A facile aptamer immobilization strategy to fabricate a robust affinity monolith for highly specific in-tube solid-phase microextraction.

Developing a functional affinity monolithic column towards in-tube solid-phase microextraction (IT-SPME) for selective sample pretreatment is critical. Herein, a high-performance capillary affinity monolithic column with an ultra-high aptamer coverage density was rapidly fabricated via a simple adsorption strategy, in which aptamers with natural sequences were directly immobilized on an ammonium-based strongly cationic matrix. Limitations of the traditional biological or covalent methods such as time-consuming modification reactions, special requirement of active groups (e.g. -NH2 and -SH) on the aptamer, and low aptamer coverage density levels were avoided. An ultra-high coverage density of 8616 pmol µL-1 was achieved with excellent stability, and the highest aptamer-modification level among all the current methods was reached. Selective recognition and high recovery yields of the model mycotoxin ochratoxin A (OTA) were achieved in 95.9 ± 0.98%-97.9 ± 0.28% (n = 3). In particular, there was little cross-reactivity towards the OTB analogue of only 0.5% even in the case of 250 fold of the analogue OTB, which was not reported in previous affinity monoliths. Upon sample analysis, satisfactory discriminations of trace OTA were obtained at 93.7 ± 1.4%-95.5 ± 2.5% (n = 3) in beer and wheat. A facile and direct method for efficiently fabricating an aptamer-based affinity monolith towards online selective IT-SPME was proposed.

In situ photo-initiated polymerized oligonucleotide-functionalized hydrophilic capillary affinity monolith for highly selective in-tube microextraction of ochratoxin A mycotoxin.

A photo-initiated polymerized oligonucleotide-grafted hydrophilic affinity monolithic column was synthesized in situ, and exploited for selective in-tube solid phase micro-extraction (IT-SPME) protocol towards the sensitive detection of ochratoxin A (OTA). Only 7 min was required for the rapid polymerization of aptamer-based affinity monolith, which was much less than the reaction time of most thermal polymerization (12-16 h) and sol-gel chemistry methods (up to 52 h). Characterizations such as polymerization recipes, structure morphology, FTIR spectrum, elemental mapping, mechanical stability, and specific recognition performance were evaluated. A significantly hydrophilic nature with a low contact angle of 15° was observed, and a mixed-mode mechanism including aptamer affinity recognition and hydrophilic interaction (HI) was employed. By coupling with HPLC-fluorescence detection, the highly specific online recognition performance was achieved with an extremely low nonspecific adsorption of the analogues. The calibration curve of OTA was obtained in the concentration range 0.05-50.00 ng·mL-1 with a limit of detection (LOD, S/N = 3) of 0.012 ng·mL-1. Applied to sample analysis, acceptable recovery yields of 95.1 ± 1.4% - 99.5 ± 2.2% (n = 3) were obtained in beer and red wine. The proposed method lighted a promising way to efficiently preparing a hydrophilic aptamer-affinity monolith for highly specific recognition of trace mycotoxin by IT-SPME coupled with HPLC. A hydrophilic oligonucleotide-based affinity capillary monolith was explored via in situ photopolymerization for overcoming low preparation efficiency and achieving high-performance online IT-SPME of OTA mycotoxin.

Facile preparation of stainless steel microextraction fiber via in situ growth of metal-organic framework UiO-66 and its application to sensitive analysis of polycyclic musks.

A functional stainless steel microextraction fiber easily prepared by in situ growing metal-organic framework UiO-66 was presented and used for high-performance analysis of polycyclic musks. Via the robust Ag-SH bonding reaction, mercaptoacetic acid was easily anchored on Ag film to provide carboxyl group on the stainless steel fiber, then in situ grown UiO-66 was fulfilled via the coordination reaction between Zr4+ and carboxyl group. Good characteristics including large surface area, high thermal stability, and good adsorption property were achieved. Sensitive detection limits (0.015-0.040 ng/L) were achieved for polycyclic musks by coupling with gas chromatography with mass spectrometry, and it could be stable enough for 150 extraction cycles without a significant loss of extraction efficiency. Compared with the classical commercial fibers, 2.2-11.4 times higher enhancement factors were shown. Applied to the analysis of fortified river water samples, five typical polycyclic musks were well detected with the recoveries of 90.2-101.8%, respectively. It showed a facile approach for preparing stainless steel microextraction fiber via chemically bonding in situ grown metal-organic framework for high-performance enrichment.

Sensitive amperometric detection for capillary electrophoresis of phenol carbamates with in-line thermal hydrolysis strategy.

A strategy on amperometric detection for CZE of phenol carbamates as model analytes with a facile in-line thermal hydrolysis was presented, in which a thermal hydrolysis, subsequent CZE separation and final column-end amperometric detection were accomplished in an intact capillary. Key parameters of hydrolysis dynamics of carbamates and electrochemical detection of the hydrolysates were studied, as well as electrophoretic conditions. Under the optimal conditions, the capillary was utilized as chambers for in situ hydrolysis, CZE separation, and electrochemical detection. The successive separation of hydrolysates of five carbamates (propoxur, carbofuran, 3-OH-carbofuran, carbaryl and bendiocarb) were achieved within 17 min. Applied to vegetable samples, the recoveries of carbamates fortified at 0.02 and 0.05 mg/kg were ranging in 88-107.2 and 86.3-107.3%, respectively. The success in the implementation of such a scheme resulted in a simple instrument as compared with those current analytical methods with post-column derivization or pre-column hydrolysis, or online enrichment in chip, respectively. This protocol might possess a potential utility for the sensitive amperometric detection of phenol carbamates.

Highly hydrophilic polyhedral oligomeric silsesquioxane (POSS)-containing aptamer-modified affinity hybrid monolith for efficient on-column discrimination with low nonspecific adsorption.

A novel polyhedral oligomeric silsesquioxane (POSS)-containing aptamer-modified hybrid affinity monolith with excellent hydrophilicity and unique architecture without Si-OH groups is presented herein, and the nonspecific adsorption caused by the hydrophobic nature of the monolithic column or polar interaction with silanol groups is minimized. Via a simple "one-pot" procedure, hydrophilic monomers were facilely polymerized with the POSS-methacryl substituted (POSS-MA) and aptamer; a highly hydrophilic nature was obtained and the lowest contact angle of 11° was achieved. By using ochratoxin A (OTA) as the model analyte, highly selective recognition of OTA in the mixture was achieved and the control of nonspecific interactions and the cross-reactivity of OTB and AFB1 were significantly improved. The recovery yield of OTB caused by nonspecific adsorption in the resultant monolith was only about 0.1% and remained steady even with the coexistence of a high OTB content (OTA : OTB = 1 : 50), which reached the best level to date and was obviously less than the 6.1% occurring in the hydrophobic POSS-containing control monolith, 8.3% in the POSS-PEI@AuNPs@aptamer affinity monolith and 18.7% in the silica-hybrid affinity monolith. When applied to wine and wheat samples, the nonspecific adsorption was significantly reduced and efficient discrimination of OTA was obtained with better results than that of the hydrophobic POSS-containing affinity column. This provides an attractive tool for minimizing the nonspecific adsorption for highly selective on-column recognition.

Preparation of aptamer-bound polyamine affinity monolithic column via a facile triazine-bridged strategy and application to on-column specific discrimination of ochratoxin A.

Developing a high-performance modification protocol is critical for efficiently fabricating affinity monolith. Herein, by using 2,4,6-trichloro-1,3,5-triazine as the linker, a simple triazine-bridged approach was proposed for efficiently fabricating aptamer-grafted polyhedral oligomeric silsesquioxane-polyethyleneimine affinity monolith with high specificity toward ochratoxin A. Experimental parameters, column characteristics and specificity performances of the resultant affinity monolith were investigated in detail. Under the optimal conditions, 2,4,6-trichloro-1,3,5-triazine was rapidly grafted on the polyamine matrix, and effectively applied to the subsequent bridge linkage of aptamers. It was simple and effective, which resulted in a significant decrease of modification time, excellent properties including the high coverage density of aptamer up to 1799 pmol/µL and sensitive detection of ochratoxin A as low as 10 pg/mL in beer samples. This protocol provides a facile approach for fabricating aptamer-grafted polyamine affinity monoliths with highly selective discrimination performance.

Quinine-modified polymer monolithic column with reversed-phase /strong anion-exchange mixed-mode for pressurized capillary electrochromatography.

Via the facile ring-opening reaction of epoxy groups with quinine, a novel polymer monolith with quaternary ammonium for reversed-phase/strong anion-exchange mixed-mode has been fabricated for pressurized capillary electrochromatography (pCEC). Optimization on the preparation of quinine-modified monoliths has been investigated, and characteristics including morphology, permeability, mechanical stability, reproducibility, and column performance have been also studied. Active quaternary ammonium groups were conveniently produced to generate cationic action sites and stable anodic electroosmotic flow. Multiple interactions including reversed-phase, strong anion-exchange, electrostatic repulsion and π-π stacking interactions were obtained. Satisfactory separation capability of various analytes such as alkylbenzenes, polycyclic aromatic hydrocarbons, benzoic acid and its homologs, and ß2 -receptor excitants has been achieved. Applied to the real sample, the good resolution of three alkaloids in Corydalis yanhusuo were achieved by pCEC with the quinine-modified monolith. The results light a potential access to facilely fabricating quaternary ammonium-functionalized polymer monolith with multiple interactions for efficient electrochromatography profiling of various compounds.

A facile AuNPs@aptamer-modified mercaptosiloxane-based hybrid affinity monolith with an unusually high coverage density of aptamer for on-column selective extraction of ochratoxin A.

A convenient and high-performance AuNPs@aptamer-modified mercaptosiloxane-based hybrid affinity monolithic column with an unusually high coverage density of aptamers was facilely prepared and used for on-column selective recognition of ochratoxin A (OTA). Due to the high surface-to-volume ratio of AuNPs, the robust conjugation of Au-SH and large specific surface area of hybrid-silica monolith, high coverage density of 5'-SH-aptamers up to 3494 pmol µL-1 was achieved, which was 2.5-10 folds higher than that of other previously reported affinity monoliths modified with AuNPs@Apt. Using OTA as the model analyte, the highly selective recognition of OTA was carried out via online coupling with HPLC, and the cross-reactivity towards analogues, such as OTB and aflatoxin B1, was weak. High recovery yields of OTA were achieved at more than 92% (n = 3) even when OTB was added at a high concentration level up to 50 ng mL-1. For sample analysis, efficient discrimination of OTA was successfully obtained with a sensitive detection limit of 25 pg mL-1. The recoveries of OTA with different fortified levels were achieved at 88.6%-94.1% and 88.2%-94.3% for beer and wine samples, respectively. This protocol provides a facile approach for fabricating a desirable affinity monolith modified with abundant aptamers for highly selective and sensitive on-column extraction of target analyte OTA.

Multivariate surface self-assembly strategy to fabricate ionic covalent organic framework surface grafting monolithic sorbent for enrichment of aristolochic acids prior to high performance liquid chromatography analysis.

Herein, an ionic covalent organic framework (iCOF) surface grafting monolithic sorbent was prepared by the multivariate surface self-assembly strategy for in-tube solid-phase microextraction (SPME) of trace aristolochic acids (AAs) in serum, traditional Chinese medicines (TCMs) and Chinese patent drug. Via adjusting the proportion of ionic COF building block during the self-assembly, the density of quaternary ammonium ions in the iCOF was modulated for the enhanced adsorption of AAs. The successful preparation of iCOF surface grafting monolithic sorbent was confirmed by different means. A multiple mode mechanism involving π-π stacking, hydrophobic, electrostatic and hydrogen-bonding interactions was primarily attributed to the adsorption. Several in-tube SPME operating conditions, such as the dosage of ionic COF building block, ACN percentage and TFA percentage in the sampling solution, ACN percentage and TFA percentage in eluent and the collection time span, were optimized to develop the online in-tube SPME-HPLC method for analysis of AAs. Under the optimized conditions, a good linearity was obtained in the concentration range of 20-1000 ng/mL for target AAs in serum samples, the limits of detection (LODs) were less than 10 ng/mL, while the recoveries ranged from 90.3 % to 98.7 % with RSDs (n = 5) below 7.9 %. This study developed a feasible approach to iCOF functionalized monolithic sorbent for SPME and further exhibited the vast potential for the application of COF based monolithic sorbent in sample preparation.

Versatile, reusable and highly sensitive SERS-based point-of-care testing microplatform for reliable ATP detection.

The advancement in miniaturized Raman spectrometers, coupled with the single-molecule-level sensitivity and unique fingerprint identification capability of surface-enhanced Raman scattering (SERS), offers great potential for point-of-care testing (POCT). Despite this, accurately quantifying analyte molecules, particularly in complex samples with limited sample volumes, remains difficult. Herein, we present a versatile and reusable SERS microplatform for highly sensitive and reliable quantitative detection of adenosine triphosphate (ATP) in biological fluids. The platform utilizes gold-Prussian blue core-shell nanoparticles modified with polyethyleneimine (Au@PB@PEI NPs), embedded within gold nanoparticle-immobilized capillary-based silica monolithic materials. PB acts as an internal standard, while PEI enhances molecular capture. The periodic, bimodal porous structure of the silica monolithic materials provides uniform and abundant sites for nanoparticle attachment, facilitating rapid liquid permeation, intense SERS enhancement, and efficient enrichment. The platform regulates ATP capture and release through magnesium ions in the liquid phase, eliminating matrix interferences and enabling platform reuse. Integrating efficient molecular enrichment, separation, an interference-free internal standard, a liquid flow channel, and a detection chamber, our platform offers simplicity in operation, exceptional sensitivity and accuracy, and rapid analysis (∼10 min). Employing PB as an internal calibration standard, ratiometric Raman signals (I732/I2123) facilitate precise ATP quantification, achieving a remarkable limit of detection down to 0.62 pM. Furthermore, this platform has been proven to be highly reproducible and validated for ATP quantification in both mouse cerebrospinal fluid and human serum, underscoring its immense potential for POCT applications.

A DNA machine-based magnetic resonance imaging nanoprobe for in vivo microRNA detection.

In situ monitoring microRNA (miRNA) expression in vivo holds immense potential for directly visualizing the occurrence and progression of tumors. However, the significant barrier to developing a probe that can overcome the low abundance of miRNAs while providing an output signal with unlimited tissue penetration depth remains formidable. In this study, we developed a DNA machine-based magnetic resonance imaging nanoprobe (MRINP) for amplified detection of miR-21 in vivo. The MRINP was constructed with superparamagnetic Fe3O4 nanoparticles (NPs), paramagnetic Gd-DOTA complexes, and miR-21-activated DNA machines; the DNA machine was composed of hairpin DNAzyme (HD) strands serving as the DNAzyme walker and hairpin substrate (HS) strands serving as the track. Once uptake into tumor cells, the intracellular miR-21 specifically recognized and hybridized with the HD strand, restoring the activity of DNAzyme. Subsequently, the DNAzyme walker autonomously traveled on the surface of MRINP, and each step movement of the DNAzyme walker resulted in the cleavage of its substrate strands and the ensued release of the Gd-DOTA complex-labeled oligonucleotides, turning on the T1 signal of Gd-DOTA complexes for in situ imaging of miR-21 in tumor-bearing mice. This strategy would offer a promising approach for mapping tumor-specific biomarkers in vivo with unlimited penetration depth.

Dipyridyl-immobilized ionic liquid type hybrid silica monolith for hydrophilic interaction electrochromatography.

A pyridinium-based immobilized ionic liquid type multifunctional hybrid silica monolith was prepared by the in situ polymerization of 3-chloropropyl-silica matrix and 4,4'-dipyridyl for hydrophilic interaction CEC. The obtained hybrid monolith possessed of high stable skeletal microstructures with obviously hydrophilic retention mechanism under ACN content >50% in the mobile phase. Strong and stable anodic EOF could be observed under a broad pH range from pH 3.0 to 9.0. Due to the immobilized dipyridyl groups bonded to the silica matrix surface, the resulting hydrophilic hybrid monolith possessed multiple separation interactions including hydrogen bond, π-π, and anion exchange. Excellent separations of various polar analytes including electroneutral phenols, charged acid nucleotides, and basic analytes were successfully achieved. The highest column efficiencies up to 120,000, 164,000, and 106,000 plates/m were obtained for nucleotides, nucleic acid bases, and nucleosides and nicotines, respectively. These results demonstrated that the dipyridyl-immobilized ionic liquid functionalized hybrid monolith possessed highly mechanical stability and good chromatographic performance for hydrophilic interaction electrochromatography.

Phenylalanine functionalized zwitterionic monolith for hydrophobic interaction electrochromatography.

A novel phenylalanine (Phe) functionalized zwitterionic monolith for hydrophobic electrochromatography was prepared by a two-step procedure involving the synthesis of glycidyl methacrylate based polymer monolith and subsequent on-column chemical modification with Phe via ring-opening reaction of epoxides. Benefitting from the hydrophobicity of both methacrylate-based matrix and aromatic group of Phe, this monolith could exhibit good hydrophobic interaction for the separation. Typical RP chromatographic behavior was observed toward various solutes. The well-controlled cathodic or anodic EOF of the prepared column could be facilely switched by altering the pH values of running buffers. The separation mechanism of this Phe functionalized zwitterionic monolith is discussed in detail. Two mixed-mode mechanisms of RP/cation exchange and RP/anion exchange could be further realized on the same monolith in different pH condition of the mobile phase. Versatile separation capabilities of neutral, basic, and acidic analytes have been successfully achieved in this zwitterionic monolith by CEC method.

Sensitive capillary electrophoretic profiling of nicotine and nornicotine in mushrooms with amperometric detection.

A highly sensitive capillary electrophoretic profiling of nicotine (NIC) and nornicotine (NNIC) was developed and applied to mushrooms with amperometric detection (AD). Effects of the experimental factors including detection potential, separation parameters, and sample pretreatment conditions were investigated. Under the optimal conditions, the electrophoretic analysis of NIC and NNIC was achieved within 8 min on a pencil carbon disc working electrode at 0.95 V, which was lower than those reported previously. Good calibration curves were obtained in 0.01-2.0 µg/mL and 0.02-3.0 µg/mL with the LOD of 2 ng/mL and 5 ng/mL for NIC and NNIC, respectively. The feasibility of the resultant method was verified. Average recoveries of different fortified levels ranged in 80.7-86.0% and 94.0-98.6% for NNIC and NIC were gained, respectively. Applied to a range of mushrooms (Boletus edulis and Lentinus edodes), the NIC contained naturally was successfully found in the level of 19.71-79.20 µg/kg. The results obtained with CE-AD method were acceptable and close to that of HPLC-MS.

Polyhedral oligomeric silsesquioxane (POSS)-based multifunctional organic-silica hybrid monoliths.

A facile polyhedral oligomeric silsesquioxane (POSS)-based hybrid monolith with multiple mechanisms was developed by an in situ polymerization. High mechanical stability and good separation capabilities to polar and hydrophobic analytes were successfully achieved. An ideal versatile organic-silica hybrid monolith was presented for easy access to the efficient separation of various analytes.

Rapid capillary electrochromatographic profiling of phytohormones on a hydrophilic interaction/strong anion-exchange mixed-mode monolith.

An improved hydrophilic interaction/strong anion-exchange (HI-SAX) monolith for rapid capillary electrochromatographic profiling has been developed to detect carboxylic phytohormones with high resolution. The HI-SAX monolith was prepared by copolymerization of 2-(methacryloyloxy)ethyltrimethylammonium methyl sulfate, pentaerythritol triacrylate and porogenic solvents. Detailed polymerization compositions were investigated and improved, and the characteristics in terms of morphology, retention mechanism and column reproducibility were studied. A typical hydrophilic chromatography mechanism was observed when ACN content exceeded 30%, and the HI-SAX mixed-mode was verified with nucleic acid bases and nucleosides. Based on the improved HI-SAX monolith, capillary electrochromatographic profiling of typical phytohormones was evaluated by using indole-3-acetic acid (IAA), abscisic acid (ABA) and gibberellic acid (GA(3)) as the mode analytes. Effects of the test parameters on carboxyl phytohormones were investigated, and a fast analysis with high resolution for the representative phytohormones was achieved within 4.0 min with intraday RSDs (n = 3) less than 2.6% and 6.3% for the retention time and peak area, respectively. When applied to spiked corn samples, mean recoveries between 82.3% and 95.4% were obtained.

A facile versatile polymeric monolith for multiple separations.

A hydrophilic versatile polymeric monolith with multiple retention mechanisms including hydrogen-bonding, π-π and electrostatic interactions, was developed by a simple one-step in situ polymerization. High mechanical stability and excellent separation capabilities of various nonpolar and polar analytes were successfully achieved and employed for multiple separations in mixed-mode chromatography.

Online highly selective recognition of domoic acid by an aptamer@MOFs affinity monolithic column coupled with HPLC for shellfish safety monitoring.

Enabling cost-effective safety monitoring of shellfish is an important measure for the healthy development of the coastal marine economy. Herein, a new aptamer@metal-organic framework (MOF)-functionalized affinity monolithic column was proposed and applied in selective in-tube solid-phase microextraction (IT-SPME) coupled with HPLC for the accurate recognition of domoic acid (DA) in shellfish. Using a surface engineering strategy, ZIF-8 MOF was grown in situ inside the poly(epoxy-MA-co-POSS-MA) hybrid monolith. A high BET surface area and abundant metal reactive sites of the MOF framework were obtained for anchoring massive aptamers with terminal-modified phosphate groups. Various characterizations, such as SEM, elemental mapping, XRD, and BET, were performed, and the affinity performance was also studied. The presence of a massive amount of aptamers with a super coverage density of 3140 µmol L-1 bound on ZIF-8 MOF activated a high-performance bionic-affinity interface, and perfect specificity was exhibited with little interference of tissue matrixes, thus assuring the highly selective capture of DA from the complex matrixes. Under the optimal conditions, DA toxins in shellfish were detected with the limit of detection (LOD) of 7.0 ng mL-1 (equivalent to 14.0 µg kg-1), representing a 5-28 fold enhancement in detection sensitivity over traditional SPE or MIP adsorbents reported previously. The recoveries of fortified mussel and clam samples were achieved as 91.8 ± 1.2%-94.1 ± 1.9% (n = 3) and 91.2 ± 1.1%-94.5 ± 3.6% (n = 3), respectively. This work sheds light on a cost-effective method for online selective IT-SPME and the accurate monitoring of DA toxins using an aptamer@MOF-mediated affinity monolith system coupled with the inexpensive HPLC-UV technique.

Bioinspired polydopamine-mediated metal-organic framework click-grafting aptamers functionalized fabric for highly-specific recognition of microcystin-leucine arginine.

Fabricating functional electrospun nanofiber coating for highly selective extraction of microcystin-LR (MC-LR) was of significant importance for water-safety monitoring. Herein, a novel MOF@aptamer functionalized nanofabric was presented via a facile and reliable strategy integrating polydopamine (PDA) mediation and thiol-ene chemistry and applied for specific recognition of the MC-LR model analyte. Using polydopamine (PDA) as the mediating layer, vinyl-UiO-66 MOF was grown in situ, followed by post-synthetic modification (PSM) of Zr4+ with vinyl phosphate and rapid UV-initiated click reaction of aptamers. Uniform deposition of Zr-based MOF (vinyl-UiO-66) on the nanofibers was directly produced, and the tedious co-electrospinning process was abandoned to prevent the aggregation and encapsulation of MOF. Via an efficient "thiol-ene" chemistry, massive thiol-terminated aptamers were grafted on MOF within one step under friendly conditions, rather than the time-consuming nanoparticle adsorption or unfriendly covalent chemical reactions. As a result, the robust MOF@aptamer-coated nano-fabrics were obtained, and a highly selective performance towards MC-LR was illustrated with a limit of detection (LOD) at 0.002 ng/mL, good precision (CV<8.3%), good repeatability (2.2∼6.0%) when coupled with LC-MS. Almost 1∼2 orders of magnitude higher detection sensitivity was exhibited than that of the common non-specific SPE/SPME fiber reported so far. Applied to water samples, the good matrix-resistance ability, and acceptable recovery yields were achieved with high specificity. This strategy might provide a rapid and friendly protocol to efficiently fabricate MOF@aptamer functionalized nano-fabrics through electrospinning and interfacial "thiol-ene" chemistry for highly-selective microextraction.

Dual-Responsive Turn-On T1 Imaging-Guided Mild Photothermia for Precise Apoptotic Cancer Therapy.

Apoptosis has gained increasing attention in cancer therapy as an intrinsic signaling pathway, which leads to minimal leakage of waste products from a dying cell to neighboring normal cells. Among various stimuli to trigger apoptosis, mild hyperthermia is attractive but confronts limitations of non-specific heating and acquired resistance from elevated expression of heat shock proteins. Here, a dual-stimulation activated turn-on T1 imaging-based nanoparticulate system (DAS) is developed for mild photothermia (≈43 °C)-mediated precise apoptotic cancer therapy. In the DAS, a superparamagnetic quencher (ferroferric oxide nanoparticles, Fe3 O4 NPs) and a paramagnetic enhancer (Gd-DOTA complexes) are connected via the N6-methyladenine (m6 A)-caged, Zn2+ -dependent DNAzyme molecular device. The substrate strand of the DNAzyme contains one segment of Gd-DOTA complex-labeled sequence and another one of HSP70 antisense oligonucleotide. When the DAS is taken up by cancer cells, overexpressed fat mass and obesity-associated protein (FTO) specifically demethylates the m6 A group, thereby activating DNAzymes to cleave the substrate strand and simultaneously releasing Gd-DOTA complex-labeled oligonucleotides. The restored T1 signal from the liberated Gd-DOTA complexes lights up the tumor to guide the location and time of deploying 808 nm laser irradiation. Afterward, locally generated mild photothermia works in concert with HSP70 antisense oligonucleotides to promote apoptosis of tumor cells. This highly integrated design provides an alternative strategy for mild hyperthermia-mediated precise apoptotic cancer therapy.