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American Journal of Chinese Medicine

Evidence of Qi-gong Energy and its Biological Effect on the Enhancement of the Phagocytic Activity of Human Polymorphonuclear Leukocytes

Evidence of Qi-gong Energy and its Biological Effect on the Enhancement of the Phagocytic Activity of Human Polymorphonuclear Leukocytes

Masanori Fukushima

(Accepted for publication December 19, 2000)

Abstract: In order to test for an effect of phosphate buffered saline (PBS) treated externally with Qi energy (“Qi-treated” PBS) on the phagocytic activity of human polymorphonuclear leukocytes (PMNs), rigorously controlled experiments employing masking and randomized procedures were carried out under independent monitoring. In all experiments, Qi treatment was externally applied under monitoring to newly purchased unopened 100 ml bottles of PBS, and the PMN phagocytic activity was assayed by one experimenter in masked, randomized and monitored conditions using a highly sensitive chemiluminescence method. Phagocytic activity data were obtained in triplicate for each sample and then statistically analyzed. The PBS samples Qi-treated by the Qi-gong master and by one of the Qi-gong trainees showed clear stimulation of PMN phagocytic activity which was significant statistically, and this phenomenon was highly reproducible. Out of 10 experiments by the Qi-gong master, only twice did Qi-treatment fail to influence the PBS. The activity of Qi-treated PBS decayed over days or weeks. Furthermore, it was found that Qi-treated PBS had decreased phagocytic stimulatory activity after microwave treatment, but not after autoclave treatment. We also demonstrated that microwave irradiation and infrared laser pulse irradiation have similar effects on PBS as Qi- treatment. The results obtained in this experiment provide evidence of the existence of Qi energy, its ability to influence an electrolyte solution and its biological effect. Furthermore, microwave or infrared laser pulse treatment was found to partly mimic the Qi-treatment of PBS.

Numerous reports have suggested the existence of Qi-gong energy and its biological effects (Sancier et al., 1991; Lu, 1997). Qi-gong currently draws much attention from not only traditional Chinese medicine researchers but also conventional scientists (Ziegler, 1999) and Qi-gong healing is seemingly becoming more accepted as one of the next possible candidates for complementary or alternative medicine, following acupuncture (Sancier et al., 1991). Unfortunately, many reports are described in Chinese or Japanese, and most of these might be considered flawed, since they were undertaken without appropriate masking (blinding) and/or randomization procedures. However, an extensive and systematic review of previous reports allows us certain insights into Qi-gong energy and the application of conventional scientific methods to the study of this subtle energy. Various aspects of Qi-gong energy have been studied by different investigators. These include reports on the electroencephalograms (EEGs) of Qi-gong practitioners (Weixing et al., 1994; Zhang et al., 1988; Liu et al., 1990; Xu et al., 1998) and the effects of Qi-gong on human physiological and pathological conditions, and also the study of cellular activity such as cellular proliferation (Sancier et al., 1991; Lu, 1997; Sancier, 1996; Trieschmann, 1999; Shah et al., 1999; Chien et al., 1991; Wu et al., 1999). Although it is quite difficult to detect Qi-energy using conventional physical measurement techniques, many reports have described specific approaches which detect Qi-gong energy as a magnetic field, far-infrared or photon (Chien et al., 1991; Seto et al., 1992; Hisamitsu et al., 1996; Benford et al., 1999; Machi, 1995; Usa et al., 1995; Lu, 1997). The most notable among them are reports that Qi-gong practitioners emit a strong magnetic field from their hand or head (Hisamitsu et al., 1996; Benford et al., 1999), and the enhancement of natural killer cell-activity in vitro (Yamamoto et al., 1996). Surprisingly, it has even been reported that Qi-gong energy can be stored in various materials (Omura, 1990), despite there being no rational physico-chemical explanation for such an observation at present. At the same time, electromagnetic fields have been found to have a stimulative effect on the phagocytic activity of polymorphonuclear leukocytes (PMNs) (Roy et al., 1995; Gapeyev et al., 1997; Papatheofanis, 1990; Khadir et al., 1999). Microwave or laser irradiation of PMNs has also been shown to enhance their phagocytic activity (Kiel et al., 1986; Dima et al., 1996). Furthermore, recent physico-chemical studies have demonstrated magnetic effects on electrolyte solutions (Oshitani et al., 1999; Oshitani et al., 1999) and their physiological effects (Ayrapetyan et al., 1994), and it has been shown that an aqueous solution pre-exposed to microwave irradiation influences K-Ca channel activity (Fesenko et al., 1995). These reports suggest a “memory effect” in which magnetic effects remain for a considerable period after magnetic exposure is completed (Oshitani et al., 1999; Oshitani et al., 1999; Ayrapetyan et al., 1994; Fesenko et al., 1995; Adair, 1999; Velizarov et al., 1999). Preliminary experiments carried out by Matsumoto and his colleague suggested that external Qi-gong treated phosphate-buffered saline (PBS) has a stimulatory effect on the phagocytic activity of human PMNs assayed by the highly sensitive chemiluminescent method (Kataoka et al., 1997). However, these experiments were not performed with masking and randomized conditions and also large variations in phagocytic activities of PMNs between PMN-donors were not accounted for in the analysis. These several lines of evidence prompted us to carry out a rigorous series of experiments in masked and well-controlled conditions to demonstrate the effect of external Qi-gong treated PBS (referred to as Qi-gongized PBS) on the phagocytic activity of human PMNs using the highly sensitive chemiluminescence assay method.

Materials and Methods

Human Neutrophil Preparation

Three hours before each experiment, human peripheral blood from healthy volunteers was drawn into a tube containing heparin (10 [micro]l/ml blood). PMNs were isolated by centrifugation (400 x g, 60 min) of the blood (4 ml) in a Mono-poly resolving medium (Dai-Nippon Pharmaceutical Co. Ltd. No. 16-980-49) (4 ml) as a single band as described previously (Kataoka et al., 1997; Boyum, 1968; Ting et al., 1971). The isolated PMNs were washed twice with the medium (RPMI- 1640 + 10% FBS GIBCO, No. 11835) by centrifugation (250 x g, 10 min). The PMN preparation was then suspended in the medium at a concentration of 2 x [10.sup.6] cells/ml in a plastic test tube (Falcon No. 35-2095). After a brief gentle vortex, the PMN suspension was carefully aliquotted into the same type of test tube (500 [micro]l/tube) using a pipetter (500 [micro]l tip).

Incubation of Leukocytes with Control PBS or Qi-gongized PBS

500 [micro]l of the control, Qi-gongized or other physical energy-exposed PBS was added to a leukocyte suspension in a test tube prepared as described above. For each PBS sample, triplicate or duplicate incubation was carried out for each PMN donor. After the addition of PBS samples to the PMN suspensions, all tubes were masked and randomly allocated and numbered by the statistician (C.H.) independently in another room. Each of the tubes was inserted into an X-ray-cut (lead) bag (Fuji Film Co. Ltd.) and kept standing separately at a distance of at least 1 m from other tubes and incubated for 1 hr at room temperature. Thus, until the time the assay for PMN phagocytic activity was finished, the tubes which contained Qi-gongized PBS could not be identified except by the statistician (C.H.), and he was not present at the assay of PMN phagocytic activity.

Assay of Phagocytic Activity

After 1 hr incubation at room temperature, all tubes were collected into a tube stand. A triple assay was carried out for each tube using a 96-well micro titer plate (300 [micro]l/well) (Falcon 35-3296) i.e. each of the tubes containing the incubated suspension was gently and carefully pipetted 10 times and 200 [micro]l aliquots of the homogenously dispersed PMN suspension were transferred to the well using a 100 [micro]l tip pipetter. The micro titer plate was then incubated for 10 min under 5% [CO.sub.2] at 37 [degrees] C. After that, luminosphere particles (50 [micro]l, [10.sup.7] particles/well) (Toray Research Center Co. Ltd. LT-5275-2) were added to each well (Aoki et al., 1994; Nishijima et al., 1992; Uchida et al., 1985). In this step, a 12-tip pipetter was used to minimize reaction time variation between wells. The whole procedure usually took less than a minute to complete. Chemiluminescence was then measured at 2 min intervals for 30 min. An ultra sensitive TV camera system (ARGUS-50/MP, Hamamatsu Photonics, K.K.) which can detect luminescence of very small amounts using a 2 dimensional photo counting sensor (C2400-47 HAMAMATSU Photonics K.K.) was used to measure the amount of luminescence. This image processing system fitted with a photon counting TV camera is capable of testing all wells of a 96-well micro titer plate simultaneously (Maly et al., 1988; Suematsu et al., 1988). The use of this system allowed us to assess the phagocytic activity of the PMNs in both the Qi-gongized PBS and those in the control PBS at the same time. When phorborl 12-myristate-13-acetate was diluted to a femto molar level, it was found that this assay method could reliably detect a small enhancement of phagocytosis.

External Qi-gong Treatment of PBS: Profile of Qi-gong Practitioner, Trainees, Screening of People who Have Potential to Emit Qi-gong Energy and Method of Training

Qi-gong master M.M. is 60 and has been practicing Kampo (traditional Chinese herbal) medicine, and so-called Qi-gong healing, for more than 25 years since he opened a Kampo Clinic in 1975. He is a leading acupuncturist and was honored for his efforts to extend the coverage of National Medical Care Insurance to acupuncture therapy in Japan in 1982. He gradually lost his sight after secondary glaucoma due to repeated operations following a traffic accident. He suffered myocardial infarction in 1994 and has diabetes mellitus, hyperglycemia, hyperlipidemia, and impaired renal function. Despite such disabilities, he has treated many patients with Kampo-medicine and Qi-gong every day in his clinic. He has trained several people to emit Qi-gong energy through a training/exercise program that he developed. The training program consists of a controlled breathing method, early morning exercise and the establishment of a very regular life style. Trainee S.M. is a healthy woman of 28 and trainee Y.C. is a healthy woman of 52, who is married with 3 children. Trainee M.K. is a man of 40. The electroencephalogram (EEG) for the Qi-gong master M.M. and the trainee S.M. showed sporadic slow wave (0, 8 wave) activity even when they were alert, whereas the other two trainees (Y.C, M.K.) showed no such slow wave activity during 30 min of EEG recording.

External Qi-gong Treatment of PBS: Procedure and Conditions

Newly purchased, unopened PBS (Dulbecco’s PBS: GiBCO. BRL No. 14040-141) which contains Ca[Cl.sub.2] (anhyd) 0.10, KCl 0.20, [KH.sub.2][PO.sub.4] 0.20, Mg[Cl.sub.2]-6[H.sub.2]O 0.10, NaCl 8.00, [Na.sub.2][HPO.sub.4]-7[H.sub.2]O 2.16; g/l each, (100 ml in plastic bottle) was the subject for external Qi-gong treatment. The PBS bottle cap and wrapping remained unopened until it was used for assaying phagocytic activity. Usually, three PBS bottles were used for the experiment after labeling each bottle with an identification number and masking. Two of the three bottles were then taken by the monitor (M.F.) or statistician (C.H.) to a room in another building for external Qi-gong treatment, which was carried out in a Japanese style tatami-room in the other building, except for the first two experiments (1998.5.7. and 1998.7.5.). The Qi-gong master (M.M.) and/or each trainee (S.M., Y.C. and M.K.) sat down on tatami-mats and concentrated their minds but didn’t meditate or sleep. During this period, the EEG recording showed sporadic [Theta] to [Delta] waves which are not seen during transcendental meditation (Wallace, 1970). Usually at 10 a.m. the practitioners started the Qi-gong treatment procedure. This involved taking a glass test tube (1 cm diameter and 8 cm long, containing a pearl-nucleus which was extensively Qi-treated in advance) in the left hand and an aluminum stick (5 mm diameter, 30 cm long) used for Qi-energy transduction in the right hand. The practitioner pointed to the PBS bottle(s) with the stick without direct touching. During the external Qi-gong emission period, the condition of their mind or consciousness may be important. However, at present, this is only described by their subjective impression based on inner observation. They describe their minds as very silent. This state of mind somewhat differed from meditation. For example, M.M. subjectively interprets this state as unifying his conscious and unconscious minds. Instead of the image of a conscious mind as somehow separate from the unconscious mind that lies below, he senses this Qi-emission state as one where the conscious mind becomes unified with and embedded within a sea of unconsciousness. Throughout the external Qi-gong treatment period, the authors M.F. and C.H. and other monitors T.S. and T.T. carefully watched the practitioners’ condition and behavior. During this period, the practitioners never slept and could talk anytime with the monitors. Therefore, a continuous concentration of their minds was not necessary for Qi-emission. Usually, after two hours of the external Qi-gong treatment, monitors M.F. or C.H. then took the PBS bottle(s) to a preparation room on the same floor of the laboratory in a different building.

Microwave or Laser Treatment

Unopened PBS bottles were exposed to microwaves using a 500W electrical range (NATIONAL NACA5) for 10 sec or 30 sec without rotation. After the PBS temperature cooled down, the PBS was served for phagocytic activity assays. Laser light (670 nm) was used to irradiate the PBS in a quartz cell (NIHON SEKIEI K.K., T-23-ES-10) for 5 min and 10 min using a PLP-02 type pico sec light pulsar (Hamamatsu Photonics K.K.) and then was used in the assay.

Statistical Analysis

First, the phagocytic activity data obtained from each replication was transformed to a logarithmic scale to satisfy the assumption of homogeneity of variance and normal distribution. Two-way (treatment and donor) analysis of variance was performed using the SAS GLM procedure. After confirming that the interaction between donor and treatment was not significant, the main effect of the treatment was tested. If the main effect was significant, the difference (DIF) in the mean between the control and the treatment group was estimated and its 95% confidence interval was calculated using the least square method. Two-tailed statistical significance level was set at 0.05. The enhancement rate was defined as follows.

Enhancement (%) = (exp(DIF) – 1) x 100 = (XT/XC – 1) x 100

DIF: the difference in the mean between the control and the treatment in logarithmic scale.

XT: geometric mean in the treatment group;

XC: geometric mean in the control group.

Results

From May 7, 1998 to December 29, 1999, 16 experimental sessions were carried out. One Qi-gong practitioner and 2 trainees performed external Qi-gong treatment of PBS. In all experiments, PMN phagocytic activity was assayed by one experimenter (T.K.) blindly under rigorously controlled conditions as described in the Methods section. The phagocytic activity of PMNs has a large variation among donors and even within an individual (see Table 3: Experiment series 1 and 2). From the 3rd experiment in experimental series 1 we established the following procedure: in order to deliver an equal number of PMNs into each tube by pipetting and to ensure this number was maintained without loss by adhesion, the pipetting was standardized and the tubes were carefully maintained in a vertical position. The whole experimental process was carefully monitored by two of the authors (M.F. and C.H.) and other third parties (T.T. and T.S.). All assay procedures were finished within one hour. The Coefficients of Variation (CV) of phagocytic activity in control groups assayed through the whole experimental series fell into a range from 0 to 44% (average of CV was 11%). As actual examples of experiments, Figure 1 shows the luminescence of 96 wells 15 min after the addition of lumispheres at 37 [degrees] C for the PBS samples from experimental series 2 (1999.10.23 and 1999.11.21) by Qi-gong trainees S.M and M.K.. Table 1 records the actual amount of luminescence measured by an ultra sensitive TV camera system as described in Materials and Methods and each number is matched to the sample blinded before the assay. Table 2 summarizes the calculation of mean values and SDs for PMN phagocytic activity expressed as luminescence and shows the statistical analysis of these results as described in Methods. The PBS treated by the Qi-gong trainee S.M. significantly enhanced phagocytic activity compared to controls, whereas the PBS treated by Qi-gong trainee M.K. did not show enhancement. Table 3 summarizes the results of the whole experimental series. PBS treated by Qi-gong master M.M. and Qi-gong trainee S.M. showed potent stimulation of PMN phagocytic activity and this phenomenon was highly reproducible while the PBS treated by trainee Y.C. showed such stimulation only once (1999.5.23). Out of 10 experiments by Qi-gong master M.M. only twice (1st: 1998.5.7; 9th: 1999.8.8.) did the PBS not show enhancement. In the 2nd experiment (1998.7.5.) we found that the p value showed statistical significance for sample No. 2 when the phagocytic activity was adjusted for actual cell number. Thereafter we standardized the pippetting and handling procedure for PMNs to minimize the deviation of delivered cell numbers. Furthermore, from the 3rd experiment onwards, the treatment by the Qi-gong practitioner was carried out in an isolated calm clean tatami-room as described in the Methods. During the 3rd experiment negative energy was accidentally produced because M.M. gripped negative Qi-treated material with the left hand by mistake. Thereafter the assay was very stable and the highly positive results were reproducible. During the period when the 8th and 9th experiments were carried out, the Qi-gong master’s health condition was very bad since he had four teeth extracted at once by a dentist on May 20, 1999. After the extraction of his teeth, he had taken anti-inflammatory drugs and antibiotics for several months. He needed a number of months for his recovery. It should be noted that while his physical condition was bad, the degree of his Qi-gongization was very poor, and no significant effect was observed on 1999.8.8. The time course of Qi-gongized PBS activity for the enhancement of PMN phagocytosis was followed at one week intervals for the sample prepared by M.M. on November 8, 1998, and at 3 day intervals for the sample treated by S.M. on Nov. 21, 1999. The activity of the samples decayed over weeks or days respectively (Table 4). It was found that the Qi-gongized PBS (M.M. sample 1998.11.21., 1999.4.3., 1999.4.11.) shows decreased phagocytic stimulatory activity after microwave treatment (electric range, 20 sec, 500 W with rotation), although not after autoclave treatment (120 [degrees], 1.5 a.p., 20 min) (Table 5). Finally, we examined PBS exposed to microwaves from the electric range without rotation and a pulse laser at 670 nm. As shown in Table 6, such physical treatment of PBS was associated with significant enhancement of PMN phagocytic activity. However, this activity diminished within several hours at room temperature.

[Figure 1 ILLUSTRATION OMITTED]

Table 1A. Raw Data (Y) of Phagocytic Activity for Each PBS Sample for

PMNs Taken from 5 Donors. The Amount of Luminescence Shown in Figure 1

was Represented as a Digital Value Using the Two Dimensional

Photocounting Sensor (C2400-47 HAMAMATSU Photonics K.K.) Described in

Materials and Methods

OBS DONOR GROUP REP ID I Y

1 a 3miya 1 1 1 71.3

2 a 3miya 1 1 2 75.4

3 a 3miya 1 1 3 117.8

4 a 2kubo 2 2 1 58.8

5 a 2kubo 2 2 2 55.8

6 a 2kubo 2 2 3 61.4

7 a 3miya 2 3 1 83.5

8 a 3miya 2 3 2 64.7

9 a 3miya 2 3 3 69.3

10 a 1control 1 4 1 62.6

11 a 1control 1 4 2 58.3

12 a 1control 1 4 3 62.2

13 a 1control 2 5 1 69.3

14 a 1control 2 5 2 57.9

15 a 1control 2 5 3 52.2

16 a 2kubo 1 6 1 45.3

17 a 2kubo 1 6 2 37.9

18 a 2kubo 1 6 3 52.2

19 b 1control 1 7 1 15.5

20 b 1control 1 7 2 15.1

21 b 1control 1 7 3 13.0

22 b 2kubo 1 8 1 12.0

23 b 2kubo 1 8 2 9.7

24 b 2kubo 1 8 3 9.2

25 b 3miya 1 9 1 39.2

26 b 3miya 1 9 2 34.4

27 b 3miya 1 9 3 35.7

28 b 2kubo 2 10 1 12.3

29 b 2kubo 2 10 2 10.6

30 b 2kubo 2 10 3 10.7

31 b 3miya 2 11 1 27.6

32 b 3miya 2 11 2 25.5

33 b 3miya 2 11 3 32.5

34 b 1control 2 12 1 18.6

35 b 1control 2 12 2 17.8

36 b 1control 2 12 3 20.8

37 c 1control 1 13 1 11.0

38 c 1control 1 13 2 7.6

39 c 1control 1 13 3 9.4

40 c 2kubo 1 14 1 7.7

41 c 2kubo 1 14 2 6.4

42 c 2kubo 1 14 3 10.6

43 c 2kubo 2 15 1 10.4

44 c 2kubo 2 15 2 10.3

45 c 2kubo 2 15 3 10.1

46 c 3miya 1 16 1 58.2

47 c 3miya 1 16 2 49.9

48 c 3miya 1 16 3 49.6

49 c 3miya 2 17 1 55.1

50 c 3miya 2 17 2 58.8

51 c 3miya 2 17 3 56.0

52 c 1control 2 18 1 15.2

53 c 1control 2 18 2 9.3

54 c 1control 2 18 3 8.9

55 d 2kubo 1 19 1 10.7

56 d 2kubo 1 19 2 10.0

57 d 2kubo 1 19 3 9.0

58 d 3miya 1 20 1 41.9

59 d 3miya 1 20 2 46.2

60 d 3miya 1 20 3 39.6

61 d 2kubo 2 21 1 11.7

62 d 2kubo 2 21 2 12.6

63 d 2kubo 2 21 3 13.6

64 d 1control 1 22 1 12.0

65 d 1control 1 22 2 10.0

66 d 1control 1 22 3 11.7

67 d 1control 2 23 1 15.8

68 d 1control 2 23 2 13.8

69 d 1control 2 23 3 11.8

70 d 3miya 2 24 1 36.9

71 d 3miya 2 24 2 37.6

72 d 3miya 2 24 3 33.3

73 e 3miya 1 25 1 36.9

74 e 3miya 1 25 2 41.1

75 e 3miya 1 25 3 35.6

76 e 1control 1 26 1 13.8

77 e 1control 1 26 2 10.1

78 e 1control 1 26 3 11.8

79 e 1control 2 27 1 11.6

80 e 1control 2 27 2 10.4

81 e 1control 2 27 3 13.0

82 e 3miya 2 28 1 34.4

83 e 3miya 2 28 2 38.9

84 e 3miya 2 28 3 32.3

85 e 2kubo 1 29 1 6.9

86 e 2kubo 1 29 2 6.7

87 e 2kubo 1 29 3 7.4

88 e 2kubo 2 30 1 5.7

89 e 2kubo 2 30 2 6.3

90 e 2kubo 2 30 3 6.0

External Qi-gong treated PBS by Qi-gong trainees S.M. (miya)

and M.K. (kubo) on experiment date, 1999.10.23.

REP: replication of Group I: replication of well

Table 1B. Raw Data (Y) of Phagocytic Activity for Each PBS Sample

for PMNs Taken from 5 Donors. The Amount of Luminescence Shown in

Figure 1 was Represented as a Digital Value Using the Two

Dimensional Photocounting Sensor (C2400-47 HAMAMATSU Photonics

K.K.) Described in Materials and Methods

OBS DONOR GROUP REP ID I Y

1 a 1miya 1 1 1 355.0

2 a 1miya 1 1 2 325.7

3 a 1miya 1 1 3 392.2

4 a 1miya 2 2 1 335.0

5 a 1miya 2 2 2 207.0

6 a 1miya 2 2 3 315.2

7 a 3control 1 3 1 72.7

8 a 3control 1 3 2 93.4

9 a 3control 1 3 3 102.7

10 a 2control 1 4 1 136.1

11 a 2control 1 4 2 116.5

12 a 2control 1 4 3 130.8

13 a 3control 2 5 1 107.8

14 a 3control 2 5 2 104.8

15 a 3control 2 5 3 88.8

16 a 2control 2 6 1 94.3

17 a 2control 2 6 2 110.8

18 a 2control 2 6 3 109.7

19 b 2control 1 7 1 63.8

20 b 2control 1 7 2 75.1

21 b 2control 1 7 3 49.0

22 b 1miya 1 8 1 490.5

23 b 1miya 1 8 2 489.8

24 b 1miya 1 8 3 396.2

25 b 3control 1 9 1 55.8

26 b 3control 1 9 2 72.8

27 b 3control 1 9 3 78.6

28 b 1miya 2 10 1 487.5

29 b 1miya 2 10 2 575.5

30 b 1miya 2 10 3 453.1

31 b 2control 2 11 1 61.3

32 b 2control 2 11 2 47.0

33 b 2control 2 11 3 63.9

34 b 3control 2 12 1 67.8

35 b 3control 2 12 2 74.5

36 b 3control 2 12 3 71.8

37 c 1miya 1 13 1 729.4

38 c 1miya 1 13 2 558.4

39 c 1miya 1 13 3 465.9

40 c 3control 1 14 1 116.7

41 c 3control 1 14 2 137.5

42 c 3control 1 14 3 185.7

43 c 2control 1 15 1 193.3

44 c 2control 1 15 2 201.8

45 c 2control 1 15 3 236.4

46 c 2control 2 16 1 142.0

47 c 2control 2 16 2 119.0

48 c 2control 2 16 3 90.1

49 c 1miya 2 17 1 695.4

50 c 1miya 2 17 2 740.7

51 c 1miya 2 17 3 841.3

52 c 3control 2 18 1 189.7

53 c 3control 2 18 2 194.5

54 c 3control 2 18 3 167.8

55 d 1miya 1 19 1 1478.3

56 d 1miya 1 19 2 1840.6

57 d 1miya 1 19 3 1641.4

58 d 2control 1 20 1 288.9

59 d 2control 1 20 2 304.2

60 d 2control 1 20 3 362.3

61 d 1miya 2 21 1 2010.2

62 d 1miya 2 21 2 1801.6

63 d 1miya 2 21 3 1707.7

64 d 3control 1 22 1 227.1

65 d 3control 1 22 2 225.6

66 d 3control 1 22 3 280.8

67 d 3control 2 23 1 282.9

68 d 3control 2 23 2 322.7

69 d 3control 2 23 3 305.9

70 d 2control 2 24 1 371.8

71 d 2control 2 24 2 293.2

72 d 2control 2 24 3 277.0

73 e 2control 1 25 1 35.6

74 e 2control 1 25 2 59.4

75 e 2control 1 25 3 68.2

76 e 3control 1 26 1 57.6

77 e 3control 1 26 2 60.5

78 e 3control 1 26 3 51.1

79 e 2control 2 27 1 56.7

80 e 2control 2 27 2 45.1

81 e 2control 2 27 3 57.5

82 e 3control 2 28 1 55.1

83 e 3control 2 28 2 47.1

84 e 3control 2 28 3 48.1

85 e 1miya 1 29 1 241.4

86 e 1miya 1 29 2 242.3

87 e 1miya 1 29 3 188.5

88 e 1miya 2 30 1 247.7

89 e 1miya 2 30 2 239.6

90 e 1miya 2 30 3 265.7

External Qi-gong treated PBS by Qi-gong trainee S.M. (miya) on

experiment date, 1999.11.21.

REP: replication of Group I: replication of well

Table 2. Representation of Phagocytic Activities in 5 Donors’ PMNs

with the Mean Value Calculated from the Raw Data Presented in Table 1

and SD for Each PBS Sample

PMN donor PBS Sample N Obs Mean SD Minimum Maximum

Experiment date 1999.10.23.

age/sex

a. 24 F 1 control 2 60.4 0.9 59.8 61.1

2 M.K. 2 51.9 9.6 45.1 58.7

3 S.M. 2 80.3 11.1 72.5 88.2

b. 44 F 1 control 2 16.8 3.2 14.5 19.1

2 M.K. 2 10.8 0.6 10.3 11.2

3 S.M. 2 32.5 5.6 28.5 36.4

c. 45 F 1 control 2 10.2 1.3 9.3 11.1

2 M.K. 2 9.3 1.4 8.2 10.3

3 S.M. 2 54.6 2.9 52.6 56.6

d. 50 F 1 control 2 12.5 1.8 11.2 13.8

2 M.K. 2 11.3 1.9 9.9 12.6

3 S.M. 2 39.3 4.7 35.9 42.6

e. 37 F 1 control 2 11.8 0.2 11.7 11.9

2 M.K. 2 6.5 0.7 6.0 7.0

3 S.M. 2 36.5 1.9 35.2 37.9

Experiment date 1999.11.21.

a. 50 M 1 S.M. 2 321.7 50.8 285.7 357.6

2 control 2 116.4 16.2 104.9 127.8

3 control 2 95.0 7.6 89.6 100.4

b. 50 F 1 S.M. 2 482.1 32.9 458.8 505.4

2 control 2 60.0 3.7 57.4 62.6

3 control 2 70.2 1.6 69.1 71.4

c. 54 M 1 S.M. 2 671.9 123.4 584.6 759.1

2 control 2 163.8 66.1 117 210.5

3 control 2 165.3 26.4 146.6 184.0

d. 44 F 1 S.M. 2 1746.6 131.8 1653.4 1839.8

2 control 2 316.2 3.2 314 318.5

3 control 2 274.2 42 244.5 303.8

e. 16 M 1 S.M. 2 237.5 19 224.1 251.0

2 control 2 53.8 0.9 53.1 54.4

3 control 2 53.3 4.5 50.1 56.4

The initials indicate Qi-gong trainees; Triple assays per observation;

Minimum and maximum represent the averages of triple assays for two

observations. Statistical Analysis as follows:

Group vs. Group Enhancement 95% CI P value

Experiment date 1999.10.23.

1 control 1 M.K. -24.531 -42.253 -1.371 0.040

1 control 2 S.M. 166.131 103.637 247.804 0.000

2 M.K. 3 S.M. 252.638 169.83 360.88 0.000

Experiment date 1999.11.21.

1 control 2 control -3.072 -20.412 18.045 0.746

2 control 3 S.M. 389.853 285.8 472.217 0.000

2 control 3 S.M. 384.746 298.029 490.355 0.000

Definition of enhancement of the phagocytic activity in each sample

and the statistical analysis are described in the statistical

analysis section.

Table 3. Effect of Qi-gong Treated PBS on Human PMN Phagocytosis,

Summary of the Results of All Experiments

Date PBS sample(s) PMN donor(a/)

Experiment Series 1 (M.M.)

n

1. 1998 5. 7 1 2

2. 1998. 7. 5 2 (No. 1) 2

(No. 2)

* Adjusted to (No. 1)

cell count (No. 2)

3. 1998. 9. 27(c/) 2 (No. 1) 3

(No. 2) 3

4. 1998. 11. 8 2 (No. 1) 3

(No. 2) 3

5. 1998. 12. 27 2 (No. 1) 2

(No. 2) 2

6. 1999. 3. 21 1 5

7. 1999. 5. 23 2 (No. 1) 3

(No. 2)(d/) 3

8. 1999. 6. 13(e/) 2 (No. 1) 3

(No. 2) 3

9. 1999. 8. 8(e/) 1 (No. 1) 3

(No. 2) 3

10. 1999. 12. 29 2 (No. 1) 3

(No. 2)(d/) 3

Experiment Series 2 (S.M.)

1. 1999. 3. 21 1 5

2. 1999. 4. 18 1 4

3. 1999. 5. 23 1 4

4. 1999. 6. 13 1 5

5. 1999. 10. 11 2 (No. 1) 5

(No. 2) 5

6. 1999. 10. 23 1 5

7. 1999. 11. 7 1 5

8. 1999. 11. 21 1 5

Experiment Series 3 (Y.C.)

1. 1999. 4. 18 1 4

2. 1999. 5. 23 1 4

3. 1999. 6. 13 1 5

4. 1999. 11. 7 1 5

Date PBS sample(s) Enhancement

Experiment Series 1 (M.M.)

%

1. 1998 5. 7 1 57

2. 1998. 7. 5 2 (No. 1) 13

(No. 2) 220

* Adjusted to (No. 1) 23

cell count (No. 2) 438

3. 1998. 9. 27(c/) 2 (No. 1) -69

(No. 2) -72

4. 1998. 11. 8 2 (No. 1) 273

(No. 2) 219

5. 1998. 12. 27 2 (No. 1) 64

(No. 2) 78

6. 1999. 3. 21 1 144

7. 1999. 5. 23 2 (No. 1) 104

(No. 2)(d/) 138

8. 1999. 6. 13(e/) 2 (No. 1) -8

(No. 2) -10

9. 1999. 8. 8(e/) 1 (No. 1) -11

(No. 2) -9

10. 1999. 12. 29 2 (No. 1) 16

(No. 2)(d/) 22

Experiment Series 2 (S.M.)

1. 1999. 3. 21 1 174

2. 1999. 4. 18 1 -3

3. 1999. 5. 23 1 35

4. 1999. 6. 13 1 39

5. 1999. 10. 11 2 (No. 1) 67

(No. 2) -9

6. 1999. 10. 23 1 166

7. 1999. 11. 7 1 24

8. 1999. 11. 21 1 385

Experiment Series 3 (Y.C.)

1. 1999. 4. 18 1 -1

2. 1999. 5. 23 1 198

3. 1999. 6. 13 1 8

4. 1999. 11. 7 1 9

Date PBS sample(s) P value(b/)

Experiment Series 1 (M.M.)

1. 1998 5. 7 1 0.311

2. 1998. 7. 5 2 (No. 1) 0.829

(No. 2) 0.069

* Adjusted to (No. 1) 0.514

cell count (No. 2) 0.001

3. 1998. 9. 27(c/) 2 (No. 1) 0.000

(No. 2) 0.000

4. 1998. 11. 8 2 (No. 1) 0.000

(No. 2) 0.000

5. 1998. 12. 27 2 (No. 1) 0.009

(No. 2) 0.004

6. 1999. 3. 21 1 0.000

7. 1999. 5. 23 2 (No. 1) 0.002

(No. 2)(d/) 0.000

8. 1999. 6. 13(e/) 2 (No. 1) 0.015

(No. 2) 0.002

9. 1999. 8. 8(e/) 1 (No. 1) 0.158

(No. 2) 0.280

10. 1999. 12. 29 2 (No. 1) 0.026

(No. 2)(d/) 0.004

Experiment Series 2 (S.M.)

1. 1999. 3. 21 1 0.000

2. 1999. 4. 18 1 0.737

3. 1999. 5. 23 1 0.012

4. 1999. 6. 13 1 0.000

5. 1999. 10. 11 2 (No. 1) 0.009

(No. 2) 0.592

6. 1999. 10. 23 1 0.000

7. 1999. 11. 7 1 0.009

8. 1999. 11. 21 1 0.000

Experiment Series 3 (Y.C.)

1. 1999. 4. 18 1 0.946

2. 1999. 5. 23 1 0.000

3. 1999. 6. 13 1 0.337

4. 1999. 11. 7 1 0.280

In parenthesis the initial indicate Qi-gong master (M.M.) and trainees

(S.M. & Y.C.).

(a/): N=Number of PMN donors. Assay for each donor’s PMN was

duplicated.

(b/): Definition of enhancement of the phagocytic activity in each

sample and the statistical analysis are described in the statistical

analysis section.

(c/): In this experiment, accidentally the Qi-gong master treated the

PBS while gripping the “negative energy standard” in his left hand.

(d/): In these experiments, during the Qi-treatment bottle No. 2 was

shielded in X-ray-cut (lead) bag, and bottle No. 1 was wrapped in a

paper bag as sham shield, i.e. control for lead bag.

(e/): In this experiment the assay was carried out 3 days after

the Qi-gongization.

Table 4. Decay of Phagocytosis-enhancement Effect of Qi-treated PBS

Date Period PMN donor Enhancement(a/)

Experiment Series 1 (M.M.)

weeks n %

1. 1998 11. 8 — 3 273

1. 1998. 11. 15 1 2 580

1. 1998. 11. 21 2 2 360

1. 1998. 11. 28 3 2 263

1. 1998. 12. 3 4 2 63

1. 1998. 12. 10 5 2 51

1. 1998. 12. 17 6 2 44

Experiment: Series 2 (S.M.)

days n %

1. 1998. 11. 21 — 3 385

1. 1998. 11. 24 3 3 104

1. 1998. 11. 27 6 3 4

Date Period 95% CI P value(a/)

Experiment Series 1 (M.M.)

weeks Lower Upper

1. 1998 11. 8 — 134 496 0.000

1. 1998. 11. 15 1 430 773 0.000

1. 1998. 11. 21 2 238 526 0.000

1. 1998. 11. 28 3 165 397 0.000

1. 1998. 12. 3 4 29 107 0.100

1. 1998. 12. 10 5 9 111 0.025

1. 1998. 12. 17 6 17 78 0.003

Experiment: Series 2 (S.M.)

days Lower Upper

1. 1998. 11. 21 — 298 490 0.000

1. 1998. 11. 24 3 64 152 0.000

1. 1998. 11. 27 6 -6 15 0.447

(a/): Defined as described in the legend for Table 3.

Table 5. Effect of Microwave (MW) Treatment on Qi-treated PBS

in Relation to Phagocytic Enhancement of PMN

Treatment Phagocytic Activity(a/)

Date Sample PMN donor Control PBS

Microwave Qi-treatment

date Age/Sex

1998.11.21. 11.8.(No. 2) 36 M 25.3

33 M 60.2

40 F 57.2

1999. 4. 3. 3.21. 36 M 123.2

27 M 51.2

38 M 103.2

20 M 95.3

1999. 4.11. 3.21. 36 M 103.2

27 M 85.8

38 M 138.2

20 M 78.4

Autoclave

1998.11.21. 11.8 (No. 2) 18 M 27.3

19 M 84.2

Treatment Phagocytic Activity(a/)

Date Sample PMN donor Qi-treated PBS

Microwave Qi-treatment MW Treatment

date Age/Sex Before After

1998.11.21. 11.8.(No. 2) 36 M 45.2 (79) 38.0 (50)

33 M 90.2 (50) 70.5 (17)

40 F 68.3 (19) 63.9 (11)

1999. 4. 3. 3.21. 36 M 195.3 (59) 132.7 (29)

27 M 94.2 (84) 92.0 (80)

38 M 170.3 (65) 128.2 (24)

20 M 200.3 (110) 139.5 (46)

1999. 4.11. 3.21. 36 M 148.3 (44) 132.7 (29)

27 M 127.3 (48) 116.3 (36)

38 M 183.2 (33) 152.2 (10)

20 M 122.4 (56) 139.5 (78)

Autoclave

1998.11.21. 11.8 (No. 2) 18 M 157.3 (476) 136.3 (399)

19 M 245.3 (191) 239.0 (183)

Treatment Phagocytic Activity(a/)

Date Sample PMN donor

Microwave Qi-treatment

date Age/Sex % decrease

1998.11.21. 11.8.(No. 2) 36 M 37

33 M 66

40 F 42

1999. 4. 3. 3.21. 36 M 86

27 M 5

38 M 63

20 M 58

1999. 4.11. 3.21. 36 M 34

27 M 25

38 M 70

20 M -39

Autoclave

1998.11.21. 11.8 (No. 2) 18 M 16

19 M 4

In parentheses: % increase of the phagocytic activity to the control

(a/): single assay for each donor’s PMN, each value represents the

average of 3 wells as described in the Methods.

Table 6. Effect of PBS Exposed to Various Types of Physical Energy

on Phagocytic Activity of Human PMN

PMN donor Treatmer Treatment N Mean SD

age/sex

1 50 M 1 control 3 13.9 0.5

2 microwave 10 sec 3 28.3 1.7

3 microwave 30 sec 3 24.4 2.5

4 laser 5 min 3 21 2

5 laser 10 mihn 3 20.7 2.5

2 50 F 1 control 3 7.6 2.7

2 microwave 10 sec 3 8.6 0.4

3 microwave 30 sec 3 9.8 0.4

4 laser 5 min 3 8.5 0.4

5 laser 10 min 3 9.8 1.5

PMN donor Treatmer Treatment Minimum Maximum

age/sex

1 50 M 1 control 13.3 14.2

2 microwave 10 sec 26.6 29.9

3 microwave 30 sec 21.6 26.2

4 laser 5 min 19.2 23.2

5 laser 10 mihn 18.2 23.1

2 50 F 1 control 4.5 9.4

2 microwave 10 sec 8.3 9.1

3 microwave 30 sec 9.5 10.3

4 laser 5 min 8.2 9

5 laser 10 min 8.3 11.3

Statistical Analysis

Group vs Group Enhancement 95% CI

1 microwave 10 sec 1 control 55.207 29.037 86.683

2 microwave 30 sec 1 control 53.983 28.020 85.212

3 laser 5 min 1 control 33.195 10.737 60.207

4 laser 10 min 1 control 40.873 17.121 69.443

Group P value

1 microwave 10 sec 0.000

2 microwave 30 sec 0.000

3 laser 5 min 0.020

4 laser 10 min 0.004

670 nm laser

Discussion

This rigorous experimental series has demonstrated that PBS treated with external Qi-gong, by not only the Qi-gong master but also one of the Qi-gong trainees, showed markedly significant stimulation of the phagocytic activity of PMNs as assayed by the highly sensitive chemiluminescence method. The authors believe this report provides rigorous objective evidence of the existence of Qi-gong energy, and some of its biological effects. Surprisingly, it was demonstrated that Qi-gong energy might be stored in PBS as suggested by Omura (Omura, 1990); alternatively it could change the physico-chemical nature of PBS (Adair, 1999). Our observations raise the following issues: 1. Qi-gong practitioners or a specifically trained person emits energy X. 2. Energy X can be stored in PBS or energy X changes the physico-chemical nature of the PBS. 3. Qi-gong treated PBS enhances the phagocytic/bactericidal activity of human PMNs. Several reports have suggested that Qi-gong practitioners emit strong magnetic fields and the same research group reported that this magnetic field emitted by particular people stimulates human NK activity (Yamamoto et al., 1996). We have demonstrated that microwave irradiation and laser pulse irradiation also show a very similar effect on PBS as Qi-gong energy in that microwave or pulse laser treated PBS showed significant enhancement of PMN phagocytosis. This phenomenon might have some relation to the magnetic effect on aqueous solutions reported by Higashitani’s group and others (Oshitani et al., 1999; Oshitani et al., 1999; Ayrapetyan et al., 1994; Fesenko et al., 1995; Adair, 1999; Velizarov, 1999). At present no existing physical theory can explain this memory effect and it is not clear whether Qi-gong energy is fully explained by the magnetic field itself. One possible explanation for the nature of energy X is that it is an intensified, pulsed bioelectromagnetic field, and this is generated and emitted in a particular conscious state. If so, the question is where is the site of energy generation and how is it generated. Furthermore, what is the physiological significance for human beings and is such ability specific to human beings. There are other possibilities, for example, the electromagnetic energy field in space could be gathered, transduced and amplified through the human body and put out as a pulsed electromagnetic field. However, we cannot exclude the possibility that Qi-gong energy is in fact an unknown form of energy that is not an electromagnetic field (Ziegler, 1999). One mysterious phenomenon is the negative effect of Qi-gong energy, which was observed twice with M.M. (1998.9.27; 1999.6.13). This may support the reports that there are two types of Qi-gong energy: positive and negative (Sancier et al., 1991; Chien et al., 1991; Wu et al., 1999; Omura, 1990). In the case of electromagnetic radiation, Gapeyev et al. (Gapeyev et al., 1997) observed that the respiratory bursts of neutrophils could be activated or inhibited by an electromagnetic field as a function of modulation frequency. This observation might provide some hints for further studies on this dual effect of Qi-gong energy. The nature of Qi-gong energy should be identified and characterized in the future and the conscious state, which is essential for Qi-gong energy emission, should also be the subject of rigorous objective study using sophisticated diagnostic technologies such as EEG, magnetoencephalogram, magnetic resonance imaging, single-photon emission CT, and positron emission tomography. At present, the only evidence we have is the EEG pattern obtained from M.M. and S.M. The Qi-gong consciousness state is quite different from others in terms of the recording of sporadic slow waves ranging from [Delta] to [Theta] which usually appear in the induction state of sleeping. Such observations are consistent with the report by Weixing et al. (Weixing et al., 1994). However, whether or how this conscious state relates to Qi-gong energy emission remains unclear. Major points, which should be emphasized to reproduce the phenomena observed in our experiments, are: 1. Selection of Qi-gong energy emitter, 2. Setting up adequate conditions for Qi-gong emission, 3. A sensitive and stable assay method, 4. Adequate statistical design and analysis of all data. Our observations demonstrate that Qi-gong energy is either stored as such in PBS or promotes the formation of a very stable molecular species in PBS. At present, in fact, the number of people who emit stable and strong Qi-gong energy is very limited. However, the findings described in this paper warrant further careful extensive studies on Qi-gong energy itself, and its applications not only in a clinical setting but also in other beneficial fields for human life. Furthermore, studies on the Qi-gong consciousness state may be a very promising avenue to understanding and controlling the structure and functions of consciousness.

Acknowledgments

The authors are indebted to the Qi-gong trainees: Ms Sayuri Miyazaki, Ms Yoshiko Chihara and Mr. Mamoru Kuboyama for their very kind cooperation during the study and also to Ms Yurie Kanazawa at Jo-yuin Laboratories for her excellent assistance to Mippo Matsumoto (M.M.) and to many PMN donors. We are grateful to Professor Tai Takahashi at the Kokusai Health and Welfare College for his initial support for M.M.’s desire to scientifically prove the existence of Qi-gong energy which led to working with T.K. in the earlier laboratory experiments and also for his cooperation in monitoring the experiments with Mr. Tsuyoshi Shirai at the Shionogi Pharmaceutical Co. Ltd., and Professor Gen Sobue at the Nagoya University School of Medicine, Department of Neurology for his diagnosis of EEG of the Qi-gong practitioner and trainees, and to Ms Mariko Goan for her excellent secretarial assistance.

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Masanori Fukushima(1)(*), Takuji Kataoka(2), Chikuma Hamada(1) and Mippo Matsumoto(3) (1) Department of Pharmacoepidemiology, School of Public Health, Kyoto University, Yoshidakonoe-cho, Sakyo-ku, Kyoto 606-8501, Japan (2) System Division, Hamamatsu Photonics, Co. Ltd. 325-6 Sunayama-cho, Hamamatsu, 430-8587, Japan and (3) Jo-yuin RBP Laboratories, 645-12 MinamiKoriyama-cho, YamatoKoriyama, Nara, 639-1007, Japan (*) Corresponding author

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