Parameter positioning method and kit for inhibiting polycystic ovary syndrome (PCOS) complicated with epilepsy based on focused ultrasound

Abstract

This application relates to the field of focused ultrasound, and more particularly to a parameter localization method and kit for inhibiting PCOS-related epilepsy using focused ultrasound. The method includes: acquiring multiple preset power levels, each corresponding to different combinations of power and duty cycle; predicting the safe range of stimulation duration at different power levels based on prior experimental data and a biothermal model; setting several experimental parameter combinations with different preset doses based on the safe ranges; performing ultrasound stimulation on an experimental model based on the experimental parameter combinations and obtaining biological response results; fitting dose-response curves for each power level based on a preset dose-response relationship and the biological response results; determining the optimal dose range for each power level based on the dose-response curves, and evaluating the feasibility of the optimal dose range to determine the optimal parameter combination for focused ultrasound. This method efficiently determines a relatively accurate and effective range of ultrasound parameters while maintaining simple experimental operation, short cycle time, and low cost, providing support for personalized medicine.

Description

Technical Field

[0001] This application relates to the field of focused ultrasound, and in particular to a parameter localization method and kit for suppressing PCOS-related epilepsy based on focused ultrasound. Background Technology

[0002] Polycystic ovary syndrome (PCOS) is a common endocrine and metabolic disorder. Previous studies have shown a significantly higher prevalence of PCOS in women with epilepsy, approximately 15%–30%, which is 2–3 times higher than in the healthy population, suggesting a possible close comorbidity between the two conditions. Furthermore, clinical observations have found that women with both PCOS and epilepsy have a higher frequency of seizures than those with typical epilepsy (e.g., 22.2% vs 13.4%), suggesting that PCOS may promote susceptibility to epilepsy or increase seizure activity.

[0003] Building on this, animal experiments further support the aforementioned inverse correlation: letrozole-induced PCOS rats showed increased susceptibility to epilepsy, which decreased after anti-androgen treatment. Mechanistically, in PCOS, there is an increase in sympathetic nerve fibers surrounding small follicles in the ovary, and abnormal follicles tend to atresize, suggesting that the peripheral ovary and sympathetic axis may influence central excitability, thereby participating in the occurrence and development of epilepsy.

[0004] Focused ultrasound (FUS) is a non-invasive physical modulation technique that uses focused ultrasound to stimulate the ovaries through the body surface. The thermal and mechanical effects of focused ultrasound may promote the atresia of abnormal follicles and change the distribution of nerve endings, thereby achieving peripheral nerve modulation through afferent nerves and alleviating the susceptibility to epilepsy in PCOS patients.

[0005] However, the therapeutic effect of focused ultrasound is highly dependent on the precise combination of parameters such as power, duty cycle and frequency, simulated focal plane distance and stimulation duration. Different combinations of parameters may lead to drastically different biological effects, or even ineffective or adverse results. Currently, there is a lack of efficient and systematic methods to quickly explore effective parameter combinations suitable for suppressing PCOS-related epilepsy. Summary of the Invention

[0006] The main objective of this application is to provide a parameter localization method and kit for suppressing PCOS-related epilepsy based on focused ultrasound. To address the aforementioned technical problems, this application specifically adopts the following technical solution:

[0007] The first aspect of this application is to provide a parameter localization method for suppressing PCOS-related epilepsy based on focused ultrasound, wherein the parameters of the focused ultrasound include power, duty cycle, and stimulation duration, and the method includes:

[0008] S101: Acquire multiple preset power levels, each corresponding to a different combination of power and duty cycle; based on prior experimental data and biothermal models, predict the safe range of stimulation duration at different power levels.

[0009] S102, based on the safe range of stimulation duration for each level, several combinations of experimental parameters with different preset doses are set, wherein the dose is the product of power, duty cycle and stimulation duration;

[0010] S103, based on several combinations of the experimental parameters, the experimental model is subjected to ultrasonic stimulation to obtain the corresponding biological response results;

[0011] S104, based on a preset dose-effect relationship, according to the biological response results of each combination of experimental parameters at each level, the dose-effect curves at each level are fitted respectively; the preset dose-effect relationship includes: a. the dose-effect curves exhibit a single-peak distribution within the safe parameter range; b. the peak dose of the dose-effect curves is inversely correlated with the values ​​of power and duty cycle at each level;

[0012] S105, Based on the dose-effect curve, determine the preferred dose range for each level, and conduct a feasibility assessment of the preferred dose range for each level to determine the preferred parameter combination for focused ultrasound.

[0013] A second aspect of this application is to provide a test kit, the kit comprising:

[0014] The safe zone testing module is used to acquire multiple preset power levels, each corresponding to a different combination of power and duty cycle; based on prior experimental data and biothermal models, it predicts the safe zone of stimulation duration at different power levels.

[0015] The experimental parameter setting module is used to set several combinations of experimental parameters with different preset doses according to the safe range of stimulation duration for each level. The dose is the product of power, duty cycle and stimulation duration.

[0016] An ultrasound stimulation execution module is used to perform ultrasound stimulation on the experimental model based on several combinations of the experimental parameters, and obtain the corresponding biological response results.

[0017] The dose-response curve module is used to fit the dose-response curve for each level based on a preset dose-response relationship and the biological response results of each combination of experimental parameters at each level. The preset dose-response relationship includes: a. The dose-response curve exhibits a single-peak distribution within the safe parameter range; b. The peak dose of the dose-response curve is inversely correlated with the power and duty cycle values ​​at each level.

[0018] The preferred dose range module is used to determine the preferred dose range for each level based on the dose-effect curve, and to conduct a feasibility assessment of the preferred dose range for each level in order to determine the preferred parameter combination for focused ultrasound.

[0019] Beneficial effects:

[0020] This application provides a parameter localization method and kit for suppressing PCOS-related epilepsy based on focused ultrasound. By obtaining limited data through a simplified experiment and combining it with a preset dose-response relationship, the effective parameter range can be quickly locked. Under the premise of ensuring simple operation, short cycle and low cost, the relatively accurate and effective range of ultrasound parameters can be efficiently determined, providing support for personalized medicine and improving the feasibility and safety of treatment for PCOS-related epilepsy.

[0021] In the initial stage, by using a small amount of readily available prior experimental data (such as the known critical time for tissue damage at a certain setting) combined with a mature biothermal model, the safe range of stimulation duration at different settings can be quickly predicted, avoiding the high animal cost of traditional large-scale pre-experiments and greatly shortening the early exploration cycle.

[0022] In the experimental design phase, dosage was introduced as a unified metric, and a limited number of representative dosage groups (such as high, medium, and low doses) were set up. Each group contained parameter combinations with different levels but similar dosages (such as level 3 for 30 seconds, level 2 for 70 seconds, and level 1 for 100 seconds, all with a dosage of approximately 75). This allowed a small number of experiments to cover a wide parameter space, reducing the number of experimental operations and resource consumption. Furthermore, the experimental procedure was simple and efficient: ultrasound stimulation was performed every other day, alternating with seizure time testing. This eliminated the need for complex pathological sections, ex vivo organ manipulation, or invasive testing, allowing for rapid and intuitive data acquisition, further shortening the experimental cycle and reducing technical barriers and costs.

[0023] Based on this, by utilizing pre-defined dose-effect relationship assumptions (such as unimodal distribution, slope, peak value, etc.), the modeling uncertainty caused by limited experimental data is effectively compensated for, and dose-benefit curves for each dose level are quickly fitted based on limited experimental data. Although the dose-effect curves generated in this way may sacrifice some absolute accuracy, they can reliably reflect the overall trend, accurately define the potential optimal dose range, and achieve efficient transformation from minimally simplistic experiments to effective parameter derivation. Furthermore, if the preliminary fitting results have uncertainties (such as multimodal distribution or excessively wide range), fine exploration can be carried out by setting up high and low dose verification groups within the optimal range.

[0024] The parameter localization method for suppressing PCOS-related epilepsy based on focused ultrasound provided in this application is particularly suitable for personalized medical scenarios. For example, when combined with parameters such as frequency and simulated focal plane distance, which are determined based on the average ovarian depth and symptoms of a population, doctors or researchers can quickly explore and determine personalized effective parameters for specific populations (such as differences in ovarian depth due to different races and body types) or different PCOS symptoms. Attached Figure Description

[0025] To more clearly illustrate the technical solutions in the embodiments of this application or the prior art, the drawings used in the description of the embodiments or the prior art will be briefly introduced below. In all the drawings, similar elements or parts are generally identified by similar reference numerals. The elements or parts in the drawings are not necessarily drawn to scale. Obviously, the drawings described below are some embodiments of this application; for those skilled in the art, other drawings can be obtained based on these drawings without any creative effort.

[0026] Figure 1 This is a schematic flowchart of a parameter localization method for suppressing PCOS-related epilepsy based on focused ultrasound, provided in an embodiment of this application.

[0027] Figure 2 This application provides an embodiment of an epileptic seizure trend diagram during ultrasound stimulation;

[0028] Figure 3 This is a schematic diagram of dose-response curves at various levels provided in the embodiments of this application;

[0029] Figure 4 This is a schematic diagram of another dose-response curve at various levels provided in the embodiments of this application;

[0030] Figure 5 This is a schematic diagram of another dose-response curve at various levels provided in the embodiments of this application;

[0031] Figure 6 This is a graph showing the weight changes of the animal model provided in this application at key time points;

[0032] Figure 7 This is a comparison chart of serum hormone levels among the experimental groups provided in the embodiments of this application;

[0033] Figure 8 This is a schematic block diagram of a reagent kit provided in an embodiment of this application. Detailed Implementation

[0034] To make the objectives, technical solutions, and advantages of the embodiments of this application clearer, the technical solutions of the embodiments of this application will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only some, not all, of the embodiments of this application. All other embodiments obtained by those skilled in the art based on the embodiments of this application without creative effort are within the scope of protection of this application.

[0035] The flowchart shown in the attached diagram is for illustrative purposes only and does not necessarily include all content and operations / steps, nor does it necessarily have to be performed in the order described. For example, some operations / steps can be broken down, combined, or partially merged, so the actual execution order may change depending on the actual situation.

[0036] In this document, suffixes such as “module,” “part,” or “unit” used to denote elements are used only for illustrative purposes and have no specific meaning in themselves. Therefore, “module,” “part,” or “unit” may be used interchangeably.

[0037] In this document, the terms "upper," "lower," "inner," "outer," "front," "rear," "one end," and "the other end," etc., indicate the orientation or positional relationship based on the orientation or positional relationship shown in the accompanying drawings. They are used only for the convenience of describing this application and simplifying the description, and do not indicate or imply that the device or element referred to must have a specific orientation, or be constructed and operated in a specific orientation. Therefore, they should not be construed as limitations on this application. Furthermore, the terms "first" and "second" are used for descriptive purposes only and should not be construed as indicating or implying relative importance.

[0038] In this document, unless otherwise explicitly specified and limited, the terms "installed," "equipped with," and "connected," etc., should be interpreted broadly. For example, "connection" can be a fixed connection, a detachable connection, or an integral connection; it can be a mechanical connection, a direct connection, or an indirect connection through an intermediate medium; it can be a connection within two components. Those skilled in the art can understand the specific meaning of the above terms in this application based on the specific circumstances.

[0039] In this document, the term “and / or” includes any and all combinations of one or more of the listed related items.

[0040] In this article, the term "multiple" means two or more, that is, it includes two, three, four, five, etc.

[0041] It should be noted that, in this document, the terms "comprising," "including," or any other variations thereof are intended to cover non-exclusive inclusion, such that a process, method, article, or apparatus that comprises a list of elements includes not only those elements but also other elements not expressly listed, or elements inherent to such a process, method, article, or apparatus. Unless otherwise specified, an element defined by the phrase "comprising one..." does not exclude the presence of other identical elements in the process, method, article, or apparatus that includes that element.

[0042] As used in this specification, the term "approximately" typically means + / - the value. 5%, or more typically, the value + / 4%, more typically the value + / 3%, more typically the value + / 2%, or even more typically + / - of the stated value 1%, or even more typically + / - of the value 0.5%.

[0043] In this specification, certain embodiments may be disclosed in a range-bound format. It should be understood that this "range-bound" description is merely for convenience and brevity and should not be construed as a rigid limitation on the disclosed range. Therefore, the description of the range should be considered as having specifically disclosed all possible subranges and independent numerical values ​​within those ranges. For example, range 1... The description of 6 should be considered as having specifically disclosed subranges such as from 1 to 3, from 1 to 4, from 1 to 5, from 2 to 4, from 2 to 6, from 3 to 6, etc., as well as individual numbers within this range, such as 1, 2, 3, 4, 5, and 6. The above rules apply regardless of the breadth of the range.

[0044] As a non-invasive physical modulation method, the therapeutic effect of focused ultrasound highly depends on the precise coordination of several key parameters, including power, duty cycle, stimulation duration, frequency, and simulated focal plane distance.

[0045] Among these, power, also known as acoustic power, is the output intensity of the ultrasonic emission; duty cycle is the ratio of the ultrasonic pulse being on and off, such as a duty cycle of 10%, which means that 10 milliseconds are emitted every 100 milliseconds; stimulation duration is the duration of a single ultrasonic treatment (e.g., 30 seconds, 60 seconds, etc.) and / or the total duration of multiple ultrasonic treatments. Stimulation duration, power, and duty cycle together determine the total energy input (also known as dose).

[0046] Among these parameters, frequency refers to the acoustic frequency of ultrasound waves. Higher frequencies result in more precise focusing and reduced penetration depth; lower frequencies allow for deeper penetration but decrease focusing accuracy. The simulated focal plane distance is the preset depth position of the ultrasound focus within the body. Frequency and simulated focal plane distance need to be individually set based on individual anatomical structures (such as the depth of the ovary from the body surface) to ensure that energy is precisely focused on a specific area and to avoid accidental damage to surrounding tissues.

[0047] In this embodiment, power, duty cycle, and stimulation duration are used as adjustable variables. Effective parameter combinations are explored through changes in energy. Meanwhile, frequency and simulated focal plane distance are used as basic parameters and are not included in the variable exploration scope of each experiment. Instead, they are pre-set based on the group characteristics of the target population (such as the average ovarian depth corresponding to different races, body types, and animal models) and the specific symptoms of PCOS (such as ovarian volume, hormone levels, etc.), forming the basic framework for personalized treatment.

[0048] It should be understood that the parameter localization method and matching reagent kit for suppressing PCOS-related epilepsy based on focused ultrasound provided in this application have excellent generalization ability and clinical adaptability. On the one hand, after completing a parameter exploration for a specific population (such as different races and body types) or a specific PCOS subtype, the obtained dose-response curve and optimal parameter combination can be repeatedly reused in the same population, improving the clinical implementation efficiency of the treatment plan. On the other hand, when facing new target populations or new ultrasound instruments, the dose-response curve and optimal parameter combination can be quickly determined within the existing framework through a simplified experimental design, flexibly adapting to the physiological characteristics of different populations and the parameter settings of different ultrasound instruments. Thus, this application significantly simplifies the experimental process, shortens the research and development cycle, and reduces the amount of animals used and resource consumption while ensuring treatment accuracy and safety. It provides a fast, reliable, and easily promoted implementation path for the personalized clinical application of focused ultrasound in the complex scenario of PCOS-related epilepsy.

[0049] The following detailed description of some embodiments of this application is provided in conjunction with the accompanying drawings. Unless otherwise specified, the following embodiments and features can be combined with each other.

[0050] Please see Figure 1 , Figure 1 This is a schematic flowchart illustrating a parameter localization method for suppressing PCOS-related epilepsy based on focused ultrasound, as provided in an embodiment of this application. Figure 1 As shown in the embodiment of this application, a parameter localization method for suppressing PCOS-related epilepsy based on focused ultrasound is provided.

[0051] S101: Obtain multiple preset power levels, each corresponding to a different combination of power and duty cycle; based on prior experimental data and a biothermal model, predict the safe range of stimulation duration for different power levels.

[0052] Specifically, several preset levels are obtained. The preset levels refer to several pre-defined ultrasound output levels (such as level 1, level 2, level 3, etc.). Each level corresponds to a fixed combination of power and duty cycle. As the level increases, the product of power and duty cycle gradually increases, representing stronger energy output. The specific number of levels and parameters can be flexibly set according to the equipment performance and treatment needs.

[0053] Prior experimental data refers to experimental information obtained in advance, which may come from animal models (such as rats and mice), computer simulation results, or critical points of tissue thermal damage observed in human trials. By combining a small amount of prior experimental data, such as the critical stimulation duration that causes thermal damage to the ovary or surrounding tissues in a rat model at a certain intensity level, the data is input into the biothermal model to estimate the maximum safe duration at different intensity levels without causing burns or tissue damage, thus defining a safe stimulation duration range for each intensity level.

[0054] In some embodiments, the prior experimental data includes experimental data on immediate tissue damage caused by at least one intensity level at the first stimulation duration. Immediate tissue damage refers to definite tissue damage observed visually, via imaging, or through simplified pathological examination within a short period (e.g., within minutes) after the ultrasound stimulation ends, such as skin redness, swelling, or bleeding. For example, when intensity level 4 has an acoustic power of 6.5W and a duty cycle of 40%, the prior experimental data may include skin burns caused by intensity level 4 at a stimulation duration of 30 seconds.

[0055] In some embodiments, the biothermal model is the Pennes model, and its expression is:

[0056] ;

[0057] in, Sound power; Duty cycle; For stimulation duration; For the volume of the tissue acting; Tissue density; For the specific heat capacity of the tissue; Blood perfusion rate; The infusion constant; This refers to the energy conversion rate.

[0058] The Pennes biothermal model describes the relationship between tissue temperature rise and parameters such as acoustic power, duty cycle, and stimulation duration. Other model parameters, such as perfusion constant, affected tissue volume, tissue density, tissue specific heat capacity, blood perfusion rate, and energy conversion rate, can be reasonably set based on the physiological characteristics of the actual experimental subjects (e.g., soft tissue of the rat abdomen), literature data, or previous measurement results. For example, the perfusion constant can be set to 200 s.

[0059] It should be understood that a biothermal model is a computational model based on the thermodynamic properties of biological tissues. It is used to assess whether different combinations of parameters approach or exceed the temperature threshold that would cause burns or tissue damage, thereby determining the safe operating boundaries. Subsequent embodiments will use the Pennes biothermal model as an example, but in practical applications, the biothermal model can also be other existing biothermal models, such as the Weinbaum–Jiji (WJ) biothermal model, the biphasic hysteresis model, the vascular-scale explicit model, etc., and is not limited here.

[0060] In some embodiments, S101 further includes: predicting a safe threshold for stimulation duration at different levels based on prior experimental data and a biothermal model; obtaining the effective stimulation duration for each level, and determining an effective threshold for stimulation duration at different levels based on the effective stimulation duration; and obtaining a safe range for stimulation duration at different levels based on the safe threshold and the effective threshold.

[0061] Specifically, based on prior experimental data and mature biothermal models, the longest stimulation duration that will not cause thermal damage at each preset intensity level is calculated, i.e., the safety threshold. The shortest stimulation duration that can achieve a preliminary therapeutic effect at a specific intensity level is also obtained, i.e., the effective threshold. Using the safety threshold as the upper limit and the effective threshold as the lower limit, a safe stimulation duration range is defined for each intensity level. When focused ultrasound is performed at each intensity level within its corresponding safe range, the risk of tissue damage can be avoided, and the effectiveness of ultrasound intervention can be ensured.

[0062] In some embodiments, prior experimental data and the effective stimulation duration for each level can be derived from simplified pre-experiments. For example, a test stimulation duration is applied to an animal model at a certain level to observe whether immediate thermal damage (such as obvious redness and swelling) occurs in the ovary or surrounding tissues. If minor damage is confirmed, the combination of that level and duration is marked as an unsafe threshold, and the safe threshold is then calculated. In other embodiments, prior experimental data and the effective stimulation duration for each level can be derived from existing experimental data.

[0063] It should be understood that in the initial stage, using a small amount of readily available prior experimental data combined with mature biothermal models to quickly predict the safe range of stimulation duration at different levels avoids the high animal cost investment brought about by traditional large-scale pre-experiments and greatly shortens the early exploration cycle.

[0064] For example, suppose the ultrasound therapy device has 5 power levels, and the power and duty cycle settings for each power level are shown in Table 1 below.

[0065] Table 1: Power and Duty Cycle Settings for Each Gear

[0066]

[0067] A preliminary experiment was conducted on the abdominal region of rats. After shaving, ultrasound stimulation was applied to different abdominal areas of the same rat at levels 3, 4, and 5 for 30 seconds. The results showed that levels 4 and 5 induced skin burns within 30 seconds, while level 3 did not cause any damage in the same amount of time, indicating that it was near the safety margin. Given that the normal skin temperature of rats is approximately 34°C, significant thermal damage (such as second-degree burns) can occur when tissue temperature rises to 48–50°C and remains there for several minutes, and when tissue temperature reaches 55°C and remains there for about 30 seconds. Therefore, it can be inferred that the temperature rise was approximately 20°C when the burn occurred.

[0068] Based on this, using the Pennes biothermal model to compare the two burn levels at the time of burns, the ratio formula can be simplified to:

[0069] ;

[0070] Taking the prior experimental data of level 4 to estimate the safety threshold of level 3 as an example, at this time, This is the temperature rise at level 4 when a burn occurs. This refers to the temperature rise at level 3 when a burn occurs. The duty cycle is set to 4 levels (e.g., 40%). The sound power is set to 4 levels (e.g., 6.5W). The stimulation duration for burns at level 4 (e.g., 30 seconds); The duty cycle is set to level 3 (e.g., 30%). The sound power is set to level 3 (e.g., 6.5W). This represents the duration of stimulation required to cause a burn at level 3, which is the amount to be estimated.

[0071] Assuming that the temperature at both the 4th and 3rd settings increased by 20°C when the burn occurred, then Substitute the data into the ratio formula:

[0072] ;

[0073] Therefore, t2 can be estimated to be approximately 41 seconds. In other words, if a 41-second focused ultrasound is performed at level 3, theoretically, the risk of tissue damage is comparable to that of a 30-second focused ultrasound at level 4.

[0074] For safety and conservatism, the safety threshold for level 3 is set at 30 seconds. Similarly, further calculations show that the safety threshold for level 2 should be less than 72 seconds, and the safety threshold for level 1 should be less than 120 seconds. In some embodiments, when multiple safety thresholds are estimated based on different prior experimental data, the smallest safety threshold or the average of multiple safety thresholds can be selected to avoid the risk of burns.

[0075] In some embodiments, the safety thresholds estimated based on the Pennes biothermal model can also be experimentally validated. For example, for level 3, the biothermal model estimates that the risk of burn should be equivalent to that of level 4 at approximately 30 seconds, similar to level 4 at approximately 41 seconds. In independent animal experiments, applying stimulation to level 3 for a duration close to or slightly longer than this (e.g., 40–45 seconds) and observing thermal damage to the skin or subcutaneous tissue indicates that the model prediction is consistent with the actual biological effect. Such validation can be used to confirm the applicability of the Pennes biothermal model and the rationality of the parameters used under the experimental conditions (e.g., a perfusion constant of 200 s).

[0076] S102, based on the safe range of stimulation duration for each level, several combinations of experimental parameters with different preset doses are set, wherein the dose is the product of power, duty cycle and stimulation duration.

[0077] Specifically, the product of power, duty cycle, and stimulation duration is defined as a unified dose. This integrates different level parameters. Then, based on the preset dose level, an appropriate stimulation duration is selected within the safety range of each level, so that the dose calculated by the combined parameters is as close as possible to the preset dose level. For example, to achieve the preset dose of approximately 75, 30 seconds can be selected at level 3 (6.5W, 30%), approximately 70 seconds at level 2 (4W, 30%), and approximately 100 seconds at level 1 (2W, 40%), with all these stimulation durations falling within the safety range of their respective levels.

[0078] The resulting combinations of parameters, though derived from different gear levels, have similar dose values. This allows for the coverage of diverse operating conditions through limited experiments, while ensuring safety, thus providing data support for the subsequent construction and analysis of dose-effect relationships.

[0079] In some embodiments, S102 further includes: calculating the target stimulation duration of the current level based on the preset dose and the power and duty cycle corresponding to each level; if the target stimulation duration exceeds the safe range of stimulation duration under the current level, then the experimental parameter combination of the current level is not set within the preset dose.

[0080] Specifically, for each preset dose, the theoretical stimulation duration required to achieve the dose is first calculated based on the power and duty cycle of the current setting. The calculation formula is "target stimulation duration ≈ dose / (power × duty cycle)". When the target stimulation duration exceeds the safe range of the corresponding setting, the use of that setting at that dose is actively excluded to avoid the risk of tissue damage caused by excessive stimulation duration and the invalid or unstable experimental results caused by excessively low stimulation duration.

[0081] For example, to achieve a target dose of approximately 34, a stimulation duration of approximately 30 seconds is selected at level 2 (4W, 30%), and approximately 40 seconds is selected at level 1 (2W, 40%). These stimulation durations all fall within the safe range for their respective levels. The target stimulation duration for level 3 (6.5W, 30%) is calculated to be approximately 17 seconds, but this duration is below its effective threshold (e.g., 20 seconds). Therefore, this combination is considered unstable or ineffective due to its excessively low duration, and the parameter combination for level 3 is not set at this dose level. This screening mechanism ensures that all parameter combinations used satisfy both dose consistency and safety and effectiveness constraints.

[0082] In some embodiments, the preset dose can be set based on the upper limit dose and preset division parameters (such as 1, 1 / 2, 1 / 4, 3 / 2, 1 / 3, etc.). The upper limit dose refers to the maximum effective dose achievable at each level while ensuring safety; for example, a dose that does not cause burns in prior experimental data, or a dose that does not cause burns after conversion using a biothermal model. The upper limit doses are close across different levels. A series of preset doses satisfying the specified proportions can be generated according to the preset division parameters, such as upper limit dose, 1 / 2 upper limit dose, 1 / 4 upper limit dose, or, for example, upper limit dose, 3 / 2 upper limit dose, 1 / 3 upper limit dose, to cover the potential effective range from low to high.

[0083] In some embodiments, each dose level includes a high-dose group with a maximum dose and a medium-dose group with half the maximum dose. For example, the maximum dose is approximately 75, and the high-dose group may include combinations such as 3 levels for 30 seconds, 2 levels for 70 seconds, and 1 level for 100 seconds; the medium-dose group has a dose of approximately 34, and may include 2 levels for 30 seconds and 1 level for 40 seconds.

[0084] This ensures that the dosage gradient is reasonable, requiring only a small number of experiments to cover a wide parameter space, while avoiding the blind selection of parameters that exceed the safe or ineffective range. This ensures that all experimental parameter combinations are within a safe and potentially effective range, thereby improving experimental efficiency.

[0085] S103, based on several combinations of the experimental parameters, the experimental model is subjected to ultrasonic stimulation to obtain the corresponding biological response results.

[0086] Specifically, the abdominal ovarian region of the experimental model (such as a rat with PCOS and epilepsy) was irradiated once according to the experimental parameter combination. For example, approximately 60 adult female rats were prepared and divided into three phases. In each phase, the PCOS model was established by daily gavage administration of letrozole 1 mg / kg (with 1% carboxymethyl cellulose sodium as a dissolving agent) for three consecutive weeks. After successful model establishment was verified by vaginal smears, the location of the ovaries was determined and marked using ultrasound. After anesthetizing the rats with isoflurane, focused ultrasound was used to perform multiple stimulations on the ovarian region using an ultrasound therapy device.

[0087] After the experimental model is stimulated with specific ultrasound parameters, measurable physiological or behavioral changes (such as susceptibility to epilepsy) are detected as biological response results, such as seizure duration and frequency. For example, the detection method can be to induce epilepsy by intraperitoneal injection of pentylenetetrazol (PTZ) 30 mg / kg, and record the seizures as biological response results.

[0088] In some embodiments, the biological response results include: seizure time obtained based on an epilepsy susceptibility test; the ultrasound stimulation is applied every other day in step S103, alternating with the epilepsy susceptibility test. It should be understood that ovarian function recovery and ovulation in experimental models (such as rats) require time, and the anesthetic isoflurane used during ultrasound stimulation can interfere with the epilepsy susceptibility test. Therefore, for the accuracy of the results, the ultrasound stimulation is scheduled every other day, spaced apart from the epilepsy susceptibility test.

[0089] For example, on day 1, a single ultrasound stimulation is applied to a PCOS epilepsy model rat; on day 2, an epilepsy susceptibility test is performed; on day 3, ultrasound stimulation is repeated; on day 4, another epilepsy susceptibility test is performed, and so on. The experimental process does not require surgical exposure of the ovary, tissue sectioning, or ex vivo manipulation; it only involves non-invasive ultrasound irradiation through surface localization, directly reflecting the therapeutic effect. The entire process is short, simple to operate, and provides intuitive data.

[0090] Please see Figure 2 , Figure 2 This application provides an embodiment of an epileptic seizure trend graph during ultrasound stimulation. For example... Figure 2 As shown, during the first 2-3 sessions of ultrasound stimulation, there was no significant improvement in seizure frequency. After 4 sessions, the high-dose group showed that level 3 (30s) (orange line) exacerbated seizure severity, while level 2 (30s) (blue line) had a certain inhibitory effect on seizures. Correspondingly, in four consecutive epilepsy susceptibility tests on the rat model, the seizure duration at level 3 (30s) was 181%, 184.8%, 93.5%, and 250% of that in the control group, while the seizure duration at level 2 (30s) was 92.59%, 21.21%, 13.85%, and 26.79% of that in the control group.

[0091] It should be understood that, during the experimental design phase, by introducing dosage as a unified metric, a limited number of representative dosage groups are set up. Each group contains parameter combinations with different levels but similar dosages, covering a wide parameter space with a small number of experiments, thus reducing the number of experimental operations and resource consumption. Furthermore, the experimental procedure is simple and efficient: ultrasound stimulation is performed every other day, alternating with seizure time testing, eliminating the need for complex pathological sections, ex vivo organ manipulation, or invasive testing, allowing for rapid and intuitive data acquisition, further shortening the experimental cycle and lowering the technical threshold and cost.

[0092] S104, based on the preset dose-response relationship, fits the dose-response curves for each level according to the biological response results of each experimental parameter combination at each level.

[0093] Specifically, based on a pre-assumed dose-response relationship (such as a single-peak curve), the dose values ​​corresponding to different parameter combinations at each level and their biological response results are correlated and fitted. For example, based on the fact that level 2 obtains significant and moderate onset delay at 30 seconds and 70 seconds, respectively, combined with the pre-defined single-peak trend, the peak effect at this level can be quickly interpolated to determine that it may be located in the dose range of 34–75.

[0094] It should be understood that although the fitting curves sacrifice some absolute accuracy due to limited experimental data, they can effectively reflect the overall trend of change, quickly determine the potential high-efficiency dose range, and provide a basis for decision-making on whether to add a validation group in this range.

[0095] In some embodiments, the preset dose-response relationship includes one or more of the following:

[0096] a. The dose-response curve exhibits a unimodal distribution within the safety parameter range;

[0097] b. The peak dose of the dose-response curve is inversely correlated with the power and duty cycle values ​​in the gear setting;

[0098] c. The absolute value of the slope of the dose-response curve is positively correlated with the values ​​of power and duty cycle in the gear.

[0099] d. The maximum rate of change of curvature of the dose-response curve is less than the preset rate of change threshold.

[0100] The preset dose-response relationship 'a' is set based on Hormesis theory. It should be understood that biological systems generally exhibit a biphasic dose-response relationship to external stimuli, that is, lower doses produce stimulating / beneficial effects, while higher doses produce inhibitory / harmful effects, thus forming an inverted U-shaped single-peak dose-response curve. This biphasic dose-response curve has been widely observed, and most drugs conform to this phenomenon (i.e., there is only one recommended dose). This phenomenon is applied to ultrasound therapy.

[0101] The preset dose-effect relationships b and c are set based on the fundamental principles of ultrasound biological effects. Ultrasound effects can be divided into thermal and mechanical effects. Within common parameter ranges, both thermal and mechanical effects increase approximately exponentially with increasing ultrasound intensity (defined as the product of power and duty cycle), while the dose (defined as the product of power, duty cycle, and stimulation duration) increases linearly with ultrasound intensity. It should be understood that at the same dose, higher intensity levels result in higher energy utilization and a faster peak response; while lower intensity levels require longer durations (i.e., higher doses) to accumulate sufficient effects.

[0102] Therefore, the dose-response curve of a higher dose level typically reaches its peak at a lower dose, and the rise / fall is steeper (the absolute value of the slope is larger); conversely, the peak of a lower dose level curve appears at a higher dose, and the change is gentler (the absolute value of the slope is smaller). For example, dose level 2 may achieve the best anti-epileptic effect at a dose of 34, while dose level 1 requires a dose of 70 or higher to approach the same effect.

[0103] The preset dose-effect relationship d is set based on the constraint of curve smoothness to avoid unreasonable fluctuations caused by sparse experimental data or noise interference. Among them, the maximum rate of change of curvature refers to the maximum value of the curvature (i.e., the rate of change of the curvature) on the dose-effect curve. Correspondingly, the preset rate of change threshold is set based on the reasonable dynamic range in actual application to ensure that the fitted curve presents a gradual and continuous trend change.

[0104] In some embodiments, please refer to Figure 3 , Figure 3 This is a schematic diagram of dose-response curves at various levels provided in the embodiments of this application, such as... Figure 3 As shown, the horizontal axis of the dose-response curve represents the ultrasound dose, and the vertical axis represents the treatment effect. The treatment effect can be the average of the results of each epilepsy susceptibility test. The treatment effect can be specifically quantified as the average of the results of multiple epilepsy susceptibility tests, such as the average seizure time of each epilepsy susceptibility test in the rat models of each experimental group; other physiological indicators, such as the severity of epilepsy seizures and the number of seizures per unit time, can also be used.

[0105] In some embodiments, the therapeutic effect is defined as the average difference in the duration of grade 5 epileptic seizures between the control group and the experimental group across all rounds. For example, an epilepsy susceptibility test is performed after each ultrasound stimulation, and the number of epilepsy susceptibility tests is [number missing]. Each test included a PCOS control group and several experimental groups receiving specific ultrasound parameters. The average seizure time across the experimental groups was [value missing]. The mean seizure time in the PCOS control group was The formula for calculating the treatment effect for each epilepsy susceptibility test is as follows: .

[0106] Taking level 2, 70 seconds as an example, the treatment effect can be quantified and calculated using data from four tests:

[0107] .

[0108] Thus, the therapeutic effects of different dose levels at different intensity levels were quantified, and biological response data points at each intensity level were obtained (e.g., ... Figure 3 (The solid dots in the data) represent the therapeutic effect of a specific dose at a certain level. Based on these limited data points, curve fitting is performed in combination with the preset dose-effect relationship to deduce the dose-effect trend of each level.

[0109] In some embodiments, based on a preset dose-effect relationship and the obtained biological response data points for each gear level, the predicted region of the peak dose for each gear level can be determined according to the therapeutic effect trend corresponding to the data points. When a gear level has missing data points due to burn risk, unreliable parameters, or experimental abnormalities, making it impossible to directly fit its peak position, the peak dose of the gear level with missing data points can be reasonably inferred based on the preset dose-effect relationship b (i.e., the peak dose of the dose-effect curve is inversely correlated with the power and duty cycle values ​​in the gear level) and the predicted regions of the peak doses of other gear levels. For example, if the predicted peak dose of the high gear level is located in a lower dose range, while the predicted peak dose of the low gear level is located in a higher dose range, the peak dose of the intermediate gear level can be interpolated accordingly.

[0110] In some embodiments, when constructing the dose-response curve, c (i.e., the absolute value of the slope of the dose-response curve is positively correlated with the values ​​of power and duty cycle in the gear) can be used to optimize the curve shape in the prediction area. For example, the high gear curve shows a steeper rising / falling trend, while the low gear curve shows a gentler trend.

[0111] like Figure 3 As shown, level 2 contains two valid data points: medium and high doses. The therapeutic effect at the high dose is not significantly better than at the medium dose, indicating that its dose-response curve may have approached or exceeded the peak value. Therefore, it is reasonable to infer that the peak value is near the medium dose. Similarly, it can be determined that the peak dose of level 1 is greater than or equal to the high dose. For level 3, which has only one valid data point due to missing data, we can infer that the peak dose of level 3 may be lower than the medium dose based on the inverse correlation between the peak dose and the power and duty cycle values ​​of the level. Combined with the fact that the absolute value of the slope of the level 3 curve is greater than that of levels 1 and 2, we can finally plot the possible dose-response curve for level 3.

[0112] Therefore, even with sparse or partially missing data, reasonable dose-response curves that conform to physical mechanisms and biological response laws can be generated, providing a reliable basis for subsequent parameter optimization.

[0113] S105, Based on the dose-effect curve, determine the preferred dose range for each level, and conduct a feasibility assessment of the preferred dose range for each level to determine the preferred parameter combination for focused ultrasound.

[0114] The optimal dose range was identified from the dose-response curves of each intensity level. Then, a comprehensive screening was conducted based on the actual feasibility of each level. Feasibility assessment could include the following dimensions: whether the optimal dose range carries a risk of immediate or delayed tissue thermal damage; whether the optimal dose range falls within the output capability of existing ultrasound equipment; and whether the optimal dose range has potential for widespread adoption. For example, a medium-intensity level is more likely to be promoted than an extremely high or low level. The priority of level promotion can be determined in advance through surveys and experiments. For instance, while level 1 is effective in the high-dose range, the required stimulation duration is close to or exceeds its safety threshold, posing a risk of burns. Level 3 has an optimal dose in the low-to-medium dose range, but due to its excessively high power and duty cycle, it still carries a risk of burns in practical applications. In contrast, the optimal dose range of level 2 falls entirely within its safety range, and the experimental data is stable and reproducible. Therefore, level 2 and its corresponding parameters (e.g., level 2 for 30 seconds) were ultimately selected as the optimal parameter combination.

[0115] It should be understood that the optimal parameter combination is not theoretically optimal, but rather a feasible scheme that has been experimentally verified as suitable and effective. It can be used for subsequent animal replication and validation, adaptation to different PCOS subtypes, or as an initial parameter template for clinical translation.

[0116] In some embodiments, the preferred parameter combination is retested to verify parameter stability. For example, an additional repeat test was performed on a group (e.g., 5 rat models) of treatment at level 2 for 30 seconds, and the results showed that the average duration of attacks at level 2 for 30 seconds was approximately 35% of that in the PCOS control group, indicating that the parameters were stable.

[0117] In some embodiments, the number of ultrasound stimulations is 8 to 10. The therapeutic effect is significant and relatively stable after two weeks with a dose of 30 seconds at level 2, therefore the stimulation duration can be 10 times.

[0118] In some embodiments, the method further includes: during the execution of step S103, monitoring whether delayed tissue damage occurs; if so, updating the safety range of the corresponding gear based on the experimental data of the delayed tissue damage and the biothermal model.

[0119] Specifically, during ultrasound stimulation, the experimental animals were simultaneously observed for delayed tissue damage. For example, in the high-dose group, no abnormalities were observed on the skin surface after the first 2–3 ultrasound stimulations every other day, but burns gradually appeared after the 4th stimulation. Subsequent tissue sampling also revealed signs of burns in subcutaneous or abdominal organs, suggesting delayed damage caused by cumulative thermal effects. In this case, the total number of stimulations resulting in delayed damage at that level was combined with the duration of each stimulation, considered as a new critical condition, and estimated using the aforementioned biothermal model (such as the ratio formula based on the Pennes model). For example, if damage occurred at level 3 after a total of 4 stimulations and a single stimulation duration of 30 seconds, it can be equivalently deduced that level 2 might trigger a similar delayed risk at a stimulation duration of 50 seconds, thus adjusting the upper limit of the safety range for level 2 from the original 72 seconds to 50 seconds. This updated safety range can be used to guide parameter adjustments or secondary fine-tuning in subsequent experiments, improving experimental safety.

[0120] In some embodiments, the magnitude and trend of the biological response to half the upper limit dose and the upper limit dose meet the following typical pattern, allowing for the direct construction of a reasonable dose-response curve based on limited data: the lower dose group monotonically increases within the upper limit dose range, the higher dose group begins to decrease before the half upper limit dose, and the optimal value for the relative middle dose group lies between the upper and lower doses. If there are many dose levels (e.g., more than 5), the curve shape of adjacent dose levels can quickly converge to a narrower preferred dose range; conversely, if there are few dose levels (e.g., only 3), it is necessary to combine the existing curve shape to infer the possible optimal peak range, and supplement parameters within the possible peak range for secondary verification.

[0121] For example, the preferred dose range includes the peak position of the dose-effect curve, and S105 includes: when the dose-effect curve has multiple candidate peaks and / or the preferred dose range is greater than a preset range threshold, obtaining a target experimental parameter combination within the preferred dose range; setting at least two sets of verification parameter combinations at the same level of the target experimental parameter combination; wherein, the dose of one set of verification parameter combinations is greater than the dose of the target experimental parameter combination, and the dose of the other set of verification parameter combinations is less than the dose of the target experimental parameter combination; executing steps S103 to S105 to update the dose-effect curve based on the data of the verification parameter combinations.

[0122] Specifically, if the dose-response curve for a certain dose level has multiple candidate peaks or the preferred dose range is too wide due to data sparsity, a refined verification process is initiated. For example, in the aforementioned example, it is speculated that the peak dose of dose level 2 is near the medium dose, but it is impossible to clearly determine whether the peak dose of dose level 2 is located at the medium dose, below the medium dose, or above the medium dose. At this time, there may be multiple candidate peaks, making the prediction of the dose curve inaccurate or requiring the division of a larger preferred dose range.

[0123] Obtain a target experimental parameter combination within the preferred dose range (e.g., level 2, 30 seconds, dose 36) as a baseline, and add two sets of validation parameter combinations at the same level: one set with a slightly higher dose (e.g., level 2, 40 seconds, dose 48), and the other set with a slightly lower dose (e.g., level 2, 20 seconds, dose 24), both falling within the safety range of level 2. Then repeat steps S103 to S105, obtain the newly added biological response data, and refit the dose-response curve.

[0124] Please see Figures 4 to 5 , Figure 4 This is a schematic diagram of another dose-response curve at various levels provided in the embodiments of this application. Figure 5 This is a schematic diagram of dose-response curves at various levels provided in an embodiment of this application. In some cases, such as Figure 4 As shown, the blue hollow dots represent newly added validation data points. The curve trend indicates that the peak value is biased towards the higher dose side. Based on this, the optimal dose range of level 2 can be corrected, and the validation parameter combination for higher doses (such as level 2 for 40 seconds) can be updated to the new optimal parameter combination. In other cases, such as Figure 5 As shown, the yellow hollow dots represent newly added validation data points, and the curve trend indicates that the peak value is biased towards the low-dose side. Figure 3 If the initial prediction of the peak position is incorrect, the preferred dose range should be adjusted to a lower dose, and a lower dose combination of validation parameters (such as 2 levels for 20 seconds) should be adopted as a better choice. This allows for a quick and easy refinement of the parameters, providing a better parameter range foundation for subsequent personalized medicine.

[0125] In some embodiments, after the step of obtaining multiple preset levels, the method further includes: adjusting the preset division parameter of the preset dose according to the number of preset levels to update the value and quantity of the preset dose; wherein, the fewer the number of preset levels, the more preset doses are corresponding to the preset division parameter.

[0126] For example, when the number of preset dose levels is small, the preset division parameter of the preset dose can be adjusted to three equal parts: the upper limit dose, 3 / 2 of the upper limit dose, and 1 / 3 of the upper limit dose. This improves the discriminative power of the initial dose-response curve, thereby more clearly predicting the dose-response trend at each dose level and avoiding peak misjudgment due to insufficient dose coverage. As another example, when the number of preset dose levels is large, the preset division parameter of the preset dose can be adjusted to two equal parts: the upper limit dose and 1 / 2 of the upper limit dose. Although the data points for each dose level are fewer, the dose-response curve at each dose level can be predicted by combining the preset dose-response relationship and the curve shapes of multiple dose levels.

[0127] In some embodiments, changes in the body weight of the animal model are detected during the experiment. See also Figure 6 , Figure 6 This is a graph showing the weight changes of the animal model provided in this application at key time points, such as... Figure 6 As shown, the experiment included a PCOS control group and three dosage experimental groups: a high-dose group, a medium-dose group, and a low-dose group. After letrozole administration, the body weight of the experimental models (e.g., rats) began to increase. After the ultrasound was initiated, the body weight decreased, and then gradually recovered after the ultrasound ended. Rapid weight gain is a characteristic of PCOS. The slower rate of weight gain after ultrasound may indicate a therapeutic effect of ultrasound stimulation, but other factors such as inflammation, reduced food intake, or gastrointestinal irritation cannot be ruled out.

[0128] In some embodiments, after the test is completed, perfusion sampling is performed, serum is subjected to mass spectrometry hormone detection, and brain tissue is subjected to immunofluorescence staining and mass spectrometry analysis, etc. Please refer to Figure 7 , Figure 7 This is a comparison chart of serum hormone levels between different experimental groups provided in the embodiments of this application, such as... Figure 7 As shown, mass spectrometry analysis was performed on the hormones in each group. The results showed that all parameters could increase the estrogen level in PCOS rats. The estrogen level in ordinary PCOS rats was about 20% of the normal level. The estrogen level in the medium-dose group (e.g., level 2 for 30 seconds) could increase to 3.5 times that of the PCOS group, which is close to 80% of the normal rat level. The other parameters could also be increased to about 50% of the normal rat level. The androgen level in the medium-dose group was about 20% lower than that in the PCOS group. The other parameters could not reduce the androgen level, but instead showed an increasing trend.

[0129] In some embodiments, when the experimental model is a rat, the preferred parameter combination includes: acoustic power of 4W, duty cycle of 30%, stimulation duration of 30s, frequency of 1.028MHz, and simulated focal plane distance of 11.5mm. Rats stimulated with these parameters experienced approximately 30% of the duration of grade 5 seizures in the PCOS control group, suggesting a significant reduction in the burden of severe seizures.

[0130] This application also provides a reagent kit 200, please refer to... Figure 8 , Figure 8 This is a schematic block diagram of a reagent kit provided in an embodiment of this application. The reagent kit 200 can be applied to an experimental setup to implement the aforementioned parameter localization method for suppressing PCOS-related epilepsy based on focused ultrasound.

[0131] like Figure 8 As shown, the kit 200 includes:

[0132] The safe interval testing module 201 is used to acquire multiple preset levels, each level corresponding to different combinations of power and duty cycle; based on prior experimental data and biothermal models, it predicts the safe interval of stimulation duration at different levels.

[0133] The experimental parameter setting module 202 is used to set several combinations of experimental parameters with different preset doses according to the safe range of stimulation duration for each level. The dose is the product of power, duty cycle and stimulation duration.

[0134] The ultrasound stimulation execution module 203 is used to perform ultrasound stimulation on the experimental model based on several combinations of the experimental parameters, and obtain the corresponding biological response results.

[0135] The dose-response curve module 204 is used to fit the dose-response curve for each level based on a preset dose-response relationship and the biological response results of each combination of experimental parameters at each level. The preset dose-response relationship includes: a. the dose-response curve has a single peak distribution within the safe parameter range; b. the peak dose of the dose-response curve is inversely correlated with the power and duty cycle values ​​in the level.

[0136] The preferred dose range module 205 is used to determine the preferred dose range for each level based on the dose-effect curve, and to conduct a feasibility assessment of the preferred dose range for each level in order to determine the preferred parameter combination for focused ultrasound.

[0137] For example, the prior experimental data includes experimental data on immediate tissue damage caused by at least one setting at the first stimulation duration;

[0138] The expression for the biothermal model is:

[0139] ;

[0140] in, Sound power; Duty cycle; For stimulation duration; For the volume of the tissue acting; Tissue density; For the specific heat capacity of the tissue; Blood perfusion rate; The infusion constant; This refers to the energy conversion rate.

[0141] For example, the safety interval test module 201 also includes a safety threshold submodule, an effective threshold submodule, and a safety interval submodule;

[0142] The safety threshold submodule is used to predict the safety threshold of stimulation duration at different levels based on prior experimental data and biothermal models.

[0143] The effective threshold submodule is used to obtain the effective stimulation duration for each level and determine the effective threshold of stimulation duration for different levels based on the effective stimulation duration.

[0144] The safety interval submodule is used to obtain the safety interval of stimulation duration at different levels based on the safety threshold and the effective threshold.

[0145] For example, the experimental parameter setting module 202 also includes: a target duration submodule and a parameter combination submodule;

[0146] The target duration submodule is used to calculate the target stimulation duration of the current level based on the preset dose and the power and duty cycle corresponding to each level.

[0147] The parameter combination submodule is used to not set the experimental parameter combination for the current level within a preset dose range if the target stimulation duration exceeds the safe range of stimulation duration at the current level.

[0148] For example, the kit 200 also includes a delayed injury detection module for monitoring whether delayed tissue injury occurs; if so, the safety range of the corresponding level is updated based on the experimental data of the delayed tissue injury and the biothermal model.

[0149] For example, the preset dose-effect relationship also includes: c. The absolute value of the slope of the dose-effect curve is positively correlated with the values ​​of power and duty cycle in the gear; d. The maximum rate of change of curvature of the dose-effect curve is less than a preset rate of change threshold.

[0150] For example, the biological response results include: seizure time based on epilepsy susceptibility testing; the ultrasound stimulation is applied at a frequency of once every other day and is performed alternately with epilepsy susceptibility testing.

[0151] For example, the preferred dose range includes the peak position of the dose-effect curve, and the kit 200 further includes: a baseline parameter module, a validation parameter module, and a data update module;

[0152] The baseline parameter module is used to obtain a target experimental parameter combination within the preferred dose range when the dose-effect curve has multiple candidate peaks or the preferred dose range is greater than a preset range threshold.

[0153] The verification parameter module is used to set at least two sets of verification parameter combinations at the same level of the target experimental parameter combination; wherein, the dose of one set of verification parameter combinations is greater than the dose of the target experimental parameter combination, and the dose of the other set of verification parameter combinations is less than the dose of the target experimental parameter combination.

[0154] The data update module is used to trigger the ultrasound stimulation execution module 203, the dose-response curve module 204, and the preferred dose range module 205 to update the dose-response curve based on the data of the verification parameter combination.

[0155] For example, the parameters of the focused ultrasound also include frequency and simulated focal plane distance, which are determined based on the average ovarian depth of the population and the symptoms of polycystic ovary syndrome.

[0156] For example, the reagent kit 200 further includes: a division parameter adjustment module, used to adjust the preset division parameter of the preset dose according to the number of preset levels, so as to update the value and quantity of the preset dose; wherein, the fewer the number of preset levels, the more preset doses corresponding to the preset division parameter.

[0157] In some embodiments, the experimental model is a rat model of polycystic ovary syndrome; the experimental device can be used to perform ultrasound stimulation on the rat model of polycystic ovary syndrome.

[0158] It should be noted that those skilled in the art will understand that, for the sake of convenience and brevity, the specific working processes of the reagent kit and each module and unit described above can be referred to the corresponding processes in the foregoing method embodiments, and will not be repeated here.

[0159] The above description is merely a specific embodiment of this application, but the scope of protection of this application is not limited thereto. Any person skilled in the art can easily conceive of various equivalent modifications or substitutions within the technical scope disclosed in this application, and these modifications or substitutions should all be covered within the scope of protection of this application. Therefore, the scope of protection of this application should be determined by the scope of the claims.

Claims

1. A reagent kit, characterized in that, The kit is used in an experimental setup to implement a parameter localization method for suppressing PCOS-related epilepsy based on focused ultrasound. The parameters of the focused ultrasound include power, duty cycle, and stimulation duration. The kit comprises: The safe zone testing module is used to acquire multiple preset power levels, each corresponding to a different combination of power and duty cycle; based on prior experimental data and biothermal models, it predicts the safe zone of stimulation duration at different power levels. The experimental parameter setting module is used to set several combinations of experimental parameters with different preset doses according to the safe range of stimulation duration for each level. The dose is the product of power, duty cycle and stimulation duration. The fewer the preset levels, the more preset doses there are. An ultrasound stimulation execution module is used to perform ultrasound stimulation on the experimental model based on several combinations of the experimental parameters, and obtain the corresponding biological response results. The dose-response curve module is used to fit dose-response curves for each level based on a preset dose-response relationship and the biological response results of various experimental parameter combinations at each level. The preset dose-response relationship includes: a. the dose-response curve exhibits a unimodal distribution within the safety parameter range; b. the peak dose of the dose-response curve is inversely correlated with the power and duty cycle values ​​at each level; c. the absolute value of the slope of the dose-response curve is positively correlated with the power and duty cycle values ​​at each level. The preferred dose range module is used to determine the preferred dose range for each level based on the dose-effect curve, and to conduct a feasibility assessment of the preferred dose range for each level in order to determine the preferred parameter combination for focused ultrasound.

2. The reagent kit according to claim 1, characterized in that, The prior experimental data includes experimental data on immediate tissue damage caused by at least one setting at the first stimulation duration; The expression for the biothermal model is: ； in, Sound power; Duty cycle; For stimulation duration; For the volume of the tissue acting; Tissue density; For the specific heat capacity of the tissue; Blood perfusion rate; The infusion constant; This refers to the energy conversion rate.

3. The kit according to claim 1 or 2, characterized in that, The safe zone testing module also includes: The safety threshold submodule is used to predict the safety threshold of stimulation duration at different levels based on prior experimental data and biothermal models. The effective threshold submodule is used to obtain the effective stimulation duration for each level and determine the effective threshold of stimulation duration for different levels based on the effective stimulation duration. The safety interval submodule is used to obtain the safety interval of stimulation duration at different levels based on the safety threshold and the effective threshold.

4. The reagent kit according to claim 1, characterized in that, The experimental parameter setting module also includes: The target duration submodule is used to calculate the target stimulation duration of the current level based on the preset dose and the power and duty cycle corresponding to each level. The parameter combination submodule is used to not set the experimental parameter combination for the current level within a preset dose range if the target stimulation duration exceeds the safe range of stimulation duration at the current level.

5. The reagent kit according to claim 1, characterized in that, The kit also includes: The delayed damage detection module is used to monitor whether delayed tissue damage occurs during the execution of step S103; if so, it updates the safety range of the corresponding gear according to the experimental data of the delayed tissue damage and the biothermal model.

6. The reagent kit according to claim 1, characterized in that, The preset dose-response relationship also includes: d. The maximum rate of change of curvature of the dose-response curve is less than the preset rate of change threshold.

7. The kit according to claim 1, characterized in that, The biological response results include: the duration of epileptic seizures obtained based on epilepsy susceptibility testing; The ultrasound stimulation is applied in step S103 at a frequency of once every other day and is performed alternately with the epilepsy susceptibility test.

8. The reagent kit according to claim 1, characterized in that, The preferred dose range includes the peak position of the dose-response curve, and the preferred dose range module further includes: The baseline parameter module is used to obtain a target experimental parameter combination within the preferred dose range when the dose-effect curve has multiple candidate peaks or the preferred dose range is greater than a preset range threshold. The verification parameter module is used to set at least two sets of verification parameter combinations at the same level of the target experimental parameter combination; wherein, the dose of one set of verification parameter combinations is greater than the dose of the target experimental parameter combination, and the dose of the other set of verification parameter combinations is less than the dose of the target experimental parameter combination. The data update module is used to trigger the ultrasound stimulation execution module, the dose-response curve module, and the preferred dose range module to update the dose-response curve based on the data of the verification parameter combination.

9. The reagent kit according to claim 1, characterized in that, The parameters of the focused ultrasound also include frequency and simulated focal plane distance, which are determined based on the average ovarian depth of the population and the symptoms of polycystic ovary syndrome.

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