An intelligent diagnosis and treatment system and device for time-dependent insulin sensitivity impairment in the elderly based on Internet medical services

Abstract

The present invention discloses an intelligent diagnosis and treatment system and device for time-dependent insulin sensitivity impairment in the elderly based on Internet medical services. The present invention first reports the circadian rhythm of the key signaling molecule H2O2 in the liver redox system in elderly mice, subverting the classical theory that aging leads to oxidative stress in the past. It places the changes in the redox system during the aging process in the time dimension rather than only at the absolute quantity level, making the research on the imbalance of the redox system during the aging process more in-depth and effective. Based on this, the present invention provides an intelligent diagnosis and treatment system, device and computer-readable storage medium for insulin sensitivity impairment in the elderly based on Internet medical services, which has broad application prospects in the field of the treatment of insulin sensitivity impairment in the elderly and type II diabetes.

Description

Technical Field

[0001] The present invention belongs to the technical field of intelligent medicine, and particularly relates to an intelligent diagnosis and treatment system and device for time-dependent insulin sensitivity injury of the elderly based on Internet medical services. Background Art

[0002] During the aging process, metabolic disorders lead to the occurrence of age-related metabolic diseases such as type 2 diabetes. Type 2 diabetes is a chronic disease characterized by hyperglycemia caused by insulin resistance, making the body unable to fully respond to insulin. More and more studies have shown that oxidative stress plays a key role in the occurrence and progression of diabetes. The important signaling molecule H2O2 of the redox system regulates normal intracellular phosphorylation signals and the insulin signaling pathway by affecting the redox modification of key proteins such as IR, AKT, PTEN, and PTP1B. Therefore, using antioxidants to block the oxidation process in the body and avoid cell damage by free radicals is one of the current strategies for treating type 2 diabetes.

[0003] Existing studies have shown that endogenous H2O2 in the liver of young mice has a circadian rhythm oscillation. This redox signal rhythm participates in the regulation of the circadian rhythm and the regulation of the downstream signaling pathway of the biological clock through the redox modification of core clock proteins. Circadian rhythm disorder is one of the main characteristics of aging, and how the circadian rhythm of liver redox changes in aging individuals is still unknown. In recent years, the effect of using antioxidant drugs to treat type 2 diabetes in a large number of clinical trials has been minimal. In addition, a number of studies have shown that reactive oxygen species (ROS) can increase pro-survival signals, improve adaptability, and even extend the lifespan of yeast, worms, and mice, which is contradictory to the oxidative stress theory of aging. The relationship between the level of ROS and aging and its impact on elderly type 2 diabetes are still unclear.

[0004] It can be seen that the currently known regulatory mechanism of the redox system on glucose and lipid metabolism is not sufficient to support the application of antioxidant strategies to the clinical treatment of type 2 diabetes. The physiological regulation and pathological effects of reactive oxygen molecules on organisms not only lie in the change of the absolute amount, but also need to consider the change of rhythm. Therefore, exploring the change of the redox system rhythm during the aging process from the time dimension, and performing precise treatment with oxidants or antioxidants at specific times, specific sites, and specific targets are important scientific problems that need to be solved in the current clinical treatment of type 2 diabetes. Summary of the Invention

[0005] In view of this, in order to overcome the above technical problems existing in the current art, the purpose of the present invention is to provide an intelligent diagnosis and treatment system, device, and computer-readable storage medium for time-dependent insulin sensitivity injury of the elderly based on Internet medical services.

[0006] The present invention adopts the following technical solutions to achieve the above-mentioned invention objective:

[0007] In a first aspect of the present invention, there is provided an intelligent diagnosis and treatment system for elderly insulin sensitivity injury based on Internet medical services, and the system includes:

[0008] A diagnostic element information collector, configured to collect diagnostic element information related to the person to be diagnosed for elderly insulin sensitivity injury and send the collected data results to a disease diagnosis matcher;

[0009] A disease diagnosis matcher, configured to match the diagnostic element information of the diagnostic element information collector with disease types and send the matched data results to a treatment plan matcher;

[0010] A treatment plan matcher, configured to determine the result matched by the disease diagnosis matcher and match and call a treatment plan according to the result, and then send the treatment plan data results to a disease diagnosis and treatment information outputter;

[0011] A disease diagnosis and treatment information outputter, configured to output the data results received from the disease diagnosis matcher and the treatment plan matcher to a receiving unit;

[0012] The diagnostic elements for elderly insulin sensitivity injury include the expression level data of key proteins AKT and GSK3β in the liver insulin pathway, and the expression level data of liver insulin response proteins P-AKT and P-GSK3β;

[0013] In the disease diagnosis matcher, if the expression levels of key proteins AKT and GSK3β in the liver insulin pathway of the person to be diagnosed are significantly lower than those of young and healthy subjects, and the expression levels of liver insulin response proteins P-AKT and P-GSK3β are significantly lower than those of young and healthy subjects, then the person to be diagnosed is matched as a patient with elderly insulin sensitivity injury;

[0014] In the disease diagnosis matcher, if the expression levels of key proteins AKT and GSK3β in the liver insulin pathway of the person to be diagnosed have no significant difference from those of young and healthy subjects, and the expression levels of liver insulin response proteins P-AKT and P-GSK3β have no significant difference from those of young and healthy subjects, then the person to be diagnosed is matched as a non-elderly insulin sensitivity injury person;

[0015] In the treatment plan matcher, if the disease diagnosis matcher matches the person to be diagnosed as a patient with elderly insulin sensitivity injury, then the following treatment plan is matched and called: administering a therapeutically effective amount of buthionine sulfoximine to the person to be diagnosed during the period when H2O2 in the liver of the person to be diagnosed is reduced;

[0016] In the treatment plan matcher, if the disease diagnosis matcher matches the person to be diagnosed as a non-elderly insulin sensitivity impairment patient, there is no need to match and call the following treatment plan: administer a therapeutically effective amount of buthionine-sulfoximine to the person to be diagnosed during the period when the H2O2 in the liver of the person to be diagnosed decreases.

[0017] Further, the period when H2O2 decreases is ZT12-16.

[0018] Further, ZT12-16 is the time period from 12 hours to 16 hours after the start of light illumination.

[0019] Further, the concentration of buthionine-sulfoximine is 20 mM.

[0020] Further, the treatment plan can restore the insulin sensitivity of elderly insulin sensitivity impairment patients.

[0021] Further, the receiving unit is a display screen, a computer client, a mobile phone client, or a tablet.

[0022] The second aspect of the present invention provides an intelligent diagnosis and treatment device for elderly insulin sensitivity impairment based on Internet medical services. The device includes a memory and a processor; the memory is used to store program instructions; the processor is used to call the program instructions, and when the program instructions are executed, it is used to execute the following methods:

[0023] S101: Obtain data on diagnostic elements of elderly insulin sensitivity impairment related to the person to be diagnosed. The diagnostic elements of elderly insulin sensitivity impairment include the expression data of key proteins AKT and GSK3β in the liver insulin pathway, and the expression data of liver insulin response proteins P-AKT and P-GSK3β.

[0024] S102: Based on the data on diagnostic elements of elderly insulin sensitivity impairment related to the person to be diagnosed, determine whether the person to be diagnosed is an elderly insulin sensitivity impairment patient.

[0025] If the expression levels of key proteins AKT and GSK3β in the liver insulin pathway of the person to be diagnosed are significantly lower than those of young and healthy subjects, and the expression levels of liver insulin response proteins P-AKT and P-GSK3β are significantly lower than those of young and healthy subjects, then the person to be diagnosed is an elderly insulin sensitivity impairment patient.

[0026] S103: When the person to be diagnosed is an elderly insulin sensitivity impairment patient, select the following treatment plan: administer a therapeutically effective amount of buthionine-sulfoximine to the person to be diagnosed during the period when the H2O2 in the liver of the person to be diagnosed decreases.

[0027] Further, the period when H2O2 decreases is ZT12-16.

[0028] Furthermore, the concentration of buthionine-sulfoximine is 20 mM.

[0029] The third aspect of the present invention provides a computer-readable storage medium, on which a computer program is stored. When the computer program is executed by a processor, the following method is implemented:

[0030] S101: Obtain data on diagnostic elements for senile insulin sensitivity impairment related to the person to be diagnosed. The diagnostic elements for senile insulin sensitivity impairment include data on the expression levels of key proteins AKT and GSK3β in the hepatic insulin pathway, and data on the expression levels of hepatic insulin-responsive proteins P-AKT and P-GSK3β;

[0031] S102: Based on the data on diagnostic elements for senile insulin sensitivity impairment related to the person to be diagnosed, determine whether the person to be diagnosed is a patient with senile insulin sensitivity impairment;

[0032] If the expression levels of key proteins AKT and GSK3β in the hepatic insulin pathway of the person to be diagnosed are significantly lower than those of young and healthy subjects, and the expression levels of hepatic insulin-responsive proteins P-AKT and P-GSK3β are significantly lower than those of young and healthy subjects, then the person to be diagnosed is a patient with senile insulin sensitivity impairment;

[0033] S103: When the person to be diagnosed is a patient with senile insulin sensitivity impairment, select the following treatment plan: administer a therapeutically effective amount of buthionine-sulfoximine to the person to be diagnosed during the period when the hepatic H2O2 in the person to be diagnosed decreases.

[0034] Compared with the prior art, the present invention has the following advantages and beneficial effects:

[0035] (1) The present invention first reported the circadian rhythm of the key signal molecule H2O2 in the hepatic redox system in aged mice, subverting the classical theory that aging leads to oxidative stress in the past. It places the changes in the redox system during the aging process in the time dimension rather than only at the absolute quantity level, making the research on the imbalance of the redox system during the aging process more in-depth and effective. Based on this, the present invention provides an intelligent diagnosis and treatment system, device, and computer-readable storage medium for senile insulin sensitivity impairment based on Internet medical services, which has broad application prospects in the field of treatment of senile insulin sensitivity impairment and type II diabetes;

[0036] (2) By studying the changes in the redox system during the aging process from the time dimension, the present invention performs time-dependent oxidant treatment targeting the rhythm changes, thereby achieving the restoration of the rhythm of the key signaling molecule H2O2 in the redox system of the liver of old mice and the treatment of insulin sensitivity injury, providing a new solution for the failure of simple antioxidant treatment of type II diabetes in clinical practice at present, and will promote the development and application of precision treatment strategies for elderly type II diabetes, which is of great significance;

[0037] (3) The proposal of the present invention not only provides a new idea for solving the failure of antioxidant drug treatment of type II diabetes in clinical practice and the oxidative stress paradox during the aging process, that is, precisely intervening from the time dimension to restore the redox homeostasis of elderly individuals, but also the oxidant buthionine-sulfoximine (BSO) used in the technical solution provided by the present invention has been approved for anti-tumor treatment, with extremely high safety and reliability, and is expected to be used for the treatment of elderly type II diabetes. BRIEF DESCRIPTION OF THE DRAWINGS

[0038] Figure 1 : Schematic diagram of an intelligent diagnosis and treatment system for insulin sensitivity injury in the elderly based on Internet medical services provided by an embodiment of the present invention;

[0039] Figure 2 : Schematic diagram of an intelligent diagnosis and treatment device for insulin sensitivity injury in the elderly based on Internet medical services provided by an embodiment of the present invention;

[0040] Figure 3 : Corresponding result graph of the content detection of the circadian rhythm of H2O2 in the liver of mice;

[0041] Figure 4 : Corresponding result graph of the insulin sensitivity detection after time-dependent BSO drinking water in old mice;

[0042] Figure 5 : Corresponding result graph of the liver H2O2 content detection after time-dependent BSO drinking water in old mice;

[0043] Figure 6 : Corresponding result graph of the change in the insulin pathway in the liver after time-dependent BSO drinking water in old mice detected by Western blot. DETAILED DESCRIPTION OF THE EMBODIMENTS

[0044] The present invention will be further described below in conjunction with specific embodiments. The following specific embodiments are only used to explain the present invention and should not be construed as limiting the present invention. Those of ordinary skill in the art can understand that various changes, modifications, substitutions, and variations can be made to these embodiments without departing from the principles and purposes of the present invention. The scope of the present invention is defined by the claims and their equivalents. The reagents and raw materials used in the present invention are easily obtained by those of ordinary skill in the art and can be obtained from commercial sources without special instructions. The experimental methods without specific conditions mentioned in the present invention are usually carried out under conventional conditions or according to the conditions recommended by the manufacturer. In particular, the following embodiments are only used to illustrate the present invention and should not limit the scope of the present invention in any way. It should be noted that the experimental conditions and results described in the following embodiments are only used to illustrate the present invention and should not and will not limit the present invention described in detail in the claims.

[0045] In order to enable those skilled in the art to better understand the solution of the present invention, the technical solutions in the embodiments of the present invention will be clearly and completely described below in conjunction with the drawings in the embodiments of the present invention. In some processes described in the specification and claims of the present invention and the above drawings, a plurality of operations appear in a specific order. However, it should be clearly understood that these operations may not be executed in the order in which they appear in this article or may be executed in parallel. The serial numbers of the operations, such as S101, S102, S103, etc., are only used to distinguish different operations, and the serial numbers themselves do not represent any execution order. In addition, these processes may include more or fewer operations, and these operations may be executed in sequence or in parallel.

[0046] The technical solutions in the embodiments of the present invention will be clearly and completely described below in conjunction with the drawings in the embodiments of the present invention. Obviously, the described embodiments are only a part of the embodiments of the present invention, rather than all the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative work belong to the scope of protection of the present invention.

[0047] Figure 1 It is a schematic diagram of an intelligent diagnosis and treatment system for elderly insulin sensitivity injury based on Internet medical services provided by an embodiment of the present invention. The system includes:

[0048] A diagnostic element information collector, configured to collect diagnostic element information related to the person to be diagnosed for elderly insulin sensitivity injury and send the collected data results to a disease diagnosis matcher;

[0049] A disease diagnosis matcher, configured to match the diagnostic element information of the diagnostic element information collector with disease types and send the matched data results to a treatment plan matcher;

[0050] A treatment plan matcher, which is used to determine the result matched by the disease diagnosis matcher, match and call a treatment plan according to the result, and then send the treatment plan data result to the disease diagnosis and treatment information outputter;

[0051] A disease diagnosis and treatment information outputter, which is used to output the data results received from the disease diagnosis matcher and the treatment plan matcher to a receiving unit;

[0052] The diagnostic elements of elderly insulin sensitivity injury include the expression data of key proteins AKT and GSK3β in the hepatic insulin pathway, and the expression data of hepatic insulin-responsive proteins P-AKT and P-GSK3β;

[0053] In the disease diagnosis matcher, if the expression levels of key proteins AKT and GSK3β in the hepatic insulin pathway of the person to be diagnosed are significantly lower than those of young and healthy subjects, and the expression levels of hepatic insulin-responsive proteins P-AKT and P-GSK3β are significantly lower than those of young and healthy subjects, then the person to be diagnosed is matched as a patient with elderly insulin sensitivity injury;

[0054] In the disease diagnosis matcher, if the expression levels of key proteins AKT and GSK3β in the hepatic insulin pathway of the person to be diagnosed have no significant difference from those of young and healthy subjects, and the expression levels of hepatic insulin-responsive proteins P-AKT and P-GSK3β have no significant difference from those of young and healthy subjects, then the person to be diagnosed is matched as a non-elderly insulin sensitivity injury person;

[0055] In the treatment plan matcher, if the disease diagnosis matcher matches the person to be diagnosed as a patient with elderly insulin sensitivity injury, then the following treatment plan is matched and called: administering a therapeutically effective amount of buthionine sulfoximine to the person to be diagnosed during the period when H2O2 in the liver of the person to be diagnosed decreases;

[0056] In the treatment plan matcher, if the disease diagnosis matcher matches the person to be diagnosed as a non-elderly insulin sensitivity injury person, then there is no need to match and call the following treatment plan: administering a therapeutically effective amount of buthionine sulfoximine to the person to be diagnosed during the period when H2O2 in the liver of the person to be diagnosed decreases.

[0057] Further, the period when H2O2 decreases is ZT12 - 16.

[0058] Further, ZT12 - 16 is the time period from 12 hours to 16 hours after the start of light.

[0059] Further, the concentration of buthionine sulfoximine is 20 mM.

[0060] Further, the treatment plan can restore the insulin sensitivity of patients with elderly insulin sensitivity injury.

[0061] Further, the receiving unit is a display screen, a computer client, a mobile phone client or a tablet computer.

[0062] In some embodiments, the P-AKT refers to phosphorylated AKT (P-AKT), and the P-GSK3β refers to phosphorylated glycogen synthase kinase-3β (P-GSK3β).

[0063] In some embodiments, the buthionine sulfoximine refers to L-buthionine sulfoximine.

[0064] In a specific embodiment, the present invention detected the circadian rhythm content of H2O2 in the livers of mice, and for the first time found that the content of H2O2 in the livers of young mice has a circadian rhythm, which increases during the active period and reaches a peak at ZT18; while the content of H2O2 in the livers of old mice loses the circadian rhythm fluctuation and is continuously lower than that of young mice. The specific experimental materials, experimental methods and experimental results are as follows:

[0065] (1) Experimental materials

[0066] Hydrogen peroxide content detection kit: Red Hydrogen Peroxide / Peroxidase AssayKit (A22188, ThermoFisher);

[0067] Mouse synchronization: 3-month-old and 18-month-old C57 / 6J mice drink water and eat freely, and are synchronized in an environment with a 24-hour light-dark cycle. The lights are on from 9 am to 9 pm, and off from 9 pm to 9 am the next day, for more than 2 weeks.

[0068] (2) Experimental methods

[0069] ① Reagent preparation

[0070] Prepare 1×Reaction Buffer: Warm the 5×Reaction Buffer stock solution to room temperature, dilute it with deionized water to 1×Reaction Buffer, and vortex thoroughly.

[0071] Prepare the working solution: Warm the Red and HRP stock solutions to room temperature, gently flick the tube wall to mix, and add them to 1×Reaction Buffer, with the final concentration of Red being 100 μM and the final concentration of HRP being 0.2 U / mL. For example, when preparing 5 mL of the working solution, add 50 μL of Red stock solution and 100 μL of HRP stock solution to 4.85 mL of 1×Reaction Buffer. The working solution can be stably stored for 1 day.

[0072] ② Sample preparation

[0073] Take out the liver tissue from the -80°C refrigerator. Cut 15 mg of the liver and place it in a 2 mL round-bottom EP tube containing 600 μL of 1×Reaction Buffer. Add grinding beads to the tube, place it in a tissue grinder, grind at 60 Hz for 180 s, take out the grinding beads, centrifuge at 12,000 rpm at 4°C for 10 min, take the supernatant and transfer it to a new 1.5 mL EP tube, and place it on ice for later use.

[0074] ③ Prepare the standard curve

[0075] Prepare the H2O2 standard curve. First, dilute the H2O2 stock solution 1000-fold with 1×Reaction Buffer to a final concentration of 8.8 mM. Then, sequentially dilute the 8.8 mM H2O2 to 100, 50, 25, 12.5, 6.25, 1.25, 0 μM. The low-concentration H2O2 is unstable and needs to be prepared freshly before use.

[0076] ④ Determine H2O2

[0077] Preliminary experiment: Take one tube of the sample, dilute it 10 and 20 times, then take 50 μL and add it to a 96-well plate with a completely black bottom and walls. Add 50 μL of the working solution to each well and mix evenly. Incubate at room temperature in the dark for 30 min, and read the relative fluorescence units (RFUs) using a microplate reader. Determine the dilution factor of the experimental sample based on the fluorescence intensity of the sample in the preliminary experiment.

[0078] Formal experiment: According to the dilution factor obtained from the preliminary experiment, dilute each sample in a new 1.5 mL EP tube at a volume of two replicates, and then add it to a completely black 96-well plate together with the standard curve. Quickly add 50 μL of the working solution to each well in the dark, incubate at room temperature in the dark for 30 min, and read the relative fluorescence units (RFUs) using a microplate reader. The excitation wavelength is 545 nm and the emission wavelength is 590 nm. The corresponding H2O2 content in the sample is calculated according to the standard curve.

[0079] (3) Experimental results

[0080] The experimental results are as Figure 3 shown. The H2O2 content in the liver of young mice has a circadian rhythm, increasing during the active period and reaching a peak at ZT18; while the H2O2 content in the liver of old mice has lost the circadian rhythm fluctuation and is continuously lower than that of young mice.

[0081] In some embodiments, ZT represents "Zeitgeber Time", that is, "time signal", which is used to identify the circadian time in an organism. ZT18 refers to a time point used to describe circadian rhythm experiments in biological and medical research. In such a system, ZT0 represents the start of the light cycle (i.e., the start of the "light period"), while ZT12 represents the end of the light cycle (i.e., the start of the "dark period"). ZT18 represents a time point in the second half of the light cycle, that is, 18 hours after the start of light. ZT12-16 represents the time period from 12 hours to 16 hours after the start of light.

[0082] In a specific embodiment, the present invention detected the insulin sensitivity of old mice after time-dependent BSO water intake, and found that compared with young mice, the insulin tolerance of old mice was significantly decreased and the insulin sensitivity was damaged; while feeding BSO water (Aged-BSO-4h) during the active period ZT12-16 could partially restore the insulin sensitivity of old mice, and the blood glucose was significantly different from that of normal old mice at 15 and 30 minutes; while the insulin tolerance of old mice fed BSO water (Aged-BSO-24h) throughout the day was similar to that of normal old mice, and the decrease in insulin sensitivity caused by aging was not restored, indicating that only oxidant treatment during the period when the liver H2O2 of old mice was reduced (i.e., the active period ZT12-16) could restore the insulin sensitivity of old mice to a certain extent, while continuous oxidant treatment did not restore the insulin sensitivity of old mice. This first discovered result belongs to a technical effect unexpected by those skilled in the art based on the prior art. The specific experimental materials, experimental methods and experimental results are as follows:

[0083] (1) Experimental materials

[0084] L-Buthionine sulfoximine (L-BSO) (HY-106376A, MCE); insulin (91077C, Merck); blood glucose test strips (590, Yuyue).

[0085] (2) Experimental methods

[0086] ① Time-dependent BSO water intake for old mice

[0087] The synchronized old mice were randomly divided into three groups. The control group mice drank water freely for 24 hours; the mice in the BSO-4h group drank BSO water (20 mM) during the active period ZT12-16 and normal water at other times; the mice in the BSO-24h group drank BSO water (20 mM) for 24 hours, and the water intake continued for 2 weeks. Young mice drank water freely as a control.

[0088] ② Mouse insulin tolerance test

[0089] After two weeks of BSO water feeding, young and old mice were fasted for 6 hours (ZT12 - 18). At ZT18, they were weighed, and the volume of insulin injection solution needed for each mouse was calculated (0.75 IU insulin / kg body weight). First, the fasting blood glucose level was measured: The tip of the tail was cut with sterilized surgical scissors, the first drop of blood was discarded, fresh blood was squeezed out, dropped on the blood glucose test strip, and the reading was recorded. Then, according to the calculated injection volume, all mice were injected with insulin intraperitoneally, the injection time was recorded, and blood glucose was detected 5 times at 15, 30, 45, 60, and 120 minutes after injection. Before each detection, the scab at the tail tip wound was removed, the tail was massaged, and fresh blood drops were taken for detection.

[0090] (3) Experimental results

[0091] The experimental results are as Figure 4 shown. Compared with young mice, the insulin tolerance of old mice decreased significantly, and insulin sensitivity was damaged; while feeding BSO water during the active period ZT12 - 16 (Aged - BSO - 4h) could partially restore the insulin sensitivity of old mice, and the blood glucose was significantly different from that of normal old mice at 15 and 30 minutes; while the insulin tolerance of old mice fed BSO water throughout the day (Aged - BSO - 24h) was similar to that of normal old mice, and the decrease in insulin sensitivity caused by aging was not restored. It shows that only by performing oxidant treatment during the period when hepatic H2O2 in old mice decreases, that is, during the active period ZT12 - 16, can the insulin sensitivity of old mice be restored to a certain extent, while continuous oxidant treatment does not restore the insulin sensitivity of old mice.

[0092] In some embodiments, the BSO refers to Buthionine sulfoximine (BSO). BSO is an irreversible inhibitor of γ - glutamylcysteine synthetase, which can reduce the level of glutathione in tissue cells. Currently, BSO has been applied as a tumor chemotherapy sensitizer to enhance the effect of radiotherapy and in combination with other drugs to improve the cancer treatment effect. In the specific implementation of the present invention, the BSO refers to L - Buthionine sulfoximine (L - BSO).

[0093] In a specific embodiment, the present invention detected the content of H2O2 in the liver of aged mice after time-dependent BSO drinking water. It was found that after two weeks of BSO drinking water, at ZT18, the content of H2O2 in the liver of aged mice in the Aged-BSO-4h group was significantly increased compared with that of normal aged mice, slightly lower than that of young mice; while the content of H2O2 in the liver of aged mice in the Aged-BSO-24h group remained unchanged, which was basically the same as the content of H2O2 in the liver of normal aged mice. It indicates that oxidant treatment of aged mice during the active period ZT12-16 can restore the content of H2O2 in the liver of aged mice at ZT18. The specific experimental materials, experimental methods and experimental results are as follows:

[0094] (1) Experimental materials

[0095] Hydrogen peroxide content detection kit: Red Hydrogen Peroxide / Peroxidase AssayKit (A22188, ThermoFisher).

[0096] (2) Experimental methods

[0097] After two weeks of BSO drinking water, at ZT18, the mice were decapitated and sacrificed, 6 mice in each group. The mice were quickly dissected, and perfused with pre-cooled PBS buffer. The liver tissues of the mice were cut off, washed twice with pre-cooled PBS buffer, the residual liquid on the tissues was blotted dry with filter paper, placed in 1.5 mL EP tubes, frozen in liquid nitrogen for 30 s, and then stored in a -80 °C refrigerator for later use. The detection method of the content of H2O2 in the mouse liver was as described above.

[0098] (3) Experimental results

[0099] The experimental results are as Figure 5 shown. After two weeks of BSO drinking water, at ZT18, the content of H2O2 in the liver of aged mice in the Aged-BSO-4h group was significantly increased compared with that of normal aged mice, slightly lower than that of young mice; while the content of H2O2 in the liver of aged mice in the Aged-BSO-24h group remained unchanged, which was basically the same as the content of H2O2 in the liver of normal aged mice. It indicates that oxidant treatment of aged mice during the active period ZT12-16 can restore the content of H2O2 in the liver of aged mice at ZT18.

[0100] In a specific embodiment, the present invention detected the changes in the hepatic insulin pathway after time-dependent BSO administration in aged mice through Western blotting experiments, and found that compared with young mice, the expression levels of key proteins AKT and GSK3β in the insulin pathway in the liver of aged mice decreased, and the insulin-responsive proteins P-AKT and P-GSK3β decreased significantly, indicating that the insulin signaling pathway in aged mice was impaired. Compared with normal aged mice, the expression levels of AKT, P-AKT, and GSK3β in the livers of aged mice in the Aged-BSO-4h group were all increased to a certain extent, while the expression levels of P-AKT and P-GSK3β in the livers of aged mice in the Aged-BSO-24h group were basically the same as those of normal aged mice, indicating that time-dependent oxidant treatment could partially restore the activity of the hepatic insulin signaling pathway in aged mice. The specific experimental materials, experimental methods, and experimental results are as follows:

[0101] (1) Experimental materials

[0102] One-step PAGE gel rapid preparation kit (10%) (PG212, Yaenzyme); RIPA (P0013B, Beyotime); GAPDH antibody (ab8245, Abcam); AKT antibody (4691s, CST); P-AKT antibody (4060s, CST); GSK3β antibody (9315s, CST); P-GSK3β antibody (9323s, CST).

[0103] (2) Experimental methods

[0104] ① Liver sample treatment

[0105] Take 5 - 10 mg of liver samples and place them in a 1 mL EP tube containing 500 μL of RIPA cell lysate (1×cocktail, 1×PMSF), add grinding beads, and grind them in a grinder at 60 Hz for 180 sec at 4°C. Remove the grinding beads, use an ultrasonic crusher with a small probe, at 10% power, 15 sec On, 45 sec Off, for 5 min, and perform ultrasonic treatment in an ice-water bath until the sample is clarified. Then centrifuge at 4°C and 12,000 rpm for 10 min. Take the supernatant, add 5X SDS-PAGE loading buffer, boil in a boiling water bath for 10 min, and place it on ice for later use.

[0106] ② Gel preparation

[0107] Wash and dry the 1.5 cm glass plates, fix them in a vertical electrophoresis tank, prepare 10% separating gel and stacking gel according to the kit instructions, mix them by inverting up and down, minimize the generation of bubbles, and slowly pour them into the glass plates. The volume of each separating gel is 7 mL, and the stacking gel is 2 mL. Quickly insert the comb into the stacking gel and let it stand at room temperature for 30 min.

[0108] ③Electrophoresis

[0109] Pour a small amount of electrophoresis buffer into the electrophoresis tank first. Tilt the vertical electrophoresis instrument and place it in the electrophoresis tank to avoid air bubbles at the bottom. Then fill the electrophoresis instrument tank with electrophoresis buffer. Slowly pull out the comb teeth. Use a 1 mL syringe to adjust the walls of the gel wells to be vertical and suck out the residual gel in the wells. Connect the electrophoresis instrument. In a 4°C cold room, perform electrophoresis at a constant voltage of 60 V. When the Marker enters the separating gel and starts to separate, increase the voltage to 120 V and continue electrophoresis until the bromophenol blue reaches the bottom of the gel, then stop electrophoresis.

[0110] ④Transfer

[0111] Prepare the transfer solution according to the formula. Cut the PVDF membrane according to the size of the gel and immerse it in methanol to activate the PVDF membrane for later use. Place the transfer cassette in the transfer solution, open the transfer cassette, and lay the sponge, double-layer filter paper, PVDF membrane, gel, double-layer filter paper, and sponge in sequence, driving out the air bubbles between each layer. Clamp the transfer cassette and place it in the transfer rack with the gel layer at the positive electrode and the PVDF membrane layer at the negative electrode. In a 4°C cold room, perform ice-water bath electrotransfer at a constant current of 300 mA for 1 h.

[0112] ⑤Blocking

[0113] After the transfer is completed, place the PVDF membrane in a blocking solution of 5% skim milk and block it on a horizontal shaker at room temperature for 1 h.

[0114] ⑥Primary antibody hybridization

[0115] Rinse the blocked PVDF membrane 3 times with TBST buffer for 5 min each time to wash away the blocking solution on the membrane. Incubate it overnight at 4°C in a horizontal shaker with an appropriate primary antibody.

[0116] ⑦Secondary antibody hybridization

[0117] The next morning, take out the membrane incubated with the primary antibody and wash it 3 times with TBST buffer at room temperature for 5 min each time. Transfer the membrane to the corresponding secondary antibody (dilute the secondary antibody 1:5000 with a 5% skim milk blocking solution) and incubate it slowly on a horizontal shaker at room temperature for 2 h, then wash it 3 times with TBST buffer for 10 min each time.

[0118] ⑧Chemiluminescent imaging

[0119] Prepare the luminescent solution by mixing solution A and solution B in a volume ratio of 1:1, mix well, and place it in the dark. Drop the luminescent solution on the PVDF membrane and perform imaging with a chemiluminescence imager (Tanon), keeping it as dark as possible throughout the process.

[0120] (3) Experimental results

[0121] The experimental results are as Figure 6As shown, compared with young mice, the expression levels of key proteins AKT and GSK3β in the insulin pathway in the liver of old mice decreased, and the insulin-responsive proteins P-AKT and P-GSK3β decreased significantly, indicating that the insulin signaling pathway in old mice was damaged. In the old mice in the Aged-BSO-4h group, the expression levels of AKT, P-AKT, and GSK3β in the liver were all increased to a certain extent compared with those in normal old mice, while the expression levels of P-AKT and P-GSK3β in the liver of the old mice in the Aged-BSO-24h group were basically the same as those in normal old mice, indicating that time-dependent oxidant treatment could partially restore the activity of the insulin signaling pathway in the liver of old mice.

[0122] The present invention firstly discovers that treating with buthionine sulfoximine (BSO) during the period of reduced H2O2 in the liver (i.e., the active period ZT12-16) can restore the insulin sensitivity of old mice. By studying the changes in the redox system during the aging process from the time dimension and performing time-dependent oxidant treatment in response to its rhythm changes, the present invention realizes the restoration of the rhythm of the key signaling molecule H2O2 in the redox system of the liver of old mice and the treatment of insulin sensitivity injury. Based on this, the present invention has developed a brand-new intelligent diagnosis and treatment system, device, and computer-readable storage medium for old insulin sensitivity injury based on Internet medical services and precision, which can not only achieve precise intervention of buthionine sulfoximine, but also achieve the restoration of insulin sensitivity injury caused by aging and the treatment of type II diabetes in the elderly, and has broad clinical application prospects.

[0123] Figure 2 This is an intelligent diagnosis and treatment device for old insulin sensitivity injury based on Internet medical services provided by an embodiment of the present invention. The device includes a memory and a processor; the memory is used to store program instructions; the processor is used to call the program instructions, and when the program instructions are executed, it is used to execute the following methods:

[0124] S101: Obtain data on diagnostic elements of old insulin sensitivity injury related to the person to be diagnosed. The diagnostic elements of old insulin sensitivity injury include data on the expression levels of key proteins AKT and GSK3β in the insulin pathway in the liver, and data on the expression levels of insulin-responsive proteins P-AKT and P-GSK3β in the liver;

[0125] S102: Based on the data on diagnostic elements of old insulin sensitivity injury related to the person to be diagnosed, determine whether the person to be diagnosed is a patient with old insulin sensitivity injury;

[0126] If the expression levels of the key proteins AKT and GSK3β in the liver insulin pathway of the person to be diagnosed are significantly lower than those of young and healthy subjects, and the expression levels of the liver insulin-responsive proteins P-AKT and P-GSK3β are significantly lower than those of young and healthy subjects, then the person to be diagnosed is a patient with senile insulin sensitivity impairment;

[0127] S103: When the person to be diagnosed is a patient with senile insulin sensitivity impairment, select the following treatment plan: administer a therapeutically effective amount of buthionine sulfoximine to the person to be diagnosed during the period when the H2O2 in the liver of the person to be diagnosed decreases.

[0128] Furthermore, the period when H2O2 decreases is ZT12-16.

[0129] Furthermore, the concentration of buthionine sulfoximine is 20 Mm.

[0130] In some embodiments, the intelligent diagnosis and treatment device for senile insulin sensitivity impairment based on Internet medical services may further include: an input device and an output device.

[0131] In some embodiments, the memory, the processor, the input device and the output device may be connected by a bus or other means. Figure 3 The example shown is in the form of a bus connection; wherein, the memory is used to store program instructions; the processor is used to call the program instructions, and when the program instructions are executed, it is used to implement the above method.

[0132] In some embodiments, the memory can be understood as any device for storing programs, and the processor can be understood as a device for using programs.

[0133] The embodiment of the present invention also provides a computer-readable storage medium, on which a computer program is stored, and when the computer program is executed by a processor, the following method is implemented:

[0134] S101: Obtain data on diagnostic elements for senile insulin sensitivity impairment related to the person to be diagnosed, and the diagnostic elements for senile insulin sensitivity impairment include the expression level data of the key proteins AKT and GSK3β in the liver insulin pathway, and the expression level data of the liver insulin-responsive proteins P-AKT and P-GSK3β;

[0135] S102: Based on the data on diagnostic elements for senile insulin sensitivity impairment related to the person to be diagnosed, determine whether the person to be diagnosed is a patient with senile insulin sensitivity impairment;

[0136] If the expression levels of the key proteins AKT and GSK3β in the liver insulin pathway of the subject to be diagnosed are significantly lower than those of young and healthy subjects, and the expression levels of the liver insulin-responsive proteins P-AKT and P-GSK3β are significantly lower than those of young and healthy subjects, then the subject to be diagnosed is a patient with senile insulin sensitivity impairment;

[0137] S103: When the subject to be diagnosed is a patient with senile insulin sensitivity impairment, select the following treatment plan: administer a therapeutically effective amount of buthionine-sulfoximine to the subject during the period when the liver H2O2 of the subject decreases.

[0138] When the subject to be diagnosed is not a patient with senile insulin sensitivity impairment, there is no need to select the following treatment plan: administer a therapeutically effective amount of buthionine-sulfoximine to the subject during the period when the liver H2O2 of the subject decreases.

[0139] The embodiments of the present invention also provide a method for treating senile insulin sensitivity impairment or type II diabetes or a time-dependent therapy for senile insulin sensitivity impairment. The method includes administering a therapeutically effective amount of buthionine-sulfoximine to a subject in need thereof during the period when the liver H2O2 of the subject decreases.

[0140] In some embodiments, the subject refers to any animal, and also refers to humans and non-human animals. Non-human animals include all vertebrates, for example, mammals such as non-human primates (especially higher primates), sheep, dogs, rodents (such as mice or rats), guinea pigs, goats, pigs, cats, rabbits, cows, and any domestic animal or pet; and non-mammals such as chickens, amphibians, reptiles, etc. In a preferred embodiment, the subject is a human.

[0141] Those skilled in the art can clearly understand that for the convenience and conciseness of description, the specific working processes of the systems, devices, and units described above can refer to the corresponding processes in the foregoing method embodiments and will not be elaborated herein.

[0142] In the several embodiments provided in the present application, it should be understood that the disclosed systems, devices, and methods can be implemented in other ways. For example, the device embodiments described above are merely illustrative. For example, the division of the units is only a logical function division, and there can be other division methods in actual implementation. For example, multiple units or components can be combined or integrated into another system, or some features can be ignored or not executed. Another point, the couplings, direct couplings, or communication connections shown or discussed with each other can be indirect couplings or communication connections through some interfaces, devices, or units, and can be in electrical, mechanical, or other forms.

[0143] The unit described as a separation component may or may not be physically separated. The component displayed as a unit may or may not be a physical unit, that is, it may be located in one place or distributed across multiple network units. Some or all of the units can be selected according to actual needs to achieve the purpose of the solution of this embodiment.

[0144] In addition, in each embodiment of the present invention, each functional unit can be integrated into a processing unit, can exist separately as individual physical units, or two or more units can be integrated into one unit. The above integrated units can be implemented in the form of hardware or in the form of software functional units.

[0145] Those of ordinary skill in the art can understand that all or part of the steps in the various methods of the above embodiments can be completed by instructing relevant hardware through a program, and the program can be stored in a computer-readable storage medium. The storage medium can include: read-only memory (ROM, Read Only Memory), random access memory (RAM, Random Access Memory), magnetic disk or optical disc, etc.

[0146] The above has introduced in detail a computer device provided by the present invention. For those of ordinary skill in the art, according to the idea of the embodiments of the present invention, there will be changes in the specific implementation manner and application scope. In summary, the content of this specification should not be construed as a limitation to the present invention.

Claims

1. An intelligent diagnosis and treatment system for insulin sensitivity impairment in the elderly based on Internet medical services, characterized in that, The system includes: A diagnostic element information collector, configured to collect diagnostic element information related to a person to be diagnosed for senile insulin sensitivity injury and send the collected data results to a disease diagnosis matcher; A disease diagnosis matcher, configured to match the diagnostic element information of the diagnostic element information collector with disease types and send the matched data results to a treatment plan matcher; A treatment plan matcher, configured to determine the result matched by the disease diagnosis matcher and match and call a treatment plan according to the result, and then send the treatment plan data results to a disease diagnosis and treatment information outputter; A disease diagnosis and treatment information outputter, configured to output the data results received from the disease diagnosis matcher and the treatment plan matcher to a receiving unit; The diagnostic elements for senile insulin sensitivity injury include the expression level data of key proteins AKT and GSK3β in the hepatic insulin pathway, and the expression level data of hepatic insulin-responsive proteins P-AKT and P-GSK3β; In the disease diagnosis matcher, if the expression levels of key proteins AKT and GSK3β in the hepatic insulin pathway of the person to be diagnosed are significantly lower than those of young and healthy subjects, and the expression levels of hepatic insulin-responsive proteins P-AKT and P-GSK3β are significantly lower than those of young and healthy subjects, then the person to be diagnosed is matched as a patient with senile insulin sensitivity injury; In the disease diagnosis matcher, if the expression levels of key proteins AKT and GSK3β in the hepatic insulin pathway of the person to be diagnosed show no significant difference from those of young and healthy subjects, and the expression levels of hepatic insulin-responsive proteins P-AKT and P-GSK3β show no significant difference from those of young and healthy subjects, then the person to be diagnosed is matched as a non-senile insulin sensitivity injury person; In the treatment plan matcher, if the disease diagnosis matcher matches the person to be diagnosed as a patient with senile insulin sensitivity injury, then the following treatment plan is matched and called: administering a therapeutically effective amount of buthionine sulfoximine to the person to be diagnosed during the period when H2O2 in the liver of the person to be diagnosed decreases; In the treatment plan matcher, if the disease diagnosis matcher matches the person to be diagnosed as a non-senile insulin sensitivity injury person, then there is no need to match and call the following treatment plan: administering a therapeutically effective amount of buthionine sulfoximine to the person to be diagnosed during the period when H2O2 in the liver of the person to be diagnosed decreases.

2. The intelligent diagnosis and treatment system for insulin sensitivity impairment in the elderly based on Internet medical services according to claim 1, characterized in that, The period when H2O2 decreases is ZT12-16; The ZT12-16 is a time period from 12 hours to 16 hours after the start of light illumination.

3. The intelligent diagnosis and treatment system for insulin sensitivity impairment in the elderly based on Internet medical services according to claim 1, characterized in that, The concentration of buthionine sulfoximine is 20 mmol / L.

4. The intelligent diagnosis and treatment system for elderly insulin sensitivity injury based on Internet medical services according to claim 1, characterized in that, The treatment plan can restore the insulin sensitivity of patients with senile insulin sensitivity injury.

5. The intelligent diagnosis and treatment system for elderly insulin sensitivity injury based on Internet medical services according to claim 1, characterized in that, The receiving unit is a display screen, a computer client, a mobile phone client or a tablet computer.

6. An intelligent diagnosis and treatment device for insulin sensitivity impairment in the elderly based on Internet medical services, characterized in that, The device includes a memory and a processor; the memory is used for storing program instructions; the processor is used for calling the program instructions, and when the program instructions are executed, it is used to execute the following method: S101: Obtain the data of diagnostic elements for senile insulin sensitivity injury related to the person to be diagnosed. The diagnostic elements for senile insulin sensitivity injury include the expression data of key proteins AKT and GSK3β in the hepatic insulin pathway, and the expression data of hepatic insulin-responsive proteins P-AKT and P-GSK3β. S102: Based on the data of diagnostic elements for senile insulin sensitivity injury related to the person to be diagnosed, determine whether the person to be diagnosed is a patient with senile insulin sensitivity injury. If the expression levels of key proteins AKT and GSK3β in the hepatic insulin pathway of the person to be diagnosed are significantly lower than those of young healthy subjects, and the expression levels of hepatic insulin-responsive proteins P-AKT and P-GSK3β are significantly lower than those of young healthy subjects, then the person to be diagnosed is a patient with senile insulin sensitivity injury. S103: When the person to be diagnosed is a patient with senile insulin sensitivity injury, select the following treatment plan: Administer a therapeutically effective amount of buthionine sulfoximine to the person to be diagnosed during the period when H2O2 in the liver of the person to be diagnosed decreases.

7. The intelligent diagnosis and treatment device for insulin sensitivity impairment in the elderly based on Internet medical services according to claim 6, characterized in that, The period when H2O2 decreases is ZT12-16. The ZT12-16 is the time period from 12 hours to 16 hours after the start of light illumination.

8. The intelligent diagnosis and treatment device for insulin sensitivity impairment in the elderly based on Internet medical services according to claim 6, characterized in that, The concentration of buthionine sulfoximine is 20 mmol / L.

9. A computer-readable storage medium having a computer program stored thereon, characterized in that, When the computer program is executed by a processor, it implements the following method: S101: Obtain the data of diagnostic elements for senile insulin sensitivity injury related to the person to be diagnosed. The diagnostic elements for senile insulin sensitivity injury include the expression data of key proteins AKT and GSK3β in the hepatic insulin pathway, and the expression data of hepatic insulin-responsive proteins P-AKT and P-GSK3β. S102: Based on the data of diagnostic elements for senile insulin sensitivity injury related to the person to be diagnosed, determine whether the person to be diagnosed is a patient with senile insulin sensitivity injury. If the expression levels of key proteins AKT and GSK3β in the hepatic insulin pathway of the person to be diagnosed are significantly lower than those of young healthy subjects, and the expression levels of hepatic insulin-responsive proteins P-AKT and P-GSK3β are significantly lower than those of young healthy subjects, then the person to be diagnosed is a patient with senile insulin sensitivity injury. S103: When the person to be diagnosed is a patient with senile insulin sensitivity injury, select the following treatment plan: Administer a therapeutically effective amount of buthionine sulfoximine to the person to be diagnosed during the period when H2O2 in the liver of the person to be diagnosed decreases.

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