PEMF Potentiates Doxorubicin-induced Late G2 Arrest in MDA-MB-231 Breast Cancer Cells.

BACKGROUND/AIM: Pulsed electromagnetic field (PEMF) stimulation enhances the efficacy of several anticancer drugs. Doxorubicin is an anticancer drug used to treat various types of cancer, including breast cancer. However, the effect of PEMF stimulation on the efficacy of doxorubicin and the underlying mechanisms remain unclear. Thus, this study aimed to investigate the effect of PEMF stimulation on the anticancer activity of doxorubicin in MDA-MB-231 human breast cancer cells. MATERIALS AND METHODS: MDA-MB-231 cells were seeded and allowed to incubate for 48 h. The cells were treated with doxorubicin, cisplatin, 5-fluorouracil, or paclitaxel for 48 h. Subsequently, the cells were stimulated with a 60-min PEMF session thrice a day (with an interval of 4 h between each session) for 24 or 48 h. Cell viability was assessed by trypan blue dye exclusion assay and cell-cycle analysis was analyzed by flow cytometry. Molecular mechanisms involved in late G2 arrest were confirmed by a western blot assay and confocal microscopy. RESULTS: MDA-MB-231 cells treated with a combination of doxorubicin and PEMF had remarkably lower viability than those treated with doxorubicin alone. PEMF stimulation increased doxorubicin-induced cell-cycle arrest in the late G2 phase by suppressing cyclin-dependent kinase 1 (CDK1) activity through the enhancement of myelin transcription factor 1 (MYT1) expression, cell division cycle 25C (CDC25C) phosphorylation, and stratifin (14-3-3σ) expression. PEMF also increased doxorubicin-induced DNA damage by inhibiting DNA topoisomerase II alpha (TOP2A). CONCLUSION: These findings support the use of PEMF stimulation as an adjuvant to strengthen the antiproliferative effect of doxorubicin on breast cancer cells.

Pulsed Electromagnetic Field Enhances Caffeic Acid Phenethyl Ester-induced Death of MCF-7 Breast Cancer Cells.

BACKGROUND/AIM: Caffeic acid phenethyl ester (CAPE) exerts anticancer effects against several cancer types, including breast cancer. Pulsed electromagnetic field (PEMF) improves the efficiency of some chemotherapeutic drugs. In this study, we examined the effects of PEMF stimulation on the anticancer activity of CAPE in MCF-7 breast cancer cells and the underlying signal transduction pathways. MATERIALS AND METHODS: MCF-7 cells were seeded and incubated for 24 h. Each of the drugs (5-fluorouracil, paclitaxel, gefitinib, or CAPE) was added to the cells on day 0. Then, cells were immediately stimulated with a 60-min PEMF session thrice a day (with 4-h interval between sessions) for 1-3 days. Cell death and viability were assessed by flow cytometry and trypan blue dye exclusion assay. Molecular mechanisms involved in cell death were confirmed by western blot assay. RESULTS: Compared with treatment with CAPE alone, co-treatment with CAPE and PEMF more strongly reduced the viability of MCF-7 cells, further increased the percentage of the sub-G1 population, poly (ADP-ribose) polymerase (PARP) cleavage, activation of apoptotic caspases, up-regulation of pro-apoptotic proteins, such as Fas cell surface death receptor (FAS) and BCL2 associated X, apoptosis regulator (BAX), and reduced the expression of anti-apoptotic proteins, such as BCL-2 apoptosis regulator (BCL-2), MCL-1 apoptosis regulator, BCL-2 family member (MCL-1), and survivin. PEMF stimulation also increased CAPE-induced phosphorylation of p53, and inhibition of p53 partially restored the PEMF-reduced viability of CAPE-treated MCF-7 cells. CONCLUSION: PEMF stimulation enhanced CAPE-induced cell death by activating p53, which regulates the expression of apoptosis-related molecules, subsequently activating the caspase-dependent apoptotic pathway in MCF-7 cells, suggesting that PEMF can be utilized as an adjuvant to enhance the effect of CAPE on breast cancer cells.

Evaluation of Atopic Dermatitis Improvement Caused by Low-Level, Low-Frequency Pulsed Electromagnetic Fields.

This study aimed to evaluate the effectiveness of using low-level, low-frequency pulsed electromagnetic field (LLLF_PEMF) stimulation to improve atopic dermatitis induced by 2,4-dinitrochlorobenzene (DNCB). Twenty 6-week-old hairless mice were randomly divided into Normal (n = 5), PEMF 15 Hz (n = 5), PEMF 75 Hz (n = 5), and Sham (n = 5) groups. Following the onset of atopic dermatitis symptoms, PEMF groups (15 and 75 Hz) were stimulated with LLLF_PEMF (15 mT) for 8 h per day for 1 week. Sensory evaluation analysis revealed a significant difference between the PEMF 15 Hz group and Sham group (P < 0.05), but these differences were not visually obvious. While both the PEMF and Sham groups had atopic dermatitis lesions, lesion size was significantly smaller in the two PEMF groups than in the Sham group (P < 0.001). Additionally, changes in epithelial thickness because of skin inflammation significantly decreased for both PEMF groups, compared with the Sham group (P < 0.001). In conclusion, these results suggest that PEMF stimulation in vivo triggers electro-chemical reactions that affect immune response. © 2022 Bioelectromagnetics Society.

Effect of pulsed electromagnetic fields stimulation on ischemic skin model.

This study aimed to evaluate the effect of Pulsed Electromagnetic Fields (PEMF) in improving blood flow reduction and tissue necrosis of ischemic animal induced by skin flap. In each experiment, twenty rats (280-320 g) were randomly divided into control group (n = 10) and PEMF (n = 10) group. All of the rats were performed skin flap in back. In the PEMF group, PEMF (1 Hz, 10 mT) was performed in each experiment. In Experiment-1 (n = 20), PEMF was performed for 90 minutes. In Experiment-2 (n = 20), additionally, a blocking film was inserted, and suture was performed to induce necrosis. PEMF was performed for 30 minutes each day for 7 days. As a result of Speckle-Flow Index (SFI) analysis, in the control group, blood flow continued to decrease immediately after the procedure. In the PEMF group, blood flow was remained constant after 30 minutes and increased after 60 minutes. The blood flow in a specific region substantially increased from the initial state. As a result of skin necrosis analysis, the progression rate in the PEMF group was slower than that of the control group. The rate of necrosis in the PEMF group decreased dramatically from the 6th day, and there was a statistically significant difference between the two groups at the 7th day (p < .05). In this study, it was confirmed that PEMF (1 Hz, 10 mT) has a blood flow improvement and skin tissue necrosis alleviation in the ischemic flap animal model.

Comparison of the Effects of Manual Acupuncture, Laser Acupuncture, and Electromagnetic Field Stimulation at Acupuncture Point BL15 on Heart Rate Variability.

The aim of this study was to compare the influences of manual acupuncture, laser acupuncture, and electromagnetic field stimulation on the autonomic nervous system. We monitored the heart rate variability before and after stimulation to check the influence on the autonomic nervous system. The heart rate variabilities at low frequency (LF; 0.04-0.15 Hz) and high frequency (HF; 0.15-0.4 Hz) were analyzed to acquire LF/HF ratio. Xinshu (BL15) was selected as the stimulation point. Methods included manual acupuncture with a 1-cm depth and laser acupuncture at a wavelength of 660 nm and output power of 50 mW. An electromagnetic field of 2 Hz and 460 gauss (46 mT) was chosen. The LF and the LF/HF ratio were found to be lower in the manual acupuncture and the electromagnetic field groups, but to be higher in the laser acupuncture group. The HF was found to be lower in the laser acupuncture group, but higher in the manual acupuncture and the electromagnetic field groups. In conclusion, we found that manual acupuncture and electromagnetic field stimulation at BL15 activated the parasympathetic nervous system, whereas laser acupuncture at BL15 activated the sympathetic nervous system.

Electromagnetic acupuncture to enhance the effects of manual acupuncture on recovery from muscle fatigue of the quadriceps.

The aim of this study was to investigate a new method of manual acupuncture that used a magnetic field to stimulate only one acupoint vertically. We developed an eight-channel electromagnetic acupuncture (EMA) system that uses a solenoid-type electrode to insert the manual acupuncture needle into a hole in an electrode. We used a manual acupuncture needle for magnetic induction in order to penetrate vertically and deeply into tissues. In order to confirm the usefulness of EMA, we investigated the effects of treatment on muscle fatigue after strenuous knee extension/flexion exercises that had been performed by three groups: the nonstimulation, the manual acupuncture, and the EMA groups. Electromyograms showed that the median frequency (MF) in the EMA group had rapidly recovered after 4 minutes (p = 0.608), but that the peak torque had not recovered to the normal state (p < 0.05). Thus, we confirmed that compared with manual acupuncture, EMA resulted in better recovery from muscle fatigue.

Numerical analysis of the change in skin color due to ecchymosis and petechiae generated by cupping: a pilot study.

Cupping is one of the various treatment methods used in traditional oriental medicine. Cupping is also used as a diagnostic method and it may cause skin hyperpigmentation. Quantitative measurements and analysis of changes in skin color due to cupping are critical. The purpose of this study is to suggest an optical technique to visualize and identify changes in skin color due to cupping. We suggest the following analysis methods: digital color spaces [red, green, and blue (RGB) and L∗a∗b], the Erythema Index (E.I.), and the Melanin Index (M.I.). For experiments, we selected and stimulated 10 acupoints at 80 kilopascals (kPa) per minute. The RGB and L∗a∗b color spaces were observed to be decreased (p < 0.05) after cupping. The E.I. and M.I. were observed to be increased significantly (p < 0.05) after cupping. To assess various changes in skin color, we observed the changes for 72 hours. We also obtained the color changes by using the recovery pattern during the recovery period (p < 0.01). We propose that this method can be useful for visual identification and as a way to improve the identification of skin color changes.

Use of pupil size to determine the effect of electromagnetic acupuncture on activation level of the autonomic nervous system.

Magnetic fields are widely considered as a method of treatment to increase the therapeutic effect when applied to acupoints. Hence, this study proposes a new method which creates significant stimulation of acupoints by using weak magnetic fields. We conducted this experiment in order to confirm the effect on the activation level of the autonomic nervous system by measuring pupil sizes in cases of stimulation by using manual acupuncture and electromagnetic acupuncture (EMA) at BL15. We selected 30 Hz of biphasic wave form with 570.1 Gauss. To confirm the biopotential by the magnetic flux density occurring in EMA that affected the activation of the autonomic nervous system, we observed the biopotential induced at the upper and the mid left and right trapezius. We observed a significant decrease in pupil size only in the EMA group (p < 0.05), thus confirming that EMA decreased the pupil size through activation of the parasympathetic nerve in the autonomic nervous system. Moreover, we confirmed that the amplitude of the biopotential which was caused by 570.1 Gauss was higher than ±20 µA. Thus, we can conclude that EMA treatment successfully activates the parasympathetic nerve in the autonomic nervous system by inducing a biotransformation by the induced biopotential.

Development and evaluation of a multi-frequency bioelectrical impedance analysis analyzer for estimating acupoint composition.

The purpose of this study was to suggest a new method of estimating acupoint compositions by using a multi-frequency bioelectrical impedance analysis (MF-BIA) method at 5 kHz, 50 kHz and 200 kHz within 2 cm of acupoints divided into local segments. To verify the system developed, we confirmed the stable occurrence of a constant current at every frequency, regardless of the impedance connected to the electrodes. Moreover, we found left and right distal bicep brachii aponeurosis to be identical by using ultrasound imaging, and we analyzed the repeatability of the findings by making 10 consecutive sets of measurements (p > 0.05). To evaluate the practical use of the acupoint composition, we used the MF-BIA analyzer to measure the left and right LU3, LU4, and LU9 at the lung meridian. We confirmed that the potentials generated were equal to the changes in the cell membrane function, which were caused by the applied frequency (p < 0.01). We also verified that the MF-BIA analyzer measurements corresponded to the acupoint components by comparing the left and right potentials generated (p > 0.05). Hence, we conclude that the MF-BIA analyzer can be used to estimate the acupoint composition based on the acupoint state.

Development and evaluation of a bio-ion measurement system on acupoints for meridian diagnosis.

The aim of this study was to develop a new method of measuring the meridian energy potential (MEP) at an acupoint with bioelectrical impedance in order to establish an objective meridian diagnosis. To confirm the reflection of the changes in body composition at acupoints, the changes in the MEP on the bladder meridian before, during and after cupping were observed. Additionally, to evaluate the usefulness of diagnosis, the MEP on the stomach meridian before and after a meal was measured. Significant increases were found at every acupoint during cupping and after eating a meal. It was confirmed that the MEP data reflected changes in body composition at every acupoint and that these changes were associated with functioning of the associated internal organs. In conclusion, we propose the usefulness of this method, which can indicate the state of acupoints.

Comparison of 2 methods of non-invasive treatment between transcutaneous electrical stimulation and pulsed electromagnetic field stimulation as replacement of invasive manual acupuncture.

The aim of this study was to find the non-invasive optimal alternative method for Manual Acupuncture. Existing researches had reported that Transcutaneous Electrical Acupoint Stimulation (TEAS) was an effective treatment method instead of manual acupuncture. In place of the TEAS, we suggested the Pulsed Electromagnetic Fields (PEMFs). Thus, we designed the PEMFs system which can stimulate only an acupoint. There have been no researches which reported therapeutic effect when stimulating at an identical acupoint by TEAS and PEMFs. Hence, this study investigated the therapeutic effect on the muscle fatigue after the strenuous knee extension/flexion exercise by two stimulations. We selected the stimulation method of both TEAS and PEMFs by using 2Hz biphasic rectangular wave pulse and pulse width 0.2ms. The magnetic flux was the 30.92mT (309.2gauss) at 2 Hz. The electromyogram (EMG) and the maximal voluntary contraction (MVC) at rectus femoris were measured. The Median Frequency (MF) at TEAS group was significantly effective at 6 minutes (p=0.499). The PEMFs group was recovered to the MF rapidly after 4 minutes (p=0.166). The results of the peak torque indicated that both non-stimulation group and TEAS group did not recover to the peak torque at pre-exercise during the recovery period (p<0.05). In contrast, the significant treatment effect of PEMFs group was found after 14 minutes (p=0.135). The results of this study demonstrated that PEMFs were better than TEAS as a non-invasive method to replace the manual acupuncture.

Electrical characterization of proposed transpositional acupoints on the urinary bladder meridian in a rat model.

Specific electrical characteristicsof acupointswere investigated on the urinary bladder (BL) meridian in 14 rats. BL acupointsand non-acupoints on the back were selected and their electrical voltages were measured by using aSPACsystem.The mean voltages of each point or each line were statistically analyzed by using the ANOVA test.The BL meridian showed voltages higher than those of the reference line (P < .05). Bilateral 1st BL lines presented higher voltages than bilateral 2nd BL lines (P < .05). Most BL acupoints had voltageshigher than those for the corresponding reference points (P < .05). In particular, theright BL16 exhibited the biggest difference from the reference point, followed by the left extra BL point-2, the right BL27, the left BL17, and theleft BL45. Additionally, the distributions of neurofilamentsfor several points were investigated by using immunohistochemistry. There was a trend for the BL acupoints to have larger numbers of neurofilaments than the reference points, and that trend seemed to be directly proportional to the difference in voltage between the points.In conclusion, BL acupoints on the back in ratsexhibited specific electric and histologic characteristics. Therefore, those acupointsmay be utilized to investigate the efficacy of acupuncturewith laboratory animals.

Investigation of electrical responses to acupuncture stimulation: the effect of electrical grounding and insulation conditions.

Acupuncture in Oriental medicine has been widely used as a core therapeutic method due to its minimal side-effects and therapeutic efficacy. However, the electrical response to acupuncture stimulation (ERAS) has not been clearly studied under acupuncture conditions that might affect the efficacy of acupuncture therapy. In this study, the ERAS was objectively investigated by measuring meridian electric potentials (MEPs) when the electrical grounding conditions of the operator and subject were varied, and when the insulation conditions of acupuncture needle were varied. MEPs between Sang-geoheo (ST37) and Ha-geoheo (ST39) of the Stomach Meridian (ST) were measured by stimulating Jok-samni (ST36) with an acupuncture needle. For non-insulated acupuncture stimulation (NIAS), the average MEP peak was 148.6 +/- 20.6 when neither the operator nor the subject were electrically grounded, 23.1 +/- 8.8 when the subject only was electrically grounded, 348 +/- 76.8 when the operator only was electrically grounded, and 19.9 +/- 4.7 when both the operator and the subject were electrically grounded. The MEPs presented various magnitudes and patterns depending on the electrical grounding conditions. The MEP pattern was very similar to that of the charge and discharge of a capacitor. For insulated acupuncture stimulation (IAS), the average MEP peak was 20 +/- 4 in all electrical grounding conditions, which is not a significant electric response for acupuncture stimulation. In terms of electricity, this study verified that acupuncture therapy might be affected by acupuncture conditions such as (1) the electrical grounding condition of the operator and the subject and (2) the insulation condition of the acupuncture needle.

Development and clinical testing of an ultrasonic diagnostic system for functional gastrointestinal disorders.

Functional gastrointestinal disorders (FGIDs) affect millions of people without regard for age, race or sex, but little information related to FGIDs and few methods are available for diagnosing FGIDs. Therefore, the aims of this study were to investigate any alterations to the gastrointestinal tracts induced by FGIDs and to develop and test an ultrasonic diagnostic system capable of detecting the alteration in the gastrointestinal tract. Palpation/percussion examinations were performed on 40 participants with or without FGIDs to identify any mechanical alterations to the gastrointestinal tract induced by FGIDs. An ultrasonic diagnostic system for FGIDs was developed and clinical trials were performed on same participants used in the palpation/percussion examinations to test the system under actual clinical conditions. The results obtained from the palpation/percussion examinations showed that a gastrointestinal tract with a FGID became more rigid than a healthy tract (p < 0.05). Clinical trials also showed that gastrointestinal tracts in the patient group with FGIDs were in fact more rigid than those in the control group without FGIDs (p < 0.05). These findings indicated that our newly designed ultrasonic diagnostic system may be useful in clinics to accurately diagnose FGIDs through detecting mechanical alterations to the rigidity of the gastrointestinal tract induced by FGIDs. The current study may be valuable by identifying for the first time mechanical alterations of the gastrointestinal tract induced by FGIDs and may prove valuable as the first step in the development of a diagnostic system for FGIDs.

Development of acupoints discrimination system using electric properties of acupoints and non-acupoints.

The conventional acupuncture-point stimulus pattern uses direct current (DC) to discriminate acupuncture point and meridian lines, which are fundamental to diagnosis and treatment in Oriental medicine. However, DC is insufficiently reliable in discriminating these acupuncture points and lines. To overcome these problems, the authors extracted optimal parameters (e.g., square-like waveform with a frequency of 3 to approximately 5 kHz) with a single power alternating current (SPAC) stimulus pattern of 1.28V. The SPAC stimulus pattern has little or no effect on cells and the body's physiological state and minimizes polarization and ionization of cells because it uses low voltage and alternating current. The authors developed a new meridian discrimination system that can be used with the SPAC.

Effects of acupuncture on potential along meridians of healthy subjects and patients with gastric disease.

The purpose of this study was to investigate the differences in bio-potential in the stomach meridians of 20 normal healthy control subjects (NHC) and 20 patients with gastric disease (PGD). We measured changes in potential between ST-39 and ST-37 on the stomach meridian in response to invasive insertion of a needle at ST-36. The response patterns of NHC were regular in both the left and right meridians, whereas the response patterns of PGD were irregular. The potential of the NHC was significantly higher than that of the PGD group. The potential values in the left and right meridians of NHC were the same. However, the potential in the right meridian was significantly higher than that in the left meridian of PGD (p < 0.001). This study indicates that there may be changes in Qi or in the electrical properties of meridians according to health status, reflected in the regularity of the response to acupuncture or in the level of potential along meridians. Therefore, it may be possible to make a diagnosis from the state of a meridian by comparing levels of potential difference and its regularity.

Differences in electrical conduction properties between meridians and non-meridians.

Therapy using the acupuncture meridian system is an important part of traditional Chinese medicine. The purpose of this study was to investigate the electrical conduction properties of the meridians. The current conduction and potential profiles were compared after switching the current direction in the Hegu (LI-4) and Quchi (LI- 11) meridians and over a non-acupuncture point 1 cm from Quchi (LI-11) in 20 healthy subjects. Both meridians demonstrated significantly higher conductivity between Hegu (LI-4) and Quchi (LI-11) than between Hegu (LI-4) and the non-acupuncture point. The direction of current, peak frequency and absolute potential values in the direction Hegu (LI-4) to Quchi (LI-11) differed significantly from those in the direction Quchi (LI-11) to Hegu (LI-4). These results suggest that the conducting pathways are stronger in the meridians than in the non-meridians and that preferential conduction directions exist between two acupuncture points. These results are consistent with the theories of Qi-circulation and traditional Chinese medicine.

Development of a system and improvement of the stimulus pattern to discriminate the acupuncture point and meridians.

The conventional acupuncture-point stimulus pattern uses direct current (DC) to discriminate acupuncture point and meridian lines, which are fundamental to diagnosis and treatment in Oriental medicine. However, DC is insufficiently reliable in discriminating these acupuncture points and lines. To overcome these problems, the authors extracted optimal parameters (e.g., squarelike waveform with a frequency of 4 kHz) with a single power alternating current (SPAC) stimulus pattern of 1.28 V. The SPAC stimulus pattern has little or no effect on cells and the body's physiological state and minimizes polarization and ionization of cells because it uses low voltage and alternating current. The authors developed a new meridian discrimination system that can be used with the SPAC.

Effects of Qi-therapy (external Qigong) on cardiac autonomic tone: a randomized placebo controlled study.

Heart-rate variability (HRV) was compared in 40 subjects receiving external Qi-therapy (QT) or placebo control therapy, in a randomized placebo controlled design experiment. There were significant time, and group x time interactions in heart rate (HR), low frequency (LF), high frequency (HF), and LF/HF. These suggest that QT reduced the HR and increased HRV as indicated by a reduced LF/HF power ratio of HRV. These findings suggest that QT stabilizes the sympathovagal function more than placebo therapy.

Is there any energy transfer during acupuncture?

Acupuncture therapy is based on the principle of restoring equilibrium of the body's energy state by regulating the flow of Qi in the corresponding meridians and acupoints. The purpose of this study was to determine whether bio-energy transfer occurs during acupuncture of the meridians. We treated 20 normal healthy subjects with acupuncture and measured changes in the direct current (DC) potential between the stomach meridian points ST-39 and ST-37 in response to invasive insertion of a needle at ST-36. Using a crossover study design, the practitioner performed the procedure on each subject once with bare (uncovered) hands (ABH) and once wearing surgical gloves (ASH) to shield the electric energy transfer. ABH produced a response pattern to each stimulation, whereas ASH produced a response pattern only to the first step. ABH elicited a significantly higher response potential than did ASH (p < 0.001). These results suggest that there may be bio-energy (Qi) transfer during acupuncture along the meridians. This transferred bio-energy (Qi) seems to act as an electromotive force, which is purported to remove the stagnation or blockage of energy and restore an equilibrium state.