CN112345749A - Nucleic acid molecule embedded organic semiconductor small molecule aggregate, preparation method and application thereof in heavy metal ion detection - Google Patents

Nucleic acid molecule embedded organic semiconductor small molecule aggregate, preparation method and application thereof in heavy metal ion detection Download PDF

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CN112345749A
CN112345749A CN202011107646.1A CN202011107646A CN112345749A CN 112345749 A CN112345749 A CN 112345749A CN 202011107646 A CN202011107646 A CN 202011107646A CN 112345749 A CN112345749 A CN 112345749A
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nucleic acid
aggregate
organic semiconductor
alq3
acid molecules
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CN112345749B (en
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冯宇辉
崔春植
朴艳
崔龙吉
卜灿洙
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Yanbian University
Jilin Tobacco Industrial Co Ltd
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Jilin Tobacco Industrial Co Ltd
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    • GPHYSICS
    • G01MEASURING; TESTING
    • G01NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/5308Immunoassay; Biospecific binding assay; Materials therefor for analytes not provided for elsewhere, e.g. nucleic acids, uric acid, worms, mites
    • GPHYSICS
    • G01MEASURING; TESTING
    • G01NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N21/00Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
    • G01N21/62Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
    • G01N21/63Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
    • G01N21/64Fluorescence; Phosphorescence
    • G01N21/6428Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes"
    • GPHYSICS
    • G01MEASURING; TESTING
    • G01NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N21/00Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
    • G01N21/62Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
    • G01N21/63Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
    • G01N21/64Fluorescence; Phosphorescence
    • G01N2021/6417Spectrofluorimetric devices
    • YGENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y02TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02EREDUCTION OF GREENHOUSE GAS [GHG] EMISSIONS, RELATED TO ENERGY GENERATION, TRANSMISSION OR DISTRIBUTION
    • Y02E10/00Energy generation through renewable energy sources
    • Y02E10/50Photovoltaic [PV] energy
    • Y02E10/549Organic PV cells

Abstract

The invention discloses a nucleic acid molecule embedded organic semiconductor micromolecule aggregate, a preparation method and application thereof in heavy metal ion detection, belonging to the technical field of organic semiconductor micromolecule material application, wherein nucleic acid molecules can play a role of replacing a traditional surfactant in a crystalline organic semiconductor, namely Alq3 can be self-assembled to form an aggregate with a hexagonal prism shape under the guidance of single-stranded DNA. And DNA as a functional molecule capable of carrying information and the doping of Alq3 can play a role in enhancing the functionality of the material, and complementary DNA can play a role in enhancing the fluorescence of Alq3 by hybridizing on the surface of Alq 3. Compared with the surfactant, the DNA has the functions of transmitting and bearing information and specificity for identifying different substances. The breakthrough in the aspect of detection of the target object can be realized by utilizing the change of the nanostructure caused by the competitive complementation of the DNA sequence in the aggregate formed by self-assembly and the target object and the change of the optical signal caused by the change.

Description

Nucleic acid molecule embedded organic semiconductor small molecule aggregate, preparation method and application thereof in heavy metal ion detection
Technical Field
The invention belongs to the technical field of application of organic semiconductor micromolecule materials, and particularly relates to a nucleic acid molecule embedded organic semiconductor micromolecule aggregate, a preparation method and application thereof in heavy metal ion detection.
Background
Environmental pollution is a significant problem facing today's society. Heavy metals are highly toxic at trace levels, and their contamination poses a great threat to the environment and public health worldwide. The exploration of an efficient and sensitive method for detecting heavy metals is a necessary trend under high demand. At present, some more traditional detection and analysis means, such as inductively coupled plasma mass spectrometry, atomic absorption spectroscopy, ultraviolet-visible spectrophotometry, X-ray fluorescence spectroscopy and the like, are used for analyzing the heavy metal pollution degree of water bodies in large quantities. However, these methods usually rely on large-scale machines, the cost of instrument maintenance is high, and the sample preparation process is complicated during detection, so that the traditional detection technology has great limitations.
Aluminum octahydroxyquinoline (Alq3) was first mentioned in 1987 for the material preparation for organic light-emitting diodes. Since then, studies on Alq3 have focused on improving the brightness and long-term stability of Alq 3. Especially, the preparation of one-dimensional nanowires and nanorods has attracted people's attention. As in 2008, the Yadong Li project group first proposed that Alq3 micron rods having a hexagonal prism shape were prepared with the aid of a surfactant. Although the mechanism of self-assembly of the Alq3 bar in the surfactant solution is explained in the report, the research is stopped in the aspect of practical application.
In the molecular structure of DNA, two polydeoxyribonucleotide chains are coiled around a common central axis to form a double helix structure, and DNA is not only a natural biological information carrier, but also a structural material with multiple functions. Inspired by the unique physical and chemical properties of DNA, researchers use DNA to construct nano-objects with a reasonable architecture. For example, the unique base pairing rules and structural features of DNA can assemble plasmonic nanoparticles into structures with practical properties.
Disclosure of Invention
In order to overcome the defects that the detection method in the prior art usually depends on a large machine, the instrument maintenance cost is high, the sample preparation process is complicated during detection and the like, the invention provides the organic semiconductor micromolecule aggregate with the heavy metal ion detection function and the nucleic acid molecule embedded, wherein the nucleic acid molecule can replace the traditional surfactant in a crystalline organic semiconductor, namely, under the guidance of single-stranded DNA, Alq3 can be self-assembled to form the aggregate with the hexagonal prism shape. And DNA as a functional molecule capable of carrying information and the doping of Alq3 can play a role in enhancing the functionality of the material, and complementary DNA can play a role in enhancing the fluorescence of Alq3 by hybridizing on the surface of Alq 3. Compared with the surfactant, the DNA has the functions of transmitting and bearing information and specificity for identifying different substances. By means of the programmability of the DNA sequence, the flexibility of the structure design and the specific recognition effect of the specific DNA sequence on the specific target object, the nano-structure change caused by the competitive complementation of the DNA sequence in the aggregate formed by self-assembly and the target object and the change of the optical signal caused by the nano-structure change can be utilized, and the breakthrough in the aspect of detecting the target object can be realized.
The invention is realized by the following technical scheme:
an organic semiconductor small molecule aggregate with embedded nucleic acid molecules is an Alq3 rod-shaped aggregate which is synthesized by taking Alq3 powder as a material and using nucleic acid molecules (DNA) capable of specifically recognizing base sequences with specific heavy metal ions to assist self-assembly through biological guidance, and the nucleic acid molecules can be embedded in the Alq3 rod-shaped organic semiconductor aggregate in the self-assembly process.
Preferably, the specific heavy metal ion is a lead, cadmium, mercury or arsenic heavy metal ion.
Preferably, the sequence numbers of the nucleic acid molecules are respectively as follows:
5’-TTCTTTCTTCCCCTTGTTTGTT-3’、
5′-GGGTTCACAGTCCGTT-3′、
5'-ATGCAAACCCTTAAGAAAGTGGTCGTCCAAAAAACCATTG-3' or
5′-GGTTGGTGTGGTTGG-3′。
A preparation method of an organic semiconductor micromolecule aggregate with embedded nucleic acid molecules comprises the following specific steps:
taking nucleic acid molecule aqueous solutions with different base sequences and Alq3 powder solution dissolved by tetrahydrofuran, stirring, keeping out of the sun, and standing overnight at normal temperature to obtain an organic semiconductor micromolecule aggregate with embedded nucleic acid molecules; wherein the volume ratio of the nucleic acid molecule aqueous solution to the Alq3 powder solution is 8: 1.
Preferably, the concentration of the aqueous solution of nucleic acid molecules of different base sequences is 500nM single-stranded DNA per 16ml of solution.
Preferably, the sequence numbers of the nucleic acid molecules are respectively as follows:
5’-TTCTTTCTTCCCCTTGTTTGTT-3’、
5′-GGGTTCACAGTCCGTT-3′、
5'-ATGCAAACCCTTAAGAAAGTGGTCGTCCAAAAAACCATTG-3' or
5′-GGTTGGTGTGGTTGG-3′。
The invention also provides application of the nucleic acid molecule embedded organic semiconductor small molecule aggregate in detection of heavy metal ions.
Compared with the prior art, the invention has the following advantages:
the invention prepares the Alq3 micron rod in the hexagonal prism rod shape in the single-stranded DNA solution, and the nucleic acid molecule is embedded in the rod-shaped aggregate, and the Alq3 rod has stable fluorescence characteristics. By selecting different DNA chains, four different DNA sequences are selected, and the four heavy metal ions of mercury, cadmium, arsenic and lead are respectively identified in a specific manner. After the heavy metal ions are specifically identified, the fluorescence intensity of the Alq3 bar is obviously reduced. By observing the change of the fluorescence intensity of the Alq3 rod, the method can be used as a qualitative and quantitative means, and has sensitive response to heavy metal ions in a short time. Compared with the traditional detection means, the detection method is simpler and more convenient, consumes less time, has quick response and has more obvious detection effect.
Drawings
In order to more clearly illustrate the detailed description of the invention or the technical solutions in the prior art, the drawings that are needed in the detailed description of the invention or the prior art will be briefly described below. Throughout the drawings, like elements or portions are generally identified by like reference numerals. In the drawings, elements or portions are not necessarily drawn to scale.
FIG. 1 is a schematic representation of an aggregate of small organic semiconductor molecules into which nucleic acid molecules are embedded according to the present invention;
wherein, (a) is a scanning electron microscope, (b) is a high-resolution transmission microscope, and (c) is an energy dispersion spectrogram;
FIG. 2 is a graph comparing the effect of time on the change in fluorescence intensity of Alq3 bar solutions with and without added mercury ions;
wherein, (a) is a scanning electron micrograph, and (b) is a high-resolution transmission micrograph;
FIG. 3 is a fluorescence spectrum of chromium ion detection by a nucleic acid molecule-embedded organic semiconductor small molecule aggregate of the present invention;
FIG. 4 is a fluorescence spectrum of the nucleic acid molecule embedded organic semiconductor small molecule aggregate of the invention for arsenic ion detection;
FIG. 5 is a fluorescence spectrum of lead ion detection by the organic semiconductor small molecule aggregate embedded with nucleic acid molecules of the invention.
Detailed Description
The following embodiments are only used for illustrating the technical solutions of the present invention more clearly, and therefore, the following embodiments are only used as examples, and the protection scope of the present invention is not limited thereby.
It is to be noted that, unless otherwise specified, technical or scientific terms used herein shall have the ordinary meaning as understood by those skilled in the art to which the invention pertains.
Example 1
An organic semiconductor small molecule aggregate with embedded nucleic acid molecules is an Alq3 rod-shaped aggregate which is synthesized by taking Alq3 powder as a material and using nucleic acid molecules (DNA) capable of specifically recognizing base sequences with specific heavy metal ions to assist self-assembly through biological guidance, and the nucleic acid molecules can be embedded in the Alq3 rod-shaped organic semiconductor aggregate in the self-assembly process.
The specific heavy metal ions are lead, cadmium, mercury or arsenic heavy metal ions.
The sequence numbers of the nucleic acid molecules are respectively as follows:
5’-TTCTTTCTTCCCCTTGTTTGTT-3’、
5′-GGGTTCACAGTCCGTT-3′、
5'-ATGCAAACCCTTAAGAAAGTGGTCGTCCAAAAAACCATTG-3' or
5′-GGTTGGTGTGGTTGG-3′。
Self-assembly of Alq3 in a DNA solution can form aggregates with hexagonal prism morphology with fluorescence characteristics, and realize the embedding of DNA in the aggregates. After the heavy metal ion solution specifically recognized with DNA is added, the fluorescence intensity of Alq3 aggregate is obviously reduced, and the reduction amplitude is increased along with the increase of the concentration of the heavy metal. Thereby achieving the purpose of qualitative and quantitative analysis of heavy metal ions.
Example 2
A preparation method of a nucleic acid molecule embedded organic semiconductor micromolecule aggregate with a heavy metal ion detection function comprises the following specific steps: a single-stranded DNA (ss-DNA) aqueous solution of 16ml and 500nM each having a different base sequence was prepared, and 1mg/ml of Alq3 powder dissolved in tetrahydrofuran was added thereto, and the mixture was stirred for two minutes, protected from light, and left overnight at room temperature.
The sequence table of the single-stranded DNA with different base sequences is shown in Table 1.
Table 1 is a sequence listing of DNAs of four different base sequences
Figure BDA0002727498190000051
As shown in FIG. 1(a), it can be clearly seen from the scanning electron microscope that Alq3 successfully self-assembles in the single-stranded DNA solution to form a hexagonal prism rod-shaped structure with a smooth surface. And further confirmed by high resolution transmission microscope (HR-TEM), fig. 1(b) for morphology of solid hexagonal prism smooth rod type. To further confirm the self-assembly of Alq3 in the DNA solution, powder samples of Alq3 rods were coated on an ultra-thin porous copper mesh, and the presence of aluminum (Al) and phosphorus (P) elements, i.e., the composition of Alq3 molecules and ssDNA molecules, was confirmed according to the Energy Dispersive Spectroscopy (EDS) of fig. 1 (c).
To further demonstrate the stability of the Alq3 rod solution, a certain change in fluorescence intensity was observed at 60 minutes as shown in FIG. 2(a) when the Alq3 rod solution was irradiated with a 365nm UV lamp, which was caused by precipitation of the Alq3 rods in the solution during standing. After shaking, the fluorescence intensity of the solution without the addition of heavy metal ions is restored to the original value, the Alq3 solution maintains stable fluorescence characteristics, and the fluorescence intensity is reduced with the change of time after the addition of mercury ions. Further analysis of quantitative kinetic fluorescence detection using 365nm as excitation wavelength gave fluorescence intensity at 512 nm. As shown in FIG. 2(b), the fluorescence intensity of the well-dispersed Alq3 rod solution was measured by shaking sufficiently every one minute. The fluorescence intensity was significantly reduced after the addition of mercury ions, and as a comparison, the fluorescence intensity of the Alq3 rod solution without the addition of mercury ions remained relatively stable. According to the change of the fluorescence intensity with time, the kinetic equation can be fitted as follows:
I(t)=I0 exp(-kt) (1)
ln(I(t)/I0)=-kt (2)
wherein I (t) is the fluorescence intensity over time, I0K is the initial fluorescence intensity and is the constant of fluorescence decay. K without addition of heavy metal ionsbefore=0.010min-1Far less than k after adding heavy metal ionsafter=0.025min-1. It can be seen that the Alq3 rods are sensitive to heavy metal ions and respond quickly. It is further proved that the method is used for checkingFast and efficient detection of heavy metal ions.
Example 3
The invention also provides application of the nucleic acid molecule embedded organic semiconductor small molecule aggregate in detection of heavy metal ions.
Firstly, the method comprises the following steps: preparing 16ml of 500nM aqueous solution of single-stranded DNA (ss-DNA) which can be specifically identified with mercury ions, wherein the base sequence of the single-stranded DNA is as follows: 5'-TTCTTTCTTCCCCTTGTTTGTT-3' are provided. 2ml of 1mg/ml Alq3 powder dissolved in tetrahydrofuran was added, stirred for two minutes, protected from light and left overnight at room temperature.
And (3) mixing the prepared mercury ion solutions with the concentrations of 10, 100 and 200 mu g/ml respectively in a volume ratio of 9: 1 was mixed with the Alq3 rod prepared above. After 30min, the fluorescence intensities before and after the reaction were compared.
II, secondly: preparing 16ml of 500nM aqueous solution of single-stranded DNA (ss-DNA) which can be specifically identified with cadmium ions, wherein the base sequence of the single-stranded DNA is as follows: 5'-GGGTTCACAGTCCGTT-3' are provided. 2ml of 1mg/ml Alq3 powder dissolved in tetrahydrofuran was added thereto, and the mixture was stirred for two minutes, protected from light and left at room temperature overnight
Preparing prepared cadmium ion solutions with the concentrations of 10, 100 and 200 mu g/ml respectively according to the volume ratio of 9: 1 was mixed with the Alq3 rod prepared above. After 30min, the fluorescence intensities before and after the reaction were compared.
Thirdly, the method comprises the following steps: preparing 16ml of 500nM aqueous solution of single-stranded DNA (ss-DNA) which can be specifically identified with arsenic ions, wherein the base sequence of the single-stranded DNA is as follows: 5'-ATGCAAACCCTTAAGAAAGTGGTCGTCCAAAAAACCATTG-3' are provided. 2ml of 1mg/ml Alq3 powder dissolved in tetrahydrofuran was added thereto, and the mixture was stirred for two minutes, protected from light and left at room temperature overnight
And mixing the prepared arsenic ion solutions with the concentrations of 10, 100 and 200 mu g/ml respectively in a volume ratio of 9: 1 was mixed with the Alq3 rod prepared above. After 30min, the fluorescence intensities before and after the reaction were compared.
Fourthly, the method comprises the following steps: preparing 16ml of 500nM aqueous solution of single-stranded DNA (ss-DNA) which can be specifically identified with lead ions, wherein the base sequence of the single-stranded DNA is as follows: 5'-GGTTGGTGTGGTTGG-3' are provided. 2ml of 1mg/ml Alq3 powder dissolved in tetrahydrofuran was added thereto, and the mixture was stirred for two minutes, protected from light and left at room temperature overnight
Preparing a prepared lead ion solution with the concentration of 10, 100 and 200 mu g/ml respectively according to the volume ratio of 9: 1 was mixed with the Alq3 rod prepared above. After 30min, the fluorescence intensities before and after the reaction were compared.
When the method is adapted to other heavy metal ions such As cadmium (Cd), arsenic (As) and lead (Pb), the same good response effect is obtained. The fluorescence intensity of Alq3 decreased with the concentration of heavy metal ions.
As can be seen from fig. 3, 4 and 5, Alq3 had good fluorescence properties before the addition of heavy metal ions. An equal amount of deionized water was added as a control and fluorescence decreased within a reasonable range as Alq3 was diluted. After the heavy metal ions with different concentrations are added, the fluorescence is greatly reduced, and the reduction amplitude is increased along with the increase of the concentration of the heavy metal ions. Can achieve the purpose of qualitative analysis and preliminary quantitative analysis of heavy metal ions.
The preferred embodiments of the present invention have been described in detail with reference to the accompanying drawings, however, the present invention is not limited to the specific details of the above embodiments, and various simple modifications can be made to the technical solution of the present invention within the technical idea of the present invention, and these simple modifications are within the protective scope of the present invention.
It should be noted that the various technical features described in the above embodiments can be combined in any suitable manner without contradiction, and the invention is not described in any way for the possible combinations in order to avoid unnecessary repetition.
In addition, any combination of the various embodiments of the present invention is also possible, and the same should be considered as the disclosure of the present invention as long as it does not depart from the spirit of the present invention.
SEQUENCE LISTING
<110> Jilin tobacco industry, Limited liability company, Yanbian university
<120> organic semiconductor small molecule aggregate with embedded nucleic acid molecule, preparation method and application thereof in heavy metal ion
Applications of the detection aspect
<130> 2020.10.10
<160> 4
<170> PatentIn version 3.5
<210> 1
<211> 22
<212> DNA
<213> Artificial Synthesis
<400> 1
ttctttcttc cccttgtttg tt 22
<210> 2
<211> 16
<212> DNA
<213> Artificial Synthesis
<400> 2
gggttcacag tccgtt 16
<210> 3
<211> 40
<212> DNA
<213> Artificial Synthesis
<400> 3
atgcaaaccc ttaagaaagt ggtcgtccaa aaaaccattg 40
<210> 4
<211> 15
<212> DNA
<213> Artificial Synthesis
<400> 4
ggttggtgtg gttgg 15

Claims (7)

1. An organic semiconductor small molecule aggregate with embedded nucleic acid molecules is characterized in that the aggregate is an Alq3 rod-shaped aggregate which is synthesized by taking Alq3 powder as a material and using nucleic acid molecules (DNA) capable of specifically recognizing a base sequence with specific heavy metal ions to assist self-assembly through biological guidance, and the nucleic acid molecules can be embedded in the Alq3 rod-shaped organic semiconductor aggregate in the self-assembly process.
2. The aggregate of small organic semiconductor molecules into which nucleic acid molecules are inserted according to claim 1, wherein the specific heavy metal ion is a heavy metal ion selected from the group consisting of lead, cadmium, mercury, and arsenic.
3. The aggregate of small organic semiconductor molecules into which nucleic acid molecules are inserted according to claim 1, wherein the nucleic acid molecules have the following sequence numbers:
5’-TTCTTTCTTCCCCTTGTTTGTT-3’、
5′-GGGTTCACAGTCCGTT-3′、
5'-ATGCAAACCCTTAAGAAAGTGGTCGTCCAAAAAACCATTG-3' or
5′-GGTTGGTGTGGTTGG-3′。
4. The method for preparing the organic semiconductor small molecule aggregate embedded with the nucleic acid molecules according to claim 1, wherein the organic semiconductor small molecule aggregate embedded with the nucleic acid molecules is obtained by taking nucleic acid molecule aqueous solutions with different base sequences and Alq3 powder solution dissolved by tetrahydrofuran, stirring, keeping out of the sun, and standing overnight at normal temperature; wherein the volume ratio of the nucleic acid molecule aqueous solution to the Alq3 powder solution is 8: 1.
5. The method for producing an aggregate of small organic semiconductor molecules into which nucleic acid molecules are inserted according to claim 4, wherein the concentration of the aqueous solution of nucleic acid molecules having different base sequences is 500nM of single-stranded DNA per 16ml of the solution.
6. The method for preparing the aggregate of small organic semiconductor molecules with embedded nucleic acid molecules according to claim 4, wherein the sequence numbers of the nucleic acid molecules are respectively as follows:
5’-TTCTTTCTTCCCCTTGTTTGTT-3’、
5′-GGGTTCACAGTCCGTT-3′、
5'-ATGCAAACCCTTAAGAAAGTGGTCGTCCAAAAAACCATTG-3' or
5′-GGTTGGTGTGGTTGG-3′。
7. Use of the nucleic acid molecule-embedded organic semiconductor small molecule aggregate of claim 1 for heavy metal ion detection.
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