CN106621050A - Power frequency electromagnetic field generating device having tumor inhibiting effect and loading method - Google Patents
Power frequency electromagnetic field generating device having tumor inhibiting effect and loading method Download PDFInfo
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Abstract
The invention provides a power frequency electromagnetic field generating device having a tumor inhibiting effect. The power frequency electromagnetic field generating device is composed of a power frequency power supply, two auto-transformers, two diode bridges, two groups of coils, a radiation part, a voltage detection part, a magnetic field detection part, a temperature measuring part and a terminal computer. According to the device, on the basis of the power frequency 50/60Hz power supply, voltage is continuously changed by virtue of the DC and AC auto-transformers, so that superposed static and alternating electromagnetic fields are generated in the radiation part. A loading method of the electromagnetic fields having the tumor inhibiting effect has a certain extensibility, the frequency is within a range of 30-300Hz, the intensity ratio of the static electromagnetic field to the alternating electromagnetic field is at 0.5-2.5 and total intensity is 1-10mT. The invention provides the special power frequency electromagnetic field generating device and the loading method; the generating device is reasonable in design, capable of acting on organisms, noninvasive and low in toxic and side effects, and the generating device has the obvious inhibiting effect on a plurality of tumors; and due to the action of the magnetic fields, the content of superoxide radicals in cells can be increased, and autophagy is promoted.
Description
Technical field
The invention belongs to physics, biology and medical science crossing domain, are related to medicine equipment, especially tumor therapeutic is electric
Magnetic field.
Background technology
Low frequency electromagnetic field frequencies range within 300 kilo hertzs (kHz), not with fuel factor but with biological effect, its mechanism
Still fail to understand, may with act on charged macromolecular and change its functional status it is relevant.Magnetic field to the inhibitory action of tumour
There is wide coverage (referring to Artacho-Cord ó n Francisco etc., 2013, Int J Mol Sci；Tofani S, 2015,
Curr Topics in Med Chem).European Union and U.S. FDA approved tumor therapeutic magnetic field generating device(with
Se Lie Novocure companies) for the clinical treatment of polymorphy gliablastoma, the technical characteristic is to adopt 100-
The magnetic field (referring to United States Patent (USP) US8019414, US8019414, US8406870) of 300kHz.It is also proposed in Chinese patent many
The magnetic field loading method with tumor inhibitory effect is planted, including the high-intensity magnetic field more than 0.4 tesla (T) (referring to Chinese special
Sharp 90100152.X) and pulsed magnetic field (referring to Chinese patent 94111668.9).In the research and application in the field, electromagnetic field
Self attributes are different from effective object, mode, including the parameter such as time, intensity, pulse frequency, carrier frequency, interval time is not
Together, different biological effects can be produced.And the biophysical effect mechanism in magnetic field is complicated, not single molecular pathway is acted as
With, therefore to the biological effect in magnetic field, including the effect of suppression tumour cell, ununified and complete understanding.Specifically
Effect need to carry out examination checking for certain specific magnetic field load mode and specific effective object.
There is a kind of special load mode, the alternation magnetic that the static magnetic field that direct current is produced is produced with ultralow frequency alternating current
Field superposition.In theory, impact of this superposition magnetic field to free radical lone pair electrons singlet-triplet state transition is maximum, may
Increase energy level transition probability, extend the half-life of free radical, so as to expand its biological effect (referring to Tofani S, 1999,
Physica Medica；Barnes FS etc., 2015, Bioelectromagnetics).
Oncotherapy at present is generally using the operative treatment and noninvasive chemicotherapy of invasive.Radiotherapy can be general or office
Portion's property.Chemotherapy be divided into targeting with it is non-targeted.For chemicotherapy opposing and currently without the case of efficient targeting medicine, clinically
Urgently find new treatment method, especially noninvasive, low toxicity side effect method.Animals and humans are acted in low frequency electromagnetic field
During, not yet find obvious general toxic and side effect (referring to Tofani S etc., 2002, IEEE Transactions
on Plasma Science；Ronchetto F etc., 2004, Bioelectromagnetics).
Cell autophagy has an important physiological pathology function, and the good allusion quotation in big corner is obtained 2016 because of the pioneering contribution in this field
Nobel's medical science is encouraged with physiology.Autophagy determines with double-edged sword effect to cell fate, can help cell survival or induce thin
Born of the same parents' apoptosis is the important biomolecule feature and therapy target of malignant tumour (referring to White E etc., 2009, Clin Caner
Res；Hanahan D etc., 2011, Cell；Booth LA etc., 2014, Cell Signal).Low frequency magnetic field has inducing cell certainly
The effect (referring to Marchesi N etc., 2014, Cellular Physiology) bitten, but how to affect in tumour cell then
Survival and death, and concrete molecular mechanism, are not yet expressly understood that.
The content of the invention
It is an object of the invention to provide a kind of power frequency electromagnetic field generating means with tumor inhibition effect, is a kind of ultralow
Frequently, low-intensity, may act on organism and produces the field generator for magnetic and load mode of particular biological effect, the device and
Loading method has in vivo and in vitro tumor inhibition effect.
The present invention provides a kind of power frequency electromagnetic field generating means, by power frequency supply, the first auto-transformer (exchange AC), the
Two auto-transformers (direct current DC), the first diode bridge, the second diode bridge, first group of coil, second group of coil, irradiation
Part, voltage check part, magnetic field detection part, temperature-measuring part and terminal computer composition, power frequency supply respectively with the first self coupling
One end connection of transformer and the second auto-transformer, the other end of the first auto-transformer and the second auto-transformer respectively with
First diode bridge and the second diode bridge connect, and the other end of the first diode bridge is connected with first group of coil, the
The other end of two diode bridges is connected with second group of coil, and irradiation part is placed between first group of coil and second group of coil,
Voltage check part one end is connected respectively with first group of coil and second group of coil, and the other end is connected with terminal computer, magnetic field inspection
Each one end is connected respectively with irradiation part for survey part and temperature-measuring part, and the respective other end is connected respectively with terminal computer.
Irradiation part has two kinds, and the first is cell irradiation platform, for Tissue Culture Plate and culture dish design, by dividing plate
First, dividing plate two and the objective table between two dividing plates are constituted, and Tissue Culture Plate is positioned on objective table, dividing plate one
It is 12.9 centimetres with the spacing of dividing plate two, objective table height adjustable, it is ensured that Tissue Culture Plate bottom is located at irradiation zone center position.
Second irradiation part is animal irradiation platform, mainly for mouse design, by dividing plate one, dividing plate two and positioned at two every
Between plate 6 animal irradiation cells composition, constitute animal irradiation cell part include front apron, backboard, central dividing plate,
And cell dividing plate one, cell dividing plate two, cell dividing plate three, cell dividing plate four, wherein front apron and backboard bore multiple apertures
Used as air-vent, in order to avoid animal occurs anoxic, dividing plate one and the spacing of dividing plate two are 12.9 centimetres, and 6 irradiation cells are located at irradiation
Region center position, often taking turns irradiation can process 6 mouse.The material selection transparent resin plate of irradiation part, can clean and purple
Outer sterilization.
It is a further object to provide the method for carrying out magnetic field loading using described device, by following steps reality
It is existing：
Using power frequency 50/60Hz power supply, voltage is constantly changed with auto-transformer is exchanged by direct current, so as in irradiation portion
Part inside produces the static state (from direct current) and alternation (from exchanging) electromagnetic field of superposition；The concrete voltage schemes that change are with loading
Magnetic field intensity is as follows：It is divided into T1-T8 totally 8 time periods, T1 time periods DC voltage is 3 millivolts (mV), produces static electromagnetic field
Intensity is 2.97 milli teslas (mT), and alternating voltage mean value is 1.5mV, and it is 1.48mT to produce alternating electromagnetic field mean intensity；
T2 time periods DC voltage is 4mV, and it is 3.95mT to produce static electromagnetic field intensity, and alternating voltage mean value is 2.5mV, produces friendship
Electromagnetic Field mean intensity is 2.47mT；T3 time periods DC voltage is 3mV, and it is 2.97mT to produce static electromagnetic field intensity, is handed over
Stream average voltage is 1.5mV, and it is 1.48mT to produce alternating electromagnetic field mean intensity；T4 time periods DC voltage is 4mV, is produced
Static electromagnetic field intensity is 3.95mT, and alternating voltage mean value is 2.5mV, and it is 2.47mT to produce alternating electromagnetic field mean intensity；
T5 time periods DC voltage is 3mV, and it is 2.97mT to produce static electromagnetic field intensity, and alternating voltage mean value is 1mV, produces alternation
Electromagnetic field mean intensity is 1.08mT；T6 time periods DC voltage is 4mV, and it is 3.95mT to produce static electromagnetic field intensity, is exchanged
Average voltage is 1.5mV, and it is 1.48mT to produce alternating electromagnetic field mean intensity；T7 time periods DC voltage is 3mV, is produced quiet
State electromagnetic field intensity is 2.97mT, and alternating voltage mean value is 1mV, and it is 1.08mT to produce alternating electromagnetic field mean intensity；During T8
Between section DC voltage be 4mV, it is 3.95mT to produce static electromagnetic field intensity, and alternating voltage mean value is 1.5mV, produces alternation electricity
Magnetic field mean intensity is 1.48mT；Irradiation zone inside diverse location total magnetic intensity is identical, T1 to T8 time period average magnetic fields
Intensity is 5.5mT；The T1-T8 time periods change every time voltage chronic exposure 3.5-10 minutes, often take turns irradiation time cumulation 30-90 point
Clock, daily irradiation 2-4 wheels, for cell and animal, daily total exposure time is 60-120 minutes.
The electromagnetic field with tumor inhibitory effect of present invention description, its frequency is static (straight in the range of 30-300Hz
Stream) with alternation (exchange) electromagnetic field overall strength in the range of 1-10mT, static state (direct current) and alternation (exchange) electromagnetic field intensity ratio
Value is in the range of 0.5-2.5.
The magnetic fields time is that each time period changes every time voltage chronic exposure 3.5-10 minutes, often takes turns exposure time
30-90 minutes, for cell and animal, daily total exposure time is 60-120 minutes, and this device and loading method are to kinds of tumors
Significant inside and outside inhibitory action is produced, magnetic fields can increase intracellular superoxide radical content and promote cell autophagy.
By electromagnetic field generator and load mode described in the present invention, significant inside and outside is produced to kinds of tumors
Inhibitory action, magnetic fields can increase intracellular superoxide radical content and promote cell autophagy.In cell injuring model and
In cell viability experiment, show that produced magnetic field can suppress kinds of tumor cells to grow, including the nephroblastoma, nerve are female
The cells such as cytoma, breast cancer, colorectal cancer；In testing in vitro, can effectively suppress the growth of colorectal cancer and breast cancer
And transfer.
The present invention has the advantages that：(1) device is reasonable in design, easy to implement using unique power frequency；(2) will
The alternating magnetic field that the static magnetic field that direct current is produced is produced with alternating current is superimposed, and can affect the free radical in magnetic field, expands its life
Thing effect；(3) magnetic field intensity is uniform in irradiation part；(4) it is easy to operate, it is capable of achieving loading scheme by changing voltage；(5)
Associated voltage, magnetic field intensity, temperature that irradiation components interior produces magnetic field are continued to monitor, it is highly controllable；(6) it is placed in irradiation part
Interior organism is disturbed little, hurtless measure；(7) there is significant inhibitory action to kinds of tumors, and toxic and side effect is low；(8)
There is facilitation to cell autophagy.
Description of the drawings
Fig. 1 is electromagnetic field generator structural representation of the present invention.
Fig. 2 is one of apparatus of the present invention irradiation part, the cell irradiation platform schematic diagram being placed in magnetic field.
Fig. 3 is the two of apparatus of the present invention irradiation part, the animal irradiation platform schematic diagram being placed in magnetic field.
Fig. 4 is the major parameter of electromagnetic field load mode of the present invention, i.e., direct current (DC) with exchange (AC) voltage change scheme
With the corresponding electromagnetic field intensity for producing, alternating voltage is represented with mean value in figure.
Fig. 5 is inhibition figure of the present invention for nephroblastoma cell G401.
Fig. 6 is inhibition figure of the present invention for neuroblastoma cell CHLA255.
Fig. 7 is inhibition figure of the present invention for tumor growth Colon and rectum cell WiDr.
Fig. 8 is inhibition figure of the present invention for breast cancer cell MBA-MD-435 Lung metastases.
Fig. 9 is the design sketch that electromagnetic field changes superoxide radical content in nephroblastoma cell G401 in the present invention.
Figure 10 is the schematic diagram that electromagnetic field suppresses tumour cell G401 and CHLA255 in the present invention, including HRP chemiluminescences
Faxian shadow autophagy GAP-associated protein GAP band.
Specific embodiment
With reference to the accompanying drawings and examples the present invention will be further described.
Embodiment 1, a kind of power frequency electromagnetic field generating means with tumor inhibition effect that the present invention is provided, by a work
Frequency power 1, the first auto-transformer 2 (exchange AC), the second auto-transformer 3 (direct current DC), the first diode bridge 4, second
Diode bridge 5, first group of coil, 6, second group of coil 7, irradiation part 8, voltage check part 9, magnetic field detection part 10, survey
Warm part 11 and terminal computer 12 are constituted, and power frequency supply 1 is respectively with the one of the first auto-transformer 2 and the second auto-transformer 3
End connection, the other end of the first auto-transformer 2 and the second auto-transformer 3 respectively with the first diode bridge 4 and the 2nd 2
Pole pipe electric bridge 5 connects, and the other end of the first diode bridge 4 is connected with first group of coil 6, the second diode bridge 5 it is another
End is connected with second group of coil 7, and irradiation part 8 is placed between first group of coil 6 and second group of coil 7, voltage check part 9 one
End is connected respectively with first group of coil 6 and second group of coil 7, and the other end is connected with terminal computer 12, the He of magnetic field detection part 10
The respective one end of temperature-measuring part 11 is connected respectively with irradiation part 8, and the respective other end is connected respectively with terminal computer 12.
Irradiation part 8 has two kinds, the first be cell irradiation platform, for Tissue Culture Plate and culture dish design, by every
Plate 1, dividing plate 2 14 and the objective table between two dividing plates 15 are constituted, Tissue Culture Plate be positioned over objective table it
On, dividing plate 1 and the spacing of dividing plate 2 14 are 12.9 centimetres, objective table height adjustable, it is ensured that Tissue Culture Plate bottom is located at irradiation
Region center position.
Second irradiation part 8 is animal irradiation platform, mainly for mouse design, by dividing plate 1, dividing plate 2 14, with
And 6 animal irradiation cells 16 between two dividing plates are constituted, constituting the part of animal irradiation cell 16 includes front apron
18th, backboard 19, central dividing plate 20 and cell dividing plate 1, cell dividing plate 2 22, cell dividing plate 3 23, cell dividing plate four
24, wherein front apron 18 and backboard 19 bores multiple apertures 17 as air-vent, so as not to animal occur anoxic, dividing plate 1 and every
The spacing of plate 2 14 is 12.9 centimetres, and 6 irradiation cells 16 are located at irradiation zone center position, and often taking turns irradiation can process 6 mouse.
The material selection transparent resin plate of irradiation part 8, can clean and ultraviolet disinfection.
Concrete magnetic field load mode takes scheme as shown in Figure 4, using power frequency 50/60Hz power supply, by direct current with exchange
Auto-transformer constantly changes voltage, so as to produce the electromagnetic field that static state is superimposed with alternating field in irradiation components interior；Specifically change
Time variant voltage scheme is as follows with the magnetic field intensity for producing：It is divided into T1-T8 totally 8 time periods, T1 time periods DC voltage is 3mV, is produced
Raw static electromagnetic field intensity is 2.97mT, and alternating voltage mean value is 1.5mV, produces alternating electromagnetic field mean intensity and is
1.48mT；T2 time periods DC voltage is 4mV, and it is 3.95mT to produce static electromagnetic field intensity, and alternating voltage mean value is
2.5mV, it is 2.47mT to produce alternating electromagnetic field mean intensity；T3 time periods DC voltage is 3mV, produces static electromagnetic field intensity
For 2.97mT, alternating voltage mean value is 1.5mV, and it is 1.48mT to produce alternating electromagnetic field mean intensity；T4 time period direct currents
Press as 4mV, it is 3.95mT to produce static electromagnetic field intensity, and alternating voltage mean value is 2.5mV, produces alternating electromagnetic field averagely strong
Spend for 2.47mT；T5 time periods DC voltage is 3mV, and it is 2.97mT to produce static electromagnetic field intensity, and alternating voltage mean value is
1mV, it is 1.08mT to produce alternating electromagnetic field mean intensity；T6 time periods DC voltage is 4mV, produces static electromagnetic field intensity and is
3.95mT, alternating voltage mean value is 1.5mV, and it is 1.48mT to produce alternating electromagnetic field mean intensity；T7 time period DC voltages
For 3mV, it is 2.97mT to produce static electromagnetic field intensity, and alternating voltage mean value is 1mV, produces alternating electromagnetic field mean intensity and is
1.08mT；T8 time periods DC voltage is 4mV, and it is 3.95mT to produce static electromagnetic field intensity, and alternating voltage mean value is
1.5mV, it is 1.48mT to produce alternating electromagnetic field mean intensity；Irradiation zone inside diverse location total magnetic intensity is identical, and T1 is extremely
T8 time periods average field strength is 5.5mT；The T1-T8 time periods change every time voltage chronic exposure 3.5-10 minutes, per spoke
According to time cumulation 30-90 minutes, daily irradiation 2-4 wheels, for cell and animal, daily total exposure time is 60-120 minutes.
Embodiment 2, the present invention acts on the nephroblastoma cell G401 of culture, using Fig. 1 and Fig. 2 shown devices, presses
Cell is processed according to scheme described in Fig. 4, voltage is changed every time and is acted on 3.5 minutes, often taken turns about 30 minutes altogether action time, processed daily
4 take turns, totally 2 hours Electromagnetic Field Irradiation time；The articles holding table that negative control group is placed in cell between the coil for not loading electromagnetic field,
Time is identical；Positive control is cis-platinum (Jiangsu Haosen Pharmaceutical Co., Ltd), and administration concentration is 500nM, and 1 is administered daily
It is secondary.
Specific implementation process is：
Clone and cultural method：Nephroblastoma cell G401, this cell is attached cell, with containing 10% hyclone
McCoy medium cultures in 37 DEG C of incubators, containing 5% carbon dioxide and saturated steam；
Capital equipment：Power frequency electromagnetic field generating means, cell culture incubator, viable count instrument, multi-function microplate reader；
Main agents：CCK8 kits, fluorescence probe DCFH-DA, BCA kits, 40% acrylamide gel storage liquid,
Tris alkali, TEMED, SDS, protease inhibitors, protein standard substance, HRP chemistry is given out light detection kit；
Antibody：One it is anti-include LC3, GAPDH, p62, it is anti-that anti-mouse is coupled two with anti-rabbit HRP；
Cell viability is tested：Nephroblastoma cell G401 is inoculated in into 6 orifice plates with the density in 1 × 10 5/holes of ∧, often
3 multiple holes of individual treatment conditions, control group, irradiation group and cis-platinum group are processed respectively according to such scheme, are collected daily at the previous day
The cell of reason, with viable count instrument meter number, according to reading cell number is calculated, and makes cell growth curve and electromagnetic field to thin
The suppression curve of born of the same parents；
Statistical analysis：Every kind of radiation parameter is repeated 3 times, every time 3 multiple holes, is as a result represented with mean value ± standard error, knot
Fruit is cooked T inspections, with P<0.05 used as with significant difference index；
As shown in figure 5, left hand view is cell growth curve, wherein X-axis is magnetic field radiation number of days, and Y-axis is cell number, curve
One is cellular control unit growth curve, and curve two is irradiation group cell growth curve, and curve three is positive control cis-platinum group cell
Growth curve；Right part of flg is Carbazole alkaloid curve, and wherein X-axis is magnetic field radiation number of days, and Y-axis is tumour inhibiting rate (%), and curve one is
Irradiation group cell inhibitory rate curve, curve two is cis-platinum group cell inhibitory rate curve；Result shows irradiation group cell propagation in figure
Slow down, inhibiting rate increases；
Superoxide radical assay：Nephroblastoma cell G401 is inoculated in into 96 with the density in 1 × 10 4/holes of ∧
Orifice plate, 3 multiple holes of each treatment conditions, irradiation group is processed according to scheme described in Fig. 4, is often taken turns 30 minutes time, and control group is placed on
The articles holding table between the coil of electromagnetic field is not loaded, a coprocessing 4 is taken turns, and exposure time amounts to 120 minutes, after often wheel process terminates
It is incubated 20 minutes in incubator with fluorescence probe DCFH-DA (10 μM), the probe for being introduced into cell is washed away with PBS after incubation,
Detected with multi-function microplate reader, excitation wavelength 488nm, absorbing wavelength 525nm, each group fluorescence intensity level is read respectively；Such as Fig. 9 institutes
Show, wherein X-axis be the magnetic field radiation time, Y-axis for intracellular probes fluorescence intensity, i.e. superoxide radical relative amount, curve
One is the relative amount of superoxide radical in cellular control unit, and curve two contains for the relative of the intracellular superoxide radical of irradiation group
Amount, result shows that the intracellular superoxide radical of irradiation group has increased after the wheel irradiation of 30 minutes is received than control group in figure
Height, and be continued for, the level being close to control group was down to when 120 minutes, this result illustrates this magnetic field load mode
Really intracellular free radicals content can be improved, and it is effective rapid, and the time is within an hour.
Immuning hybridization is tested：After cell process after centrifugation, PBS washings, the RIPA buffer solutions for being suspended in precooling (add albumen
Enzyme and phosphorglase inhibitor), kit BCA methods are carried out after protein quantification, and albumen is separated by electrophoresis with PAGE gel, per hole
Applied sample amount is 20 μ g, is carried out with appropriate antibody after immuning hybridization, HRP chemoluminescence methods development protein band, as shown in Figure 10,
P62 is reduced after irradiation, and LC3-II increases, and illustrates the inducible G401 cell autophagies of irradiation.
Embodiment 3, the present invention acts on the neuroblastoma cell CHLA255 of culture, fills using shown in Fig. 1 and Fig. 2
Put, according to scheme described in Fig. 4 cell is processed, voltage is changed every time and is acted on 3.5 minutes, often take turns about 30 minutes action time, daily
Process 4 to take turns, totally 2 hours Electromagnetic Field Irradiation time；Negative control group is placed in cell between the coil for not loading electromagnetic field
Articles holding table same time；Positive control is cis-platinum (Jiangsu Haosen Pharmaceutical Co., Ltd), and administration concentration is 500nM, daily
It is administered once.
Specific implementation process is：
Clone and cultural method：Neuroblastoma cell CHLA255, this cell is attached cell, with containing 10% tire
The IMDM medium cultures of cow's serum in 37 DEG C of incubators, containing 5% carbon dioxide and saturated steam；
Capital equipment：Power frequency electromagnetic field generating means, cell culture incubator, viable count instrument；
Main agents：CCK8 kits, BCA kits, 40% acrylamide gel storage liquid, Tris alkali, TEMED,
SDS, protease inhibitors, albumen loading standard product, HRP chemistry is given out light detection kit；
Antibody：One it is anti-include LC3, GAPDH, p62, it is anti-that anti-mouse is coupled two with anti-rabbit HRP；
Cell viability is tested：Neuroblastoma cell CHLA255 is inoculated in into 6 holes with the density in 1 × 10 5/holes of ∧
Plate, 3 multiple holes of each treatment conditions, control group, irradiation group and cis-platinum group are processed respectively according to such scheme, collect previous daily
Its cell for processing, with viable count instrument meter number, according to reading cell number is calculated, and makes cell growth curve and electromagnetic field
Suppression curve to cell；
Statistical analysis：Every kind of radiation parameter is repeated 3 times, every time 3 multiple holes, is as a result represented with mean value ± standard error, knot
Fruit is cooked T inspections, with P<0.05 used as with significant difference index；
As shown in fig. 6, left hand view is cell growth curve, wherein X-axis is magnetic field radiation number of days, and Y-axis is cell number, curve
One is cellular control unit growth curve, and curve two is irradiation group cell growth curve, and curve three is cis-platinum group cell growth curve；
Right part of flg is Carbazole alkaloid curve, and wherein X-axis is magnetic field radiation number of days, and Y-axis is tumour inhibiting rate (%), and curve one is irradiation group cell
Inhibiting rate curve, curve two is cis-platinum group cell inhibitory rate curve, and result shows that irradiation group cell propagation slows down in figure, inhibiting rate
Increase；
Immuning hybridization is tested：After cell process after centrifugation, PBS washings, the RIPA buffer solutions for being suspended in precooling (add albumen
Enzyme and phosphorglase inhibitor), BCA methods are carried out after protein quantification, and albumen is separated by electrophoresis with PAGE gel, per hole applied sample amount
For 20 μ g, carried out with appropriate antibody after immuning hybridization, HRP chemoluminescence methods development protein band, as shown in figure 9, after irradiation
P62 is reduced, and LC3-II increases, and illustrates the inducible CHLA255 cell autophagies of irradiation.
Embodiment 4, the present invention acts on the colorectal cancer cell WiDr of tumor growth,
Specific implementation process is：After nude mice by subcutaneous inoculation colon adenocarcinoma cell WiDr, irradiation group is used shown in Fig. 1 and Fig. 3
Device, according to scheme described in Fig. 4 animal is processed, and voltage is changed every time and is acted on 10 minutes, and it is 70 minutes often to take turns exposure time, daily
Irradiation 1 is taken turns.Control group is put into the animal irradiation platform without magnetic field simultaneously.Per day entry irradiation group with control group mice existence feelings
Condition.As shown in fig. 7, irradiation group survival time of mice significantly extends.
Embodiment 5, the present invention is directed to breast cancer cell MBA-MD-435,
Specific implementation process is：SCID mouse subcutaneous vaccination breast cancer cell MBA-MD-435, irradiation group uses Fig. 1 and Fig. 3
Shown device, according to scheme described in Fig. 4 animal is processed, and voltage is changed every time and is acted on 10 minutes, and it is 70 minutes often to take turns exposure time,
Process 1 daily to take turns, 5 days weekly, continuous 4 weeks.Negative control group is put into the irradiation platform for being not added with loaded magnetic field simultaneously.Positive control
Group gives endoxan, and dosage is 50mg/kg, and one time a day.Process in 4 weeks puts to death animal after terminating, and takes lungs and fixes, and pathology is cut
Piece, observes and counts Pulmonary metastasis focuses number and size, * * P<0.01,***P<0.001, as shown in figure 8, the transfer of irradiation group mouse
Stove number is substantially reduced, and effect is suitable with endoxan, while irradiation group MET is substantially reduced, the anti-transfer effect of irradiation is better than
Endoxan.
Claims (7)
1. a kind of power frequency electromagnetic field generating means with tumor inhibition effect, it is characterised in that by a power frequency supply (1),
First auto-transformer (2), the second auto-transformer (3), the first diode bridge (4), the second diode bridge (5), first
Group coil (6), second group of coil (7), irradiation part (8), voltage check part (9), magnetic field detection part (10), temperature-measuring part
(11) with terminal computer (12) constitute, power frequency supply (1) respectively with the first auto-transformer (2) and the second auto-transformer (3)
One end connection, the other end of the first auto-transformer (2) and the second auto-transformer (3) respectively with the first diode bridge
(4) connect with the second diode bridge (5), the other end of the first diode bridge (4) is connected with first group of coil (6), second
The other end of diode bridge (5) is connected with second group of coil (7), and irradiation part (8) is placed in first group of coil (6) and second group
Between coil (7), voltage check part (9) one end is connected respectively with first group of coil (6) and second group of coil (7), the other end
Be connected with terminal computer (12), magnetic field detection part (10) and temperature-measuring part (11) each one end respectively with irradiation part (8) phase
Even, the respective other end is connected respectively with terminal computer (12).
2. a kind of power frequency electromagnetic field generating means with tumor inhibition effect according to claim 1, it is characterised in that
Irradiation part (8) is divided into two kinds, and the first part (8) is cell irradiation platform, by dividing plate one (13), dividing plate two (14) and
Objective table (15) composition between two dividing plates, Tissue Culture Plate is positioned on objective table (15), dividing plate one (13) and every
The spacing of plate two (14) is 12.9 centimetres, objective table (15) height adjustable, it is ensured that Tissue Culture Plate bottom is being located at irradiation zone just
Middle position.
3. a kind of power frequency electromagnetic field generating means with tumor inhibition effect according to claim 1, it is characterised in that
Second irradiation part (8) is animal irradiation platform, by dividing plate one (13), dividing plate two (14) and between two dividing plates
6 animals irradiation cell (16) composition, constitute the part of animal irradiation cell 16 include front apron (18), backboard (19),
Central dividing plate (20) and cell dividing plate one (21), cell dividing plate two (22), cell dividing plate three (23), cell dividing plate four (24),
Wherein front apron (18) and backboard (19) bore multiple apertures (17) as air-vent, in case there is anoxic, dividing plate one in animal
(13), the spacing of dividing plate two (14) is 12.9 centimetres, and, positioned at irradiation zone center position, often taking turns irradiation can for 6 irradiation cells (16)
Process 6 mouse.
4. a kind of power frequency electromagnetic field generating means with tumor inhibition effect according to claim 1,2 or 3, its feature
It is, the material selection transparent resin plate of described irradiation part (8).
5. the method that according to claim 1 device carries out power frequency magnetic field loading, it is characterised in that by following steps reality
It is existing：
Using power frequency 50/60Hz power supply, voltage is constantly changed with auto-transformer is exchanged by direct current, so as in irradiation part
Portion produces the static and alternating electromagnetic field of superposition, and the concrete voltage schemes that change are as follows with the magnetic field intensity of loading：It is divided into T1-T8 to be total to
8 time periods, T1 time periods DC voltage is 3mV, and it is 2.97mT to produce static electromagnetic field intensity, and alternating voltage mean value is
1.5mV, it is 1.48mT to produce alternating electromagnetic field mean intensity；T2 time periods DC voltage is 4mV, produces static electromagnetic field intensity
For 3.95mT, alternating voltage mean value is 2.5mV, and it is 2.47mT to produce alternating electromagnetic field mean intensity；T3 time period direct currents
Press as 3mV, it is 2.97mT to produce static electromagnetic field intensity, and alternating voltage mean value is 1.5mV, produces alternating electromagnetic field averagely strong
Spend for 1.48mT；T4 time periods DC voltage is 4mV, and it is 3.95mT to produce static electromagnetic field intensity, and alternating voltage mean value is
2.5mV, it is 2.47mT to produce alternating electromagnetic field mean intensity；T5 time periods DC voltage is 3mV, produces static electromagnetic field intensity
For 2.97mT, alternating voltage mean value is 1mV, and it is 1.08mT to produce alternating electromagnetic field mean intensity；T6 time period DC voltages
For 4mV, it is 3.95mT to produce static electromagnetic field intensity, and alternating voltage mean value is 1.5mV, produces alternating electromagnetic field mean intensity
For 1.48mT；T7 time periods DC voltage is 3mV, and it is 2.97mT to produce static electromagnetic field intensity, and alternating voltage mean value is
1mV, it is 1.08mT to produce alternating electromagnetic field mean intensity；T8 time periods DC voltage is 4mV, produces static electromagnetic field intensity and is
3.95mT, alternating voltage mean value is 1.5mV, and it is 1.48mT to produce alternating electromagnetic field mean intensity.
6. method according to claim 5, it is characterised in that by auto-transformer, produce in irradiation components interior folded
Plus static and alternating electromagnetic field, irradiation zone inside diverse location total magnetic intensity is identical, in exposure time section plus loaded magnetic field
Excursion be 1.0-7.0mT, mean intensity is 5.5mT.
7. method according to claim 5, it is characterised in that the magnetic fields time is that each time period changes every time electricity
Pressure chronic exposure 3.5-10 minutes, often take turns exposure time 30-90 minute, for cell and animal, daily total exposure time is 60-
120 minutes.
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| CN201710066756.XA CN106621050A (en) | 2017-02-07 | 2017-02-07 | Power frequency electromagnetic field generating device having tumor inhibiting effect and loading method |
| PCT/IB2018/050765 WO2018172863A1 (en) | 2017-02-07 | 2018-02-07 | Apparatus for treating pathological cells |
| EP18711400.4A EP3579920A1 (en) | 2017-02-07 | 2018-02-07 | Apparatus for treating pathological cells |
| US16/483,862 US11364390B2 (en) | 2017-02-07 | 2018-02-07 | Apparatus for treating pathological cells |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
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| WO2018172863A1 (en) | 2017-02-07 | 2018-09-27 | Santi Tofani | Apparatus for treating pathological cells |
| CN108607162A (en) * | 2018-05-25 | 2018-10-02 | 浙江大学 | A kind of tumor suppression device of program control and use |
| CN113546321A (en) * | 2021-07-14 | 2021-10-26 | 上海理工大学 | Low-frequency magnetic field generating device for mechanical therapy and thermal therapy of tumors and using method thereof |
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| WO2018172863A1 (en) | 2017-02-07 | 2018-09-27 | Santi Tofani | Apparatus for treating pathological cells |
| US11364390B2 (en) | 2017-02-07 | 2022-06-21 | Santi Tofani | Apparatus for treating pathological cells |
| CN108607162A (en) * | 2018-05-25 | 2018-10-02 | 浙江大学 | A kind of tumor suppression device of program control and use |
| CN113546321A (en) * | 2021-07-14 | 2021-10-26 | 上海理工大学 | Low-frequency magnetic field generating device for mechanical therapy and thermal therapy of tumors and using method thereof |
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