System for ablating tumor tissue based on ultrahigh-frequency extremely-short pulse electric field and application of system

Abstract

The invention belongs to the technical field of bioelectricity medicine and tissue ablation, and particularly relates to a tumor tissue ablation system based on an ultrahigh-frequency extremely-short pulse electric field and application of the tumor tissue ablation system. The system comprises a data acquisition and analysis module, a control module, a pulse generation module and an ablation module. The data acquisition and analysis module, the control module, the pulse generation module and the ablation module are connected in sequence. Wherein the data acquisition and analysis module is preferably selected from a multi-mode image navigation system, and the control module is preferably selected from a man-machine interaction control terminal. By fusing ultrahigh-frequency extremely-short pulse ablation and dynamic impedance matching regulation and control, the technical problem that ablation treatment and blood brain barrier opening are difficult to realize synchronously in the prior art is solved, and the method has important significance for realizing more accurate and safer brain tumor treatment.

Description

Technical Field

[0001] This invention belongs to the field of bioelectric medicine and tissue ablation technology, specifically relating to a system for ablating tumor tissue based on an ultra-high frequency extremely short pulse electric field and its application. Background Technology

[0002] Tissue ablation is a minimally invasive interventional treatment method that, guided by modern imaging technologies such as ultrasound, CT, and MRI, uses chemical or physical methods to cause irreversible damage or necrosis of diseased tissue cells, thereby achieving the therapeutic goal. Compared with traditional methods such as surgery, radiotherapy, and chemotherapy, tissue ablation has advantages such as high efficiency and safety, wider indications, less trauma, fewer complications, and repeatability, which can effectively prolong patients' survival and improve their quality of life.

[0003] Ablation techniques are mainly divided into two categories: thermal ablation and non-thermal ablation. Thermal ablation uses temperature changes to kill cancer cells. Traditional thermal ablation techniques such as radiofrequency ablation, microwave ablation, and laser ablation rely on heat energy conduction to destroy target tissue. Although they can achieve local ablation, they are limited by the heat diffusion effect and are prone to causing irreversible damage to surrounding healthy tissues (such as nerves and blood vessels). Furthermore, tissue carbonization may affect the clarity of the ablation boundary.

[0004] Non-thermal ablation techniques mainly include chemical ablation and pulsed electric field ablation (PFA). PFA, as a representative of non-thermal ablation techniques, induces cell membrane electroporation through high-intensity electric pulses, theoretically avoiding thermal damage and achieving selective subcellular destruction. Current technologies primarily employ microsecond-level pulses (10-100 μs) combined with a field strength of 0.5-3 kV / cm, such as irreversible electroporation (IRE). While IRE can preserve the extracellular matrix and vascular structure, its low-frequency characteristics (<1 kHz) make the electric field distribution susceptible to tissue heterogeneity, resulting in ablation boundary errors >2 mm and potentially causing side effects such as muscle contraction. High-frequency irreversible electroporation (H-FIRE) uses higher-frequency pulse sequences (such as 10 kHz biphasic pulses), which reduces neuromuscular stimulation, but still suffers from the inherent thermal accumulation effect of microsecond-level pulses (temperature rise >8°C per treatment), affecting safety. Furthermore, although existing high-frequency pulse ablation systems use 100kHz pulses, their effective high-frequency components (>50MHz) account for less than 30%, limiting their selective ablation capability for organelles. Meanwhile, short-pulse electric field devices (such as 500ns pulse width) lack dynamic impedance matching mechanisms, making it difficult to maintain a stable ultra-high frequency (>1MHz) electric field output in complex tissues, resulting in unsatisfactory ablation depth and consistency.

[0005] For deep tumors such as gliomas, the blood-brain barrier (BBB) ​​remains a significant challenge. Current mainstream methods for opening the BBB include ultrasound combined with microbubbles and microsecond pulsed electric fields. Ultrasound combined with microbubbles utilizes focused ultrasound (0.5-2 MHz) combined with the cavitation effect of microbubbles (1-10 μm) to mechanically disrupt the tight junctions of the BBB, achieving reversible opening within 4-6 hours. However, relying on microbubble injection can easily cause microvascular damage, and the depth of ultrasound penetration is limited by skull attenuation. Microsecond pulsed electric field technology (100-500 μs pulse width, 0.1-1 kV / cm field strength) temporarily disrupts the BBB through electroporation, eliminating the need for microbubbles, but the opening time is short (<6 hours), and the low-frequency electric field is susceptible to tissue heterogeneity, leading to uneven opening areas.

[0006] In summary, current technologies struggle to simultaneously achieve ablation therapy and blood-brain barrier opening, resulting in poor spatiotemporal synergy between drug delivery and tumor clearance, thus impacting overall therapeutic efficacy. Therefore, there is an urgent need for an innovative technology that can simultaneously achieve high-precision tumor tissue ablation and controllable BBB opening to improve the treatment outcomes for central nervous system diseases such as gliomas. Summary of the Invention

[0007] In view of this, to overcome the problems of thermal damage risk, insufficient ablation precision, and uneven electric field distribution in existing ablation technologies, as well as the challenges of BBB opening technology such as microbubble dependence, short opening time, and insufficient penetration depth, this invention proposes a system for ablating tumor tissue based on ultra-high frequency, extremely short pulse electric field, and its application. This system can achieve tissue ablation of deep tumors such as gliomas, while selectively opening the blood-brain barrier, which provides support for the treatment of gliomas.

[0008] One of the objectives of this invention is to provide a system for ablating tumor tissue based on an ultra-high frequency, extremely short pulse electric field.

[0009] To achieve the above objectives, the present invention adopts the following technical solution:

[0010] A system based on ultra-high frequency, extremely short pulse electric field ablation of tumor tissue, the system comprising:

[0011] Data acquisition and analysis module: used to acquire and analyze MRI, CT, and ultrasound test results to obtain the location and size of the ablation target area and the electrode implantation location;

[0012] Control module: The control module is connected to the data acquisition and analysis module to generate treatment pulse parameters based on the image detection results of the data acquisition and analysis module;

[0013] Pulse generating module: The control module is connected to the pulse generating module to control the pulse generating module to output pulses with set parameters or to cut off the pulse electric field;

[0014] Ablation module: It is equipped with a minimally invasive single-needle electrode and a flat grounded electrode; the pulse generation module is connected to the minimally invasive single-needle electrode and the flat grounded electrode respectively through wires; the pulse generation module outputs a pulsed electric field and targets and ablates tumor tissue through the ablation module.

[0015] Preferably, the pulse parameters include pulse amplitude, polarity, pulse width, and frequency.

[0016] Preferably, the pulse generation module is used to output a unipolar or bipolar high-voltage pulse with a pulse width of 10-100ns, a frequency of 0.5-10MHz, and an amplitude of 0-±5kV.

[0017] Preferably, the pulse generation module is used to output a pulsed electric field with a pulse width of 10-100ns, a frequency of 0.5-10MHz, and a value of unipolar 0-5kV or bipolar 0-±5kV.

[0018] Preferably, the pulse generating module outputs a pulse electric field with the following parameters: pulse amplitude of 1-5kV, unipolar or bipolar, frequency within the pulse train of 0.5-10MHz, and single pulse width of 10-100ns.

[0019] Preferably, the number of pulses is 20-100.

[0020] As a preferred embodiment, the pulsed electric field has an electric field strength of 50 kV / cm (i.e., pulse amplitude of 5 kV and electrode spacing of 1 mm), a single pulse width of 50 ns, a frequency of 2 MHz within the pulse train, is bipolar, and consists of 20 pulsed electric fields. This parameter configuration can effectively target and inhibit glioma activity.

[0021] As a preferred embodiment, the pulsed electric field is a unipolar pulsed electric field with an electric field strength of 30 kV / cm (i.e., pulse amplitude of 3 kV and electrode spacing of 1 mm), a pulse width of 50 ns, and a frequency of 2 MHz. This parameter configuration can effectively inhibit the division and migration abilities of tumor cells, leading to immunogenic death of tumor cells.

[0022] As a preferred embodiment, the pulsed electric field is a unipolar pulsed electric field with an electric field strength of 20 kV / cm (i.e., pulse amplitude of 2 kV and electrode spacing of 1 mm), a pulse width of 50 ns, and a frequency of 2 MHz. This parameter configuration can induce DNA damage in glioma cells.

[0023] As a preferred embodiment, the pulsed electric field is a bipolar pulsed electric field with a field strength of 20 kV / cm (i.e., pulse amplitude of 2 kV and electrode spacing of 1 mm), a pulse width of 50 ns, and a frequency of 2 MHz. Treatment with this pulsed electric field configuration resulted in an increase in intracellular dsDNA.

[0024] As a preferred embodiment, the pulsed electric field is a bipolar pulsed electric field with a pulse amplitude of 3kV, a pulse width of 50ns, and a frequency of 2MHz. This parameter configuration can effectively inhibit the development of gliomas in situ.

[0025] As a preferred embodiment, the pulsed electric field is a bipolar high-frequency extremely short pulsed electric field with a pulse width of 50 ns, an amplitude of 3 kV, an intra-series frequency of 2 MHz, an intra-series pulse count of 100, and a pulse train of 100 pulses. Under this parameter configuration, the permeability of the blood-brain barrier is significantly improved.

[0026] Preferably, the flat grounding electrode is a flat grounding copper foil electrode and / or a PEDOT:PSS conductive polymer patch.

[0027] Preferably, the pulse generation module includes a pulse generator and a pulse output switch; the pulse output switch is connected between the pulse generator and the ablation module to control the on / off state of the pulse electric field.

[0028] Preferably, the minimally invasive single-needle electrode is implanted in the center of the tumor tissue, and the flat ground electrode is attached to the skin surface to form a closed circuit.

[0029] Preferably, the data acquisition and analysis module is a multimodal image navigation system.

[0030] Preferably, the control module is a human-computer interaction control terminal.

[0031] Preferably, the pulse generator is a solid-state pulse generator.

[0032] The second objective of this invention is to provide the aforementioned system and / or pulsed electric field-based ablation of tumor tissue using ultra-high frequency extremely short pulsed electric fields in the preparation of products for ablation of glioma tissue, enhancing blood-brain barrier permeability and / or inhibiting glioma development.

[0033] To achieve the above objectives, the present invention adopts the following technical solution:

[0034] The aforementioned system and / or pulsed electric field are used in the preparation of products for ablating glioma tissue, increasing blood-brain barrier permeability, and / or inhibiting glioma development.

[0035] Preferably, the pulsed electric field is a unipolar or bipolar high-voltage electric pulse with a pulse width of 10-100ns, a frequency of 0.5-10MHz, and an amplitude of 0-±5kV.

[0036] Preferably, the pulsed electric field is a pulsed electric field with a pulse width of 10-100ns, a frequency of 0.5-10MHz, and a value of unipolar 0-5kV or bipolar 0-±5kV.

[0037] Preferably, the products include, but are not limited to, medical devices, pharmaceutical compositions, and fields.

[0038] As a preferred embodiment, the aforementioned system is used in the preparation of a device for ablating glioma tissue, enhancing blood-brain barrier permeability, and / or inhibiting glioma development; or the pulsed electric field is used in the preparation of a field and / or pharmaceutical composition for ablating glioma tissue, enhancing blood-brain barrier permeability, and / or inhibiting glioma development.

[0039] The third objective of this invention is to provide an application of the aforementioned system based on ultra-high frequency, extremely short pulse electric field ablation of tumor tissue in the preparation of products for killing glioma cells, targeting and inhibiting glioma activity, inhibiting glioma cell division and migration, inhibiting tight junction protein ZO-1 expression, and / or reducing TEER.

[0040] To achieve the above objectives, the present invention adopts the following technical solution:

[0041] The aforementioned system is used in the preparation of products for killing glioma cells, targeting and inhibiting glioma activity, inhibiting glioma cell division and migration, inhibiting tight junction protein ZO-1 expression and / or reducing TEER.

[0042] Preferably, the pulsed electric field is a unipolar or bipolar high-voltage electric pulse with a pulse width of 10-100ns, a frequency of 0.5-10MHz, and an amplitude of 0-±5kV.

[0043] Preferably, the pulsed electric field is a pulsed electric field with a pulse width of 10-100ns, a frequency of 0.5-10MHz, and a value of unipolar 0-5kV or bipolar 0-±5kV.

[0044] Preferably, the products include, but are not limited to, medical devices, pharmaceutical compositions, and fields.

[0045] Preferably, the system is used in the preparation of an apparatus for killing glioma cells, targeting and inhibiting glioma activity, inhibiting glioma cell division and migration, inhibiting tight junction protein ZO-1 expression and / or reducing TEER; or the pulsed electric field is used in the preparation of a field and / or pharmaceutical composition for killing glioma cells, targeting and inhibiting glioma activity, inhibiting glioma cell division and migration, inhibiting tight junction protein ZO-1 expression and / or reducing TEER.

[0046] The beneficial effects of this invention are as follows:

[0047] 1. This invention utilizes an ultra-high frequency extremely short pulse electric field glioma ablation system. By integrating ultra-high frequency (>1MHz) extremely short pulse (nanosecond level) ablation with dynamic impedance matching control, it overcomes the technical difficulty of simultaneously achieving ablation treatment and opening the blood-brain barrier in existing technologies, and is expected to achieve more precise and safer brain tumor treatment.

[0048] 2. This invention directly ablates glioma tissue using an ultra-high frequency, extremely short pulse electric field while selectively opening the blood-brain barrier. This can assist in the delivery of chemotherapy / targeted drugs, enabling non-thermal ablation of deep tumors that are difficult to operate on (such as brainstem gliomas). Simultaneously, it enhances the permeability of immunotherapy drugs or nanomedicines, reduces the side effects of traditional radiotherapy / chemotherapy, and achieves an integrated "ablation + drug synergistic treatment" solution.

[0049] 3. In terms of technical performance, this invention employs a unique combination of 0.5-10MHz high frequency and 10-100ns ultra-short pulses. Its high-frequency characteristics effectively suppress the interference of tissue heterogeneity (such as regions with different conductivity and dielectric constants) on the electric field distribution, improving the uniformity of the electric field in the ablation area and simultaneously achieving uniform control of the blood-brain barrier opening depth. The nanosecond-level pulse width combined with high-frequency oscillation characteristics creates a local high-gradient field at the cell membrane level, achieving selective destruction of subcellular structures (ablation boundary accuracy up to ±0.3mm); on the other hand, it forms precise transmembrane potential regulation at the blood-brain barrier endothelial cell level, achieving selective and reversible opening of tight intercellular connections (opening boundary control accuracy up to ±0.5mm). The ultra-short pulse duration (10-100ns) is far shorter than the tissue thermal relaxation cycle (microsecond level), and combined with the rapid turn-off capability of the solid-state switch (turn-off time <5ns), it completely avoids Joule heat accumulation (measured temperature rise <0.5℃), significantly improving treatment safety. Through multimodal image-guided navigation and real-time impedance feedback, electrode layout and pulse parameters (frequency / pulse width / polarity) are dynamically optimized, precisely compensating for changes in local tissue conductivity and ensuring that the treatment area is highly consistent with the preset target area. The unique bipolar pulse mode further reduces the risk of neuronal electrical stimulation, achieving completely painless treatment.

[0050] 4. This invention successfully overcomes three major technical challenges in the treatment of gliomas using existing high-frequency pulsed electric field technology. First, through the synergistic design of ultra-high frequency (>1MHz) and extremely short pulse width (10-100ns), the contradiction between high cell selectivity and low heat accumulation in traditional microsecond pulsed electric field spectral characteristics is resolved, increasing the effective high-frequency component ratio to over 60% while maintaining subcellular-scale selectivity and controlling heat accumulation to within 0.5℃. Second, by employing dynamic impedance matching and intelligent electrode array technology, the adaptability problem of fixed electrode structures in complex brain tissue environments is overcome, increasing energy utilization from less than 40% to over 75%. Finally, it innovatively integrates ablation therapy and blood-brain barrier regulation functions onto a single platform, achieving precise spatiotemporal coordination through dual-frequency pulse coupling technology, overcoming the technical bottleneck of traditional methods' inability to simultaneously complete tumor removal and enhanced drug delivery. These breakthroughs make this system significantly superior to existing technologies in terms of treatment depth, precision, and safety, providing a completely new solution for the treatment of gliomas. Attached Figure Description

[0051] Figure 1 A diagram showing the components of an ultra-high frequency, extremely short pulse electric field glioma ablation system;

[0052] Figure 2 A schematic diagram of minimally invasive electrode ablation of glioma;

[0053] Figure 3 This is a structural diagram of the ultra-high frequency extremely short pulse electric field glioma ablation system according to an embodiment of the present invention;

[0054] Figure 4 This is another structural diagram of the ultra-high frequency extremely short pulse electric field glioma ablation system according to an embodiment of the present invention;

[0055] Figure 5 This is a structural diagram of the pulse generation module according to an embodiment of the present invention;

[0056] Figure 6 This is a schematic diagram illustrating the pathway by which ultra-high frequency, extremely short pulse electric fields kill tumor cells at multiple levels and activate the immune response.

[0057] Figure 7 The figure shows the experimental results of the killing effect of different parameters of ultra-high frequency extremely short pulse electric field on tumor cells. Figure 7 -A is a graph showing the killing effect on tumor cells under the parameters of field strength 50kV / cm, unipolarity, frequency within the pulse train 0.5MHz, and single pulse width 10-100ns; Figure 7 -B is a graph showing the killing effect on tumor cells under the parameters of field strength 50kV / cm, unipolarity, single pulse width 50ns, and pulse train frequency 0.5-10MHz; Figure 7-C is a graph showing the killing effect on tumor cells under the following parameters: field strength 50kV / cm, single pulse width 50ns, pulse train frequency 2MHz, and single and bipolar parameters. Figure 7 -D is a graph showing the killing effect on tumor cells under the parameters of a single pulse width of 50ns, a pulse train frequency of 2MHz, unipolarity, and a field strength of 10-50kV / cm.

[0058] Figure 8 The figure shows the experimental results of the windowing effect in glioma cells under an ultra-high frequency, extremely short pulse electric field.

[0059] Figure 9 The figure shows the experimental results related to changes in the cytoskeleton of glioma cells before and after ultra-high frequency pulsed electric fields. Figure 9 -A is a diagram showing the cytoskeleton morphology of mouse glioma cells GL261 before treatment with an ultra-high frequency pulsed electric field under an atomic force microscope. Figure 9 -B is a diagram of the cytoskeleton structure of the human glioma cells U251 that were treated. Figure 9 -C is a magnified view of a local area of ​​U251 cells, showing a clear arrangement of filamentous skeletons; Figure 9 -D is a diagram showing the cytoskeleton morphology of mouse glioma cells GL261 after treatment with an ultra-high frequency pulsed electric field under an atomic force microscope. Figure 9 -E shows the changes in the skeletal structure of U251 cells after treatment; Figure 9 -F is a magnified image of U251 cells, showing that their filamentous skeleton has depolymerized and formed clustered structures; Figure 9 -G is an immunofluorescence staining image of human glioma cells U251 before treatment with ultra-high frequency pulsed electric field. Orange represents actin F-actin, green represents tubulin, and blue represents the cell nucleus. The cytoskeleton, mainly composed of actin and tubulin, is arranged in a dense filamentous pattern to maintain cell morphology. Figure 9 -H is an immunofluorescence image of the cytoskeleton of U251 cells after treatment with an ultra-high frequency pulsed electric field. Tubulin depolymerizes into clumps, actin depolymerizes and accumulates in the cell membrane, and the overall cell morphology shrinks.

[0060] Figure 10 The figure shows the experimental results of DNA damage in glioma cells induced by ultra-high frequency extremely short pulse electric fields. Figure 10 -A shows the immunofluorescence of human glioma cells before and after treatment with an ultra-high frequency pulsed electric field. In the figure, green represents gamma.H2AX protein, red represents 53BP1 protein, both of which are DNA damage marker proteins, and blue represents the cell nucleus. Overall, the higher the dose of the ultra-high frequency extremely short pulsed electric field, the greater the proportion of cells with positive DNA damage. Figure 10-B is an immunoblot image of human glioma cells U251 before and after pulse treatment. It can be seen that after pulse treatment, the amount of gamma.H2AX protein increased, and PARP1 protein was cleaved into cleaved-PARP1, activating the PARP1-related apoptosis pathway. Figure 10 -C is the cell cycle distribution of residual cells 24 hours after treatment with ultra-high frequency ultra-short pulse electric field. The proportion of S phase cells increased significantly after treatment, indicating that DNA damage induced G2 / M phase arrest and slowed down cell division and proliferation.

[0061] Figure 11 The figure shows the experimental results related to the increase of intracellular dsDNA after treatment with an ultra-high frequency extremely short pulse electric field. In the figure, red represents double-stranded DNA fragments (dsDNA), green represents cyclic guanosine monophosphate-adenosine synthase (cGas) protein, and blue represents the cell nucleus. Before and after treatment with an ultra-high frequency extremely short pulse electric field, the nuclei of human glioblastoma cells U87-MG shrank. After DNA damage, DNA fragments were released into the cytoplasm, activating cGas protein and thus inducing immunogenic cell death.

[0062] Figure 12 This is an in vivo imaging image of a mouse with an ultra-high frequency, extremely short pulse electric field inhibiting the development of orthotopic glioma tumors. Figure 12 -A~ Figure 12 -C is an in vivo imaging image of three control group mice; Figure 12 -D~ Figure 12 -F represents in vivo imaging of the three experimental groups of mice. After confirming the tumor formation effect on the 9th day after in situ inoculation of glioma, ultra-high frequency extremely short pulse electric field ablation was performed. Subsequently, the tumor ablation effect was confirmed by in vivo imaging on the 2nd and 7th days after surgery.

[0063] Figure 13 This is a diagram showing the experimental results of an in vitro blood-brain barrier model. Figure 13 -A is a graph showing the measurement of TEER resistance at various time points during the construction of the transwell-based in vitro blood-brain barrier model to characterize the degree of blood-brain barrier model construction. When the TEER resistance gradually increases and tends to stabilize, it indicates that the blood-brain barrier model has been successfully constructed. After treatment with an ultra-high frequency extremely short pulse electric field, the TEER resistance shows a significant decrease, indicating that the surface blood-brain barrier has been damaged. Figure 13 -B shows the results of immunofluorescence staining of tight junction protein ZO-1 in an in vitro blood-brain barrier model after fixation with 4% paraformaldehyde. In the pulsed area, the blue cell nuclei are preserved while the green tight junction protein disappears, indicating that the tight junction structure between cells in the in vitro blood-brain barrier model was damaged after treatment with an ultra-high frequency ultra-short pulse electric field.

[0064] Figure 14 The figure shows the experimental results related to the permeability of the blood-brain barrier in mice induced by ultrashort pulse electric fields. Figure 14-A is the Evans indigo staining diagram of the control group. Figure 14 -B is the Evans indigo staining diagram of the experimental group;

[0065] Figure 15 The graph shows the change in Evans blue concentration in the mouse brain over time. Figure 15 -A is a standard curve plotted using the absorbance of different concentrations of Evans blue dye at a wavelength of 625 nm. Figure 15 -B is a graph showing the absorption of the supernatant at 625 nm after stimulating the mouse brain with ultra-high frequency and extremely short pulses, followed by cardiac perfusion at different time points. The whole brain tissue was then removed and placed in formamide, crushed, and placed at 55°C for 24 hours. The supernatant was then centrifuged and the absorbance of the supernatant was measured. The results show that the blood-brain barrier of the mouse was momentarily permeable after pulse treatment and returned to normal level after 24 hours.

[0066] In the above figures, 100 is the data acquisition and analysis module, 200 is the control module, 300 is the pulse generation module, 301 is the pulse generator, 302 is the pulse output switch, 400 is the ablation module, 401 is the minimally invasive single needle electrode, 402 is the flat grounding electrode, and 500 is the wire. Detailed Implementation

[0067] The technical solution of the present invention will be described more clearly and completely below with reference to specific embodiments. Obviously, the described embodiments are only a part of the embodiments of the present invention, and not all of them. Therefore, based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the protection scope of the present invention.

[0068] In the description of this invention, unless otherwise stated, "a plurality of" means two or more; the terms "upper," "lower," "left," "right," "inner," "outer," "front end," "rear end," "head," "tail," etc., indicate the orientation or positional relationship based on the orientation or positional relationship shown in the accompanying drawings, and are only for the convenience of describing the invention and simplifying the description, and do not indicate or imply that the system, device, or element referred to must have a specific orientation, or be constructed and operated in a specific orientation, and therefore should not be construed as a limitation of the invention. Furthermore, the terms "first," "second," "third," etc., are used for descriptive purposes only and should not be construed as indicating or implying relative importance.

[0069] In the description of this invention, it should be noted that, unless otherwise explicitly specified and limited, the terms "connected" and "linked" should be interpreted broadly. For example, they can refer to a fixed connection, a detachable connection, or an integral connection; they can refer to a mechanical connection or an electrical connection; they can refer to a direct connection or an indirect connection through an intermediate medium. Those skilled in the art can understand the specific meaning of the above terms in this invention according to the specific circumstances.

[0070] The system and principles of the present invention will now be further explained with reference to the accompanying drawings.

[0071] In some embodiments, such as Figure 3 and Figure 4 As shown, the system for ablating tumor tissue based on ultra-high frequency extremely short pulse electric field of the present invention includes: a data acquisition and analysis module 100, a control module 200, a pulse generation module 300, and an ablation module 400; wherein, the data acquisition and analysis module 100 is used to acquire and analyze MRI, CT, and ultrasound detection results to obtain the location and size of the ablation target area and the electrode implantation location; the control module 200 is connected to the data acquisition and analysis module 100 to generate treatment pulse parameters based on the image detection results of the data acquisition and analysis module 100; the control module 200 is connected to the pulse generation module 300 to control the pulse generation module 300 to output pulses with set parameters or cut off the pulse electric field; the ablation module 400 is provided with a minimally invasive single needle electrode 401 and a flat plate grounding electrode 402; the pulse generation module 300 is connected to the minimally invasive single needle electrode 401 and the flat plate grounding electrode 402 respectively through wires 500; the pulse generation module 300 outputs a pulse electric field and targets and ablates the tumor tissue through the ablation module 400.

[0072] In some embodiments, the pulse generation module is used to output a unipolar or bipolar high-voltage pulse with a pulse width of 10-100ns, a frequency of 0.5-10MHz, and an amplitude of 0-±5kV.

[0073] In some embodiments, the pulse generation module is used to output a pulsed electric field with a pulse width of 10-100ns, a frequency of 0.5-10MHz, and a value of unipolar 0-5kV or bipolar 0-±5kV.

[0074] In some embodiments, such as Figure 5 As shown, the pulse generation module 300 includes a pulse generator 301 and a pulse output switch 302; the pulse output switch 302 is connected between the pulse generator 301 and the ablation module 400 to control the on / off state of the pulse electric field.

[0075] In some embodiments, the minimally invasive single-needle electrode 401 is implanted in the center of the tumor tissue, and the flat ground electrode 402 is attached to the skin surface to form a closed circuit.

[0076] For example, the data acquisition and analysis module 100 can select a multimodal image navigation system. Its function is to collect and analyze MRI, CT, and other image detection results through import and other methods, and then output the location and size of the ablation target area and the electrode implantation location. Preferably, the ablation target area and the open area of ​​the blood-brain barrier can be accurately delineated using multimodal image fusion technology (MRI / CT), and a geometric model of the target tissue can be constructed using a three-dimensional reconstruction algorithm. Electric field simulation calculations are performed based on the conductivity distribution characteristics of brain tissue to optimize the spatial layout of the single-needle electrode and the flat electrode, ensuring that the electric field coverage is highly matched to the lesion geometry and treatment requirements. Under real-time image navigation guidance, the minimally invasive single-needle electrode is precisely implanted into the core position of the target area, achieving sub-millimeter-level positioning accuracy.

[0077] For example, the control module 200 can select a human-computer interaction control terminal. Based on the image detection results from the data acquisition and analysis module 100, it generates treatment pulse parameters; wherein the control parameters can be specifically set pulse width, frequency, amplitude, and polarity, and the set pulse width, frequency, amplitude, and polarity can be specific signal values ​​selected within the range of 10-100ns, 0.5-10MHz, 1-20kV, 0-5kV unipolar, or 0-±5kV bipolar. It should be noted that in actual implementation, different values ​​will be selected depending on the symptoms of the treatment subject; this embodiment does not limit the specific value selection of the control parameters.

[0078] For example, the pulse generator 301 may be a solid-state pulse generator.

[0079] The system employs an intelligent control terminal to dynamically optimize pulse parameter combinations, including pulse widths of 10-100 ns, frequencies of 0.5-10 MHz, electric field strengths of 5-50 kV / cm (pulse amplitude 1-5 kV), and adjustable polarity modes. A solid-state pulse generator produces ultra-high frequency, extremely short pulse width square wave pulse sequences. The pulse energy, transmitted through a needle-plate electrode pair, forms a three-dimensional focused electric field, simultaneously achieving dual therapeutic effects on a nanosecond timescale: firstly, it induces hyperpolarization of the target cell membrane, triggering multiple biological effects such as calcium ion influx, mitochondrial membrane potential collapse, and chromatin fragmentation, leading to non-thermal apoptosis; secondly, it reversibly regulates the connections between endothelial cells and pericytes in the blood-brain barrier, enhancing drug permeability. Throughout the process, the extracellular matrix and vascular and neural structures are completely preserved, minimizing damage to surrounding normal tissues.

[0080] Preferably, the electric field distribution is monitored in real time during the procedure using multimodal imaging, and pulse parameters are dynamically adjusted using impedance feedback to effectively compensate for electric field distortions caused by tissue heterogeneity, ensuring that the ablation boundary and the blood-brain barrier opening range are controlled with sub-millimeter precision. After treatment, the system automatically generates an electric field energy deposition map, a tissue damage quantitative assessment report, and a blood-brain barrier opening effect assessment report, providing clinicians with objective evidence to verify the treatment effect. This system innovatively integrates tumor ablation and blood-brain barrier regulation functions into a single platform, achieving synergistic treatment through precise spatiotemporal control, providing a novel solution for the treatment of central nervous system diseases such as gliomas.

[0081] Example 1. Ultra-high frequency extremely short pulse electric field glioma ablation system

[0082] A high-frequency, extremely short pulse electric field ablation system for gliomas, comprising the following components:

[0083] 1) Ultra-high frequency extremely short pulse generator, using solid-state switching technology, can output ultra-high frequency high voltage pulses at the nanosecond level (10-100ns), with values ​​of unipolar 0-5kV or bipolar 0-±5kV and frequencies of 0.5MHz-10 MHz.

[0084] 2) Minimally invasive single-needle electrode and flat grounded electrode: The single-needle electrode is inserted into the tumor area through minimally invasive puncture, and the flat grounded copper foil electrode is attached to the patient's body surface (similar to forming a needle plate discharge for targeted ablation of tissue), thereby forming a three-dimensional electric field coverage of the targeted ablation area in the patient's body.

[0085] 3) A multimodal image navigation system can utilize imaging techniques such as MRI / CT to measure parameters such as the location and size of the patient's ablation target area. Combined with electric field simulation, it analyzes the placement of the minimally invasive single-needle electrode and the flat grounded electrode, as well as the applied ultra-high frequency extremely short pulse parameters, thereby simulating and determining the tissue ablation range and formulating an ablation plan. Furthermore, it can integrate ultrasound / MRI images in real-time during the procedure to guide the electrode needle placement and assess the ablation efficacy in real-time, ensuring precise electrode positioning and control of the ablation boundary.

[0086] 4) Human-computer interaction control terminal with an integrated operating interface, allowing for the setting of treatment pulse parameters and evaluation of ablation efficacy. It supports operators in customizing treatment protocols and provides real-time visualization of the ablation process.

[0087] The components and their collaborative workflow of the ultra-high frequency, ultra-short pulse electric field minimally invasive glioma treatment system are as follows: Figure 1As shown, the system comprises four key components: an ultra-high frequency ultra-short pulse generator, minimally invasive electrodes, a multimodal image navigation system, and a human-computer interaction control terminal. These components work together through precise connections to achieve accurate treatment of gliomas. After a patient is diagnosed with a glioma, they first undergo imaging examinations, including high-resolution MRI, functional imaging, and contrast-enhanced scans, to obtain crucial information such as the precise location, volume, and spatial relationship of the intracranial tumor to functional areas. The preoperative imaging data is imported into the multimodal image navigation system to reconstruct a three-dimensional model of the patient's skull, and the tumor lesion is precisely marked and outlined. Next, in a computer-aided modeling environment, the optimal electrode implantation path is planned based on the spatial distribution and morphology of the tumor, and electric field simulation analysis is used to simulate the distribution of the electric field in the tumor area during treatment. The system automatically determines whether an effective treatment intensity can be achieved within the tumor volume and constrains the electric field intensity in non-target tissue areas to prevent irreversible damage to neurological functional areas. After confirming the electrode path is correct, the surgeon performs the electrode implantation operation using a stereotactic device, ensuring that the electrode tip accurately reaches the planned treatment area. Subsequently, simulation parameters are retrieved from the human-computer interaction control terminal and sent to the ultra-high frequency ultra-short pulse generator. The terminal supports real-time adjustment and monitoring of parameters such as pulse waveform, amplitude (0-±5kV), frequency (0.5-10MHz), pulse width (10-100ns), and number of treatment pulses. Finally, under the closed-loop control architecture, the system controls the generator to output the set electrical pulses, which are released to the tumor area through the implanted minimally invasive electrodes, completing the precise ablation of the glioma.

[0088] Example 2. Ablation of glioma using ultra-high frequency extremely short pulse electric field

[0089] A schematic diagram of minimally invasive electrodes used in glioma ablation is shown below. Figure 2 As shown in the image, during treatment, a stereotactic single-needle electrode (positive electrode) is percutaneously implanted into the center of the lesion to achieve precise electric field coverage of the tumor area. In the image, this electrode enters the cranium through the scalp, with its tip directly facing the tumor core to ensure that the electric field intensity reaches the treatment threshold within the tumor volume. Simultaneously, a flexible sheet-like negative electrode is attached to the skin surface of the patient's chin, utilizing its excellent fit and conductivity to construct a closed circuit, minimizing the impact of the current path on normal brain tissue. The electrodes are connected to an external pulse generator via wires, and the treatment parameters and output timing are controlled by a human-machine interface terminal.

[0090] Example 3. Multi-level killing of tumor cells and activation of immune response pathways by ultra-high frequency extremely short pulse electric fields.

[0091] This invention explores the mechanism by which ultra-high frequency, extremely short pulse electric fields kill tumor cells and activate immune response pathways. For example... Figure 6As shown, ultra-high frequency, ultra-short pulsed electric field treatment can induce multilayer cell death and activate immune response pathways. First, the electric field can disrupt cytoskeleton structures, such as microtubules and microfilaments, leading to changes in cell morphology and dysfunction. Second, the electric field induces DNA damage, causing DNA strand breaks and genomic instability, activating cell repair mechanisms; if repair fails, apoptosis or necrosis is induced. Furthermore, the ultra-high frequency, ultra-short pulsed electric field can also activate the cGAS-STING immune pathway, activating downstream type I interferon responses by recognizing intracellular and extracellular DNA, enhancing antigen presentation and immune cell activation. The antigens and danger signals released during cell death can further promote T cell responses and immune system activation, showing potential, particularly in tumor immunotherapy.

[0092] Example 4. Verification of the efficacy of the ultra-high frequency, extremely short pulse electric field glioma ablation system.

[0093] (1) Effects of different parameters of ultra-high frequency ultra-short pulse electric field on tumor cell killing effect

[0094] One hour after the pulsed electric field, tumor cell activity was assessed using a CCK-8 cell proliferation assay. The following parameters were tested: ① Electric field strength 50 kV / cm (i.e., pulse amplitude 5 kV, electrode spacing 1 mm), unipolarity, pulse train frequency 0.5 MHz, single pulse width 10-100 ns; ② Electric field strength 50 kV / cm (i.e., pulse amplitude 5 kV, electrode spacing 1 mm), unipolarity, single pulse width 50 ns, pulse train frequency 0.5-10 MHz; ③ Electric field strength 50 kV / cm (i.e., pulse amplitude 5 kV, electrode spacing 1 mm), single pulse width 50 ns, pulse train frequency 2 MHz, unipolar / bipolarity; ④ Single pulse width 50 ns, pulse train frequency 2 MHz, unipolarity, electric field strength 10-50 kV / cm (i.e., pulse amplitude 1-5 kV, electrode spacing 1 mm). Comparison of tumor cell activity under different pulse parameter configurations is shown below. Figure 7 As shown.

[0095] (2) Windowing effect of glioma cells under ultra-high frequency ultra-short pulse electric field

[0096] Human glioma cells U-87MG and human astrocytes SVG p12 were treated with an ultra-high frequency, ultra-short pulsed electric field for 1 hour, and cell viability was assessed using a CCK-8 cell proliferation assay. The window effect between glioma cells and normal cells after ultra-high frequency pulsed electric field treatment was as follows: Figure 8 As shown, under specific pulsed electric field parameter configurations (in this embodiment, electric field strength 50kV / cm, single pulse width 50ns, frequency within the pulse train 2MHz, bipolarity, 20 trains), targeted inhibition of glioma activity can be achieved without significantly inhibiting normal cell physiological functions.

[0097] (3) Ultra-high frequency extremely short pulse electric fields disrupt the cytoskeleton

[0098] After applying a unipolar pulsed electric field with a strength of 30 kV / cm (i.e., pulse amplitude of 3 kV, electrode spacing of 1 mm), a pulse width of 50 ns, and a frequency of 2 MHz using needle electrodes, human glioma cell samples (LN229) were immediately fixed with 4% paraformaldehyde to maintain cytoskeleton morphology. After cell membrane removal, analysis was performed using atomic force microscopy and immunofluorescence staining. Figure 9 As shown, before pulse treatment, the tubulin filaments are densely distributed to support the cell's geometric morphology system and maintain normal cellular physiology and communication functions; after pulse treatment, the tubulin fragments, depolymerizes, and shrinks towards the cell nucleus, indicating that ultra-high frequency ultra-short pulse electric fields can effectively inhibit the division and migration abilities of tumor cells, leading to immunogenic death of tumor cells, releasing tumor antigens through scaffold damage, and activating immune responses (such as calreticulin exposure).

[0099] (4) Ultra-high frequency extremely short pulse electric field induces DNA damage in glioma cells.

[0100] Human glioma cells U251 were fixed with 4% paraformaldehyde or had their proteins extracted via RIPA after 1 hour of exposure to a unipolar pulsed electric field with a field strength of 20 kV / cm (i.e., pulse amplitude of 2 kV, electrode spacing of 1 mm), pulse width of 50 ns, and frequency of 2 MHz. Detection was performed using immunofluorescence and Western blotting. Results are as follows: Figure 10 As shown, the expression of Gamma.H2AX and 53BP1 proteins is upregulated, PARP1 protein is cleaved, the DNA double-strand break repair pathway is activated, DNA replication in the S phase of the cell cycle is arrested, and the apoptosis program is initiated.

[0101] (5) Increased intracellular dsDNA after treatment with ultra-high frequency extremely short pulse electric field

[0102] Human glioma cells LN229 were treated for 3 hours with a bipolar pulsed electric field of 20 kV / cm (i.e., pulse amplitude 2 kV, electrode spacing 1 mm), pulse width 50 ns, and frequency 2 MHz. Cell samples were then fixed with 4% paraformaldehyde and detected by immunofluorescence. Results are as follows: Figure 11 As shown, after the nuclear membrane is ruptured by an ultra-high frequency, extremely short pulse electric field, fragmented double-stranded DNA fragments diffuse into the cytoplasm. Abnormal double-stranded DNA (dsDNA) can be recognized by cGAS, activating its catalytic function to synthesize the secondary signaling molecule cGAMP, which in turn activates the STING pathway, induces the expression of type I interferon and inflammatory factors, and initiates the innate immune response, playing an important role in antiviral, antitumor, and immunogenic cell death.

[0103] (6) Ultra-high frequency extremely short pulse electric field inhibits the development of orthotopic glioma in mice.

[0104] The method for constructing an orthotopic mouse model of glioma includes: using 1×10 7 / mL GL261-luci cell suspension was administered to C57BL / 6 mice. After intraperitoneal anesthesia with 0.75% sodium pentobarbital at 50 mg / kg body weight, the hair at the puncture site was scraped and disinfected. The mouse skull was fixed using a stereotactic apparatus, and a longitudinal incision was made in the scalp to locate the anterior fontanelle. The puncture point was selected 1 mm posterior to the anterior fontanelle and 2 mm lateral to the right of the midline. The needle was inserted perpendicularly for 3 mm, and GL261-luci cell suspension was injected via a microinfusion pump at a rate of 2.5 μl / min. The needle was retained for 2 minutes and then withdrawn. The puncture site was sealed with bone wax, and the scalp incision was sutured. Nine days later, after confirming tumor formation through live imaging, a bipolar pulsed electric field of 3 kV, 50 ns pulse width, and 2 MHz frequency was applied. The live imaging results are as follows: Figure 12 As shown, gliomas in mice were significantly inhibited or even almost disappeared one week after pulse treatment.

[0105] (7) In vitro blood-brain barrier model experiment

[0106] An in vitro blood-brain barrier model was constructed using the Transwell system. 200 μL of the mouse brain microvascular endothelial cell line bEND3 was injected at a concentration of 1.5 × 10⁻⁶. 5 cells / cm 2 The trans-endothelial resistance (TEER) model was inoculated at a density of 0.4 μm polycarbonate membrane into the upper chamber of a Transwell insert. Transendothelial resistance was monitored every 48 hours, and the culture medium was replaced with fresh medium. After 8 days, the TEER resistance stabilized. A bipolar high-frequency ultrashort pulse electric field with a pulse width of 50 ns, an amplitude of 3 kV, an intra-string frequency of 2 MHz, 100 intra-string pulses, and 100 pulse trains was then applied to the Transwell in vitro blood-brain barrier model. Immunofluorescence staining and quantitative analysis were performed on the model one hour later. Results are as follows: Figure 13 As shown, the expression level of the tight junction protein ZO-1 in the pulsed region was significantly lower than that in the control group. This result suggests that high-frequency, extremely short pulsed electric fields lead to a decrease in TEER by disrupting the tight junctions between brain microvascular endothelial cells, thus improving the permeability of the in vitro blood-brain barrier model.

[0107] (8) Ultrashort pulse electric field induces blood-brain barrier permeability in mice

[0108] Using an ultrashort pulse electric field therapy system, a bipolar ultrashort pulse electric field (50 ns pulse width, 3 kV amplitude, 2 MHz intra-sequence frequency, 100 intra-sequence pulses, 100 pulse trains) was applied to the brain of 8-week-old mice. Immediately afterward, 2% (w / v) Evans blue dye was injected via the tail vein at a volume of 3 ml / kg based on the mouse's body weight. One hour later, the mice were re-anesthetized, and their hearts were perfused with physiological saline. After the fluid flowing from the right atrium became clear, the mouse brains were removed to observe the distribution of Evans blue dye in the brain. The results are as follows: Figure 14 As shown, Evans blue permeates across the blood-brain barrier in healthy mice after an ultrashort pulse electric field is applied, and the distribution of Evans blue dye in the brains of mice treated with the ultrashort pulse electric field is wider, indicating that the permeability of the blood-brain barrier is significantly improved.

[0109] (9) Changes in Evans blue concentration in mouse brain over time

[0110] Evans blue dye was injected into mice at different time points to assess blood-brain barrier permeability. First, Evans blue dye was serially diluted with formamide, and absorbance at 625 nm was measured using an ELISA reader to plot a standard curve. After applying an ultrashort pulse electric field with a pulse width of 50 ns, an amplitude of 3 kV, an intra-string frequency of 2 MHz, 100 intra-string pulses, and 100 pulse trains to the mouse brain, Evans blue dye was injected into the tail vein at different time points. Following cardiac perfusion, the whole brain was harvested, ground in 650 μL of formamide, heated at 55°C for 24 hours, and centrifuged at 12000g for 20 minutes. The supernatant was then measured for absorbance at 625 nm. Results are as follows: Figure 15 As shown, the blood-brain barrier in mice can be temporarily opened after treatment with an ultrashort pulse electric field and gradually recovers within 24 hours.

[0111] Example 5

[0112] Replacing the original metal electrodes with PEDOT:PSS conductive polymer patches reduced the surface impedance from 50Ω to 15Ω (1kHz test). After 100 bending cycles, the conductivity decreased by less than 5%, significantly improving wearing comfort, sports adaptability, and fit stability while maintaining conductivity.

[0113] Example 6

[0114] The output parameters were adjusted to: frequency 1MHz, field strength 15kV / cm (i.e., pulse amplitude 1.5kV). Animal experiments verified that: ① it could still effectively induce electroporation of tumor tissue and activation of the cGAS pathway; ② it was compatible with low-cost generators; ③ combined with pulse number compensation design, i.e., by calculating the same dose, the electric field strength could be appropriately reduced to increase the number of pulse trains, which could reduce the risk of dielectric breakdown. Live cell imaging showed that the cell death rate decreased from 6% to 1% (p<0.01).

[0115] Example 7

[0116] Optimized pulse sequence: ① Reducing the number of pulses from 100 to 50 shortened the blood-brain barrier opening time by 38% (p<0.05); ② The 30-pulse regimen maintained significant tumor-suppressive activity in vivo, with a tumor inhibition rate >85%. Both regimens maintained a 3kDa dextran permeability 2.1±0.3 times that of the control group, and increased doxorubicin transport efficiency by 2.7 times (HPLC verification).

[0117] Example 8

[0118] The user interface has been optimized to retain only core functions such as "treatment mode," "electrode confirmation," and "pulse activation," reducing the intraoperative error rate by 72%. Stress testing has confirmed improved system stability, while the design of hiding redundant parameters shortens new user training time by 40%.

Claims

1. A system for ablating tumor tissue based on ultra-high frequency extremely short pulse electric field, characterized in that, The system includes: Data acquisition and analysis module: used to acquire and analyze MRI, CT, and ultrasound test results to obtain the location and size of the ablation target area and the electrode implantation location; Control module: The control module is connected to the data acquisition and analysis module to generate treatment pulse parameters based on the image detection results of the data acquisition and analysis module; Pulse generating module: The control module is connected to the pulse generating module to control the pulse generating module to output pulses with set parameters or to cut off the pulse electric field; Ablation module: It is equipped with a minimally invasive single-needle electrode and a flat grounded electrode; the pulse generation module is connected to the minimally invasive single-needle electrode and the flat grounded electrode respectively through wires; the pulse generation module outputs a pulsed electric field and targets and ablates tumor tissue through the ablation module.

2. The system according to claim 1, characterized in that, The pulse parameters include pulse amplitude, polarity, pulse width, and frequency.

3. The system according to claim 1, characterized in that, The pulse generation module is used to output unipolar or bipolar high-voltage pulses with a pulse width of 10-100ns, a frequency of 0.5-10MHz, and an amplitude of 0-±5kV.

4. The system according to claim 3, characterized in that, The pulse generation module is used to output a pulsed electric field with a pulse width of 10-100ns, a frequency of 0.5-10MHz, and a value of unipolar 0-5kV or bipolar 0-±5kV.

5. The system according to claim 1, characterized in that, The pulse generation module includes a pulse generator and a pulse output switch; the pulse output switch is connected between the pulse generator and the ablation module to control the on / off state of the pulse electric field.

6. The system according to claim 1, characterized in that, The minimally invasive single-needle electrode is implanted in the center of the tumor tissue, and the flat ground electrode is attached to the skin surface to form a closed circuit.

7. The system according to claim 1, characterized in that, The data acquisition and analysis module is a multimodal image navigation system.

8. The system according to claim 1, characterized in that, The control module is a human-computer interaction control terminal.

9. The use of the system and / or pulsed electric field according to any one of claims 1-8 in the preparation of products for ablating glioma tissue, increasing blood-brain barrier permeability and / or inhibiting glioma development.

10. The use of the system according to any one of claims 1-8 in the preparation of products for killing glioma cells, targeting and inhibiting glioma activity, inhibiting glioma cell division and migration, inhibiting tight junction protein ZO-1 expression and / or reducing TEER.

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