JP2009125549A - Living tissue normalization apparatus and treatment device - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a living tissue normalization apparatus and so on to activate protein at the most effective voltage value and rising time by a method to activate a normalization mechanism of a living body or living tissue via protein by adding a feeble electric current and heat. <P>SOLUTION: In the living tissue normalization apparatus to activate a normalization mechanism of a living body or living tissue via protein by electrifying the living body or living tissue with feeble direct current generated by applying direct voltage intermittently at a predetermined interval and adding heat to the living body or living tissue, pulse waveforms of input voltage to be applied are square waves, a period of time to reach a rising peak value in one cycle of the pulse waves is between 0.05 millisecond and 0.2 millisecond (both inclusive) and the peak value is between 3.0 V and 20.0 V (both inclusive). <P>COPYRIGHT: (C)2009,JPO&INPIT

Description

  The present invention relates to a normal tissue normalizing apparatus that activates a living body or a living tissue, and more particularly to a normal tissue normalizing apparatus that activates a normalization mechanism of the living body or the living tissue.

  In recent years, ubiquitin present in every cell has a function that has been elucidated and binds to a protein that is no longer needed to serve as a landmark. Unnecessary protein (ubiquitinated protein) to which this ubiquitin is bound is incorporated into the enzyme proteasome and decomposed using this ubiquitin as a mark. Such a biological normalization mechanism for decomposing unnecessary proteins works as a ubiquitin / proteasome system and is involved in normalization of many tissues such as cell division, DNA repair, protein quality control, and immunity.

  In addition, the living body normalization mechanism is activated by heating a predetermined part of the living body and inducing the expression of heat shock protein (hereinafter referred to as “HSP”) with an electric thermotherapy device.

  The HSP refers to a group of proteins having molecular weights of tens of thousands to about 150,000, also called stress proteins, and is classified into several families according to molecular weights. HSP binds non-covalently to the hydrophobic part of nascent proteins, denatured proteins and abnormal proteins, assists in protein folding, transport to intracellular organelles, refolding and degradation of denatured proteins, and quality of intracellular proteins Management is performed to prevent accumulation of abnormal or denatured proteins in the cell. These functions are collectively called molecular chaperones, and HSP is induced by various physical and chemical injury factors including heat shock. It is an established fact that cells that express a large amount of HSP acquire a strong resistance against various damaging factors.

  HSP70 having a molecular weight of 72 kilodaltons belonging to the HSP70 family is a protein that is induced for the first time by stress, and the research is most advanced. If HSP70 is pre-overexpressed in advance by exposing the cell to non-lethal stress such as heat shock, the cell can survive and exhibit strong resistance to a lethal injury factor.

  This resistance not only prevents the accumulation of abnormal and denatured proteins in the cell through the function of molecular chaperones, but also preserves the function of intracellular organelles such as mitochondria in stressed cells, It has been shown to suppress cell necrosis to suppress inflammatory responses and to suppress apoptosis to suppress cell loss (Samali, A. et ai., Cell Stress & Chaperones 3: 228, 1998).

  In various pathological conditions, cells are exposed to physical and chemical stress. In many disease models using experimental animals, it has been clarified that HSP70 is excessively expressed in some way, and injury is reduced, and expectations for clinical application of HSP70 are increasing (Minowada, G et al., J. Clin. Invest., 95: 2, 1995 and references cited therein).

  When referring to the various stresses that cells encounter in the context of human disease, a typical stress is ischemia. When the experimental animal is preliminarily subjected to a heat shock load and HSP70 is overexpressed in advance, even when the cerebral artery or coronary artery is ligated, the brain (Kitagawa, K. et al., J. Cereb. Blood Flow Metab., 11: 449, 1991) and heart (Donnelly, TJ et al., Circulation 85: 1048, 1992) have been shown to shrink. In addition, the effect of suppressing myocardial infarction has also been shown in mice in which HSP70 gene has been introduced (Maber, M. S. et al., J. Clin. Invest., 95: 1446, 1995). The inhibitory effect of HSP70 on cell damage due to ischemia is applied not only to the brain and heart but also to all organs.

  Reactive oxygen and free radicals increase in production with infection, inflammation, degenerative diseases, autoimmune diseases, arteriosclerosis, and aging, causing cell damage. HSP70 has been shown to suppress cell damage caused by these active oxygen and free radicals (Polla B. S. et al., Proc. Natl. Acad Sci. USA, 93: 6458, 1996).

  For ischemia / reperfusion injury, increased production of active oxygen during reperfusion is considered as one of the major etiologies, and it is known that HSP is reduced in the brain, heart, liver, small intestine, etc. . This protective action by HSP is applied to all organs. Organ transplantation is a typical example of ischemia-reperfusion injury. In fact, it has been shown that over-expression of HSP70 improves the survival rate of skin grafts (Koenig, WJ et al., Plast Recontsr. Surg., 90: 659, 1992). It has also been reported that acute rejection decreases as the transplanted liver expresses more HSP70 during transplantation (Flohe, S. et al., Traspl. Lnt., 11:89, 1998). Ultraviolet rays, radiation, heavy metals, alcohol, anticancer agents and paraquat cause injury mainly due to active oxygen and free radicals. HSP70 can also be expected to prevent and treat skin, mucous membrane, eye lens and retina damage caused by ultraviolet rays and radiation, and also to treat alcoholic organ damage, heavy metals and drug addiction.

  In addition, cancer cells express HSP70 on the cell surface, and this HSP70 activates NK cells (kurosawa, S. et al., Eur. J. lmmunol., 23: 1029, 1993). Cancer immunity can also be activated through the expression of. It has also been shown that the resistance to infection increases as the expression of HSP in host macrophages increases (Denagel, DC et al., Crit. Rev. lmmunol., 13:71, 1993). ), Immunostimulatory-biological defense ability enhancement effect can also be expected.

  Focusing on the function of intracellular protein quality control by HSP70, diseases in which abnormal proteins accumulate in the cells, such as Alzheimer's disease caused by β-amyloid deposition, Creutzfeld-Jakob disease in which abnormal prion protein is deposited, amyloidosis, It can be expected to be effective in preventing and treating degenerative diseases such as Wilson disease and Parkinson disease.

  It is not only expected to be resistant to physical stresses such as surgery and trauma, but also to suppress the onset and exacerbation of allergic diseases, stress ulcers and chronic inflammatory diseases caused by mental stress. I can expect. HSP70 improves the prognosis of multiple organ failure and shock due to sepsis (Hauser, GJ et al., Am. J. Physiol., 271: H2529, 1996) and adult respiratory distress syndrome It has also been reported (Villar, J. et al., Am. Rev. Respir. Dis., 147: 177, 1993), and its effect is expected as a therapeutic agent during these severe bioinvasions.

  Since HSP is an intracellular (in vivo) substance, it is unlikely that side effects will occur with its induction. In addition, no diseases caused by overexpression of HSP70 have been reported. In animal experiments, systemic heat shock, transient ischemia manipulation, HSP70 gene transfer, etc. are carried out, but it is difficult to apply to actual clinical practice. For this reason, it can be said that a device that does not harm tissues and cells and selectively induces HSP is a clinically superior treatment device.

  Since transcription factors are located at the bottom of the signal transduction system from the outside world, it is considered that side effects can be minimized by targeting them. NF-κB is one of transcription factors and is inactivated by binding to IκB, which is an inhibitory protein, in the cytoplasm. When cells undergo various stimuli, IκB is phosphorylated and subsequently ubiquitinated and degraded by the proteasome. The released NF-κB moves to the nucleus and specifically activates various genes. Examples of genes under the control of NF-κB include cytokines (TNF-α, β, IL-2, 6, 8, etc.) that play an important role in cells of the immune system. Since these genes are induced when cells are stimulated, it can be seen that NF-κB is deeply involved in the immune response. However, it is known that when the inflammatory response is excessive, various diseases are caused. For example, NF-κB is controlled because NF-κB is involved in various inflammatory diseases such as rheumatism, asthma and dermatitis, autoimmune diseases, viral diseases, arteriosclerosis, etc. The significance of is extremely great clinically (Anning Lin. Cancer Biology, 2003, Aggarwal BB et al. Indian J Exp Biol, 2004, Alok C. Bharti et al. Biochemical Pharmacology, 2002).

  However, the biological normalization mechanism based on the ubiquitin-proteasome system is capable of rapidly and reliably degrading unnecessary proteins because the amount of ubiquitin present in all cells is not sufficient when the biological tissue is normal (healthy). Had a problem that could not be eliminated. In the case of abnormalities (diseases) in living tissues, ubiquitin-bound proteins are decomposed and eliminated, so the intracellular ubiquitin is also reduced, and the cells of this abnormality (disease) are planned. Thus, there is a problem that abnormal cell death does not occur, abnormal cells such as tumors do not decrease, and abnormalities (diseases) cannot be improved.

  On the other hand, the living body normalization mechanism based on HSP by an electric thermotherapy device or the like does not sufficiently induce HSP expression and is not activated unless a very high temperature (for example, 42 ° C.) is heated to the living body for 1 hour or more. For this reason, there is a problem that rapid and accurate normalization of the living body cannot be performed, accompanied by damage to the living tissue due to heating. In addition, there was no method for normalizing a living body by affecting the amount of IκB that controls the activity of NF-κB and the phosphorylation state.

In order to solve these problems, the inventors have proposed a biological tissue normalization method capable of quickly and accurately normalizing a living body or a living tissue in Patent Document 16. The technique shown in Patent Document 16 intermittently applies a weak direct current to a living body or a living tissue at a predetermined interval, and further applies heat to change the normalization mechanism in the living body or the living tissue via a protein. It is a technology to activate. The proteins to be activated are mainly ubiquitinated proteins, heat shock proteins, IκB proteins and the like. It is also shown that the applied voltage (tester voltage value) is desirably set to 0.3 V, the frequency is set in the range of 50 Hz to 60 Hz, and the temperature of the heat is set to 38 degrees to 45 degrees. In Patent Document 16, evaluation in a human cultured cell system, evaluation in a mouse living body, and evaluation in a cultured cell system in which a mutant protein (ΔF508CFTR) is stably and highly expressed are performed. In any case, it has been shown by experiments that induction of HSP, promotion of ubiquitination, and increase in IκB and IκB-α phosphorylated forms are remarkable when a weak current and heat are used in combination. That is, it is clear that the treatment apparatus shown in Patent Document 16 has an extremely excellent HSP induction ability and is effective for various diseases by having an antitumor effect by inhibiting the proteasome. Moreover, in order to moderately suppress the phosphorylation of IκB by increasing the amount of IκB, it is expected to improve various pathologies caused as a result of an excessive immune response by NF-κB. Furthermore, from the aspect of safety, it can be expected to have extremely excellent clinical usefulness.
Samali, A.et ai., Cell Stress & Chaperones 3: 228,1998 Minowada, G. et al., J. Clin. Invest., 95: 2,1995 and references cited Kitagawa, K. et al., J. Cereb. Blood Flow Metab., 11: 449, 1991 Donnelly, TJet al., Circulation 85: 1048,1992 Maber, MSet al., J. Clin. Invest., 95: 1446, 1995 Polla BSet al., Proc. Natl. Acad Sci. USA, 93: 6458, 1996 Koenig, WJet al., Plast Recontsr. Surg., 90: 659,1992 Flohe, S. et al., Traspl. Lnt., 11: 89,1998 kurosawa, S.et al., Eur.J.lmmunol., 23: 1029,1993 Denagel, DCet al., Crit. Rev. lmmunol., 13:71, 1993 Hauser, GJet al., Am. J. Physiol., 271: H2529, 1996 Villar, J. et al., Am. Rev. Respir. Dis., 147: 177,1993 Anning Lin. Cancer Biology, 2003 Aggarwal BB et al. Indian J Exp Biol, 2004 Alok C. Bharti et al. Biochemical Pharmacology, 2002 JP 2006-263456 A

However, the value of the voltage value in the technique of Patent Document 16 indicates the value of the tester voltage, which is different from the actually input voltage value. In other words, since the voltage is applied so that the value of the tester is 0.01 V or more and 0.4 V or less which is considered to be actually effective,
The actual input voltage value is not clear.
In addition, when it takes time for the voltage to rise, there is a problem that a sufficient effect may not be obtained due to the defense function.

  The present invention has been made in order to solve the above-described problems. In the method of activating a normalization mechanism of a living body or a living tissue through a protein by applying a weak current and heat, the most effective voltage value and standing are provided. It is an object of the present invention to provide a biological tissue normalization method and a treatment apparatus that activates a protein with a raising time.

(1. Biological tissue normalization device)
The biological tissue normalization apparatus according to the present invention applies a weak direct current generated by intermittently applying a DC voltage at a predetermined interval to a living body or living tissue, and adds heat to the living body or living tissue. In the biological tissue normalization apparatus that activates a normalization mechanism in the living body or biological tissue via protein, the pulse waveform of the applied input voltage is a rectangular wave, and the rising peak value in one cycle of the pulse wave is The time shown is 0.05 milliseconds or more and 0.2 milliseconds or less, and the peak value is 3.0 V or more and 20.0 V or less.

  Thus, in the present invention, the pulse waveform of the input voltage to be applied is a rectangular wave, and the time indicating the rising peak value in one cycle of the pulse wave is 0.05 milliseconds or more and 0.2 milliseconds or less. Since the peak value is 3.0 V or more and 20.0 V or less, the input value of the voltage to be applied is clarified, and there is an effect that normalization of the living body or the living tissue can be performed effectively.

The time for showing the peak value of the rising edge in one cycle of the pulse wave is preferably 0.05 milliseconds to 0.2 milliseconds, more preferably 0.05 milliseconds to 0.15 milliseconds. Yes, more preferably 0.1 milliseconds.
The rising peak value in one cycle of the pulse wave is preferably 3.0 V or more and 20.0 V or less, more preferably 3.0 V or more and 16.8 V or less, and further preferably 12.0 V at the animal level. The cell level is 3.0V to 7.0V.

(2. Rise time)
The biological tissue normalization apparatus according to the present invention has a rise time of the pulse wave of 18 nanoseconds or more and 5000 nanoseconds or less.

As described above, in the present invention, since the rise time of the pulse wave is an extremely short time of 18 nanoseconds or more and 5000 nanoseconds or less, the stimulation to the living body or the living tissue becomes clear, and the function of the defense function is suppressed. There exists an effect that normalization of a living body or living tissue can be performed very effectively.
The rise time of the pulse wave is preferably 18 to 5000 nanoseconds, more preferably 18 to 1000 nanoseconds, and further preferably 18 nanoseconds.

(3. Relationship between peak value and peak time)
The biological tissue normalization apparatus according to the present invention is such that the time and peak value indicating the rising peak value in one cycle of the pulse wave are inversely proportional.
As described above, in the present invention, the time indicating the peak value of the rising edge in one cycle of the pulse wave and the peak value are in an inversely proportional relationship, so that the current stimulus and the thermal stimulus can be appropriately balanced, There exists an effect that normalization of a living body or a living tissue can be performed effectively.

(4. Heat shock protein and ubiquitin protein)
In the biological tissue normalization apparatus according to the present invention, the protein is a heat shock protein or a ubiquitinated protein.
As described above, in the present invention, since the protein is a heat shock protein or a ubiquitinated protein, it can be effective for various diseases and has an antitumor effect by inhibiting the proteasome.

(5. DC current flow interval)
In the biological tissue normalizing apparatus according to the present invention, the intermittent interval of the DC voltage is 30 pps to 100 pps.
As described above, in the present invention, since the intermittent interval of the DC voltage is 30 pps to 100 pps, normalization of the living body or the living tissue can be effectively performed by appropriately applying the current stimulus. There is an effect.

(6. Temperature of heat)
In the biological tissue normalizing apparatus according to the present invention, the thermal temperature is not lower than 38 ° C and not higher than 45 ° C.
Thus, in the present invention, since the heat is 38 ° C. or higher and 45 ° C. or lower, there is an effect that normalization of the living body or the living tissue can be effectively performed by appropriately applying the thermal stimulus.

(7. Treatment device)
The treatment apparatus according to the present invention is provided with a conductive layer made of a conductive sheet body in which a pair of pad elements adheres to the surface of different parts of a living body or a living tissue, and disposed on the back side of the conductive layer. An insulating layer made of an insulating sheet having characteristics and disposed on the back side of the insulating layer, a pair of electrodes are disposed at both ends, and a resistance is disposed between the pair of electrodes. A voltage control means for applying a voltage for generating a current between the pad elements to generate a weak DC current between the conductive layers of the pair of pads. Are controlled intermittently at predetermined intervals, and the input voltage for generating a current between each pair of electrodes in each heat generating layer of the pair of pad elements is controlled to generate the direct current. Input voltage is a rectangular pulse waveform The time for showing the peak value of the rising edge in one cycle of the pulse wave is 0.05 milliseconds or more and 0.2 milliseconds or less, and the peak value is 3.0 V or more and 20.0 V or less at the cell level. is there.

  Thus, in the present invention, the pulse waveform of the input voltage to be applied is a rectangular wave, and the time indicating the rising peak value in one cycle of the pulse wave is 0.05 milliseconds or more and 0.2 milliseconds or less. Since the peak value is 3.0 V or more and 20.0 V or less at the cell level, the input value of the voltage to be applied is clarified, and normalization of the living body or the living tissue can be effectively performed. Play.

A function generator may be used for voltage input. By doing so, the input voltage can be applied accurately, and the circuit can be simplified.
The time for showing the peak value of the rising edge in one cycle of the pulse wave is preferably 0.05 milliseconds to 0.2 milliseconds, more preferably 0.05 milliseconds to 0.15 milliseconds. Yes, more preferably 0.1 milliseconds.
Furthermore, the peak value of the rising edge in one cycle of the pulse wave is preferably 3.0 V or more and 20.0 V or less, more preferably 3.0 V or more and 16.8 V or less, and further preferably 12.0 V at the animal level. The cell level is 3.0V to 7.0V.

(8. Rise time)
The treatment apparatus according to the present invention is characterized in that the rise time of the pulse wave is 18 nanoseconds or more and 5000 nanoseconds or less.
As described above, in the present invention, since the rise time of the pulse wave is an extremely short time of 18 nanoseconds or more and 5000 nanoseconds or less, the stimulation to the living body or the living tissue becomes clear, and the function of the defense function is suppressed. There exists an effect that treatment can be performed very effectively.
The rise time of the pulse wave is preferably 18 to 5000 nanoseconds, more preferably 18 to 1000 nanoseconds, and further preferably 18 nanoseconds.

(9. Relationship between peak value and peak time)
The treatment device according to the present invention is characterized in that the time indicating the peak value of the rising edge in one cycle of the pulse wave and the peak value are in an inversely proportional relationship.
As described above, in the present invention, the time indicating the peak value of the rising edge in one cycle of the pulse wave and the peak value are in an inversely proportional relationship, so that the current stimulus and the thermal stimulus can be appropriately balanced, There exists an effect that a treatment can be performed effectively.

(10. Formation of resistance by carbon fiber)
In the treatment device according to the present invention, the heat generating layer of the pad element is a carbon fiber having a pair of electrodes each formed in a strip shape, and a resistance between the electrodes being oriented in a direction parallel to the strip electrode. It is formed.
Thus, in the present invention, the heating layer of the pad element is a carbon fiber in which a pair of electrodes are each formed in a strip shape, and the resistance between the electrodes is oriented in a direction parallel to the strip electrode. Since it is formed, the electrodes can be heated by an appropriate resistance value between the parallel carbon fibers without being short-circuited by the carbon fibers, and the treatment can be effectively performed.

(11. DC current application interval)
The treatment apparatus according to the present invention is characterized in that the direct current voltage supplied to each of the conductive layers is intermittently controlled in a cycle of 50 pps to 60 pps.
Thus, in the present invention, since the intermittent interval of the DC voltage is 50 pps to 60 pps, the treatment can be effectively performed by appropriately applying the current stimulation.

(12. Temperature of heat)
The treatment apparatus according to the present invention is characterized in that each heat generating layer of the pair of pad elements is heated to 38 ° C. or higher and 45 ° C. or lower.
Thus, in this invention, since heat is 38 degreeC or more and 45 degrees C or less, there exists an effect that treatment can be performed effectively by giving a thermal stimulus moderately.

(First embodiment of the present invention)
Hereinafter, a treatment apparatus using the biological tissue normalizing apparatus according to the first embodiment of the present invention will be described with reference to FIGS. 1 is an overall schematic configuration diagram of a treatment apparatus according to the present embodiment, FIG. 2 is a plan view of a pad element in the treatment apparatus described in FIG. 1, and FIG. 3 is a cross-sectional view taken along line AA of the pad element described in FIG. 4 and 4 are sectional views taken along line BB of the pad element shown in FIG.

  In each of the drawings, the treatment apparatus according to the present embodiment attaches a pair of pad elements 1 and 2 to different parts of the living body 100, and from a power source to generate current between the pair of pad elements 1 and 2. In the treatment apparatus for treating the living body 100 by controlling the input voltage supplied by the voltage control means 3, the conductive layers 11, 21 are made of a conductive sheet on which the pad elements 1 and 2 adhere to the surface of the living body 100. The insulating layers 12 and 22 are disposed on the back side of the conductive layers 11 and 21 and have a heat transfer characteristic and are made of an insulating sheet. The insulating layers 12 and 22 are disposed on the back side of the insulating layers 12 and 22. A pair of electrodes 13a, 13b, 23a, 23b are disposed at both ends, and heat generating layers 13, 23 are provided with resistors 13c, 23c between the pair of electrodes 13a, 13b, 23a, 23b, The heating layers 13 and 23 And covering layers 14 and 24 made of an insulating sheet that is disposed on the outermost surface and having a heat insulating property, and the voltage control means 3 includes the conductive layers 11 in the pair of pad elements 1 and 2. , 21 to control the supply of input voltage intermittently at predetermined intervals in order to generate a weak DC current between the pair of electrode elements 1 and 2 in the heat generating layers 13 and 23 of the pair of pad elements 1 and 2. In this configuration, the input voltage is supplied and controlled in order to generate a direct current between 13a and 13b and 23a and 23b.

  In the heat generating layers 13 and 23, a pair of electrodes 13a and 13b and 23a and 23b are each formed in a strip shape, and resistors 13c and 23c between the electrodes 13a and 13b and 23a and 23b are formed in a strip shape. , 23a and 23b, the carbon fiber 16 having an orientation in a direction parallel to the structure, and heated to a heating temperature of 42 ° C.

  The voltage control means 3 supplies the voltage only for 10 minutes or more and 30 minutes or less so that the conductive layers 11 and 21 are intermittently turned on at a period of 55 pps and controls the input voltage to be applied. It is.

  The input voltage applied to the living body or living tissue by the voltage control means 3 and the frequency of intermittent application will be described based on test results for human subjects. This test of human subjects is performed at 7.2 [V] to 26.4 [V], preferably 7.2 [V] to 16.8 [V], more preferably 12.0 [V] at the application site on both human feet. The voltage control means 3 controls the input voltage of V].

  In this human subject test, an applied voltage of 12.0 [V] is applied to both human feet in the subject, and the frequency is changed from 35 [pps] to 150 [pps], and the subject's senses (comfort feeling) -Uncomfortable feeling) was detected as follows.

  First of all, at 35pps, it was still uncomfortable, and it was found that feelings became worse after prolonged processing. It was found that at 45 pps, the feeling of numbness was strong, the patient felt uncomfortable (uncomfortable), and the treatment for a long time was impossible. It was found that 50 pps is an acceptable range of comfort. It turns out that it is very comfortable at 55 pps. It was found that 60 pps is an acceptable range of comfort.

  Further, at 65 pps, a little stimulation was not felt, and when the input voltage was increased to 14.4 [V], a stimulation similar to that at 55 pps could be felt, but it was found that it was not optimal. It was found that almost no irritation was felt at 70 pps. It was found that at 75 pps, almost no stimulation was felt, and when the input voltage was increased to 16.8 [V], a slight stimulation was felt. It was found that at 100 pps, no stimulation was felt at all, and when the input voltage was increased to 19.2 [V], a slight stimulation was felt. It was found that at 150 pps, no stimulation was felt and the input voltage was raised to 26.4 [V] but not.

  Moreover, the optimal frequency of the frequency which applies this direct-current voltage intermittently when input voltage was set to 9.6 [V], 12.0 [V], and 16.8 [V] was calculated | required by the test. When the frequency is changed at 35 <pps <50 when the input voltage is 9.6 [V], the feeling of numbness is felt strongly and unpleasant at 35 [pps] or less, and not felt at 50 [pps] or more. Further, when the frequency is changed at 45 <pps <60 when the input voltage is 12.0 [V], the feeling of numbness is felt strongly and unpleasant at 45 [pps] or less, and not felt at 75 [pps] or more. When the input voltage is 16.8 [V] and the frequency is changed at 65 <pps <75, the muscle contraction appears strong or unpleasant at 75 [pps] or less, and 75 [pps] or more. I don't feel.

  From the above test results, at the animal level, the input voltage is 12.0 [V] and the frequency 55 (± 1) [pps] is the optimum frequency, and the voltage control means 3 is in the range of 50 [pps] to 60 [pps]. It can be seen that it is desirable to control.

Moreover, the voltage control means 3 can control the input voltage as follows from the function of the electrical signal to the living body or the living tissue. That is, a voltage is forcibly applied to a non-excitable cell of a living body or a living tissue so that a weak current corresponding to a living body current is applied, and activation is performed through a protein in the body. In this way, only a non-excitable cell is supplied with a weak current corresponding to a bioelectric current, and an excitable cell such as a muscular cell is not supplied with an external current corresponding to a bioelectric current. Absent.
Therefore, the voltage control means 3 controls the input voltage to be applied so that the excitable cells pass a weak current at a current level that does not cause the muscle cells to contract uncomfortablely.

Hereinafter, the effect of inducing the protein expressed by the operation of the treatment apparatus according to the present embodiment will be specifically described based on the results of experiments conducted on human cultured cell systems.
(1. Items to be examined)
Evaluation in a human cultured cell line (human lung cancer cell A549 cells) was performed on the following two points i) and ii).
(1-1) Induction of HSP72 (1-2) Promotion of ubiquitination

(2. Implementation method)
The input voltage applied by a function generator (hereinafter referred to as FG) was set to 5.0 [V], the rise time was set to 18 nsec, and the following respective verifications were performed by changing the time indicating the peak value in various ways. . Moreover, it verified by setting the temperature of the heat to add to 42 degreeC.
(2-1 Induction of HSP72)
The measurement of HSP70 was performed by immunoblotting using a mouse anti-HSP70 monoclonal antibody, and the bound antibody was detected and measured by an enhanced chemiluminescence (ECL) Western blot detection kit (manufactured by Amersham). Calnexin (CNX) was detected as a loading control.
(2-2 Promotion of ubiquitination)
Ubiquitinated protein is measured by immunoblotting using mouse anti-ubiquitinated protein monoclonal antibody, and the bound antibody is detected and measured by Enhanced Chemiluminescence (ECL) Western Blot Detection Kit (Amersham). did.

(3. Implementation results)
(3-1 Induction of HSP72) (See FIG. 5)
FIG. 5 shows temperature + FG voltage (peak time 0.05 msec), temperature + FG voltage (peak time 0.1 msec), temperature + FG voltage (peak time 0.15 msec), temperature + FG voltage (peak time 0.2 msec), temperature + FG. The expression level of HSP72 in each case of voltage (peak time 1 msec) is shown.

From the graph shown in FIG. 5, it can be seen that the expression level of HSP72 is the highest when the peak time is 0.1 msec. In addition, when the peak times are 0.05 msec and 0.15 msec, the expression level of HSP72 is smaller than that when the expression level of HSP72 is 0.1 msec, but the expression level is increased. That is, it can be said that the peak time is preferably set to 0.05 msec to 0.2 msec, more preferably 0.05 msec to 0.15 msec, and still more preferably 0.1 msec.
The input voltage to be applied is set to 3.0 [V] to 7.0 [V], preferably 5.0 [V] at the cell level.
Furthermore, the voltage rise time is set to 18 nsec to 5000 nsec, preferably 18 nsec. By doing so, stimulation to living tissue and cells becomes clear, and normalization of living tissue can be performed extremely effectively by suppressing the function of the defense function

(3-2 Promotion of ubiquitination) (see FIG. 6)
HS indicates the presence or absence of heat, and + indicates that heat is applied and − indicates that heat is not applied. Moreover, the time which shows a peak value was 0.05 msec, 0.1 msec, 0.15 msec, 0.2 msec, and 1 msec, and the ubiquitinated protein in each case was detected by Western blotting. Again, it can be seen that ubiquitination is promoted over a peak time of 0.05 msec to 0.15 msec. In particular, the acceleration of ubiquitination can be confirmed most in the case of 0.1 msec.

  As described above, it is clear that the biological tissue normalization apparatus and the treatment apparatus using the biological tissue normalization apparatus according to the present invention have extremely excellent HSP induction ability and are effective for various diseases. In addition, the biological tissue normalization apparatus and the treatment apparatus using the biological tissue normalization apparatus according to the present invention can be expected to have extremely excellent clinical usefulness even from the viewpoint of high safety. Specific examples of the various diseases include cranial nerve disease, cardiovascular disease, digestive system disease, metabolic disease, autoimmune disease, degenerative disease, ischemic nerve cell injury, ischemia / reperfusion injury, cystic fiber Diseases, malignant tumors, infectious diseases, liver failure, renal failure, drug poisoning, heavy metal poisoning, radiation injury, ultraviolet ray injury, biological invasion, aging, etc. The cranial nerve disease includes stroke, sequelae of stroke, delayed neuronal cell death, Alzheimer's disease, Parkinson's disease, multiple sclerosis or Creutzfeld-Jakob disease.

  Moreover, the treatment apparatus using the biological tissue normalization apparatus and the biological tissue normalization apparatus according to the present invention activates a normalization mechanism in a living body or a biological tissue via an eviquitinated protein, About 80% of proteins are degraded by the proteasome after being ubiquitinated. However, if the function of the proteasome is inhibited, ubiquitinated proteins that are not broken down in the cell increase, and the cell chooses a planned path for cell death. Using this principle, proteasome inhibitors are currently attracting attention as anticancer agents (Julian A. Cancer Cell, 2003, Angelika M. B et al. European Journal of Cancer, 2004).

  Proteasome inhibitors tend to act during the growth phase when protein synthesis and degradation are active. Compared with normal cells, protein dysregulation related to cell growth occurs in tumor cells, and thus the cell growth rate is very fast. Therefore, tumor tissues are susceptible to proteasome inhibitors that act on proliferating cells. Since the biological tissue normalization apparatus and the treatment apparatus using the biological tissue normalization apparatus according to the present invention promote ubiquitination, intracellular ubiquitinated proteins greatly increase. For this reason, the proteasome is saturated, and the state is the same as when the proteasome function is inhibited.

  That is, the treatment apparatus using the biological tissue normalization apparatus and the biological tissue normalization apparatus according to the present invention also has an antitumor effect by inhibiting the proteasome. Moreover, the effect can be expected to be exhibited specifically for tumor cells based on these principles. Examples of diseases that can be improved by the normalization of the ubiquitin / proteasome system via ubiquitinated proteins include neurodegenerative diseases such as Parkinson's disease, Alzheimer's disease, amyotrophic lateral sclerosis (ALS), and myoclony. Epilepsy and the like), cancer diseases (for example, familial breast cancer, ovarian cancer, etc.), xeroderma pigmentosum and the like.

  Although the present invention has been described in the above embodiment with reference to animal tissue cells of human cultured cell lines, it can also be applied to plant tissue cells, and the same actions and effects can be achieved. It is done.

  Further, the input voltage from the function generator at the cell level is set to 3.0 [V] to 7.0 [V], preferably 5.0 [V], but 7.2 [V] at the animal level. ] By setting FG with 16.8 [V] or less, preferably 12.0 [V] as the input voltage, the same effect as described above can be obtained.

1 is an overall schematic configuration diagram of a treatment apparatus according to a first embodiment. It is a top view of the pad element in the treatment apparatus of FIG. It is the sectional view on the AA line of the pad element of FIG. FIG. 5 is a cross-sectional view of the pad element shown in FIG. 3 taken along the line BB. It is an evaluation result of induction of HSP72. It is an evaluation result of promotion of ubiquitination.

Explanation of symbols

DESCRIPTION OF SYMBOLS 1, 2 Pad element 3 Voltage control means 11, 21 Conductive layer 12, 22 Insulating layer 13, 23 Heat generating layer 13a * 13b, 23a * 23b Electrode 16 Carbon fiber 14, 24 Cover layer 100 Living body 200 Power supply

Claims (12)

A weak direct current generated by intermittently applying a DC voltage at predetermined intervals is applied to a living body or a living tissue, and heat is applied to the living body or the living tissue, and the normalization mechanism in the living body or the living tissue is changed to a protein. In the body tissue normalization device to be activated through
The pulse waveform of the applied input voltage is a rectangular wave, and the time indicating the rising peak value in one cycle of the pulse wave is 0.05 milliseconds to 0.2 milliseconds, and the peak value is 3. A biological tissue normalizing device, characterized in that it is 0 V or more and 20.0 V or less. In the biological tissue normalization apparatus according to claim 1,
The apparatus for normalizing a biological tissue, wherein the rise time of the pulse wave is 18 nanoseconds or more and 5000 nanoseconds or less. In the biological tissue normalization apparatus according to claim 1 or 2,
A biological tissue normalizing apparatus, wherein a time indicating a peak value of a rising edge in one cycle of the pulse wave and a peak value are in an inversely proportional relationship. The biological tissue normalization device according to any one of claims 1 to 3,
The biological tissue normalization apparatus, wherein the protein is a heat shock protein or a ubiquitinated protein. The biological tissue normalization device according to any one of claims 1 to 4,
An apparatus for normalizing a living tissue, wherein an intermittent interval of the DC voltage is 30 pps to 100 pps. The biological tissue normalization device according to any one of claims 1 to 5,
The living tissue normalizing apparatus, wherein the thermal temperature is 38 ° C or higher and 45 ° C or lower. A treatment device using the biological tissue normalization device according to claim 1,
A pair of pad elements are disposed on the back side of the conductive layer having a conductive sheet that adheres to the surface of a different part of a living body or living tissue, and has a heat transfer characteristic and an insulating property. An insulating layer composed of a sheet body, and a heat generating layer disposed on the back side of the insulating layer, a pair of electrodes disposed on both ends, and a resistance disposed between the pair of electrodes,
The voltage control means for applying a voltage for generating a current between the pad elements intermittently changes the input voltage at a predetermined interval so as to generate a weak DC current between the conductive layers of the pair of pads. Supply control, and supply control of an input voltage for generating a current between each pair of electrodes in each heat generating layer of the pair of pad elements,
The input voltage for generating the direct current is a pulse waveform of a rectangular wave, and the time showing the rising peak value in one cycle of the pulse wave is 0.05 milliseconds or more and 0.2 milliseconds or less, The treatment apparatus, wherein the peak value is 3.0 V or more and 20.0 V or less. The treatment device according to claim 7,
The therapeutic apparatus characterized in that the rising time of the pulse wave is 18 nanoseconds or more and 5000 nanoseconds or less. The treatment device according to claim 7 or 8,
A treatment apparatus characterized in that the time indicating the peak value of the rising edge in one cycle of the pulse wave and the peak value are inversely related. The treatment device according to any one of claims 7 to 9,
The heat generating layer of the pad element is characterized in that a pair of electrodes are each formed in a strip shape, and a resistance between the electrodes is formed of carbon fibers having an orientation in a direction parallel to the strip-shaped electrode. Therapeutic device. The treatment device according to any one of claims 7 to 10,
A treatment apparatus, wherein a direct current voltage supplied to each conductive layer is intermittently controlled at a period of 50 pps to 60 pps. The treatment device according to any one of claims 7 to 11,
The therapeutic apparatus, wherein each heat generating layer of the pair of pad elements is heated to 38 ° C. or higher and 45 ° C. or lower.