TW200423986A - Methods of treating disorders by altering ion flux across cell membranes with electric fields - Google Patents

Abstract

The invention relates to methods and devices for treating disorders with electric current or electric field therapy. The invention uses applied electric current or current induced by an external electric field to manipulate movement of ions across cell membranes and to alter ionic concentrations. The invention is useful, for example, for treating hyperproliferative and cardiovascular disorders and for ameliorating the effects of stress.

Description

200423986 发明. Description of the invention: I [No. Mingming's technical erosion field] I Cross-referenced related applications This application is an application filed on December 14, 2001 and announced on December 5, 2002 Part of US Application No. ι〇 / 〇ΐ7, 〇 05 continues the application. This application also benefits from US Provisional Application No. 60 / 433,766, which was filed on December 17, 2002, and US Provisional Application No. 60 / 399,249, which was filed on July 30, 2002. The entire contents of all the aforementioned applications are incorporated herein by reference. [Prior Art] Background of the Invention A wide variety of electrical therapy devices are known. Typically, the electrodes of a device are in contact with the patient, in which case the electrotherapy device uses an applied current and may be referred to as an electric cwrreni therapy device. Examples include TENS or PENS (Ghoname, E · A ·, et al ·, Anesth · Analg "88: 841-46 (1999); Lee, RC" et al ·, J Bum Care Rehabil "14: 319-335 (1993 )) 〇 Right pole; there is contact with the patient, the electrotherapy device uses an external electric field 20 (hereinafter referred to as "EF") to induce current in the patient, and can be called an electric field Or potential (eiectric household therapy). Ef generates surface charges in all conducting bodies (including animal or human bodies). When EF is applied, positive and negative charges will appear on the opposite sides of the body (opposite sides) When the electric field alternates, the electric charge will be transferred at the position 6 200423986, causing an alternating current in the body (see Hara, H., et al., Niigata Med., 75: 265-73 (1961)). In 1972 'Japan's Ministry of Health and Welfare has approved an electrical stimulation device (approval number 5 147〇〇ΒΒΖ0094). In 19 and 8 years, the USFDA approved electrical stimulation for the treatment of bone diseases. However, the treatment literature Reports a wide variety of biological aspects of electrical stimulation Reactions. For example, 'external sinusoidal alternating fields (ac EF) have been shown to alter cell type in other organisms, protein synthesis in fibroblasts, and redistribution of embedded membrane proteins. (redistribution), DNA synthesis in chondrocytes, intracellular # 5 ion concentration, microfilament structure in human hepatoma cells, and electrolyte levels in the blood (Kim, YV, et al., Bioelectromagnetics, 19: 366-376 (1998); Cho, MR ·, et al ·, FASEB J., 13: 677-682 (1999); Hara, H ·, Niigata Med ·, 15 75: 265-73 (1961 )). Some researchers believe that many of the observed effects are not directly attributed to EF, but are due to the fact that EF is in primary cellular structures such as membrane-receptor complexes and ion-converter channels. -transport channels). Although the biological effects of induced currents have been studied in the last 25 to 20 years, most of the research is due to people who are exposed to strong electric or magnetic fields from high transmission power lines and related electrical devices. Was inspired. For example, staff of utility companies are routinely exposed to an electric field of 50-500 kV / m and a magnetic field as high as 5 G, while the general public is usually exposed to an electric field of 1-10 kV / m and at most 2 G's magnetic field (Portier, 7 CJ & Wolfe, MS (eds.) Assessment of Health Effects from Exposure to Power-line Frequency Electric and Magnetic Fields, NIEHS Publ. No. 98-3981 (National Institute of Environmental Health Sciences, 1998 )). Previous techniques lack adequate research on the efficacy of relatively low voltages and weak electric fields. In addition, the traditional EF therapy uses a south voltage and does not account for the differences in EF intensity in different regions of the morphology across the body. In short, as pointed out by Sporer in U.S. Patent No. 5,387,231, "Prior art has not intended a combination of appropriate, effective electrical parameters for truly effective electrotherapy. Previous art devices have generally been at very high voltages. Or operate at very high currents, both of which can produce a diathermy effect in the tissue being treated. In many cases, prior art may mention one or the other of various electrical parameters , But carelessly considers the importance of other parameters. ”Because the first technique shows a completely different biological response, and relies on inaccurate measurement methods and focus on the efficacy of high voltage and high current, it still exists There is a need to identify specific parameters for use in electrotherapy, particularly electrotherapy using relatively low voltage and current. [Summary of the invention] Summary of the invention The inventors have determined parameter values of £ 17 and applied current that have successfully treated a specific disorder. These parameters include, for example, frequency (in Hertz), voltage (in volts), induced current density (in mA / m2), applied current density (in mA / m2), individual continuous 200423986 The duration of storm 2 (in minutes, hours, and days), and the total number of storms N · times (whether in one continuous exposure cycle or the sum of multiple consecutive exposure cycles). As used herein, "average, the applied current density and" average, the induced current density "means the generation of at least one organism of interest (for example, a human, animal, plant, or the like). —Parts, or their cells) the average current per unit area of the cell membrane. For example, if the biological system of interest is a human and the part of interest is the entire hand of the human, the average current density is the average value for the entire hand, that is, the average current density is 10 degrees as The sum of the current densities of the various parts of the hand divided by the sum of their areas. Special formulas and techniques described later here are used to estimate the average applied current density and average induced current density. Unless explicitly stated otherwise, the term “organism” encompasses both humans and other types of organisms. 15 A specific example of the present invention relies on an applied electric current. Preferably, the applied current density is in the range of about 10 to about 2000 lnA / m2. Another embodiment of the invention relies on a particularly low amount of induced current to control the movement of ions across the cell membrane. In order to treat a disorder caused or caused by an abnormal ion concentration of one of 20 in a cell of a living body, specific examples of the induced current include exposing the living body to an external electric field which generates a voltage of about 0.001 mA / m2 to about 15 mA / m2, preferably about 0.001 mA / m2 to about 10 mA / m2, more preferably about 0.01 mA / m2 to about 2 mA / m2, the average (mean) induced current density across the cell membrane. In a preferred embodiment, the field (E) of the external electric 9 200423986 is measured by a term in the formula E = Ι / εοωδ, where S is a section of an electric field measurement sensor. , Ε0 is an induction rate in a vacuum, I is a current, ω is 2πί, and f is a frequency. It is also preferable to measure the induced current (J) by the term in the formula J == I / B 5, where I is a measured current and B is a circle expressed as B = Α2 / 4π Area (circle area), A is a circle expressed as A = 2icr, and r is a radius. In an additional preferred embodiment of the invention, the induced current density is generated throughout the cell membrane for a continuous period of about 10 minutes to about 240 minutes. In 10 repeated implementations (In reapplication), the average induced current density is preferably generated over a continuous period of about 30 minutes to about 90 minutes, preferably resulting in a total exposure time of less than about 1,500 minutes. The two specific examples of applied current and induced current outside the present invention can be applied to an entire body or only a part of the body. The part of the body 15 may include a limb, an organ, defined body tissue, a part of the body (such as the trunk), a body system, or a subsection of the same. A trained individual can determine a particular obstacle to apply to an entire body or part of the body according to the present invention. The invention may further include providing a calcium supplement, a one-dimensional 20-vitamin D supplement, a plant agglutination (ectin) supplement, or a combination of these supplements to the organism. Preferably, the plant lectin supplement comprises Concanavalin A or wheat germ agglutinin. In a preferred embodiment, the present invention changes or affects about about 10 200423986 or other cations or polyvalent cations (including cationic electrolytes and proteins that play an important role in activating extracellular Ca-2 + -associated electro-sensitive calcium receptor (CaR) in extracellular fluid) Flow. 5 An alternative embodiment of the present invention relates to a device used as an EF therapy. A preferred EF therapy device is an electric field therapy device, which includes: a main electrode and a counter electrode (0pp0Sed electrode); a voltage for applying a voltage to the electrodes to generate p (voltage generator); an inductive current generator that controls an external electric field by changing the electrical voltage or the distance between the opposite electrode and the living body or part thereof; and a means for driving the voltage generator power supply. Preferably, the voltage generator has a booster coil and the voltage generator is grounded at a middle point or one end of the booster coil. In a more preferred treatment device of the present invention, it has a main 15 electrode and an opposite electrode, the opposite electrode is arranged close to a human body's head, shoulders, abdomen, waist or hips, And the distance between the opposite electrode and the surface of the torso range of the human individual is about 丨 to even cm, more preferably about 1 to 15 cm. In alternative aspects, the opposing electrode system is a ceiling, wall, floor, furniture, or other object or surface within the phase. 2〇 Alternative—An alternative specific example involves determining the optimal parameters for the needle shaft EF or external current therapy. -A better method to determine the best parameters for the occupational law includes the following steps: (i) identify a desired biological response to be induced in a living organism; (ii) screen or measure a The average 11 200423986 induced current density on the cell membrane of an organism or a tissue sample or cell derived from the organism; (iii) screening or measuring an external electric field that is separated from the organism, sample or A specific distance of the culture will produce a screened or measured induced current density; (iv) a screen or measurement of a continuous time period used to produce a screened or measured induced current density on the cell membranes; (v) Applying 5 the screened or measured electric field to the organism, sample or culture to produce the screened or measured induced current density on the cell membranes for a continuous period of time that is screened or measured; (vi) Determine the extent to which the desired biological response occurs; (vii) selectively repeat any of steps (ii) to (vi); and / or (viii) confirm that the desired biological response is best elicited Work 10 The induced current density values are measured or screened, as a value of the external electric field is measured or screened or as a value of the continuous time period is measured or screened. For this specific example, the term "measurement" includes situations where the experimenter chooses the parameter values unconsciously, intentionally, or initially. For example, the term "measurement" includes a case where an EF device produces a random or initially unknown 15 average induced current density, and the researcher directly or indirectly decides how much that amount. The invention is further illustrated by the following figures and detailed description. Brief Description of the Drawings 20 Figure 1 shows an electric field exposure jidl (field exposure dish) in an EF exposure system; Figure 2 shows the percentage of viable cells after EF exposure; Figure 3 shows the ratio of cells containing 12.5 There was a significant increase in the number of high [Ca2 +] j® cells in both EF exposed and unexposed cell suspensions of pg / ml Con-A 12 200423986. Figures 4A and 4B summarize the different concentrations of Con -A, results with EF-exposed cell cultures without ImM CaCl2; Figure 5 shows EF-exposed and unexposed cells containing phytohemaglutinin (5 PHA). In both cells, high [〇32 +]. There was a significant increase in cells; Figure 6 shows that when supplemented with 3.125-12.5 pg / ml of Con-A, both EF exposed and unexposed cells were compared to those stimulated at 0.025 pg / ml Con-A cells have a significant increase in high [Ca2 +] c ^ cells; Figure 7 illustrates the increase in calcium concentration induced by ConA in splenocyte cells; Figure 8 shows Time course of DiBAC staining intensity in BALB 3T3 mouse embryo cells stimulated with a final concentration of 0.4 μM A23187; Figure 9 shows an electric field that produces a current density of about 200 uA / cm2 at 100 Hz (EF) Effect on membrane potential in BALB3T3; Figure 10 also shows the effect of an electric field (EF) generating a current density of about 200 μA / (: ηι2 at 100 Hz on membrane potential in BALB 3T3; Figure 11 Shows the effect of stress on the level of plasma adrenocorticotropic hormone (hereinafter referred to as "ACTH"); Figures 12A and 12B show exposure to EF in normal and ovariectomized (B) rats Effect of Plasma ACTH Level; Figure 13 shows EF exposure Effect on blood tight ACTH levels in normal rats (n =: 6); 13 200423986 Figures 14A and 14B show that EF exposure is constrained-induced in normal palate and Indian nest resection (B) rats (B) restraint-induced) changes in plasma glucose levels; Figures 15A and 15B show EF exposure on restraint-induced plasma lactate sites in normal tadpoles and Indosectomy 5 (B) rats Figure 16 shows the effect of EF exposure on restraint-induced plasma pyruvate levels in ovariectomized rats; Figure 17 shows the effect of EF exposure on ovariectomy Effect of Constrained _ 10 Induced White Blood Cell (WBC) Counts in Rats; Figure 18 illustrates an example of an electric field generated by the use of an EF therapy device (in this case 'a BioniTron chair from the White Brother Health Science Association) A conceptual outline of Fig. 19 is a schematic diagram 15 of a preferred EF therapy device of the present invention; Figs. 20A and 20B show another preferred EF therapy device; Figs. 21A and 21B show another A better EF therapy pack Figure 22 is a diagram showing a preferred electronic configuration of an EF therapy device. Figure 23A is a front view of a stimulated human body, and Figure 23B is a perspective view. Fig. 23C is a diagram showing an EF measurement sensor attached to the neck of the body; Fig. 24 shows a device for measuring an induced current generated by the EF therapy device Figure 14 shows the relationship between an applied voltage (appHed v〇itage) and an induced current; Figure 26 shows the relationship between the position of a head electrode and the current induced in the neck Relationship; 5, > _ Figure 27 illustrates the induced current density (mA / m2) at different locations of an ungrounded human individual; and Figure 28 shows the mitigation effect of EF exposure on different human symptoms. Embodiment 3 Detailed description of the preferred embodiment A. Method of regulating ion flow across the cell membrane (MethOd 0f Mofdulating Ion Flux Across Cell Membranes) An ion imbalance may originate from a disorder or condition, Or it could be a side effect of a medical treatment or supplement. The present invention changes the ion flow across the cell membrane by generating a current across the cell membrane. The invention also affects the composition of 15 cell membranes (such as its transmembrane proteins). The present invention can restore or balance cellular ionic homeostasis or change the membrane potential of cell membranes. Therefore, the present invention is applicable to prevent or treat disorders related to cell or extracellular ion concentrations such as calcium (Ca2), magnesium, sodium (Na +), potassium (κ +), and chlorine (cr) concentrations. 20 In order to treat disorders related to serum calcium concentration, the average induced current density generated across the cell membrane is preferably about 0.3 mA / m2 to about 0.6 mA / m2 'more preferably about 0.4 mA / m2 to about 0.5 mA / m2, the best is about 0.42 mA / m2. Using an externally applied current to treat a disorder related to serum maternal concentration 'The average externally applied current density is preferably about 60 mA / m2 to about 2,000,00,2004,23,986 mA / m2, and the average externally applied current density is A continuous period of time ranging from about 1 minute to about 20 minutes, more preferably from about 2 to about 10 minutes, is generated throughout the cell membrane. Tissues that can be administered by the methods of the present invention include, for example, musculo-skeletal tissues, tissues of the central and peripheral nervous system 5 (tissues of the central and peripheral nervous system), gastrointestinal system tissues, Reproductive system tissue (both male and female), pulmonary system tissues, cardiovascular system tissues, endocrine system tissues, immune system tissue (immune 10 system tissues), Lymphatic system tissues, and urogenital system tissues. Biofilms of eukaryotic cells, such as plasma membranes, are selectively permeable to these ion systems. Selective permeability allows to establish a membrane potential across the membrane. Cells use this membrane potential to transport molecules across the membrane. Many ions involved in generating a membrane potential perform vital functions. For example, a threshold concentration of I-bow ions in muscle cells initiates contraction. A threshold concentration of ma ion in exocrine cells of the pancreatic system triggers the secretion of digestive enzymes. Similarly, different concentrations of sodium and potassium ions are necessary for the electrical conduction of electrical impulses through nerve axons. A protein family widely known as voltage-gated ion channels maintains ion concentration and membrane potential. The voltage-gated ion channel is a transmembrane protein that contains ion-selective pores. The ion selects 16 200423986, and selects pores / syntheses depending on the configuration state of the distant channel (confomatial state) and allows ions. Through biofilm. The configuration state of the channel is affected by a voltage-sensitive portion that contains a charged (j amino acids) that reacts to the membrane potential. 5 The channel is Conductive (on / activated) or non-conductive (off / non-activated). Since special ions (ie, Ca2 +) are related to cardiovascular health, the present invention can be used to prevent and treat cardiovascular disorders. These Disorders include, for example, cardiomyopathy, dilated 10 congestive cardiomyopathy, hypertrophic cardiomyopathy, angina, variant angina, instability Unstable angina, atherosclerosis, aneurysms, abdominal aortic aneurysms, peripheral 15 arterial disease, blood pressure disorders (such as hypotension and Hypertension), orthostatic hypotension, chronic pericardiitis tis), arrhythmias, atrial fibrillation and flutter, heart disease, left ventricular hypertrophy, right 20 ventricular hypertrophy, Tachycardia, atrial tachycardia, ventricular tachycardia, and hypertension. The present invention can also be used to prevent or treat blood disorders. These blood disorders include, However, it is not limited to hyponatremia, hypernatremia 17 200423986 (hypernatremia), hypokalemi, hyperkalemia, hypocalcemia, and hypercalcemia. Hypercalcemia, hypophosphatemia, hyperphosphatemia, hypomagnesemia 5 and hypermagnesemia, and disorders of blood glucose regulation (such as diabetes , Adult diabetes, and adolescent diabetes). In a specific example of the present invention, a lectin and EF are co-applied to enhance Ca2 + flow across the cell membrane. Plant lectins useful in the present invention include, for example, concanavalin 10 (Akoncanavalin A) and wheat germ agglutinin. In another specific example, the ion current caused by the present invention is generated simultaneously with a calcium supplement. In another specific example, the ion current caused by the present invention is generated simultaneously with a vitamin D supplement or with a calcium supplement and a vitamin d supplement. Shuming's vitamin 0 supplements include, for example, Example 5 (Vitamin A (calciferol) and vitaminized alcohol). Likewise, the method of the present invention can be performed in conjunction with a supplemental light source that is applied to a biological sample or the surface of a patient. ㈣-a wavelength in the range of about 225 nm to about 700 nm. In a specific aspect of the present invention, the method and the method of the present invention are used to collectively engrave the light source 20 to emit a wavelength in the range of about 230 nm to about 313 nm. In an additional embodiment of the present invention, another molecule can be transferred across a cell membrane simultaneously with an ion current generated by the present invention. The additional molecules that can be transferred simultaneously with the ion current can be produced naturally by the body, or alternatively can be provided by means of supplementation (e.g., via a vitamin, etc.). Example 18 200423986 For example, cellular glucose uptake can be enhanced by the flow of calcium ions across a cell membrane. Additional molecules that can be transferred across a cell membrane simultaneously with an ion stream generated by the present invention include nutritional medicines (e.g., a nutritional supplement designed and formulated to help prevent or treat a disorder and / or condition). In addition, the method of the present invention can be used in conjunction with hyperalimeiitation treatment (e.g., administering nutrients beyond normal requirements to treat disorders, such as coma or severe burns or gastrointestinal disorders). Example 1 60 Hz Upregulates the cytoplasmic calcium (Ca2 +) level in spleen cells of mouse lectins stimulated by lectin 10 The EF exposure system used for this experiment consists of four parts: An electric field exposure dish made of polycarbonate; the function generator (SG-4101, IWATSU Co. Ltd., Tokyo, Japan); digital multi-meter ) (VOAC-7411 IWATSU, Tokyo, Japan); and control 15 devices (Hakuju Co. Ltd. Tokyo, Japan). Figure 1 shows an electric field exposure dish in an EF exposure system. The electric field exposure is covered by a cover (lid), a dish, and a doughnut-shaped insert (inner diameter: 12mm). An EF between two circular platinum electrodes (cell culture space) passes through the function generator Is generated and carefully adjusted by using the 20 controller and the digital multimeter. The field strength of the 60 Hz electric field is measured by measuring the cell culture space in the electric field exposure dish. Current density The current density is calculated by the formula: current density = 1 / S, where "Γ is the supplied cmrent (pA) and S is the cell 19 200423986 culture space (0 · 36π). Therefore, the current density can be calculated by: current density = 0.885Ι [μΑ / αη2]. Before EF exposure, about 1.5 ml of analysis buffer (137 mM NaCl, 5 mM KC1, 1 mM Na2HP〇4 5 mM glucose, 1 mM 5 CaCl2, 0.5 mM MgCl2, 0.1% (w / v) BSA and 10 mM HEPES pH 7.4) were poured into the electrode chamber. To avoid contact with cells and lower electrodes, a polycarbonate membrane (Isopore, MILLIPORE, MA USA) was placed between the jbi and the insert. Approximately 1 ml of the cell suspension was poured into a well / spacer and covered with a lid. 10 Cell preparation Female BALB / c mice (4-7 weeks old, available from CLEA Corporation (Tokyo, Japan)) housed in a traditional animal room equipped with a clean air filter The spleen was splenectomized and a cell suspension of spleen cells was prepared. To test cell viability (ceu 15 viability), the cells were cultured in Dulbecco's modified Eagle's medium supplemented with 10% fetal bovine serum (FSB). (SIGMA, MO, USA)). These cells were maintained in Hanks' Balanced Salt Solution (HBSS) (SIGMA, MO, USA) during the [Ca2 +] d test. The [Ca2 +] test was detected within 4 hours after the cell preparation. get on. Cells were stored at 4ac before use. The viability of EF-exposed cells was measured. Mouse spleen cells (5 X 106 cells / ml) were exposed to 60 Hz, 6 μA / cm2 or 60 μΑ / οηι2 EF at 37 ° C, 5% Co2 lasted 30 minutes and 24 hours. Sham cells were left in an electric field exposure dish for 30 minutes and 20 200423986 for 24 hours, but were not exposed to EF. At the end of the 30-minute and 24-hour exposures, the cell suspension collected from the electric field exposure was stained with 2.5 pg / ml propidium iodide at 4 ° C for 30 minutes, and the percentage of dead cells It is calculated by flow cytometry5. Cell preparation and analysis of lectins for analysis of high [Ca2 +] Jm cells Using spleen cells (106 cells / ml) at 37 ° C to contain 2.5 μM of fluoro-3-acetoxymethyl (fluo-3- HBSS of acetoxylmethyl) (Molecular Probes, USA) [Vandenberghe et al., 1990] was cultured for 20 minutes. The 10 cell suspension was then diluted 5 times with HBSS containing 1% FBS, incubated at 37 ° for 40 minutes, washed 3 times with analysis buffer, and the cells were suspended in the suspension. The analysis buffer became a concentration of 1x 106 / ml. The cell suspension was gently mixed throughout the cell preparation. 15 Considering the EMF and mitogen (Walleczek and

Liburdy (1990) reported a synergistic interaction between concanavalin A (Con-A) (Seikagaku Co., Tokyo, Japan) and phytohemagglutinin (Pha) (SIgmA, MO , USA). The experimental design used to determine 60 Hz (6 μA / cm2) EF for producing high [Ca2Um cells20 effect was designed taking into account the early analysis of the viability test results of the spleen cells of exposed mice 'We chose to use the best culture The following five experiments were performed with exposure conditions (60 Hz, 6 pA / cm2EF): ⑴ Cells suspended in HEPES-buffered solution (BS) + 1 mM CaCl2 21 200423986 were exposed to EF for a total of 40 minutes, and in the exposed Con-A was added at 12.5 pg / ml after the first 8 minutes. The control group consisted of EF-exposed cells with Con-A and EF-exposed cells without Con-A. High [Ca2 +] c-cell percentages were examined at determined exposure time points; 5 (2) Cells in HEPES-BS + 1 mM CaCl2 were exposed for a total of 12 minutes, and different concentrations (1 ng- 12.5 pg / ml Con_A is added after the first 4 minutes of exposure. The control group was essentially the same as the experimental group but without EF exposure; (3) Cells in HEPES-BS + 1 mM CaCl2 were exposed for a total of 10 8 minutes, and 5 pg / ml of PHA during the first 4 minutes of exposure Added later. The control group consisted of cells not exposed to EF with PHA and cells exposed to EF without PHA; (4) Cells suspended in HEPES-BS without CaCl2 were exposed for a total of 12 minutes and at different concentrations ( Con-A (1 ng-5 pg / ml) was added after the first 4 15 minutes of exposure. The control group was essentially the same as the experimental group but without EF exposure; and (5) In order to evaluate the sustained effect of EF exposure, cells suspended in HEPES-BS + 1 ml CaCl2 were exposed for a total of 4 minutes, and then at different concentrations (0.025 -12.5 pg / ml) of Con-A was added, and the next 8 minutes and 20 minutes without EF exposure were high [Ca ^ +]. The production of cells was followed by flow cytometry. The control group was essentially the same as the experimental group but without any EF exposure. Statistical Analysis Statistical analysis of cell viability was determined using the Student's t test. The data on the utility of EF exposure between groups 22 200423986 [Ca] c were analyzed by ANOVA (Analysis Of VAriance between groups), Stuart's t test, and paired t test. . All calculations for statistical analysis were performed in the MS-EXCEL® Japanese version (Microsoft Office software: Ver. 9.0.1 5 Microsoft Japan Inc. Tokyo, Japan). Results Figure 2 shows the percentage of viable cells after EF exposure. In all two replicates', either after exposure to 6 μA / cm2 or 60 μA / cm2, more than 98% of the cell lines were viable. 10 The number of high [Ca2 +] c cells increased significantly in both EF-exposed and unexposed cell suspensions containing 12.5 Mg / ml Con-A (Figure 3). In Fig. 3, the circle indicates a suspension without Con-A, the triangle indicates a suspension with Con-A exposed to EF, and the square indicates a suspension with Con-A not exposed to EF. High [Ca2 +] c cells in cell suspensions that did not survive the exposure were essentially unchanged. The reaction induced by cOnA was immediately noticed and the mitogen ( Mitogen) reached a saturation point within 5-8 minutes. The difference between exposed and unexposed Con-A-induced cells was insignificant (P > 0.05). 20 Figures 4A and 4B outline the different concentrations of Con-A, with and without

ImM CaCl2 results from EF-exposed cell cultures. Figure 4A shows the results of a culture without CaCl2 containing lmM. In Figure 4A, the cultures exposed by ef (black bars) and those not exposed to EF (white bars) contain ImM CaCh and different concentrations of con-A (0.01 23 200423986 pg / ml to 5 pg / ml). In the presence of CaCl2 (Figure 4A), EF significantly enhanced the Con-A-dependent [Ca2 +] c (/ > < 0.01: ANOVA). Although 0.675 -5.0 pg / ml (: 〇11-eight stimulated group is high [〇32 +] (: cell increase line is more substantial, but only 1.25 pg / ml and 2.5 pg / ml Con-A induction Significant difference was shown in 5 of the cells (cadaver <0.05; paired t-test). In Figure 4B, cultures exposed by EF (black bars) were compared to controls not exposed to EF. Group cultures (white bars) contained different concentrations of Con-A, but did not contain CaCl2. In the two groups of control group and EF exposure, in the state of cells without Ca2 + (Figure 4B) Con-A-dependent [Ca2 +] c rise is minimal 10. To determine whether EF-dependent [Ca2 +] c upregulation is restricted to Con-A, cells stimulated with PHA were also analyzed. Both EF-exposed and unexposed PHA-containing cells showed a significant increase in high [Ca2 +] c cells (Figure 5). However, compared to the non-exposed group, exposure to EF The increase in the cells is a significant OP < 0.05: paired t-test).

Compared to those cells stimulated with 0.025 pg / ml Con-A, add 3.125-12.5 pg / ml Con-A to the cell suspension (whether unexposed or initially exposed to EF for 4 minutes) Medium shows a significant increase on high [Ca2 +] c cells (Figure 6). Cells stimulated with 3.125 and 6.25 pg / ml Con-A exhibited a sustained increase in high [Ca2 +] c cells (which were flat about 8 minutes after Con-A stimulation), whereas they were stimulated at higher concentrations Con-A (12.5 pg / ml) cell cultures showed a decline on high [Ca2 +] j® cells approximately 4 minutes after Con-A stimulation. The enhanced effect of EF exposure 24 200423986 was significantly verifiable only in the presence of 6.25 pg / ml Con-A at 2-4 minutes (P < 0.05: paired t-test). Example 2-Effect of Low Frequency Electric Field on Vasoactive 5 Substance-Induced Intracellular Calcium (Ca2 +) Response in Human Vascular Endothelial Cells In order to evaluate the effect of EF on human vascular endothelial cells (hereinafter referred to as HUVEC), intracellular calcium levels were detected by HUVEC stimulated by ATP and histamine. To evaluate the effectiveness of EF on HUVEC, HUVEC was exposed to a 50 Hz (30,000 V / m), 3,000 volt EF. The EF induced current density on HUVEC 10 is estimated to be 0.42 mA / m2. HUVEC was exposed to these test parameters for 24 hours. After exposure, the cytoplasmic free Ca2 + concentration was determined by a fluo3 flow cytometer. A change in fluo3 image intensity was confirmed by real-time exposure to a real-exposure confocal laser microscopy (15). The results confirmed that EF increased the calcium concentration in HUVEC. B. The method of treating proliferative cell disorders is to treat proliferative cell disorders, especially those involving differentiated fibroblast cells. The average induced current density generated across the cell membrane is preferably It is about 0.1 mA / m2 to about 2 20 mA / m2, more preferably about 0.2 mA / m2 to about 1.2 mA / m2, and still more preferably about 0.29 mA / m2 to about 1.12 mA / m2. With an applied current, the average applied current density generated across the cell membrane is preferably about 10 mA / m2 to about 100 mA / m2. The fibroblast cell line is an embryonic mesoderm tissue 25 200423986 (embryonic mesoderm tissue ) Cell type. Fibroblasts can be cultured in vitro and secrete matrix proteins such as laminin, fibronectin, and collagen. Cultured fibroblasts usually do not differentiate like tissue fibroblasts. However, with appropriate stimulation, fibroblasts have the ability to differentiate into many cell types such as, for example, adipocytes, connective tissue cells, muscle cells, collagen fibers, and the like. Specific fibroblasts are able to differentiate into many cell types related to connective tissue and the musculoskeletal system. Methods to control the growth of undifferentiated fibroblasts in vivo or in vitro can be applied to control derived from fibroblasts The growth of differentiated cells. For example, hyperproliferative disorders of the musculoskeletal system can be controlled or prevented by preventing fibroblast growth. We determined that an applied current of about 10, 50, or 100 mA / m2 was generated across the cell membrane at a density of about 24 hours / day for at least about 7 days to inhibit the growth of fibroblasts in a current density-dependent manner. Growth 0 Hyperproliferative disorders include, for example, neoplasms associated with connective and musculoskeletal tissues, such as fibrosarcoma 20 (rhabdomyosarcoma), myxosarcoma, chondrosarcoma, Osteogenic sarcoma, chordoma, and liposarcoma. Additional hyperproliferative disorders that can be prevented, ameliorated or treated using the methods of the present invention include, for example, the progression and / or metastasis of malignancies 26 200423986 (malignancies). These malignancies include those located in, for example, the abdomen ( the abdomen), bones, brain, breasts, colon, digestive system, endocrine glands (adrenal, parathyroid, pituitary, testis, ovary, thymus, thyroid), eyes, head and neck, liver, 5 lymphatic system, Neoplasms of the nervous system (central and peripheral), pancreas, pelvis, peritoneum, skin, soft tissue, spleen, chest, and urogenital tract, leukemias [including acute Anterior bone somatic (acute promyelocytic), acute lymphocytic leukemia, acute bone marrow: acute myelocytic leukemia, myeloblastic, anterior bone marrow: promyelocytic ), Bone marrow mononuclear (myelomonocytic), monocytic (monocytic), red blood cells Erythroleukemia], lymphomas [including Hodgkins and non-Hodgkins 15 lymphomas], multiple myeloma: multiple myeloma, colon cancer (colon carcinoma, prostate cancer, lung cancer, small cell lung cancer, bronchogenic carcinoma, testicular cancer, cervical cancer, Yin Chao Ovarian cancer, breast 20 cancer, angiosarcoma, lymphangiosarcoma, endotheliosarcoma, lymphangioendotheliosarcoma, synovial membrane cancer (synovioma), Mesothelioma, Ewing's sarcoma, leiomyosarcoma, squamous cell carcinoma 27 200423986 (squamous cell carcinoma), basal cell carcinoma, pancreatic cancer ), Renal cell carcinoma, Wilm's tumor, liver cancer (hepatoma), bile duct carcinoma, adenocarcinoma 5 (epithelial carcinoma), melanoma, sweat gland carcinoma, sebaceous gland carcinoma, breast Papillary carcinoma, papillary adenocarcinoma, glioma, astrocytoma, meduloblastoma, craniopharyngioma, craniopharyngioma, Ependymoma, pinealoma, hemangioblastoma, acoustic neuroma, oligodendroglioma, meiangioma, neuroblastoma (Neuroblastoma), retinoblastoma, 15 bladder cancer, embryo cancer (embryoTial carcinoma), cystadenocarcinoma, medullary carcinoma, choriocarcinoma ), And seminoma. Example 3-EF exposure to mouse splenocytes and 3T3 / A31 fibers Effect of 20 Ca2 + Concentration in Uteroblasts on Mouse Spleen Cells In order to determine the effect of EF on calcium ion concentration in mouse spleen cells, a special EF electric field exposure at 60 Hz was applied to mouse spleen cells. Mice were excised from the spleen under anesthesia. In a 60 mm dish, the spleen was injected with pbs (a phosphate buffer solution containing 0.083% NhCl 28 200423986). The cells were resuspended and maintained in Hanks' Balanced Salt Solution (HBSS) (SIGMA, MO, USA) during [Ca2 +] c assay, which, 2+]. Detection was performed within 4 hours after cell preparation. Cells were stored at 4 ° C before use. 5 Application of a 60 Hz EF to spleen cells produced 6, 20, 60, and 200 μA / cm2 of applied current density. Splenocytes were exposed to these conditions for 4 minutes, and after these exposures, the splenocyte samples were stimulated with concanavalin A (ConA). After spleen cells were stimulated with cOnA, the cytoplasmic free Ca2 + concentration was measured by a fluo3 flow cytometer. 10 This experiment confirms that ConA increases the approximate concentration in splenocytes. The ma ion concentration was increased by the application of a 6-200 μA / cm2 EF. More importantly, the increase in dance ion concentration depends on the current density (see Figure 7 where the Y-axis shows the calcium concentration and the X-axis shows the time in minutes). Effect on BALB 3T3 15 In order to determine the effect of EF on calcium ion concentration in mouse 3T3 / A31 fibroblasts, these 3T3 cells were directed to an EF at 60 Hz. The 3T3 cell line was obtained from the tlle cell bank of the Japanese National Research Center for

Protozoan Disease) and grown at 37 ° C in DMEM containing 5% FCS and 10 mM 20 HEPES. EF produced a 200 μA / cm2 applied current density throughout the cells. After 2 minutes of exposure, the cytoplasmic free Ca2 + concentration was measured by a fluO3 flow cytometer, and the fluo3 flow cytometer showed an increase in calcium concentration in these cells. A change in the fluo3 image intensity was confirmed with a conjugate focus laser microscope 29 200423986. Case 4-Effects of Calcium Channels (Ionophore) and EF on the Membrane Repackaging in BALB 3T3

Figure 8 shows that the membrane potential of mouse BALB 3T3 / A31 fibers 5 embryonic / embryonic cells is altered by about ionophore. Figure 8 shows the time course of DiBAC intensity in BALB 3T3 cells stimulated at a final concentration of 0.4 mM A23187. A23187 is a monounsaturated acid (11101100 & 1 ±) (^ 7valence3 (: 丨 (1)) extracted from fenre / plus / wyees c / wrir sign / Ls / s. DiBAC is a fluorescent stain, which enters the cell membrane when the potential of the cell membrane changes. Therefore, when the cell membrane of these BALB 3T3 cells is depolarized, DiBAC enters those cell membranes, thereby increasing the DiBAC signal intensity (Y-axis) in these BALB 3T3 cells.

Figure 9 shows the effect of an electric field (EF) at 100 Hz on the membrane potential in BALB 3T3, which produces a current density of about 200 mA / cm2 at 15 degrees. Changes in membrane potential were measured by flow cytometry. The methodology of flow cytometry is shown below. Cultures in DMEM were supplemented with 5% FCS 10 mM HEPES. It was then de-touched with 0.02% trypsin and 0.025% EDTA. It was then resuspended in HEPES buffer solution (137 mM NaCl, 5 mM KC1, 1 mM 20 Na2HP04, 5 mM glucose, 1 mM CaC12, 0.5 mM MgC12, 0.1% (w / v) BSA, and 10 mM HEPES pH 7.4 )in. It was then loaded with DiBAC4 (3) at a final concentration of 200 nM. It was cultivated at 37T: for> 5 minutes. Next, flow cytometry is performed. Figure 10 also shows the effect of an electric field (EF) at 100 Hz on the membrane potential in BALB 3T3 30 200423986. The electric field produces a current density of about 200 mA / cm2. Implementation of changes in synovial fibroblasts (Synovia! ^ Gap Junction Intercellular 5 Commimication). ― We examined the effects of low-level currents on intercellular junctions (GJIC) regulated by the connexin43 protein. Synovial fibroblasts The confluent single cell layer of cells (HIG-82) and neuroblastoma cells (5Y) was exposed to each in a bath solution, 0-75mA / m2 (0-56 10 mV / m '60 Hz) 'and single-channel conductance (S) (ngie_channei conductance), cell membrane current-volt (IV) curve, and Ca2 + influx were measured using nystatin double- and single_patch methods In the cells exposed to 20 mA / m2 (at 0.76ρΑ and 0.39 pA, respectively), the conductance of the closed and open states of the gap junction channel (gap_juncti〇n channei) in HIG-82 cells were each significantly changed Decreased; no utility occurs in the conductance of the gap junction channels between 5Y cells. The low current density as low as 10 mA / m2 significantly increases Ca2 + influx in HIGH-82 cells, but is not effective for 5Y cells. This Two types of fine The IV curve of the plasma membrane (plaSma membrane) does not depend on the current of 60-Hz, 0-75 mA / m2, indicating that the effect of 60-Hz current 20 on GJIC in HIG_82 cells is not affected by a change in membrane potential. The conclusion is that low-level extracellular currents can change GJIC in synovial cells via a mechanism that does not depend on membrane potential changes, but may depend on the mechanism of Ca2 + influx. The results indicate that GJIC-regulated responses in synovial cells (For example, 31 200423986 their secretory response to pro-inflammatory cytokines) can be antagonized by the application of extracellular low-frequency currents. C. Method of Reducing Stress The invention can be applied to prevent or treat stress and disorders related to stress, such as reduced immune-system function, infection, hypertension, atherosclerosis, and insulin-resistant blood Insulin-resistance-dyslipidemia syndrome. For the treatment of stress, immunosuppressive disorders, and to reduce the level of ACTH or cortisol, the average induced current density generated across the cell membrane is preferably about 0.03 mA / m2 to about 12 mA / m2, more preferably 0.035 mA / m2 to about 11.1 mA / m2. With the applied current, the average applied current density is preferably about 60 mA / m2 to about 600 mA / m2. Stress is associated with many health disorders, including hypertension, atherosclerosis, and signs of insulin-resistant dyslipidemia, as well as established immune dysfunction (VanitallieT.B., Meiah // Tear, 51: 40- 5 (2002)). Researchers have found that stress can affect the homeostasis of adrenocortical hormones, such as cortisol and corticosterone. The hormone corticosterone is produced by the adrenal glands, and its changes are a common indicator of stress. In a report involving mice exposed to electric fields of up to 50 kV / m and 60 Hz, the decrease in plasma corticosterone concentration was observed 'but only in the early part of the exposure period (Hackman, RM & Graves, HB " 5e / mv · iVewra / B / o / · 32: 201-213 (1981)). Similarly, Portet and Cabanes reported that when rabbits and rats were exposed to 32 200423986 50 kV / m, 50 Hz, the lower Cortisol level was found in the adrenal glands, but cortisol concentration in the blood was not found (Portet, R. & Cabanes, J., 9: 95-104 (1988)). ACTH is a kind of The peptide of the pituitary gland shows a 5-peptide, and almost exclusively controls the secretion of cortisol. The ACTH level is a powerful indicator of body pressure in terms of physical function, mainly because ACTH acts on Controls the secretion of cortisol, a major anti-inflammatory molecule that is extremely important for stress responses such as traumatic events. Interestingly, researchers have found no increase in ACTH level 10 after 30-120 days of electric field exposure ( Free, MJ ·, et al ·, Bioelectro magnetics 2: 105-m (1981)). In one study, when rats were exposed to 100 kV / m, 60 Hz, for 1-3 hours, no change in plasma ACTH was found (Quinlan, WJ "et al ”Bioelectromagnetics 6: 381-389 (1985)). When mice were exposed to 10 kV / m at 50 Hz, serum ACTH concentrations were 15 degrees higher than those in the control group (deBmyn, L. & deJager , L., Environ. Res. 65: 149-160 (1994)). Lipid staining is increased in an adrenal cortex, but only in males. The authors conclude that the electric field is a stressor. The changed Blood ACTH concentrations were also found in rats exposed to a 15 kV / m, 60 Hz electric field for 30 days (Marino, 20 Α · Α ·, et al ·, Physiol · Chem · Phys. 9: 433-441 (1977)) Conversely, we have confirmed that applying an electric field at a particular parameter to a test animal results in a decrease in stress-induced ACTH concentration. For example, applying an electric field of 17,500 V / m (50 Hz) , A voltage of 7,000 V, and an induced current density of about 0.035-0.5 mA / m2, which lasted 33 200423986 for a period of 60 minutes Resulting in lower test animals stress-induced serum level of ACTH. Effects of Two 50 H_z Electric Fields on Plasma ACTH,% Glucose, Lactam, and Pyruvate in Rats Who Are Absent 5 Electric Field Exposure System The EF exposure system used in this example consists of the following three main parts Composition: a high voltage generator (Healthtron TM, maximum output voltage: 9,000 V; Hakuju Institute for Health Science Co. Ltd. Tokyo, Japan), a constant voltage power supply (TOKYO SEIDEN, 10 Tokyo, Japan), and EF exposure cages (cages). These exposure cages are made of a cylindrical plastic cage (φ: 400 mm, height · 400 mm) and two made of stainless steel above and below the cylindrical cage. Electrode (1,200 x 1,200 mm). In order to form EF (50 Hz; 17,500 V / m) in the cage, a stable alternating current (50 Hz; 7,000 V) was applied to the upper electrode 15 female experimental animals, 7 weeks old, Wistar rats weighing 300-350 g were purchased from Charles River Japan, Inc. (Tokyo, Japan) and housed in a traditional animal room equipped with an air purification filter Restraint Stress 20 Rats Borrow Each package was constrained with a thin polycarbonate plate and placed on the lower electrode for 30 minutes. Experimental design The effect of EF on the restraint pressure was determined as described below. For evaluation Using the thin polycarbonate plate restraint method, 6 rats were divided into two groups: 34 200423986: only restraint and restraint plus diazepine treatment. To detect the effects of exposure to EF, we used normal and Ovariectomized rats. Normal rats are divided into two groups: restraint and constraint plus EF. In addition, ovariectomized rats are also divided into the following four subgroups: simulated (A1), simulated EF exposure plus restraint (A2), EF exposure plus restraint (A3), simulated EF exposure plus diazepine treatment and restraint (A4). Ovariectomy was performed 4 weeks before the experiment Executed. The EF exposure and restraint treatment applied in this study is shown below: Rats were exposed to% Η, 17,500 V / m EF for a total of 1 hour. Rats were treated with thin polycarbonate plates Subject to 10 constraints for the second half of the EF exposure period The experimental design of the control group was the same as that of the experimental group, except that there was no EF exposure. Collection of Enterprise Liquid Samples 1 ml of blood was collected from the subclavian vein before the experiment was started, and the plasma was collected at 40%. c. It was prepared by centrifugation at I, 5000 g for I5. Plasma was stored at -80 before hormone measurement. c. After the experiment, 3 ml of whole blood from each rat was collected into a glass tube containing 9 mg eDTA by cardiac pimcture under anesthesia. 1 ml of blood is used to analyze the blood condition. The other 2 mi was centrifuged (at 4 ° C, min at 20 1,5⑻X g) and the supernatant was stored at -80. C until hormones, glucose, lactic acid, and pyruvate are measured. Hematology analysis including red and white blood cell counts, platelet counts, hematocrit, and heme levels is performed using an automated 35 200423986 automatic multi-hemocytometer (Sysmec CC-78, Sysmec inc., Tokyo, Japan). Blood polydextrose, lactic acid, and pyruvate levels were measured with an automatic analyzer (7170 Hitachi, Hitachi Co. Ltd., Tokyo, Japan). The ACTH level uses an ACTH radioimmunoassay kit (ACTH IRMA, MITSUBISHI CHEMICAL Co. Ltd.) and a gamma counter (Auto-Gamma 5530 Gamma Counting System, Packard Instrument Co. ltd. ) To measure. Plasma corticosterone levels were measured using a commercial kit (ImmuChem Double Antibody 10 Corticosterone kit, ICN Biomedicals Inc.). Statistical analysis results are expressed as mean ± standard error of the mean (S.E.) or data sets such as median, 25th percentile, 75th percentile, minimum, and maximum. Statistical I5 significance of difference between paired groups was calculated by the Stuart's t test, and significance was defined as P < 0.05. All calculations for statistical analysis were performed in the MS-EXCEL® Japanese version (Microsoft Office software. Ver. 9.0.1, Microsoft Japan Inc. Tokyo, Japan). 20 Results Changes in plasma level induced by restraint stress. Figure 11 shows the effect of stress on plasma ACTH level. Rats were administered intraperitoneally with 1 mg / kg B.W. diazepam (filled circle 36 200423986 shape) or saline (open square). Thirty minutes after the diazepine administration was performed, the rats were restrained to provoke a stress response. Figure 11 shows the ACTH levels of individual rats 30 minutes after the start of restraint. In this constraint-only group, the values before and after the constraint period (mean ± SE) are 231 5 ± 135 and 1177 ± 325 pg / ml, while in the constraint plus diazepine group, It is 358 ± 73 and 810 ± 121 pg / ml. Comparing the ACTH levels before and after the restraint pressure in each group, the 30-minute restraint increased the plasma ACTH level by 5.1 times and 2.3 times in the only restraint and the restraint + diazepine groups, respectively. Effects of EF Exposure on Constrained Induced Plasma ACT1 ^ ± Figures 12A and 12B show the effects of EF exposure on plasma acth levels in normal (A) and ovariectomized (B) rats. All rats were restrained during the second half of the EF storm. In the following groups, the Kimchi ACTH level was measured 60 minutes before and after EF exposure: no treatment (n = 6), only 15 constraints (simulated, n = 6), constraints at EF (EF , N = 6) and constraints and diazepine in simulated EF (simulated and diazepine, n = 6). The addition of diazepine occurred 30 minutes before the beginning of the EF period. The data is represented as four boxes, showing the horizontal line that divides each main four box into two small four boxes representing the median, and the horizontal line that forms the bottom side of each main four box represents 20 μ, quantiles, the horizontal line forming the top side of each major quadrilateral represents the 75th percentile, shown as the horizontal line above each major quadrilateral represents the maximum, and is displayed as the major quadrilateral The horizontal line below the box represents the minimum value. Pre values are not displayed. *: p < 0.05 is from 37 200423986 value before constraint. f: P < 0.05 is from the untreated group. In ovariectomized rats, the plasma ACTH level of the unconstrained group did not show any change during the 60 minute period. In the other three groups, the ACTH level was raised during the restraint period (Figure 12β). Comparisons were made between the first 5 periods and after the period. Plasma levels were increased by 186, 134, and 137 times in the "constraints only", "constraints and EF," and "restraint and diazepine" groups, respectively. Figure 13 shows the effect of EF exposure on blood polyacth levels in normal rats (n = 6). Data are expressed as a median, 25th percentile, 75th 10th percentile, minimum, and maximum. Figures 12A and 13 show changes in plasma levels of ACTH and corticosterone in normal rats. In this "only constraint," and "" constraint and EF, the ACTH levels in the group are 1595 ± 365 and 1152 ± 183 (pg / ml), and the corticosterone levels are 845 ± 48 And 786 ± 24 (ng / ml). The effect of EF exposure on piasma parameters. Figures 14A and 14B show that EF exposure is constrained in normal (a) and Indian nested (B) rats. Effects of Changes in Plasma Glucose Levels Induced. Those levels were detected after a period of 60 minutes (n = 6). The number of samples was 6 in all groups. Data are expressed as a median, 25th to 20th percentile, 75th percentile, minimum, and maximum. *: The corpse remains from the unprocessed group. In ovariectomized rats, plasma glucose levels were constrained to increase (P < 0.05: Stuart's t test), and ef or diazepine tended to suppress these increases (Figure 14B). However, the tendency to suppress the plasma glucose 38 200423986 level in this EF cohort was not found in normal rats that did not undergo an ovariectomy (Figure 14A). Figures 15A and 15B show the effect of EF exposure on the plasma lactate level induced by restraint in normal (A) and ovariectomized (B) rats. The 5 levels were measured after a 60-minute period (n = 6). Data are expressed as a median, 25th percentile, 75thI quantile, minimum, and maximum. *: Corpse < 0.05 from untreated group.卞: The corpse < 0.05 was from the simulated group. In ovariectomized rats, plasma lactate levels in the restricted-only cohort did not show significant differences compared to the untreated cohort (Figure 15B). Plasma lactate levels in the EF-exposed and administered diazepine groups were significantly lower than those in the restricted-only group (/><0.05: Stuart's {test) (Figure 15B) ). In normal rats, the plasma lactate levels (mean ± S.E.) were 28. 6 ± 3. 6 and 38.1 ± 37 (mg / dl) with and without EF (Figure 15A). As shown by the results of a statistical analysis, the lactate levels of animals exposed to 15 EF were significantly lower than those in this constraint-only group (P < 0.05: Stuart's t-test). Figure 16 shows the effect of EF exposure on the plasma pyruvate level induced by restraint in ovariectomized rats. The level was measured after a 60 minute period (n = 6). Data are expressed as a median, 25th-20th percentile, 75th percentile, minimum, and maximum. *: From a group without processing. In the ovariectomized large salamander, the plasma pyruvate level of the only-constrained group was not significantly different from those in the untreated group, but tended to decrease due to the restriction. Individual systems in the cohort exposed to £ 17 or administered to diazepine were significantly lower than those in cohort 39 200423986 of simulated EF exposure (P < 0.05: Stuart's t test) ( (Figure 16). Figure 17 shows the effect of EF exposure on confinement-induced white blood cell (WBC) counts in ovariectomized rats. The level is measured after a period of 60 minutes and 4 (n = 6). The data are expressed as a median of 5 digits, 25 & percentiles, 75 percentiles, minimum and maximum. *: P < 0 · 05 is from a group without processing. In general, the observed constraint-dependent changes are related to the number of white blood cells (WBCs). The WBC counts in the untreated, only constrained, EF-exposed, and diazepine-administered groups showed 78, 99, 96, and 85 (x 102 cells / μΐ), (Figure 17). As shown by the results of a statistical analysis, in the ovariectomized rats, the WBC level in the restrained animals was significantly higher than in the untreated group (p < 005: Stuart's t test ). The WBC level tended to be higher in the EF exposed or administered diazepine groups than in the untreated group and lower than in the restricted group only. 15 1 Example 7-Electroencephalogram Studies Six rats were exposed to an electric field estimated at 17,500 V / m for a period of 15 minutes for a total of 7 days. The device used to expose the animals was a Healthtron exposure cage (as previously described). Six rats were used as a control group (simulated exposure). The following parameters (endpoints) were observed: 20 brainwave abnormalities detection; each EEG stage group (wake, rest, siwave wave Hght sleep), Slow wave deep sleep and percentage of fast wave sleep; and EEG power spectrum of frontal cortex δ (1-3.875 Hz) ^ Q (4-15.875 Hz) > a (8-12 Hz)-β 40 200423986 5 10 1 (12 · 125-15 · 875 Hz), and a percentage of / 5 2 (16 · 25 Hz). When # ~ exposed to 7,000 V (17,500 V / m) for 15 minutes, an increase in the slow-wave light sleep sleep stage was observed over a period of 1-2 hours, which was significant over the first day. On the seventh day, a significant reduction in rest periods and awake periods was observed at 4 ° after exposure. A significant decrease in awake periods and a significant increase in island sleep periods were observed over a period of time varying from 0.5 ^ after exposure. A significant decrease in the awake period and a significant increase in the slow-wave deep sleep sleep period were observed over a period of time that varied from time to time after exposure. In addition, a significant increase in the slow wave light sleep period was observed over a period of time ranging from 2-4 hours after exposure. No spontaneous EEG wave type or behavioral abnormalities were observed. There was no evidence in this study that repeated exposure to an electric field showed any neurological relationship to frequency analysis of the frontal cortex in rats. 15 D. Additional disorders or conditions

To treat electrolyte imbalance, the average induced current density generated across the cell membrane is preferably about 0.4 mA / m2 to about 6.0 mA / m2, and more preferably about 0.4 mA / m2 to about 5 6 mA / m2, and still more preferably about 0.43 mA / m2 to about 5.55 mA / m2. 20 For the treatment of arthritis, the average induced current density generated across the cell membrane is preferably about 0.02 mA / m2 to about 0.4 mA / m2, more preferably about 0.025 mA / m2 to about 0.35 mA / m2, Most preferably about 0 026 mA / m2 to about 032 mA / m2 ° For the treatment of overweight, the average induced current generated across the cell membrane 41 200423986 density is preferably about 0.002 mA / m2 to about 1.5 mA / m2, more preferably about 0.02 mA / m2 to about 1.2 mA / m2, most preferably about 0.024 mA / m2 to about 1.12 mA / m2 ° The present invention can also be applied to prevent or treat musculoskeletal and connective tissue Obstacle 5 obstacles. These disorders include, for example, osteoporosis (including senile, secondary, and juvenile idiopathic), bone-thinning disorders, celiac disease, tropical diarrhea ( tropical sprue, bursitis, scleroderma, CREST syndrome, Charcot's 10 joints, proper repair of fractured bones, and ligaments and cartilage (Cartilage) Proper repair of the tear. The present invention can also be applied to rheumatoid arthritis, immunosuppression disorders, neuralgia, insomnia, headache, facial paralysis, and mental officer 15 Neurosis, arthritis, joint pain, allergic rhinitis, stress, chronic pancreatitis, DiGeorge abnormality, endometriosis, urinary Urinary tract obstructions, pseudodogout, thyroid disorders, parathyroid disorders, hypoptosis, 20 hypopopituitarism, gallstones, peptic ulcers, salivary disorders salivary gland disorders, appetite disorders, heart palpitations, vomiting, thirst, excessive urine production, vertigo, benign paroxysmal positional vertigo, Esophagus 42 200423986 achalasia and other neurological disorders, acute kidney Dried (acute kidney failure), chronic renal failure (chronic kidney failure), rhesus diffuse esophageal spasm (diffuse esophageal spasms), and transient cerebral ischemia (transient ischemic attacks, TIAs). The present invention is also applicable to the treatment of conditions or disorders including osmolarity, additional renal disorders that it maintains, and an osmolar imbalance. E.EF Therapy Device The EF device is designed to generate an electric field in which an individual is placed. As illustrated in Figure 18, this electric field can surround the entire individual. Alternatively, the electric field may surround only a particular part or organ of the individual. Fig. 19 is a schematic diagram of a high voltage generation device (1) showing a specific example of the present invention. That is, the potential therapy device (1) includes a potential therapy device (2), a high voltage generating device (3), and a commercial power source (4). The potential therapy device (2) includes a chair (7) with armrests (6) for a body (5) 15 to sit on; a head electrode (8), which is attached to the upper end of a cymbal chair and An opposite electrode provided above the top of the individual's head (5); and a second electrode (9), such as an ottoman electrode, which is a main electrode, and the individual (5) places his / her legs on the first The top surface of the two electrodes. It is noted that the head electrode (8) is an opposite electrode of the second electrode (9) (its 20 is a main electrode), and may be another ceiling, wall, floor, furniture or other room Internal objects or parts. The high voltage generating device (3) generates a voltage to apply a-voltage to the head electrode (8) and the second electrode (9). The high voltage generating device ⑶ is usually installed below the chair ⑺, between the legs and on the floor, or near the chair ⑺. 43 A distance (d) between the f-th pole of the -M part and the patient's head can be changed. An insulation material surrounds the chanter electrode (8) and the second electrode (9). The second electrode (9) is connected to a high voltage output terminal (10) of the high voltage generating device (3) through a wire (11). It also has a high voltage output terminal (10) to apply a voltage to the head electrode (8) and the second electrode (9). In addition, the chair (7) and the second electrode (9) include insulators (12), (12) at a position in contact with the floor. The distance ⑷ between the surface of the human body and the first electrode (8a) can be easily changed by placing cushions of different thicknesses on the bed base ⑼. 10 A potential treatment device (2C) still provided with another structure has a type shown in Figure 20A [perspective view] and Figure 20B [side view], which exemplifies the individual (5) and each subject Chair shape in the positional relationship between the electrodes drawn in black]. The chair (7a) is provided with a front open cover body (34) covering the individual (5). The covering body (M) is provided with a fifteenth electrode (8c), which is used as an opposite electrode for accommodating the head of the individual (5); a second electrode (9c), which is used as a main electrode. Oman electrode; and another first electrode (80c) provided at a position from the shoulder to the waist when in a sitting position, as an opposing electrode provided at the waist of a higher part of the body. The other first electrode (80c) has a plurality of side electrodes (80c,) to cover the body of the individual (5) from the 20 sides. Preferably, the first electrode (8c) is arranged along the head of the human body, and the other first electrode (80c) is arranged in several layers from the shoulders to the waist along the longitudinal direction. These first electrodes (8c), another first electrode (80c), the side electrodes (80cf), and the second electrodes (9c) are arranged in an insulating material (35). A detachable cushion member (2004) made of an insulator is attached to the cover (34). Therefore, the attachment of a pad component can be changed with different thicknesses. I: The distance between the surface of the human body and the first electrodes (8c), (80c), (80c,). As also mentioned above, in this kind of potential therapy device (2c), the responsive 5 current control member can be applied to the first electrodes (8c), (80c) which are applied as a counter electrode. , (80c '), and the second electrode (a voltage is applied on the ninth phantom, and is between the first electrodes (8c), (80c), (80c,) and the torso surface of the human body The distance (d) between them is variable or is applied to the first electrodes (8c), (80c), (80c,) and the second electrode (9c) by controlling 10 Applied voltage, and further, controlling the electric field on the surface of the human body by changing the distance ⑷ between the first electrodes (8c), (80c), (80c ') and the surface of the human body A very small amount of induced current flows in various regions of a human body's torso. A potential therapy device (2A) provided with another configuration is shown in Figures 15-21A [perspective view] and 21B [side view] The potential therapy device (2A) has a bed type. A box (32) for accommodating the individual (5) is provided on a bed base (31). Each electrode is provided here Sub (32). In short, it is provided with a first electrode (8a) as a counter electrode and a second electrode (9a) placed on a leg of the human body as a main electrode. The first electrode 20 (8a) is placed on the head, shoulders, abdomen, legs, buttocks or other areas of a human body. Preferably, the first electrode (8a) has a head, shoulders, abdomen, and The shape, width, and area of the buttocks. The blank areas in these illustrations show where no electrodes are placed. The electrodes are placed in an insulator (33). A cricket 45 made of an insulator (not shown) 200423986 was placed on each electrode of the bed base (31). Mats of different thicknesses were prepared. In the above-mentioned figure 19, the head electrode (8) above the head and the individual (5 The distance (d) between the torso surfaces of the human body is set to about 5 to 1 to 25 cm. In Figure 20A, the first electrodes (8c), (80c), (80cf) ) And the torso surface of the human body of the individual (5) is set at about 1 to 25 cm, preferably 4 to 25 cm, and in Figure 21A, the distance (d) between the first electrodes (8a), (8b) and the torso surface of the human body of the individual (5) is set to about 1 to 25 cm, preferably about 3 to 25 cm. 10 As described in Figure 22 below as an electronic configuration block diagram, the high voltage generating device (3) has a voltage for one of the commercial power sources. 100V AC is increased to, for example, 15,000V booster transformer (booster tmnsfomier) (t), and current limitation resistors (R), (R) to control the current flowing to each electrode f. The high voltage 15 generating device (3) has a configuration in which an intermediate point (s) of a booster coil (T) is grounded and the ground voltage is set to a boosted voltage. half. As shown by the illustrated temporary line, a point (sf) can be grounded. Here, as shown in the block diagram in Fig. 22, a high voltage (its intermediate point (s) on the high voltage side is grounded by the boost transformer 20 (bottom)) from a 100V AC power supply via Obtained by a voltage controller (13) of the high voltage generating device (3), and further, each high voltage passes through the current limiting resistors (R), (Rf) for protecting human body And connected to these head electrodes (8), (8c) or the like (see below), and these second electrodes (9), (9c) or the like 46 200423986 (see below ). Furthermore, the potential therapy device is provided with an induced current control means. This induced current control member can cause a very small amount of induced current to flow to various regions constituting the individual (5), the human body's torso, and is applied to the head electrode ⑻ and the second electrode (9) by changing ^ 5 pressure, and-the distance between the head electrode ⑻ and the dry surface of the human body face ⑷ 'or by applying control to the head electrode ⑻ and the second electrode (9) The external pressure is controlled by changing the distance between the head electrode ⑻ and the stem surface of the human body to control the electric field lying on the body. The distance between the surface of the human body and the first electrode (8a) can be easily changed by placing, for example, a pad of the same thickness on the bed base (31). By increasing the induced current, even in a state where a high voltage is applied to the potential therapy device (1), a higher therapeutic effect can be obtained, even if it lasts for the same period of time as in the conventional method. In addition, the 15 treatments can be completed in a shorter time than before. Furthermore, in order to obtain the same therapeutic effect, an induced current of the same value as in the prior art can be obtained at a lower voltage and at the same treatment time as before. The potential therapy device (1) of the present invention is designed to be as free as possible from high-output electronic noise, high-level radio frequency noise, and strong magnetic fields. In order to reduce the influence of electromagnetic field interference on the potential therapy device (1), it is better to use a driving mechanical switch, a relay and an electric motor, or an electrical timer or other electrical components than electronic components, semiconductors, Power components (such as thyristor, triac) 47 200423986 electronic timer or EMI sensing microcomputer for attack and manufacture. However, 'such as electronic functional components, electronic serial bus switching regulators are effective as optical emitter diode power supplies, and 5 this optical emitting diode is used as An optical source that informs the individual or operator of the active or inactive state of the potential therapy device of the present invention. As discussed above, a simulated human body (h) can be used to measure IEF and induced current. , As shown in Figures 23A, 23B and 23C. 10 This simulated human body (h) is made of PVC and its surface is coated with a mixed solution of silver and vaporized silver. This results in resistance (1K Ω or less) equivalent to that of a real human body. The simulated human body (h) is well known as a nursing simulator (nursing simulat〇r), and its size is similar to an average human body size, for example, its line is 174 (^ 1 high. The I5 temple size is further It is described in Table 1. 48 200423986 Table 1: Section of Area that simulates the current density in the human body. Circle (mm) Cross Sectional Area (m2) Eye 550 0.02407 Nose 475 0.01795 Neck 328 0.00856 Chest 770 0.04718 Upper abdomen 710 0.04012 Arm 242 0.00466 Wrist 170 0.00230 Torso 660 0.03466 Thigh 450 0.01611 Knee 309 0.00760 Ankle 205 0.00334 Electric field on the surface of the body by attaching a disc-shaped electric field measurement sensor (e) to the simulated human body (h) is measured in a measurement area. These measurements occur at 115V / 60HZ and 120V / 60HZ. 5 A method for measuring an induced current and a device for this purpose are shown in Figure 24 In the inductive current measuring device (20), as shown in Figures 23A and 23B, the simulated human body (h) It is placed on the chair (7) in a normal sitting position. The head electrode (8) (which is the opposite electrode) above the head is adjusted and installed on a head from the simulated human body (h) 10 square llcm. These measurements are performed by measuring various portions (such as, for example, the k-kf line portion illustrated in Figure 24), transferring the induced current waveform through optical tmnsfer, and This waveform was achieved by observing this waveform on the ground side of the current measurement device (20). Here, the applied voltage was 15,000 V. In this measurement method, measurements were made in 15 regions of the simulated human body (h). The induced current is obtained by using two leads to generate a short circuit that flows across the simulated human body's puppet part 49 200423986 (s.hort-ci: rcuit) (22) [not shown] to obtain the induced current. Measured The resulting induced current is converted into a voltage signal via an I / V converter (23) (Figure 24). This voltage signal is then converted into an optical signal via an optical analog data link on the transmission side. 5 These optical letters The signal is transferred via an optical fiber cable (25) to an optical analog data link (26) on the receiving side and converted into a voltage signal. This voltage signal is then processed by a frequency analyzer (27), and frequency analysis is performed by a waveform observation and analysis recorder. A buffer and a 10 adder are placed between the I / V converter (23) and the optical analog data link (24) on the transmission side [not shown]. Therefore, the measured electric field values and induced currents at 115 v / 60] ^ and 120 v / 60 Hz at the locations of various regions of the simulated human body (h) are shown in Table 2. If the electric field value is different from Table 2 'of this table, then it is known that the value of the induced current passing there is also different. Therefore, it is assumed that the effective induced currents for each area where a real human body lies dry can obviously be obtained by changing the electric field of each area involved. 50 200423986 Table 2: Relationship between the electric field value and the induced current value @ TT5V / 50Hz @ 120 V / 60Hz Electric field value (kV / m) Induced current U a) Electric field value (kV / m) Induced current (UA) Face 182 0.72 190 0.90 Front 81 0.32 84 0.40 ^ 1_Second-Back of M section 113 0.44 118 0.55 Face 16 0.06 " 16 0.08 Shoulder 37 0.15 38 0.18 19 0.08 20 0.10 29 0.1Ϊ 30 0.14 33 ^ 0.14 34 0.17 _ 52 0.20 " " 54 0.25 surface 21 0.08 22 0.10 ____ 42 0.17 43 0.21 11 0.05 12 0.06 21 0.08 22 0.10 end 3.4 0.01 3.5 0.02 348 1.37 363 1.72 The body surface electric field E can be displayed by using the following equation, The induced current value of each region obtained by the method of measuring the induced current of each region in FIG. 24 is obtained. That is, E = I / soooS. Here, S series 5 is all surfaces of the electric field measurement sensor, εο is an induction rate in a vacuum, I is an induced current, ω is 2jtf and f is frequency. When the induced current in each area is obtained by the above method, an induced current density j in each area can be obtained using the following formula. That is, A = 2πΓ, B = jcr2, B = Α2 / 4π, and J = I / B, where A is a circle of 10, B is a circular area, r is a radius, and I is a measurement The measured current, and j is an induced current density. When the potential therapy is performed by controlling the voltage of the head electrode (8) and the external voltage applied to the second electrode ⑼, the temple-induced current control member mentioned above can cause—a very small amount of Induced currents flow to various areas of the trunk of a human body. 51 200423986 Table 3 shows the relationship between: (i) the induced current in the nose, neck and torso (μ A); (2) the induced current density in the nose, neck and torso (mA / m2) ; And the applied voltage (KV) at 120V / 60HZ. At the same applied voltage, this current density tends to be highest in the neck, second highest in the trunk 5 and lowest in the nose. It is noted that the induced current density in Table 3 is less than 10 mA / m2, and the current density of 10 mA / m2 or less has been approved by the International Commi ssion on Non Ionizing Radiation Protection ) Is established as safe. Table 3: Applied voltage and induced current Applied voltage [kV] Induced current value (// A) Induced current density (mA / mz) Head (nose) Neck torso Head (nose) Neck torso 0 0 0 0 0.0 0.0 0.0 5 10 11 30 0.6 1.3 0.9 10 20 23 61 1.1 2.6 1.7 15 30 34 91 1.7 3.9 2.6 20 40 45 121 2.2 5.2 〇25 50 57 152 2.8 6.6 4Λ 30 60 68 182 3.3 7.9 Γ 2 10 Figure 25 also shows the relationship between the applied voltage (κν) and the induced current (// A) in the nose, neck, and trunk. As apparent from Fig. 25, 'the applied voltage and the induced current are proportional to each other. Table 4 shows the change in induced current and induced current density in a human neck as a function of the distance (d) between the head electrode (8) and the top of the head. 52 200423986 Table 4: Change in induced current as a function of distance from the electrode. Distance between the first electrode and the top of the head. Induced current value. Induced current density distance (cm) (/ z A) (mA / mz) 4.3 50 5.8 5.4 46 5.4 6.3 43 5.0 6.9 40 4.7 8.3 39 4.5 9 38 4.4 9.9 35 4.1 11 34 3.9 12 34 3.9 13 33 3.8 14 31 3.7 15 30 3.5 16.1 30 3.5 17.2 30 3.5 Table 4 indicates that in a 15cm or more Distance, the induced current is stable at 30 μΑ. Therefore, in order to change the induced current by changing the distance, the distance should be 15 cm or less. Figure 26 also shows the change in induced current due to the distance (d) 5. In an experiment involving approximately 300 human cases of low back pain, we determined that EF is effective in treating low back pain. We also determined the optimal dosage and parameters as follows. In short, the optimal dose is obtained by controlling the generation of the induced current value and the transit time of the induced current through the area that constitutes a person's body lying dry. In addition, it is obtained by controlling the generation of a sum of applied voltages other than the first electrode voltage and the second electrode voltage, and the application time thereof. For low back pain, the therapeutic efficacy of EF is optimized by administering it at a voltage of about 10 KV to about 30 KV (preferably about 15 KV) for about 30 minutes. In other words, at about 300 KV / min to about 900 KV / min, preferably 15 about 450 KV / min. Here, Table 5 shows the values of the induced currents measured at 115 V / 50 Hz in the 53 domain W points of each region constituting the simulated human body (h) torso, and considering the simulated human body in Table 1 Under the size, the induced current density obtained from the difference of this induced current value. From Table 5, the measured values of the induced current (μΑ) and the calculated values of the induced current density (mA / m2) in the various regions constituting the torso of the human body are as follows: eyes: 18 / 0.8; nose: 24 / 1.3; Worker 4.27 / 3.1; chest: 44 / 0.9; pit 〇f the stomach: 8.6 / 1. 6; and torso: 91 / 2.8. Range, induced current value, and induced current density part of the induced current @ 115V / 50Hz 〇A) Induced current density @ 115V / 50Hz (mA / m2) Eye 18 0.8 Nose 24 1.3 Neck 27 3.1 Chest 44 0.9 Stomach depression 65 1.6 Arm Arm 8.6 1.8 Wrist 3.1 1.3 Torso 73 2.1 Thigh 46 2.8 Knee 52 6.8 Ankle Ankle 58 17 In addition, based on the aforementioned induced current and induced current density, the induced current and induction at 1210 V / 60 Hz The current density is calculated according to the following equations and 2. Formula 1: Induced current: I (60Hz) = I (50Hz) x 60 / 50x 120/115 15 Formula 2: Induced current density: J (60Hz) = J (50Hz) x 60 / 50X 120/115 54 200423986 Table 6 shows the calculation results of the induced current and induced current density in various regions of the human body trunk at 120 v / 60 Hz. From Table 6, the measured values of the induced current (μΑ) and the calculated value of the reduced deer current density (mA / m2) in each area constituting the trunk of the four people are as follows: eyes: 23 / 〇 · 9; nose · 5 30 / 1.7; Neck: 34 / 3.9; Chest: 55A.2; Stomach depression: 11/2 3; and Torso: 114 / 3.6. Table 6: Area, induced current value, and induced current density part of the induced current @ 120V / 60Hz (# A) Induced current density @ 120V / 60Hz (mA / m2) Eye 23 0.9-Nose 30 1.7 Neck 34 3.9- Chest — 55 1.2 Small stomach depression 81 2.0 11 2.3 3.9 1.7 Lying dry 91 2.6 Thigh 57 3.6 Knee 64 " 83 ~~ — Foot sorrow 72 22 When the distance between the electrode and the human body area is fixed, the above & and the applied voltage flowing through various areas of the body of a human body is proportional to 10 induced currents. Therefore, when a human body is treated with a chair, the optimal dose can be obtained by controlling the generation and application time of the applied voltage, because if the distance between the electrode and the human body is The method of the best common divisor is determined, then the electric field strength of each area of a human body is almost determined by the externally applied voltage. A trained individual will understand that the amount of applied voltage and current density can be controlled using an appropriate electric field device, such as a Healthtrón 55 200423986 HES-30 ™ device (Hakuju Co.). For example, the induced current generated in a biological sample can be increased by increasing the potential of the electrode by being applied through EF. Other suitable devices are known to trained individuals and include, but are not limited to, 00298 device 5 (Hakuju Co ·), HEF-K 9000 device (Hakuju Co ·), and HES-15A device (Hakuju Co.), HES-30 device (Hakuju Co.), AC / DC generator (Sankyo, Inc.), and function generator SG4101 (Iwatsu, Inc.). The characteristics of some exemplary devices are presented in Table 7 along with specifications for those devices. 10 Additional electric field devices that can be used with the method of the present invention include the electric field generating device disclosed in U.S. Patent No. 4,094,322, the entire contents of which are incorporated herein by reference. The treatment device is capable of directly transmitting an electric field to a desired portion of a patient lying on the device. Other electric field devices are disclosed in U.S. Patent Nos. 4,033,356, 4,292,9880, 15 4,80,2,470 'and British Patent GB 2 274 593, the entire contents of each of which are incorporated herein by reference. Table 7 provides special specifications for the EF device that has been selected for use with the method of the present invention. 56 weight high voltage group I 40 kg body 41kg 130 kg 240 kg insulation pad (N treatment chair with power off switch box 15 kg 00 chair [15.8 kg -M bJQ (N control switch box ω ω -4 < automatic timing Device time 30 minutes +/- 10% 30 minutes with 1, 2, 4, 6, and 8 hours unlimited output voltage Charging Footrest > ^ N Charging Footstool o rn ό > 0-15,000 V 0-30,000 V > > 88 ss ϋ u < Q AC: 0-3,500 V; DC: 0-3,500 V Upper Electrode .u > gtc 82 n 「Upper electrode > o to en ό Power 18VA +/- 15% 10 W 100 VA 200 VA 25 W 25 W Rated power supply frequency 60 Hz 50 or 60 Hz 50 or 60 Hz i 50 or 60 Hz 50 or 60 Hz 50 or 60 Hz rated power supply Voltage 115 V AC 100 V AC 100 V AC 100 V AC 100 V AC 100 V AC Device Type I 00298 HEF-K 9000 HES-15A HES-30 AC / DC Generator Function Generator: SG4101 200423986 In homogeneous but external appearance The current-density distribution induced by a 60-Hz electric field in an irregular human model is A two-stage finite difference procedure (twc) -stage finite-difference procedure (Hart, FX ·, 11: 213-228 (1990)) was calculated. For 5 ungrounded exposed to a 10 kV / m electric field In the case of the human model, the induced current density in the plane passing through the height of the lower back of the torso is 1 · 14 mA / m2 (Figure 27). The current density in other locations is in the range of 0.8-3.5 mA / m2. Correct The value depends on the coupling capacity between the model and the ground (capacitive coupling), but a reasonable coupling condition range of 10 results in a change in the calculated current density of less than a factor of 2. Similar results are obtained by Others have found (Gandhi, OP & Chen, JY ·, Bioelectromagnetics Suppl. 1: 43-60 (1992); King, RWP ·, / 555-7> fl muscle Eng 45: 520-530 (1998)) . The finite-difference time-domain method is used by 15 to calculate the induced current in the basic anatomy model of the human body (Furse, CM & Gandhi, OP ·, Bioelectromagnetics 19: 293-299 ( 1998)). This calculation is performed in a supercomputer to allow a much larger amount of parsing than previously possible. The results obtained from the current density induced in the special tissue of the model are shown in Table 8. Comparable results have been used by others including fat-muscle (Chuang, H.-R · & Chen, KK-M ·, / 55 May Tram £ ^ off · 36: 628-634 (1989)) and bone -A mixed tissue model of the brain (Hart, FX · & Marino, Α · Α "Mel BwZ · 5ng · Camp · 24: 105-108 (1986)) was discovered. 58 200423986 Table 8 · Exposure to 60Hz Current density induced in a human individual special tissue at 10 kV / m electric field Tissue induced current density (mA / m2) Small intestine 1.3 Spleen 1.4 Pancreas 1.5 Liver 1.4 Kidney 2.8 Lung 0.6 Bladder 1.9 Heart 2.2 Stomach 1.2 Testicle 0.7 Prostate 1.0 Eye humor 5.6 Cerebrospinal fluid 4.8 Pineal gland 1.4 Pituitary gland 3.5 Brain 1.9

Example 8-Exposure to Electric Field (EF): It has some clinical symptoms in human patients

的 ## Palliative Effect. 5 Electric field exposure device, Healthtron (Model HES 30, Hakuju

Institute for Health Sciences Co., Ltd., Tokyo, Japan) is used. Healthtron includes a step-up transformer (a device used to control the voltage in a circuit), a seat, and electrodes. It applies a high voltage to one of the two opposite electrodes to produce a predetermined potential difference 10 and forms an EF in the space between the two electrodes. The user sits comfortably and is allowed to read a book or sleep during the exposed time. In order to avoid accidental electric shock caused by the formation of electrical current, 59 200423986 During treatment, these individuals are not allowed to make any form of physical contact with the floor and anyone (operator and other people exposed to electricity). Insulator-covered electrodes are placed on the floor that allows feet to rest on, and on the heads of individual patients. A starting power of 30,000 volts (ELF at 50 or 60 Hz) is applied to the electrode placed on the foot, producing an EF between the foot and the head electrode. It took 30 minutes for the track record to reach the mother's stage during the road crash and the frequency of exposure changed from once a day to once a week.

The effectiveness of Healthtron is based on the direct supervision of Dr. Yuichi Ishikawa at Toranomon Clinic 10 Minato-ku (Tokyo, Japan) in Minato-ku, Tokyo from August 1, 1994 to June 30, 1997. The results obtained from the questionnaires submitted were evaluated. A total of 1,253 patients (489 men, 764 women) performed the device, of which 505 (208 men, 297 women) visited the clinic and used the Healthtron device and implemented the device at least twice. Others can use the 15 device more than twice. To reduce the subjectivity of the participants in the questionnaire, the assessment of Healthtron's moderating effect was limited to these 505 patients. Each Healthtron user was taken care of by a doctor and interviewed for the moderating effects of the device during previous visits. The interview included inquiries regarding major physical discomfort (= symptoms), past medical history and treatment, frequency of use of Healthtron20, and perception after use (including its moderating effect and the user's personal healthtron holding) . The severity of symptoms during the first hospital visit was rated on a scale of 3, and the severity after Healthtron therapy was divided into 5 scales: very good (5), good (4), no change ( 3), deterioration (2), and highly deterioration (1). Very good and good are distinguished as "Easy 60 200423986 'regardless of the frequency / interval of exposure, and the time of ease (in days) is also recorded. As a result, the age distribution of patients was between 20 and 90 years old Among them, 85.3% were made up of> 40 years and 5 age groups (Table 9). There were 208 (41%) men and 297 (59%) women. 55 different symptoms were identified, and each of them The proportion of patients reported to be relieved by Heahhtrn therapy is summarized in Table 9 "Response. Symptoms identified by at least one patient include: cold feeling in the limbs, fatigue, headache, High blood pressure, insomnia, 10 knots pain, lower back pain, pain in the limbs, pruritus cutaneous, paralysis of the limbs, shoulder / neck pain and stiffness. The alleviating effect of Healthtron therapy is significant in the following systems: no accompanying fever Headaches, organotherapy (such as subarachnoid or cerebral hemorrhage)

Hemorrhage)), or inflammation (91.7%), joint pain (60.7%), lower back pain 15 (57.3%), shoulder / neck pain and stiffness (56.0-57.8%), and reduction of fatigue (55.0%) . Interestingly, the mitigating effect on pain-related symptoms affecting locomotodal organs (head, joints, shoulders, neck, limbs, and abdomen) was recorded as 175 (58.5%) of 299 cases. These pain-related symptoms do not result from traumas. After the first 20 treatments, 4 of the 10 patients with itchy skin claimed to have been relieved 'and the clinical manifestations of one patient were exacerbated. 61 Table 9: Age range and gender distribution of Healthtron users Number of users in age range Male: Female to 20 2 2: 0 21 to 30 38 15:23 31 to 40 34 10:24 41 to 50 81 29:52 51 ～ 60 147 59:88 61 ～ 70 143 69:74 71 ～ 80 50 20:30 81 ～ 90 10 4: 6 Total 505 208 (41%): 297 (59%) Table 10 shows that among 505 patients, 55 A confirmed clinical symptom of 200423986 is the Palliation Rate. Table 10-Remission rates of 55 clinical symptoms in 505 patients. Number of patients with symptoms (%) Abdominal fullness 1 0 (0) Abdominal pain 2 1 (50) Allergic constitution 7 3 (42.9) Alopecia 3 3 ( 100) arrhythmia 2 1 (50) back pain 5 3 (60) blurred vision 5 2 (40) chest pain 1 1 (0) cold feeling in the limbs in the extremities) 14 6 (42.9) constipation 5 3 (60) cough 5 3 (60) deafness 2 1 (50) diarrhea 3 3 (100) dizziness ) 5 3 (60) ear ringing 7 1 (14.3) If (enervation) 4 3 (75) | exanthema 4 1 (25) 62 200423986 eyestrain 5 1 (20) face Facial edema 1 1 (100) Facial numbness 2 0 (0) Facial paralysis 1 1 (100) Facial stiffness 1 0 (0) Fatigue ) 20 11 (55) generalized muscle stiffness 1 〇 (〇) gingival pain 1 0 (0) glycosuria 7 4 (57.1) Headache 12 11 (91.7) Heavy feeling in the body 4 2 (50) Heavy feeling in the head 1 〇 (〇) Heavy feeling in the legs ) 1 1 (100) Heavy stomach feeling 1 〇 (〇) Hypertension 10 4 (40) Insomnia 17 8 (47.1) jaundice 1 1 (100 ) Joint pain 45 30 (66.7) loss of appetite 1 0 (0) loss of grip 1 0 (0) lower back pain 89 51 (57.3) menstruation Menstrual irregularity 1 〇 (〇) pain in the extremities 31 10 (32.3) Palpitation 1 1 (100) paralysis in the extremities 3 〇 (〇) plantar edema ( plantar edema) 4 2 (50) Pollakiuria 1 1 (100) pruritus cutaneous 10 4 (40) 63 200423986 stiffness of the arms 1 1 (100) limbs sensation of numbness in the extremities) 29 11 (38.0) separation of the. calx epidermis 1 1 (100) Shoulder or neck pain 25 14 (56) Shoulder or neck stiffness 90 52 (57.8) Sore throat 2 1 (50) Stomachache 5 4 (80) swelling of joints 2 2 (100) trembling of the extremities 1 1 (100) urinary incontinence 1 〇 (〇) Total 505 268 (53.1) Figure 28 Shows the average time to remission of each symptom in 505 patients regardless of the frequency / interval of Healthtron therapy. Considering the small sample size of many of the identified symptoms, in an inherent limitation of this study, the researcher relied solely on data from the questionnaire, and we believe that the continuity of the moderating effect of treatment is limited to those Symptoms confirmed by at least 10 patients showing a> 50% remission rate can be properly described. The relaxation of fatigue lasted for about 50 days; joints, lower back and shoulder / neck stiffness were alleviated and said to be less than 100 days. The longer average relaxation time noted in many other symptoms may be a sample-size response rather than the true utility of the treatment. F. Method of optimizing electrotherapy parameters The selection and control of the parameter range of the present invention while avoiding unnecessary side effects that may be caused by the use of EF 64 200423986 makes it possible to use EF as a treatment tool. Therefore, the present invention provides parameters and the range in which they are used to enable a trained individual to use EF as a treatment tool to achieve a specific biological result and avoid unnecessary side effects. 5 A preferred method for determining the optimal parameters of EF therapy includes the following steps: (i) confirming a desired biological response to be induced in a living organism; (ii) selecting or measuring a The average induced current density on the cell membrane of an organism or a tissue sample or cell derived from the organism is 10 degrees; (ϋ 〇 Select or measure an external electric field, which is away from the organism, sample or A selected distance is measured or measured at a specific distance of the culture; (iv) a continuous time period is selected or measured to produce the selected current density at the cell membrane; (v) Applying the selected or measured a: electric field to the organism, sample, or culture, so that the selected or measured induced current density is generated on the membranes of the cells for a continuous period of time of the selected or measured; (Vi) determining the extent to which the desired biological response occurs; (Vii) selectively repeating any of steps (ii) to (Vi); and (viii) confirming that the desired biological response is best induced The response should be selected or measured The induced current density, the selected or measured external electric field, or the value selected during the continuous time of 20 measurements. Preferably, the method further includes generating a dose-response curve for uranium 'in step (viii) (Dose-response curve) as a function of any of the selected or measured induced current density, the selected or measured external electric field, or the selected or measured continuous time period. Still more preferred The method further comprises, before step (viii) 65 200423986, selecting or measuring the following: the number of times step (V) is repeated, the time interval between steps (V) repetition, and the selected or measured The induced current density is generated throughout the entire period of the films. More preferred specific examples include one or more of the following characteristics: The selected 5 or measured induced current density is about 0.001 mA / m2 to About 15 mA / m2; the induced current density measures the induced current flowing through a specific area of a living organism or a part thereof, converts the detected current into a voltage signal, and converts the voltage 彳 § 5 tiger to make An optical signal, and then the optical signal is converted into a voltage #, and the waveform and frequency are analyzed and selected or measured. 10 and / or the external electric field (E) is expressed as E = I The terms in / socoS are selected or measured, where s is the section of an electric field measurement sensor, εο is an induction rate in a vacuum, and I is A current, and £ 〇 [0S is 2jxf £ should be 0] is 2πί, see the explanation 1 of the original English p-3, and f is the frequency. 15 One is used to determine the best external current therapy The preferred method of parameters includes the following steps: (i) confirming a desired biological response to be induced in a living organism; (ii) selecting or measuring a bit in the organism or a derivative of the organism The average induced current density on the cell membrane of a tissue sample or culture cell is 20 degrees, where the average applied current density is about 10 mA / m2 to about 2,000 mA / m2; (iii) selecting or measuring one will produce The selected or measured current of the applied current density; (iv) selected or measured During continuous time, 俾 to produce the selected or measured applied current density; (v) apply the selected or measured current to produce the selected or measured applied current density for 25 seconds to be selected or measured (Vi) determine the degree of privacy that the desired biological reaction 66 200423986 should occur, (vii) repeat any one of steps (i) to (vi), and generate a dose-response curve as A function of the selected or measured current, the selected or measured applied current density, or the continuous time period of the selected or measured; and (viii) confirming that the desired biological response is best induced The selected or measured current, the selected or measured applied current density, or the value of the selected or measured continuous time period. Preferably, the method further comprises, before step (viii), selecting or measuring the following: the number of times step (V) is repeated, the time interval between step repetitions, and the applied current density is generated throughout the The full period of the isomery. O The inventors have determined the parameters that best treat a particular disorder. All in all, the EF voltage (exogenous) can be applied in a range between about 50 v to about 30 kv. The induced current density can be generated in a range between about 0.001 to about 15 mA / m2. Preferably, the EF induced current density is generated in a range of about 0.012 to about 11.11111 eight / 1112, more preferably about 0.026 to about 5 55 111 eight / 1112. 5 The applied current density can be used in the range of about 10 to about 2,000 mA / m2. In another embodiment of the invention, the applied current is generated in a range between about 50 to about 600 mA / m2. In a further specific example of the present invention, the EF applied current is generated in a range between about 60 to about 100 mA / m2. Table 11 provides a better set of parameters for treating disorders and conditions. Table 11 provides the particular disorder, condition, organ or system to which this parameter group was administered. Table 11 also provides special parameter values, however it is understood that the values are approximate and equal ranges are contemplated by the present invention. 67 200423986 Exposure time 4 minutes 2 minutes 24 hours / day, 7 days 2 and 24 hours / day 2 hours / day, 56 days 24 hours 30 minutes and 24 hours of applied current density (in mA / m2) 60 , 200, 600, or 2,000 2000 10, 50, and 100 60 or 600 l ^ r S and T? ^ G 0.42 0.026-0.32 0.42 EF voltage (in volts) 2000 3000 ί_ EF frequency (in Hertz) ) SS 50 (30 kV / m) 50 (30 kV / m) Obstacles, pathologies, organs or systems 撵 mw toward · CN Xisu: mw > Λ a US: dysfunction related to fibroblast proliferation ass Λ a House rheumatoid arthritis A Wenzhai A meal j, 铼 parameter group rH (N inch ο 200423986 Royal H < l > order 〇Oh l〇o rH 0.035-0.5 (N inch 〇7000 3000 »ε /« SS ° 9 co σ / —s 〇 \ 〇 '^ ― 1 \ χ > ° ^ ν VO > -tfrC mr ^ r + (N cd + CN σ3 + (N cd + (N cd + C4 cd 0 UUU υ u € ε稳定 溪 镩 食 镓 溪 灿 璨 撵 4 ti: broken " S 铼: S) Chu Zhu: s Zhu forget 00 〇 \ 〇rH rH τ-Η (N cn inch τ-Η 200423986 30 minutes and 24 hours 12 minutes 1 hour / day, 72 times or 100 days 7 times with 7000 V; 23 times with 30000 V Φ # ⑺〇, compared with 褽 · 螫. 2 4 1, ^ ^ ^ ^ ~ S60 or 600 0.0001-0.42 2.3-11.1 7.5-11.1 50 and 15000 (AC, DC +? DC-) 9000 or 30,000 30000 < Increases the induction of ConA by immune system cells ConA increases the induction of ConA by immune system cells is related to electrolyte imbalance Obstacle * 2 * 2 Test 1 2 2 U U 2 two targets w (55 2 watch 1 wear > 〇〇 fatigue in 00 〇 \ 200423986

cn minutes / periods, every 14 days (N 〇® ^ CN ^ T called #s inin Φ 你 〇yo ^ S? ό 〇 > cn ^ 1 O m ^ 0.08-1.12 3.75-5.55 ( N rH rH 1 od 0.024-1.12 8000 15000 40000 < 50 (12-40 kV / m) 50 (12-40 kV / m) Stress response and cytokine-induced disorders and electrolyte imbalance disorders Suppression Cell proliferation disorder (N CN CO CM 200423986) The present invention is also directed to a method for determining a desired parameter set (such as EF characteristics, induced current density, applied current density, and exposure time), thereby maximizing The desired effect is obtained in a biological test subject. In a preferred embodiment of the present invention, the optimization method includes the following 5 steps: confirming a desired one to be induced in an organism or part thereof Biological effect (for example, causing inward ma ion flow in muscle cells); choose a value as an average applied current density or an induced current density in the cell membrane of the organism or part thereof, where the applied current is In terms of this number It preferably falls within a range of about 10 mA / m2 to about 2,000 mA / m2, and falls within a range of about 0.00lniA / m2 to about 15 mA / m2 in terms of induced current; A value of applied current or EF (such as frequency and EF voltage) other than the selected current density is generated; a discontinuous time period is selected to generate the applied current density, where the period falls between about 2 minutes to continuous or Within a discontinuous range of about 10.80 minutes; applying the applied current or EF 15 to produce the selected current density; determining the extent to which the desired biological effect occurs; and repeating the steps Either of these. Preferably, the optimization step is also accompanied by generating a dose-response curve as a function of a selected value. In another preferred embodiment, the value of the applied current or EF is It is determined in view of the body shape, body weight, body fat percentage of the organism, and 20 other factors related to the induction current throughout the cell membrane. In some specific examples of the present invention, it is used to regulate the cell membrane across the body in vivo. The parameters of the ion current are The combinations presented in Table 12 are exemplified. In other specific examples of the present invention, the parameters used to regulate the ion flux across the cell membrane in vitro are exemplified by the combination 25 presented in Table 13. 72 200423986 Table 12-Exemplary parameters for adjusting ion current in vivo Parameter group EF voltage (in volts) EF frequency (in Hz) Induced current density (in mA / m2) Applied current density (In mA / m2) Exposure time 1 2,000 50 0.026-0.32 2 hours / day for 7 days 2 2,000 50 0.026-0.32 2 hours / day for 56 days 3 7,000 50 (17.5 KV / m) 0.035-0.5 60 minutes 4 30,000 60 7.5-11.1 30 minutes 5 7,700 50 0.015-0.22 2 hours / day, 6 days / week, 15 weeks 6 15,000 60 3.8-5.6 20 minutes / day, 4 times per period, 15 days 7 50 50 0.0001-0.42 72 days 8 15,000 50 0.0001-0.42 100 days 9 3,000 60 0.006-0.08 35 days 10 10,000 60 0.05-0.7 15 minutes / day, which lasts 91 days 11 7,000 60 (17.5 KV / m) 0.035-0.5 15 minutes / Day, 7 days 12 8,000 40 KV / m 2 hours 13 15,000 50 3.75-5.55 30 minutes / period, every other day, 2 weeks 14 10,000 -30,000 50 2.5-11.1 30 minutes 15 30,000 50 7.5-11.1 15 minutes / day, 3 times / week, 2 weeks 16 30,000 50 7.5-11.1 30 minutes / day 17 30,000 60 7.5-11.1 30 minutes / day 73 200423986 18 2,400 50 (6 KV / m) 0.012-0.17 19 8,000 50 (40 KV / m) 0.08-1.12 2 hours 20 1,200 50 (6 KV / m) 0.012-0.17 1 hour / day, 7 days 21 50 (12-40 KV / m) 0.024-1.12 30-120 minutes / day, 4 weeks 22 50 (12-40 KV / m) 0.024- 1.12 30-120 minutes / day over 8 weeks 23 2,400 50 (6 KV / m) 0.012-0.17 30 minutes 24 2,400 50 (6 KV / m) 0.012-0.17 120 minutes 25 10,000 '20,000 or 30,000 2.5-11.1 20 minutes 26 10,000 2.5-3.7 10 minutes / day, 3 times / week, 5 weeks 74 200423986 Table 13_ Model parameters for adjusting ion current in vitro Parameter group EF voltage (in volts) EF frequency (in Hz) Unit) Induced current density (in mA / m2) Applied current density (in mA / m2) Exposure time 1 60 60 4 minutes 2 60 200 4 minutes 3 60 600 4 minutes 4 60 2000 4 minutes 5 60 2000 4 minutes 6 60 10 24 hours / day, 7 days 7 60 50 24 hours / day, 7 days 8 60 100 24 hours / day, 7 hours Day 9 50 (30 KV / m) 0.42 2 hours 10 50 (30 KV / m) 0.42 24 hours 11 50 (30 KV / m) 0.42 24 hours 12 60 60 or 600 30 minutes 13 60 60 or 600 24 hours 14 60 60 12 minutes 15 60 60 4 minutes 16 3,000 50 (30 KV / m) 0.42 24 hours 17 50 100-1000 18 50 10 7 days 19 50 50 7 days 20 50 100 7 days 21 15,000 60 22 1,000 50 (150 KV / m) 3.9 48 hours 23 1,000 50 (10 KV / m) 0.26-0.34 48 hours 24 50 (8.3 KV / m) 0.28 48 hours 75 200423986 In an alternative embodiment, the present invention can be used as a diagnostic tool Determine if a body is experiencing a particular obstacle or condition. Specific parameters related to the prevention, improvement, and treatment of an obstacle or condition can be used to detect the presence of the same obstacle or condition. These parameters can be used as a diagnostic tool and can be used to monitor response. If the patient does not respond to a given parameter set (the parameter set is related to the disease), then a lack of a response implies that the patient has not experienced a particular disorder or condition. Alternatively, if the patient responds to a given parameter set (the parameter set is related to the disease), then the presence of a response indicates the presence of a particular disorder and / or condition. Specific diagnostic examples of the present invention can be used for each disorder and / or condition (for each, a specific EF parameter set has been determined). It will be clear that the present invention may be applied differently from the method particularly described in the foregoing description and embodiments. Many modifications and variations of the present invention are possible based on the above teachings, and therefore, fall within the scope of the patent application attached to the document. The entire disclosure of each document (including patents, patent applications, journal articles, excerpts, laboratory manuals, books, or other disclosures) cited in the background, detailed description, and examples is hereby incorporated in its entirety. Incorporated into this case for reference. The exact electrotherapeutic device and method of applying an electric field are disclosed in U.S. Patent Application Serial No. 10 / 017,105 (filed on December U. 2001), which U.S. patent application is hereby incorporated in its entirety as this Spring test information. 76 200423986 [Simplified figure of the month j Figure 1 shows a field exposure dish in an EF exposure system; Figure 2 shows the percentage of viable cells after EF exposure; Figure 3 shows In both EF-exposed and unexposed cell suspensions containing 12.5 pg / ml Con-A, there was a significant increase in cell numbers at high [(:: ^-| .; Figures 4A and 4B summarize Conon A with different concentrations, with and without ImM CaCl2, EF exposed cell cultures (cen cuitures); 10 Figure 5 shows EF exposure to chromium containing phytohemaglutinin (PHA) There was a significant increase in high [ca2 +] c cells in both cells with and without exposure; Figure 6 shows that when supplemented with Con-A at 3.125-12.5 gg / ml, whether exposed to EF or not The exposed cells showed a significant increase in high [Ca2 +] c cells compared to those stimulated with 15.025 pg / ml Con-A; Figure 7 illustrates an example in splenocytes (Spien0Cyte cells), the increase in calcium ion concentration induced by coonA; Figure 8 shows the Time course of DiBAC staining intensity in embryonic cells of 20BALB3T3 mice stimulated with 0.4 μM A23187; Figure 9 shows an electric field that produces a current density of about 200 μA / μ2 at 100 Hz ( 1EF) on the membrane potential in BALB 3T3; Figure 10 also shows the effect of an electric field (EF) that produces a current density of about 200μA / (: 2η 2 at 1000Hz) on the membrane potential in BALB 3T3 77 200423986 Figure 11 shows the effect of stress on the level of plasma adrenocorticotropic hormone (hereinafter referred to as "ACTH"); Figures 12A and 12B show exposure to EF for normal (A) and ovariectomized ) (B) The effect of plasma ACTH levels in rats; 5 Figure 13 shows the effect of EF exposure on blood poly ACTH levels in normal rats (n == 6); Figures 14A and 14B show Effect of EF exposure on changes in about East-induced-induced plasma glucose levels in normal (A) and indo-removed (B) rats; Figures 15A and 15B show EF exposure for normal ( (A) Constrained-induced plasma lactate level in rats with ovariectomy (B) Figure 16 shows the effect of EF exposure on confinement _ induced plasma pyruvate levels in ovariectomized rats; Figure 17 shows the effect of EF exposure on confinement _ 15 induction in ovariectomized rats Effect of WBC count; Figure 18 illustrates an example of the concept of the electric field generated by the use of an EF therapy device (in this case, a BiOniTrón chair from the White Brothers Health Science Association) Outline; Figure 19 is a schematic 20 diagram of a preferred treatment device of the present invention; Figures 20A and 20B show another preferred EF therapy device; Figures 21A and 21B show another The preferred EF therapy device; Figure 22 is a diagram of the preferred electronic configuration of the -effect tf out-EF therapy device, 78 200423986 Figure 23A is a front view of a stimulated human body, Figure 23B The diagram is a perspective view, and FIG. 23C is a diagram showing an EF measurement sensor attached to the neck of the body; FIG. 24 shows a device for measuring the EF therapy device. The device that produces the sense of 5 current; Figure 25 Shows the relationship between an applied voltage and an induced current; Figure 26 shows the relationship between the position of a head electrode and the current induced in the neck; 10 Figure 27 illustrates an example Induced current density (mA / m2) at different locations of a grounded human individual; and Figure 28 shows the mitigation effect of EF exposure on different human symptoms. [Representation of the main components of the diagram]

79

Claims (1)

200423986 The scope of patent application: 1. A method for treating or preventing an obstacle that causes an abnormal ion concentration in a cell of an organism or a part thereof or is caused by the abnormal ion concentration. The method It includes restoring the cells to a normal ion concentration, and includes applying an external electric field to the organism or part, the external electric field generating a cell membrane of the cells of about 0.001 mA / m2 to about 15 mA / m2 average induced current density. 2. The method of claim 1 in which the ions include calcium ions. 10 3. The method of claim 1, further comprising providing the organism or a part thereof with a calcium supplement, a vitamin D supplement, a phytolectin supplement, or a combination of these supplements . 4. The method of claim 3, wherein the lectin supplement is provided and the lectin supplement comprises concanavalin A 15 or wheat germ agglutinin. 5. The method according to any one of claims 1 to 4, wherein the average induced current density is about 0.01 mA / m2 to about 2 mA / m2. 6. The method of claim 5, wherein the biological system is a human, and the electric field generates the average sensation on the cell membrane of the human cell. The current density should last from about 10 minutes to about 240 minutes. Continuous period. 7. The method of claim 6, further comprising the subsequent repeated application of the external electric field to the human or part thereof and the repeated generation of the average induced current density for an additional 80 200423986 continuous period . 8.-A device for performing the method as described in the first item of the patent application, wherein the device is an electric field therapy device, the device comprises: (a)-a main electrode and an opposite electrode; 5 (b)-used A voltage generator that applies a voltage to the electrodes; (c) an induced current generator that is controlled by changing the voltage or the phase. The distance between the counter electrode and the organism or part thereof The external electric field; and φ (d) —a power source for driving the voltage generator. 10 9. The device according to item 8 of the application, wherein the main electrode is not in contact with the living body or a part thereof. 10. A method for treating a proliferative cell disorder, the method comprising altering an ion flow across a cell membrane of an organism or part thereof, and including applying an external electric field to the organism or part, the external electric field being applied to the The cell 15 membrane produces an average induced current density of about 0.1 mA / m2 to about 2 mA / m2. · 11. The method of claim 10, wherein the average induced current density is about 0.2 mA / m2 to about 1.2 mA / m2. ^ 12. The method according to item 11 of the patent application range, wherein the average induced current body 20 has a density of about 0.29 mA / m2 to about 1.12 mA / m2. 13. The method of claim 11 in which the ions include calcium ions. 14. The method of claim 10, further comprising providing the organism or a part thereof with a calcium supplement, a vitamin D supplement, a 81 200423986 plant lectin supplement, or one of the supplements combination. 15. The method of claim 14 in which the phytolectin supplement is provided, and the phytolectin supplement comprises concanavalin A or wheat germ agglutinin. 5 16. The method of claim 11 in which the proliferative cell disorder involves differentiated fibroblasts. 17. The method of claim 11 or 14, wherein the biological system is a human, and the electric field generates the average induced current density on the cell membrane of the human cell for about 10 minutes to about 240 minutes. 10 consecutive minutes. 18. The method of claim 17 further comprising the additional continuous period of subsequent repeated application of the electric field to the human or part thereof and repeated generation of the average induced current density for approximately 30 minutes to approximately 90 minutes. 15 19. The method of claim 18, wherein the human is arranged in a hospital or clinic bed. 20.-A device for performing the method according to item 11 of the patent application, wherein the device is an electric field therapy device comprising: (a)-a main electrode and an opposite electrode; 20 (b)-for A voltage generator that applies a voltage to the electrodes; (C) an induced current generator that controls the external electric field by changing the voltage or the distance between the opposing electrode and the organism or part thereof; And (d)-a power source for driving the voltage generator. 82 200423986 21. The device according to claim 20, wherein the main electrode is not in contact with the living body or a part thereof. 22.-A method for treating an electrolyte imbalance, the method comprising altering an ion current across a cell membrane of an organism or part thereof, and comprising applying an external electric field to the living body or part, the external electric field being applied to the cell membranes This produces an average induced current density of about 0.4 mA / m2 to about 6.0 mA / m2. 23. The method of claim 22, wherein the average induced current density is about 0.4 mA / m2 to about 5.6 mA / m2. 10 24. The method of claim 23, wherein the average induced current density is about 0.43 mA / m2 to about 5.55 mA / m2. 25. The method of claim 23, wherein the ions include calcium ions. 26. The method of claim 22, further comprising providing a calcium supplement, a vitamin D supplement, a phytolectin supplement, or a combination of these supplements to the 15 biological objects or parts thereof. . 27. The method of claim 26, wherein the plant lectin supplement is provided and the plant lectin supplement comprises concanavalin A or wheat germ agglutinin. 20 28. The method of claim 23 or 26, wherein the biological system is a human, and the electric field generates the average induced current density on the cell membrane of the human cell for about 10 minutes to about 240. Continuous period of minutes. 29. The method of claim 28, which further comprises an additional continuous period of subsequent repeated application of the electric field to 83 200423986 the human or part thereof and the repeated generation of the average induced current density for approximately 30 minutes to approximately 90 minutes. 30. The method according to item 29 of the patent application, wherein the human is arranged in a hospital or clinic bed. π 31. A device for performing the method as described in the scope of patent application No. 22, wherein the device is an electric field therapy device including the following: (a) —the main electrode and an opposite electrode; Xin (b) — A voltage generator for applying a voltage to the electrodes; 10 (c) an induced current generator that controls the voltage by changing the voltage or the distance between the opposite electrode and the organism or part thereof External electric field; and (d)-a power source to drive the voltage generator. 32. The device according to item 31 of the patent application, wherein 15 of the main electrode is not in contact with the living body or a part thereof. 33. A method for treating a disorder related to serum calcium concentration, the method comprising altering a mother ion current across a cell membrane of an organism or part thereof, and including applying an external electric field to the organism or part, the external An electric field produces an average induced current density on the cell membrane of approximately 0.3 mA / m2 to approximately 0.6 mA at 20 mA / m2. 34. The method of claim 33, wherein the average induced current density is about 0.4 mA / m2 to about 0.5 mA / m2. 35. The method of claim 34, wherein the average induced current density is about 0.42 mA / m2. 84 200423986 36. The method according to claim 33, further comprising providing the organism or a part thereof with a calcium supplement, a vitamin D supplement, a phytolectin supplement, or one of the supplements combination. 37. The method of claim 36, wherein the plant lectin supplement is provided, and the plant lectin supplement comprises concanavalin A or wheat germ agglutinin. 38. The method of claim 34 or 36, wherein the biological system is a human, and the electric field generates the average induced current density on the cell membrane of the human cell for about 10 minutes to about 240 minutes 10 consecutive minutes. 39. The method of claim 38, further comprising an additional continuous period of subsequent repeated application of the electric field to the human or part thereof and repeated generation of the average induced current density for approximately 30 minutes to approximately 90 minutes. 15 40. The method of claim 39, wherein the human is arranged in a hospital or clinic bed. 41. A device for performing the method as described in claim 33, wherein the device is an electric field therapy device comprising: (a)-a main electrode and an opposite electrode; 20 (b)-for applying A voltage generator from a voltage to the electrodes; (c) an induced current generator that controls the external electric field by changing the voltage or the distance between the opposing electrode and the organism or part thereof; and (d)-the power source used to drive the voltage generator. 85 200423986 42. The device according to claim 41, wherein the main electrode is not in contact with the living body or a part thereof. 43 · —A method for lowering ACTH or cortisol level, which comprises changing the ion M across a cell membrane of an organism or part thereof and including applying an external electric field to the organism or part The external electric field generates an average induced current density on the cell membranes of about 0.000 mA / m2 to about 12 mA / m2. 44. The method of claim 43 in the patent application range, wherein the average induced current density is about 0.035 mA / m2 to about 1 L1 mA / m2. 10 45. The method of claim 44 wherein the average induced current density is about 0.035 to about 0.5 mA / m2. 46. The method according to item 43 of the patent application, wherein the ions include calcium ions, and the method further includes providing a supplement to the organism or a part thereof, a vitamin D supplement, and a lectin supplement 15 Or a combination of these supplements. 47. The method of claim 46, wherein the phytoalexin supplement is provided ' and the phytoalexin supplement comprises concanavalin A or wheat germ agglutinin. 48. The method of claim 44 < 46, wherein the biological system 20 is a human, and the electric field generates the average induced current density on the cell membrane of the human cell for about 10 minutes Continuous period to approximately ㈣ minutes. 49. The method of claim 48, further comprising the subsequent repeated application of the electric field to the human or part thereof and the repeated generation of the average 86 200423986 induced current density for an additional continuous period of about 30 minutes to about 90 minutes. 50. The method of claim 49, wherein the human is arranged in a hospital or clinic bed. 5 51. —A device for performing the method as claimed in the scope of patent application No. 43, wherein the device is an electric field therapy device including the following: (a) —main electrode and an opposite electrode; (b) —for A voltage generator that applies a voltage to the electrodes; (c) an inductive current generator that controls the outside by changing the voltage or the distance between 10 pairs of electrodes in the phase and the organism or part thereof Electric field; and (d)-a power source to drive the voltage generator. 52. The device of claim 51, wherein the main electrode is not in contact with the living body or a part thereof. 15 53. A method for treating stress, the method comprising altering an ion current across a cell membrane of an organism or part thereof, and comprising applying an external electric field to the organism or part, the external electric field being generated on the cell membrane One is an average induced current density of about 0.03 mA / m2 to about 12 mA / m2. 54. The method of claim 53 in which the average induced current density is about 0.035 mA / m2 to about 11.1 mA / m2. 55. The method of claim 54 in which the ions include calcium ions. 56. The method of claim 53, further comprising providing the organism or a part thereof with a calcium supplement, a vitamin D supplement, a 87 200423986 phytolectin supplement, or one of the supplements combination. 57. The method of claim 56 in which the plant lectin supplement is provided and the plant lectin supplement comprises concanavalin A or wheat germ agglutinin. 5 58. The method of claim 54 or 56, wherein the biological system 11 is a human, and the electric field generates the on the cell membrane of the human cell. The average induced current density lasts about 10 minutes to Continuous period of about 240 minutes. φ 59. The method according to item 58 of the scope of patent application, further comprising an additional continuous period of subsequent repeated application of the electric field to the human or part thereof and repeated generation of the average induced current density for approximately 30 minutes to approximately 90 minutes. 60. The method of claim 59, wherein the human is arranged on a hospital or clinic bed. 15 61. A device for performing the method as claimed in the scope of patent application No. 53, wherein the device is an electric field therapy device including the following: H (a)-the main electrode and an opposite electrode; (b)-used A voltage generator that applies a voltage to the electrodes; (c) an induced current generator that changes the voltage or the distance between the phase ψ 20 pair of electrodes and the organism or part thereof Controlling the external electric field; and (d)-a power source for driving the voltage generator. 62. The device of claim 61, wherein the main electrode is not in contact with the living body or a part thereof. 88 200423986 63.-A method for treating arthritis, the method comprising changing the ion current across a cell membrane of an organism or part thereof, and including applying an external electric field to the organism or part, the external electric field being applied to the cell membranes This produces an average induced current density of about 5 degrees from about 0.02 mA / m2 to about 0.4 mA / m2. 64. The method of claim 63, wherein the average induced current density is about 0.025 mA / m2 to about 0.35 mA / m2. 65. The method of claim 64, wherein the average induced current density is about 0.026 mA / m2 to about 0.32 mA / m2. 10 66. The method of claim 64, wherein the ions include calcium ions. 67. The method of claim 63, further comprising providing the living body or a part thereof with a calcium supplement, a vitamin D supplement, a phytolectin supplement, or a combination of these supplements. 15 68. The method of claim 67, wherein the phytohemagglutinin supplement is provided and the phytohemagglutinin supplement comprises concanavalin A or wheat germ agglutinin. 69. The method of claim 64 or 67, wherein the biological system is a human, and the electric field generates the 20 average induced current density on the cell membrane of the human cell for about 10 minutes to about 240. Continuous period of minutes. 70. The method of claim 69, further comprising an additional 89 200423986 continuous period of subsequent repeated application of the electric field to the human or part thereof and repeated generation of the average induced current density for approximately 30 minutes to approximately 90 minutes. 71. The method of claim 70, wherein the human is arranged in a hospital or clinic bed. 72.-a device for performing the method according to item 63 of the scope of patent application, 5 wherein the device is an electric field therapy device comprising: (a)-a main electrode and an opposite electrode; (b)-for A voltage generator that applies a voltage to the electrodes; (c) an induced current generator that controls the external 10 electric field by changing the voltage or the distance between the opposite electrode and the organism or part thereof And (d)-the power source used to drive the voltage generator. 73. The device of claim 72, wherein the main electrode is not in contact with the organism or part thereof. 74. A method for treating overweight, which method comprises changing the ion current across a cell membrane of a 15 organism or part thereof, and comprising applying an external electric field to the organism or part, the external electric field being applied to the cell membrane This produces an average induced current density of about 0.02 mA / m2 to about 1.5 mA / m2. 75. The method of claim 74, wherein the average induced current 20 has a density of about 0.02 mA / m2 to about 1.2 mA / m2. 76. The method of claim 75, wherein the average induced current density is about 0.024 mA / m2 to about 1.12 mA / m2. 77. The method of claim 75, wherein the ions include calcium ions. 90 200423986 78. The method of claim 74, further comprising providing the organism or a part thereof with a calcium supplement, a vitamin D supplement, a phytolectin supplement, or one of the supplements combination. 79. The method of claim 78, wherein the plant lectin supplement is provided and the plant lectin supplement comprises concanavalin A or wheat germ agglutinin. 80. The method of claim 74 or 78, wherein the biological system is a human, and the electric field generates the average induced current density on the cell membrane of the human cell for about 10 minutes to about 240 minutes 10 consecutive minutes. 81. The method of claim 80, further comprising an additional continuous period of subsequent subsequent application of the electric field to the human or part thereof and repeated generation of the average induced current density for approximately 30 minutes to approximately 90 minutes. 15 82. The method of claim 81, wherein the human is arranged in a hospital or clinic bed. 83.-a device for performing the method as claimed in the scope of patent application 74, wherein the device is an electric field therapy device comprising: (a)-the main electrode and an opposite electrode; 20 (b)-for A voltage generator that applies a voltage to the electrodes; (c) an induced current generator that controls the external electric field by changing the voltage or the distance between the opposing electrode and the organism or part thereof; And (d)-a power source for driving the voltage generator. 91 200423986 84. The device according to item 83 of the patent application, wherein the main electrode is not in contact with the organism or a part thereof. 85. A method for determining the optimal parameters for treating an electric field exposure outside a disorder, the method comprising: (i) identifying a desired biological response to be induced in a living organism, (ii ) Select or measure an average induced current density on the cell membrane of the organism or a tissue sample or cell derived from the organism; (iii) Select or measure an external electric field, the external electric field A screened or measured induced current density is generated at a specific distance from the organism, sample or culture; (iv) selecting or measuring a screened or measured induced current density on the cell membrane 15 (v) applying the selected or measured electric field to the organism, sample, or culture, so that the selected or measured induced current density on the cell membranes should be screened or measured over time (Vi) determine the extent to which the desired biological response occurs; 20 (vii) selectively repeat any of steps (ii) to (vi); and / or (viii) confirm the best Geo-induced The desired biological response of selected Examples of the induced current or the measured density value, as a value of the selected external electric field or the measured value or as the continuous time period is measured or screened. 92 200423986 86. The method of claim 85, further comprising, before step (viii), generating a dose-response curve as the selected or measured induced current density, the selected or measured external electric field, or A function of a continuous time period that is selected or measured. 5 87. The method of claim 85, further comprising before step k (viii), selecting or measuring the following: the number of times that step (v) was repeated; between the repetitions of step (V) Time interval; and φ the selected or measured induced current density is generated over the entire period of 10 times on the cell membrane. 88. The method of claim 85, wherein the selected or measured induced current density is about 0.001 mA / m2 to about 15 mA / m2. 89. The method of claim 85, wherein the cell lines are in a culture. 15 90. The method of claim 89, wherein the cell lines in the culture are human cells. 91. The method of claim 85, wherein the cell lines are located in a living organism or part thereof. β 92. The method of claim 91, wherein the living biological system is t 20 -human. 93. The method of claim 85, wherein the induced current density is obtained by measuring the induced current flowing in a given area of the living organism or a part thereof, and converting the measured current into A voltage signal is selected or measured by converting the voltage signal into an optical signal and then converting the optical signal into a voltage signal and analyzing the waveform and frequency by 93 200423986. 94. The method of claim 85, wherein the induced current density is represented by J, and J is represented by J = I / B. 5 95. The method of claim 85, further comprising providing the organism, sample, or culture with a calcium supplement, a vitamin D supplement, a phytolectin supplement, or the supplement. A combination. 96. The method of claim 95, wherein the phytolectin supplement is provided and the phytolectin supplement comprises concanavalin 10A or wheat germ lectin. 97.-a device for performing the method according to the scope of patent application No. 85, wherein the device is an electric field therapy device comprising: (a)-the main electrode and an opposite electrode; (b)-for applying A voltage generator to the electrodes; 15 (c) an induced current generator to control the external electric field by changing the voltage or the distance between the opposing electrode and the organism or part thereof; And (d)-a power source for driving the voltage generator. 98. For a device according to item 97 of the application, wherein no 20 of the main electrode contacts the organism or a part thereof. 99. A method for treating a proliferative cell disorder, the method comprising altering an ion current across a cell membrane of an organism or part thereof, and comprising contacting the organism or part with an electric current, the current being on the cell membrane This results in an average applied current density of about 2004 mA to about 100 mA / m2 to about 100 mA / m2. 100. The method of claim 99, wherein the ions include calcium ions, and the average applied current density is generated on the cell membranes for a continuous period of substantially at least about 7 days. 5 101. The method of claim 99, further comprising providing the organism or a part thereof with a calcium supplement, a vitamin D supplement, a phytolectin supplement, or a combination of these supplements . 102. The method of claim 101, wherein the plant lectin supplement is provided, and the plant lectin supplement comprises concanavalin 10 A or wheat germ agglutinin. 103. The method according to claim 99, 100 or 101, wherein the biological system is a human. 104. A current therapy device for performing a method as claimed in claim 99. 15 105. —A method for treating a stress-related disorder or condition, the method comprising altering an ion current across a cell membrane of an organism or part thereof, and including contacting the organism or part with an electric current, the electric current being The cell membranes have an average applied current density of about 60 mA / m2 to about 600 mA / m2. 20 106. The method of claim 105, wherein the ions include calcium ions. 107. The method of claim 105, further comprising providing the living body or a part thereof with a calcium supplement, a vitamin D supplement, a phytolectin supplement, or a combination of these supplements. 95 200423986 108. The method as claimed in claim 107, wherein the phytolectin supplement is provided and the phytolectin supplement comprises concanavalin A or wheat germ agglutinin. 109. The method of claim 105 or 107, wherein the organism 5 is a human. 110. An electrotherapeutic device for performing a method such as that described in claim 105. 111. A method for treating a disorder related to serum calcium concentration, the method comprising altering a flow of calcium ions 10 across a cell membrane of an organism or part thereof and including contacting the organism or part with an electric current, the electric current An average applied current density of about 60 mA / m2 to about 2000 mA / m2 is generated on the cell membranes. 112. The method of claim 111, wherein the ions include calcium ions. 15 113. The method according to claim 111, further comprising providing the organism or a part thereof with a calcium supplement, a vitamin D supplement, a phytolectin supplement, or a combination of these supplements . 114. The method of claim 113, wherein the phytolectin supplement is provided and the phytolectin supplement contains concanavalin 20 A or wheat germ lectin. 115. The method of claim 111 or 113, wherein the organism is a human. 116. The method of claim 111, wherein the current generates the average applied current density on the cell membranes for a continuous period of about 1 minute 96 200423986 to about 20 minutes. 117. The method of claim 116, wherein the current generates the average applied current density on the cell membranes for a continuous period of about 2 minutes to about 10 minutes. 5 118. An electrotherapy device for performing a method such as that claimed in claim 111. · 119. A method for determining the optimal parameters of current exposure for treating a disorder, the method comprising: _ (i) identifying a desired biological response to be induced in a living organism; (ii) selecting or measuring an average induced current density on the cell membrane of the organism or a cell derived from a tissue sample or culture of the organism, wherein the average applied current density is about 10 mA / m2 to about 2,000 mA / m2; 15 (iii) selecting or measuring a current that will produce the selected or measured applied current density; _ (iv) selecting or measuring a current used to produce the selected or measured Continuous time period during which the current density is applied; (v) applying the selected or measured current to produce the selected or measured applied current density for the continuous time period during which the selected or measurement is made; (vi) Determine the degree to which the desired biological response occurs; (vii) repeat any of steps (ii) to (vi) to generate a dose-response curve as the selected or measured current, the selected 97 200423986 OR A function of the applied current density or a continuous time period of the selection or measurement; and (viii) confirming that the selected or measured current, the selected or measured current that best induces the desired biological response The applied current density or 5 is the value during the continuous time that is selected or measured. 120. The method of claiming scope 119, further comprising, before step (viii), selecting or measuring the following: The number of times that step (v) is repeated; the time interval between the repetitions of step (v); and 10 the entire period during which the selected or measured induced current density is generated on the cell membrane. 121. The method of claim 120, wherein the cell lines are located in a culture. 122. The method of claim 121, wherein the cell lines in the culture 15 are human cells. 123. The method of claim 120, wherein the cell lines are located in a living organism or a part thereof. 124. The method of claim 123, wherein the living biological system is a human. 20 125. The method of claim 120, further comprising providing the organism, sample, or culture with a calcium supplement, a vitamin D supplement, a phytolectin supplement, or a supplement of the supplement. A combination. 126. The method of claim 125, wherein the phytolectin 98 200423986 supplement is provided, and the phytolectin supplement comprises concanavalin A or wheat germ lectin. 127. An electrotherapy device for performing a method such as that described in claim 120. 99