Method for preparing and testing functional water capable of enhancing cell penetrating power and application of functional water

Abstract

The invention provides a method for preparing functional water capable of enhancing cell penetrating capacity. The method comprises: preparing a ceramic by melting natural clay and iron-containing sand; immersing the ceramic in water for a first predetermined time at a predetermined temperature according to a predetermined ceramic-water weight ratio; and carrying out ultrasonic treatment on the ceramic in water for a second preset duration at a preset temperature and a preset ultrasonic power. The ultrasonically treated water is applied to flow through a column filled with a ceramic at a predetermined temperature to obtain functional water. Water flows through the cylinder at a predetermined water flow to ceramic weight ratio or at a third predetermined time required for water per volume to pass through the ceramic per weight. The ceramic causes a structural change in at least some of the water molecules of the water.

Description

[0001] Cross-references to related patent applications

[0002] Pursuant to Section 35 of the United States Code, Section 119(e), this PCT application claims priority to and the benefit of U.S. Provisional Patent Application No. 63 / 322,378, filed on March 22, 2022, entitled “Methods and Applications of Functional Water with Enhanced Cell Penetration Ability” (author Yungfeng Lu), the entire contents of which are incorporated herein by reference. Technical Field

[0003] The present invention relates generally to water, and more particularly to methods for preparing and testing functional water having enhanced cell penetration ability and applications thereof. Background Art

[0004] The background description provided herein is for the purpose of presenting the background of the present invention in general. The subject matter discussed in the background of the present invention should not be considered as prior art simply because it is mentioned in the background of the present invention. Similarly, the problems mentioned in the background of the present invention or associated with the subject matter in the background of the present invention should not be considered as having been previously recognized in the prior art. The subject matter in the background of the present invention merely represents different methods, which themselves may also be inventions.

[0005] Water is a key component of all living organisms. The human body is mostly water. Water makes up about 75% of the body weight of infants, 50% to 60% of the body weight of adult men and women, and as low as 45% of the body weight of the elderly. All chemical reactions and physiological processes, from the absorption and processing of nutrients to the elimination of metabolic waste products, take place in aqueous solutions. The average adult ingests about 2500mL of aqueous fluids per day (mostly from the digestive tract), of which 230mL is produced through metabolism. About the same volume of water leaves the body each day, primarily in the form of urine, as well as through evaporation from the skin and as air expelled from the lungs.

[0006] According to the Centers for Disease Control and Prevention (CDC), drinking water can prevent dehydration. Dehydration can cause mood changes due to unclear thinking, cause the body to overheat, and lead to constipation and kidney stones. In addition to water, there are many beverages available as part of a healthy diet. In particular, functional water is water enhanced with supplements such as vitamins, minerals, fruits, and herbs, which provide additional health benefits and other functional benefits. The past few decades have witnessed rapid growth in the functional water market due to the shift in consumer preferences from sugary and carbonated drinks to water and the desire for more than just hydration. In particular, alkaline water, electrolyzed reduced water, and hydrogen-rich water have gained increasing attention due to their potential anti-inflammatory and antioxidant properties.

[0007] Therefore, there is still a need in the art to address the above-mentioned defects and deficiencies. Summary of the invention

[0008] In one aspect, the present invention relates to a method for preparing aquaporin water with enhanced cell penetration ability. In one embodiment, the method includes: preparing ceramics by melting natural clay and iron-containing sand; immersing ceramics in water for a first predetermined time at a predetermined ceramic-water weight ratio at a predetermined temperature, and ultrasonically treating ceramics in water for a second predetermined time at a predetermined temperature with a predetermined ultrasonic power; and applying ultrasonically treated water to flow through a cylinder filled with ceramics at a predetermined temperature, thereby obtaining aquaporin water, wherein water flows through the cylinder at a predetermined water flow-ceramic weight ratio or at a third predetermined time required for a unit volume of water to pass through a unit weight of ceramics. Ceramics cause structural changes in at least some water molecules of water.

[0009] In one embodiment, the water molecules of aquaporin water have a HOH bond angle α, and the HOH bond angle α1 of the water molecules after the change is greater than the conventional HOH bond angle α0 of the water molecules before the treatment. In one embodiment, the HOH bond angle α1 is about 120°, while the conventional bond angle α0 is 104.45°.

[0010] In one embodiment, the method further includes: verifying the various capabilities of aquaporin water by: preparing a culture medium by placing a growth-promoting substance in the aquaporin water, and filtering the aquaporin water to remove precipitates and bacteria in the aquaporin water; preparing a control culture medium by placing a growth-promoting substance in deionized water; after preparing the culture medium and the control culture medium, placing cells of a living subject in the culture medium and the control culture medium, respectively; and measuring the cells in the culture medium and the control culture medium using corresponding kits to determine the capabilities of the aquaporin water.

[0011] In one embodiment, the abilities of aquaporin water include: increasing cell viability; protecting cells from oxidative damage; maintaining telomere length of cells; and down-regulating genes associated with inflammation and aging in a living subject.

[0012] In one embodiment, the ceramic is a Tadanoumi ceramic, the predetermined ceramic-water weight ratio is a ceramic-water ratio of 0.01-0.1 g / ml, the predetermined temperature is room temperature, the first predetermined time period is 18-36 hours, the predetermined ultrasonic power is 100 watts, the second predetermined time period is 3-7.5 minutes, the predetermined water flow-ceramic weight ratio is 0.01 liters / minute / gram, and the third predetermined time required for 1 liter of water to pass through 1 gram of ceramic is 0.01 seconds.

[0013] In one embodiment, the growth promoting substance comprises: minimum essential medium (MEM) as a base medium; and fetal bovine serum (FBS) and penicillin / streptavidin (P / S) as supplementary substances.

[0014] Another aspect of the present invention relates to aquaporin water prepared by the method disclosed above. In certain embodiments, the water molecules of the aquaporin water have a HOH bond angle α, and the HOH bond angle α1 of the water molecules after the change is greater than the conventional HOH bond angle α0 of the water molecules of the water before the treatment. In one embodiment, the HOH bond angle α1 is about 120°, while the conventional bond angle α0 is 104.45°.

[0015] Aquaporin water prepared by the method disclosed above can be used for various applications. For example, another aspect relates to a method for strengthening the elimination of metabolic waste in a living subject, comprising: preparing aquaporin water as described above; and providing the aquaporin water to a living subject. In certain embodiments, aquaporin water is used to strengthen the elimination of metabolic waste from living subject cells. In one embodiment, the metabolic waste comprises hydrogen peroxide.

[0016] Yet another aspect relates to a method comprising: preparing aquaporin water as described above; and providing the aquaporin water to a living subject. In certain embodiments, the aquaporin water can be used to: improve cell viability of a living subject; maintain telomere length of cells of a living subject; and / or anti-inflammatory to downregulate genes of a living subject.

[0017] In another aspect, the present invention relates to a method for preparing functional water with enhanced cell penetration ability. In one embodiment, the method includes: preparing ceramics by melting natural clay and iron-containing sand; immersing the ceramics in water for a first predetermined time at a predetermined ceramic-water weight ratio at a predetermined temperature to obtain functional water, and ultrasonically treating the ceramics in water for a second predetermined time at a predetermined temperature at a predetermined ultrasonic power; and applying ultrasonically treated water to flow through a column filled with ceramics at a predetermined temperature, thereby obtaining functional water, wherein the water flows through the column at a predetermined water flow-ceramic weight ratio or at a third predetermined time required for a unit volume of water to pass through a unit weight of ceramics. The ceramic causes structural changes in at least some of the water molecules of the water.

[0018] In one embodiment, the method further includes verifying the ability of the functional water by placing cells of a living subject in functional water as a culture medium; placing cells of a living subject in deionized water as a control culture medium; and measuring the cells in the culture medium and the control culture medium using a corresponding kit to determine the ability of the functional water. In one embodiment, the ability of the functional water includes: improving cell viability; protecting cells from oxidative damage; maintaining cell telomere length; and downregulating genes associated with inflammation and aging in a living subject.

[0019] In one embodiment, the method further comprises: placing a growth promoting substance in the culture medium before placing the cells in the functional water; and filtering the culture medium to remove sediment and bacteria in the functional water. In one embodiment, the growth promoting substance comprises: minimum essential medium (MEM) as a basal culture medium; and fetal bovine serum (FBS) and penicillin / streptavidin (P / S) as supplementary substances.

[0020] In one embodiment, the ceramic is Tadanoumi ceramic and the functional water is aquaporin water. In one embodiment, the predetermined ceramic-water weight ratio is a ceramic-water ratio of 0.01-0.1 g / ml. In one embodiment, the predetermined temperature is room temperature, the first predetermined duration is 18-36 hours, the predetermined ultrasonic power is 100 watts, the second predetermined duration is 3-7.5 minutes, the predetermined water flow-ceramic weight ratio is 0.01 liters / minutes / grams, and the third predetermined time required for 1 liter of water to pass through 1 gram of ceramic is 0.01 seconds.

[0021] Another aspect of the present invention relates to aquaporin water prepared by the method disclosed above. In certain embodiments, the water molecules of the aquaporin water have a HOH bond angle α, and the HOH bond angle α1 of the water molecules after the change is greater than the conventional HOH bond angle α0 of the water molecules of the water before the treatment. In one embodiment, the HOH bond angle α1 is about 120°, while the conventional bond angle α0 is 104.45°.

[0022] Another aspect of the present invention relates to an aqueous solution, which includes the water channel protein water disclosed above, wherein the volume of the changed water molecules in the aqueous solution is V1; the volume of the unchanged water molecules in the aqueous solution is V0; the volume of the aqueous solution is V, V=V1+V0, and V1 / V0>1.

[0023] The functional water prepared by the method disclosed above can be used for various applications. For example, another aspect relates to a method for enhancing the elimination of metabolic waste in a living subject, comprising: preparing the functional water as described above; and providing the functional water to the living subject. In certain embodiments, the functional water can be used to enhance the elimination of metabolic waste from the cells of the living subject. In one embodiment, the metabolic waste includes hydrogen peroxide.

[0024] Yet another aspect relates to an aqueous solution comprising: a certain number of first water molecules having a first volume V1, which have a first average HOH bond angle α1 between two HO bonds of each first water molecule; and a certain number of second water molecules having a second volume V0, which have a second average HOH bond angle α0 between two HO bonds of each second water molecule.

[0025] In one embodiment, the first average HOH bond angle α1 is greater than the second average HOH bond angle α0. In one embodiment, the first average HOH bond angle α1 is 120°, and the second average HOH bond angle α0 is about 104.45°.

[0026] In one embodiment, the volume of the aqueous solution is V, wherein V=V1+V0, and V1 / V0>1.

[0027] These and other aspects of the present invention will become apparent from the following description of preferred embodiments in conjunction with the accompanying drawings, but they may be varied and modified without departing from the spirit and scope of the novel concepts of the present invention. BRIEF DESCRIPTION OF THE DRAWINGS

[0028] The accompanying drawings illustrate one or more embodiments of the present invention and, together with the written description, serve to explain the principles of the present invention. Wherever possible, the same reference numerals are used throughout the drawings to refer to the same or similar elements of a particular embodiment.

[0029] Figure 1 A flow chart showing a method for preparing functional water with enhanced cell penetration ability according to certain embodiments of the present invention.

[0030] Figure 2 It is shown that according to some embodiments of the present invention, verification is performed by Figure 1 A flow chart showing the process of preparing functional water with capacity.

[0031] Figure 3 The geometry of an exemplary water molecule according to some embodiments of the present invention is schematically shown.

[0032] Figure 4AThe potential relationship between the energy spectrum of a water molecule and the HOH bond angle based on the proposed model according to some embodiments of the present invention is schematically and qualitatively shown.

[0033] Figure 4B A proposed three-dimensional model indicating the volume and effective area of ​​a water molecule in relation to the HOH bond angles according to certain embodiments of the present invention is shown.

[0034] Figure 5 Characteristics of AQP water with higher aquaporin permeability are shown, where (A) shows the relative AQP permeability (AQP pf) values ​​of AQP water prepared from distilled water (dw) and dw; (B) shows photographs of clay (top) and Tadanoumi ceramic (bottom); and (C) shows the proliferation of human skin cells in control culture medium and ceramic-treated culture medium.

[0035] Figure 6 Shown are (A) X-ray diffraction (XRD) patterns and (B) thermogravimetric analysis (TGA) of Tadanoumi ceramics according to certain embodiments of the present invention.

[0036] Figure 7 Shown are (A) relative cell viability of AML-12 cultured in DI-water and AQP-water, with P value: *<0.05; and (B) relative cell death rate of HepG2 cultured in DI-water and AQP-water.

[0037] Figure 8 Shown are (A) relative T / S ratios of cells cultured in medium prepared with DI water or AQP water as determined by qPCR, wherein P value: *<0.05; and (B) RTL ratios of cells cultured in medium prepared with DI water or AQP water as determined by qPCR.

[0038] Fig. 9 Shown are the fold changes in gene expression determined by RT-PCR for cells cultured in AQP water relative to cells cultured in control water. DETAILED DESCRIPTION

[0039] The present invention will now be described more fully below in conjunction with the accompanying drawings, in which exemplary embodiments of the present invention are shown. However, the present invention can be implemented in many different forms and should not be construed as being limited to the embodiments set forth herein. On the contrary, these embodiments are provided to make this specification comprehensive and complete and to fully convey the scope of the present invention to those skilled in the art. The same reference numerals refer to the same elements throughout.

[0040] The terms used in this specification generally have their ordinary meanings in the art, in the context of the present invention, and in the specific context in which each term is used. Certain terms used to describe the present invention will be discussed below or elsewhere in this specification to provide additional guidance to practitioners regarding the present invention. For convenience, certain terms may be highlighted, for example, using italics and / or quotation marks. The use of highlighted content has no effect on the scope and meaning of the terms. In the same context, the scope and meaning of the terms are the same, whether or not they are in a highlighted form. It should be appreciated that the same thing can be expressed in a variety of ways. Therefore, alternative language and synonyms can be used for any one or more terms discussed herein, and whether the terms are elaborated or discussed in detail in this article does not have any special meaning. Synonyms for certain terms are provided. The enumeration of one or more synonyms does not exclude the use of other synonyms. The examples adopted anywhere in this specification (including examples of any terms discussed herein) are only illustrative and are by no means a limitation on the scope and meaning of the present invention or any exemplary term. Similarly, the present invention is not limited to the various embodiments given in this specification.

[0041] It should be understood that, as used in the description herein and throughout the claims that follow, the meanings of "a," "an," and "the" include plural references unless the context clearly dictates otherwise. In addition, it should be understood that when an element is referred to as being "on another element," it can be directly on the other element, or there can be intervening elements between them. Conversely, when an element is referred to as being "directly on another element," there are no intervening elements. As used herein, the term "and / or" includes any and all combinations of one or more of the listed associated items.

[0042] It should be understood that, although the terms "first", "second", "third", etc. may be used herein to describe various elements, components, regions, layers and / or sections, these elements, components, regions, layers and / or sections should not be limited by these terms. These terms are only used to distinguish a certain element, component, region, layer or section from another element, component, region, layer or section. Thus, the first element, component, region, layer or section discussed below may be referred to as a second element, component, region, layer or section without departing from the teachings of the present invention.

[0043] In addition, relative terms such as "lower" or "bottom" and "upper" or "top" may be used herein to describe the relationship of one element to another element, as shown in the figure. It should be understood that, in addition to the directions shown in the figures, relative terms are also intended to cover different directions of the device. For example, if the device in one of the figures is turned over, the element described as being on the "lower" side of the other elements will be facing the "upper" side of the other elements. Therefore, depending on the specific orientation of the drawings, the exemplary term "lower" may include both "lower" and "upper" directions. Similarly, if the device in one of the drawings is turned over, the element described as being "below" or "directly below" other elements should be facing "above" these other elements. Therefore, the exemplary terms "below" or "below" may include both upper and lower directions.

[0044] It should also be understood that the terms "include" and / or "it includes", "includes" and / or "it includes", "have" and / or "it has", "carry" and / or "it carries", "contain" and / or "it contains" or "involve" and / or "it involves" etc. are open-ended, i.e., refer to including but not limited to. When used in this specification, these terms indicate the presence of the described features, regions, wholes, steps, operations, elements and / or parts, but do not exclude the presence or addition of one or more other features, regions, wholes, steps, operations, elements, parts and / or their combinations.

[0045] Unless otherwise defined, all terms (including technical and scientific terms) used herein have the same meaning as those commonly understood by those skilled in the art to which the present invention belongs. It should also be understood that terms (such as those defined in commonly used dictionaries) should be understood to have the same meaning as they have in the context of the relevant art and this specification, and should not be understood in an idealized or overly formal sense unless clearly defined herein.

[0046] As used in this specification, "about", "approximately", "approximately" or "substantially" generally means within 20% (preferably within 10%, more preferably within 5%) of a given value or range. The numerical values ​​given herein are approximate, meaning that if not explicitly stated, the term "about", "approximately", "substantially" can be inferred.

[0047] As used in this specification, the phrase at least one of A, B, and C should be construed to mean a logical (A or B or C), using a non-exclusive logical OR. As used herein, the term "and / or" includes any and all combinations of one or more of the associated listed items.

[0048] The following description is merely exemplary in nature and is not intended to limit the invention and its application or use. The broad teachings of the invention may be implemented in various forms. Therefore, although the invention includes specific examples, the true scope of the invention should not be limited, because other modifications will become apparent when studying the drawings, the specification and the following claims. For clarity, the same reference numerals will be used in the drawings to identify similar elements. It should be understood that one or more steps in the method may be performed in a different order (or simultaneously) without changing the principles of the invention.

[0049] As mentioned above, the past few decades have witnessed a rapid growth in the functional water market. One type of functional water prepared may be aquaporin (AQP) water.

[0050] Aquaporins (AQP): AQP water is a new type of functional water that enables faster water transport across cell membranes. Cells are the basic structural and functional units of life, including cytoplasm surrounded by cell membranes. The cell membrane provides a protective barrier around the cell while regulating which substances or molecules can or cannot enter.

[0051] Ion channels that regulate the transport of ions across cell membranes (e.g., calcium ions (Ca 2+ ) channels, potassium ions (K + ) channels, sodium ions (Na + ) channels and proton (H + ) channels), the transport of water molecules across cell membranes (alone or in groups) is mediated by water channels (also called aquaporins) and is driven by the osmotic pressure across the cell membrane. Aquaporins are intrinsic membrane proteins and are present in all living organisms. Aquaporins form pores in the cell membrane to facilitate water transport between cells. The existence of aquaporins was hypothesized 60 years ago, and it was not until 1992 that the first aquaporin (aquaporin 1) was discovered by Peter Agre of Johns Hopkins University. Peter Agre received the 2003 Nobel Prize in Chemistry for his pioneering work in the discovery of aquaporins.

[0052] Currently, in mammals, 13 aquaporins (AQP0-AQP12) have been identified, and they are traditionally divided into three subgroups: (1) conventional AQPs (AQP0, 1, 2, 5, 6, and 8) that are primarily used for water transport; (2) aquaglycerol channel proteins (AQP3, 7, 9, and 10) (which transport neutral solutes such as glycerol, urea, ammonia, and carbon dioxide in addition to water); and (3) superaquaporins (AQP11 and 12) (the details of which remain to be elucidated). In addition, hydrogen peroxide (H2O2) and water (H2O) have similar dipole moments and hydrogen bonding abilities. Like water, hydrogen peroxide is also transported across cell membranes by aquaporins (a new subset of aquaporins including AQP3, 8, 9, and 11, called peroxiporins). It is important to note that hydrogen peroxide is toxic; effective removal of hydrogen peroxide from the body is critical to reducing oxidative stress, anti-aging, and preventing and treating many diseases.

[0053] AQP AQP water is a new type of functional water prepared according to certain embodiments of the present invention. Compared with ordinary water or other functional water, AQP water has a unique function that allows water to pass through aquaporins and cross cell membranes more efficiently, thereby achieving faster hydration and effectively removing toxic compounds (such as hydrogen peroxide) from the human body.

[0054] Certain aspects of the present invention relate to methods for preparing functional water having enhanced cell penetration ability. Figure 1 A flow chart of a method for preparing functional water with enhanced cell penetration ability according to certain embodiments of the present invention is shown. It should be particularly noted that, unless otherwise specified in the present disclosure, the steps of the method can be arranged in a different order and are therefore not limited to Figure 1 in the order shown.

[0055] like Figure 1 As shown, in the method for preparing functional water, in step 110, ceramics are prepared by melting natural clay and iron-containing sand. In some embodiments, the ceramics may also be Tadanoumi It will be discussed further later. Then, in step 120, the ceramic is immersed in water for a first predetermined time at a predetermined ceramic-water weight ratio at a predetermined temperature, and the ceramic is ultrasonically treated in water for a second predetermined time at a predetermined temperature with a predetermined ultrasonic power. In this step, the ceramic causes at least one or some water molecules of the water to change their structure, and reduces the cluster size of the water. In certain embodiments, when the functional water is AQP water, the predetermined ceramic-water weight ratio can be within the ceramic-water ratio range of 0.01-0.1 g / ml. In one embodiment, the predetermined ceramic-water weight ratio is within the ceramic-water ratio range of 0.05-0.1 g / ml. In one embodiment, the predetermined temperature is room temperature, the first predetermined time is 18-36 hours, the predetermined ultrasonic power is 100 watts, and the second predetermined time is 3.0-7.5 minutes. For example, the first predetermined time can be 24 hours, and the second predetermined time can be 5 minutes.

[0056] Once the water is ultrasonically treated, the ultrasonically treated water is applied in step 130 to flow through a column filled with a ceramic of a predetermined temperature (the same type of ceramic as that immersed in the water) to obtain functional water. Specifically, the ultrasonically treated water flows through the column at a predetermined water flow-ceramic weight ratio or at a third predetermined time required for a unit volume of ultrasonically treated water to pass through a unit weight of ceramic. In one embodiment, the predetermined water flow-ceramic weight ratio is 0.01 liters / minute / gram. In one embodiment, the third predetermined time required for 1 liter of water to pass through 1 gram of ceramic is 0.01 seconds.

[0057] Once the functional water is obtained, the capabilities of the functional water can be verified in step 140. In one embodiment, the capabilities of the functional water that can be verified include: improving cell viability; protecting cells from oxidative damage; maintaining cell telomere length; and downregulating genes associated with inflammation and aging in living subjects.

[0058] FIG.2 Figure 2 It is shown that according to some embodiments of the present invention, verification is performed by Figure 1 Flow chart of the process of preparing functional water of the method shown. It should be particularly noted that unless otherwise specified in the present disclosure, the steps of the process can be arranged in a different order and are therefore not limited to Figure 2 in the order shown.

[0059] like Figure 2As shown, in step 210, a culture medium is prepared by placing a growth-promoting substance in functional water, and filtering the functional water to remove sediment and bacteria in the functional water. In one embodiment, the growth-promoting substance includes: minimum essential medium (MEM) as a basal culture medium; and fetal bovine serum (FBS) and penicillin / streptococcus (P / S) as supplementary substances. In step 220, a control culture medium is prepared by placing the growth-promoting substance in deionized water. After preparing the culture medium and the control culture medium, in step 230, cells of a living subject are placed in the culture medium and the control culture medium, respectively, to perform a verification step. In step 240, the cells in the culture medium and the control culture medium are measured using a corresponding kit to determine the capacity of the functional water.

[0060] AQP water was first described by Tadao Kozumi (Cosmic Com Ltd., Hiroshima, Japan) and Yoshichika Kitagawa (Akita Prefectural University, Japan) in "water structure changes induced by ceramics can be detected by increased permeability through aquaporin ”(1) The study was reported in Biochemistry and Biophysics Reports (5, 2016, 353-358), and the entire contents of which are incorporated herein by reference. (See FIG4(B)) AQP water was prepared by immersing the ceramic in distilled or tap water for 24 hours (4 g of ceramic in 50 mL of water, which corresponds to a ceramic-water ratio of 0.08 g / mL). Figure 1 In the method shown, in addition to the immersion step, the AQP water of the present invention is obtained by further ultrasonically treating the ceramic in water after the immersion time, and then further applying the ultrasonically treated water to flow through a column filled with ceramic to filter the water. In this case, the water molecular structure change caused by the Tadanoumi ceramic and the reduced water cluster size can be made more stable, thereby enhancing the stability of the obtained AQP water.

[0061] FIG.3 Figure 3 The geometric shapes of exemplary water molecules according to certain embodiments of the present invention are schematically shown. As described above, by implementing the method for preparing functional water, the water molecules are caused to undergo structural changes. Specifically, Figure 3As shown, initially, the exemplary water molecule 300 in the conventional state 300a has a HOH bond angle α0 (which is the angle defined between two HO bonds 310) and a bond length L0 (which is the length between the hydrogen and oxygen along each HO bond 310), and for conventional water molecules, the conventional bond angle α0 is 104.45° on average and the bond length L0 is However, for the functional water obtained by the above process according to some embodiments of the present invention, at least one or some water molecules in state 300a are induced or changed to state 300b, wherein the HOH bond angle of the functional water molecules in state 300b is changed from α0 to α, wherein α≠α0, and the bond length L of the functional water molecules in state 300b can remain substantially unchanged or change accordingly. In other words, in the process of preparing functional water as described above, the structure of one or more water molecules 300 has been changed or modified.

[0062] In certain embodiments, once water molecules are treated by the above method for preparing functional water, it is observed that the HOH bond angle α of aquaporin water molecules is about α=α1=120.00° on average. Based on this observation, a possible and credible model for the change of HOH bond angle is proposed: (2) Specifically, Figure 4A The potential relationship between the energy spectrum of a water molecule and the HOH bond angle based on the proposed model according to some embodiments of the present invention is schematically and qualitatively shown. Figure 4A As shown, the recommended energy spectrum E of the water molecule related to the HOH bond angle is schematically and qualitatively shown, wherein point A is the first minimum of the energy spectrum curve 400. For conventional water, the water molecule stays in the energy ground state corresponding to point A on the energy spectrum curve 400, which can be characterized by energy E0, and the corresponding conventional bond angle α0 is 104.45°. When preparing functional water according to certain embodiments of the present invention, the energy of the water molecule can be changed to cross the peak point B (which is a relative maximum point) and then reach the next relative minimum point C at the energy spectrum curve 400, where the corresponding HOH bond angle α1 is approximately 120°, so that the changed or modified water molecule can be retained and remain stable for a period of time (e.g., more than one week). It should be noted that the energy spectrum curve 400 shown may include multiple relative energy minima, so that there may be additional relative minimum points D and F, wherein the HOH bond angle α at each point is further greater than the conventional bond angle α0 of 104.45°, respectively.

[0063] In other words, in certain embodiments, as the water molecules are altered or modified, the average HOH bond angle α of the altered or modified water molecules may be greater than the conventional HOH bond angle α0 of 104.45° (eg, points C, D, and F)).

[0064] It has been observed that due to the altered or modified water molecules, functional water can be easily consumed by the human body and provide significant health benefits to humans, which can be further illustrated by the above credible model. (2) Specifically, Figure 4B 3D model proposed according to some embodiments of the present invention is shown, which indicates the volume and effective area of ​​water molecules related to the HOH bond angle. Figure 4B As shown in the middle part, a water molecule is formed by an oxygen atom and two hydrogen atoms, and the two H-0 bonds are in the same plane (hereinafter referred to as the XZ plane on the three-dimensional coordinates), wherein when the HOH bond region rotates in the XZ plane and the YZ plane at the same time, the water molecule is formed in three-dimensional space. Since the conventional HOH bond angle α0 of the conventional water molecule is 104.45° and the HOH bond angle α1 of the changed or modified water molecule is increased to about 120°, the effective area (such as Figure 4B The effective area A0 of a conventional water molecule is approximately 2, and the effective area A1 of the modified water molecule is approximately 2), and based on the zero-order approximation, the effective volume of the water molecules and the corresponding surface S will also decrease accordingly. This results in a decrease in the surface tension of the changed or modified water molecules, as shown by F=a·S. Therefore, harmful free radicals produced by the human body can more easily attach to the modified water molecules and / or be carried by the modified water molecules and eliminated from the body cells. On the other hand, the changed or modified water molecules of functional water can provide enhanced penetration ability because their "effective volume" is smaller. Therefore, the functional water prepared according to certain embodiments of the present invention contains a first number of changed or modified water molecules having a first volume V1 and a first HOH bond angle α1 and a second number of conventional water molecules having a second volume V0 and a second HOH bond angle α0, where α1>α0. In order to be effective, the ratio of V1 / V0 needs to be greater than 1. In other words, V1 / V0>1.

[0065] The unique ability of AQP water to more efficiently pass through human aquaporins was demonstrated in oocytes expressing human aquaporins AQP1, 2, 3, or 5. In short, oocytes in a culture medium were transferred to water. Driven by osmotic pressure, water passed through the aquaporins, causing the cells to swell. The swelling rate of the cells was then measured, and the swelling rate was used to calculate the osmotic water permeability (Pf). It was found that the Pf of tap water treated with Tadanoumi ceramics increased by 26%, 48%, and 21% for oocytes expressing AQP2, AQP3, and AQP5, respectively, compared to tap water (statistically significant p<0.05). Similarly, the Pf of distilled water treated with Tadanoumi ceramics increased by 2.2, 1.2, 1.3, and 1.6 times for oocytes expressing AQP1, AQP2, AQP3, and AQP5, respectively, compared to distilled water, as shown in Figure 2. Figure 5 (A) shown.

[0066] At the same time, it was found that APQ water can also promote the proliferation of normal human skin cells. To verify this, the cell culture medium was first mixed with Tadanoumi (1 g of Cera in 50 ml of medium, equivalent to a Cera-water ratio of 0.02 g / ml) and incubated at room temperature for 24 h. Fully grown cells were trypsinized and incubated at 1.0 × 10 5 These cells were used at a concentration of 10 cells / mL. Cells cultured in control medium or ceramic-treated medium for 48 hours were then treated with Cell Counting Kit-8. The number of viable cells was determined by measuring the absorbance at 450 nm of water-soluble formazan, which is produced by intracellular dehydrogenases. Thus, the proliferation of cells cultured in ceramic-treated medium was 12% higher than that of cells in control medium (statistically significant p<0.05, see Figure 5 (C) ), indicating that the use of ceramic-treated medium promoted cell proliferation.

[0067] Due to the ability of AQP water to more efficiently pass human aquaporins and promote the proliferation of normal human skin cells, AQP water can be used as a consumable aqueous solution. For example, an aqueous solution containing at least 75% by weight of AQP water as disclosed above can be used as a consumable solution for humans.

[0068] Tadanoumi Tadanoumi It is a unique product made by melting natural clay and iron-containing sand from Hiroshima, Japan. Figure 6 (A) X-ray diffraction (XRD) patterns and (B) thermogravimetric analysis (TGA) of Tadanoumi ceramics according to certain embodiments of the present invention are shown. Figure 6(A) X-ray diffraction (XRD, Miniflex II, Rigaku, Cu Ka radiation The crystal structure was examined at 40 kV, 30 mA, and a scan rate of 5° / min. The Tadanoumi ceramics showed a diffraction pattern that did not match any known pattern in the Joint Committee on Powder Diffraction Standards (JCPDS) database. The Tadanoumi ceramics were dense (non-porous) and had a pore size of 0.8 cm by nitrogen adsorption-desorption measurements at 77 K. 2 / g has a negligible surface area. Figure 6 As shown in (B), thermal stability was measured by thermogravimetric analysis (TGA, Netzsch STA 449F3 Jupiter, air flow rate 50 liters / minute, heating rate 10°C / minute). Tadanoumi ceramics showed negligible mass loss at temperatures up to 1000°C, indicating that they have high thermal stability. In addition, it was confirmed by X-ray fluorescence spectroscopy that Tadanoumi ceramics do not contain any detectable heavy metals.

[0069] The functional water prepared by the method disclosed above can be used for various applications. For example, another aspect relates to a method for enhancing the elimination of metabolic waste in a living subject, comprising: preparing the functional water as described above; and providing the functional water to the living subject. In certain embodiments, the functional water can be used to: enhance the elimination of metabolic waste from the cells of the living subject; increase the vitality of the cells of the living subject; maintain the telomere length of the cells of the living subject; and / or anti-inflammatory, to downregulate the genes of the living subject. In one embodiment, the metabolic waste comprises hydrogen peroxide.

[0070] Ability to Improve Cell Viability and Protect Cells from Oxidative Damage: The ability of AQP water to improve cell viability was tested in AML-12 cells, a murine hepatocyte cell line derived from the liver of transgenic mice. Figure 7 Shown are (A) relative cell viability of AML12 cultured in DI-water and AQP-water, with P value: *<0.05; and (B) relative cell death rate of HepG2 cultured in DI-water and AQP-water. Figure 7As shown in (A), control medium was prepared by dissolving Eagle's MEM (EMEM) powder (Sigma Aldrich) in deionized water (referred to as DI-water), and 10% FBS (Gibco) and 1% P / S (Genesee) were supplemented to the control medium. Experimental medium was prepared by dissolving EMEM powder (Sigma Aldrich) in AQP water (expressed as AQP-water), and 10% FBS (Gibco) and 1% P / S (Genesee) were supplemented to the experimental medium. The medium was filtered by a 0.22 μm PES vacuum filtration system (Genesee) to remove any precipitate and bacteria. According to 10,000 cells / well, AML-12 cells were seeded in 96-well plates. Cells were cultured for 24 hours in DI-water medium and AQP-water medium, respectively, and cell viability was measured using a cell titer blue kit (Sigma).

[0071] use

[0072] The following formula calculates the relative viability of cells cultured in AQP-water:

[0073]

[0074] like Figure 7 As shown in (A), cells cultured in AQP-water exhibited 115% cell viability relative to cells cultured in DI-water (statistically significant p<0.05), indicating that AQP-water enhanced cell viability.

[0075] The ability of AQP-water to protect cells from oxidative damage was examined using HepG2, a continuously proliferating cell line derived from human hepatocellular carcinoma. Figure 6(B) Shown, control medium is prepared by dissolving EMEM powder (Sigma Aldrich) in deionized water (expressed as DI-water), and 10% FBS (Gibco) and 1% P / S (Genesee) are supplemented to the control medium. Experimental medium is prepared by dissolving EMEM powder (Sigma Aldrich) in AQP active water (expressed as AQP-water), and 10% FBS (Gibco) and 1% P / S (Genesee) are supplemented to the experimental medium. The culture medium is filtered by 0.22 μm PES vacuum filtration system (Genesee), to remove any precipitate and bacteria. According to 10000 cells / well, HepG2 cells are seeded in 96-well plates. First, cells are cultured in DI-water and AQP-water for 24 hours respectively. Then the cells are cultured for another 24 hours, and H2O2 is added to the culture medium at a final concentration of 250 μM. Cell viability is measured using a cell titer blue kit (Sigma). The relative cell viability of cells cultured in AQP-water with H2O2 treatment was calculated using the following formula:

[0076]

[0077] The viability of cells cultured in AQP-water (74%) was slightly higher than that of cells cultured in DI-water (71%).

[0078] Ability to maintain telomere length: Telomeres are the terminal segments of linear chromosomes, which are nuclear protein complexes composed of tandem arrays of highly conserved G-rich repeat sequences. Telomeres maintain genomic stability by preventing chromosome ends from being recognized as broken DNA and triggering inappropriate responses associated with DNA damage. With cell division and / or oxidative stress, telomere length is eroded until they reach a critical shortening length, at which point the permanent cell cycle is arrested and the cell stops dividing. As an indicator of aging and oxidative stress, telomere length is considered a biomarker of aging and a high-risk factor for age-related diseases.

[0079] The ability of AQP water in maintaining telomere length was detected with HEK-293, a human epithelial cell line derived from fetal kidney. Control culture medium expressed as control water was prepared by dissolving DMEM powder (Sigma Aldrich) in deionized water supplemented with 10% FBS (Gibco) and 1% P / S (Genesee). AQP culture medium expressed as AQP water was prepared by dissolving DMEM powder (Sigma Aldrich) in AQP water supplemented with 10% FBS (Gibco) and 1% P / S (Genesee). Culture medium was filtered through a 0.22 μm PES vacuum filtration system (Genesee) to remove any precipitate and bacteria.

[0080] HEK-293 cells were cultured in control water and AQP water in T25 culture flasks and subcultured every three days. 5×10 DNA fragments from subcultures of 5, 10, 15, and 20 generations (respectively indicated as P5, P10, P15, and P20) were extracted using a DNA extraction kit (Qiagen). 5 The genomic DNA of each cell was collected. Telomere length was measured by quantitative polymerase chain reaction (qPCR) using the telomere primer pair telg (5'-ACACTAAGGTTTGGGTTTGGGTTTGGGTTTGGGTTAGTGT-3') and telc (5'-TGTTAGGTATCCCTATCCCTATCCCTATCCCTATCCCTAACA-3'). The number of genomic genes was measured by qPCR using the β-globin primer pair Fwd 5'-GTGCGAGAGCGTCAGTATTAAG-3' and Rev 5'-TCCCTGTGTGCTCTGCTGCCATACTA-3'.

[0081] Figure 8(A) Relative T / S ratios of cells cultured in medium prepared with DI water or AQP water as determined by qPCR, where P value: *<0.05; and (B) RTL ratios of cells cultured in medium prepared with DI water or AQP water as determined by qPCR. Specifically, the qPCR test was performed using the following thermal cycle program, which consisted of stage 1 (15 minutes at 95°C), stage 2 (15 seconds at 94°C and 15 seconds at 49°C, 2 cycles in total), and stage 3 (15 seconds at 94°C, 10 seconds at 62°C, and 15 seconds at 74°C, 32 cycles in total, with simultaneous signal acquisition). Standard curves of Cq values ​​relative to DNA concentration were constructed for telomeres and genomic DNA, respectively, using MyiQ software (Bio-Rad iQ5 2.0 Standard Edition Optical System Software). The concentrations of telomeres (T) and genomic DNA (S) in the samples were then calculated based on their Cq values. The T / S ratio was used to quantify changes in telomeres during cell growth. RTL (relative telomere length) was also calculated by normalizing the telomere concentration of cells cultured in AQP water relative to the telomere concentration of cells cultured in control water.

[0082] Figure 8 (A) shows the T / S ratio of cells cultured in control water or AQP water after 0, 5, 10, 15 and 20 passages. The T / S ratio of cells continued to decrease with increasing passage number, indicating that telomere length shortened during culture. Cells cultured in AQP water showed a significantly lower rate of decline in the T / S ratio compared to cells cultured in control water. Figure 8 (B) shows the relative telomere length (RTL) of cells cultured with AQP water relative to cells cultured with control water. Figure 6 (B) The findings are consistent, with RTL values ​​increasing with increasing passage number. P5 exhibited an RTL close to 1.0, while PIO, P15, and P20 exhibited significantly higher RTLs (1.56, 1.76, and 2.05, respectively), indicating that AQP water can effectively slow down the shortening process of telomeres. Compared with control water, AQP water increased the telomere length of P10, P15, and P20 cells by approximately 56%, 76%, and 105%, respectively. These results confirm the ability of AQP water to slow down the shortening process of telomeres.

[0083] Ability to downregulate inflammation- and aging-related genes: Nuclear factor-KB (NF-kB) plays a key role in inflammatory diseases. Activation of NF-KB induces the expression of various pro-inflammatory genes, thereby regulating the production of pro-inflammatory cytokines, recruitment of leukocytes, and formation of inflammasomes. Dysregulated NF-kB activation is a characteristic of chronic inflammatory diseases such as diabetes, cancer, and neurodegenerative diseases. Oxidative stress is another key risk factor for many chronic diseases. The Keapl-Nrf2 [Kelch-like ECH-related protein 1-nuclear factor (erythroid-derived 2)-like 2] regulatory pathway is a major inducible defense against oxidative stress, in which Nrf2 mediates the antioxidant response and its inhibitor Keap1 serves as a sensor of oxidative stress.

[0084] The ability of AQP water in down-regulating genes associated with inflammation and aging was detected in the human epithelial cell line HEK-293 from fetal kidney. The control medium, which was expressed as control water, was prepared by dissolving DMEM powder (Sigma Aldrich) in deionized water supplemented with 10% FBS (Gibco) and 1% P / S (Genesee). The AQP medium, which was expressed as AQP water, was prepared by dissolving DMEM powder (Sigma Aldrich) in AQP active water supplemented with 10% FBS (Gibco) and 1% P / S (Genesee). The culture medium was filtered through a 0.22 μm PES vacuum filtration system (Genesee) to remove any precipitate and bacteria.

[0085] HEK-293 cells were cultured in control water and AQP water in T25 culture flasks and subcultured every three days for 27 generations. RNA was extracted from 5x10 5Cellular RNA was extracted from each cell. The isolated cellular RNA was converted into cDNA using a cDNA reverse transcription kit (Qiagen) by reverse transcription polymerase chain reaction (RT-PCR) method using random primers. Representative genes related to inflammation and aging were monitored by polymerase chain reaction (RT-PCR) method using primer pairs NF-kB (Fwd 5'-GCACCCTGACCTTGCCTATTT-3' and Rev 5'-GTCCCAGGCGCCTTGTGAAGC-3'), Nrf2 (Fwd5'-GAGACAGGTGAATTTCTCCCAAT-3' and Rev5'-TTTGGGAATGTGGGCAAC-3') and Keapl (Fwd5'-GGAGGACCACACCAAGCAAGC-3' and Rev5'-GGATGAAGCCAGCACCACCTTG-3'). The GAPDH gene was measured as an internal control using primers Fwd 5′-GCACCGTCAAGGCTGGAAAC-3′ and Rev 5′-TGGTGAAGACGCCAGTGGA-3′.

[0086] The PCR test was performed using a thermal cycle program consisting of stage 1 (95°C for 15 minutes), stage 2 (94°C for 15 seconds and 49°C for 15 seconds, 2 cycles), and stage 3 (94°C for 15 seconds, 62°C for 10 seconds, and 74°C for 15 seconds, 32 cycles, with simultaneous signal acquisition). The gene expression levels of GAPDH (glyceraldehyde-3-phosphate dehydrogenase), NF-kB, Nrf2, and Keap1 were calculated using MyiQ software (Bio-Rad iQ52.0 Standard Edition Optical System Software). The DNA amounts of cells cultured in AQP water and control water were calculated based on their Cq values ​​and normalized with GAPDH expression.

[0087] Fig. 9 The fold change in gene expression of cells cultured in AQP water relative to cells cultured in control water as determined by RT-PCR is shown. Specifically, Fig. 9 The fold changes in gene expression for cells cultured in AQP water for 27 generations compared to cells cultured in control water are shown. AQP water inhibited NF-kB gene expression by 5-fold compared to cells cultured in control water, indicating an anti-inflammatory effect. In addition, cells cultured in AQP water showed a 0.5-fold decrease in Nrf2 expression. In addition, AQP water enhanced the expression of the Nrf2 inhibitor Keapl by 0.7-fold compared to control water. This result suggests that AQP water can help release oxidative stress during cell aging.

[0088] Potential functions and benefits of AQP water: The above studies have shown that AQP water passes through aquaporins faster than ordinary water and exhibits antioxidant and anti-inflammatory capabilities as well as the ability to preserve telomeres. The mechanism of action of AQP water remains unclear and further research is needed. Kozumi and Kitagawa proposed that the enhanced permeability of AQP water may be attributed to Tadanoumi ceramics, which induce structural changes in at least one or some water molecules and reduce the size of water clusters. Based on the role of aquaporins, which can transport water and metabolites across cell membranes, the observed functions of AQP water may be attributed to faster penetration of AQP water, which can more effectively remove metabolic waste products (such as hydrogen peroxide).

[0089] AQP water prepared by the methods disclosed above can be used in various applications. For example, another aspect relates to a method for enhancing the elimination of metabolic waste products in a living subject, comprising: preparing AQP water as described above; and providing the AQP water to a living subject. In certain embodiments, the AQP water is used to enhance the elimination of metabolic waste products from cells of a living subject. In one embodiment, the metabolic waste products include hydrogen peroxide.

[0090] Hydrogen peroxide is the main component of reactive oxygen species (ROS), which are mainly generated as a byproduct of cellular metabolism through the electron transport chain in mitochondria and cytochrome P450. Another major source is enzymatic reactions mediated by oxidases (e.g., NAPDH oxidase), which are ubiquitous in various cells (especially phagocytes and endothelial cells). In humans and other organisms, hydrogen peroxide is mainly removed and decomposed into water and oxygen by catalase. Catalase is the most abundant antioxidant enzyme and is widely present in the liver, erythrocytes, and alveolar epithelial cells.

[0091] Due to its cytotoxicity, hydrogen peroxide is not only used as a weapon against pathogens (e.g., bacteria), it is also used as a signaling molecule for many physiological processes. If the generation and elimination of hydrogen peroxide are unbalanced, local or systemic accumulation of hydrogen peroxide will result, thereby causing oxidative stress and apoptosis through DNA damage, lipid peroxidation and protein oxidation. Expansion studies have linked ROS to diseases such as cancer, insulin resistance, diabetes, cardiovascular disease, atherosclerosis, aging, gout and chronic inflammation. Achieving effective transport of hydrogen peroxide across cell membranes and its subsequent decomposition into water and oxygen is essential for reducing oxidative stress and maintaining healthy homeostasis. In this regard, the inventors believe that AQP water is a truly unique functional water that can bring significant benefits to human health.

[0092] The above-mentioned functional water can be used in various applications. For example, in another aspect, an aqueous solution can be prepared using functional water. In certain embodiments, for example, the aqueous solution may include: a certain number of first water molecules having a first volume V1 (i.e., functional water having changed or modified water molecules), which have a first average HOH bond angle α1 (i.e., about 120°) between two HO bonds of each first water molecule; and a certain number of second water molecules having a second volume V0 (i.e., conventional water molecules), which have a second average HOH bond angle α0 (e.g., 104.45°) between two HO bonds of each second water molecule. In one embodiment, the first average HOH bond angle α1 (e.g., 120°) is greater than the second average HOH bond angle α0 (i.e., 104.45°). In one embodiment, the volume of the aqueous solution is V, where V=V1+V0, and V1 / V0>1.

[0093] The above description of exemplary embodiments of the invention has been presented for purposes of illustration and description only and is not intended to be exhaustive or to limit the invention to the precise form disclosed.Many modifications and variations are possible in light of the above teachings.

[0094] These embodiments are selected and described in order to illustrate the principles of the present invention and its practical application, so that other persons skilled in the art can utilize the present invention and various embodiments, and these embodiments have various modifications suitable for the specific use expected. For those skilled in the art to which the present invention belongs, alternative embodiments will become apparent without departing from the spirit and scope of the present invention. Therefore, the scope of the present invention is limited by the appended claims, rather than by the above description and the exemplary embodiments described therein.

[0095] Several references, including patents, patent applications, and various publications, are cited and discussed in the description of the present invention. These references are cited and / or discussed merely to clarify the description of the present invention, and no admission is made that any of these references are "prior art" to the invention described herein. The entire contents of all references cited and discussed in this specification are incorporated herein by reference, and to the same extent as if each reference were individually incorporated by reference.

[0096] References

[0097] 1.Kozumi T,Kitagawa Y.,"Water structure changes induced by ceramicscan be detected by increased permeability through aquaporin,Biochem.andBiophys.Rep.(2016Jan.8,5:353-358),doi:10.1016 / j.bbrep.2016.01.002.

[0098] 2.Private communications with Dr.Tim Tingkang Xia,May 30-June 1,2022.

Claims

1. A method for preparing aquaporin water having enhanced cell penetration ability, comprising: Ceramics are prepared by melting natural clay and iron-containing sand; immersing the ceramic in water at a predetermined temperature and a predetermined ceramic-water weight ratio for a first predetermined period of time, and sonicating the ceramic in the water at the predetermined temperature and a predetermined ultrasonic power for a second predetermined period of time; and applying the ultrasonically treated water to flow through a column filled with the ceramic at the predetermined temperature, thereby obtaining the aquaporin water, wherein the water flows through the column at a predetermined water flow-ceramic weight ratio or at a third predetermined time required for a unit volume of water to pass through a unit weight of ceramic; The ceramic causes structural changes in at least some of the water molecules of the water. 2 . The method according to claim 1 , wherein the water molecules of the aquaporin water have a HOH bond angle α, and the HOH bond angle α1 of the altered water molecules is greater than the conventional HOH bond angle α0 of the water molecules of the water before treatment.

3. The method of claim 2, wherein the HOH bond angle α1 is about 120° and the conventional bond angle α0 is 104.45°.

4. The method according to claim 1, further comprising: The various capabilities of the aquaporin water were verified by the following methods: preparing a culture medium by placing a growth promoting substance in the aquaporin water, and filtering the aquaporin water to remove precipitates and bacteria in the AQP water; A control medium is prepared by placing the growth promoting substance in deionized water; After preparing the culture medium and the control culture medium, placing cells of a living subject in the culture medium and the control culture medium, respectively; And using a corresponding kit to measure the cells in the culture medium and the control culture medium to determine the water capacity of the aquaporin.

5. The method according to claim 4, wherein the capacity of the aquaporin water comprises: increasing the viability of the cells; Protect the cells from oxidative damage; maintain the telomere length of the cells; and downregulating genes associated with inflammation and aging in the living subject.

6. The method of claim 1 , wherein the ceramic is a Tadanoumi ceramic, the predetermined ceramic-water weight ratio is a ceramic-water ratio of 0.01-0.1 g / ml, the predetermined temperature is room temperature, the first predetermined time period is 18-36 hours, the predetermined ultrasonic power is 100 watts, the second predetermined time period is 3-7.5 minutes, the predetermined water flow-ceramic weight ratio is 0.01 liters / minute / gram, and the third predetermined time required for 1 liter of water to pass through 1 gram of ceramic is 0.01 seconds.

7. The method of claim 1, wherein the growth promoting substance comprises: Minimum Essential Medium (MEM) as basal medium; As well as fetal bovine serum (FBS) and penicillin / streptavidin (P / S) as supplements.

8. Aquaporin water prepared by the method according to claim 1, wherein water molecules of the aquaporin water have a HOH bond angle α, and the HOH bond angle α1 of the altered water molecules is greater than the conventional HOH bond angle α0 of the water molecules of the water before treatment. 9 . The aquaporin water according to claim 8 , wherein the HOH bond angle α1 is about 120°, and the conventional bond angle α0 is 104.45°.

10. An aqueous solution having the aquaporin water according to claim 8, wherein: The volume of the changed water molecules in the aqueous solution is V1; the volume of the unchanged water molecules in the aqueous solution is V0; the volume of the aqueous solution is V, V=V1+V0, and V1 / V0>1.

11. A method for enhancing the elimination of metabolic waste in a living subject, comprising: preparing the aquaporin water according to claim 1; and providing the aquaporin water to the living subject, wherein the aquaporin water is used to enhance the elimination of the metabolic waste from the cells of the living subject.

12. The method of claim 11, wherein the metabolic waste product comprises hydrogen peroxide.

13. A method for increasing the viability of cells in a living subject, comprising: preparing the aquaporin water according to claim 1; and providing the aquaporin water to the living subject, wherein the aquaporin water is used to improve the cell viability of the living subject.

14. A method of protecting cells of a living subject from oxidative damage, comprising: preparing the aquaporin water according to claim 1; and providing the aquaporin water to the living subject, wherein the aquaporin water is used for anti-oxidation to protect the cells of the living subject from oxidative damage.

15. A method for maintaining telomere length in cells of a living subject, comprising: preparing the aquaporin water according to claim 1; and providing the aquaporin water to the living subject, wherein the aquaporin water is used to maintain the telomere length of cells of the living subject.

16. A method for downregulating a gene associated with inflammation and aging in a living subject, comprising: preparing the aquaporin water according to claim 1; as well as The aquaporin water is provided to the living subject, wherein the aquaporin water is used for anti-inflammation to down-regulate the gene of the living subject.

17. A method for preparing functional water having enhanced cell penetration ability, comprising: Ceramics are prepared by melting natural clay and iron-containing sand; immersing the ceramic in water at a predetermined temperature and a predetermined ceramic-water weight ratio for a first predetermined time to obtain the functional water, and ultrasonically treating the ceramic in the water at the predetermined temperature and a predetermined ultrasonic power for a second predetermined time; and Applying the ultrasonically treated water to flow through a column filled with the ceramic at the predetermined temperature, thereby obtaining the functional water, wherein the water flows through the column at a predetermined water flow-ceramic weight ratio or at a third predetermined time required for a unit volume of water to pass through a unit weight of ceramic; The ceramic causes structural changes in at least some of the water molecules of the water.

18. The method according to claim 17, further comprising verifying the ability of the functional water by: placing cells of a living subject in the functional water as a culture medium; placing the cells of the living subject in deionized water as a control culture medium; and The cells in the culture medium and the control culture medium were measured using corresponding kits to determine the capacity of the functional water.

19. The method of claim 18, wherein the capacity of the aquaporin water comprises: increasing the viability of the cells; protecting said cells from oxidative damage; maintaining telomere length of the cells; as well as Down-regulating genes associated with inflammation and aging in the living subject.

20. The method of claim 18, further comprising: Before placing the cells in the functional water, placing a growth promoting substance in the functional water serving as the culture medium; as well as The culture medium is filtered to remove sediment and bacteria in the functional water.

21. The method of claim 18, wherein the growth promoting substance comprises: Minimum Essential Medium (MEM) as basal medium; As well as fetal bovine serum (FBS) and penicillin / streptavidin (P / S) as supplements.

22. The method of claim 17, wherein the ceramic is Tadanoumi ceramic, the functional water is aquaporin water, the predetermined ceramic-water weight ratio is a ceramic-water ratio of 0.01-0.1 g / ml, the predetermined temperature is room temperature, the first predetermined time period is 18-36 hours, the predetermined ultrasonic power is 100 watts, the second predetermined time period is 3-7.5 minutes, the predetermined water flow-ceramic weight ratio is 0.01 liters / minute / gram, and the third predetermined time required for 1 liter of water to pass through 1 gram of ceramic is 0.01 seconds.

23. Aquaporin water prepared by the method according to claim 17, wherein water molecules of the aquaporin water have a HOH bond angle α, and the HOH bond angle α1 of the altered water molecules is greater than the conventional HOH bond angle α0 of the water molecules of the water before treatment.

24. The aquaporin water according to claim 23, wherein the HOH bond angle α1 is about 120°, and the conventional bond angle α0 is 104.45°.

25. An aqueous solution having aquaporin water according to claim 23, wherein: The volume of the changed water molecules in the aqueous solution is V1; the volume of the unchanged water molecules in the aqueous solution is V0; the volume of the aqueous solution is V, V=V1+V0, and V1 / V0>1.

26. A method for enhancing the elimination of metabolic waste products in a living subject, comprising: preparing the functional water according to claim 17; and providing the functional water to the living subject, wherein the functional water is used to enhance the elimination of the metabolic waste from the cells of the living subject.

27. The method of claim 26, wherein the metabolic waste product comprises hydrogen peroxide.

28. An aqueous solution comprising: a number of first water molecules having a first volume V1 and a first average HOH bond angle α1 between two HO bonds in each of the first water molecules; as well as A number of second water molecules having a second volume V0 have a second average HOH bond angle α0 between two HO bonds in each of the second water molecules.

29. The aqueous solution of claim 28, wherein the first average HOH bond angle α1 is greater than the second average HOH bond angle α0.

30. The aqueous solution of claim 29, wherein the first average HOH bond angle α1 is 120°, and the second average HOH bond angle α0 is about 104.45°.

31. The aqueous solution according to claim 28, having a volume V, wherein V = V1 + V0, and wherein V1 / V0>1.

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