing enzyme-mediated sequential activation strategy to achieve cancer cell-selective microRNA imaging with high anti-interference ability.

SummaryThe emerging SARS-CoV-2 variants of concern (VOC) increasingly threaten the effectiveness of current first-generation COVID-19 vaccines that are administered intramuscularly and are designed to only target the spike protein. There is thus a pressing need to develop next-generation vaccine strategies to provide more broad and long-lasting protection. By using adenoviral vectors (Ad) of human and chimpanzee origin, we developed Ad-vectored trivalent COVID-19 vaccines expressing Spike-1, Nucleocapsid and RdRp antigens and evaluated them following single-dose intramuscular or intranasal immunization in murine models. We show that respiratory mucosal immunization, particularly with chimpanzee Ad-vectored vaccine, is superior to intramuscular immunization in induction of the three-arm immunity, consisting of local and systemic antibody responses, mucosal tissue-resident memory T cells, and mucosal trained innate immunity. We further show that single-dose intranasal immunization provides robust protection against not only the ancestral strain of SARS-CoV-2, but also two emerging VOC, B.1.1.7 and B.1.351. Our findings indicate that single-dose respiratory mucosal delivery of an Ad-vectored multivalent vaccine represents an effective next-generation COVID-19 vaccine strategy against current and future VOC. This strategy has great potential to be used not only to boost first-generation vaccine-induced immunity but also to expand the breadth of protective T cell immunity at the respiratory mucosa.

The emerging SARS-CoV-2 variants of concern (VOCs) threaten the effectiveness of current COVID-19 vaccines administered intramuscularly and designed to only target the spike protein. There is a pressing need to develop next-generation vaccine strategies for broader and long-lasting protection. Using adenoviral vectors (Ad) of human and chimpanzee origin, we evaluated Ad-vectored trivalent COVID-19 vaccines expressing spike-1, nucleocapsid, and RdRp antigens in murine models. We show that single-dose intranasal immunization, particularly with chimpanzee Ad-vectored vaccine, is superior to intramuscular immunization in induction of the tripartite protective immunity consisting of local and systemic antibody responses, mucosal tissue-resident memory T cells and mucosal trained innate immunity. We further show that intranasal immunization provides protection against both the ancestral SARS-CoV-2 and two VOC, B.1.1.7 and B.1.351. Our findings indicate that respiratory mucosal delivery of Ad-vectored multivalent vaccine represents an effective next-generation COVID-19 vaccine strategy to induce all-around mucosal immunity against current and future VOC.

DNAzyme-based chemiluminescence assay exhibits excellent performance in bioanalysis but their operation in acid conditions remains challengeable. Herein, we constructed an acid-improved DNAzyme-based isothermal enzyme-free concatenated DNA circuit with significantly reduced background and simultaneously improved signal-to-noise ratio for miRNA detection. The chemiluminescence miRNA assay is composed of catalyzed hairpin assembly (CHA), hybridization chain reaction (HCR), and hemin/G-quadruplex DNAzyme units. The analyte initiates the self-assembly of CHA hairpins into numerous dsDNA, which triggers the subsequent autonomous cross-opening of HCR hairpins to generate long nanowires consisting of the hemin/G-quadruplex DNAzyme. The DNAzyme catalyzes the oxidation of luminol by hydrogen peroxide for the cascaded amplified chemiluminescence signal. The acid-improved property was demonstrated to be closely associated with the low catalytic activity of aggregated hemin under acidic conditions and the remained multiple amplified signal through concatenated DNA circuit. The general DNA circuit exhibited high sensitivity for miRNA-21 detection and chemiluminescence imaging under acidic conditions with a recognition hairpin. The acid-improved DNAzyme-based concatenated DNA circuit is promising to expand the application of chemiluminescence assay and provide a valuable strategy for early diagnosis and prognosis of cancer.

Hengshuo Zhang, Lu Chen, Ziyu Wang, Fuan Wang, Yu Shan, Linzeng Qi, Yunzhen Chen Department of Spine Surgery, Qilu Hospital, Cheeloo College of Medicine, Shandong University, Jinan, People’s Republic of ChinaCorrespondence: Yunzhen Chen Email qilucyz@yeah.netObjective: To examine the association between single nucleotide polymorphisms (SNPs) rs2228570, rs731236, rs7975232, and rs1544410 and lumbar disc degeneration (LDD) predisposition.Methods: A search strategy was carried out, and the data were extracted after being chosen by the inclusion and exclusion criteria. Pooled odds ratios and 95% confidence intervals were calculated to assess the association between the aforementioned SNPs and LDD under allelic, dominant, recessive, heterozygous, and homozygous genetic models. In addition, a case–control study involving 46 LDD cases and 45 controls was also performed in the analysis to verify the result.Results: A total of 17 studies were included in this meta-analysis. The pooled results did not show any association between vitamin D receptor (VDR) gene polymorphisms and LDD. But, interestingly, in subgroup analysis, the rs2228570 polymorphism was associated with LDD under the allelic (OR = 0.70, 95% CI = 0.56– 0.87, p = 0.002), recessive (OR = 0.60, 95% CI = 0.43– 0.84, p = 0.003), and homozygous (OR = 0.47, 95% CI= 0.28– 0.79, p = 0.004) genetic models in the Asian population. SNPs rs731236 and rs7975232 still did not show any obvious association. We obtained a similar result from the case–control study: rs2228570 had an obvious relationship with LDD under allelic and homozygous genetic models. At the same time, we found that rs2228570 was also associated with the degree of low back pain (visual analogue scale, VAS score) in LDD population.Conclusion: SNP rs2228570 was significantly associated with LDD predisposition and the degree of low back pain in the Asian population.Keywords: vitamin D receptor, single nucleotide polymorphism, lumbar disc degeneration, visual analogue scale, meta-analysis, case-control study

Pb1−x(LaxZryTi1−y)O3 (PLZT) (x = 0%, 2%, 4%; y = 0.45, 0.52, 0.60) thin films were fabricated by sol–gel process to investigate the effects of La-doped and Zr/Ti ratio on crystalline orientation, microstructure, and electrical properties of lead zirconate titanate films. The results of X-ray diffraction analysis showed that the orientation of the films with various La concentration change with the Zr/Ti ratio. The Zr-rich composition (y = 0.60) with the fixed 2% La doping concentration has the most (100) preferential orientation. Scanning electron microscope analysis showed that the films exhibit dense perovskite structure when the doping concentration was less than 4%. The maximum dielectric constant (1364.65 at 100 Hz), optimized ferroelectric properties, and a low leakage current density of 8.63 × 10−8 A cm−2 were obtained for 2% La-doped film with a Zr/Ti ratio of 60/40. The lowest value of coercive field (Ec) was generated in the film fabricated with a Zr/Ti ratio of 52/48 when the La doping concentration was 2%. Fatigue resistance was also improved with 2% La dopant for the films processed by a Zr/Ti ratio of 52/48 and 60/40.

Biomolecular self-assembly circuits have been well developed for high-performance biosensing and bioengineering applications. Here we designed an isothermal concatenated nucleic acid amplification system which is composed of a lead-in catalyzed hairpin assembly (CHA), intermediate hybridization chain reaction (HCR) and ultimate DNAzyme amplifier units. The analyte initiates the self-assembly of hairpin reactants into dsDNA products in CHA, which generates numerous trigger sequences for activating the subsequent HCR-assembled long tandem DNAzyme nanowires. The as-acquired DNAzyme catalyzed the successive cleavage of its substrates, leading to an amplified fluorescence readout. The sophisticated design of our CHA-HCR-DNAzyme scheme was systematically investigated in vitro and showed dramatically enhanced detection performance. As a general sensing strategy, this CHA-HCR-DNAzyme method enables the amplified analysis of miRNA and its accurate intracellular imaging in living cells, originating from their synergistic signal amplifications. This method shows great potential for analyzing trace amounts of biomarkers in various clinical research studies.

A reconfigurable hybridization-based chain reaction was introduced to assemble enzyme-free DNA logic gates and advanced logic circuits for analyzing multiple endogenous miRNA expressions and discriminating different living cells., Dynamic nucleic acid-based biocircuits have spurred substantial research efforts for diagnosis or biomedical applications at the molecular level; nevertheless, it still remains a challenge to design programmable molecular circuit devices for autonomous and accurate diagnosis of low abundance biomolecules in a complex intracellular environment. Herein, a reconfigurable hybridization-based chain reaction is introduced to assemble modular biocomputing circuits that include a general sensing module and a versatile processing module. By modular sensing module design, we realized multiple endogenous miRNA-initiated biocomputing operations, including binary logic gates (OR, AND, INHIBIT and XOR), and more advanced concatenated logic circuits (XOR-AND, XOR-INHIBIT, and XOR-OR) in different living cells. The sensing module transduces the primary miRNA sensing event into an intermediate trigger for activating the processing module that further transduces the specific analyte recognition pattern into an amplified fluorescence readout. Based on an appropriate selection of multiple miRNA analytes, various miRNA expression patterns could be utilized for sensitive and selective cell discriminations. The inherent synergistically accelerated recognition and hybridization features of our biocomputing systems contribute to the amplified detection of multiplex endogenous miRNAs in living cells, thus providing an efficient toolbox for more accurate diagnosis and programmable therapeutics.

Pb1.2−xCexZr0.52Ti0.48O3 (PCZT, x = 0%, 0.1%, 0.5%, 1%, 2% and 3%) thin films with the thickness of about 1 µm were fabricated by sol–gel process and traditional annealing process on Pt/Ti/SiO2/Si substrates to investigate the effect of cerium doping on crystalline orientation, microstructure and electric properties of the samples. (100)-oriented Pb1.2−xCexZr0.52Ti0.48O3 films were obtained for all x values. The results of Scanning electron microscopy (SEM) revealed that the 0%, 0.1%, 0.5%, and 1% cerium doped Pb1.2−xCexZr0.52Ti0.48O3 films have a dense columnar perovskite structure. The maximum dielectric constant and remnant polarization were obtained for 0.1% Ce-doped film.

Extracellular vesicles (EVs) have been emerged as versatile drug delivery vehicles due to their outstanding biocompatibility and long-term circulation, yet are constrained with low targeting property and inefficient loading capacity from post-synthetic passive EVs encapsulation. Herein, we report a simple and feasible in situ biosynthetic approach to encapsulate tumor-targeting folate (FA)-modified EVs with intracellularly produced protoporphyrin X (PpIX) and doxorubicin (DOX). As compared with the traditional directly drug-incubated or drug-electroporated EVs, these biosynthesized EVs revealed high drug-loading efficiency with minimized structural and functional perturbations. Our multifunctional EVs revealed the enhanced accumulation and penetration into deep tumor parenchyma, as well as the strengthened immune response to ablate orthotopic and metastatic tumors, thus realizing the more reliable photochemotherapy. As an intelligent multi-mode therapeutic system, our biosynthetic EVs could be engineered with more therapeutic agents and show great promise for biomedicine applications.

The platelet isoform of phosphofructokinase (PFKP) is a rate‑limiting enzyme involved in glycolysis that serves an important role in various types of cancer. The aim of the present study was to explore the specific regulatory relationship between PFKP and non‑small cell lung cancer (NSCLC) progression. PFKP expression in NSCLC tissues and corresponding adjacent tissues was detected using reverse transcription‑quantitative polymerase chain reaction (RT‑qPCR) and immunohistochemical analysis. PFKP expression in human bronchial epithelial cells (16HBE) and NSCLC cells (H1299, H23 and A549) was also detected using RT‑qPCR. Cell proliferation was detected by Cell Counting Kit‑8 and colony formation assays. Transwell invasion and wound healing assays, and flow cytometry were used to detect cell invasion, migration and apoptosis, respectively. The expression levels of glycolysis‑associated enzymes (hexokinase‑2, lactate dehydrogenase A and glucose transporter‑1), epithelial‑mesenchymal transition‑related proteins (N‑cadherin, vimentin and E‑cadherin) and apoptosis‑related proteins (caspase‑3 and B‑cell lymphoma‑2) were detected by western blotting. Glucose uptake, lactate production and the adenosine trisphosphate/adenosine diphosphate ratio were measured using the corresponding kits. The results of the present study demonstrated that PFKP expression was upregulated in NSCLC tissues and cells, and PFKP expression was related to lymph node metastasis and histological grade. In addition, overexpression of PFKP inhibited cell apoptosis, and promoted proliferation, migration, invasion and glycolysis of H1299 cells, whereas knockdown of PFKP had the opposite effects. In conclusion, PFKP inhibited cell apoptosis, and promoted proliferation, migration, invasion and glycolysis of NSCLC cells; these findings may lay the foundation for novel treatments of NSCLC.

To investigate the effects of extracts from Chuanwu (Aconitum Carmichaelii) and Banxia (Rhizoma Pinelliae) on the excisional wound healing in a rat's model.Rats were performed a surgical lesion with a 2.0 cm resected tissue in the dorsal fascia. Following, animals were randomly divided into model group, YNB group and APE group those were respectively treated with saline, 1 mg/mL of Yunnan Baiyao and 1 mg/mL of Chuanwu (Aconitum Carmichaelii) and Banxia (Rhizoma Pinelliae) extracts. Wound contractions in days 0, 3, 7, 14 and 21 were calculated by an image analyzer. Histological analysis was analyzed using hematoxilin and eosin. Levels of tumor necrosis factor α (TNF-α), interleukin-2 (IL-2), IL-4, IL-10, transforming growth factor-β1 (TGF-β1) and basic fibroblast growth factor (bFGF) were determined by real-time quantitative PCR.Compared with that of YNB group and APE gtoup, the skin of rats showed poor re-modeling and re-epithelization characterized by a significant decrease of neovascularization, epithelialization and fibroblast in the model group. In the APE group, levels of TNF-α and IL-2 were significantly down-regulated and IL-4 and IL-10 significantly up-regulated in contrasted with that of model group. In addition, levels of TGF-β1 and bFGF in the APE group were significantly induced compared with that of model group.The results suggest that the extracts from Chuanwu (Aconitum Carmichaelii) and Banxia (Rhizoma Pinelliae) promote wound healing in the rats, which is associated with enhancing anti-inflammatory ability and inducing fibroblast formation.

The extreme complexity and heterogeneity of fatal tumors requires the development of combination phototherapy considering the limited therapeutic efficiency of conventional monomodal photodynamic therapy (PDT) or photothermal therapy (PTT)., The extreme complexity and heterogeneity of fatal tumors requires the development of combination phototherapy considering the limited therapeutic efficiency of conventional monomodal photodynamic therapy (PDT) or photothermal therapy (PTT). However, tumor-specific drug administration and the accompanying hypoxia-restrained PDT present the main obstacles for executing an efficient combination phototherapy. Developing a highly biocompatible, tumor-specific, near infrared absorbing, and oxygen (O2)-evolving multifunctional nanoplatform is thus crucial for an effective PDT-based combination therapy. In this contribution, a multifunctional ZIF-8-gated polydopamine nanoparticle (PDA) carrier was synthesized for simultaneously delivering a photosensitizer and a catalase (CAT) into tumor cells, thus realizing a cooperatively enhanced combination photodynamic and photothermal therapy, as systematically demonstrated in vitro and in vivo. The ZIF-8 gatekeeper facilitates the simultaneous and effective delivery of these functional payloads, and the subsequent tumor acidic pH-stimulated drug release. This leads to a significant improvement of combination efficacy by ameliorating tumor hypoxic conditions since the CAT-mediated self-sufficient O2 generation could substantially promote an efficient PDT operation. In addition, this nanoplatform can effectively convert near infrared photoradiation into heat, resulting in thermally induced elimination of cancerous cells. As an intelligent multi-mode therapeutic nanosystem, this inorganic/organic hybrid nanosystem showed great potential for accurate cancer diagnosis and immediate therapy.