Patent Application
20240024287
This application is based upon and claims priority to Chinese Patent Application No: 202210866348.3, filed on Jul. 22, 2022, the entire contents of which are incorporated herein by reference.
The present invention relates to the technical field of insulin resistance, and in particular to an application of Anle138b in a drug for improving diet-induced insulin resistance.
As a core concept of metabolic syndromes, insulin resistance accompanies with the occurrence and development of the entire course of metabolic diseases, such as diabetes and obesity, as well as the occurrence and development of cardiovascular and other complications. In recent years, studies have found that in addition to peripheral insulin resistance, the pathogenic role of central insulin resistance has gradually become more important. Since insulin, insulin receptors and their signaling pathways are widely distributed in brains, especially dominated by the distribution in the hypothalamus for energy metabolism regulation, in the central nervous system, in addition to regulating the expression of neuropeptides and participating in the production of hepatic glucose, insulin, insulin receptors and their signaling pathways also play important roles in the normal metabolism of neurons and the function of tactile transmission, and it has been found that insulin resistance accompanies with a variety of neuronal dysfunction diseases. Several epidemiological studies have shown that insulin resistance increases the risk of dementia and AD. Nutrition-induced insulin resistance significantly affects neural insulin signaling and leads to cognitive impairment. Because insulin resistance is closely related to functions of the nervous system, more scholars now refer to AD as type 3 diabetes mellitus.
Anle138b, as a highly active low molecular weight compound of 3,5-diphenyl-pyrazole (DPP) derivatives, has high blood-brain barrier permeability, has significant curative effects on regulating intracranial amyloid deposition in both prevention and treatment, can directly interfere with the pathological aggregation of amyloid proteins such as prion, α-synuclein and Tau protein, and can improve the pathological progression of animal models of neurodegenerative diseases, such as multiple system atrophy, Parkinson's disease, and Alzheimer's disease.
The present invention is intended to provide an application of Anle138b in a drug for improving diet-induced insulin resistance. Anle138b has obvious ameliorative effects on both preventing and treating insulin resistance, and can reduce the intake efficiency and the body weight. Besides, Anle138b can effectively improve diet-induced ectopic fat storage and adipose tissue hypertrophy, and the effect of Anle138b on improving the insulin resistance is independent of the reduction of food intake.
In order to achieve the above objective, the present invention provides an application of Anle138b in a drug for improving diet-induced insulin resistance.
Preferably, improvement of insulin resistance includes prevention or treatment for improvement of insulin resistance.
An application of Anle138b in preparing a drug for reducing the food intake and body weight of patients with insulin resistance diseases.
Preferably, the insulin resistance is ameliorated by reducing the high-fat feeding efficiency and restoring insulin reactivity.
An application of Anle138b in preparing a drug for alleviating hepatic and pancreatic steatosis induced by insulin resistance.
Preferably, alleviating hepatic and pancreatic steatosis induced by insulin resistance is unrelated to reducing the food intake.
An application of Anle138b in preparing a drug for reducing the blood lipid of patients with insulin resistance diseases.
An application of Anle138b in preparing a drug for suppressing appetite in the process of losing weight.
Therefore, the application of Anle138b in a drug for improving diet-induced insulin resistance is adopted in the present invention, and it is found by judging the therapeutic effect of Anle138b on the high-fructose-induced rat insulin resistance models that the intraperitoneal injection of Anle138b 1 mg/kg for 2 weeks can improve the hyperinsulinemia of the rats and reverse the high-fructose-induced insulin resistance of the rats.
The technical solutions of the present invention will be further described in detail below in combination with the drawings and the embodiments.
The technical solutions of the present invention will be further described below in combination with the drawings and the embodiments.
Unless defined otherwise, all terms (including technical or scientific terms) used in the present disclosure have the same meaning as commonly understood by those of ordinary skill in the art to which the present disclosure belongs. It should also be understood that the terms defined in general-purpose dictionaries should be construed as having meanings consistent with their meanings in the context of the related art, and should not be interpreted in an idealized or highly formalized sense, unless explicitly defined in such a way in this context.
For those skilled in the art, it is obvious that the present invention is not limited to the details of the above exemplary embodiment, and the present invention can be implemented in other specific forms under the situation of not deviating from the main idea or basic features of the present invention. Therefore, from any point of view, the embodiment shall be regarded as exemplary and non-limiting. The scope of the present invention is defined by the attached claims rather than the above description.
8-week-old male C57BL/6J rats were selected as experimental animals. The rats were adaptively fed for 1 week after purchase, and then weighed for the fasting body weight. The healthy rats weighing 22-24 g were selected and randomly divided into groups according to the experimental flow design. (1) The rats were fed with normal feed and high-fructose feed for 8 weeks; (2) the rats were fed with high-fat feed for 12 weeks; (3) the rats were fed with high-fat feed for 8 weeks as a preventive experiment. Fasting body weights were measured and recorded regularly every week, and the rats were guaranteed to have free access to water and food during feeding. The food intake, water intake, and free movement of the model rats were continuously observed, and the rats with stunted growth, rough hair, and rickety posture after eating high-fructose and high-fat diets were screened out. In the experiment later, the rats were fed with normal feed or high-fructose/high-fat feed during the 2 weeks of intraperitoneal injection to ensure the consistency before and after administration.
Formula of high-fructose feed: the high-fructose feed had a fructose calorie of 66.7%, a carbohydrate calorie of 66.9%, a fat calorie of 12.9% and a protein calorie of 20.2%, with a total calorie of 3.6 kcal/g, and the high-fructose feed was provided by Jiangsu Medison Biomedical Co., Ltd.
High-fat formula: the high-fat purified feed had a fat energy supply ratio of 45%, wherein the energy ratio of protein was 20% and the energy ratio of carbohydrate was 35%. A total energy-to-mass ratio was 4.73 kcal/g. The feed was provided by Beijing Huafukang Biotechnology Co., Ltd.
Anle138b solutions: The drug doses required for 1 week of intervention for each group of rats were calculated, Anle138b drug concentrations of 1 mg/kg (0.01%), 3 mg/kg (0.03%) and 5 mg/kg (0.05%) were prepared, Anle138b powder was weighed and then added in 10% DMSO, 40% PEG300, 5% Tween-80 and 45% saline in sequence, each mixture was evenly blended by vortex into a clear solution, and each Anle138b solution was separately bottled according to the daily dose and stored in a refrigerator at 4° C.
L-THP solution: The drug dose required for 1 week of intervention for each group of rats was calculated, L-THP drug concentrations of 3 mg/kg (0.03%) was prepared, light yellow L-THP powder was weighed and then added in 10% DMSO, 40% PEG300, 5% Tween-80 and 45% saline in sequence, each mixture was evenly blended by vortex into a clear solution, and each Anle138b solution was separately bottled according to the daily dosage and stored in a refrigerator at 4° C.
The 8-week-old adult rats were modeled by changing the diets after 1 week of adaptive feeding. Blood was collected from the right orbital venous plexus with a glass tube at 8 weeks and 12 weeks after model establishment and after fasting (>12 h), and blood glucose levels were measured with a blood glucose tester (Sannuo GA-3); and the insulin resistance mode was judged by the subsequent determination of metabolic indexes, such as glucose tolerance test by intraperitoneal injection, fasting serum insulin (enzyme-linked immunosorbent assay, Raybiotech), serum triglyceride (GPO-PAP single-reagent microplate method, Nanjing Jiancheng) and cholesterol (GPO-PAP single-reagent microplate method, Nanjing Jiancheng).
(1) The normal rats were randomly divided into two groups: blank control group (n=6) and drug control group (n=7); the rats in the normal control group were intraperitoneally injected with a similar volume of solvent and Anle138b (3 mg/kg, 0.03%) according to the body weight for 2 weeks.
The rats were fed with 60% high-fructose diet for 8 weeks to establish insulin resistance models, and the rats fed with normal diet provided by the animal room of Shantou University Medical College were used as controls. After fasting, orbital blood was collected, and fasting blood glucose, serum insulin, glucose tolerance were determined to judge whether the modeling was successful. According to an insulin resistance index (HOMA-IR) as the core judgment index, the successfully modeled rats were enrolled by screening the IR index of >3 and randomly divided into groups to explore the influence of different concentrations of Anle138b on the rat insulin resistance models. The concentration gradients for different groups were 0 mg/kg (injection solvent, namely the model control group), 1 mg/kg, 3 mg/kg and 5 mg/kg clear Anle138b solution, as well as the L-THP 3 mg/kg positive drug control group. The rats were administrated by intraperitoneal injection for 2 weeks continuously, once a day.
Anle138b 1 mg/kg, 3 mg/kg and 5 mg/kg could all effectively reduce the fasting blood glucose, improve hyperinsulinemia, and reduce the insulin resistance value. In addition to reducing the epididymal fat weight index, Anle138b 1 mg/kg, 3 mg/kg and 5 mg/kg pathologically could also effectively improve the liver steatosis and epididymal adipose tissue hyperplasia caused by high-fructose diet, and could alleviate the inflammatory response of tissues. This indicated that Anle138b had a therapeutic effect on the high-fructose-induced insulin resistance of rats.
The body weights of the IR model group and the normal control group during modeling are as shown in Table 1, and the development trend is as shown in
In the self-control experiment of the normal control group before and after administration, there was no statistical significance for the difference in the body weight of the control group before the solvent was administered and after 2 weeks from the intraperitoneal injection of the solvent. The body weight after administration of the Anle138b 3 mg/kg drug decreased as compared with the body weight before administration, and there was statistical significance for the difference. This indicated that the Anle138b drug had an effect on reducing the body weight of normal rats.
There was no statistical significance for the difference between the body weights before and after administration of the solvent in the high-fructose model group. The efficacy of the drug on the high-fructose model group was determined after excluding the influence of the intraperitoneal injection of the solvent on the body weight of the rats.
As shown in
The influence of different concentrations of Anle138b on the metabolic indexes of the high-fructose-induced rat insulin resistance models is as shown in Table 3. High fructose treatment for 2 weeks could increase the fasting blood glucose levels of the rats (as shown in
For the rats treated with high fructose for 2 weeks, the fasting blood glucose levels of the Anle138b drug treatment group after intervention at 1 mg/kg, 3 mg/kg and 5 mg/kg were all lower than those of the model solvent control group. Therefore, it could be known that the Anle138b drug could effectively reduce the elevated blood glucose in the diet-induced model of insulin resistance. The fasting blood glucose level of the drug control group (normal rats) treated with Anle138b 3 mg/kg for 2 weeks was not significantly different from that of the blank control group, so Anle138b could only reduce the blood glucose of the rat insulin resistance models but had no obvious influence on the blood glucose level of the normal rats. The blood glucose of the positive control group after intervention with L-THP 3 mg/kg was significantly lower than that of the solvent control group, and there was statistical significance for the differences.
The changes in the serum blood lipids were measured after the models were fed for 8 weeks. The triglyceride value of the normal control group was (0.6733±0.6733) mmol/L (n=7); the triglyceride value of the high-fructose model group was (0.6827±0.2120) mmol/L (n=7); there was no statistical significance for the difference between the two groups (not shown in the figure).Considering that the high fructose feeding for 8 weeks failed to reach the level of increasing serum triglyceride, the elevated serum triglyceride could prolong the feeding time or serum blood lipid indicators which were more sensitive at the early period of high fructose feeding were measured, such as serum free fatty acids, latter liver triglyceride, etc., so changes in the blood lipids were not compared currently, and then feeding evaluation could be performed by changing high-fat feed to improve the time efficiency of feeding.
The fasting insulin value of the normal control group was (10.28±3.72) (n=8), the fasting insulin value after intervention with Anle138b 3 mg/kg for 2 weeks was (15.08±13.14) μIU/mL (n=6), and both were within a normal range. The fasting insulin value of the high-fructose model group was significantly higher than that of the normal group, and there was statistical significance for the difference (P<0.0001), indicating that feeding with 60% high-fructose diet for 8 weeks could lead to hyperinsulinemia of rats. Combined with the changes in the blood glucose, it was considered that the insulin resistance models were successfully established.
The insulin resistance value was calculated in combination with the method for calculating the insulin resistance index (HOMA-IR) in the homeostasis model assessment method. The insulin resistance value of the normal control group was (2.055±0.58) μIU/mL*mmol/L, and that of the normal group after Anle138b intervention was (1.1689±1.153) μIU/mL*mmol/L, both were within a normal range, and there was no statistical significance for the difference. At the same time, the resistance value of the high-fructose model group was significantly higher than that of the normal group, and there was statistical significance for the difference (P<0.0001).
Comparing the insulin resistance values after intervention with different concentrations of Anle138b, the insulin resistance value after intervention with Anle138b 1 mg/kg was significantly lower than that of the model control group; the insulin resistance value after intervention with Anle138b 3 mg/kg was significantly lower than that of the model control group; the insulin resistance value after intervention with Anle138b 5 mg/kg was significantly lower than that of the model control group. The insulin resistance value of the positive control group after intervention with L-THP 3 mg/kg was lower than that of the model control group.
According to the comprehensive judgment in combination with the blood glucose, the insulin value and the insulin resistance value, Anle138b could effectively improve the high-fructose-induced insulin resistance of rats in a dose-dependent manner. More indexes would be further judged for corroboration subsequently.
The results of the glucose tolerance test are as shown in
The right epididymal fat weight index was calculated by the right epididymal fat weight/fasting body weight, and it was obtained that the fat weight index of the normal solvent control group was 0.005624±0.0007023 (n=8); the fat weight index of the high-fructose solvent group was 0.01082±0.001067 (n=7); compared with the normal solvent group, the fat weight per unit body weight was significantly increased, and there was statistical significance for the difference (P<0.001).
The fat weight index after intervention with Anle138b 1 mg/kg was 0.008918±0.0003788 (n=7), which was lower than that of the model solvent control group, P<0.01; the fat weight index after intervention with Anle138b 3 mg/kg was 0.007657±0.002366 (n=6), P<0.01; the fat weight index after intervention with Anle138b 5 mg/kg was 0.007971±0.001420 (n=5), P<0.01; the fat value was close with that of the L-THP positive control group (0.007510±0.002279, n=6), and as shown in
After 8 weeks of high-fructose feeding, the livers of the rats were pathologically examined; under a low-power microscope, the unclear demarcation of hepatic lobules of the rats could be observed; under a high-power microscope, the structural damage of the central hepatic vein, the disordered arrangement of hepatic cords, and the microcystic changes of fat accumulation in hepatocytes, namely hepatic steatosis, could be observed, the fusion of small lipid droplets and the death of hepatocytes could be partially observed; after intervention with Anle138b, it could be observed that the hepatocyte damage was alleviated in a dose-dependent manner, the hepatic steatosis was significantly alleviated, the central vein structure and the arrangement structure of the hepatic cord were restored, and the infiltration of inflammatory cells was reduced; this indicated that the drug application could effectively alleviate the hepatocyte injury induced by high fructose, as shown in
The pathological examination of the epididymal adipose tissue showed that in the high-fructose group the fat structure was disordered, the coronal structure formed by aggregated inflammatory cells was observed in the fat space, the fat cells were enlarged in different sizes and irregular shapes, and part of the cells were fused into big lipid droplets; after 2 weeks of drug treatment with Anle138b, it could be seen that with the increase of the drug concentration, the adipocytes were reduced in sizes and arranged regularly, and the tubular structure was reduced; the pathological examination of the adipose tissue showed that the drug reduced the nutritional burden of adipocytes by reducing the food intake to improve the early diet-induced fat inflammation and slow down the disease progression.
The pathological examination of pancreas after 8 weeks of high-fructose feeding showed no obvious structural damage of islets and acini.
The rat IR models were randomly divided into two groups, the rats were administrated with Anle138b 3 mg/kg (n=6, 0.03%) and 5 mg/kg (n=6, 0.05%) for 2 weeks continuously, once a day.
Anle138b 3 mg/kg and 5 mg/kg could effectively reduce the food intake and body weight, reduce the feeding efficiency, reduce the fasting blood glucose and triglyceride, improve hyperinsulinemia, and reduce the insulin resistance value. Anle138b 3 mg/kg and 5 mg/kg could lower the liver weight index and the epididymal fat weight index. Pathologically, Anle138b 3 mg/kg and 5 mg/kg could significantly improve liver steatosis, pancreatic acinar steatosis, pathological hyperplasia of epididymal adipose tissue and intermuscular adipose hyperplasia caused by high fat, and reduce the inflammatory response of various tissues. This experiment showed that Anle138b had a therapeutic effect on the high-fat-induced insulin resistance of rats.
After the models were determined to be successful, the average value of food intake of each rat in the first two days was taken as its own food intake base. After the drug intervention, the daily food intake and body weight were measured and compared with their own bases for comparison of the standardized food intake and body weight of each rat.
The average daily food intake of the high-fat fed rats was (2.789±0.3928) g/day (n=5); the average daily food intake of the Anle138b 3 mg/kg group was (1.946±0.4777) g/day (n=5), which was significantly lower than that of the model group, and there was statistical significance for the difference (P<0.001); the average daily food intake of the Anle138b 5 mg/kg group was significantly lower than that of the model group, and there was statistical significance for the difference (P<0.001). As shown in
For the high-fat fed rats, the weight before intervention was (31.01±1.332) g; the weight after intervention with Anle138b 3 mg/kg was (28.97±1.657) g (n=6), and the weight after intervention with Anle138b 5 mg/kg was (28.9±1.685) g (n=5); compared with the model group, the body weight was decreased, and there was statistical significance for the differences (P<0.05).The body weight decreased after the drug intervention, and the decrease was more obvious at high doses.
The feeding efficiency of each rat was obtained by the daily body weight change/energy consumption (g/calories/rat/day); the feeding efficiency of the high-fat feeding model group was 0.02491±0.008717; the feeding efficiency was −0.007788±0.01949 after intervention with Anle138b 3 mg/kg and −0.01528±0.01287 after intervention with Anle138b 5 mg/kg; compared with the model group, it could be seen that Anle138b could increase the feed utilization, reduce the consumption of high-fat feed, and reduce the feeding efficiency.
As shown in Table 4 and
The glucose tolerance test was performed, and the results are as shown in
The liver weight index of the model group was 0.05058±0.004 (n=7); the liver weight index decreased to 0.04475±0.003 after intervention with Anle138b 3 mg/kg; the liver weight index decreased to 0.04344±0.0026 after intervention with Anle138b 5 mg/kg; there was statistical significance for the differences. The fat weight index of the model group was 0.02148±0.00702 (n=6); the fat weight index decreased to 0.01317±0.004212 after intervention with Anle138b 3 mg/kg; the fat weight index decreased to after intervention with Anle138b 5 mg/kg. This indicated that the drug could effectively relieve and reduce the weight of liver and fat after application, as shown in
The results of the pathological examination showed that after 12 weeks of high-fat feeding, unclear demarcation and structural disorder of hepatic lobules of the rats could be observed, there was obvious fat infiltration merging into circular lipid droplets of different sizes in the liver, the central hepatic vein has abnormal structure, accompanied by the infiltration of inflammatory cells and the disordered arrangement of hepatic cord; after intervention with Anle138b 3 mg/kg and 5 mg/kg, respectively, the hepatic steatosis was significantly alleviated, no large lipid droplets were seen, and the central vein structure and hepatic cord arrangement structure recovered, and the infiltration of inflammatory cells was reduced; this indicated that the drug application could effectively alleviate the hepatocyte injury induced by high-fat diet.
Different from the pathological examination of pancreas after 8 weeks of high-fructose feeding, disordered arrangement of pancreas islet cells, enlarged capillaries in the islets, damage to the normal structure of pancreatic acinar cells, locally digested acinar cells and obvious acinar steatosis could be observed after 12 weeks of high-fat feeding; after intervention with Anle138b 3 mg/kg and 5 mg/kg, it could be seen that as the dose increases, the acinar steatosis was significantly alleviated, the structure was restored as compared with that in the model group, and the capillaries in the islet cells were reduced; this indicated that the drug application could effectively alleviate the pancreatic cell damage induced by high-fat diet.
The pathological examination of the epididymal adipose tissue showed that in the model group, the fat structure was disordered, the coronal structure formed by aggregated inflammatory cells was observed in the fat space and obviously increased as compared with that in the high-fructose feeding group, a part formed into a cord shape along the cells, the fat cells were enlarged in different sizes, the normal structure was destroyed; after 2 weeks of drug treatment with Anle138b, it could be seen that with the increase of the drug concentration, the adipocytes were reduced in sizes and arranged regularly, and the tubular structure was reduced.
During the 12 weeks of high-fat diet feeding, intramuscular fat infiltration of skeletal muscle cells was observed, intermuscular fat manifestations occurred, and meanwhile, the disordered arrangement of muscle cords and the infiltration of inflammatory cells also occurred; after the drug intervention for 2 weeks, the intermuscular steatosis was relieved, but the arrangement structure of muscle cords did not improve significantly, and there were also manifestations of inflammatory infiltration, as shown in
Rats fed with high-fat diet for 5 weeks were randomly divided into high-fat feeding control group (n=6) and drug intervention group with Anle138b 5 mg/kg, and the rats were administered for 3 weeks continuously, once every two days.
The preventive administration with Anle138b 5 mg/kg could reduce the food intake of rats, inhibit the body weight gain, effectively inhibit the increase of fasting blood glucose, insulin, triglyceride and cholesterol, improve the insulin resistance, and improve the glucose tolerance response. In pathological detection, Anle138b could effectively alleviate the tissue structure damage and inflammatory response to pancreas and visceral fat induced by high-fat diet. This indicated that Anle138b had preventive effect on insulin resistance induced by high-fat diet in the rat models.
In the prevention experiment, the changes in the food intake and body weight of rats were monitored. As shown in
As shown in Table 5, the fasting blood glucose of the model group was (9.283±0.66) mmol/L (n=6), the fasting blood glucose of the drug group was (6.313±0.63) mmol/L (n=8), which was significantly lower than that of the model group, and there was statistical significance for the difference (P<0.001); the triglyceride and the total cholesterol in the model group were (0.7024±0.21) mmol/L and (4.099±0.32) mmol/L, respectively; the triglyceride and the total cholesterol in the drug group decreased to (0.4687±0.05) mmol/L and (3.383±0.21) mmol/L; the fasting serum insulin of the model group was (47.67±12.61) μIU/mL, and the fasting serum insulin drug group decreased to (22.09±6.49) μIU/mL; the insulin resistance index was calculated, the insulin resistance index was (17.43±7.00) μIU/mL*mmol/L in the model group and (6.880±2.73) μIU/mL*mmol/L in the drug group, as shown in
The results of the glucose tolerance test are shown in
As shown in
The pathological examination showed that as shown in
The rats fed with high-fat diet for 8 weeks were divided into paired-feed group (n=7) (the food intake was the same as that of high-fat Anle138b 5 mg/kg/d) and control group continuously fed with high-fat diet.
There was no significant difference in the changes in the body weight measurement between the pair-feed group and the Anle138b 5 mg/kg group, indicating that the drug intervention induced body weight loss was caused by the decrease in the food intake. There was no decrease in the serum fasting blood glucose, triglyceride and insulin concentrations in the paired-feed group, indicating that simply reducing the food intake could not inhibit the glucose metabolism disorder and hyperinsulinemia caused by high-fat diet, also indicating that the effect of Anle138b on improving the insulin resistance was independent of suppressing the dietary decline. The comparison of the pathological examination showed that the paired-feed group could neither reduce the ectopic fat storage in liver and muscle, nor improve the vascular proliferation, structural disorder and acinar steatosis in pancreatic islets. In summary, Anle138b could alleviate the hepatic and pancreatic steatosis in rat insulin resistance models, and this function was independent of simply reducing the food intake; only its weight-reducing effect was associated with the reduction in the food intake.
Pair-feed (PF) group: the pair-feed group was set according to the daily food intake in the high-fat group after intervention with Anle138b 5 mg/kg, the rats were given the same amount of high-fat diet every day, and the changes in the body weight and the metabolic indexes were measured.
As shown in
The influence of Anle138b treatment+PF group on the metabolic indexes of rat insulin resistance models is as shown in
The comparison of the insulin resistance is as shown in
As shown in
As shown in
The pathological examination showed that, as shown in
Therefore, the application of Anle138b in a drug for improving diet-induced insulin resistance is adopted in the present invention, Anle138b has obvious improvement effects on both preventing and treating insulin resistance, and can reduce the intake efficiency and the body weight. Besides, Anle138b can effectively improve diet-induced ectopic fat storage and adipose tissue hypertrophy, and the effect of Anle138b on improving the insulin resistance is independent of the reduction of food intake.
Finally, it shall be stated that the embodiments above are only used for explaining, rather than restricting, the technical schemes of the present invention. Although the present invention is explained in detail referring to the preferred embodiments, those of skill in the art should be understood that, the technical schemes of the present invention can be modified or equivalently replaced, whereas the modifications or equivalent replacements will not make the modified technical scheme deviate from the spirit and scope of the technical schemes of the present invention.
| Number | Date | Country | Kind |
|---|---|---|---|
| 202210866348.3 | Jul 2022 | CN | national |
