[Virulence genes and antimicrobial resistance of Bacillus cereus in prepackaged pastries sold in Taizhou from 2020 to 2022].

OBJECTIVE: To investigate the virulence genes and antimicrobial resistance of Bacillus cereus from the pre-packaged pastries in Taizhou city. METHODS: 500 pre-packaged patries were collected in taizhou city market. 97 Bacillus cereus strains were detected from them by GB 4789.14-2014 method and identified with 4 houseking genes, then 13 virulence genes were detected by polymerase chain reaction(PCR)method and the antimicrobial resistance of Bacillus cereus to 19 antibiotics was detected by paper diffusion method. RESULTS: The result showed that the contamination rate of Bacillus cereus was 19.4% in 500 pre-packaged pastries. The detection rate of four housekeeping genes groEL, gyr B, rpoB and Vrr were 100%, 94.8%, 97.9% and 96.9%, respectively, and 89.7% at the same time. The virulence gene test result showed that the detection rate of nheABC, entFM, bceT, cytK and hblABCD were 91.8%, 88.7%, 61.9%, 51.6% and 25.8%, emetic virulence genes had the lowest detection rate, ces and EMl were 4.1%, cer was 5.2%. 97 Bacillus cereus strains show different degrees of drug resistance to 14 antimicrobials, the resistance rates to penicillin, ampicillin, cefotaxime and cotrimoxazole were higher than 95%, but they were completely sensitive to streptomycin, vancomycin and chloramphenicol. CONCLUSION: There is a risk of contamination by diarrhea-type Bacillus cereus and vomiting-type Bacillus cereus in prepackaged pastries in Taizhou. The isolated and identified Bacillus cereus has multiple-drug resistance.

Ultrasound-Chargeable Persistent Luminescence Nanoparticles to Generate Self-Propelled Motion and Photothermal/NO Therapy for Synergistic Tumor Treatment.

Cancer phototherapy indicates advantages in ease of manipulation, negligible drug resistance, and spatiotemporal control but is confronted with challenges in tumor cell accessibility and intermittent light excitation. Herein, we propose a strategy with persistent luminescence (PL)-excited photothermal therapy (PTT), concurrent thermophoresis-propelled motion, and PL-triggered NO release, where PL emission is chargeable by ultrasonication for readily applicable to deep tumors. Mechanoluminescent (ML) nanodots of SrAl2O4:Eu2+ (SAOE) and PL nanodots of ZnGa2O4:Cr3+ (ZGC) were deposited on mesoporous silicates to obtain mSZ nanoparticles (NPs), followed by partially coating with polydopamine (PDA) caps and loading NO donors to prepare Janus mSZ@PDA-NO NPs. The ML emission bands of SAOE nanodots overlap with the excitation band of ZGC, and the persistent near-infrared (NIR) emission could be repeatedly activated by ultrasonication. The PL emission acts as an internal NIR source to produce a thermophoretic force and NO gas propellers to drive the motion of Janus NPs. Compared with the commonly used intermittent NIR illumination at both 660 and 808 nm, the persistent motion of ultrasound-activated NPs enhances cellular uptake and long-lasting PTT and intracellular NO levels to combat tumor cells without the use of any chemotherapeutic drugs. The ultrasound-activated persistent motion promotes intratumoral accumulation and tumor distribution of PTT/NO therapeutics and exhibits significantly higher tumor growth inhibition, longer animal survival, and larger intratumoral NO levels than those who experience external NIR illumination. Thus, this study demonstrates a strategy to activate PL emissions and construct PL-excited nanomotors for phototherapy in deep tissues.

Pyroelectric Janus nanomotors to promote cell internalization and synergistic tumor therapy.

The tumor diffusion and cell internalization are the major obstacles to improving delivery efficacy of therapeutic agents. External electric fields have shown strong effect on the cell membrane polarization and fluidity, but usually need complicated power management circuits. Herein, in situ generation of microelectric field on nanoparticles (NPs) is proposed to overcome these delivery barriers. Janus tBT@PDA-CPT NPs were developed through partially coating of polydopamine (PDA) caps on pyroelectric tetragonal BaTiO3 (tBT) NPs and then camptothecin (CPT) conjugation via disulfide linkages. For comparison, cBT@PDA-CPT NPs were prepared from non-pyroelectric cubic BaTiO3 (cBT) as control. Near-infrared (NIR) illumination on PDA caps of the Janus NPs produces asymmetric thermophoretic force to drive NP motion for tumor accumulation, deep tissue penetration and effective cell interaction. Photothermally created temperature variations on tBT NPs build pyroelectric potentials to selectively change the membrane potential of tumor cells other than normal cells and exhibit a dominated role in enhancing tumor cell internalization and cytotoxicity. The combination index analysis confirms the synergistic effect of pyroelectric dynamic therapy (PEDT), chemotherapy and photothermal therapy (PTT), leading to full inhibition of tumor growth and noticeable extension of animal survival at significant lower CPT doses. The mild PTT/PEDT, the reduced CPT dose and the selective toxicity to tumor cells have achieved favorable treatment safety after tBT@PDA-CPT/NIR treatment. Therefore, in response to the differences in membrane potentials and glutathione levels between tumor and normal cells, we have demonstrated a concise design to achieve thermophoresis-driven motion, pyroelectric potential-enhanced cell internalization and PTT/PEDT/chemotherapy-synergized antitumor treatment.

Surface Wettability-Switchable Janus Fiber Fragments Stabilize Pickering Emulsions for Effective Oil/Water Separation.

Pickering emulsions indicate stronger resistance against droplet coalescence than the surfactant-stabilized emulsions. To resemble the surfactant amphiphilicity, Janus fiber fragments (JFs) were herein prepared through side-by-side electrospinning of poly(styrene-maleic anhydride) (PSMA) derivatives and cryosection of the aligned fibers, followed by conjugation of hydrophobic cetylamine (C16) and hydrophilic poly(N-isopropylacrylamide) (PNIPAm) ligands on the separate sides. Orthogonal analysis table L25(56) was designed to examine the effect of process parameters on the emulsification efficiency and stability index of Pickering emulsions. The emulsification efficiency is dominated by the JF concentration and length, while the emulsion stability could be prolonged through adjusting the JF concentration and hydrophilic graft density. JF-stabilized emulsions exhibit a much higher stability index (96.4%) than that of Janus microparticle counterparts (37.7%). Though there is no apparent effect on the surface wettability, JFs with PNIPAm grafts of about 2200 Da achieve the most stable Pickering emulsions. Superparamagnetic Fe3O4 nanoparticles are inoculated into JFs to collect emulsion droplets under a magnetic field, and the emulsions could be demulsified at an elevated temperature to harvest oil. Meanwhile, the recovered JF emulsifiers could be repeatedly used without loss of the emulsification efficiency. Thus, this study demonstrates surface-switchable JFs to be effective stabilizers of Pickering emulsions and readily recycled for oil harvesting from wastewater.

Photothermal Propelling and Pyroelectric Potential-Promoted Cell Internalization of Janus Nanoparticles and Pyroelectrodynamic Tumor Therapy.

Cancer phototherapy experiences limitations in tissue diffusion and cell internalization of phototherapeutic agents and dose-dependent side effects. Herein, Janus pyroelectric nanoparticles (NPs) are designed to generate self-powered motion and built-in electric fields to overcome the delivery barriers. Polydopamine (PDA) layers are partially coated on tetragonal BaTiO3 (tBT) NPs to prepare Janus tBT@PDA, and Au NPs are deposited on the PDA caps to obtain Janus tBT@PDA-Au NPs. Near-infrared (NIR) illumination of tBT@PDA-Au builds in situ pyroelectric potentials on NPs, which selectively affect the membrane potential of tumor cells rather than normal cells to enhance tumor cell internalization and produce reactive oxygen species (ROS) for pyroelectric dynamic therapy (PEDT). The asymmetric photothermal effect of the Janus NPs creates thermophoretic force to propel NP motion, which enhances tumor diffusion and cellular uptake of NPs and boosts cytotoxicity and intracellular ROS levels. The inoculation of Au NPs increases the photothermal effect, exhibits larger motion velocities, produces higher pyroelectric potentials, and elevates cellular uptake rates, resulting in significant induction of tumor cell apoptosis, suppression of tumor growth, and extension of animal survival. Thus, the concise design of tBT@PDA-Au/NIR treatment has achieved thermophoretic motion-promoted tissue diffusion, built-in electric field-enhanced cell internalization, and photothermal/PEDT-synergized antitumor efficacy.

Light-triggered theranostic hydrogels for real-time imaging and on-demand photodynamic therapy of skin abscesses.

The management of wound infection remains the major challenges in real-time diagnosis, effective bacterial elimination and rapid wound healing. Herein, we developed injectable theranostic hydrogels to achieve long-term visual imaging of infected wounds and possible infection recurrence and to launch an on-demand bactericidal effect without using any antibiotics. Antimicrobial peptide ε-polylysine (ePL)-derived hydrogels were prepared through the copolymerization of methacrylated ePL (mPL) and the conjugates with tetrakis(4-carboxyphenyl) porphyrin (mPL-TCPP) and phenol red (mPL-Pr). Light illumination of mPL-TCPP produces reactive oxidative species (ROS) to initiate free radical crosslinking into PL@Pr-TCPP hydrogels without using any additional photoinitiators and concurrently exhibits antibacterial photodynamic therapy (PDT). PL@Pr-TCPP hydrogels experience quick color changes from yellow to orange and finally to red when pH values change from 5.0 to 9.0. The actual pH and related bacterial levels in the wounds could be read from G/B signal ratios of hydrogel colors captured by a smart phone. The conjugation of phenol red and TCPP into hydrogels affords a robust bacterial infection diagnosis and persistent bactericidal effect after cycled light illumination. The bacterial capture by ePL hydrogels strengthens PDT effect through alleviating the short lifetime and action distance of ROS. On a Staphylococcus aureus-infected abscess model, light illumination of the pregel solutions achieves in situ formation of hydrogel dressings. The synergistic bactericidal performance significantly relieves inflammatory status, accelerates collagen deposition, and promotes neovascularization, leading to full recovery of the infected wounds with regeneration of skin accessories. PL@Pr-TCPP hydrogels on the wound bed show color changes upon the recurrence of bacterial infection, which could also be totally eliminated after light illumination. Therefore, this study demonstrates a feasible strategy to develop theranostic hydrogel dressings for life-cycle diagnosis and on-demand treatment of wound infections. STATEMENT OF SIGNIFICANCE: Over 30% of skin and soft tissue infections become chronic even after appropriate antibacterial treatment, and recurrent infections are commonly reported after initial infection. Challenges remain in the development of theranostic wound dressings having the capability of point-of-care diagnosis, life-cycle monitoring and on-demand elimination of bacterial infection. Herein, light-triggered gelation is used to develop theranostic hydrogels for reversible naked-eye diagnosis and on-demand photodynamic therapy of wound infections. Light illumination plays a "one-stone-two-birds" role, i.e., photodynamically produced reactive oxidative species enable bactericidal effect without using any antibiotics, and the generated free radicals initiate crosslinking of hydrogels without using any additional photoinitiators. Bacterial infection-activated color changes of hydrogels could be captured with a smart phone for on-site and persistent monitoring of bacterial infection and wound healing process.

Bacteria-Templated and Dual Enzyme-Powered Microcapsule Motors To Promote Thrombus Penetration and Thrombolytic Efficacy.

Antithrombotic therapy is confronted with short half-lives of thrombolytic agents and high bleeding risks. Challenges remain in the development of drug delivery systems for thorough destruction of thrombi and timely restoration of blood flow while minimizing side effects. Herein, polydopamine capsule-like micromotors with urokinase (uPA) loadings and Arg-Gly-Asp (RGD) grafts (r-u@PCM) were constructed using rod-shaped bacteria as the template, and one single opening was created on each capsule through bacterial ghost (BG) formation. Glucose oxidase and catalase were encapsulated in the large cavity of microcapsules, and their successive oxidation of glucose produced O2 bubbles, which ejected out through the single opening to propel the motion of r-u@PCM. In vitro targeting testing of r-u@PCM shows significant higher accumulations on the activated platelets than those without RGD grafts (u@PCM, 7 folds) or without enzyme loadings (r-u@PC, 11 folds). Compared with the major distribution of r-u@PC on the clot surface, r-u@PCM efficiently penetrates into clots with dense fibrin networks, and near-infrared (NIR) irradiation (r-u@PCM/NIR) promotes thrombus infiltration through increasing uPA release and thermolysis of the networks. Pharmacokinetic study shows that the loading of uPA in r-u@PCM extends the terminal half-life from 24 min to 5.5 h and the bioavailability increased 13 times. In a hindlimb venous thrombosis model, r-u@PCM/NIR treatment promotes uPA accumulations in thrombi and disrupts all the thrombi after 8 h with a full recovery of blood flows. Effective thrombolysis is also achieved even after reducing the uPA dose 5 times. Thus, this is the first attempt to fabricate rod-shaped microcapsule motors through a biologically derived method, including bacterial templating and BG formation-induced opening generation. r-u@PCM/NIR treatment promotes thrombolysis through the photothermal effect, self-propelled infiltration into thrombi, and accelerated local release of uPA, providing a prerequisite for reducing uPA dose and bleeding side effects.

Nitric oxide-propelled nanomotors for bacterial biofilm elimination and endotoxin removal to treat infected burn wounds.

Biofilm infection is regarded as a major contributing factor to the failure of burn treatment and a persistent inflammatory state delays healing and leads to the formation of chronic wounds. Herein, self-propelled nanomotors (NMs) are proposed to enhance biofilm infiltration, bacterial destruction, and endotoxin clearance to accelerate the healing of infected burn wounds. Janus nanoparticles (NPs) were prepared through partially coating Fe3O4 NPs with polydopamine (PDA) layers, and then polymyxin B (PMB) and thiolated nitric oxide (SNO) donors were separately grafted onto the Janus NPs to obtain IO@PMB-SNO NMs. In response to elevated glutathione (GSH) levels in biofilms, NO generation from one side of the Janus NPs leads to self-propelled motion and deep infiltration into biofilms. The local release of NO could destroy bacteria inside the biofilm, which provides a non-antibiotic antibiofilm approach without the development of drug resistance. In addition to intrinsic antibacterial effects, the PMB grafts preferentially bind with bacteria and the active motion enhances lipopolysaccharide (LPS) clearance and then significantly attenuates the production of inflammatory cytokines and reactive oxide species by macrophages. Partial-thickness burn wounds were established on mice and infected with P. aeruginosa, and NM treatment almost fully destroyed the bacteria in the wounds. IO@PMB-SNO NMs absorb LPS and remove it from the wounds under a magnetic field, which downregulates the interleukin-6 and tumor necrosis factor-α levels in tissues. The infected wounds were completely healed with the deposition and arrangement of collagen fibers and the generation of skin features similar to those of normal skin. Thus, IO@PMB-SNO NMs achieved multiple-mode effects, including GSH-triggered NO release and self-propelled motion, the NO-induced non-antibiotic elimination of biofilms and bacteria, and PMB-induced endotoxin removal. This study offers a feasible strategy, with integrated antibiofilm and anti-inflammatory effects, for accelerating the healing of infected burn wounds.

Persistent Luminescence-Based Theranostics for Real-Time Monitoring and Simultaneously Launching Photodynamic Therapy of Bacterial Infections.

External light irradiation is usually required in bacterial infection theranostics; however, it is always accompanied by limited light penetration, imaging interference, and incomplete bacterial destruction. Herein, a feasible "image-launching therapy" strategy is developed to integrate real-time optical imaging and simultaneous photodynamic therapy (PDT) of bacterial infections into persistent luminescence (PL) nanoparticles (NPs). Mesoporous silica NPs are used as a substrate for in situ deposition of PL nanodots of ZnGa2 O4 :Cr3+ to obtain mPL NPs, followed by surface grafting with silicon phthalocyanine (Si-Pc) and electrostatic assembly of cyanine 7 (Cy7) to fabricate mPL@Pc-Cy NPs. The PL emission of light-activated mPL@Pc-Cy NPs is quenched by Cy7 assembly at physiological conditions through the fluorescence resonance energy transfer effect, but is rapidly restored after disassembly of Cy7 in response to bacterial infections. The self-illuminating capabilities of NPs avoid tissue autofluorescence under external light irradiation and achieve real-time colorimetric imaging of bacterial infections. In addition, the afterglow of mPL NPs can persistently excite Si-Pc photosensitizers to promote PDT efficacy for bacterial elimination and accelerate wound full recovery with normal histologic features. Thus, this study expands the theranostic strategy for precise imaging and simultaneous non-antibiotic treatment of bacterial infections without causing side effects to normal tissues.

Bacteria-propelled microtubular motors for efficient penetration and targeting delivery of thrombolytic agents.

Effective thrombolysis is critical to rapidly rebuild blood flow for thrombosis patients. Drug delivery systems have been developed to address inadequate pharmacokinetics of thrombolytic agents, but challenges still remain in the timely removal of blood clots regarding the dense fibrin networks. Herein, rod-shaped tubular micromotors were developed to achieve efficient penetration and thorough destruction of thrombi. By using electrospun fiber fragments as the template, urokinase (uPA)-loaded polydopamine (PDA) microtubes with surface decorated fucoidan (FuPDAuPA) were prepared at the aspect ratio of around 2. One E. coli Nissle 1917 (EcN) was assembled into one microtube to construct a FuPDAuPA@EcN hybrid micromotor through PDA adhesion and L-aspartate induction. The pharmacokinetic analysis indicates that the encapsulation of uPA into micromotors extends the half-life from 0.4 to 5.6 h and increases the bioavailability over 10 times. EcN-propelled motion elevates adsorption capacities of FuPDAuPA@EcN for more than four times compared with that of FuPDAuPA. The fucoidan-mediated targeting causes 2-fold higher thrombolysis capacity in vitro and over 10-fold higher uPA accumulation in thrombi in vivo. In the treatment of venous thrombi at mouse hindlimbs, intravenous administration of FuPDAuPA@EcN completely removed blood clots with almost full recovery of blood flows and apparently alleviated tail bleeding. It should be noted that FuPDAuPA@EcN treatment at a reduced uPA dose caused no significant difference in the blood flow rate compared with those of FuPDAuPA. The synergistic action of fucoidan-induced targeting and EcN-driven motion provides a prerequisite for promoting thrombolytic efficacy and reducing uPA dose and bleeding side effect. STATEMENT OF SIGNIFICANCE: The standard treatment to thrombosis patient is intravenous infusion of thrombolytic agents, but the associated bleeding complications and impairment of normal haemostasis greatly offset the therapeutic benefits. Drug delivery systems have been developed to address the limitations of inadequate pharmacokinetics of thrombolytic agents, but challenges still exist in less efficient penetration into dense networks for thorough destruction of thrombi. Up to now only few attempts have been made to construct nano-/micromotors for combating thrombosis and there is no single case that antithrombosis is assisted by bacteria or cells-propelled motors. Herein, bacteria-propelled microtubes were developed to carry urokinase for efficient penetration into blood clots and effective thrombolysis. The synergistic action of bacteria-driven motion and specific ligand-induced targeting holds a promising treatment strategy for life-threatening cardiovascular diseases such as thrombosis and atherosclerosis.

Bacterial navigation for tumor targeting and photothermally-triggered bacterial ghost transformation for spatiotemporal drug release.

Cancer chemotherapy is confronted with challenges regarding the effective delivery of chemotherapeutics into tumor cells after systemic administration. Herein, we propose a strategy to load drugs into probiotic E. coli Nissle 1917 (EcN) for self-guided navigation to tumor tissues and subsequently release the drugs with in situ transformation into bacterial ghosts (BGs). Chemotherapeutic agent 5-fluorouracil (FU) and macrophage phenotype regulator zoledronic acid (ZOL) are loaded into EcN through electroporation, followed by decoration of Au nanorods on the ECN surface to construct EcNZ/F@Au. High loading levels of 5FU (8.8%) and ZOL (10.5%) are achieved as well as high retention rates of bacterial viability (87%) and motion velocity (88%). Under near infrared (NIR) illumination the photothermal effect of Au nanorods elevates the local temperature to induce the transformation of live EcN into BGs. The created transmembrane channels initiate the gradual drug release from BGs, thus representing the first attempt to control the drug release via a biological evolution. An intermittent NIR illumination causes stepwise increases in the BG formation and drug release, which could implement an external on-off control and spatiotemporal drug release. Self-guided motion of EcN promotes efficient extravasation across blood vessels and preferential accumulation of drugs in tumors. In addition to the chemotherapeutic effect of FU, the local release of ZOL from EcNZ/F@Au enhances valid polarization of tumor-associated macrophages toward the M1 phenotype and an effective production of proinflammatory cytokines, leading to a synergistic efficacy on tumor growth inhibition. Thus, this study demonstrates a feasible strategy to integrate chemotherapy, immunotherapy, and photothermal effects in a concise manner for effective cancer treatment with few side effects. STATEMENT OF SIGNIFICANCE: Bacteria are capable to trace and colonize in hypoxic tumor tissues. Bacterial drug carriers indicate limitations in efficient drug loading and effective release modulation. Herein, we propose a strategy to load drugs into bacteria for self-guided delivery and subsequently release the drugs in tumors with in situ transformation into bacterial ghost (BGs). Drugs are loaded into live bacteria through electroporation and Au nanorods are decorated on the bacterial surface, wherein the photothermal effect, chemotherapy, and immunotherapy are integrated in a concise manner. NIR illmumination of Au nanorods elevates the local temparature, induces the BG tranformation, and activates the spatiotemporal drug release, representing the first attempt of release modulation via a biological evolution.

Cell Adhesion-Mediated Piezoelectric Self-Stimulation on Polydopamine-Modified Poly(vinylidene fluoride) Membranes.

Cell adhesion-mediated piezoelectric stimulation provides a noninvasive method for in situ electrical regulation of cell behavior, offering new opportunities for the design of smart materials for tissue engineering and bioelectronic medicines. In particular, the surface potential is mainly dominated by the inherent piezoelectricity of the biomaterial and the dynamic adhesion state of cells. The development of an efficient and optimized material interface would have important implications in cell regulation. Herein, we modified the surface of poled poly(vinylidene fluoride) (PVDF) membranes through polymerization of dopamine and investigated their influence on cell adhesion and electromechanical self-stimulation. Our results demonstrated that mesenchymal stem cells seeded on the poled PVDF membrane exhibited stronger cell spreading and adhesion. Meanwhile, the surface modification through polydopamine significantly improved the hydrophilicity of the samples and contributed to the formation of cell actin bundles and maturation of focal adhesions, which further positively modulated cell piezoelectric self-stimulation and induced intracellular calcium transients. Combining with theoretical simulations, we found that the self-stimulation was enhanced mainly due to the increase of the adhesion site and adhesion force magnitude. These findings provide new insights for probing the cell regulation mechanism on piezoelectric substrates, offering more opportunities for the rational design of piezoelectric biomaterial interfaces for biomedical engineering.

Icebreaker-inspired Janus nanomotors to combat barriers in the delivery of chemotherapeutic agents.

Cancer chemotherapy remains challenging to pass through various biological and pathological barriers such as blood circulation, tumor infiltration and cellular uptake before the intracellular release of antineoplastic agents. Herein, icebreaker-inspired Janus nanomotors (JMs) are developed to address these transportation barriers. Janus nanorods (JRs) are constructed via seed-defined growth of mesoporous silica nanoparticles on doxorubicin (DOX)-loaded hydroxyapatite (HAp) nanorods. One side of JRs is grafted with urease as the motion power via catalysis of physiologically existed urea, and hyaluronidase (HAase) is on the other side to digest the viscous extracellular matrices (ECM) of tumor tissues. The rod-like feature of JMs prolongs the blood circulation, and the self-propelling force and instantaneous digestion of hyaluronic acid along the moving paths promote extravasation across blood vessels and penetration in tumor mass, leading to 2-fold higher drug levels in tumors after JM administration than those with JRs. The digestion of ECM in the diffusion paths is more effective to enhance drug retention and diffusion in tumors compared with enzyme-mediated motion. The ECM digestion and motion capabilities of JMs show no influence on the endocytosis mechanism, but lead to over 3-fold higher cellular uptake than those of pristine JRs. The JM treatment promotes therapeutic efficacy in terms of survival prolongation, tumor growth inhibition and cell apoptosis induction and causes no tumor metastasis to lungs with normal alveolar spaces. Thus, the self-driven motion and instantaneous clearance of diffusion routes demonstrate a feasible strategy to combat a series of biological barriers in the delivery of chemotherapeutic agents in favor of antitumor efficacy.

Shape switching of CaCO3-templated nanorods into stiffness-adjustable nanocapsules to promote efficient drug delivery.

Geometry and mechanical property have emerged as important parameters in designing nanocarriers, in addition to their size, surface charge, and hydrophilicity. However, inconsistent and even contradictory demands regarding the shape and stiffness of nanoparticles have been noted in blood circulation, tumor accumulation, and tumor cell internalization. Herein, CaCO3 nanorods (NRs) with an aspect ratio of around 2.4 are assembled with hyaluronic acid (HA) hydrogel layers to prepare CaCO3@HA NRs. The rod geometry enables lower recognition by macrophages and higher extravasation into tumor tissues than the spherical counterpart. In response to the slightly acidic tumor matrix, the acid-labile removal of CaCO3 templates achieves shape switching into spherical HA nanocapsules (NCs). The shape switchable CaCO3@HA NRs show significantly higher uptake and cytotoxicities to 4T1 cells than CaCO3-Si@HA NRs with silica layers on CaCO3 cores to inhibit shape switching. In addition, HA NCs with 2 - 8 layers of HA hydrogels exhibit stiffness from 1.85 to 12.3 N/m, and the assembly of 4 layers shows 2- to 3-fold higher cellular uptake than those of other NCs. The shape shift satisfies long-term blood circulation of NRs, and the resulting stiffness-adjustable NCs promote tissue infiltration and intracellular accommodation, resulting in a 4-fold higher drug accumulation in tumors. The CaCO3@HA NR treatment significantly suppresses tumor growth; prolongs animal survival; inhibits lung metastasis; and eliminates systemic toxicities to blood, liver, kidney, and heart tissues. This study achieves a comprehensive understanding of the shape and stiffness effects and demonstrates a hierarchical targeting strategy to address the multiple delivery barriers for chemotherapeutic agents. STATEMENT OF SIGNIFICANCE: The different barriers involved in the drug delivery pathway have inconsistent and even contradictory demands on the shape and stiffness of nanoparticles. In the current study, in situ switching of nanorods (NRs) into spherical nanocapsules (NCs) in tumor tissues is proposed to address these dilemmas. The NR shape ensures long-term blood circulation and high tumor tissue accumulation, while the in situ switching into NCs promotes tissue infiltration and cellular internalization. NCs with different numbers of hydrogel layers also provide a robust system wherein NC stiffness is controlled as a single variable to study stiffness-dependent cellular behaviors. Thus, this straightforward design offers a comprehensive understanding of how the shape and stiffness of nanocarriers affect their biological pathways.

The Effect of Xinmailong Infusion on Sepsis-Induced Myocardial Dysfunction: a Pragmatic Randomized Controlled Trial.

ABSTRACT: Sepsis-induced myocardial dysfunction (SIMD) contributes significantly to cardiovascular dysfunction during septic shock. We aimed to evaluate the potential role of Xinmailong injection (XMLI), a polypeptide medicine extracted from Periplaneta americana, in reversing the progression of myocardial damage to SIMD in sepsis patients. This was a multicenter, randomized, double-blind, parallel-group trial. We recruited all patients consecutively admitted to intensive care units (ICUs) who were aged 18 to 85 years old and met the sepsis 3.0 criteria. The primary outcome measure was the incidence of sepsis-induced myocardial dysfunction while in the ICU. Of the 192 patients, 96 were assigned to the treatment group, and 96 to the control group. Subsequently, 41 patients [41/96 (42.7%)] in the XMLI group and 61 patients in the placebo group [61/96 (63.5%)] were confirmed to have diastolic dysfunction on the fifth day (D5). The incidence of diastolic SIMD was significantly different between the two groups (P = 0.004). There were 36 deaths in the two groups during the 28-day follow-up, with a general mortality rate of 18.8% (36/192). The 28-day mortality rates were not significantly different between the groups (P = 0.45). However, the brain natriuretic peptide (BNP) plasma concentration trends on D0, D2, and D5 significantly differed between the two groups (P = 0.049). In septic patients, XMLI decreased the occurrence rate of diastolic SIMD more effectively than the placebo. The improvement in serum BNP concentration was also greater in the XMLI group. XMLI may, therefore, effectively and safely improve cardiac function in patients with sepsis.

Hierarchically targetable fiber rods decorated with dual targeting ligands and detachable zwitterionic coronas.

The shapes of drug carriers have significant effects on the drug's blood circulation lifetime and tumor accumulation levels. In this study, nonspherical drug carriers of fiber rods are enhanced with hierarchically targeting capabilities to achieve long circulation in blood, on-demand recovery of cell targeting ligands in tumor tissues and dual ligands-mediated cellular uptake. Zwitterionic polymers are conjugated on fiber rods via acid-labile linkers as stealth coronas to reduce the capture by macrophages and shield the targeting ligands. Compared with commonly used poly(ethylene glycol), the zwitterionic grafts show significantly higher inhibition of protein adsorption and lower internalization by macrophages, leading to around 2 folds longer blood circulation and over 2.5 folds higher drug accumulation in tumors than pristine fiber rods. To address the conflicts between blood circulation and cellular uptake, the zwitterionic coronas are efficiently removed in the slightly acidic tumor microenvironment. The exposure of targeting ligands could activate the internalization by tumor cells, resulting in higher cytotoxicity and tumor accumulation than those with stable linkers. Fiber rods are grafted with dual ligands of folate and biotin, and the optimal ligand densities and ratios are determined to maximize the tumor cell uptake. Compared with other treatment, fiber rods with decorated zwitterionic coronas and acid-liable exposure of dual targeting ligands enhance the suppression of tumor growth, prolong animal survival, and cause less lung metastasis. The development of fiber rods with hierarchically targeting capabilities shows great potential in improving the blood circulation, tumor accumulation and cellular uptake, and eventually promoting therapeutic efficacy. STATEMENT OF SIGNIFICANCE: The targeted delivery of chemotherapeutic agents will encounter a series of biological and pathological barriers. In this study, fiber rods were empowered with hierarchically targeting capabilities to resolve the conflict between blood circulation and cellular uptake. This strategy has shown several advantages over the existing methods. Firstly, zwitterionic polymers were used as blood circulation ligands, and concrete evidence was provided via head-to-head comparison with commonly used poly(ethylene glycol) ligands in the macrophage uptake and in vivo tissue distribution. Secondly, the depletion of circulation ligands and on-demand exposure of targeting ligands in tumor tissues showed crucial effects on the uptake by tumor cells. Thirdly, the densities and ratios of the dual targeting ligands were initially determined for a maximal cellular internalization.

Intracellular bacteria destruction via traceable enzymes-responsive release and deferoxamine-mediated ingestion of antibiotics.

Intracellular bacteria (ICBs) are among the most life-threatening causes of drug resistance. Challenges remains in the intracellular drug release specific to ICB-infected cells and efficient uptake into ICBs. In this study, mannose-grafted polymers containing enzymes-responsive and tetraphenylethylene segments (mPET) are assembled into nanoparticles with loading complexes of deferoxamine-ciprofloxacin conjugates with Fe3+ (DFeC). The aggregation-induced emission (AIE) of tetraphenylethylene segments is overlapped with DFeC absorptions, leading to fluorescence resonance energy transfer (FRET)-caused quenching of mPET@DFeC nanoparticles. Nanoparticles are efficiently acquired by infected macrophages via mannose mediation, and the DFeC release is triggered by intracellular lipase and alkaline phosphatase specific to ICB-engulfed macrophages, followed by deferoxamine-mediated ingestion of ciprofloxacin into ICBs. The gradual alleviation of FRET effect and the concurrent restoration of AIE activity demonstrate capabilities of dynamically tracking the drug release and ICB treatment outcome. The mPET@DFeC treatment inhibits the hematological, hepatic and nephric toxicities caused by ICB infections, and all the infected mice survive with dramatic reductions of bacterial levels in livers (over 430 folds), spleens (over 240 folds) and kidneys (5.6 × 104 folds). Thus, this study has provided a feasible strategy to achieve intracellular enzymes-responsive and traceable release of antibiotics and then deferoxamine-mediated bacterial ingestion for ICB destruction.

Fibrous testing papers for fluorescence trace sensing and photodynamic destruction of antibiotic-resistant bacteria.

The increasing prevalence of antibiotic-resistant bacteria needs rapid identification and efficient destruction routes. This study proposes testing paper derived from electrospun fibrous mats and aggregation-induced emission (AIE) probes for trace sensing and simultaneous destruction of antibiotic-resistant E. coli. Aptamers are conjugated on fibers for selective capture of E. coli, and the capture capability can be regenerated via rinsing with salt solution. Hydroxyl tetraphenylethene (TPE) is linked with two cephalosporin molecules to construct TPE-Cep probes, and the fluorescence emission is turned on specifically in the presence of ß-lactamase, which is a critical marker for screening resistant bacteria. Fibrous mats are lit up only in the presence of antibiotic-resistant bacteria, and the fluorescence intensity changes could be statistically fitted into an equation for quantitative analysis. Fibrous strips display apparent color changes from blue to green for a visual readout of bacterial levels, and the limit of detection (LOD) is much lower than those of previous paper substrates. In addition, the TPE-Cep probes could produce reactive oxygen species (ROS) under room light illumination to kill the captured bacteria. Thus, the integration of aptamer-grafted electrospun fibers and functional AIE probes provides potential for selective capture, trace imaging and photodynamic destruction of antibiotic-resistant bacteria.

Fibrous strips decorated with cleavable aggregation-induced emission probes for visual detection of Hg2.

The widespread contamination and high poisonousness have created significant concerns and thus demands for facile, rapid and selective monitoring of trace Hg2+. Inspired from the unique aggregation-induced emission (AIE) feature, in the current study, novel tetraphenylethylene (TPE) derivatives are prepared containing sulfonic groups for water solubility modulation and carboxyl dithioacetals for Hg2+ sensing. The TPE derivatives are grafted on electrospun fiber as test papers to initiate the AIE activities, while the Hg2+-specific cleavage of dithioacetal groups leads to the release of TPE derivatives and fluorescence turn-off. The decrease in the fluorescence intensities of fibrous mats could be fitted with Hg2+ levels for quantitative analysis, and the fibrous mats turn from green to bluish-green and then to blue in the presence of different Hg2+ levels. The limit of detection (LOD) reaches as low as 20 nM Hg2+, satisfying the threshold detection in drinking water, and the Hg2+ sensing indicates negligible interference from other metal ions and pH variations. The detected Hg2+ levels in lake water are consistent with the added amount with a recovery rate of over 98 %. It demonstrates a feasible strategy to integrate Hg2+-cleavable AIE probes on fibrous strips for real-time, highly specific and naked-eye detection of trace Hg2+.

Comparison of revised EWGSOP criteria and four other diagnostic criteria of sarcopenia in Chinese community-dwelling elderly residents.

OBJECTIVES: To compare the prevalence and associated factors of sarcopenia defined by the revised European Working Group on Sarcopenia in Older People (EWGSOP2) criteria with the initial European Working Group on Sarcopenia in Older People (EWGSOP1) criteria, the Asia Working Group for Sarcopenia (AWGS), the International Working Group on Sarcopenia (IWGS), and the National Institutes of Health (FNIH) Sarcopenia Project criteria among Chinese community-dwelling older adults. DESIGN: A cross-sectional study. SETTING: Two community health centers in Urumqi, China. PARTICIPANTS: A total of 483 participants aged 60 years and older from the community. MEASUREMENT: Anthropometry, skeletal muscle mass, handgrip strength, 4-m walking speed, and biochemical markers. Questionnaire collected information included demographics, lifestyle, and quality of life. RESULTS: The prevalence of EWGSOP2-defined sarcopenia (men: 6.5%; women: 3.3%) was lower than that defined by the EWGSOP1 (men: 22.3%; women 11.7%), AWGS (men: 10.9%; women: 8.0%), and IWGS (men: 24.5%; women: 11.0%) criteria, but higher than FNIH criteria (men: 6.0%; women: 1.7%). The positive percent agreement was lower (men: 15.6%-63.6%; women: 15.2%-40.0%), while negative percent agreement was higher (men: 96.4%-100.0%; women: 97.3%-99.6%). Sex (OR 0.31, 95% CI 0.12-0.81), education level (OR 0.49, 95% CI 0.29-0.83), and body mass index (BMI, OR 0.73, 95% CI 0.62-0.86) were associated with sarcopenia defined by the EWGSOP2 criteria. No consistent pattern of risk factors associated with sarcopenia in EWGSOP2 and four other diagnostic criteria was present. CONCLUSIONS AND IMPLICATIONS: The EWGSOP2 criteria did not agree with the EWGSOP1, AWGS, IWGA, and FNIH criteria defining sarcopenia. Risk factors associated with the EWGSOP2-defined sarcopenia have no consistent patterns with the EWGSOP1, AWGS, IWGA, and FNIH criteria. Therefore, the validity of the EWGSOP2 consensus needs to be confirmed in further prospective studies.