Ultrasound methods and devices for enhanced cell collection in cytopathology

Abstract

A method of producing a cell sample of cytopathology, the method comprising ultrasonically irradiating a target tissue with ultrasonic energy having a focused beam that produces a shear wave within the tissue. A method of producing a cell sample of cytopathology, the method comprising ultrasonically irradiating a target tissue with a first intensity level of ultrasonic energy during a first time period, and ultrasonically irradiating the target tissue with a second intensity level of ultrasonic energy during a second time period, where the first intensity level is higher than the second intensity level. Related apparatus and methods are also described.

Description

[0001] Related applications

[0002] This application claims the benefit of priority to U.S. Provisional Patent Application No. 63 / 463,344, filed May 2, 2023, the contents of which are incorporated herein by reference in their entirety. Technical Field

[0003] This disclosure relates, in some embodiments thereof, to medical systems and methods for separating target epithelium, and particularly to ultrasound methods for separating target epithelium, and to ultrasound devices constructed and / or programmed to use such methods, and to programming ultrasound devices to use such methods. Background Technology

[0004] Pancreatic cancer is an extremely deadly cancer, with its high mortality rate largely attributed to its tendency to be detected at an advanced stage. The lack of reliable and / or non-invasive tests for early detection of pancreatic cancer is a major contributing factor. Attempts have been made to harvest pancreatic cells from pancreatic juice for early detection, but the sensitivity of this test is low due to the scarcity of naturally occurring pancreatic cells in pancreatic juice.

[0005] Additional background technology includes:

[0006] U.S. Patent Application No. 17 / 007,953, filed on August 31, 2020, entitled "Device for Inducing Exfoliation of Cells and / or Tissue Fragments for Enhanced Cytopathologic Cell Collection."

[0007] The public information of all references mentioned above and throughout this specification, as well as the public information of all references mentioned in those references, is hereby incorporated into this document by reference. Summary of the Invention

[0008] This disclosure relates, in some embodiments thereof, to an ultrasound method for breaking up target epithelium using ultrasonic shear waves, to an ultrasound device constructed and / or programmed to use this method, and to programming an ultrasound device to use this method.

[0009] This disclosure relates, in some embodiments thereof, to an ultrasound method for enhancing the stripping of cells and / or molecules of a target epithelium, to constructing and / or programming an ultrasound device for using this method, and to programming an ultrasound device to use this method.

[0010] A broader aspect of some embodiments of the invention includes irradiating tissue with high-intensity focused ultrasound for a period of time to generate shear waves that partially sever the target epithelium by creating transverse breaks between cells, thereby inducing the shedding of cell and tissue debris.

[0011] A broad aspect of some embodiments of the invention includes irradiating tissue with high-intensity focused ultrasound during a first time period to generate shear waves that partially segment the target epithelium by creating transverse breaks between cells, followed by injection of a microbubble contrast agent, and irradiating the tissue with low-intensity ultrasound during a second time period (which elevates the partially segmented epithelium from below) to induce the shedding of cells and tissue debris.

[0012] According to one aspect of some embodiments of this disclosure, a method for inducing the shedding of cells and molecules from a target epithelium is provided, the method comprising irradiating the target tissue with a high-energy focused beam of ultrasound with a mechanical index (MI) greater than 0.4 to generate shear waves within the tissue.

[0013] According to one aspect of some embodiments of this disclosure, a method for generating cellular pathological samples is provided, the method comprising irradiating a target tissue with ultrasound energy having a focused beam, the focused beam generating shear waves within the tissue.

[0014] According to one aspect of some embodiments of this disclosure, a method for inducing the shedding of cells and molecules from a target epithelium is provided, the method comprising applying a focused ultrasound beam, wherein the ultrasound beam is arranged to: generate a mechanical index (MI) in the range of 0.4 to 1.8 to generate a focused beamwidth in the range of 2 mm to 10 mm / at the focal point; generate a pulse width in the range of 100 microseconds to 800 microseconds; and generate a pulse repetition rate in the range of 4 Hz to 20 Hz.

[0015] According to one aspect of some embodiments of this disclosure, a method for generating a cytopathological cell sample is provided, the method comprising irradiating a target tissue with ultrasound energy at a first intensity level during a first time period, and irradiating the target tissue with ultrasound energy at a second intensity level during a second time period, wherein the first intensity level is higher than the second intensity level.

[0016] According to some embodiments of this disclosure, the first strength level corresponds to a mechanical index (MI) in the range of 0.3 to 1.8.

[0017] According to some embodiments of this disclosure, during a first time period, the target tissue is irradiated with ultrasonic energy, including by ultrasonic irradiation with a focused ultrasonic beam.

[0018] According to some embodiments of this disclosure, the second intensity level corresponds to a mechanical index (MI) in the range of 0.1 to 0.4.

[0019] According to some embodiments of this disclosure, during the second time period, the target tissue is irradiated with ultrasonic energy, including by ultrasonic irradiation with a non-focused ultrasonic beam.

[0020] According to some embodiments of this disclosure, during a first time period, the target tissue is irradiated by ultrasonic energy, including by ultrasonic irradiation with ultrasonic pulses of repetitive pulse width in the range of 2 microseconds to 800 microseconds.

[0021] According to some embodiments of this disclosure, during the second time period, the target tissue is irradiated by ultrasonic energy, including by ultrasonic irradiation with ultrasonic pulses of repetitive duration in the range of 20 microseconds to 800 microseconds.

[0022] According to some embodiments of this disclosure, during a first time period, ultrasonic irradiation of the target tissue by ultrasonic energy includes repeating ultrasonic pulses at a rate ranging from 1 to 20 times per second.

[0023] According to some embodiments of this disclosure, during the second time period, the target tissue is irradiated with ultrasonic energy, including repeating ultrasonic pulses at a rate ranging from 1 to 100 times per second.

[0024] According to some embodiments of this disclosure, during a second time period, the target tissue is irradiated with ultrasound at a second intensity level, prior to the second time period, by providing an ultrasound contrast agent to the target tissue before irradiating the target tissue with ultrasound at a second intensity level during the second time period.

[0025] According to some embodiments of this disclosure, during the second time period, ultrasound energy is used to irradiate the target tissue, including providing an ultrasound contrast agent to the target tissue during the second time period.

[0026] According to some embodiments of this disclosure, the target tissue includes the pancreas.

[0027] According to some embodiments of this disclosure, the target tissue includes the prostate. According to several other embodiments of this disclosure, the target tissue includes the bladder.

[0028] According to some embodiments of this disclosure, it also includes: injecting a patient with a drug that induces pancreatic secretion; and collecting cytopathological samples by collecting pancreatic secretions.

[0029] According to some embodiments of this disclosure, the target tissue includes body locations selected from the group consisting of the mediastinum, pleura, pericardium, peritoneum, lungs, breast, salivary glands, meninges, pancreas, pancreatic duct, pancreatic cysts, kidneys, liver, prostate, bladder, and ovaries.

[0030] According to one aspect of some embodiments of this disclosure, a method for inducing exfoliation is provided, the method comprising ultrasonically irradiating a target tissue with ultrasonic energy in the range of 1.2 to 1.8.

[0031] According to some embodiments of this disclosure, ultrasonic irradiation of target tissue by ultrasonic energy includes ultrasonic irradiation by a focused ultrasonic beam.

[0032] According to some embodiments of this disclosure, ultrasonic irradiation of target tissue by ultrasonic energy includes ultrasonic irradiation by ultrasonic pulses with repetitive pulse widths in the range of 20 microseconds to 800 microseconds.

[0033] According to some embodiments of this disclosure, ultrasonic irradiation of target tissue by ultrasonic energy includes repeating ultrasonic pulses at a rate of 1 to 50 times per second.

[0034] According to one aspect of some embodiments of this disclosure, an ultrasonic energy source is provided that is programmed to perform ultrasonic irradiation using an ultrasonic irradiation procedure, the procedure comprising performing ultrasonic irradiation during a first time period using focused ultrasonic energy at a first intensity level designed to generate shear waves in the ultrasonically irradiated tissue, and subsequently performing ultrasonic irradiation during a second time period using unfocused ultrasonic energy at a second intensity level, wherein the second intensity level is lower than the first intensity level.

[0035] According to one aspect of some embodiments of the present disclosure, a procedure for controlling an ultrasonic source is provided, the procedure comprising ultrasonic irradiation with ultrasonic energy of a first intensity level during a first time period, and ultrasonic irradiation with ultrasonic energy of a second intensity level during a second time period, wherein the first intensity level is higher than the second intensity level.

[0036] According to one aspect of some embodiments of this disclosure, a method for programming an ultrasound source is provided, the method comprising programming the ultrasound source to perform ultrasound irradiation using ultrasound energy of a first intensity level during a first time period, and programming the ultrasound source to perform ultrasound irradiation using ultrasound energy of a second intensity level during a second time period, wherein the first intensity level is higher than the second intensity level.

[0037] The following is a non-exclusive list of some examples including embodiments of the present invention. The invention also includes embodiments that incorporate all features from fewer than one example, and embodiments that use features from multiple examples, although not explicitly listed below.

[0038] Example 1. A method for inducing the shedding of cells and molecules from a target epithelium, the method comprising:

[0039] High-energy focused ultrasound with a mechanical index (MI) greater than 0.4 is used to irradiate the target tissue to generate shear waves within the tissue.

[0040] Example 2. The method according to Example 1, wherein the ultrasonic irradiation comprises ultrasonically irradiating a target tissue with ultrasonic energy having a focused beam, the focused beam generating shear waves within the tissue.

[0041] Example 3. The method according to Example 1 or 2, wherein the ultrasonic irradiation includes applying a focused ultrasonic beam, wherein the ultrasonic beam is arranged as follows:

[0042] The mechanical index (MI) is generated in the range of 0.4 to 1.8;

[0043] Produces a focused beamwidth in the range of 2 mm to 10 mm / Produces a beamwidth in the range of 2 mm to 10 mm at the focal point;

[0044] Generated with pulse widths ranging from 100 microseconds to 800 microseconds; and

[0045] It is generated at a pulse repetition rate in the range of 4 Hz to 20 Hz.

[0046] Example 4. The method according to any one of Examples 1-3, wherein ultrasonic irradiation comprises:

[0047] The method further includes irradiating the target tissue with ultrasound energy at a first intensity level during a first time period, and wherein the method also includes irradiating the target tissue with ultrasound energy at a second intensity level during a second time period, wherein the first intensity level is higher than the second intensity level.

[0048] Example 5. The method according to Example 4, wherein the first intensity level corresponds to a mechanical index (MI) in the range of 0.3 to 1.8.

[0049] Example 6. The method according to any one of Examples 4-5, wherein irradiating the target tissue with ultrasound energy during the first time period comprises irradiating with ultrasound via a focused ultrasound beam.

[0050] Example 7. The method according to any one of Examples 4-6, wherein the second intensity level corresponds to a mechanical index (MI) in the range of 0.1 to 0.4.

[0051] Example 8. The method according to any one of Examples 4-7, wherein irradiating the target tissue with ultrasound energy during the second time period comprises irradiating with ultrasound via a non-focused ultrasound beam.

[0052] Example 9. The method according to any one of Examples 4-8, wherein irradiating the target tissue with ultrasound energy during a first time period comprises irradiating with ultrasound pulses of repetitive pulse width in the range of 2 microseconds to 800 microseconds.

[0053] Example 10. The method according to any one of Examples 4-9, wherein irradiating the target tissue with ultrasound energy during the second time period comprises irradiating with ultrasound pulses of repeated duration in the range of 20 microseconds to 800 microseconds.

[0054] Example 11. The method according to any one of Examples 9-10, wherein irradiating the target tissue with ultrasound energy during a first time period comprises repeating ultrasound pulses at a rate ranging from 1 to 20 times per second.

[0055] Example 12. The method according to any one of Examples 9-11, wherein irradiating the target tissue with ultrasound energy during the second time period comprises repeating ultrasound pulses at a rate ranging from 1 to 100 times per second.

[0056] Example 13. The method according to any one of Examples 1-11, wherein the duration of the first time period is in the range of 1 minute to 15 minutes.

[0057] Example 14. The method according to any one of Examples 1-13, wherein the duration of the second time period is in the range of 3 minutes to 30 minutes.

[0058] Example 15. The method according to any one of Examples 4-12, wherein the ultrasound contrast agent is provided to the target tissue prior to ultrasound irradiation of the target tissue with ultrasound energy at a second intensity level during the second time period.

[0059] Example 16. The method according to any one of Examples 4-15, wherein irradiating the target tissue with ultrasound energy during the second time period includes providing an ultrasound contrast agent to the target tissue during the process of irradiating the target tissue with ultrasound energy during the second time period.

[0060] Example 17. The method according to any one of Examples 4-16, wherein the duration of the second time period is in the range of 10 minutes to 20 minutes.

[0061] Example 18. The method according to any one of Examples 4-16, wherein the target tissue includes the pancreas.

[0062] Example 19. The method according to Example 18 further includes: injecting a patient with a drug that induces pancreatic secretion; and collecting cytopathological samples by collecting pancreatic secretions.

[0063] Example 20. The method according to any one of Examples 4-18, wherein the target tissue comprises body parts selected from:

[0064] mediastinum;

[0065] pleura;

[0066] Pericardium;

[0067] peritoneum;

[0068] lung;

[0069] breast;

[0070] Salivary glands;

[0071] Meninges;

[0072] pancreas;

[0073] Pancreatic duct;

[0074] Pancreatic cysts;

[0075] prostate;

[0076] kidney;

[0077] liver;

[0078] Bladder; and

[0079] Ovary.

[0080] Example 21. The method according to Example 20, wherein the target tissue includes the prostate, and wherein the method includes collecting one or more of urine and semen.

[0081] Example 22. The method according to Example 20 or 21 includes promoting urination in a patient by administering a diuretic substance to the patient and inducing the patient to consume fluids in one or more ways.

[0082] Example 23. A method for inducing exfoliation, the method comprising irradiating a target tissue with ultrasonic energy having a mechanical index (MI) in the range of 1.2 to 1.8.

[0083] Example 24. The method according to Example 23, wherein irradiating the target tissue with ultrasound energy comprises irradiating with ultrasound via a focused ultrasound beam.

[0084] Example 25. The method according to any one of Examples 23-24, wherein irradiating the target tissue with ultrasound energy comprises irradiating with ultrasound pulses of repetitive pulse width in the range of 20 microseconds to 800 microseconds.

[0085] Example 26. The method according to Example 25, wherein irradiating the target tissue with ultrasound energy comprises repeating ultrasound pulses at a rate of 1 to 50 times per second.

[0086] Example 27. An ultrasonic energy source programmed to perform ultrasonic irradiation using an ultrasonic irradiation procedure, the procedure comprising:

[0087] Select program parameters to generate shear waves used to induce or facilitate cell shearing; and

[0088] During the first time period, focused ultrasound energy designed to generate shear waves in the irradiated tissue was used for ultrasound irradiation at a first intensity level.

[0089] Example 28. An ultrasonic energy source according to Example 27, wherein, after or before the ultrasonic irradiation:

[0090] During the second time period, ultrasound irradiation was performed using unfocused ultrasound energy at a second intensity level, where the second intensity level was lower than the first intensity level.

[0091] Example 29. A program for controlling an ultrasound source, the program comprising:

[0092] During the first time period, ultrasonic irradiation was performed using ultrasonic energy of the first intensity level.

[0093] Subsequently

[0094] During the second time period, ultrasonic irradiation was performed using ultrasonic energy at the second intensity level.

[0095] in

[0096] The first intensity level is higher than the second intensity level.

[0097] Example 30. A method for programming an ultrasound source, the method comprising:

[0098] The ultrasound source is programmed to perform ultrasound irradiation using ultrasound energy of a first intensity level during a first time period.

[0099] The ultrasound source is programmed to perform ultrasound irradiation using a second intensity level of ultrasound energy during the second time period.

[0100] in

[0101] The first intensity level is higher than the second intensity level.

[0102] Example 31. A separated sample of bodily fluid obtained from an ultrasound-irradiated prostate of a subject, wherein the separated sample contains exfoliated cells in an amount sufficient for the analysis and detection of abnormal cells.

[0103] Example 32. A separated sample of bodily fluids according to Example 31, comprising one or more of urine and semen.

[0104] Example 33. A separated sample of bodily fluid according to Example 31 or 32, comprising adjacent cells from the prostate.

[0105] Example 34. A separated sample of body fluid according to Example 33, wherein the adjacent cells comprise fragments or sheets of epithelial tissue.

[0106] Example 35. A separated sample of body fluid according to Example 34, containing more than one fragment of epithelial tissue.

[0107] Example 36. A separated sample of body fluid according to Example 34 or 35, comprising intact epithelial tissue fragments from an organ.

[0108] Example 37. A separated sample of bodily fluid according to any one of the preceding examples, wherein the separated sample is obtained by irradiating the prostate with high-intensity focused ultrasound.

[0109] Example 38. An ultrasound device programmed to irradiate the prostate with ultrasound, comprising at least one probe for irradiating the prostate with ultrasound, wherein the at least one probe comprises one or more of the following: a transrectal ultrasound probe, a transperineal ultrasound probe, and an ultrasound catheter probe.

[0110] Example 39. An ultrasound device programmed to irradiate the prostate with ultrasound according to Example 38, including an electroejaculation device.

[0111] Example 40. An ultrasound device programmed to irradiate the prostate with ultrasound according to Example 38 or 39, including a prostate massager.

[0112] Example 41. An ultrasound device programmed to irradiate the prostate with ultrasound according to any one of Examples 38-39, is programmed to:

[0113] Select program parameters to generate shear waves used to induce or facilitate cell shearing; and

[0114] During the first time period, focused ultrasound energy designed to generate shear waves in the irradiated tissue was used for ultrasound irradiation at a first intensity level.

[0115] Example 42. An ultrasound device programmed to irradiate the prostate with ultrasound according to Example 41, wherein, after or before the ultrasound irradiation:

[0116] During the second time period, ultrasound irradiation was performed using unfocused ultrasound energy at a second intensity level, where the second intensity level was lower than the first intensity level.

[0117] Example 43. A method for inducing exfoliation, comprising irradiating a target tissue with ultrasound energy in the range of 1.2 to 1.8 by means of an ultrasonic index (MI).

[0118] Example 44. The method according to Example 43, wherein irradiating the target tissue with ultrasound energy comprises irradiating with ultrasound via a focused ultrasound beam.

[0119] Example 45. The method according to any one of Examples 43-44, wherein irradiating the target tissue with ultrasound energy comprises irradiating with ultrasound pulses of repetitive pulse width in the range of 20 microseconds to 800 microseconds.

[0120] Example 46. The method according to Example 45, wherein irradiating the target tissue with ultrasound energy comprises repeating ultrasound pulses at a rate of 1 to 50 times per second.

[0121] Unless otherwise defined, all technical and / or scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this disclosure pertains. While methods and materials similar to or equivalent to those described herein may be used in practice or testing of embodiments of this disclosure, exemplary methods and / or materials are described below. In case of conflict, the patent specification (including definitions) shall prevail. Furthermore, materials, methods, and examples are illustrative only and are not intended to be restrictive.

[0122] As those skilled in the art will understand, some embodiments of this disclosure may be embodied as a system, method, or computer program product. Therefore, some embodiments of this disclosure may take the form of a completely hardware implementation, a completely software implementation (including firmware, resident software, microcode, etc.), or an implementation combining software and hardware aspects, which are generally referred to herein as “circuit,” “module,” or “system.” Furthermore, some embodiments of this disclosure may take the form of a computer program product contained in one or more computer-readable media, on which computer-readable program code is contained. Implementation of the methods and / or systems of some embodiments of this disclosure may include performing and / or completing selected tasks manually, automatically, or in combination thereof. Furthermore, according to the actual instruments and equipment of some embodiments of the methods and / or systems of this disclosure, several selected tasks may be implemented by hardware, software, or firmware and / or a combination thereof, such as using an operating system.

[0123] For example, according to some embodiments of this disclosure, the hardware for performing a selected task can be implemented as a chip or circuit. As software, the selected task according to some embodiments of this disclosure can be implemented as a plurality of software instructions executed by a computer using any suitable operating system. In exemplary embodiments of this disclosure, one or more tasks of some exemplary embodiments of the methods and / or systems described herein are performed by a data processor, such as a computing platform for executing multiple instructions. Optionally, the data processor includes volatile memory for storing instructions and / or data and / or non-volatile memory for storing instructions and / or data, such as a magnetic hard disk and / or removable media. Optionally, a network connection is also provided. A display and / or user input device, such as a keyboard or mouse, may also be optionally provided.

[0124] Any combination of one or more computer-readable media can be used in some embodiments of this disclosure. A computer-readable medium can be a computer-readable signal medium or a computer-readable storage medium. A computer-readable storage medium can be, for example, but not limited to, electronic, magnetic, optical, electromagnetic, infrared, or semiconductor systems, devices, or apparatuses, or any suitable combination of the foregoing. More specific examples (not an exhaustive list) of computer-readable storage media will include the following: an electrical connection having one or more wires, a portable computer disk, a hard disk, random access memory (RAM), read-only memory (ROM), erasable programmable read-only memory (EPROM or flash memory), optical fiber, portable optical disc read-only memory (CD-ROM), optical storage device, magnetic storage device, or any suitable combination of the foregoing. In the context of this document, a computer-readable storage medium can be any tangible medium capable of containing or storing a program used by or in connection with an instruction execution system, apparatus, or device.

[0125] Computer-readable signal media may include propagated data signals containing computer-readable program code, for example, in baseband or as part of a carrier wave. Such propagated signals may take any of a variety of forms, including, but not limited to, electromagnetic, optical, or any suitable combination thereof. A computer-readable signal medium may be any computer-readable medium that is not a computer-readable storage medium and may transmit, propagate, or transfer a program used by or in connection with an instruction execution system, apparatus, or device.

[0126] The program code contained on a computer-readable medium and / or the data used therefrom can be transmitted using any suitable medium, including but not limited to wireless, wired, fiber optic cable, RF, or any suitable combination thereof.

[0127] Computer program code used to perform operations of some embodiments of this disclosure may be written in any combination of one or more programming languages, including object-oriented programming languages ​​such as Java, Smalltalk, and C++, and traditional procedural programming languages ​​such as the "C" programming language or similar programming languages. The program code may execute entirely on the user's computer, partially on the user's computer, as a standalone software package, partially on the user's computer and partially on a remote computer, or entirely on a remote computer or server. In the latter case, the remote computer may be connected to the user's computer via any type of network, including a local area network (LAN) or a wide area network (WAN), or may be connected to an external computer (e.g., via the Internet through an Internet service provider).

[0128] Some embodiments of this disclosure may be described below with reference to flowchart illustrations and / or block diagrams of methods, apparatus (systems), and computer program products according to embodiments of this disclosure. It will be understood that each block of the flowchart illustrations and / or block diagrams, and combinations of blocks in the flowchart illustrations and / or block diagrams, can be implemented by computer program instructions. These computer program instructions can be provided to a processor of a general-purpose computer, special-purpose computer, or other programmable data processing apparatus to produce a machine, such that the instructions, which execute via the processor of the computer or other programmable data processing apparatus, create mechanisms for implementing the functions / actions specified in one or more blocks of the flowchart illustrations and / or block diagrams.

[0129] These computer program instructions may also be stored in a computer-readable medium that can instruct a computer, other programmable data processing device or other apparatus to operate in a particular manner, such that the instructions stored in the computer-readable medium produce an article of writing comprising instructions that implement the functions / actions specified in one or more boxes of a flowchart and / or block diagram.

[0130] Computer program instructions may also be loaded onto a computer, other programmable data processing apparatus or other device to cause a series of operational steps to be performed on the computer, other programmable apparatus or other device, thereby producing a computer-implemented process, such that the instructions, which execute on the computer or other programmable apparatus, provide a process for implementing the functions / actions specified in one or more boxes of a flowchart and / or block diagram.

[0131] Some of the methods described herein are typically designed for computer use only and may be infeasible or impractical for human experts to perform entirely manually. Human experts who wish to perform similar tasks manually (e.g., performing a combination of ultrasound methods to enhance cell collection in cytopathology) may be expected to use entirely different approaches, such as leveraging expert knowledge and / or the pattern recognition capabilities of the human brain, which may be more efficient than manually going through the steps of the methods described herein. Attached Figure Description

[0132] Some embodiments of this disclosure are described herein by way of example only with reference to the accompanying drawings and images. Reference will now be made in detail to the accompanying drawings and images, and it is emphasized that the details shown are exemplary and intended for illustrative discussion of embodiments of this disclosure. In this respect, the description taken in conjunction with the drawings will make it clear to those skilled in the art how to implement embodiments of this disclosure.

[0133] In the attached diagram:

[0134] Figure 1A This is a simplified qualitative illustration of ultrasound irradiation for inducing detachment of cell samples in cytopathology, according to an exemplary embodiment of the present invention.

[0135] Figure 1B This is a simplified qualitative illustration of ultrasound irradiation during a process for producing cytopathological cell samples, according to an exemplary embodiment of the present invention.

[0136] Figure 1C This is a simplified qualitative illustration of the level of ultrasound irradiation during a process for producing cytopathological cell samples according to an exemplary embodiment of the present invention.

[0137] Figure 1D This is a simplified qualitative illustration of the level of ultrasound irradiation during a process for producing cytopathological cell samples according to an exemplary embodiment of the present invention.

[0138] Figure 2A This is a simplified flowchart illustrating a method for generating cytopathological cell samples according to an exemplary embodiment of the present invention;

[0139] Figure 2BThis is a simplified flowchart illustrating a method for generating cytopathological cell samples according to an exemplary embodiment of the present invention;

[0140] Figure 2C This is a simplified flowchart illustrating a method for generating cytopathological cell samples according to an exemplary embodiment of the present invention;

[0141] Figure 2D This is a simplified flowchart of a method for inducing exfoliation according to an exemplary embodiment of the present invention;

[0142] Figures 3A-3D This is a simplified illustration of using ultrasound and an ultrasound contrast agent to generate a cell sample according to an exemplary embodiment of the present invention;

[0143] Figure 3E This is a simplified illustration of using ultrasound and an ultrasound contrast agent to generate a cell sample according to an exemplary embodiment of the present invention;

[0144] Figure 4A This is a front perspective view of an ultrasonic probe according to one embodiment of the present invention;

[0145] Figure 4B This is a schematic diagram of a probe configuration according to an exemplary embodiment of the present invention;

[0146] Figure 4C This is a simplified diagram of an array of elements in an ultrasonic transducer according to an exemplary embodiment of the present invention;

[0147] Figure 4D This is a simplified diagram of an array of elements in an ultrasonic transducer according to an exemplary embodiment of the present invention;

[0148] Figure 5 This is an image of a set of adjacent cells obtained through an exemplary embodiment of the present invention;

[0149] Figure 6 This is a flowchart of a method for generating cytopathological cell samples from the prostate according to an exemplary embodiment of the present invention;

[0150] Figure 7 This is a schematic cross-sectional view of a prostatic sac (or microgland) and a segment of a prostatic duct according to an exemplary embodiment of the present invention;

[0151] Figure 8 These are schematic cross-sectional views of the patient's pelvic region and simplified diagrams of an ultrasound probe interacting with the patient's prostate, according to various exemplary embodiments of the present invention; and

[0152] Figure 9This is a table showing the experimental results of the embodiments of the present invention on human patients. Detailed Implementation

[0153] This disclosure relates, in some embodiments thereof, to an ultrasound method for breaking up target epithelium using ultrasonic shear waves, to an ultrasound device constructed and / or programmed to use this method, and to programming an ultrasound device to use this method.

[0154] This disclosure relates, in some embodiments thereof, to an ultrasound method for enhancing the stripping of cells and / or molecules of a target epithelium, to constructing and / or programming an ultrasound device for using this method, and to programming an ultrasound device to use this method.

[0155] A broad aspect of some embodiments of the invention includes irradiating tissue with ultrasound over a period of time to potentially induce shear waves in the tissue, thereby promoting cell detachment from the tissue, optionally by promoting separation of cells and / or tissue fragments from the tissue and / or promoting separation between cells and / or tissue fragments. Note that in some embodiments, detachment is preferably induced without causing thermal and / or persistent damage to the tissue / organ. It should also be noted that, typically, cells and tissues regenerate after detachment. In some embodiments, detachment produces a sample containing unruptured cells and / or intact tissue fragments in sufficient quantity for detecting abnormalities.

[0156] In some implementations, shear waves are induced by applying ultrasound, optionally high-intensity focused ultrasound. Unbound by theory, the target epithelium can potentially be segmented by creating transverse breaks (e.g., disconnections) between cells. Unbound by theory, generating shear waves in tissues / organs potentially tends to separate aberrations (e.g., cancer cells and / or tissue fragments) from the organ and / or separate them from each other, which has the potential advantage of enhanced sample reliability.

[0157] A broad aspect of some embodiments of the invention includes irradiating tissue with high-intensity focused ultrasound during a first time period to generate shear waves that partially segment the target epithelium by creating transverse breaks between cells and / or their bases, followed by injection of a microbubble contrast agent, and irradiating the tissue with lower-intensity ultrasound during a second time period, which is not theoretically constrained, to separate the partially segmented epithelium from the organ surface, thereby inducing the shedding of cell and tissue debris.

[0158] A broad aspect of some embodiments of the invention includes irradiating tissue with ultrasound of lower intensity during a first time period, followed by irradiating the tissue with ultrasound of higher intensity during a second time period to induce cell detachment from the tissue. In some embodiments, the higher intensity ultrasound during the second time period is focused ultrasound. In some embodiments, a microbubble contrast agent is injected prior to irradiating the tissue with lower intensity ultrasound during the first time period. Not bound by theory, in some embodiments, irradiating microbubbles with lower intensity ultrasound promotes the separation of tissue debris from the organ, while the higher intensity ultrasound promotes the separation of cells from the organ. This sequence has the potential advantage of increasing the number and / or size of detached cell populations and / or tissue debris (e.g., flakes).

[0159] Overview

[0160] Some of the methods and systems described herein are used to obtain cells and tissue fragments detached from target tissues in greater quantities than previously observed by different means through non-limiting example epithelium, and may also obtain biomolecules related to developmental abnormalities and / or cancer that may also detach from target tissues.

[0161] Some of the methods and systems described herein are used to obtain adjacent cell and / or cell population, cell sheet, and / or tissue fragment samples at a higher rate (i.e., number of samples per unit time) than previously observed by different means, for the purpose of evaluating the structure of epithelium from target tissues.

[0162] Some of the methods and systems described herein are used to obtain cell slices, and / or cell populations and / or tissue fragments, which contain a greater number of cells in the slices / populations / fragments than previously observed by different means.

[0163] For example, target tissues such as the pancreas may be irradiated with ultrasound at a higher intensity during a first time period, followed by irradiation with ultrasound at a lower intensity during a second time period. There may or may not be a quiet period between the first and second time periods of ultrasound irradiation.

[0164] For example, target tissues such as the pancreas may be irradiated with ultrasound at a lower intensity during a first time period, and then irradiated with ultrasound at a higher intensity during a second time period. There may or may not be a quiet period between the first and second time periods of ultrasound irradiation.

[0165] Shear waves are induced by high-intensity, long-duration (e.g., 100 to 800 microseconds) focused ultrasound beam pulses. Typically, this ultrasound beam is perpendicular to the surface of the epithelium. The ultrasound beam that induces shear waves typically travels through the organ and generates pressure changes that cause rapid expansion and contraction, usually parallel to the surface plane of the tissue. These mechanical changes can potentially cause dissociation in the lateral / transverse connections between epithelial cells. Note that standard imaging ultrasound uses a pulse width of 2 microseconds.

[0166] Ultrasound contrast agents can be gas microbubbles ranging in size from 1 μm to 4 μm, optionally encapsulated in protein, lipid, or polymer shells, which expand and contract upon exposure to ultrasound due to changes in their local pressure. Therefore, when intravenously injected into a subject, these microbubbles flow through the vascular system and all its capillaries, reaching the rich microvascular system surrounding the acinar cells and ductal system. The combined effect of the ultrasound beam aimed at the pancreas and the presence of microbubbles within its microvascular system potentially causes synchronous expansion and contraction of the microvascular system beneath the epithelial layer, thereby displacing cells and cell clusters by disrupting the cellular connections between the basement membrane of the epithelial cells.

[0167] In some examples, the ultrasound irradiation in the first time period includes ultrasound irradiation via focused ultrasound energy, which potentially generates shock waves in the tissue and / or potentially induces shear waves in the tissue. In some examples, the higher energy ultrasound energy in the first time period corresponds to a mechanical index (MI) in the range of 1.0 to 1.8.

[0168] In some examples, the ultrasound irradiation during the second time period includes ultrasound irradiation with unfocused ultrasound energy. In some examples, the lower energy ultrasound energy during the second time period corresponds to an MI in the range of 0.1 to 0.4 or even 1.0. As a non-limiting example, the MI could be 0.25 or 0.3.

[0169] In some examples, ultrasound contrast material, such as Lumison microbubbles as a non-limiting example, is provided at the target tissue before and / or during ultrasound irradiation in a second time period. The ultrasound contrast material interacts with ultrasound energy and can be used to remove cell and / or cell sheet samples, and can be used to transport samples along the lumen of the target tissue to be collected.

[0170] Various non-limiting examples of ultrasound contrast materials include:

[0171] The Lumison mentioned above;

[0172] Optison, a microbubble approved by the U.S. Food and Drug Administration (FDA) and supplied by GE Healthcare, features an albumin shell and an octafluoropropane gas core.

[0173] Definity is an FDA-approved microbubble supplied by Lantheus. Definity comprises perfluorolipid microspheres, each consisting of octafluoropropane encapsulated in a lipid shell; and

[0174] Lumason, also known as SonoVue, is supplied by Bracco and is FDA approved. It is a lipid-encapsulated microvesicle containing sulfur hexafluoride.

[0175] In some examples, ultrasound irradiation for the first time period and / or ultrasound irradiation for the second time period includes ultrasound irradiation using ultrasound pulses with a width or pulse duration of 200 microseconds. In some examples, ultrasound irradiation for the first time period and / or ultrasound irradiation for the second time period includes ultrasound irradiation using ultrasound pulses with a width or pulse duration in the range of 20 microseconds to 800 microseconds (as a non-limiting example, a pulse width of 200 microseconds).

[0176] In some examples, ultrasound irradiation of the first time period and / or ultrasound irradiation of the second time period includes ultrasound irradiation by repeating ultrasound pulses at a relatively high repetition rate of 8 times per second. In some examples, ultrasound irradiation of the first time period and / or ultrasound irradiation of the second time period includes ultrasound irradiation by repeating ultrasound pulses at a relatively high repetition rate of 1 to 100 times per second (as a non-limiting example, a repetition rate of 20 times per second (20 Hz)).

[0177] Note that in current practice, when ultrasound shear waves are used for various purposes, they are generated at a repetition rate of 1 Hz or 1 second. Increasing the repetition rate allows more energy to be delivered to the irradiated tissue, potentially inducing the separation of intact tissue fragments from the target organ.

[0178] In some examples, the first time period of ultrasound irradiation consists of 5 minutes of shear waves with a higher MI, followed by a second time period of ultrasound irradiation consisting of approximately 10-20 minutes of microbubbles with a lower MI.

[0179] In some examples, the initial ultrasound irradiation lasted from 1 minute to 15 minutes.

[0180] In some examples, the second ultrasound irradiation period lasted 10 to 20 minutes. In other examples, the second ultrasound irradiation period lasted 3 to 30 minutes.

[0181] Cell samples can optionally be collected after ultrasound irradiation by extracting bodily fluids that have come into contact with the tissues of the subject irradiated by ultrasound.

[0182] In some embodiments, the two time periods may optionally be provided in the reverse order described above. In some embodiments, the ultrasound irradiation of the first time period uses unfocused, lower-intensity ultrasound waves, while the ultrasound irradiation of the second time period uses higher-intensity, focused ultrasound energy, which may generate shock waves in the tissue and / or may induce shear waves in the tissue. Optionally, the pulse width, repetition rate, and MI values ​​used in the ultrasound irradiation of the first time period using unfocused, lower-intensity ultrasound waves are similar to those described in the ultrasound irradiation of the second time period using unfocused, lower-intensity ultrasound waves. Optionally, the pulse width, repetition rate, and MI values ​​used in the ultrasound irradiation of the second time period using focused, higher-intensity ultrasound waves are similar to those of the focused, higher-intensity ultrasound waves used in the ultrasound irradiation of the first time period described above, which may generate shock waves in the tissue and / or may induce shear waves in the tissue.

[0183] In some implementations, secretin is used to induce pancreatic juice secretion. Secretin requires 10-15 minutes to reach its maximum effect. Both secretin and ultrasound contrast agents are administered intravenously and typically require a caregiver to administer. Shear wave ultrasound irradiation generally does not require a caregiver and / or intravenous injection. In other target organs, depending on the nature of the organ, secretin can be replaced by another drug that promotes flushing. For example, salivary enhancement materials for salivary glands.

[0184] In some implementations, procedures requiring caregivers, such as administration of ultrasound contrast agents and low-intensity non-focused ultrasound irradiation, along with the administration of secretin, may optionally be performed simultaneously. In some implementations, the drug administration and the low-intensity non-focused ultrasound are combined and performed first, prior to the high-intensity focused ultrasound.

[0185] The duration of non-focused, low-intensity ultrasound irradiation described in this article can typically last about 10-15 minutes, similar to the duration required for secretion to reach its maximum effect, which potentially provides the beneficial effect of performing both simultaneously.

[0186] Acoustic hole effect

[0187] One aspect of some implementation schemes involves using ultrasound irradiation, as described herein, to destroy tissue, which could potentially enhance drug delivery.

[0188] In some implementations, as described herein, lower intensity ultrasound irradiation combined with microbubbles generated by ultrasound contrast agents to disrupt tissue can potentially enhance drug delivery.

[0189] In some implementations, such as those described herein, higher intensity ultrasound irradiation is used to destroy tissue, which can potentially enhance drug delivery.

[0190] In some implementations, higher-intensity ultrasound irradiation, as described herein, followed by lower-intensity ultrasound irradiation, as described herein, plus microbubbles generated by an ultrasound contrast agent to destroy tissue, can potentially enhance drug delivery.

[0191] In some implementations, the tissue may be destroyed by using the reverse sequence of ultrasound irradiation time periods, which could potentially enhance drug delivery.

[0192] target organs

[0193] As described herein, a characteristic of target organs suitable for using ultrasound methods to enhance the collection of cytopathological cells includes organs in which fluids in contact with the epithelium of the ultrasound-irradiated subject have a natural anatomical outlet for fluid collection. A non-limiting example is a glandular organ with an outlet. Some non-limiting examples include pancreatic juice flowing from the ampulla of Vater, mammary duct fluid flowing from the nipple, urine flowing from the kidney and bladder at the urethra, and prostatic fluid flowing from the urethra.

[0194] As described in this article, some non-limiting examples of using ultrasound to enhance target organs in cytopathological cell collection include:

[0195] pancreas;

[0196] Pancreatic duct;

[0197] Pancreatic cysts;

[0198] breast;

[0199] prostate;

[0200] mediastinum;

[0201] pleura;

[0202] Pericardium;

[0203] peritoneum;

[0204] lung;

[0205] Salivary glands;

[0206] Meninges;

[0207] kidney;

[0208] liver;

[0209] Bladder; and

[0210] Ovary.

[0211] Note that many of the exemplary embodiments provided herein are written with the pancreas as an example; however, these examples are not intended to be limited to the pancreas, but rather extend to at least the other target organs listed herein, as well as other organs having the characteristics listed in the first paragraph of the “Target Organs” section.

[0212] For example, in some embodiments, the target organ is the bladder. In some embodiments, the bladder should be full (e.g., with urine) and / or substantially full during ultrasound treatment to potentially accelerate sample collection with less and / or no external intervention. Alternatively or additionally, the patient is prompted to drink a beverage and / or given a diuretic to induce urination. In some embodiments, optionally, the bladder is externally irradiated with ultrasound, with necessary modifications, using a device for the prostate as described herein. For example, an ultrasound probe is applied to the skin in the patient's lower abdominal region and / or perineum. In some embodiments, a bladder-sized patch may be used, optionally, a variant of probe 410 (e.g., in...) Figure 4A (As shown in the diagram) it has a larger probe contact surface. In some embodiments, the device includes a strap (e.g., strap 414) whose size and / or shape are adjusted to fit the bladder. For example, a probe for the perineum is attached to a strap whose size and / or shape are adapted to the pelvic region.

[0213] In some implementations, irradiating the bladder with ultrasound to generate shear waves in the tissue (as described herein) has the potential advantage of reducing the number and / or duration of microbubbles, which potentially reduces and / or prevents microbubble interactions with other (e.g., surrounding) organs. Furthermore, generating shear waves at the bladder potentially allows ultrasound waves to be focused at the bladder, with the potential advantage of reducing and / or avoiding ultrasound damage to surrounding organs, blood vessels, and / or nerves.

[0214] motivation

[0215] The ultrasound application described herein can be described as irradiating tissue with ultrasound in two phases: 1) generating shear waves at the target tissue, optionally, focused shear waves. In some embodiments, shear waves in the tissue are generated by applying focused ultrasound with a high MI (Minimum Intensity) without microbubbles, for example, with an MI in the range of 1.2–1.8, or 1–1.9, or 0.5–1.7, or with an MI of about 1.4, or about 1, or about 2, or about 1.6, or a lower or higher MI, or an intermediate value. Currently, the FDA approves MIs not exceeding 1.9, and in some embodiments, higher MI values ​​(e.g., greater than 1.9) can be selected because the ultrasound is not used for imaging.

[0216] Unbound by theory, this causes the epithelium to vibrate in a plane roughly parallel to the basement membrane, disrupting the transverse bonds / side bonds between epithelial cells, followed by: 2) low-intensity unfocused ultrasound irradiation with an MI equal to or less than 0.4, which potentially leads to microbubble resonance introduced into the veins beneath the basement membrane. As a result, tissue clusters whose side bonds were disrupted in stage 1 now have their basement bonds disrupted, causing cells and large tissue clusters to be lifted from below. In some embodiments, contrast agent microbubbles are made to oscillate within the vascular system supplying the epithelial layer.

[0217] It is believed that the procedures / processes described herein can potentially significantly increase the total number of pancreatic cells, such as those in pancreatic juice, for expression. Furthermore, the procedures described herein can induce the separation of intact tissue fragments from target organs such as the pancreas.

[0218] When applied to other organs or body parts, such as, for example, the mediastinum, pleura, pericardium, peritoneum, lungs, breast, salivary glands, meninges, pancreatic duct, pancreatic cysts, kidneys, liver, bladder, or ovaries, such a sequence can significantly increase the total number of cells expressed in the fluid surrounding these organs.

[0219] In some implementations, the procedures described herein can potentially induce the separation of intact tissue fragments from the aforementioned exemplary organs. For example, the procedures described herein can optionally be used to induce lung cells to detach into surrounding sputum / saliva, or to induce bladder cells and bladder tissue to detach into surrounding urine.

[0220] One aspect of some embodiments of the present invention relates to non-invasive early detection of pancreatic cancer and / or developmental abnormalities. Pancreatic cancer is an extremely deadly cancer, primarily because it is often diagnosed at a late stage.

[0221] This invention provides a system and method for inducing pancreatic cell shedding to obtain sufficient cells for the analysis and detection of abnormal cells. For example, some existing conventional pancreatic juice sampling techniques typically yield fewer than 50 pancreatic cells and fail to collect intact tissue fragments, while the embodiments described herein can reliably obtain samples containing more than 100 pancreatic cells while capturing intact tissue sections. In some embodiments, histological analysis can be performed. Evaluation of the tissue (optionally intact tissue sections) allows for the assessment of the honeycomb pattern of cells characterizing benign glandular epithelium, while the detection of the disappearance of this honeycomb pattern characterizes developmental abnormalities and cancer.

[0222] One aspect of some embodiments of the present invention relates to non-invasive early detection of prostate cancer and / or developmental abnormalities, performed by testing isolated samples of urine and / or semen containing prostate cells and / or tissue from a patient's prostate. In some embodiments, the prostate is irradiated with ultrasound to generate shear waves. In an unconstrained manner, the shear waves partially segment the target epithelium by creating transverse breaks between cells and / or cell bases, optionally, as described herein, by applying high-intensity focused ultrasound. Exfoliated cells and tissue from the prostate are discharged into the urethra and expelled during ejaculation and / or urination, and are collected for analysis. Furthermore, exfoliated cells from the urethra may enter the bladder (e.g., retrogradely into the bladder and then expelled with the subject's urine) and are collected for analysis.

[0223] Early detection of prostate cancer can potentially increase the chances of successful treatment. However, because this disease is often asymptomatic before it progresses to an advanced stage, it is frequently detected at a late stage, when the cancer is debilitating and / or life-threatening. Embodiments of the present invention potentially allow for enhanced and / or accelerated expulsion of prostate cells and / or tissue, thereby making urine and / or semen collection a viable and / or non-invasive option for early detection of prostate cancer and precancerous conditions. Urine and / or semen collection has the potential advantage of minimally invasive testing. In some embodiments, urine and / or semen sample collection can be used for prostate cancer screening and / or prostate cancer monitoring (e.g., monitoring patients diagnosed with prostate cancer but not receiving aggressive treatment, or patients with abnormal cells in previously diagnosed samples, or patients diagnosed with prostate cancer who are receiving treatment, in order to monitor treatment success and / or disease progression, optionally collecting samples from patients at a certain frequency, such as weekly or monthly, which optionally depends on the state of the disease).

[0224] Embodiments of the present invention provide a system and method for inducing the separation of urine and / or semen samples, said separated samples comprising detached prostate cells in a sufficient number for analysis and detection of abnormal cells. In embodiments of the present invention, cells detach from the prostate at a greater number and / or rate, and comprise cell clumps, compared to a prostate not irradiated with ultrasound and / or not irradiated with relatively high-intensity focused ultrasound to generate shear waves.

[0225] Some of the methods and systems described herein are used to obtain samples comprising at least one of the following detached from the prostate: cells, adjacent / neighboring cells, cell sheets, cell populations and / or cell clusters, and / or containing at least one tissue fragment detached from the prostate. In some embodiments, the sample comprises intact tissue sheets and / or intact tissue fragments.

[0226] Ultrasound irradiation of the prostate to generate shear waves potentially allows for the reduction and / or avoidance of the mechanical effects of ultrasound irradiation on the exterior and / or interior of the prostate, particularly on microbubbles. This has the potential advantage of reducing and / or avoiding undesirable interactions between microbubbles and their surrounding environment, organ nerves, and / or blood vessels. For example, it could reduce and / or avoid the risk of affecting the bladder and urethra and / or causing erectile dysfunction, for example, due to microbubbles. Furthermore, ultrasound irradiation of the prostate to generate shear waves potentially allows for the focusing of ultrasound waves onto the target tissue / organ, with the potential advantage of reducing and / or avoiding interactions between ultrasound waves and surrounding organs, nerves, and / or blood vessels.

[0227] In some embodiments, prostate stimulation is performed to potentially promote prostate fluid secretion. In some embodiments, stimulation includes sexual stimulation, prostate massage, and / or electroejaculation (e.g., delivering an electric current to the prostate).

[0228] In embodiments of the invention, urination is promoted in the patient. In some embodiments, at least one diuretic substance, such as furosemide or a similar diuretic, may be administered to the patient to increase urine output. Alternatively or additionally, the patient may be prompted to drink a beverage before, after, and / or during ultrasound treatment.

[0229] Preserve the sample

[0230] Once the samples are collected, the cell samples may optionally be preserved under certain conditions and / or processed to minimize autolysis.

[0231] As a non-limiting example, cell samples may optionally be preserved by adding formalin.

[0232] As a non-limiting example, when a pancreatic juice sample contains tissue from the pancreas and the sample is immersed in pancreatic enzymes, the cells of the pancreatic sample in the pancreatic juice may be digested, competing with the cross-linking fixation of the same cells performed by formalin.

[0233] Enzyme activity is inhibited by cold. In some implementations, cell samples may be stored in wide-mouthed flasks on ice or embedded in ice.

[0234] As a non-limiting example, when the sample contains tissue from the pancreas and is immersed in pancreatic enzymes, catalytic activity of the pancreatic sample cells may exist in the pancreatic juice, competing with the cross-linking fixation performed by formalin. Pancreatic enzymes require a relatively alkaline environment; an acidic environment will deactivate them.

[0235] As a non-limiting example, the pH of the formalin or other fixative preservative in which the fluid sample is placed may optionally be lowered.

[0236] As a non-limiting example, the fixative for lowering pH can be formalin, alcohol, or a mixture of both.

[0237] Before explaining at least one embodiment of this disclosure in detail, it should be understood that this disclosure is not necessarily limited in its application to the details of the construction and arrangement of the components and / or methods set forth in the following description and / or shown in the drawings and / or examples. This disclosure can have other embodiments, or can be practiced or implemented in many different ways.

[0238] Before explaining at least one embodiment of this disclosure in detail, it should be understood that this disclosure is not necessarily limited in its application to the details set forth in the following description or illustrated by examples. This disclosure can have other embodiments or can be practiced or implemented in many different ways.

[0239] Now for reference Figure 1A The figure is a simplified qualitative illustration of ultrasound irradiation for inducing the shedding of cytopathological cell samples according to an exemplary embodiment of the present invention.

[0240] Figure 1A An ultrasound source 132 is shown that generates ultrasound waves 134 to enter tissues.

[0241] Figure 1A The acoustic radiation force field from the focused ultrasound beam 134 is shown. The force field, or excitation region, lies within the geometric shadow of the emission aperture and typically has the highest energy near the focal point 136. The high-intensity focused ultrasound beam 134 can be used to agitate tissue to generate shear waves 138a, 138b, which propagate laterally away from the excitation region or focal point 136.

[0242] In some implementations, a focused ultrasound beam may optionally be used to induce the shedding of cells and molecules from a target tissue, such as epithelium, by applying the focused ultrasound beam.

[0243] In some implementations, a focused ultrasound beam is generated to induce a mechanical index (MI) ranging from 0.4 to 1.8.

[0244] In some implementations, the focused ultrasound beam is arranged to produce a beamwidth in the range of 2 mm to 10 mm at the focal point.

[0245] In some implementations, the pulse width generated by the ultrasonic beam is in the range of 100 microseconds to 800 microseconds.

[0246] In some implementations, the pulse repetition frequency generated by the ultrasonic beam is in the range of 4 Hz to 20 Hz.

[0247] Now for reference Figure 1BThe figure is a simplified qualitative illustration of ultrasound irradiation during the process of producing cytopathological cell samples according to an exemplary embodiment of the present invention.

[0248] Figure 1B A graph 140 shows the ultrasound irradiation level over time. Graph 140 has an X-axis 142 for time and a Y-axis 144 for the mechanical index (MI) in the target tissue, which corresponds to the ultrasound irradiation level.

[0249] Figure 1B The ultrasound irradiation during a first time period 146 at a relatively high ultrasound irradiation level is shown as a non-limiting example, where the mechanical index (MI) produced by the ultrasound irradiation level is in the range of 0.4 to 1.8.

[0250] In some implementations, the optional idle time period 147 may exist between the first time period 146 and the optional second time period 148.

[0251] Now for reference Figure 1C , Figure 1C This is a simplified qualitative illustration of the ultrasound irradiation level during the process of generating cytopathological cell samples according to an exemplary embodiment of the present invention.

[0252] Figure 1C A graph 100 shows the ultrasound irradiation level over time. Graph 100 has an X-axis 102 for time and a Y-axis 104 for the mechanical index (MI) in the target tissue, which corresponds to the ultrasound irradiation level.

[0253] As described in this article, Figure 1C The ultrasound irradiation is shown during a first time period 106 at a higher ultrasound irradiation level and during a second time period 108 at a lower ultrasound irradiation level.

[0254] In some implementations, an optional idle time period 107 may exist between the first time period 106 and the second time period 108.

[0255] Now for reference Figure 1D The figure is a simplified qualitative illustration of the ultrasound irradiation level during the process of producing cytopathological cell samples according to an exemplary embodiment of the present invention.

[0256] Figure 1D A graph 120 shows the ultrasound irradiation level over time. Graph 120 has an X-axis 122 for time and a Y-axis 124 for the mechanical index (MI) in the target tissue, which corresponds to the ultrasound irradiation level.

[0257] As described in this article, Figure 1DThe study shows ultrasound irradiation at a lower intensity level during the first time period 126 and ultrasound irradiation at a higher intensity level during the second time period 128.

[0258] In some implementations, an optional idle time period 127 may exist between the first time period 126 and the second time period 128.

[0259] Now for reference Figure 2A The figure is a simplified flowchart of a method for generating cytopathological cell samples according to an exemplary embodiment of the present invention.

[0260] Figure 2A The methods include:

[0261] The target organ was irradiated with high-level ultrasound during the first time period (202);

[0262] During the second time period, the target organ was irradiated with lower levels of ultrasound (204); and

[0263] Cell samples were collected (206).

[0264] In some implementations, there is an optional time period (not shown) between the first time period of higher-level ultrasound irradiation and the second time period of lower-level ultrasound irradiation, during which no ultrasound irradiation occurs.

[0265] Now for reference Figure 2B The figure is a simplified flowchart of a method for generating cytopathological cell samples according to an exemplary embodiment of the present invention.

[0266] Figure 2B The methods include:

[0267] The target organ was irradiated with high-level ultrasound during the first time period (222);

[0268] During the second time period, the target organ was irradiated with lower levels of ultrasound (214); and

[0269] Cell samples were collected (216).

[0270] In some implementations, there is an optional time period (not shown) between a first time period corresponding to a lower level of ultrasound irradiation and a second time period corresponding to a higher level of ultrasound irradiation, during which no ultrasound irradiation occurs.

[0271] Now for reference Figure 2C , Figure 2C This is a simplified flowchart of a method for generating cytopathological cell samples according to an exemplary embodiment of the present invention.

[0272] Figure 2C The methods include:

[0273] During the first time period, the target tissue was irradiated with ultrasound energy of the first intensity level (222);

[0274] During the second time period, the target tissue was irradiated with ultrasound at a second intensity level (224);

[0275] in

[0276] The first intensity level is higher than the second intensity level.

[0277] Now for reference Figure 2D The figure is a simplified flowchart of a method for inducing exfoliation according to an exemplary embodiment of the present invention.

[0278] Figure 2D The method includes irradiating the target tissue with ultrasound energy corresponding to a mechanical index (MI) in the range of 0.3 to 1.8 (242).

[0279] In some implementations, the target tissue is irradiated with a focused ultrasound beam.

[0280] In some implementations, ultrasonic irradiation is performed by repeating ultrasonic pulses with a duration ranging from 20 microseconds to 800 microseconds.

[0281] In some implementations, ultrasonic irradiation is performed by repeating ultrasonic pulses at a rate of 1 to 1400 times per second.

[0282] Now for reference Figures 3A-3E These are simplified illustrations of the use of ultrasound and ultrasound contrast agents to generate cell samples according to exemplary embodiments of the present invention.

[0283] Figure 3A An example of a three-layered tissue is shown. The target organ consists of a first layer of epithelium 302, a second layer of basement membrane (lamina propria) 303, and a third layer of submucosal layer 304, which are attached to each other horizontally and vertically. Note the presence of capillaries in the third layer of submucosal layer 304.

[0284] Figure 3B The image shows three cell layers 302, 303, and 304 after ultrasound irradiation, in which a group of epithelial cells 306 has separated from the other cells.

[0285] In some implementations, separation is achieved by inducing shear waves through ultrasonic irradiation.

[0286] In some implementations, the group of cells 306 can be completely isolated and can flow along the pancreatic duct and be collected as a cell sample.

[0287] In some implementations, the separation of the group of cells 306 may optionally be accomplished using additional ultrasound irradiation at an energy level lower than that used to generate the shear wave.

[0288] In some implementations, an ultrasound contrast agent material, such as secretin, is provided to the target organ, and optionally the target organ is further irradiated with ultrasound at a lower energy level than that used to generate the shear wave.

[0289] Figure 3C The diagram shows three cell layers 302, 303, and 304, the group of cells 306, and bubbles 308 formed in the capillaries of the third submucosal layer 304 by ultrasound irradiation with an ultrasound contrast agent. These bubbles further serve to separate the group of cells 306 from their adjacent cells.

[0290] Figure 3D The group of cells 306 is shown separated from the adjacent cells, which potentially allows the group of cells 306 to flow with the surrounding fluid and optionally be collected in a cell sample.

[0291] As a non-limiting example, a cell layer 302 may be an epithelial layer in the pancreatic duct, and the group of cells 306 may be a group of epithelial cells isolated from pancreatic tissue to flow along the pancreatic duct and optionally be collected as a cell sample.

[0292] Now for reference Figure 3E This is a simplified illustration of generating a cell sample using ultrasound and an ultrasound contrast agent according to an exemplary embodiment of the present invention.

[0293] Figure 3E Epithelial cells 312 and 316, basement membrane 314, and blood vessel 317 irradiated by ultrasound 319 are shown.

[0294] Described based on the pancreas Figure 3E , Figure 3E The pancreatic ductal epithelium 312, 316 is visualized as a single layer of columnar cells 312, 315, 316 atop the basement membrane 314. Shear waves generated by ultrasound irradiation disrupt the transverse bonds between the columnar cells 312, 315, 316, and subsequent wide-beam ultrasound irradiation 319 from microvesicles 318 disrupts the bond between the basal bands of the columnar cells 315, 316 and the underlying basement membrane 314. Ultrasound irradiation 319 does not disrupt the basal bonds between one layer of glandular cells and the layer of glandular cells beneath them.

[0295] Various example implementation schemes

[0296] In some embodiments, the ultrasound energy absorbed within the target tissue, as a non-limiting example within the pancreas, causes mechanical vibrations that enhance the separation and / or detachment of pancreatic ductal cells and, optionally, tissue debris. In the case of obtaining a pancreatic sample, secretin (a substance that induces pancreatic secretion) is then optionally injected. Some of the detached and exfoliated cells and tissue debris may deposit in pancreatic juice, which is then collected, for example, via endoscopy.

[0297] In some implementations, the cells and / or tissue fragments in the obtained cell-rich sample are then subjected to morphological analysis and / or molecular biomarkers to detect the presence or absence of cellular abnormalities.

[0298] Non-limiting example methods for obtaining cell samples and tissue fragments from a subject's organ may include: irradiating the organ with a narrow-beam high-intensity ultrasound energy that effectively induces shock waves and / or shear waves, thereby causing relevant vibrations in the tissue or organ; subsequently irradiating the subject's organ with a wide-beam, low-intensity ultrasound with a longer pulse width, while simultaneously injecting the subject with an ultrasound contrast agent, thereby causing cell or epithelial tissue fragments to detach from the subject's organ.

[0299] These methods may increase the total number of cells expressed in pancreatic juice. Furthermore, the procedures described herein can induce the separation of intact tissue fragments from the pancreas.

[0300] When applied to other organs or body parts such as the mediastinum, pleura, pericardium, peritoneum, lungs, breast, salivary glands, meninges, pancreatic duct, pancreatic cysts, kidneys, liver, prostate, bladder, or ovaries, these methods potentially increase the total number of cells expressed in surrounding fluids (e.g., fluids found in and / or near the organ and / or drained into the organ). Furthermore, the methods described herein potentially induce the separation of intact tissue fragments from the aforementioned exemplary organs. For example, the methods described herein can be used to induce lung cells to detach into surrounding sputum, or to induce bladder cells and prostate and / or bladder tissue to detach into surrounding urine.

[0301] In some implementations, a method is provided for obtaining cell samples and tissue fragments from a subject's organ, comprising: irradiating a target organ of the subject with an amount of ultrasonic energy, said ultrasonic energy effectively inducing shear waves that potentially induce relevant vibrations in the target tissue or organ.

[0302] In some embodiments, a method for obtaining cell samples and tissue fragments from an organ of a subject is provided, comprising: irradiating a target organ of the subject with ultrasound energy of a certain amount, said ultrasound energy effectively causing shear waves that potentially cause relevant vibrations in the target tissue or organ; and subsequently irradiating the target organ of the subject with wide-beam, low-intensity, long-pulse ultrasound after the subject has been injected with an ultrasound contrast agent, thereby causing cell or epithelial tissue fragments to detach from the target organ.

[0303] In some implementations, a method for obtaining pancreatic cells from a subject is provided, comprising: irradiating the subject's pancreas with a certain amount of single-frequency high-energy pulsed ultrasound for a duration of 2-40 microseconds, effectively inducing cell detachment into the subject's pancreatic duct.

[0304] In some implementations, the ultrasound is emitted at a frequency of 3 MHz.

[0305] In some implementations, ultrasound is emitted at different frequencies within a range associated with ultrasound.

[0306] In some implementations, a method for obtaining pancreatic cells from a subject is provided, comprising: irradiating the subject's pancreas with an amount of ultrasound energy from a multi-frequency array, effectively obtaining asymmetric ultrasound waves at predetermined points in the pancreas, either mostly positive or mostly negative, followed by mostly positive, and inducing cell detachment into the subject's pancreatic ducts. "Mostly positive" and "mostly negative" refer to pulses obtained through the constructive interference of two or more ultrasound waves, optionally at integer multiples of their fundamental transmission frequencies, such as fundamental transmission frequencies f, 2f, 3f, etc., while the phase between the transmission frequencies is designed to produce the sum of all positive (or negative) amplitudes at a specific depth within the tissue.

[0307] In some implementation schemes, methods for treating pancreatic diseases in subjects are provided, including:

[0308] a) To determine whether the subject's pancreas contains abnormally developing pancreatic cells or pancreatic cancer cells using the methods described herein; and

[0309] b) Subjects found to have abnormal pancreatic cells or pancreatic cancer cells in their pancreas in a) undergo chemotherapy, radiotherapy, immunotherapy, or pancreatectomy.

[0310] In some implementations, one or more of cell morphology analysis, tissue morphology analysis, or molecular marker analysis are used to determine whether the subject's pancreas contains abnormal pancreatic cells or pancreatic cancer cells.

[0311] In some embodiments, a method is provided for increasing the efficiency of collecting cell samples from tissues or organs during a measurement of a subject, comprising irradiating the subject's tissues or organs with an amount of ultrasound energy prior to collecting the cell samples, said ultrasound energy effectively inducing shear waves within the tissues or organs, thereby inducing cell or tissue fragments to detach from the tissues or organs, followed by collection of the cell samples.

[0312] In some embodiments, a method is provided for measuring a cell or tissue sample from a subject to determine whether the cells or tissue contain cancerous or precancerous cells or tissue, comprising:

[0313] a) Receive a sample of cells or tissue, wherein the sample has been obtained in advance by generating shear waves in the subject's tissues and organs to induce the shedding of cell or epithelial tissue fragments from the subject's organs, and collect the sample of the shed cells or tissue.

[0314] b) Perform one or more of the following: cell morphology analysis, tissue morphology analysis, and molecular marker analysis to determine whether the cells or tissues contain cancerous or precancerous cells or tissues.

[0315] In some implementations, a method for early detection of abnormally developing cells and cancer cells in the pancreas is provided, comprising applying ultrasound energy to the pancreas to induce cell shedding, followed by endoscopic collection of pancreatic juice containing shed cells and / or cell clusters for molecular and microscopic morphological examination.

[0316] In some embodiments, a bodily fluid sample is provided, wherein the sample has been obtained directly from a subject whose tissue or organ has been irradiated with ultrasound, and wherein the sample contains epithelial cells or other cells from the tissue or organ, and the content of such epithelial cells or other cells is enriched / increased by more than 2 times relative to the content of epithelial cells or other cells in other identical samples obtained from a subject whose tissue or organ has not been irradiated with ultrasound.

[0317] In some implementations, a method for obtaining serial pancreatic duct cell samples from a subject is provided, comprising:

[0318] Before or within one hour after the start of ultrasound energy irradiation, a certain amount of secretin peptide is administered to the subject to effectively induce pancreatic secretion.

[0319] The subject's pancreas was irradiated with a predetermined amount of ultrasound energy at a designated point in the pancreas to induce cell detachment and entry into the pancreatic duct; and

[0320] A sample containing fluid from adjacent pancreatic duct cells was taken from the subject.

[0321] In some embodiments of the present invention, the methods and / or systems described herein are used to induce prostate cell shedding.

[0322] Now for reference Figure 4A The figure is a front perspective view of an ultrasonic probe according to an embodiment of the present invention.

[0323] Figure 4A A front view of an ultrasound transducer or probe 410 is shown. The probe 410 is formed by a housing and one or more piezoelectric elements housed within the housing 412. In some embodiments of the invention, the housing 412 has a strip 414 attached thereto for attaching the probe to a patient.

[0324] Now for reference Figure 4B , Figure 4B This is a schematic diagram of a probe configuration according to an exemplary embodiment of the present invention.

[0325] Figure 4B It shows Figure 4A The housing 410, and the arrangement of one or more ultrasonic piezoelectric elements 422 within the housing 410.

[0326] In some embodiments, one or more optional receiving elements 424 are optionally arranged within the housing 410. In some embodiments, the receiving element 424 is optionally a cavitation detector.

[0327] In some implementations, probe 412 may optionally be programmed to generate a non-focused ultrasonic energy beam.

[0328] In some implementations, one or more ultrasonic transducers 422 in probe 412 may optionally be programmed to generate a focused ultrasonic energy beam.

[0329] In some implementations, multiple ultrasonic transducers 422 in probe 412 may optionally be programmed to generate a focused ultrasonic energy beam together.

[0330] Those skilled in the art will understand that any number of piezoelectric elements can be used in different embodiments of the present invention.

[0331] In some embodiments of the invention, one or more cavitation detectors 424 may be optionally disposed on the probe to detect the occurrence of unstable cavitation. The cavitation detector 424 may include a passive cavitation detector (PCD) or another type of hydrophone.

[0332] Now for reference Figure 4C , Figure 4C This is a simplified diagram of an array of elements in an ultrasonic transducer according to an exemplary embodiment of the present invention.

[0333] Figure 4CA one-dimensional array 431 is intended to illustrate the elongated transducer unit 432.

[0334] In some embodiments, the ultrasonic probe or transducer comprises multiple transducer crystals, or is formed from a single crystal processed by etching or a similar dicing process to form a uniformly arranged series of thin / fine transducer units 432. The transducer units may be arranged on a flat, curved, or concave substrate.

[0335] In some embodiments, the probe is configured to deliver a non-focused uniform beam, or a controlled spot-focused or controlled strip-focused beam, to a target organ such as the pancreas with sufficient radiation intensity to impart sufficient energy to the tissue of the target organ, and in some embodiments, to impart sufficient energy to the contrast agent flowing within its vascular system to induce cell and / or tissue ablation.

[0336] In some embodiments, the transducer comprises an array 431 of crystal elements 432, which are elongated, approximately 120 mm in length (as a non-limiting example), and narrow, approximately 4 mm in width (as a non-limiting example). Figure 4C As shown. Exciting a subset of these elements can produce a strip-shaped, controllable beam with approximate dimensions of 3 mm × 10 mm on the focal plane.

[0337] In some embodiments, the number of transducer crystal elements 431 can range from 2 to 10, or even up to 2 to 100. In some embodiments, the number of transducer crystal elements 431 is odd, enabling a symmetrical distribution of the central transducer crystal element and the transducer crystal element partitions on either side of the central transducer crystal element.

[0338] Now for reference Figure 4D The figure is a simplified diagram of an array of elements in an ultrasonic transducer according to an exemplary embodiment of the present invention.

[0339] Figure 4D The purpose is to illustrate a two-dimensional array 435 of transducer unit 436.

[0340] In some implementation schemes, such as Figure 4C The transducer crystal element 431 shown is etched into multiple sections along the vertical axis, such as... Figure 4D As shown. As a non-limiting example, the number of partitions can range from 2 to 10, or even as high as 2 to 100. In some implementations, the number of partitions is odd, making it possible to define a central partition and a symmetrical distribution of partitions on either side of the central partition.

[0341] Excitation of this element or a subset of this element can produce a point-focused, controllable beam.

[0342] In some implementations, the focal point of the beam may have a size of approximately 2 mm × 4 mm on the focal plane.

[0343] In several embodiments, the ultrasound probe is configured to sequentially or simultaneously irradiate the following combinations of ultrasound: 1) long pulses of a point-focused or strip-focused controllable beam; 2) long pulses of a substantially uniform, flat beam that substantially simultaneously covers the entire area (e.g., an organ or organ region).

[0344] In some implementations, the probe (a 2D array of transducer crystals, or a single crystal etched into multiple columns and rows) enables sequential or simultaneous ultrasonic irradiation of a combination of long pulses of point-focused or strip-controlled beams and long pulses of substantially uniform flat beams, and also allows for 3D imaging using lateral and longitudinal focusing.

[0345] In some implementations, compared to larger transducer crystals and / or unetched large transducer crystals, probes (2D arrays of transducer crystals, or individual crystals etched into multiple columns and / or rows) can achieve reduced peak excitation intensity and better power management per crystal element, as well as probe thermal control.

[0346] In some implementations, the probe includes multiple cavitation detectors and an associated computer system configured to: i) detect the occurrence of inertial cavitation; ii) determine the plane in which inertial cavitation is detected; iii) determine whether the plane in which such cavitation is detected is located within the target organ; and iv) reduce the intensity of ultrasound energy if the plane in which inertial cavitation is detected is within the plane identified as being within the target organ.

[0347] In some implementations, a system is provided that includes a probe cable and connector, a tape, an electronic module, a control unit, a processing unit, and a graphical user interface (GUI).

[0348] In some implementations, the system may optionally include a disposable sheath that encapsulates the probe and / or its cable, or encapsulates at least a portion of the probe and / or cable, thereby allowing aseptic use of the system while allowing attachment to a strap that holds the probe in place.

[0349] Now for reference Figure 6 , Figure 6This is a flowchart of a method for generating cytopathological cell samples from the prostate gland according to an exemplary embodiment of the present invention. In some embodiments, the methods and / or systems described herein are used to induce the shedding of prostate cells and / or tissue, with necessary modifications. The shed cells and / or tissue are expelled into the urethra and expelled from the body and / or into the bladder during ejaculation. These cells are then expelled with the subject's semen and / or urine and collected for analysis, potentially allowing for early detection of prostate cancer and precancerous conditions. The collection of urine and / or semen is a non-invasive and / or minimally invasive procedure, with the potential advantage of reducing patient discomfort and / or pain and increasing prostate cancer testing compliance. It should be understood that, in this embodiment of the invention, the step of administering secretin / secretin, etc., may optionally be omitted.

[0350] Figure 6 The methods include:

[0351] Optionally, patients (602) are selected. Prostate cancer is the most common cancer in the United States, and if not detected early, it can be debilitating or fatal. However, early detection can potentially increase the chances of successful treatment. Because prostate cancer often has no symptoms before the disease progresses / becomes advanced, this method can be used for prostate cancer screening. As an alternative to or supplement to prostate cancer screening, this method can be used for prostate cancer surveillance. For example, monitoring patients diagnosed with prostate cancer, and optionally without aggressive treatment. Another example is monitoring patients with previous results that are abnormal in the collected samples.

[0352] Place an ultrasound probe (e.g., an ultrasound transducer) (604), optionally placing it near the prostate, for example, as... Figure 7 As shown. Proximity to the prostate has the potential advantage of reducing and / or avoiding damage to surrounding blood vessels and / or nerves, bladder damage, and / or causing erectile dysfunction. In some embodiments, the probe is introduced into a predetermined point near the prostate, optionally transrectally.

[0353] A predetermined point in the prostate gland is irradiated with a certain amount of ultrasound energy (606) to induce cell shedding, which optionally allows the shed cells to enter the subject's urethra. In some embodiments, the ultrasound energy corresponds to a higher intensity for a period of time.

[0354] As described above, in some embodiments of the invention, the shear wave is generated by a focused ultrasound beam pulse of high intensity and long duration (e.g., 100 microseconds to 800 microseconds). For example, pulse durations of about 100 microseconds to 800 microseconds, or 50 microseconds to 100 microseconds, or 150 microseconds to 300 microseconds, or about 200 microseconds, or about 250 microseconds, or about 180 microseconds, or lower or higher, or intermediate microseconds. Typically, this ultrasound beam is perpendicular to the surface of the prostate epithelium. For example, the ultrasound beam is at an angle of about 60 degrees to 120 degrees relative to the rectum and / or perineum, or in the range of about 75 degrees to 105 degrees, or in the range of about 80 degrees to 100 degrees, or about 85 degrees, or about 95 degrees, or lower or higher, or intermediate degrees. The ultrasound beam that generates the shear wave passes through the prostate and produces pressure changes that cause rapid expansion and contraction, generally parallel to the surface plane of the tissue. These mechanical changes potentially cause dissociation in the lateral connections between epithelial cells. Note that standard imaging ultrasound uses a pulse width of 2 microseconds. In some embodiments of the invention, ultrasound irradiation is performed with higher intensity ultrasound energy during a first time period, optionally followed by ultrasound irradiation with lower intensity ultrasound energy during a second time period. An idle time period may or may not exist between the ultrasound irradiation of the first time period and the ultrasound irradiation of the second time period. The second time period potentially lifts part of the segmented epithelium from the organ surface, thereby inducing the shedding of cells and tissue debris.

[0355] In some examples, the first time period of ultrasound irradiation lasts for 1 to 15 minutes with a higher MI shear wave. For example, in the range of 0.5 to 11 minutes, in the range of 10 to 20 minutes, in the range of 5 to 30 minutes, or about 5 minutes, or about 12 minutes, or about 17 minutes, or lower, higher, or intermediate minutes. In some examples, the second time period of ultrasound irradiation lasts for 10 to 20 minutes. In some examples, the second time period of ultrasound irradiation lasts for 3 to 30 minutes. For example, in the range of 1 to 15 minutes, in the range of 8 to 35 minutes, in the range of 15 to 45 minutes, or about 9 minutes, or about 20 minutes, or about 25 minutes, or lower, higher, or intermediate minutes.

[0356] Alternatively or additionally, the prostate may be irradiated with lower-intensity ultrasound before the application of higher-intensity ultrasound. In several other embodiments, the patient's prostate may be irradiated with ultrasound according to other methods, for example, as described in patent applications US17 / 367,658 and / or US17 / 028,588. In some embodiments, this method may be disadvantageous for treating the prostate because the relatively increased number of microbubbles and / or prolonged duration of microbubble ultrasound irradiation may lead to undesirable interactions with the prostate's surrounding environment, such as blood cells, nerves, and organs, such as the bladder and urethra.

[0357] Optionally, an ultrasound contrast agent (908) is administered to the patient. In some embodiments, the contrast agent is administered after a first period of ultrasound irradiation (e.g., with high-intensity ultrasound irradiation). The contrast agent may be gas microbubbles of 1-4 μm in size, optionally encapsulated in a protein, lipid, or polymer shell, which expand and contract due to changes in their local pressure when exposed to ultrasound. Thus, when intravenously injected into the subject, the microbubbles flow through the vascular system and all its capillaries, reaching the rich microvascular system surrounding the prostate epithelial cells. In some embodiments, the combined action of the ultrasound beam aimed at the prostate and the presence of microbubbles within its microvascular system leads to the shedding of cells and / or tissue fragments. Unbound from theory, this may result in the synchronous expansion-contraction of the microvascular system beneath the epithelial layer, thereby dislocating cells and cell clusters by disrupting the cellular connections between the basement membrane of the epithelial cells and the underlying basement membrane. In some embodiments, the prostate is subjected to wide-area ultrasound energy after the introduction of microbubbles. The ultrasound application of embodiments of the present invention can be described as low-intensity unfocused ultrasound (LINFU application®). In some implementations, the first period of ultrasound irradiation, which optionally generates shear waves at a higher MI, lasts from 1 minute to 15 minutes, optionally about 5 minutes. For example, it can range from 0.5 minutes to 11 minutes, from 10 minutes to 20 minutes, from 5 minutes to 30 minutes, or about 5 minutes, or about 12 minutes, or about 17 minutes, or a lower, higher, or intermediate number of minutes. Following the first period is a second period of microbubble ultrasound irradiation at a lower MI, lasting approximately 10 minutes to 20 minutes. For example, it can range from 5 minutes to 12 minutes, from 8 minutes to 25 minutes, from 15 minutes to 30 minutes, or about 6 minutes, or about 22 minutes, or about 25 minutes, or a lower, higher, or intermediate number of minutes.

[0358] Optionally, prostatic fluid secretion is promoted (910). Prostatic fluid secretion potentially allows for relatively rapid acquisition of exfoliated cells and / or tissue fragments shed from the prostate, with the potential advantage of reducing the time and / or volume of sample required for collection (e.g., prostatic fluid volume). In some embodiments, the promotion of secretion includes promoting ejaculation, for example, through sexual arousal. In some embodiments, the promotion of secretion includes stimulating the prostate, with the potential advantage of allowing the promotion of prostatic fluid secretion in patients suffering from erectile dysfunction and / or unable to ejaculate due to other causes (e.g., spinal cord injury or other neurological disorders).

[0359] In some embodiments, the stimulation includes electroejaculation, which involves sending an electric current to the prostate. Optionally, the current causes the prostate and seminal vesicles to contract and release prostatic secretions (e.g., semen). Alternatively or additionally, the stimulation includes, for example, pressing the prostate during a rectal examination and / or prostate massage.

[0360] Optionally, urination is encouraged (912) because exfoliated epithelial cells and / or tissue can be drained into the bladder and subsequently expelled with urine. Encouraging urination has the potential advantage of allowing relatively rapid acquisition of exfoliated cells and / or tissue fragments from the prostate, and the potential advantage of reducing the time and / or volume of sample required for collection (e.g., urine volume). In some embodiments, inducing urination includes administering at least one diuretic substance, such as furosemide, to the patient. Alternatively or additionally, the patient is prompted to consume fluids before, after, and / or during ultrasound treatment. Optionally, the prostate is irradiated with ultrasound when the patient's bladder is full and / or nearly full.

[0361] In some implementations, sterile urine samples are obtained during and / or after ultrasound irradiation of the prostate via catheterization (e.g., connecting a catheter to the patient's bladder for urine collection). Catheterization has the potential advantage of reducing and / or avoiding sample loss and / or contamination during patient urine collection.

[0362] Collect a sample (904). The sample includes exfoliated cells and / or tissue fragments from one or both of collected urine and collected semen (e.g., prostatic secretions). The sample contains at least one of the following: cells exfoliated from the prostate, adjacent / neighboring cells, cell sheets, cell clusters, and / or cell clumps; and / or at least one tissue fragment exfoliated from the prostate. In some embodiments of the invention, cells (e.g., any one of cells, cell clusters / clumps, cell sheets, and / or tissue fragments) are exfoliated from the prostate in greater quantities compared to a prostate that has not been irradiated with ultrasound and / or a prostate that has not been irradiated with relatively high-intensity focused ultrasound to generate shear waves. In some embodiments, samples are obtained from the prostate at a greater rate (i.e., the number of samples per unit time) compared to a prostate that has not been irradiated with ultrasound and / or a prostate that has not been irradiated with relatively high-intensity focused ultrasound to generate shear waves. In some embodiments of the invention, the method is used to obtain cell sheets, and / or cell populations, and / or tissue fragment samples containing a greater number of cells than a prostate that has not been irradiated with ultrasound and / or a prostate that has not been irradiated with relatively high-intensity focused ultrasound to generate shear waves; and

[0363] The collected sample (916) is tested. In some embodiments, the test includes processing the sample to isolate epithelial cells and / or concentrate epithelial cells in the sample (e.g., by using a cell centrifuge, also known as a "cell centrifuge smear"). In embodiments of the invention, the test includes, optionally, analysis of the sample by a pathologist. In some embodiments, the test includes cytopathological examination of cells and / or tissue fragments from the sample.

[0364] In some implementations, cell morphology analysis, tissue morphology analysis, molecular biological analysis, and molecular biomarkers are used to detect the presence or absence of cellular abnormalities (e.g., identifying whether cells are cancerous or developmentally abnormal). In some implementations, tissue morphology analysis is performed. Evaluation of tissue fragments (optionally, intact tissue fragments) allows for the assessment of the contrast between the honeycomb pattern of cells characteristic of normal epithelium and the loss of honeycomb pattern occurring in developmental abnormalities and cancer. This evaluation potentially allows for the differentiation of atypical cells from inflammatory and developmentally abnormal cells.

[0365] In some embodiments, the method includes irradiating the entire prostate and / or substantially the entire prostate with ultrasound in a single-stage process. Alternatively or additionally, the method includes irradiating the prostate with ultrasound in more than one step, wherein each step includes irradiating a portion and / or a region of the prostate with ultrasound and collecting a sample from there. This stepwise procedure potentially allows for the detection of the location of abnormally developing cells or pancreatic cancer cells within the prostate. In some embodiments, distributed ultrasound irradiation of the prostate is achieved by adjusting the positioning of the ultrasound probe and / or adjusting the ultrasound focus to irradiate different regions of the prostate with ultrasound at each stage. In some embodiments, the ultrasound probe (e.g., Figure 8 The probes 802, 804, and / or 806 described herein include multiple independent transducers, each of which can be aimed at and / or focused on different parts of the prostate. In some embodiments, the probes include high-intensity focused ultrasound.

[0366] Now for reference Figure 7 A schematic cross-sectional view of a prostatic sac 440 and a segment of a prostatic duct (which ultimately drains into the urethra) according to an exemplary embodiment of the present invention is shown. As shown, an epithelial cell layer 442 (including basal cells, luminal cells, and neuroendocrine cells) is located on top of a basement membrane 444. In embodiments of the invention, a method is provided for draining epithelial cells (e.g., cell 448), epithelial cell clusters (e.g., cell cluster 450), and / or epithelial tissue fragments, which are subsequently drained in ejaculation and / or into the bladder, and subsequently in urine. In embodiments of the invention, the method includes the step of irradiating the prostate of a subject with an ultrasound probe having a certain amount of ultrasound energy at predetermined points in the prostate to induce cell detachment into the segment of the prostatic duct 446 and ultimately into the subject's urethra.

[0367] Now for reference Figure 8 A schematic cross-sectional view of a patient's pelvic region and a simplified diagram of an ultrasound probe interacting with the patient's prostate are shown, according to an exemplary embodiment of the present invention.

[0368] In some implementations, the ultrasound device is configured and / or programmed to employ methods for producing cytopathological cell samples from the prostate (as described herein, for example in...). Figure 6 (in the middle), which includes an ultrasound probe configured for ultrasound irradiation of the prostate. In some embodiments, the probe is a variant of probe 410, adjusted for the prostate, for example, its size and / or shape adapted to the pelvic region.

[0369] An ultrasonic probe includes a transducer, a power supply, and a controller that uses a power supply from the power source to operate the transducer. In some implementations, the transducer is configured to transmit a focused, controllable beam.

[0370] In some embodiments, the probe includes a transrectal ultrasound probe 802. The probe is configured to be inserted into the patient's rectum and reach the prostate. In some embodiments, the transrectal ultrasound probe 802 includes a concave contact surface 801 conforming to the shape of the prostate surface. In some embodiments, the transrectal ultrasound probe 802 potentially allows ultrasound irradiation of the entire prostate and / or substantially the entire prostate. Alternatively or additionally, the transrectal ultrasound probe 802 is configured to irradiate a portion / region of the prostate; for example, in some embodiments, the contact surface of the probe is divided into multiple regions, each region for irradiating a different portion of the prostate. In some embodiments, the probe includes multiple independent transducers, potentially allowing testing of only a portion of the prostate when detecting an abnormality in said portion. Testing the prostate by testing only a portion of the prostate at a time potentially allows detection of the location of abnormalities within the prostate.

[0371] In some implementations, the ultrasound device includes a transperineal ultrasound probe 804. The transperineal ultrasound probe 804 is configured to be positioned on the perineum and is intended to deliver ultrasound waves to the prostate.

[0372] Applying ultrasound externally (e.g., probe 804 contacting the perineum) has the potential advantage of reducing patient discomfort and / or pain.

[0373] In some embodiments, the ultrasound device includes an ultrasound catheter probe 806 configured for insertion into a patient's urethra. The probe 806 includes an ultrasound transducer located at its distal end, which is connected to a flexible, elongated shaft, allowing insertion into and access to the prostate via the urethra. In some embodiments, the transducer at the distal end of the probe 806 is used to irradiate the prostate with ultrasound in more than one phase. In each phase, another portion (e.g., a region of the prostate) is irradiated with ultrasound, and at least one sample is collected therefrom. If abnormal cells are detected, multiple phases allow for identification of the location of the abnormal cells. In some embodiments, the probe 806 includes more than one transducer at its distal end and / or along its axis, allowing for ultrasound irradiation of a wider portion and / or the entire and / or substantially the entire prostate (e.g., in a single phase).

[0374] In some embodiments, the probes (e.g., 802, 804, and / or 806) include an imaging system for monitoring the position and / or orientation of the probes. In some embodiments, the imaging system is located at the tip of the probe (e.g., the tip 801 of probe 802, the tip of probe 806, and / or the contact surface of probe 804), and / or near the transducer of the probe. In some embodiments, the imaging system includes an ultrasound imaging system. In some embodiments, in addition to ultrasound irradiation, the transducer of the probe is also used for imaging, optionally, imaging is performed between ultrasound irradiation pulses. This monitoring potentially ensures that the probe is aligned with the prostate and / or focused within the prostate. In some embodiments, the imaging system is connected to a controller that warns and / or stops applying ultrasound if deviation from the prostate is detected. For example, if the patient and / or physician moves, or due to displacement of internal organs, such as a full bladder. Monitoring the orientation of the probe has the potential advantage of reducing and / or avoiding the risk of damage to nerves, blood vessels, and / or organs (such as the bladder) surrounding the prostate due to incorrect probe orientation. In some implementations, the imaging system includes a controller for identifying the prostate, which may optionally be initially tagged by a human operator and / or an AI (artificial intelligence) component.

[0375] In some embodiments, the ultrasound probe (e.g., probes 802, 804, and / or 806) includes an electroejaculation device / component. In some embodiments, the probe includes electrodes for delivering electrical stimulation and a controller for operating and adjusting the intensity and frequency of the electrical pulses.

[0376] In some embodiments, the ultrasound probe (e.g., probes 802, 804, and / or 806) includes a prostate stimulator (e.g., a prostate massager) configured to apply gentle pressure and / or vibration to the prostate to promote prostate secretion. For example, probe 802 includes the stimulator at its distal end.

[0377] It is anticipated that many related ultrasonic transducers will be developed during the term of this patent application, and the scope of the term "ultrasound" is intended to a priori include the ultrasound produced by all these new technologies.

[0378] It is anticipated that many related ultrasound contrast materials will be developed during the term of this patent application, and the scope of the term "ultrasound contrast materials" is intended to include all of these new ultrasound contrast materials a priori.

[0379] As used in this article, the terms “approximately” and “about” in relation to quantity or value mean “within ±25% of”.

[0380] The terms “including,” “comprising,” “having,” and their variations mean “including but not limited to.”

[0381] The term “composed of / consisting of” is intended to mean “including and limited to”.

[0382] The term "consistent with / made up of" means that a composition, method, or structure may include additional ingredients, steps, and / or portions, provided that the additional ingredients, steps, and / or portions do not materially alter the essential and novel features of the claimed composition, method, or structure.

[0383] As used herein, the singular forms “a,” “an,” and “the” include the plural unless the context clearly indicates otherwise. For example, the terms “a unit” or “at least one unit” can include multiple units, including combinations thereof.

[0384] The terms “example” and “exemplary” as used herein mean “used as an example, instance, or illustration.” Any implementation described as “example” or “exemplary” is not necessarily to be construed as superior to or advantageous to other implementations, and / or as excluding the combination of features with other implementations.

[0385] The term “optionally” is used herein to mean “provided in some embodiments but not in others”. Any particular embodiment of this disclosure may include several “optional” features unless these features conflict with each other.

[0386] Throughout this application, various embodiments of this disclosure may be presented in the form of ranges. It should be understood that the range format is for convenience and brevity only and should not be construed as an immutable limitation on the scope of this disclosure. Therefore, the description of a range should be considered as having specifically disclosed all possible subranges and individual numerical values ​​within that range. For example, a description of a range such as 1 to 6 should be considered as having specifically disclosed subranges such as 1 to 3, 1 to 4, 1 to 5, 2 to 4, 2 to 6, 3 to 6, etc., and individual numbers within that range, such as 1, 2, 3, 4, 5, and 6. This applies to any range width.

[0387] Whenever a range of numbers is indicated herein (e.g., “10–15”, “10 to 15”, or any pair of numbers connected by such another range indication), it is intended to include any number (fraction or integer) within the indicated range boundaries, including the range boundaries, unless the context explicitly states otherwise. The phrases “range between the first and second indicators” and “range from the first indicator to / to the second indicator” (or another such range indication term) are used interchangeably herein and mean to include the first and second indicators and all fractions and integers in between.

[0388] Unless otherwise stated, the numerical values ​​used herein and any ranges of numerical values ​​based thereon are approximations within the accuracy of reasonable measurement and rounding errors as understood by those skilled in the art.

[0389] As used herein, the term “method” refers to the manner, means, techniques and procedures used to accomplish a given task, including but not limited to those manner, means, techniques and procedures known to practitioners in the fields of chemistry, pharmacology, biology, biochemistry and medicine, or readily developed from known manner, means, techniques and procedures.

[0390] As used in this article, the term “treatment” includes eliminating, substantially inhibiting, slowing or reversing the progression of a disease, substantially improving the clinical or aesthetic symptoms of a disease, or substantially preventing the occurrence of the clinical or aesthetic symptoms of a disease.

[0391] It should be understood that, for clarity, certain features of this disclosure described in the context of individual embodiments may also be provided in combination in a single embodiment. Conversely, for brevity, various features of this disclosure described in the context of individual embodiments may also be provided individually, or in any suitable sub-combination, or appropriately provided in any other described embodiments of this disclosure. Certain features described in the context of various embodiments should not be considered essential features of those embodiments unless the embodiment would not function without these elements.

[0392] As described above and as claimed in the following claims portion, various embodiments and aspects of this disclosure find experimental support in the following embodiments.

[0393] Example

[0394] The following embodiments, together with the foregoing description, illustrate some implementations of this disclosure in a non-limiting manner.

[0395] Now for reference Figure 5 , Figure 5 This is an image of a group of adjacent cells obtained through an exemplary embodiment of the present invention.

[0396] Figure 5 Group 402 or sheet 402 of pancreatic duct cells are shown.

[0397] Experimental results

[0398] Now for reference Figure 9 , Figure 9 This is a table of experimental results from a clinical registry (at Sarasota Memorial Hospital) based on an exemplary embodiment of the present invention.

[0399] Twelve patients underwent ultrasound irradiation. According to an embodiment of the invention described herein, the ultrasound irradiation protocol involved irradiating the pancreas with high-intensity, partially focused ultrasound waves without microbubble contrast agents for a period of time to generate shear waves, followed by ultrasound irradiation with lower-intensity, lower-focus ultrasound waves combined with microbubbles. Pancreatic cell samples collected from all 12 patients contained sufficient cells for the results to “meet analytical requirements.” Atypical cells were detected in three patients (patients 1, 5, and 12), which altered the treatment course for these patients. Patient 1 had previously undergone five years of monitoring. Based on the surgical outcome, he was recommended for partial pancreatectomy. Patient 5 had previously undergone five years of monitoring. Due to the surgical outcome, she was recommended for monitoring every six months. Patient 12 was scheduled to review the surgical outcome with her internist and surgeon and might be recommended for partial pancreatectomy. The other patients were asked to repeat this procedure within one year.

[0400] Although the invention has been described in conjunction with specific embodiments thereof, it will be apparent to those skilled in the art that many substitutions, modifications, and variations will be readily apparent. Therefore, the invention is intended to encompass all such substitutions, modifications, and variations falling within the spirit and broad scope of the appended claims.

[0401] The applicant intends that all publications, patents, and patent applications mentioned in this specification are incorporated herein by reference in their entirety, and when each individual publication, patent, or patent application is mentioned herein by reference in its entirety, it is as if each individual publication, patent, or patent application were specifically and individually cited in the reference. Furthermore, any reference or designation of any reference in this application should not be construed as an admission that such reference is prior art to the invention. The section headings used should not be construed as necessary limitations. Additionally, any priority documents of this application are incorporated herein by reference in their entirety.

Claims

1. A method for inducing the shedding of cells and molecules from a target epithelium, the method comprising: High-energy focused ultrasound with a mechanical index (MI) greater than 0.4 is used to irradiate the target tissue to generate shear waves within the tissue.

2. The method according to claim 1, wherein, The ultrasonic irradiation includes ultrasonic irradiation of target tissue with ultrasonic energy having a focused beam, the focused beam generating shear waves within the tissue.

3. The method according to claim 1 or 2, wherein, The ultrasonic irradiation includes the application of a focused ultrasonic beam, wherein the ultrasonic beam is arranged as follows: The mechanical index (MI) is generated in the range of 0.4 to 1.8; A beamwidth ranging from 2 mm to 10 mm is generated at the focal point; It is generated with a pulse width ranging from 100 microseconds to 800 microseconds; as well as It is generated at a pulse repetition rate in the range of 4 Hz to 20 Hz.

4. The method according to any one of claims 1-3, wherein, Ultrasonic irradiation includes: The method further includes irradiating the target tissue with ultrasound energy at a first intensity level during a first time period, wherein the first intensity level is higher than the second intensity level.

5. The method according to claim 4, wherein, The first strength level corresponds to a mechanical index (MI) in the range of 0.3 to 1.

8.

6. The method according to any one of claims 4-5, wherein, During the first time period, the target tissue was irradiated with ultrasonic energy, including by ultrasonic irradiation with a focused ultrasonic beam.

7. The method according to any one of claims 4-6, wherein, The second intensity level corresponds to a mechanical index (MI) in the range of 0.1 to 0.

4.

8. The method according to any one of claims 4-7, wherein, During the second time period, the target tissue was irradiated with ultrasound energy, including by ultrasound irradiation with a non-focused ultrasound beam.

9. The method according to any one of claims 4-8, wherein, During the first time period, the target tissue was irradiated with ultrasound energy, including by ultrasound irradiation with repetitive pulses ranging from 2 microseconds to 800 microseconds.

10. The method according to any one of claims 4-9, wherein, During the second time period, the target tissue was irradiated with ultrasound energy, including by repeating ultrasound pulses with a duration ranging from 20 microseconds to 800 microseconds.

11. The method according to any one of claims 9-10, wherein, During the first time period, the target tissue is irradiated with ultrasound energy, including repeated ultrasound pulses at a rate of 1 to 20 times per second.

12. The method according to any one of claims 9-11, wherein, During the second time period, the target tissue is irradiated with ultrasound energy, including repeated ultrasound pulses at a rate of 1 to 100 times per second.

13. The method according to any one of claims 1-11, wherein, The duration of the first time period is in the range of 1 minute to 15 minutes.

14. The method according to any one of claims 1-13, wherein, The duration of the second time period is in the range of 3 to 30 minutes.

15. The method according to any one of claims 4-12, wherein, During the second time period, the target tissue is irradiated with ultrasound at a second intensity level, including the delivery of an ultrasound contrast agent to the target tissue prior to the second time period of ultrasound irradiation with ultrasound at a second intensity level.

16. The method according to any one of claims 4-15, wherein, During the second time period, ultrasound energy is used to irradiate the target tissue, including the delivery of ultrasound contrast agents to the target tissue during the second time period.

17. The method according to any one of claims 4-16, wherein, The duration of the second time period is in the range of 10 to 20 minutes.

18. The method according to any one of claims 4-16, wherein, The target tissue includes the pancreas.

19. The method of claim 18, further comprising: Administer a drug to the patient to induce pancreatic secretion; And to collect cytopathological samples by collecting pancreatic secretions.

20. The method according to any one of claims 4-18, wherein, The target tissue includes those selected from the following body parts: mediastinum; pleura; Pericardium; peritoneum; lung; breast; Salivary glands; Meninges; pancreas; Pancreatic duct; Pancreatic cysts; prostate; kidney; liver; Bladder; and Ovary.

21. The method according to claim 20, wherein, The target tissue includes the prostate, and the method includes collecting one or both of urine and semen.

22. The method of claim 20 or 21, comprising promoting urination in the patient by administering a diuretic substance to the patient and inducing the patient to consume fluids, or by one or both of these methods.

23. A method for inducing exfoliation, the method comprising irradiating a target tissue with ultrasonic energy having a mechanical index (MI) in the range of 1.2 to 1.

8.

24. The method according to claim 23, wherein, Ultrasonic irradiation of target tissues via ultrasonic energy includes ultrasonic irradiation via focused ultrasonic beams.

25. The method according to any one of claims 23-24, wherein, Ultrasonic irradiation of target tissues via ultrasonic energy includes ultrasonic irradiation with repetitive pulses ranging from 20 microseconds to 800 microseconds.

26. The method of claim 25, wherein, Ultrasonic irradiation of target tissue using ultrasonic energy involves repeating ultrasonic pulses at a rate of 1 to 50 times per second.

27. An ultrasonic energy source programmed to perform ultrasonic irradiation using an ultrasonic irradiation procedure, the procedure comprising: Select program parameters to generate shear waves that induce or assist cell shearing; and During the first time period, focused ultrasound energy designed to generate shear waves in the irradiated tissue was used for ultrasound irradiation at a first intensity level.

28. The ultrasonic energy source according to claim 27, wherein, After or before the ultrasound irradiation: During a second time period, ultrasonic irradiation is performed using unfocused ultrasonic energy at a second intensity level, wherein the second intensity level is lower than the first intensity level.

29. A program for controlling an ultrasound source, the program comprising: During the first time period, ultrasonic irradiation was performed using ultrasonic energy of the first intensity level. Subsequently During the second time period, ultrasonic irradiation was performed using ultrasonic energy at the second intensity level. in The first intensity level is higher than the second intensity level.

30. A method for programming an ultrasound source, the method comprising: The ultrasound source is programmed to perform ultrasound irradiation using ultrasound energy of a first intensity level during a first time period. The ultrasound source is programmed to perform ultrasound irradiation using a second intensity level of ultrasound energy during the second time period. in The first intensity level is higher than the second intensity level.

31. A separated sample of bodily fluid obtained from an ultrasound-irradiated prostate of a subject, wherein, The separated sample contains exfoliated cells in sufficient quantity for the analysis and detection of abnormal cells.

32. The body fluid separation sample according to claim 31, comprising one or both of urine and semen.

33. The separated sample of body fluid according to claim 31 or 32, comprising adjacent cells from the prostate.

34. The separated body fluid sample according to claim 33, wherein, The adjacent cells include fragments or cell sheets of epithelial tissue.

35. The separated body fluid sample according to claim 34, comprising more than one epithelial tissue fragment.

36. The separated sample of body fluid according to claim 34 or 35, comprising intact epithelial tissue fragments from an organ.

37. A separated sample of body fluid according to any one of the preceding claims, wherein, The separated sample was obtained by irradiating the prostate with high-intensity focused ultrasound.

38. An ultrasound device programmed to irradiate the prostate, comprising at least one probe for irradiating the prostate, wherein the at least one probe comprises one or more of the following: a transrectal ultrasound probe, a transperineal ultrasound probe, and an ultrasound catheter probe.

39. The ultrasound device programmed to irradiate the prostate with ultrasound according to claim 38, including an electroejaculation device.

40. The ultrasound device programmed to irradiate the prostate with ultrasound according to claim 38 or 39, including a prostate massager.

41. The ultrasound device programmed to irradiate the prostate according to any one of claims 38-39, programmed to: Select program parameters to generate shear waves used to induce or facilitate cell shearing; and During the first time period, focused ultrasound energy designed to generate shear waves in the irradiated tissue was used for ultrasound irradiation at a first intensity level.

42. The ultrasound device programmed to irradiate the prostate with ultrasound according to claim 41, wherein, After or before the ultrasound irradiation: During a second time period, ultrasonic irradiation is performed using unfocused ultrasonic energy at a second intensity level, wherein the second intensity level is lower than the first intensity level.

43. A method for inducing exfoliation, the method comprising irradiating a target tissue with ultrasonic energy in the range of 1.2 to 1.8 by means of an ultrasonic energy.

44. The method according to claim 43, wherein, Ultrasonic irradiation of target tissues via ultrasonic energy includes ultrasonic irradiation via focused ultrasonic beams.

45. The method according to any one of claims 43-44, wherein, Ultrasonic irradiation of target tissues via ultrasonic energy includes ultrasonic irradiation with repetitive pulses ranging from 20 microseconds to 800 microseconds.

46. ​​The method according to claim 45, wherein, Ultrasonic irradiation of target tissue using ultrasonic energy involves repeating ultrasonic pulses at a rate of 1 to 50 times per second.

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