Method and device for inducing pathological cells and tissues apoptosis

FIELD: medicine; medical engineering.

SUBSTANCE: method involves acting upon pathological cells and tissues with constant and/or low frequency alternating electromagnetic field of 1 to 100 mTesla units large intensity and frequency of 1 to 1000 Hz. The device has means for producing constant and/or low frequency alternating electromagnetic fields permeating working space.

EFFECT: enhanced effectiveness in treating pathological tissues without harmfully influencing healthy cells.

19 cl, 5 dwg, 5 tbl

 

Scope.

The present invention relates to a device for influencing the life processes of abnormal cells.

In addition, the present invention relates to using this device microbiological way of influencing the life processes of abnormal cells, in particular cells, cancer and other diseases caused by changes in the mechanism of survival of the cells.

In particular, the electromagnetic constant field (C-field) and low-frequency electromagnetic field (ENC-field)generated by this device.

Magnetic constant field and low-frequency electromagnetic fields referred to herein as “C-field” and “ENC-field”, respectively. In addition, any combination of different types of sequences-fields, an fields, e.g. fields, followed by EN-field, Inc-field, followed by the C-field C-field, acting jointly with EN-fields and action fields, an fields individually hereinafter referred to as SAN-fields.

Prior art

It is known that extracellular fields and currents induced by electromagnetic fields (ENC-fields)whose frequency ranges from 1 Hz to 300 Hz, and possibly as high as 1000 Hz, cause in a specific cell membrane, electrochemic the sky changes which are important for the processes of transduction and amplification of the primary biological signal.

Data biochemically due to changes lead to the formation of cytoplasmic carriers of the second order and internal effectors, such as free CA++and protein phosphorylase (kinase), which, in turn, initiate some changes in the biosynthesis of macromolecules and lead to differentiation in the growth and functional properties of cells [1M. Blank, 1993].

Further, it was established the probability that and ENC-field effects on DNA synthesis, DNA integrity, transcription (the synthesis of complementary RNA on the matrix DNA or RNA - approx. interpreter and translation (protein biosynthesis on the information matrix RNA - approx. translator) [2Liboff 1984,3Tofani 1995,4Goodman 1991,5Philips 1992].

Possible physical mechanism responsible for some of the experimental findings, is a direct influence on the ion (i.e. the CA++) or by coupling the ligand on the cell membrane [6Liboff 1985,7Chiabrera 1985,8Lednev 1991,9Blanchard 1994].

The influence of variations in the metabolism of CA++can lead to cellular apoptosis (programmed death of cells) [10Preston,11Trump 1997].

Another mechanism of interaction is associated with the possibility of effect for the I on the kinetics of the corresponding cell signaling pathways that cells (including calcium metabolism), due to the direct impact of the field on the electron-spin motion of atoms and molecules with unpaired electrons. Such exposure can affect the recombination coefficient of the spin of the corresponding pair of free radicals and, consequently, redox signaling [12Grundler 1992,13Polk 1992,14Walleczek and Budingher 1992,15Adey 1993].

In particular, two-, three-energy level of the spin transition in a free radical is critical to increase the recombination coefficient of the spin of the corresponding pair of free radicals.

The probability for low-energy, non-thermal With and EN-field (magnetic induction of up to 30 MT) to influence in vitro on the kinetics and efficiency of the reactions of the pairs of radicals known from magnetokhimiya [16Steiner 1989].

Naturally occurring free radicals have an unpaired electron in the oxygen - or nitrogen-containing base, for example such as superoxide anion, hydroxyl radical and nitric oxide. Data reactive oxygen species (RVC) and reactive nitrogen species (ROS) can be targeted to proteins, providing obvious from the point of view of the mechanics of the explanation for the existence of phenomena signaling by free radicals. These phenomena may affect growth factors, transport of ions (i.e. channels of CA++), tra is scriptio (synthesis of complementary RNA on the matrix DNA or RNA approx. translator), apoptosis [17Lander 1997].

Apoptosis is a morphologically distinct form of programmed death of cells associated with the life processes of cells and plays an important role during development, homeostasis and in the case of many diseases, including cancer, acquired immunodeficiency syndrome, and neurodegenerative disorders and other diseases, similar to those which are characterized by variable processes of survival cells. Apoptosis occurs through activation of internal self-destruction of cells. The underlying genetic mechanism of apoptosis is present in almost all cells of the mammary gland during the whole time, but the rise of this self-destruction is regulated by many different signals coming from both intracellular and from the external environment.

Among all the genes involved in the regulation of apoptosis, the greatest attention should be paid to the gene p53. This gene, which encodes a transcription factor and is common in various types of human cancer, is the mediator of the cellular response to some disturbance. Protein p53 can either temporarily halt cell division so that the cell can recover the modified DNA, or may cause apoptotic destruction of the cell.

Published kovanye data confirm the fact, the gene p53 is present during apoptosis, thanks to the three stages of the process: 1) transcriptional induction of genes associated with the processes of oxidation-reduction; 2) the formation of reactive oxygen isotope and 3) oxidative decomposition of the components of the mitochondria, and this culminates in cell death [18Polyak 1997].

In addition, anti-oxidation agents are combined with medications in the treatment of hypoxic tumor cells19[Walch, 1988] and to influence the growth factor vascular20[Amirkhosravi, 1998].

Published data also confirm the fact that the abnormal cells compared to healthy cells otherwise respond to the impact of an fields. According to21Cadossi [1992] the lymphocytes of healthy individuals compared with lymphocytes of patients with down syndrome, AIDS patients, patients with chronic limfozitoza leukemia, otherwise respond to the impact of EN-fields (previously when exposed to mitogen).

Also is recognized by the fact that ANC-field influence the influx of CA++through the membrane when exposed to cells affected by leukemia, but not in the lymphocytes of healthy individuals [22Waleczek, 1996].

The life processes of the modified cells are electrical failures and with different electrical behavior of the cells. In particular, bystrorazvivajushchiesja and transformed cells, compared with healthy cells are depolarized the cell membrane [23Bingelli, 1986;24Marino 1994]. It was also shown that epithelial cells lose their transepithelial potential during oncogenesis [25Davies 1987;26Goller 1986;27Capko, 1996]. Such different electrical behavior of the cells of the tumors in comparison with healthy cells is the basis for a new diagnostic modality for cancer [28Cuzick 1998]. In addition, the concentration of free radicals in transformed cells and tissues is higher than in non transformed cells and tissues [29Szatrowski 1991;30Shulyakovskaya 1993;31Iwagaki 1995].

In the case of chemotherapy, all efforts are directed at ensuring tasks induction of apoptosis in vivo instead of killing cells by targeted therapies for signal transduction (NTSC) cancer [32Levin, 1998].

Signal transduction is a functional term that refers to the translation of genetic information in the signaling cascades that allow the cell to, for example, interpret and respond to external stimulation and/or duplicate itself. Recent studies show that a change in the process of survival of cells leads to the pathogenesis of some human diseases, including cancer, viral infections, autoimmune diseases, neurodegenerative the Sam the STS and AIDS. Treatment aimed at special change in the threshold of apoptosis-related mechanisms of the process of survival of the cells, can potentially alter the natural development of some of these diseases [33Thompson, 1995].

Electric, electromagnetic and magnetic fields with high magnetic induction is used to destroy abnormal cells.

In U.S. patent No. 466589834describes the device, where the animals who had malignant cells were subjected to treatment by pulsed magnetic fields with high magnetic induction in order to neutralize/destroy malignant cells selective way. Described in the patent, the device forms a thermal magnetic field with a magnetic induction of 1 Tesla 10 Tesla and reverse polarity within 5-1000 kHz. In a preferred embodiment, the magnetic induction of the magnetic field is from 1 to 50 Tesla and, in particular, in the examples at the level of 5 Tesla and 8 kHz to 18 Tesla and 250 kHz.

In anti-cancer therapy in vitro were used different EN-field, thermal field, continuous fields and fields with pulse modulation of [35Narita, 1997;36Raylman, 1996].

In these cases, the used fields with very high magnetic induction is much higher than allowed for a person in accordance with the standards without the provisions, that can lead to overheating and destruction of healthy tissues and cells.

Electromagnetic inch field with low magnetic induction was also used to stop mitosis malignant cells, as, for example, in German patent No. 4122380 A1 and U.S. patent No. 5156587. However, these documents describe the use of sinusoidal fields only at a fixed useful frequency and at a fixed magnetic induction, with the possible fluctuations of the energy levels inside the cellular tissue only in a limited range.

Brief description of the invention

The present invention is a method of influence on cellular processes of survival (i.e. stimulation of apoptosis) living pathological (i.e. cancerous) cells by using magnetic fields without harmful effect on healthy cells.

The present invention is also a device for the implementation of such influence on the process of survival of abnormal cells.

The principal and other tasks in accordance with the present invention are solved by the intervention activity of pathogenic cells by affecting the living of pathological cells (i.e. cancer cells and cells affected by other diseases, caused by the change of the mechanism of products is eljnosti cells) non-thermal magnetic C -, an-fields and thus, stimulation of selective apoptosis of cells.

For the purposes of the present invention under SANC-fields should understand the different sequence With courses and/or EN-fields, i.e. fields, followed by EN-field, Inc-field, followed by the C-field, and ANC-field acting jointly, as well as With fields, ENC-field acting separately.

The concept, which is based on a method in accordance with the present invention, is that SAN-field effect on the cell signals, confirming the development of a pathological process within the abnormal cells - i.e., the signals on redox potential of free radicals, and thus restore the processes of survival of cells, i.e. directly or indirectly stimulate apoptosis by modifying gene expression of p53.

This method presumably disconnects in free radicals with oxygen basis and can also be used as antioxidant agent. The application of this method can be considered in combination with the drugs for the treatment of hypoxic cells in tumors and to impact factor vascular growth.

The reason that SAN-field selectively stimulate apoptosis in pathological cells (i.e. cancer cells), may be treason is Mr. electric behavior of abnormal cells, compared with healthy cells.

For these reasons, SAN fields directly or indirectly can cause signal programmed cell death (apoptosis)in vitro and in vivo, without causing adverse effect.

In the hypothesis that recombination of free radicals is at the basis of the expected biological effect on pathological cells (i.e. anti-tumor activity), you must take into account the transition between the singlet-triplet unpaired electrons in free radicals with oxygen basis. In particular, this transition, which depends on the applied magnetic field, is critical for increasing the recombination coefficient spincorrelation pair of free radicals. However, the reaction centres related to the expected antitumor effects, is unknown, and therefore the duration of the spin state and distribution of energy between the singlet and triplet States cannot be accurately determined based on the spin Hamiltonian [37Haberkorn 1979,38Lersch 1983].

To cover this problem, in accordance with the present invention it is possible to use sequences With magnetic fields of different magnetic induction, amplitude modulated, and with the imposition of the magnetic ANC fields. The use of modulated fields meets the need the value of optimal conditions (conditions) to convert the singlet-triplet spin state, necessary for the recombination of free radicals [13Polk 1992].

For these reasons, field, ENC-field or SANC-fields are more likely to cause the expected biological effect, if they are modulated according to a given function of the magnetic induction and/or frequency from time to time, because in this case more likely to cause the above-mentioned transition.

Different sequences With courses and/or sequences of ANC fields were mainly established for time intervals T1T2,..., Tnwhere magnetic induction fields IC, IENand the ratio of Ic/IENset constant values of IC1, IC2,..., ICn; IEN, IEN;..., Iencp; IC1/IEN, IC2/IEN,...ISP/Iencprespectively.

For the same reasons modulated nonthermal of SAN-field can potentially be used for treatment of cells affected, for example, from diseases such as viral infection, AIDS, autoimmune diseases and so on, where the change process of the survival of cells additionally affects pathogenesis.

According to another aspect of the present invention, the device for selective intervention in the processes of survival of abnormal cells in vitro and in vivo is characterized by the fact that it sod is RIT means for generating a constant magnetic field (C-field), chilly working environment, and means for generating low-frequency electromagnetic fields in the work environment either separately or in addition to the fields.

Means providing a modulated With fields, associated with the means for generating the C-field and magnetic induction in the range from 1 to 100 MT, preferably from 1 to 30 MT.

Also provides a means to modulate an-fields, separate or connected with courses, with a frequency of 1 to 1000 Hz and magnetic induction from 1 to 30 MT. Preferably using an-fields with frequencies from 10 to 100 Hz.

In the private variant embodiment of the invention the means for modulating With fields include software tools that alternatively or in combination:

- set magnetic induction, following the many defined step-by-step interval of values of Ic1, IC2,..., Icnfor the respective time intervals t1T2,..., Tn;

- set the amplitude of the magnetic induction, following the many defined step-by-step interval of values of IEN, IEN,..., Iencpfor the respective time intervals T1T2,..., Tn;

- set frequency, following the many defined step-by-step interval of values of f1f2,..., fnfor the respective time sub-the Cove T 1T2,..., Tn;

- set the value of an according to the set specified with step-by-step interval of values of IC1/IEN, IC2/IEN,..., ICn/Iencpfor the respective time intervals t1T2,..., Tn.

Preferably, the software was installed With the-fields, an-fields in accordance with the total magnetic induction between 1 and 30 MT, respectively, the ratio of ANC fields in the range of from 0.1 to 10, and in some private versions - with the total magnetic induction from 1 to 10 MT, respectively, the ratio of ANC fields in the range of from 0.5 to 5.

Time intervals preferably be set from 1 to 40 minutes.

At least one part of the working environment is limited by the walls, permeable to the fields, an fields. At least one portion of the workspace is also preferably adjacent, respectively, to the first and second inductor, and means for modulating applying DC and AC current, respectively.

Brief description of drawings

Several variants of realization of the device according to the invention shown in the drawings are given as examples, but do not limit the possible options of device:

Figure 1 schematically shows a first variant of the device according to this image is the shadow;

Figure 2-4 shows the block diagram of the second, third and fourth variants of realization of the device according to the invention, respectively;

On Figa in the form of a diagram showing the function of the magnetic induction field from time programmable device in accordance with the claimed invention;

On Figv in the form of a diagram showing a function of time of the magnetic induction field for fields, EN-fields, with the change ratio in respect of each subsequent field;

On Figs in the form of a diagram showing the function of the magnetic induction field and frequency from time to time.

Description of the preferred embodiments of the device

In the drawing Figure 1 the working environment specified under number 1 and the wall 2. The first and second inductors are listed under numbers 3 and 4, respectively.

Means of modulation is shown schematically squares 5 and 6, respectively, and connected to the sources of AC and DC.

Another variant of the device that is used to influence the process of survival of abnormal cells in vitro and in vivo, as shown in the drawing, Figure 2, has two coils 23 and 24 are located on the same axis in relation to each other on opposite sides of the working environment 21. Variable transformers 25 and 26 connected to the electricity grid 27 AC is astotal 50 Hz. Plug diode bridge 28 is necessary to change the supply of alternating current to the coils. Transformer DC 29A, the rectifier 29b, and the timer 29s are attached to the two plates 29 so that it becomes possible to create in a production environment 21 DC electric field of 20 kV/m (or field of low frequency, changing to 1000 Hz, preferably about 6 kV/m, in the preferred intervals in accordance with the experimental conditions.

Figure 3 shows the following variant of the device for influencing the survival of abnormal cells in vitro with the modulator 35 SAN-fields (1-100 Hz) and two windings 33 and 34 located on one axis relative to each other on opposite sides of the working environment 31. The amplifier 36 is used between the modulator 35 and the coils 33 and 34, which receives the same current that creates a magnetic - or ENC - field in a production environment, 31.

Another variant of the device according to the invention (Figure 4) can be used to influence the survival of abnormal cells in vitro and in vivo and has two Helmholtz coils 43 and 44, which are located on the same axis on different sides of the operating environment 41. The amplifier 46 is used between the modulator 45 and the coils 43 and 44 through a shunt element 47, which is also connected to a personal computer 49.

Each of the devices can be used is to create a modulated nonthermal of SAN fields to influence the survival of abnormal cells.

The drawings from Figa to Figs shows an example of the programming of the device, when the modulation of the magnetic induction, the frequency and the ratio of magnetic induction between the With - and ANC-fields.

On Figa shows a variant in which the magnetic induction can vary with time. On this drawing I1, I2, I3, Inis the magnetic induction or the field strength (MT) for off-field or ANC-field individually, or the total magnetic induction of IC+IEN.

On FIGU when and EN-fields act together, it is possible to modulate not only the magnetic induction or the amplitude of the magnetic induction, but also their ratio - the ratio of IC/IEN. For example, different ratios of 1; 1,5; 2; etc. can be used for time intervals T1T2T3etc.

You can also modulate the frequency, as shown in Figs. The frequency can also be modulated in two or more time intervals T1T2using the same magnetic induction of I1-2.

Relying on basic examples Figa-5S, you can create a sequence modulated With, ENC -, - + ENC-fields that can also be cyclically repeated.

The method in accordance with the present invention is hereinafter described in more detail with specific examples.

Example 1

p> This example was studied in vitro the possibility of stimulation of apoptosis using magnetic SEN-field as a function of the magnetic induction and the frequency of the field.

For the experiment, was used cell line adenocarcinoma of human colon (WiDr)were grown in monolayers layer continuous culture on a solid nutrient medium in flasks KZT25. For each irradiation effects were used 6 of these flasks, each of which contained about 10 million cells, 3 of which were exposed to, and 3 were not exposed.

During irradiation, the flask was placed between the two inductors connected to the DC and AC currents with frequencies up to 100 Hz. Temperature was continuously monitored and maintained at 37±0,2°C.

For each experiment, the duration of exposure was 20 minutes and SANC-field was maintained unchanged. After three hours, cells were processed by May-Grunwald-Giemsa. Apoptosis was evaluated by counting the number of damaged kernels per 10 high power fields (VMP) using an optical microscope.

The number of induced apoptosis was evaluated by the ratio between the amount of damaged cells, found in the group exposed to, and the number of destroyed cells detected in the other group, Tav group, not exposed to magnetic fields in accordance with this invention.

Table 1 shows the results obtained under different conditions of exposure.

TABLE 1
Exposure conditionsThe structure of SANC-fieldFrequency (Hz)Magnetic induction field (effective value+Inc) MTLThe degree of apoptosis
AndWith (constant)-(0,5+0)1
In-(1+0)1
-(2+0)1,2
D-(3+0)2
E-(4+0)2,3
F-(10+0)2,2
G-(20+0)2,2
N-(30+0)2,3
IEN16(0+3)2,2
LEN33(0+3)2,2
MEN50(0+3)2,1
NEN50(0+7)2,1
0EN66(0+3)2,2
PEN83(0+3)2,3
QEN100(0+3)2,1
R+Inc50(4+3)2,1
S+Inc5050% of the time(3+1)

50% of the time (4,5+1,5)
2,2

All results were statistically extremely important (from the point of view of the test t Student). From Table 1 it is evident that the effect of apoptosis occurs when the value of an induction of about 1 MT and doubles from 3 MT.

Another important discovery was that apoptosis does not depend on the frequency of SEN fields. In other words, throughout the duration of the mechanism for biological outcome (apoptosis), ENC-field behaves as essentially a constant field. This means that among the two hypothetical mechanisms - mechanism involving free radicals (appearing in the time interval from the nano - to microseconds) and a mechanism that is similar to the ion resonance, jus the th role plays a mechanism involving free radicals [ 39Scaiano, 1994;40Engstrom, 1997].

Example 2

In this experiment, the electoral action of magnetic SEN fields was confirmed by impact against the three cell lines. Two lines were malignant: cell adenocarcinoma of human colon (WiDr) and cancer cells human breast (MCF-7). As healthy cells were selected cell lung fibroblast person (MRC-5).

In the same way as in example 1, each cell line was grown in monolayers layer continuous culture on a solid nutrient medium in flasks KZT25. The Protocol of the experiment was the same as in example 1. Six flasks of each cell line (3 - subject and three are not exposed to) was irradiated for 20 minutes. Apoptosis was evaluated after three hours. Exposure conditions were as in column “R” in Table 1.

The results are shown in Table 2.

TABLE 2.
Cell lineThe degree of apoptosis
WiDr2,1
MCF-71,4
MRC-51

As shown in Table 2, only the cancer cells showed a more significant from a statistical point of view the increase in apoptosis, whereas healthy cells such are not shown. It was also expected that there will be floor is Chen a different percentage of apoptosis between the two lines of cancer cells in two different time periods of reproduction. The fact that the cells of adenocarcinoma of human colon (WiDr) multiply faster cancer cells human breast (MCF-7). The results were evaluated on the basis of the test t of Student.

Example 3

In this example, to assess the impact of magnetic SEN fields on the suppression of tumor growth were derived mouse strain nude (nu/nu) with subcutaneous tumor weight.

Each mouse was injected subcutaneously with 10 million cells adenocarcinoma of human colon (WiDr). Were successfully conducted two experiments.

In the first experiment, 36 females were randomly divided into four groups, each consisted of 6-exposed mice and 3 are not affected, a total of 24 animals were exposed to 4 different magnetic SEN courses and 12 were not subjected to this influence.

Applied DC electric field of up to 6 kV/m for use ultimately benefits of different electrical behavior of the tumor tissue and healthy tissue [41Thornton, 1984;42Barsamian, 1987].

In the second experiment, 24 females were randomly distributed in two experimental groups of 12 mice exposed to SANC-field conditions, showed the best results among the four exposure conditions used in the previous experiment (impact 4), the 12 mice not affected.

All mouse participated in both experiments were divided into the experimental group after the tumor weight of each animal was palpable.

Animals were exposed for 70 minutes, once a day, for 5 days per week, for 4 weeks. During exposure, each mouse was placed in a box made of plexiglass, which is placed between the two inductors connected to the circuit of AC and DC to 100 Hz, respectively.

Mice nu/nu contained in special Bezmaternykh conditions, and they were fed according to the diet ad libitum” (no desire). All the tests were conducted according to the Protocol issued by the National Institute of health and National cancer Institute of the United States.

The weight of tumors were measured twice a week, and their volume was calculated in mm3according to the formula: {(larger diameter)×(smaller diameter squared)/2.

After 4 weeks the animals were euthanized, and they had done the autopsy. The tumor weight was removed, weighed and measured. Parts of the tumor were used for various analyses, namely:

- immunocytochemical: antigen Ki-67 index reproduction, antigen p-53 expression of the gene p-53;

- histopathology: staining hematoxylin-easingly estimates of the number of mitosis;

- ultrastructural: electron microscopy;

- hybridization of nucleic acids: a Tunneling method for assessment of apoptosis.

In addition, histological study to evaluate the toxicity of the treatment of each animal were extracted following organs: brain, heart, kidneys, liver, lungs, auxiliary and inguinal lymph nodes, mediastinal lymph nodes, ovaries, skin, spleen, bone marrow, subcutaneous tissue (part of the implanted lines of tumor cells), and were also made blood tests.

The results obtained are shown in Table 3 for the first experiment, and in Table 4 for the second experiment.

TABLE 3
Exposure conditions1234Not affected
The time of exposure (min)70707070-
Time-averaged magnetic induction field (effective value for the constant + ENC-field) in MT3346-
The variation of the magnetic induction in MT (min-max)-field; [min-max] ENC-field(4-6) [2-2](1,5-4) [1-1](2-5) [1,5-35] (2-5) [1,5-3,5]-
The time constant of the field (min-max) in minutes(5-15)(5-20)(5-15)(5-20)-
% of the time the joint action of the constant and ENC-field0%50%50%100%-
The ratio S-/EN-field (min-max)-(0.5 to 5)(0.5 to 5)(0.5 to 5)-
% time one constant field50%50%50%0%-
The number of mice666612
The amount of removed tumor mass (mm3)1323±3041450±288920±540650±2051492±559
The weight of the removed tumor mass (g)1,54±0,221,6±0,390,98±0,560,96±0,251,6±0,5
The number of apoptotic cells per 10 VMP (high power fields)98±23115±20129±25129±2640±17
The expression of p53 at 10 VMP35,1±0,1143,8±0,1638,2±0,06 28,7±0,1473,2±0,14

TABLE 4
Exposure conditions4 (see table 3)Not affected
The number of mice1212
The amount of removed tumor mass1139±509 cm31914±793 cm3
The weight of the removed tumor massof 1.4±0.7 gof 2.1±0.6 g
Apoptosis (assessed only in 50% of mice)72,5±9,337,0±7,4
P5335,6±6,778,1±16,7
The index reproduction0,34±0,080,45±0,07
Mitosis24,1±10,947,7±10,1

The data presented in tables 3 and 4 show that SAN-fields have the ability to stop the growth of tumors in a living organism (in vivo). This effect, discovered in the course of both experiments were very important from a statistical point of view (in the first experiment, mainly due to the exposure conditions under number 4) by Dunnet tests and Student t, respectively.

Histological examination of the 12 organs of each animal from each group was not detected no differences between podolskis and non-exposed mice. No difference was also not detected in the blood. These data show that toxic effects during therapy with the use of SAN-fields is missing.

Ultrastructural analysis by electron microscope showed that the tumor cells of animals subjected to impact occurred cellular changes: the presence of apoptotic bodies and condensed chromatin near the membrane of the cell nucleus indicates the presence of apoptotic phenomena.

In addition, a consistent result is confirmed by morphological modifications, increasing the number and size of mitochondria, and the number of nucleoli, the presence of many vacuoles within the cytoplasm. Not related to the tumor cells (i.e. epithelial cells and stromal cells) showed that there is no difference between animals exposed and non-exposed, which confirms the absence of toxic effects, as identified in 12 healthy bodies examined animals.

The increase in apoptosis and decrease in gene expression of p53 found in the affected tissues tumors in animals (see tables 3 and 4), have important statistical significance (Student t test).

The results presented in Tables 3 and 4, correspond to those obtained in vitro and displayed in Tables 1 and 2.

The effect proizvodityelnyye SAN-fields on gene expression of p53, enhances apoptosis that leads to the conclusion that full compliance with the hypothetical biophysical mechanism (i.e. recombination of free radicals), determining the antitumor effect of SAN-fields, due to the formation of active oxygen isotopes and decay of mitochondrial components.

Example 4

In this experiment mice nu/nu, which were first introduced under the skin 10 million cells adenocarcinoma of human colon (WrDr), were exposed to in order to study the survival of animals.

After injection of the cells were randomly formed two groups of mice, which consisted, respectively, of the 16 affected animals and 17 is not affected. Mice of the first group were exposed for 70 minutes a day, 5 days a week and throughout their lives, starting 24 hours after injection of tumor cells.

Exposure conditions were the same as in the experiment whose results are shown in Table 4.

As in the previous example, the mouse was kept in a special Bezmaternykh conditions, and fed them according to the diet “ad libitum”. All the tests were conducted according to the Protocol issued by the National Institute of health and National cancer Institute of the United States.

The efficacy of this therapy to combat the tumor is and was estimated by the formula of the National Institute of cancer - as the ratio of exposed and non-exposed animals, with an average life expectancy. This averaged value was calculated as the sum of the survival time for each experimental group divided by the number of animals. Efficiency is achieved, if the formula of the national cancer Institute of the USA gives a result equal to or greater than the value of 1.25.

Table 5 shows the data for each experimental group by the number of live animals at different times (days) from the beginning of the experiment.

When using formulas of the National cancer Institute of the USA to calculate the index based on the results in Table 5 produces the number of 1.31 - greater than 1.25. After 194 days 6 mice exposed to, were still alive, and all non-exposed mouse died.

Described implementations help to reveal the essence of the invention and allow adapting the invention to different uses, modify it, without deviating from the invention, by an equivalent replacement tools and materials when performing the various functions described in the application.

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1. A device for inducing apoptosis of abnormal cells in vitro and in vivo, containing means for generating a constant magnetic fields that permeate the working environment, in which the effect on the pathological cells or organs containing abnormal cells, and means for generating in a production environment, low-frequency electromagnetic fields in addition to the constant magnetic field, characterized in that the device further comprises means for modulating the constant magnetic fields associated with the means for their generation, establishing magnetic flux of the permanent magnetic fields from 1 to 100 MT in accordance with a given function of time, and means for modulating low-frequency electromagnetic fields associated with the means for generating the sets of low-frequency electromagnetic fields in accordance with a given function of the amplitude of the magnetic induction from 1 to 100 MT and frequencies from 1 to 1000 Hz from time to time.

2. A device for inducing apoptosis of abnormal cells in vitro and in vivo, containing tools for gene the purpose of the constant magnetic field, chilly working environment, in which the effect on the pathological cells or tissue containing abnormal cells, characterized in that it also includes means for modulating the constant magnetic fields associated with the means of their generation, establishing magnetic flux of the permanent magnetic fields from 1 to 100 MT in accordance with a given function of time.

3. A device for inducing apoptosis of abnormal cells in vitro and in vivo, characterized in that it also includes means for generating a low-frequency electromagnetic fields in the working environment in which these impact on pathological cells or tissue containing abnormal cells, and means for modulating low-frequency electromagnetic fields associated with the means for generating the sets of low-frequency electromagnetic fields in accordance with a given function of the amplitude of the magnetic induction in the range 1 - 100 MT and a frequency in the range 1 - 100 Hz from time to time.

4. The device according to claim 1 or 2, characterized in that the means for modulating the constant magnetic fields include software that establishes a magnetic induction according to the set specified with step-by-step interval of values of IC1, Ic2,...ICnfor appropriate temporary intervalof 1T2,...Tn.

5. The device according to claim 1 or 3, characterized in that the means for modulating low-frequency electromagnetic fields include software that sets the amplitude of the magnetic induction according to the set specified with step-by-step interval of values of IEN, IEN,...Iencpfor the respective time intervals T1T2,...Tn.

6. The device according to claim 1 or 3, characterized in that the means for modulating low-frequency electromagnetic fields include software tools that set the frequency according to the set specified with step-by-step interval of values of f1f2,...fnfor the respective time intervals T1T2,...Tnand named the frequency range 10 - 100 Hz.

7. The device according to claim 1, characterized in that the means for modulating the constant magnetic fields and low-frequency electromagnetic fields include software that sets the value of the constant magnetic fields and low-frequency electromagnetic field according to the set specified with step-by-step interval of values of IC1/IEN,, IC2,/IEN,...ICn/Iencpfor the respective time intervals T1T2,...Tn.

8. The device according to claim 7, characterized t is m, what software install permanent magnetic fields and low-frequency electromagnetic field according to the total magnetic induction in the range of 1 to 30 MT and the value of the constant magnetic fields and low frequency electromagnetic fields of 0.1 - 10, respectively.

9. The device according to claim 7, characterized in that the software install permanent magnetic fields and low-frequency electromagnetic field according to the total magnetic induction in the range 1 - 10 MT and the value of the constant magnetic fields and low frequency electromagnetic fields of 0.5 - 5, respectively.

10. Device according to any one of claims 4 to 9, characterized in that the software set intervals, 1 - 40 minutes

11. Device according to any one of claims 1 to 10, characterized in that at least one part of the working environment is limited by the walls, permeable to named fields.

12. Device according to any one of claims 1 to 11, characterized in that the means for creating a constant magnetic fields and/or low-frequency electromagnetic fields include at least first and second inductors, respectively surrounding at least part of the working environment, and means for modulating supply coils, respectively, direct and alternating current.

13. Device according to any one of claims 1 to 11, characterized those who, means for generating a constant magnetic fields and/or low-frequency electromagnetic fields include at least first and second coils, located coaxially, the working environment is located between the first and second coil, and means for modulating supply to the said coils, respectively, direct and alternating currents.

14. Device according to any one of claims 1 to 13, characterized in that it includes means providing within the working environment the generation of a constant electric field or an alternating electric field of low frequency up to 1000 Hz with an intensity of up to 20 kV/m

15. The method of use of the constant magnetic field and a variable low-frequency nonthermal electromagnetic fields for the induction of apoptosis by modifying gene mutant p53, characterized in that the permanent magnetic field and low-frequency electromagnetic fields have a magnetic induction of 1 - 100 MT and represent different sequences of constant magnetic fields and/or low-frequency electromagnetic fields, i.e. constant magnetic field, followed by low-frequency electromagnetic fields, low frequency electromagnetic fields, followed by a permanent magnetic field, a permanent magnetic field and low-frequency electromagnetic field, the action is either jointly, as well as permanent magnetic fields and low-frequency electromagnetic field, acting separately, with the low-frequency electromagnetic fields have a frequency of 1 to 1000 Hz.

16. The method according to item 15, characterized in that, along with the permanent magnetic fields and low-frequency electromagnetic fields used chemicals.

17. The method according to item 15, wherein the sequence of the constant magnetic fields and/or low-frequency electromagnetic fields are established at intervals of time T1T2,...Tnmoreover , the magnetic induction of the permanent magnetic fields and/or low-frequency electromagnetic fields in each time interval is set at a constant level values of IC1, IC2,...ICn; IEN, IEN,...Iencpand IC1/IEN, IC1/IEN,...ICn/Iencprespectively.

18. The method according to item 15, wherein the constant magnetic field and low-frequency electromagnetic fields are set with a total magnetic induction in the range of 1 to 30 MT, respectively, the ratio of the constant magnetic fields and low frequency electromagnetic fields of 0.1 - 10.

19. The method according to item 15, wherein the constant magnetic field and low-frequency electromagnetic fields are set with a total magnetic, indukti is in the range 1 - 10 MT, respectively, the ratio of the constant magnetic fields and low frequency electromagnetic fields of 0.5 to 2.5.



 

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FIELD: medicine; medical engineering.

SUBSTANCE: method involves acting upon pathological cells and tissues with constant and/or low frequency alternating electromagnetic field of 1 to 100 mTesla units large intensity and frequency of 1 to 1000 Hz. The device has means for producing constant and/or low frequency alternating electromagnetic fields permeating working space.

EFFECT: enhanced effectiveness in treating pathological tissues without harmfully influencing healthy cells.

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FIELD: medical engineering.

SUBSTANCE: device has three concentric inductance coils arranged in mutually perpendicular relation to each other, control unit producing signals for controlling feeding voltage of winding of each inductance coil. The control unit has PC with software installed and interface unit and three-channeled power amplifier. The PC is connected to the power amplifier for controlling voltage on each coil. The power amplifier is used for supplying voltage to coil windings. Coil windings are arranged turn-by-turn in a way that turns of the same number intersect.

EFFECT: enhanced effectiveness of treatment.

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FIELD: medicine.

SUBSTANCE: method involves treating organism with rotating magnetic field. Action mode I has magnetic field intensity of 1.7-3.4 mTesla units giving comfortable organism state. Mode II has magnetic field intensity of 2.9-5.8 mTesla units. Mode III has magnetic field intensity of 0.6-2.3 mTesla units giving discomfort organism state. The treatment is spread over the whole organism by applying modes I-III in any succession order with mode I selected at the beginning and end. Exposure time is equal to 5-10 min in each mode depending on initial organism state. The number of sessions is selected from 3 to 20 in manner individual for each organism.

EFFECT: enhanced effectiveness of treatment.

3 dwg, 1 tbl

FIELD: medicine; medical engineering.

SUBSTANCE: method involves applying variable electromagnetic field excited by amplitude-modulated signal having carrier frequency in 100-1000 kHz bandwidth and modulating frequency in 10-8000 kHz bandwidth, and irradiation with low intensity quasi-monochromatic radiation of visible and proximal infrared wavelength bandwidth with small spectrum width. Two action elements are used when performing the treatment. The first one is vaginal action element having inductive transmitter of resonant type emitting electromagnetic oscillations with carrier frequency belonging to 100-1000 kHz bandwidth amplitude-modulated in 10-8000 kHz bandwidth and low intensity quasi-monochromatic radiation of visible and proximal infrared wavelength bandwidth with small spectrum width. The treatment is carried out concurrently and in continuous mode. The device has power supply source, sinusoid signal oscillator and vaginal action element having dielectric casing. The casing encloses inductive transmitter winding. The sinusoid signal oscillator has active oscillator with inductive transmitter winding being concurrently used as inductive element of oscillatory circuit of the active oscillator. Low frequency oscillator is additionally available. Its output is connected to active oscillator circuit via modulator. Control tract has in series connected amplitude detector, DC amplifier and indicating unit. An additional supracutaneous treatment element has active oscillator, inductive transmitter winding concurrently serving as active oscillator inductance element, modulator, which input is connected to low frequency oscillator output and its output is connected to active oscillator circuit. The control tract has in series connected amplitude detector, DC amplifier and indicating unit. M photodiodes are connected in series via timer and acoustic indication element.

EFFECT: enhanced effectiveness of treatment.

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FIELD: medicine; medical engineering.

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EFFECT: enhanced effectiveness of treatment; accelerated healing process.

4 cl, 3 dwg

FIELD: medical engineering.

SUBSTANCE: device has cylinder cut along a plane passing through its axis and permanent electromagnet. The electromagnet is arranged in a way that one of its poles is in front of the first half of the cylinder and the other one is in front of the second half. Protrusions are available on the internal side of one of cylinder halves and openings are available on the other one. The openings and protrusions match each other in shape.

EFFECT: enhanced effectiveness in treating strictures with gradually increasing pressure without changing instrument; excluded traumatic urethral mucous membrane injuries.

1 dwg

FIELD: medical engineering.

SUBSTANCE: device has power supply source, M-photodiodes, inductive radiator mounted on flat dielectric carcass, active oscillator, modulator and low frequency oscillator circuits. The radiator is mounted in dielectric casing having two parts. Low frequency oscillator has oscillator of continuously variable frequency in frequency bandwidth of 10-8000 Hz. The device has additional control tract. The tract has in series connected amplitude detector, Dc amplifier and indicator light-emitting diode, timer and acoustic indication unit.

EFFECT: wide range of functional applications.

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FIELD: medicine.

SUBSTANCE: method involves determining availability of acute or chronic inflammatory process, inflammation form and stage. Acute inflammatory process being detected in paranasal sinuses of exudative form in catarrhal stage, treatment with extraweak electromagnetic field is administered at the following frequencies: 1.5; 2.5; 2.9; 53.5; 57.0; 75.5; 910.0 Hz in combination with quantum therapy in red light matching the electromagnetic field and directed to nose area in frequency, daily, 4-5 sessions per course. Acute inflammatory process being detected in paranasal sinuses of exudative form in serous stage, treatment with extraweak electromagnetic field is administered at the following frequencies: 2.5; 2.9; 1.7; 1.75; 9.4; 57.0; 58.0; 73.5; 522.0; 4900.0 Hz in combination with quantum therapy in blue and green light matching the electromagnetic field and directed to nose area in frequency, daily, 5-6 sessions per course. Acute or chronic inflammatory process being detected in paranasal sinuses of exudative form in purulent stage, treatment with extraweak electromagnetic field is administered at the following frequencies: 2.5; 2.9; 57.0; 58.0; 53.5; 727.0; 787.0; 802.0; 880.0; 11000.0 Hz in combination with quantum therapy in blue, green and red light matching the electromagnetic field and directed to nose area in frequency, daily, 6-7 sessions per course.

EFFECT: enhanced effectiveness of treatment.

FIELD: medical engineering.

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2 cl, 1 dwg

FIELD: medicine.

SUBSTANCE: physiotherapeutic apparatus has power supply, case with cap inside which M light-guides of one type and P light-guides of the other type are disposed. Device also inductive radiator with winding disposed onto flat dielectric frame, auto-generator circuit, modulator, low frequency signal generator and control circuit. Base with light-guides is installed in center of frame of inductive radiator. M light-guides are connected in series with power supply by means of first two-terminal two-positioned switch. P light-guides are connected in series with power supply by means of second two-terminal two-position switch. Control circuit has amplitude oscillator connected in series with dc amplifier and light-guide/indicator. Input of control circuit is connected with active oscillator circuit. Power circuits of active oscillator circuits, modulator, low frequency signal oscillator and control circuit are connected with power supply by means of third two-terminal two-position switch.

EFFECT: widened operational capabilities.

3 dwg

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