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1.
J Nanosci Nanotechnol ; 20(2): 1066-1073, 2020 Feb 01.
Artigo em Inglês | MEDLINE | ID: mdl-31383106

RESUMO

Novozym435-gold nanoparticles biohybrids (NAuB) were synthesized and characterized in this work, with subsequent investigations on their catalytic activities and stability in compared with the initial form of Novozym435. This is a new way to modify immobilized enzyme by physical means. The elemental analysis of NAuB showed that Au was also the major element except C and O. The results of TEM and EDX demonstrated that gold nanoparticles (AuNPs) could be formed in the protein molecular network structure or on the surface of protein in the pores of Novozym435. The optimal process parameters for synthesis NAuB: synthesis temperature of 35 °C, HAuCl4 concentration of 0.02 mg/mL, synthesis time of 24 h. Using the transesterification of starch with ferulic acid ethyl ester as the reaction model, preliminary results pointed to excellent stabilities, solvent tolerance and activities as compared with Novozym435. As for a isooctane:dimethyl sulfoxide system, transesterification of starch with ferulic acid ethyl ester was carried out at 6% (w/w) NAuB and 60 °C for 24 h, a maximum DS of 0.2650 was obtained.

2.
Ann Anat ; 227: 151418, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-31626903

RESUMO

The present study aims to provide anatomical evidence for clinical application of the medial sural artery perforator (MSAP) flap. The current study investigated the vascular anatomy of the flap, evaluated the postoperative appearance and function of the donor and recipient sites, and investigate the clinical value in reconstruction of oral cavity. Six lower limbs of Chinese adult cadavers were microsurgically dissected. The locations and courses of the medial sural artery perforators were identified and recorded, which provided an anatomical basis for clinical application. Then, 16 clinical cases employing this flap were evaluated, ranging from 3×4cm to 6×8cm, and were employed for defects in the oral cavity region. Sixteen clinical cases with intraoral soft tissue defects, which included four clinical cases with inner cheek defects, were successfully followed up for 10-47 months (24 months on average). The donor site function, contour of recipient site and oral function recovery were evaluated as acceptable or better in cases with intraoral soft tissue defect, which were further verifying the value of clinical application of MSAP in repairing oral cavity defects. Moreover, two typical clinical cases were described in detail. To conclude, the MSAP flap is a favorable choice for small- to medium-size defects based on minor donor site morbidity, satisfactory oral function recovery, perforator stability and adaptation of the pedicle for anastomosis in the oral cavity region.

3.
Artigo em Inglês | MEDLINE | ID: mdl-31792734

RESUMO

The highly conserved heat shock protein 70 (HSP70) contributes to survival at a cellular level and greatly enhances stress tolerance in many organisms. In this study, we isolate and characterize Cshsp702, which encodes an inducible form of HSP70 in the rice stem borer, Chilo suppressalis. Cshsp702 does not contain introns; the translational product is comprised of 629 amino acids with an isoelectric point of 5.69. Real-time quantitative PCR revealed that Cshsp702 was expressed at maximal levels in hemocytes and was minimally expressed in the midgut. Expression of Cshsp702 in response to a range of temperatures (-11 to 43 °C) indicated significant induction by extreme cold and hot temperatures, with maximum expression after 2 h at 42 °C. The induction of Cshsp702 in response to the endoparasite Cotesia chilonis was also studied; interestingly, Cshsp702 expression in C. suppressalis was significantly induced at 24 h and 5 days, which correspond to predicted times of C. chilonis feeding and growth, respectively. The potential induction of Cshsp702 as an inflammatory response due to parasitic stress is discussed. In conclusion, Cshsp702 is induced in response to both environmental and biotic stress and plays an important role in the physiological adaptation of C. suppressalis.

4.
Small ; : e1904529, 2019 Dec 03.
Artigo em Inglês | MEDLINE | ID: mdl-31793714

RESUMO

van der Waals (vdW) layered materials have rather weaker interlayer bonding than the intralayer bonding, therefore the exfoliation along the stacking direction enables the achievement of monolayer or few layers vdW materials with emerging novel physical properties and functionalities. The ferroelectricity in vdW materials recently attracts renewed interest for the potential use in high-density storage devices. With the thickness becoming thinner, the competition between the surface energy, depolarization field, and interfacial chemical bonds may give rise to the modification of ferroelectricity and crystalline structure, which has limited investigations. In this work, combining the piezoresponse force microscope scanning, contact resonance imaging, the existence of the intrinsic in-plane polarization in vdW ferroelectrics CuInP2 S6 single crystals is reported, whereas below a critical thickness between 90 and 100 nm, the in-plane polarization disappears. The Young's modulus also shows an abrupt stiffness at the critical thickness. Based on the density functional theory calculations, these behaviors are ascribed to a structural phase transition from monoclinic to trigonal structure, which is further verified by transmission electron microscope technique. These findings demonstrate the foundational importance of structural phase transition for enhancing the rich functionality and broad utility of vdW ferroelectrics.

5.
J Chromatogr A ; : 460723, 2019 Nov 18.
Artigo em Inglês | MEDLINE | ID: mdl-31796249

RESUMO

Quantitative determination of endogenous compounds in biological samples has still been challenged by the absence of authentic blank matrix. Alternative strategy of surrogate matrix for preparing reference samples are prevalent due to its low cost and high availability. However, the evaluation system of surrogate matrix is not perfect. Herein, a novel multifunctional isotopic standards based steroidomics strategy was developed. Isotope-labeled standards were used not only as internal standards but also for the evaluation the feasibility of surrogate matrix, which improved the accuracy of assessment and could provide a new prospect for the quantitative analysis of endogenous compounds. Based on this approach, a detailed optimization from LC separation, MS detection to extraction conditions for global steroids in the steroidogenesis was firstly carried out. Characteristics and regularities of steroids in LC-MS were summarized to make references for further targeted or untargeted steroidomics study. Then eighteen steroids were quantified with high accuracy and high sensitivity in plasma from four types of cancer patients and healthy subjects using 1% BSA in PBS as surrogate matrix. And multi-steroids indexes with the best discriminating ability for lung, colorectal, breast and gastric cancer in different genders were identified successfully with Student's t-test, PLS-DA and logistic regression- ROC curve analysis. Finally, efficient cancer screening workflow was established by integrating the amine submetabolomics and lipidomics data of our previous studies. Taken together, the integrated steroidomics strategy could shed a light on the guidance for further steroidome as well as other endogenous compounds analysis and may provide a powerful tool for cancer diagnosis.

6.
Aging (Albany NY) ; 11(22): 10610-10625, 2019 Nov 27.
Artigo em Inglês | MEDLINE | ID: mdl-31785145

RESUMO

Proper activation of Toll-like receptor (TLR)-mediated signaling and production of proinflammatory cytokines are critical for the initiation of innate immunity, while the specific mechanism maintaining inflammatory homeostasis remains mostly unknown. Here, we show that Ets2 is upregulated following LPS and VSV stimulation. Ets2 knockdown or knockout leads to increased IL-6, TNF-α, and IFN-ß production in macrophages. Consistently, Ets2-deficient mice show exacerbated inflammatory cytokine production and are more susceptible to CLP-induced sepsis. Mechanistically, Ets2 inhibits the LPS- and VSV-induced activation of ERK1/2, JNK, p38, and p65. Ets2 also binds to the promoter of IL-6 to inhibit transcription. Collectively, the results of the present study show the negative regulatory role of Ets2 in LPS- and VSV-induced inflammation through the suppression of MAPK/NF-κB signaling, direct binding to the IL-6 promoter and inhibition of transcription.

7.
Small ; : e1905363, 2019 Nov 13.
Artigo em Inglês | MEDLINE | ID: mdl-31721439

RESUMO

Single atom catalysts (SACs) are receiving increasing interests due to their high theoretical catalytic efficiency and intriguing physiochemical properties. However, most of the synthetic methodologies involve high-temperature treatment. This usually leads to limited control over the spatial distribution of metal sites and collapse of porous network that result in limited active site exposure. A strategy to construct SAC by using a covalent organic framework as the precursor is reported in this study. The as-prepared catalyst is mainly composed of standing carbon layers with the presence of edge-site hosted metal single atoms. Such structure configuration not only allows full site exposure but also endows the metal site with high intrinsic activity. With a trace amount of cobalt loading (0.17 wt%), the nanorice-shaped catalyst displays promising electrochemical activities toward catalyzing the oxygen reduction reaction in both alkaline and acidic medium. An ultrahigh mass activity of 838 A gCo -1 at 0.9 V is achieved in the acidic electrolyte. This work suggests a new route to design SACs based on covalent organic framework for energy storage and conversion devices.

8.
Nat Commun ; 10(1): 5013, 2019 Nov 01.
Artigo em Inglês | MEDLINE | ID: mdl-31676774

RESUMO

For atomically thin two-dimensional materials, interfacial effects may dominate the entire response of devices, because most of the atoms are in the interface/surface. Graphene/sapphire has great application in electronic devices and semiconductor thin-film growth, but the nature of this interface is largely unknown. Here we find that the sapphire surface has a strong interaction with some of the carbon atoms in graphene to form a C-O-Al configuration, indicating that the interface interaction is no longer a simple van der Waals interaction. In addition, the structural relaxation of sapphire near the interface is significantly suppressed and very different from that of a bare sapphire surface. Such an interfacial C-O-Al bond is formed during graphene growth at high temperature. Our study provides valuable insights into understanding the electronic structures of graphene on sapphire and remote control of epitaxy growth of thin films by using a graphene-sapphire substrate.

9.
Diabetes ; 2019 Nov 11.
Artigo em Inglês | MEDLINE | ID: mdl-31712319

RESUMO

The whitening and loss of brown adipose tissue (BAT) during obesity and aging promote metabolic disorders and related diseases. The imbalance of Ca2+ homeostasis accounts for the dysfunction and clearance of mitochondria during BAT whitening. Capsaicin, a dietary factor activating TRPV1, can inhibit obesity induced by high-fat diet (HFD), but whether capsaicin inhibits BAT loss and the underlying mechanism remain unclear. In this study, we determined that the inhibitory effects capsaicin on HFD-induced obesity and BAT whitening were dependent on the participation of SIRT3, a critical mitochondrial deacetylase. SIRT3 also mediated all the beneficial effects of capsaicin on alleviating ROS generation, elevating mitochondrial activity and restricting mitochondrial calcium overload induced by HFD. Mechanistically, SIRT3 inhibits mitochondrial calcium uniporter (MCU)-mediated mitochondrial calcium overload by reducing the H3K27ac level on MCU promoter in an AMPK-dependent manner. In addition, HFD also inhibits AMPK activity to reduce SIRT3 expression, which could be reversed by capsaicin. Capsaicin intervention also inhibited aging-induced BAT whitening through this mechanism. In conclusion, this study emphasizes a critical role of AMPK/SIRT3 pathway in the maintenance of BAT morphology and function, and suggests that intervention in this pathway may be an effective target for preventing obesity or age-related metabolic diseases.

10.
Hypertension ; : HYPERTENSIONAHA11913778, 2019 Nov 18.
Artigo em Inglês | MEDLINE | ID: mdl-31735085

RESUMO

Endothelial dysfunction is an early step to the progression of cardiovascular diseases in diabetes. Apart from their anti-diabetic action, DPP-4 (dipeptidyl peptidase-4) inhibitors also reduce cardiovascular events in diabetic patients. However, the underlying mechanism of the beneficial effect of DPP-4 inhibitor on endothelial function is still obscure. In this study, we intervened type 1 or 2 diabetic model mice with vildagliptin for 4 weeks and measured the vascular reactivity. We found that vildagliptin improved endothelium-dependent vasodilation in diabetic mice independent of GLP-1 (glucagonlike peptide-1), but this effect was blocked by a SIRT1 (Sirtuin 1) inhibitor, Ex527. Mechanistically, vildagliptin-activated Transient Receptor Potential Channel Vanilloid 4 (TRPV4) to promote extracellular calcium uptake in endothelial cells, which activated AMPK (AMP-activated protein kinase)/SIRT1 pathway to counteract hyperglycemia-induced endothelial reactive oxygen species generation and senescence. Vildagliptin directly binds to TRPV4 by forming a hydrogen bond, which is critical to vildagliptin-evoked endothelial calcium intake. Knockout or inhibition of TRPV4 erased the beneficial role of vildagliptin. In addition, activation of SIRT1 by SRT1720 improved endothelial function independent of TRPV4 and reduced TRPV4 transcription to maintain an appropriate calcium level. In summary, our findings prove that vildagliptin protects against hyperglycemia-induced endothelial dysfunction by activating TRPV4-meditaed Ca2+ uptake, which helps to re-understand the mechanism of DPP-4 inhibitors and expand the therapeutic scope.

11.
Nat Commun ; 10(1): 5161, 2019 Nov 14.
Artigo em Inglês | MEDLINE | ID: mdl-31727890

RESUMO

Purely organic room temperature phosphorescence (RTP) has attracted wide attention recently due to its various application potentials. However, ultralong RTP (URTP) with high efficiency is still rarely achieved. Herein, by dissolving 1,8-naphthalic anhydride in certain organic solid hosts, URTP with a lifetime of over 600 ms and overall quantum yield of over 20% is realized. Meanwhile, the URTP can also be achieved by mechanical excitation when the host is mechanoluminescent. Femtosecond transient absorption studies reveal that intersystem crossing of the host is accelerated substantially in the presence of a trace amount of 1,8-naphthalic anhydride. Accordingly, we propose that a cluster exciton spanning the host and guest forms as a transient state before the guest acts as an energy trap for the RTP state. The cluster exciton model proposed here is expected to help expand the varieties of purely organic URTP materials based on an advanced understanding of guest/host combinations.

12.
PLoS Pathog ; 15(11): e1008169, 2019 Nov 18.
Artigo em Inglês | MEDLINE | ID: mdl-31738790

RESUMO

The unfolded protein response (UPR) in the endoplasmic reticulum (ER) constitutes a critical component of host innate immunity against microbial infections. In this report, we show that porcine reproductive and respiratory syndrome virus (PRRSV) utilizes the UPR machinery for its own benefit. We provide evidence that the virus targets the UPR central regulator GRP78 for proteasomal degradation via a mechanism that requires viral glycoprotein GP2a, while both IRE1-XBP1s and PERK-eIF2α-ATF4 signaling branches of the UPR are turned on at early stage of infection. The activated effector XBP1s was found to enter the nucleus, but ATF4 was unexpectedly diverted to cytoplasmic viral replication complexes by means of nonstructural proteins nsp2/3 to promote viral RNA synthesis. RNAi knockdown of either ATF4 or XBP1s dramatically attenuated virus titers, while overexpression caused increases. These observations reveal attractive host targets (e.g., ATF4 and XBP1s) for antiviral drugs and have implications in vaccine development.

13.
Med Phys ; 2019 Nov 19.
Artigo em Inglês | MEDLINE | ID: mdl-31742714

RESUMO

PURPOSE: As a novel imaging modality, X-ray fluorescence computed tomography (XFCT) can provide distribution and concentration information of contrast agents containing high atomic number elements, such as iodine, gadolinium, barium, gold, and platinum. Since XFCT has a better sensitivity and detection limit of high-Z elements comparing with traditional and spectral CT, it becomes a powerful quantitative imaging tool for biological studies. The main problem of current XFCT imaging is its low emission and detection efficiency of X-ray fluorescence (XRF) photons. Increasing XRF photons generation by choosing a high atomic element as a contrast agent is essential to improve the imaging quality of XFCT. Gadolinium emits at least a few times more of XRF photons than gold under the same X-ray excitation condition, leading to a detection limit at a level of sub-mg/mL as the next generation of clinical imaging modality. However, most current XFCT studies have utilized gadolinium salt as the contrast agent, which could not accumulate in organs or tumors efficiently, making in vivo XFCT imaging quite difficult. In this study, we present NaGdF4 nanoparticles with ultra-small size as nanoprobes to test the feasibility for in vivo XFCT application for the first time. METHODS: NaGdF4 nanoparticles with different sizes (3-10 nm) were successfully synthesized via a co-precipitation process by controlling the reaction time at temperature of 290 °C. The morphology, crystal phase, chemical composition, and size of such NPs were further characterized with HR-TEM, XRD, and EDX. The abilities of XRF photons from different sizes of NPs were quantified by our customized XFCT imaging system. To access the in-vivo application of as-synthesized NPs, such hydrophobic NPs capped OA molecules were further modified with AEP via a ligand-exchange process and characterized with FT-IR. For in-vivo XFCT imaging, 0.1 mL of 30 mg/mL NPs were injected into nude mice via the tail vein. The Varian G-297 X-ray tube was set to 150 kV and 0.5 mA. The XRF photons were captured by a Kromek eV-3500 photon counting detector at each 8° for 10 sec. RESULT: The successfully synthesized NaGdF4 nanoparticles (3-10 nm) were monodispersity, highly uniform spherical morphology and hexagonal crystalline phases. No significant influence on XRF photons yields or XFCT imaging quality were found by varying the nanoparticles size. The XRF photons were 2.5 times more emitted from NaGdF4 nanoparticles (NPs) compared to gold nanoparticles, thereby leading to a better image quality. With the AEP surface modification, such NPs were readily adapted for use in vivo XFCT applications with monodispersity in aqueous solution and negligible cytotoxicity. With the tail-vein injection, the liver, spleen and lungs could be clearly imaged with XFCT at a sub-mg/mL level. CONCLUSION: Such NaGdF4 NPs, which were synthesized with co-precipitation process, were modified with AEP for in-vivo XFCT applications. With both the phantom and in-vivo experiments, such NPs were proved to be an appropriate probe for XFCT application with the detection limit at a sub-mg/mL level. In the future research, such NPs could be further functionalized with targeting molecules for early-phase cancer detection.

14.
ACS Appl Mater Interfaces ; 11(47): 44673-44681, 2019 Nov 27.
Artigo em Inglês | MEDLINE | ID: mdl-31690067

RESUMO

Extreme heat events are mainly responsible for weather-related human mortality due to climate change. However, there is a lack of outdoor thermal management for protecting people from extreme heat events. We present a novel infrared-radiation-enhanced nanofiber membrane (NFM) that has good infrared resonance absorption and selectively radiates thermal radiation of the human body through the atmosphere and into the cold outer space. The NFM comprises polyamide 6 (PA6) nanofibers and randomly distributed SiO2 submicron spheres and has sufficient air permeability and thermal-moisture comfortability because of its interconnect nanopores and micropores. We measure the sky radiative cooling performance under a clear sky, and PA6/SiO2 NFM produces temperatures that are about 0.4-1.7 °C lower than those of commercial textiles when covering dry and wet hands and temperatures 1.0-2.5 °C lower than the ambient temperature when thermal conduction and convection are isolated in a closed device. Our processed PA6/SiO2 NFM combines sky radiative cooling with thermal management of the human body very well, which will promote the development of radiative cooling textiles.

15.
Cells ; 8(11)2019 Nov 07.
Artigo em Inglês | MEDLINE | ID: mdl-31703479

RESUMO

It is known that many diseases are caused by mutations or abnormalities in microRNA (miRNA). The usual method to predict miRNA disease relationships is to build a high-quality similarity network of diseases and miRNAs. All unobserved associations are ranked by their similarity scores, such that a higher score indicates a greater probability of a potential connection. However, this approach does not utilize information within the network. Therefore, in this study, we propose a machine learning method, called STIM, which uses network topology information to predict disease-miRNA associations. In contrast to the conventional approach, STIM constructs features according to information on similarity and topology in networks and then uses a machine learning model to predict potential associations. To verify the reliability and accuracy of our method, we compared STIM to other classical algorithms. The results of fivefold cross validation demonstrated that STIM outperforms many existing methods, particularly in terms of the area under the curve. In addition, the top 30 candidate miRNAs recommended by STIM in a case study of lung neoplasm have been confirmed in previous experiments, which proved the validity of the method.

16.
Sci Adv ; 5(11): eaav4355, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31700996

RESUMO

Recently, several captivating topological structures of electric dipole moments (e.g., vortex, flux closure) have been reported in ferroelectrics with reduced size/dimensions. However, accurate polarization distribution of these topological ferroelectric structures has never been experimentally obtained. We precisely measure the polarization distribution of an individual ferroelectric vortex in PbTiO3/SrTiO3 superlattices at the subunit cell level by using the atomically resolved integrated differential phase contrast imaging in an aberration-corrected scanning transmission electron microscope. We find, in vortices, that out-of-plane polarization is larger than in-plane polarization, and that downward polarization is larger than upward polarization. The polarization magnitude is closely related to tetragonality. Moreover, the contribution of the Pb─O bond to total polarization is highly inhomogeneous in vortices. Our precise measurement at the subunit cell scale provides a sound foundation for mechanistic understanding of the structure and properties of a ferroelectric vortex and lattice-charge coupling phenomena in these topological ferroelectric structures.

17.
Artigo em Inglês | MEDLINE | ID: mdl-31761733

RESUMO

Recent attempts have been made to direct the sensory neurotisation of free superficial circumflex iliac artery (SCIA) flaps. However, donor nerves enabling sensory recovery are limited. We report our findings in fifteen patients who underwent distal limb defect reconstruction using a sensate SCIA flap, including the lateral cutaneous branch of the subcostal nerve (LCSN), between August 2017 and September 2018. The distance from the anterosuperior iliac spine to the point where the LCSN crossed the iliac crest ranged between 6.5 and 10 cm. The diameter ranged between 1.5 and 4.0 mm. The flap size ranged between 8 × 4 and 13 × 10 cm2. All of the flaps survived uneventfully. Tests of sensory modalities, including the Semmes-Weinstein (SW) touch, vibration, pinprick, temperature and static two-point discrimination (s2PD) tests were applied in nine regions of each flap post-operatively. One or more modalities were present in at least one region at 6 months post-operatively, and the recovery of vibration perception was more consistent than that of SW touch and pinprick perception. The recovery of s2PD was noted in 4 cases over a follow-up period of more than 12 months. A reliable sensate flap with the LCSN can be considered as an attractive option for the sensory reconstruction of distal limb defects of moderate size.

18.
J Craniofac Surg ; 2019 Nov 21.
Artigo em Inglês | MEDLINE | ID: mdl-31764555

RESUMO

Crainoplasty following decompressive craniotomy is widely applied clinically. Here, a 55-year-old male had clinical presentation of fever, headache, and ulceration at the anterior scalp of left ear, with 4 to 5 mL pus induction per day, where he accepted cranioplastic surgery 17 years ago using the material of medical silicone rubber. The results of experimental test and magnetic resonance imaging indicated a sign of infection. The authors reported a rarely delayed infection 17 years after cranioplastic surgery. The over long-term risks for the previously transplanted materials should be recognized.

19.
Adv Mater ; : e1903491, 2019 Nov 14.
Artigo em Inglês | MEDLINE | ID: mdl-31725182

RESUMO

Metal oxides, as one of the mostly abundant and widely utilized materials, are extensively investigated and applied in environmental remediation and protection, and in energy conversion and storage. Most of these diverse applications are the result of a large diversity of the electronic states of metal oxides. Noticeably, however, many metal oxides present obstacles for applications in catalysis, mainly due to the lack of efficient active sites with desired electronic states. Here, the fabrication of single-tungsten-atom-oxide (STAO) is demonstrated, in which the metal oxide's volume reaches its minimum as a unit cell. The catalytic mechanism in the STAO is determined by a new single-site physics mechanism, named as quasi-atom physics. The photogenerated electron transfer process is enabled by an electron in the spin-up channel excited from the highest occupied molecular orbital to the lowest unoccupied molecular orbital +1 state, which can only occur in STAO with W5+ . STAO results in a record-high and stable sunlight photocatalytic degradation rate of 0.24 s-1 , which exceeds the rates of available photocatalysts by two orders of magnitude. The fabrication of STAO and its unique quasi-atom photocatalytic mechanism lays new ground for achieving novel physical and chemical properties using single-metal-atom oxides (SMAO).

20.
Anal Chem ; 2019 Dec 03.
Artigo em Inglês | MEDLINE | ID: mdl-31747259

RESUMO

Owing to the sharp edge-enhanced electric field, gold triangular nanoplates (AuNPLs) exhibit strong localized surface plasmon resonance (LSPR) scattering and display as red spots with high intensity and narrow distribution under dark-field microscopy (DFM), based on which sensitive sensing applications could be developed. As a concept of proof, the inhibition effects of pyrophosphate (PPi) against the etching of AuNPLs based on Cu2+ and I- mediated is dynamically monitored. The etching proceeding of AuNPLs by copper ions and iodide ions leads to the gradually blue-shifted LSPR scattering color of AuNPLs under DFM from the original red to green, accompanied by the reduced LSPR scattering intensity. As a result of the strong affinity of PPi to copper ions, the presence of PPi makes the etching process greatly suppressed, thereby achieving sensitive detection of the PPi.

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