CN115778381A - Intelligent closed-loop management system for in-situ detection and degradation of uric acid - Google Patents

Abstract

The invention relates to the field of medical instruments, and discloses an intelligent closed-loop management system for in-situ detection and degradation of uric acid. Meanwhile, the medicine can be used as the medicine basis of the existing treatment scheme for realizing the degradation of the whole body uric acid through the medicine, provides more accurate guidance for the safety range of the human blood uric acid, and can greatly enhance the convenience.

Description

An intelligent closed-loop management system for in situ detection and degradation of uric acid

technical field

The invention relates to the field of medical devices, in particular to an intelligent closed-loop management system for in-situ detection and degradation of uric acid.

Background technique

Hyperuricemia is the basis of gout. It is a disease in which purine metabolism disorder or uric acid excretion disorder leads to elevated blood uric acid. It is defined as a serum uric acid concentration greater than 7 mg/dL (420 μM) in men and greater than 6 mg/dL in women ( 360 μM). After a patient suffers from gout, monosodium urate (MSU) crystals tend to form in joint synovial fluid, first metatarsal, finger joints, knee joints, elbow joints, etc. Gouty arthritis is characterized by recurrent acute inflammation, Chronic tophi formation and bone erosion are the main clinical manifestations. Gout can be complicated by kidney disease. In severe cases, joint damage and renal function damage will occur. It is often accompanied by hyperlipidemia, hypertension, diabetes, arteriosclerosis and other diseases. All parts of the patient's body, especially the limbs, are prone to form tophi, which not only seriously affects the shape of the limbs, but can even lead to symptoms such as joint deformities and nerve compression. By strictly controlling the diet, blood uric acid can be reduced by 10-18%. When the effect of life intervention is not good, many gout patients take drugs (febuxostat and allopurinol, etc.) to control the progression of the disease. Uric acid level, and urate plays an anti-oxidation, blood pressure maintenance and boosting role in the body, and its concentration is too low (less than 180 μM) will affect the normal physiological state of people; in addition, for patients with different blood uric acid concentrations, drugs The dosage cannot be customized individually, it is easy to lead to abuse and cause side effects, and it cannot solve the problem from the root. When the diameter of tophi is greater than 1.5 cm, the patient’s epidermis is prone to ulceration, forming ulcers or sinus tracts that will not heal for a long time, which brings great inconvenience to daily life. The pain can only be relieved by surgery. Many middle-aged and elderly patients are not satisfied with surgery. Acceptance is not high, resulting in patients being affected by gout for many years.

Based on this, there is an urgent need to develop a device system for precise treatment of blood uric acid concentration in patients with hyperuricemia and gout. This system can not only reduce the patient's blood uric acid and eliminate gout symptoms by avoiding drugs and surgery. , while providing real-time monitoring (POCT) of blood uric acid concentration so that it is not lower than the safe value (180 μM), combined with mobile phone APP, to achieve closed-loop management of intelligent treatment for patients with hyperuricemia.

Biosensors are widely used in the detection of small molecule analytes such as glucose, uric acid, dopamine, and cholesterol in the body, mainly relying on the ability to selectively recognize biochemical substances and convert their concentrations into optical/electrical signals. A class of devices. This type of device has the advantages of high selectivity, low detection limit, high sensitivity, low production cost, multi-throughput screening, miniaturization, batch production and portability, and is widely used in biomedicine, food safety, Environmental testing and other fields. The biosensing detection system mainly includes four parts in its structure, namely, the detected analyte, the biosensitive recognition part (receptor), the signal conversion part (transducer) and the signal processing and output part. According to the working principle of the sensor, the receptor is very critical. In order to meet the needs of personalized detection in different sensing fields, new polymers, organic or inorganic nano Materials are introduced to improve detection performance, increase stability, and reduce manufacturing costs, among others. Among all kinds of biosensing applications, electrochemical biosensors are the most widely used ones because of their advantages of simplicity, speed, low cost, and diverse signal forms.

However, the current uric acid sensors only have the function of detection, and do not have the function of uric acid degradation.

Contents of the invention

In view of the limitations of the existing solutions for degrading uric acid in the whole body through drugs or removing tophi through surgery, the present invention provides an intelligent closed-loop management system for in-situ detection and degradation of uric acid. The intelligent closed-loop management system of the present invention can be used for the management of gout In-situ detection and treatment, especially the invasive in-situ electrochemical degradation of the joint cavity where tophi is prone to occur, is another option for the existing treatment plan to achieve the degradation of uric acid in the whole body through drugs or to remove tophi through surgery. Alternative options are also available for patients who are not candidates for surgery. At the same time, it can also be used as the basis for the existing treatment plan for the degradation of systemic uric acid through drugs, providing more accurate guidance for the safe range of human blood uric acid, and greatly enhancing the convenience.

Concrete technical scheme of the present invention is:

In the first aspect, the present invention provides an intelligent closed-loop management system for in-situ detection and degradation of uric acid based on a three-electrode system, including:

At least one uric acid sensitive electrode as the detection working electrode;

At least one uric acid-sensitive electrode as a working electrode for electrolysis;

reference electrode;

Electrode;

Electrochemical working terminals electrically connected to all electrodes.

Preferably, the reference electrode is an all-solid Ag/AgCl electrode based on carbon fiber; the counter electrode is a Pt wire electrode.

In the second aspect, the present invention provides an intelligent closed-loop management system for in-situ detection and degradation of uric acid based on a two-electrode system, including:

At least one uric acid sensitive electrode as the detection working electrode;

At least one uric acid-sensitive electrode as a working electrode for electrolysis;

A uric acid sensitive electrode as a counter electrode;

Electrochemical working terminals electrically connected to all electrodes.

Taking two intelligent closed-loop management systems based on the three-electrode system as an example, the working principle of the system of the present invention is: connect the detection/electrolysis working electrode, the reference electrode with the counter electrode and the electrochemical working terminal, and immerse all electrodes in saturated uric acid In the solution, the sensitive electrode is energized by the current-time method, and the current-uric acid concentration relationship curve is established. After calibrating the curve, slowly inject the working electrode, reference electrode and counter electrode into the body with a syringe containing physiological saline, Invade the growth site of the patient's tophi, still use the current-time method to energize, and transmit the read current data to the external mobile terminal (such as a smart phone terminal) through the electrochemical working terminal, according to the calibrated current-concentration curve Judging the current signal to determine normal operation or stop operation. When the input current is higher than the set value (for example, the blood uric acid concentration is higher than 420 μM), the electrochemical terminal is controlled to run the electrolysis working electrode, and work according to the preset electrolysis parameters; the current signal input from the electrolysis to the detection working electrode is lower than the threshold ( For example, when the blood uric acid concentration is lower than 180 μM), the electrolysis working electrode stops running, and the detection working electrode continues to work intermittently.

Preferably, the electrochemical working terminal is Hangzhou Lingzhi Technology Co., Ltd. - Zhihe 02CM, Shanghai Chenhua Instrument Co., Ltd. - CHI1200C and other models.

Preferably, there are multiple electrolytic working electrodes. It is further preferred that the number of detection working electrodes is one and the number of electrolysis working electrodes is three.

Preferably, the ends of the plurality of electrolytic working electrodes are bonded into a bundle by using semi-solid polyorthoester. It is further preferred that semi-solid polyorthoesters are used to bond the ends of multiple electrolytic working electrodes into a bundle at 2-3 mm.

During the experiment, the team of the present invention found that because the electrolytic working electrode needs to penetrate into the growth site of tophi, if the number is too large, multiple invasions are required, and the patient's compliance will be poor. For this reason, the present invention skillfully selects viscous polyorthoester as an adhesive to bond multiple electrolytic working electrodes into a bundle, which can reduce the number of invasions. Polyorthoesters are sensitive to acid, and the growth site of tophi is acidic due to the high concentration of uric acid. Therefore, polyorthoesters will degrade after intrusion, and the electrolytic working electrode can increase the electrolysis efficiency after being dispersed at the intrusion site. The decomposition products of polyorthoesters are water-soluble small molecules, which can be metabolized by organisms, have no biological toxicity, and have high safety.

As preferably, the uric acid sensitive electrode comprises:

Fibrous conductive substrate;

an ordered mesoporous carbon layer covering the surface of the fibrous conductive substrate;

A hydrogen peroxide catalyst loaded in pores of the ordered mesoporous carbon layer. It is used to catalyze the product hydrogen peroxide of the enzymatic reaction, and transfer the oxidized electrons of hydrogen peroxide to the ordered mesoporous carbon layer and the fibrous conductive substrate;

Uric acid oxidase loaded in pores of the ordered mesoporous carbon layer. It is used to selectively catalyze the substrate as hydrogen peroxide, so as to convert the detection of various analytes into the detection of hydrogen peroxide.

The advantage of the ordered mesoporous carbon layer of the present invention is that in addition to its good electrical conductivity, it can also significantly increase the specific surface area of the electrode, so that the electrode can load more catalysts; for supporting enzymes, the ordered mesoporous carbon also provides enzyme The confined space of the load makes the enzyme load more stable.

Preferably, the pore diameter of the ordered mesoporous carbon layer is 5-20 nm; the particle size of the hydrogen peroxide catalyst is less than 5 nm; the size of the urate oxidase is less than 20 nm.

Furthermore, the team of the present invention found through research that the ordered mesoporous carbon layer with the above-mentioned pore size can not only realize the transfer of electrons from the hydrogen peroxide catalyst to the conductive substrate, but also realize the confinement of the hydrogen peroxide catalyst and urate oxidase. load. If the pore size is too small, the hydrogen peroxide catalyst and urate oxidase cannot be loaded; if the pore size is too large, multiple enzyme molecules will be adsorbed in the pores, reducing the catalytic efficiency.

Preferably, the fibrous conductive substrate is selected from carbon fiber electrodes and carbon nanotube fiber electrodes.

Preferably, the hydrogen peroxide catalyst is selected from noble metal nanoparticles.

Preferably, the ordered mesoporous carbon layer is formed by in-situ growth of ordered mesoporous carbon material on the surface of the fibrous conductive substrate through hydrothermal reaction and then calcined at high temperature.

Further, the preparation method of the uric acid sensitive electrode comprises the following steps:

Step 1, modifying the ordered mesoporous carbon layer on the surface of the fibrous conductive substrate;

Step 2, loading a hydrogen peroxide catalyst on the ordered mesoporous carbon layer;

Step 3, continue to load urate oxidase on the surface of the ordered mesoporous carbon layer loaded with hydrogen peroxide catalyst; the loading method of urate oxidase is to utilize the confinement effect physical adsorption of the ordered mesoporous carbon layer or urate oxidase and organic Chemisorption between ordered mesoporous carbon layers.

Furthermore, in step 1, the single micelles composed of small organic molecules/surfactants are directly modified and grown on the surface of the fibrous conductive substrate through interfacial assembly, and an ordered mesoporous carbon layer is formed after calcination.

Further specifically, the preparation method of the uric acid sensitive electrode specifically includes the following steps:

Step A: preparing a low-order phenolic resin solution and a three-block copolymer solution respectively, and then mixing the low-order phenolic resin solution and the three-block copolymer solution, heating and stirring to obtain a spherical micelle solution;

Step B: taking the above-mentioned spherical micelle solution, diluting it with water, and at the same time immersing the fibrous conductive substrate in the obtained solution, and performing a hydrothermal reaction to obtain a fibrous conductive substrate with ordered micelles growing on the surface;

Step C: calcining the fibrous conductive substrate with ordered micelles grown on the surface in an inert gas atmosphere to obtain a fibrous conductive substrate with an ordered mesoporous carbon layer modified on the surface;

Step D: prepare the precursor solution of the hydrogen peroxide catalyst, immerse the fibrous conductive substrate with the ordered mesoporous carbon layer modified on the surface in the precursor solution, then add the reducing agent, after the reduction reaction, the peroxide-loaded Fibrous conductive substrates for hydrogen catalysts;

Step E: soak the fibrous conductive substrate loaded with the hydrogen peroxide catalyst in the urate oxidase solution, let it stand still, and take it out to prepare a uric acid sensitive electrode.

Preferably, in step C, the calcination temperature is 600-1200° C., and the calcination time is 2-6 hours.

Preferably, in step D, the precursor solution is an aqueous solution of chloroplatinic acid.

Compared with prior art, the beneficial effect of the present invention is:

(1) In the two intelligent closed-loop management systems based on three electrodes or two electrodes based on the three electrode system of the present invention, the degradation and detection of uric acid are respectively composed of multiple uric acid sensitive electrodes, a reference electrode and a counter electrode, and are combined with The electrochemical working terminal is connected, and the concentration of uric acid is read out in real time. This intelligent system is used for the in situ detection and treatment of gout, especially the invasive in situ electrochemical degradation of the joint cavity where tophi is prone to occur, as an existing treatment plan to achieve the degradation of systemic uric acid through drugs or remove it through surgery Another option for tophi, which also provides an alternative for patients who are not candidates for surgery. At the same time, it can also be used as the basis for the existing treatment plan for the degradation of systemic uric acid through drugs, providing more accurate guidance for the safe range of human blood uric acid, and greatly enhancing the convenience.

(2) The present invention is based on a fibrous fibrous conductive substrate, the surface of which is modified with an ordered mesoporous carbon layer, and further supports hydrogen peroxide catalyst and urate oxidase in the pores of the ordered mesoporous carbon layer to produce The obtained uric acid sensitive electrode not only has high conductivity, but also can significantly increase the specific surface area of the electrode, so that the electrode can load more catalysts; for the loaded enzyme, the ordered mesoporous carbon also provides a confined space for enzyme loading , making the enzyme load more stable, high sensitivity and high selectivity better.

(3) In order to further improve the convenience of use, the present invention uses polyorthoesters to bond multiple working electrodes into a bundle, which can reduce the number of invasions, and acid-sensitive polyorthoesters will occur when they enter areas with high uric acid concentrations. Degradation, the product has no biological toxicity, and the electrolysis efficiency can be increased after the working electrode is dispersed.

Description of drawings

The accompanying drawings show an exemplary embodiment of the present invention, and are used to explain the working principle of the present invention in conjunction with the description of the specific embodiment, including Fig. 1 to Fig. 7 to provide a further understanding of the present invention, and these drawings are included in this specification and form part of this manual.

Fig. 1 is the schematic diagram of the preparation method of a kind of uric acid sensitive electrode in the specific embodiment of the present invention;

Fig. 2 is the scanning electron micrograph of a kind of uric acid sensitive electrode in the specific embodiment of the present invention;

Fig. 3 is the construction flow diagram of a kind of intelligent closed-loop management system in the specific embodiment of the present invention;

Fig. 4 is the calibration curve of output current and uric acid concentration in the specific embodiment of the present invention;

Fig. 5 is the electrolysis curve of uric acid sensitive electrode to uric acid and tophi in the specific embodiment of the present invention;

Fig. 6 is the real tophi sample taken from the patient's body used in the specific embodiment of the present invention;

Fig. 7 is a schematic diagram of the comparison before and after the micro tophi is processed by the intelligent closed-loop management system in the specific embodiment of the present invention.

Detailed ways

The present invention will be further described below in conjunction with embodiment.

General Example

(1) An intelligent closed-loop management system for in-situ detection and degradation of uric acid based on a three-electrode system, including:

At least one uric acid sensitive electrode as the detection working electrode;

At least one uric acid-sensitive electrode as a working electrode for electrolysis;

Reference electrode (all solid-state Ag/AgCl electrode based on carbon fiber);

Counter electrode (Pt wire electrode);

The electrochemical working terminal (Hangzhou Lingzhi Technology Co., Ltd. - Zhibox 02CM, Shanghai Chenhua Instrument Co., Ltd. - CHI1200C) is electrically connected to all electrodes.

(2) An intelligent closed-loop management system for in-situ detection and degradation of uric acid based on a two-electrode system, including:

At least one uric acid sensitive electrode as the detection working electrode;

At least one uric acid-sensitive electrode as a working electrode for electrolysis;

A uric acid sensitive electrode as a counter electrode;

The electrochemical working terminal (Hangzhou Lingzhi Technology Co., Ltd. - Zhibox 02CM, Shanghai Chenhua Instrument Co., Ltd. - CHI1200C) is electrically connected to all electrodes.

In the above two intelligent closed-loop management systems, there are multiple electrolytic working electrodes. It is further preferred that the number of detection working electrodes is one and the number of electrolysis working electrodes is three.

As a preference, the ends (preferably at 2-3 mm) of the plurality of electrolytic working electrodes are bonded into a bundle by semi-solid polyorthoester.

Preferably, the uric acid sensitive electrode includes:

Fibrous conductive substrates (carbon fiber electrodes or carbon nanotube fiber electrodes);

An ordered mesoporous carbon layer covering the surface of the fibrous conductive substrate (formed by in-situ growth of ordered mesoporous carbon materials on the surface of the fibrous conductive substrate through hydrothermal reaction and then calcined at high temperature, with a pore size of 5-20 nm);

The hydrogen peroxide catalyst (noble metal nanoparticles, particle size less than 5 nm) loaded in the pores of the ordered mesoporous carbon layer is used to catalyze the product hydrogen peroxide of the enzymatic reaction, and transfer the electrons after hydrogen peroxide oxidation to On ordered mesoporous carbon layers and fibrous conductive substrates;

Uric acid oxidase (with a size less than 20 nm) loaded in the pores of the ordered mesoporous carbon layer. It is used to selectively catalyze the substrate as hydrogen peroxide, so as to convert the detection of various analytes into the detection of hydrogen peroxide.

Further, the preparation method of the uric acid sensitive electrode comprises the following steps:

Step 1, modifying the ordered mesoporous carbon layer on the surface of the fibrous conductive substrate (directly modifying and growing single micelles composed of small organic molecules/surfactants on the surface of the fibrous conductive substrate through interface assembly, and forming after calcination);

Step 2, loading a hydrogen peroxide catalyst on the ordered mesoporous carbon layer;

Step 3, continue to load urate oxidase on the surface of the ordered mesoporous carbon layer loaded with hydrogen peroxide catalyst; the loading method of urate oxidase is to utilize the confinement effect physical adsorption of the ordered mesoporous carbon layer or urate oxidase and organic Chemisorption between ordered mesoporous carbon layers.

Further specifically, the preparation method of the uric acid sensitive electrode specifically includes the following steps:

Step A: preparing a low-order phenolic resin solution and a three-block copolymer solution respectively, and then mixing the low-order phenolic resin solution and the three-block copolymer solution, heating and stirring to obtain a spherical micelle solution;

Step B: taking the above-mentioned spherical micelle solution, diluting it with water, and at the same time immersing the fibrous conductive substrate in the obtained solution, and performing a hydrothermal reaction to obtain a fibrous conductive substrate with ordered micelles growing on the surface;

Step C: calcining the fibrous conductive substrate with ordered micelles grown on the surface in an inert gas atmosphere (600-1200°C, 2-6 hours), to obtain a fibrous conductive substrate with an ordered mesoporous carbon layer modified on the surface;

Step D: prepare a precursor solution of hydrogen peroxide catalyst (preferably aqueous solution of chloroplatinic acid), immerse the fibrous conductive substrate with ordered mesoporous carbon layer on the surface in the precursor solution, and then add a reducing agent to reduce After the reaction, a fibrous conductive substrate loaded with a hydrogen peroxide catalyst is obtained;

Step E: soak the fibrous conductive substrate loaded with the hydrogen peroxide catalyst in the urate oxidase solution, let it stand still, and take it out to prepare a uric acid sensitive electrode.

Example 1

Step 1. Dissolve 0.5 g of phenol and 1.8 mL of 37% formaldehyde aqueous solution in 15 mL of 0.1mol/L sodium hydroxide aqueous solution, stir in a water bath at 67°C for 30 minutes, and obtain a light red solution;

Step 2. Dissolve 0.94 g triblock copolymer F127 in 65 mL of water to obtain a clear aqueous solution, then mix the light red low-order phenolic resin solution obtained in step 1 with the above triblock copolymer aqueous solution, and stir in a water bath at 67 °C for 10 Hour, obtain dark red spherical micelle solution;

Step 3: Take 9 g of the above-mentioned spherical micellar solution, add 20 g of water to dilute, and at the same time immerse carbon fibers less than 0.5 g in the obtained solution, and heat them in an oven at 130 ° C for 20 hours to obtain an orderly surface growth Micellar carbon fiber, take out the ordered mesoporous carbon fiber, wash and dry;

Step 4. Calcining the carbon fibers obtained in Step 3 at 600 °C for 2 hours in an inert gas atmosphere to obtain carbon fibers (with a pore size of about 10 nm) decorated with an ordered mesoporous carbon layer on the surface;

Step 5. Prepare 3 mL of 0.3 mmol/L aqueous solution of chloroplatinic acid, immerse the carbon fiber obtained in step 4 with a surface modified with an ordered mesoporous carbon layer in the above aqueous solution of chloroplatinic acid, and then dropwise add 160 μL of the mass fraction 0.07% sodium borohydride aqueous solution, put it in a refrigerator at 4°C for 8 hours to obtain ordered mesoporous carbon fibers loaded with platinum metal nanoparticles (average size 4.1 nm), take out ordered mesoporous carbon fibers, wash and dry ;

Step 6, prepare 160 μL of 1 mg/mL urate oxidase solution (the lattice parameter of the urate oxidase is 2.0 nm ( 1.8 nm ( 2.7 nm), and then immerse the ordered mesoporous carbon fiber obtained in step 5 in the above oxidase solution, put it in a refrigerator at 4°C for 8 hours, take it out, and dry it in a refrigerator at 4°C to obtain a fibrous uric acid sensitive electrode;

Fig. 1 shows the preparation process of above-mentioned uric acid sensitive electrode;

Figure 2 shows the TEM image of the fibrous uric acid sensitive electrode obtained according to the above preparation method, which shows the ordered mesoporous carbon layer on the surface of the carbon fiber, the loading of platinum metal nanoparticles on the surface of the ordered mesoporous carbon, and the uric acid Physisorption of oxidases on ordered mesoporous carbons;

Step 7. Cut the above-mentioned fibrous uric acid sensitive electrode into four sensitive electrodes of a certain length as the working electrode of the intelligent closed-loop control system. The reference electrode is an all-solid-state Ag/AgCl electrode based on carbon fiber, and the counter electrode is a Pt wire electrode. ;

Step 8. Connect the working electrode and reference electrode described in step 7 with the counter electrode and a small portable electrochemical working terminal (Shanghai Chenhua Instrument Co., Ltd. - CHI1200C), immerse all electrodes in 1mM saturated uric acid solution, and pass the current -Time method is energized to described sensitive electrode, establishes electric current-uric acid concentration relation curve, after calibrating curve, smear on working electrode end 2-3mm place with semi-solid polyorthoester to make it bond, described working electrode, The reference electrode and the counter electrode were slowly injected into the body with a syringe containing 300 μl of normal saline at a speed of 1.0 mm/s, and penetrated into the tophi growth site of the patient. The working electrodes that invaded the tophi growth site were divided into detection working electrodes and The electrolytic working electrode is equipped with 1 detection port and 3 electrolysis ports. Each port can apply voltage and read current. The applied voltage is 0-1 V, and the read current is 0.1-100 μA. The detection port adopts intermittent working mode. Detect once every 5 minutes, read the signal, and the electrolysis port continues to work; still use the current-time method to energize, transmit the read current data to the smartphone terminal, and judge the current signal transmitted to the mobile phone. According to the calibrated current- Concentration curve, when the input current is higher than the set value (the blood uric acid concentration is higher than 420 μM at this time), send a command to the portable electrochemical terminal, run the electrolysis port, and work according to the preset electrolysis parameters; electrolysis to the input of the detection port When the current signal is lower than the threshold (at this time, the blood uric acid concentration is lower than 180 μM), the command is sent again, the electrolysis port stops running, and the detection port continues to work intermittently;

Fig. 3 has shown the composition of intelligent closed-loop management system of the present invention and its running process;

Fig. 4 shows that in a specific embodiment, the current-concentration calibration curve of the sensitive electrode prepared according to the method of the present invention detects uric acid;

Figure 5 shows the electrochemical degradation effect on uric acid at different temperatures of the uric acid-sensitive electrode prepared according to the method of the present invention in a specific embodiment, and the electrolysis process of tiny tophi taken out of the patient's body;

Figure 6 shows the optical photograph of the tophus sample taken from the patient's body in a specific embodiment;

Figure 7 shows that in a specific embodiment, after being treated by the intelligent closed-loop management system for high blood uric acid and gout treatment, changes in tiny tophi appear, and the mass of tophi after electrolytic treatment drops from 1.3 mg to 0.7 mg ( Figure 7a) is before electrolysis, and Figure 7b) is after electrolysis), while the weight of the blank control sample (1.0mg) does not change significantly.

Example 2

Step 1. Dissolve 0.6 g of phenol and 2.1 mL of 37% formaldehyde aqueous solution in 15 mL of 0.1mol/L sodium hydroxide aqueous solution, and stir in a water bath at 68° C. for 30 minutes to obtain a light red solution;

Step 2: Dissolve 0.96 g of triblock copolymer F127 in 65 mL of water to obtain a clear aqueous solution, then mix the light red low-order phenolic resin solution obtained in Step 1 with the above triblock copolymer aqueous solution, and stir in a water bath at 68 °C for 12 Hour, obtain dark red spherical micelle solution;

Step 3: Take 10 g of the above-mentioned spherical micellar solution, add 20 g of water to dilute, and at the same time immerse carbon fibers less than 0.5 g in the obtained solution, and heat them in an oven at 130 ° C for 20 hours to obtain an orderly surface growth Micellar carbon fiber, take out the ordered mesoporous carbon fiber, wash and dry;

Step 4. Calcining the carbon fibers obtained in Step 3 at 800 °C for 4 hours in an inert gas atmosphere to obtain carbon fibers with an ordered mesoporous carbon layer on the surface (the pore size is about 15 nm);

Step 5. Prepare 3 mL of 0.3 mmol/L aqueous solution of chloroplatinic acid, immerse the carbon fiber obtained in step 4 with a surface modified with an ordered mesoporous carbon layer in the aqueous solution of chloroplatinic acid, and then dropwise add 180 μL of the mass fraction 0.075% sodium borohydride aqueous solution, put it in a refrigerator at 4°C for 8 hours to obtain ordered mesoporous carbon fibers loaded with platinum metal nanoparticles (average size 3.9 nm), take out the ordered mesoporous carbon fibers, wash and dry ;

Step 6, prepare 180 μL of 1 mg/mL urate oxidase solution (the lattice parameter of the urate oxidase is 2.0 nm ( 1.8 nm ( 2.7 nm), and then immerse the ordered mesoporous carbon fiber obtained in step 5 in the above oxidase solution, put it in a refrigerator at 4°C for 8 hours, take it out, and dry it in a refrigerator at 4°C to obtain a fibrous uric acid sensitive electrode;

Step 7. Cut the above-mentioned fibrous uric acid sensitive electrode into four sensitive electrodes of a certain length as the working electrode of the intelligent closed-loop control system. The reference electrode is an all-solid-state Ag/AgCl electrode based on carbon fiber, and the counter electrode is a Pt wire electrode. ;

Step 8. Connect the working electrode and reference electrode described in step 7 with the counter electrode and a small portable electrochemical working terminal (Hangzhou LinkZill Technology Co., Ltd. - Zhibox 02CM), and immerse the electrode in 1mM saturated uric acid solution , the sensitive electrode is energized by the current-time method, and the current-uric acid concentration relationship curve is established. After the calibration curve, the semi-solid polyorthoester is applied to the end of the working electrode at 2-3mm to make it bond, and the Using a syringe containing 300 μl of normal saline, the working electrode, reference electrode and counter electrode were slowly injected into the body at a speed of 1.0 mm/s, invading into the tophi growth site of the patient, and the working electrode that invaded the tophi growth site was divided into detection The working electrode and the electrolytic working electrode are equipped with 1 detection port and 3 electrolysis ports. Each port can apply voltage and read current. The applied voltage is 0-1 V, and the read current is 0.1-100 μA. The detection port adopts intermittent In the working mode, the detection is performed every 5 minutes, the signal is read, and the electrolytic port works continuously; the current-time method is still used to energize, and the read current data is transmitted to the smartphone terminal, and the current signal transmitted to the mobile phone is judged. The current-concentration curve, when the input current is higher than the set value (the blood uric acid concentration is higher than 420 μM at this time), send a command to the portable electrochemical terminal, run the electrolysis port, and work according to the preset electrolysis parameters; electrolysis to detection When the current signal input by the port is lower than the threshold (at this time, the blood uric acid concentration is lower than 180 μM), the command is sent again, the electrolysis port stops running, and the detection port continues to work intermittently.

Example 3

Step 1. Dissolve 0.7 g of phenol and 2.4 mL of 37% formaldehyde aqueous solution in 15 mL of 0.1mol/L sodium hydroxide aqueous solution, stir in a water bath at 70°C for 30 minutes, and obtain a light red solution;

Step 2. Dissolve 0.98 g triblock copolymer F127 in 65 mL of water to obtain a clear aqueous solution, then mix the light red low-order phenolic resin solution obtained in step 1 with the above triblock copolymer aqueous solution, and stir in a water bath at 70 °C for 14 Hour, obtain dark red spherical micelle solution;

Step 3: Take 11 g of the above-mentioned spherical micellar solution, add 20 g of water to dilute, and at the same time immerse carbon fibers less than 0.5 g in the obtained solution, and heat them in an oven at 130 ° C for 20 hours to obtain an orderly surface growth Micellar carbon fiber, take out the ordered mesoporous carbon fiber, wash and dry;

Step 4. Calcining the carbon fibers obtained in Step 3 at 1000 °C for 6 hours in an inert gas atmosphere to obtain carbon fibers with an ordered mesoporous carbon layer (pore diameter of about 18 nm) modified on the surface;

Step 5. Prepare 3 mL of 0.3 mmol/L chloroplatinic acid aqueous solution, immerse the carbon fiber obtained in step 4 with the surface modified with an ordered mesoporous carbon layer in the above chloroplatinic acid aqueous solution, and then dropwise add 200 μL of the mass fraction 0.08% sodium borohydride aqueous solution, put it in a refrigerator at 4°C for 8 hours to obtain ordered mesoporous carbon fibers loaded with platinum metal nanoparticles (average size 4.5 nm), take out ordered mesoporous carbon fibers, wash and dry ;

Step 6, prepare 200 μL of 1 mg/mL urate oxidase solution (the lattice parameter of the urate oxidase is 2.0 nm ( 1.8 nm ( 2.7 nm), and then immerse the ordered mesoporous carbon fiber obtained in step 5 in the above oxidase solution, put it in a refrigerator at 4°C for 8 hours, take it out, and dry it in a refrigerator at 4°C to obtain a fibrous uric acid sensitive electrode;

Step 7. Cut the above-mentioned fibrous uric acid sensitive electrode into four sensitive electrodes of a certain length as the working electrode of the intelligent closed-loop control system. The reference electrode is an all-solid-state Ag/AgCl electrode based on carbon fiber, and the counter electrode is a Pt wire electrode. ;

Step 8. Connect the working electrode and reference electrode described in step 7 with the counter electrode and a small portable electrochemical working terminal (Hangzhou LinkZill Technology Co., Ltd. - Zhibox 02CM), and immerse the electrode in 1mM saturated uric acid solution , the sensitive electrode is energized by the current-time method, and the current-uric acid concentration relationship curve is established. After the calibration curve, the semi-solid polyorthoester is applied to the end of the working electrode at 2-3mm to make it bond, and the Using a syringe containing 300 μl of normal saline, the working electrode, reference electrode and counter electrode were slowly injected into the body at a speed of 1.0 mm/s, invading into the tophi growth site of the patient, and the working electrode that invaded the tophi growth site was divided into detection The working electrode and the electrolytic working electrode are equipped with 1 detection port and 3 electrolysis ports. Each port can apply voltage and read current. The applied voltage is 0-1 V, and the read current is 0.1-100 μA. The detection port adopts intermittent In the working mode, the detection is performed every 5 minutes, the signal is read, and the electrolytic port works continuously; the current-time method is still used to energize, and the read current data is transmitted to the smartphone terminal, and the current signal transmitted to the mobile phone is judged. The current-concentration curve, when the input current is higher than the set value (the blood uric acid concentration is higher than 420 μM at this time), send a command to the portable electrochemical terminal, run the electrolysis port, and work according to the preset electrolysis parameters; electrolysis to detection When the current signal input by the port is lower than the threshold (at this time, the blood uric acid concentration is lower than 180 μM), the command is sent again, the electrolysis port stops running, and the detection port continues to work intermittently.

Raw materials used in the present invention, equipment, if not specified, are commonly used raw materials, equipment in this area; Method used in the present invention, if not specified, are conventional methods in this area.

The above are only preferred embodiments of the present invention, and do not limit the present invention in any way. All simple modifications, changes and equivalent transformations made to the above embodiments according to the technical essence of the present invention still belong to the technical solution of the present invention. scope of protection.

Claims (9)

1. An intelligent closed-loop management system for in-situ detection and uric acid degradation based on a three-electrode system is characterized by comprising:

at least one uric acid sensitive electrode as a detection working electrode;

at least one uric acid sensitive electrode as an electrolytic working electrode;

a reference electrode;

a counter electrode;

an electrochemical working terminal electrically connected to all of the electrodes.

2. The intelligent closed-loop management system of claim 1, wherein:

the reference electrode is an all-solid-state Ag/AgCl electrode based on carbon fibers; and/or

The counter electrode is a Pt wire electrode.

3. An intelligent closed-loop management system for in-situ detection and uric acid degradation based on a two-electrode system is characterized by comprising:

at least one uric acid sensitive electrode as a detection working electrode;

at least one uric acid sensitive electrode as an electrolytic working electrode;

a uric acid sensitive electrode as a counter electrode;

an electrochemical working terminal electrically connected to all the electrodes.

4. The intelligent closed-loop management system of one of claims 1-3, wherein: the number of the electrolytic working electrodes is multiple.

5. The intelligent closed-loop management system of claim 4, wherein: the tail ends of the plurality of electrolytic working electrodes are bonded into a bundle by adopting semi-solid polyorthoesters.

6. The intelligent closed-loop management system of one of claims 1-3, wherein: the uric acid sensitive electrode comprises: the conductive substrate comprises a fibrous conductive substrate, an ordered mesoporous carbon layer covering the surface of the fibrous conductive substrate, and a hydrogen peroxide catalyst and urate oxidase loaded in pores of the ordered mesoporous carbon layer.

7. The intelligent closed-loop management system of claim 6, wherein:

the aperture of the ordered mesoporous carbon layer is 5-20 nm; the particle size of the hydrogen peroxide catalyst is less than 5 nm; the size of the urate oxidase is less than 20 nm.

8. The intelligent closed-loop management system of claim 6, wherein: the preparation method of the uric acid sensitive electrode comprises the following steps: modifying an ordered mesoporous carbon layer on the surface of a fibrous conductive substrate; then loading a hydrogen peroxide catalyst and urate oxidase on the ordered mesoporous carbon layer in sequence.

9. The intelligent closed-loop management system of claim 8, wherein: the preparation method of the uric acid sensitive electrode specifically comprises the following steps:

step A: mixing the low-order phenolic resin solution with the triblock copolymer solution, heating and stirring to obtain a spherical micelle solution;

and B: b, adding water to dilute the product obtained in the step A, and then adding the product into a fibrous conductive substrate for hydrothermal reaction to obtain the fibrous conductive substrate with the ordered micelle growing on the surface;

and C: calcining the product obtained in the step B in an inert atmosphere to obtain the fibrous conductive substrate with the surface modified with the ordered mesoporous carbon layer;

step D: c, soaking the product obtained in the step C in a precursor solution of a hydrogen peroxide catalyst, and then carrying out reduction reaction to obtain a fibrous conductive substrate loaded with the hydrogen peroxide catalyst;

step E: and D, dipping the product obtained in the step D into a urate oxidase solution, standing and taking out to obtain the uric acid sensitive electrode.

Images