靈氣能治療癌細胞

Reiki can treat cancer cells 靈氣能治療癌細胞 
Nadejda Rozanova University of California, Santa Cruz 

INTRODUCTION 導論 
The number of people who seek Complementary Alternative Medicine (CAM) as a way to address their health care needs has increased significantly over the last several decades. In the future, CAM is expected to continue playing an augmented role in medical practice, including the treatment of cancer [1]. One area of CAM is a biofield medicine that involves subtle body energies, often called Ki or Chi. There exist a substantial number of basic scientific studies on biofield modalities for cancer treatment, such as Qigong, Johrei, Nishino Ki healing, and spiritual healing [2-6]. One study [5] showed no effect of spiritual healing on in vitro tumor cell proliferation. A contrasting study [7] showed that energy healing such as Therapeutic Touch appears to decrease differentiation and mineralization in a human osteosarcoma-derived cell line after two weeks of treatment. 
尋求輔助替代療法(CAM)作為解決其醫療保健需求的一個方法的人數在過去幾十年中大大增加。 在未來,預計 CAM 將繼續在醫療實踐中發揮更大的作用,包括治療癌症。輔助替代療法的領域是生 物醫學,涉及微妙的身體能量,通常稱之為 Ki or Chi。 對於癌症治療的生物場模式存在大量的基礎科學研究,如氣功,上光靈氣,西野靈氣療癒以及靈修。 一項研究顯示,精神治療對玻璃內腫瘤細胞增殖沒有影響。一個對比的研究顯示,能量療癒例如治 療觸摸治療兩週後,可減少在人骨肉瘤來源細胞的分化和礦化。 

We decided to test the effect of energy healing on cancer cells during in vitro experiments, using a similar protocol to that used in the Zachariae study. The aim of this study was to test the hypothesis that spiritual healing will reduce proliferation and viability of two cancer cell lines in vitro. Three controlled experiments were conducted with three different healers and randomized allocation of cells to five different doses of healing or control. Main outcome measures were MTT viability, 3H-thymidine incorporation and counts of an adherent human breast cancer cell line (MCF-7), and a non-adherent mouse B-lymphoid cell line (HB-94). In this study Analyses of variance (ANOVAs) revealed no significant main or dose-related effects of spiritual healing compared to controls for either of the two cell lines or any of the assays (P-values between 0.09 and 0.96). [5]. 
我們決定在玻璃內實驗中使用與 Zachariae 研究中使用的相似的方案來測試能量療癒對癌細胞的 作用。這項研究的目的是為檢測靈療將減少兩種癌細胞株玻璃內增殖和生存能力這一假設。3 位不 同療癒者和隨機分配 5 種不同療癒或控制劑量進行 3 種對照。主要指標是 MTT 生存能力,氚嵌入和 計算人類乳癌細胞(MCF-7)以及非附著老鼠 B 淋巴球細胞(HB-94)。在這個研究中顯示靈修劑量相關 因素與對照兩種細胞株或任何測定法的對照相比,沒有顯著意義(P 值 0.09-0.96)。 

We hypothesized that the problem with this study was its short length. And we decided to use the prolonged study, postulating that the failure of some in vitro studies to show positive effect depends on their longevity and on the number of healing interventions. For example, the effect of energy healing on cells was demonstrated only after a prolonged period in a study conducted by Jhaveri [7]. In our experiments, we used biofield healers trained in the Reiki technique. Reiki is a biofield medicine originally from Japan, now used throughout the world [8-11]. We used an MCF-7 cell line and three Reiki-trained healers. Healers were Reiki Master (2 years experienced) and two other students with Reiki second degree (1 year experienced). There were untreated control and “placebo healing” with an inexperienced person performing the treatment. We used the cell counting method, which includes direct cell counting with Coulter Counter model Z B1. Also we included assays for measuring cell cycles and apoptosis by Flow cytometry. In this study, we include these criteria: (1) adequate control, (2) randomization, (3) baseline comparability, (4) blinding, (5) clear description of intervention, (6) outcome measures, (7) adequate statistical analysis, and (8) reproducibility. 
我們假設這個研究的問題的時間很短。我們決定使用長時間的研究,假設某些玻璃內研究失敗的正 面影響取決於他們的長壽和療癒介入的數量。舉例來說,在 Jhaveri 進行的一項研究中,只有經過 長時間才能證明能量療癒對細胞的影響。在我們的實驗中,我們使用了靈氣技術訓練過的 biofield 治療師。靈氣起初是一種來自日本的醫學,現在在世界各地使用。我們使用了 MCF-7 細胞和三名靈 氣訓練治療師,治療師是靈氣導師(2 年經驗)和其他兩名靈氣二級學生(1 年經驗)。由一個沒 有治療經驗的人進行沒有治療過的控制組和安慰劑治療組。我們使用細胞計數的方法,包括使用 Coulter Counter model Z B1 直接的細胞計算。我們也還使用包括通過流式細胞術測量細胞週期 和細胞凋亡的測定。在這個研究中,我們囊括這些條件(1)適當控制,(2)隨機分配,(3)基本對比, (4)盲性,(5) 明確介紹干預措施,(6) 成果,(7) 充分的統計分析,(8)再生性。 

METHODS 方法 Cell line and assays 細胞株及試驗
MCF-7 breast cancer cells were purchased from Clonetics/BioWhitaker (San Diego, CA). Cells were cultured and maintained in 8% CO2 at 37 °C in RPMI 1640 (Invitrogen, Carlsbad, CA), supplemented with 10% heat inactivated FCS (HyClone, Logan, UT), 2 mM l-glutamine, 100 U/ml penicillin, and 100 U/ml streptomycin (Invitrogen, Carlsbad, CA). Standard chemicals were purchased from Sigma (St. Louis, MO). Cell monolayers grown on six-well cell culture plates were used for all of the experiments. The cells were seeded 24 hours before treatment with 10,000 cells per well for all experiments. The cells were analyzed and reseeded each week. Reseeding was done by trypsin, harvesting the cells from plates used in the previous week and planting 10,000 cells per well for the new treatment. This reseeding allowed us to avoid cell overgrowth and still continue the treatment for as many weeks as needed. 人類乳癌細胞(MCF-7)購自 Clonetics / BioWhitaker。細胞培養在 37℃RPMI 培養基中並保持在 8%CO 2 裡,以 10%熱滅活 FES 補充,2 mM l-glutamine, 100 U/ml penicillin, and 100 U/ml streptomycin。標準化學品購自Sigma公司。在六孔細胞培養板上生長的細胞單層用於所有的實驗。 對於所有實驗的 10,000 個細胞在治療前 24 小時接種。每週分析並重新接種細胞,通過胰蛋白酶進 行再次播種,前一周使用的培養板收穫的 10,000 個細胞進行新的處理。這種重新接種使我們避免 了細胞過度生長,並且仍然根據需要繼續治療數週。 

Treatments; participants; protocols 治療;參與者;計畫
 The most important aspect in CAM research is to avoid, during the study of effectiveness, other possible contributing factors ─ e.g., the natural course or cyclic nature of the disease, placebo effect, spontaneous remission, and misdiagnosis ─ all of which are present during the study of CAM with humans and even with animals [12, 13]. The skepticism of the participant could diminish the results as well. Cell models allow us to exclude these factors. As a control, we used untreated cells that had been maintained in the same condition as the treated cells, and placebo healing with an individual who was not trained in the Reiki healing technique. Cells were randomly distributed into control groups and treated groups. The sample size was one well, with three parallel measurements from other wells. The treatments were provided by three Reiki healers: Master and two second level Reiki practitioners. Master Reiki and Reiki practitioners’ second level could be described as post graduate who can teach students and has more knowledge in subject and students with associate degrees who have less knowledge and experience. None of these individuals had previous experience with healing of biological material. The treatments lasted three weeks for 10 minutes each weekday (five days per week). The measurements were taken in the middle of the week. The healers held their hands 10 cm above the cell culture plate, covered by lid and never touched the plate; this distance allowed us to perform the treatment without increasing the temperature of the cells (temperature measurements had been done). Untreated control plates with cells were maintained with the exact same conditions, but without treatment. The same conditions included taken out the control from incubator while the treatment had been conducted and maintained it in the same light\oxygen\CO2\temperature conditions. The results of the treatment were measured with blind experiments - i.e. person who carried out the Flow cytometry and other measuring was not aware which group each sample belongs to but data analysis was not blinded to group. It’s not necessary in blind experiment because it would be done on next level – double blind experiments which we are planning to do in the future. 
輔助替代療法研究中最重要的方面是避免在有效性研究期間,其他可能的影響因素,如疾病的自然 過程或週期性,安慰劑效應,自發緩解和誤診 - 所有這些都是在人類甚至是動物的輔助替代療法 研究。參與者的懷疑也可能會降低結果,細胞模型允許我們排除這些因素,作為對照,我們使用與 處理的細胞保持相同條件的未經處理的細胞,並且與未經靈氣療癒技術訓練的個體進行安慰劑治癒。 細胞隨機分配到對照組和治療組。樣本量很大,有來自其他的三個平行測量,由三名靈氣治療師提供治療,老師和二級靈氣從業者,靈氣導師和二級的靈氣師可以形容為可以教授學生的碩士研究生, 並且在知識和經驗較少的學科和具有副學士學位的學生中具有更多的知識。這些人以前都沒有生物 物質的治療經驗。治療持續三週,每個工作日 10 分鐘(每週 5 天)。在一週中中間進行測量。療 癒者在細胞培養板上方 10 厘米的地方舉起手,蓋上蓋子,從不接觸板子,這個距離使我們能夠在 不增加細胞溫度的情況下進行治療(溫度測量已經完成)。未治療的細胞對照培養板也是如此,保持 完全相同的條件,但沒有治療。相同的條件包括在進行處理的同時從培養箱中取出對照並維持在相 同的光,氧氣,二氧化碳溫度條件。盲性實驗測量治療的結果 - 即進行流式細胞術和其他測量的 人不知道每個樣品屬於哪個組,但是數據分析不是盲性,盲性的實驗是沒有必要的,因為它將在下 一個階段完成 - 我們打算在未來進行雙盲實驗。

 Statistical analysis 統計分析 All experiments were repeated at least three times in the course of this study. Student t-test calculation was used. Standard deviation (±SD) was calculated where appropriate. SD and Student t-test parameters (t, P) are shown in the tables. Additionally, we used ANOVA testing. The Tukey HSD test also had been use to show the difference in repeated measurements.(Additional information on assays, procedure and statistical analyses is available on request.) Cell growth count for cell growth suppression measurements For the cell growth assay, we chose a direct cell count because our experience with cell culture experiments showed that other methods, such as colorimetric assays such as MTT viability assay used in [5], did not have the accuracy of direct counting and most likely could include false positive as well as false negative results. This had been shown in our laboratory by comparison with direct cell count (data not publish). “Six well plates 24 hours prior to treatment were seeded with 0.5 mL of MCF-7 cells in Dulbecco's Modified Eagle Medium (DMEM).”Treatment was carried out as described above. Sample size was 1 well with 3-6 parallel measurements. After the Reiki treatment, the cells were allowed to grow at 37 °C in a CO2 incubator. Cell growth was measured each week, using a Coulter Counter model Z B1 (Coulter Electronic Inc., Hialeah, FL). The baseline was measured in the extra six-well plates before the experiments began. Baseline variability was not significant in any of the experiments (data not shown but available upon request). 
所有實驗在本研究過程中至少重複三次。使用學生 T 檢定計算,標準偏差在適當的時候計算,標準 偏差和學生 T 檢定如表中所示。另外,我們使用變異數分析,多重比較分析檢定也被用來顯示重複 測量的差異。細胞生長計數用於細胞生長抑制測量,對於細胞生長測定,我們選擇了直接細胞計數, 因為我們的細胞培養實驗經驗表明其他方法,如比色測定法,如 MTT 生存力測定法,與直接細胞計 數相對照,這已經在我們的實驗室中顯示,沒有直接計數的準確性,很可能包括假陽性和假陰性結果。在 Dulbecco's Modified Eagle Medium (DMEM)中的 MCF-7 cells,在治療前 24 小時內播種 0.5 mL 在 6 well 的培養盤分隔盤內,如上所述進行處理。樣本大小為 1,並有 3-6 個平行測量。 在靈氣治療後,允許細胞在在 37℃的 CO 2 培養箱中下生長,每週使用 Coulter Counter model Z B1 測量細胞生長,基準測量是在實驗開始之前的 6 well 的培養盤分隔盤內。基線變異在任何實驗中 都不顯著。 

Flow cytometric analysis of DNA content by cell cycle evaluation 藉由流式細胞儀分析 DNA 含量評估細胞週期 Another test used in this research was the flow cytometric analysis of DNA content, measured by the sub G1 and G1 checkpoint of the cell cycle. The most advanced method of cell cycle and apoptosis measurements is flow cytometry. This method has proven to be quick and very accurate. We chose this method for our research. For measurements, cells were harvested; washed twice in phosphate buffered saline (PBS); fixed with 70% ethanol for 30 minutes at room temperature; washed three times with PBS; resuspended in the staining buffer (10 mM PIPES, pH 6.8, 0.1 M NaCl, 0.1% Triton X-100, 2 mM MgCl2); treated with DNase-free RNase I (100 μg/ml) for 30 minutes at room temperature; and stained with propidium iodide (PI) (Molecular Probes, Eugene, OR) at a concentration of 10 μg/ml, according to a prescribed method [14, 15]. Stained cells were analyzed by flow cytometry, using a Beckman Coulter flow cytometer. 
本研究中使用的另一個測試是藉由流式細胞儀分析 DNA 含量評估 sub G1 and G1 checkpoint 細胞 週期,流式細胞術是測量細胞週期和凋亡最先進的方法,這種方法已被證明是快速和非常準確的, 我們選擇這種方法進行研究。用於測量,收穫細胞; 在磷酸鹽緩衝鹽水中洗滌兩次; 在室溫下用 70%乙醇固定 30 分鐘; 在磷酸鹽緩衝鹽水中洗滌三次; 在染色緩衝液中懸浮(10 mM PIPES, pH 6.8, 0.1 M NaCl, 0.1% Triton X-100, 2 mM MgCl2) ; 用不含 DNA 酶的 RNA 酶 I(100μg/ ml)在室 溫下處理 30 分鐘;並按照規定的以 10μg/ ml 的濃度的碘化丙啶方染色。使用 Beckman Coulter 流式細胞儀通過流式細胞術分析染色的細胞。 Annexin V-FITC and PI staining for apoptosis evaluation Annexin V-FITC 和 PI 染色用於凋亡評估 

The growth of any tissue, whether normal or malignant, is determined by the quantitative relationship between the rate of cell proliferation and the rate of cell death. Annexin V (an early stage of apoptosis). Increasing the mechanism that normally regulates cell death, such as apoptosis, has important ramifications for cancer treatment [16]. Therefore, the increasing apoptosis of cancer cells could be considered a healing process because it is the process of returning the cells to the normal regulation process. Cells, both attached (to the plates) and detached, were collected, washed twice in PBS, and doubly labeled with Annexin V-FITC (fluorescein isothiocyanate fused with Annexin V) and PI, according to a prescribed procedure. Cells were seeded in 6 well plates prior treatment. Treatment was carried out as described above. Both adherent and detached cells from untreated and exposed samples were collected, washed twice in PBS and double-labeled with Annexin V and PI according to the manufacturer's protocol (Sigma, St. Louis, MO). Control populations consisted of unstained cells and cells stained with only with Annexin V or PI. Cells were analyzed by flow cytometry using a Beckman Coulter flow cytometer. The fluorescence emission of 10,000 cells /treatment was collected in each channel. [14, 15]. Cells stained only with Annexin V or PI were used as controls. Labeled cells were analyzed by flow cytometry, using a Beckman Coulter flow cytometer. The fluorescent emission of 10,000 cells was collected in each channel. In these studies, the early stages of apoptosis of cancer cells, after Reiki treatment, were evaluated by flow cytometry, using Annexin V-FITC, a fluorescent indicator dye that binds to phosphatidylserine in the cell membrane. 
任何組織的生長,無論是正常還是惡性,都是由細胞增殖速度與細胞死亡率之間的定量關係決定的。 早期凋亡的階段。通常增加調節细胞程序性死亡機制,例如凋亡,對於癌症治療具有重要的影響。 因此,癌細胞凋亡的增加可以被認為是一個癒合過程,因為它是使細胞恢復正常調控過程的過程。 僅用 Annexin V-FITC 或 PI 染色的細胞用作對照。 收集細胞,將其連接並分離,在 PBS 中洗滌兩次,並根據規定的程序用 Annexin V-FITC(與膜聯 蛋白 V 融合的異硫氰酸熒光素)和 PI 雙重標記。在治療前將細胞接種於 6well 的培養盤分隔盤中。 如上所述進行處理。收集來自未處理和暴露樣品的粘附和分離的細胞,在 PBS 中洗滌兩次並根據製 造商的方案用 Annexin V 和 PI 進行雙重標記。對照群由未染色的細胞組成,細胞僅用 Annexin V 或 PI 染色。使用 Beckman Coulter 流式細胞儀通過流式細胞術分析染色的細胞。每個通道收集 10,000 個處理的螢光放射細胞。在這些研究中,在評估靈氣處理後癌細胞凋亡的早期階段使用 Annexin V-FITC(一種結合細胞膜中磷脂酰絲氨酸的螢光指示劑染料)通過流式細胞術。 

RESULTS 結果 
We found that in treated samples, cancer cell growth suppression followed DNA content changes, as measured by cell cycle evaluation and apoptosis using detection of the early stages of apoptosis. Placebo treatment did not statistically differ from the control group (data not shown). The results in Table 1 show the number of cells; results in Tables 2 and 3 and on the flow cytometry charts indicate percentages. In MCF-7 cells, the number of treated cells decreased significantly, 1.5-1.7 fold, with P<0.05 after two weeks of treatment 我們在處理的樣品中發現,癌細胞生長抑制隨 DNA 含量的變化而變化,如通過細胞週期評估和細胞 凋亡使用檢測細胞凋亡的早期階段。表 1 中的結果顯示了細胞的數量;表 2 和表 3 中的結果以及流式細胞術圖表顯示了百分比。在 MCF-7 細胞中,治療細胞數目顯著減少 1.5-1.7 倍,治療兩週後 P <0.05。 Table 1. Suppression of cancer cell proliferation 抑制癌細胞增殖 An evaluation of DNA content revealed that treatment significantly (student’s t-test, P< 0.05 and ANOVA test P<0.002) increased the sub G1 (apoptotic) checkpoint in MCF-7 cells up to 4-23 fold, as shown in Table 2, Fig.1. The Tukey HSD test also show the difference in repeated measurements. DNA 含量的評估中顯示治療顯著(學生 t 檢驗,P <0.05 和方差分析檢驗 P <0.002)使 MCF-7 細胞 中的 sub G1(凋亡)checkpoint 增加 4-23 倍,如表 2 所示, 圖 1。 Tukey HSD 測試也顯示重複 測量的差異。 Table 2. Cell cycle disruption in the MCF-7 cells 在 MCF-7 細胞中的細胞中斷週期 

Figure 1. Cell cycle of MCF-7 by PI flow cytometry measuring. Cells in 6 well plates have been treated by participant 1 during one week, 10 min during every week day, with hands 10 cm above the plates (it preserves the cells from influence of hand temperature). Results were based on the principle that apoptotic cells, among other typical features, are characterized by DNA fragmentation and, consequently, show a loss of nuclear DNA content. Therefore, a fluorochrome, such as PI, which is capable of binding and labeling DNA, makes it possible to obtain results of apoptosis in cancer cells by evaluating the cellular DNA content with flow cytometric analysis and subsequent identification of hypodiploid cells. The percentage of early stage apoptotic cells in a treated culture and evaluated by flow cytometry is shown in Figure 2 and Table 3. The Tukey HSD test also shows the difference in repeated measurements 
圖 1.通過碘化丙啶流式細胞術測量的 MCF-7 的細胞週期。細胞在 6 well 的培養盤分隔盤內,已經 由參與者 1 治療一週,每週 10 分鐘,手在培養盤分隔盤上方 10 公分。結果是基於以下原理:凋亡 細胞以及其他典型特徵,其特徵在於 DNA 片段化,顯示核 DNA 含量的損失。因此,通過能夠結合和 標記DNA的碘化丙啶等螢光色素,通過流式細胞儀分析評價細胞DNA含量,隨後鑑定亞二倍體細胞, 可以獲得癌細胞凋亡的結果。圖 2 和表 3 顯示了經處理的培養物中早期凋亡細胞的百分比並通過流 式細胞術評估.Tukey HSD 測試也顯示了重複測量的區別。 Table 3. Apoptosis of MCF-7 cells after treatment 表 3.治療後 MCF-7 細胞的細胞凋亡 

Figure 2. Apoptosis in MCF-7 cells under Reiki treatment measured by Annexin V-FITC staining with flow cytometry measurement. Cells in 6 well plates have been treated by participant 1 during one week, 10 min during every week day, with hands 10 cm above the plates (it preserves the cells from influence of hand temperature). The cells under treatment expressed phosphatidylserine (PS) at the plasma membrane and binding Annexin V (an early stage of apoptosis). This revealed a significant 1.5 fold (P<0.05) increase in the amount of early stage apoptotic MCF-7 cells, demonstrated by the Annexin V–FITC staining after the treatment. Propidium iodide exclusion was used for MCF-7 cells’ viability determination. Living cells have intact cell membranes and active cell metabolisms that exclude PI viability dye, while cells with damaged membranes or impaired metabolisms allow this dye to enter the cell. 
圖 2.通過流式細胞術測量通過 Annexin V-FITC 染色測量的在靈氣治療下的 MCF-7 細胞中的細胞凋 亡。細胞在 6 well 的培養盤分隔盤內,已經由參與者 1 治療一週,每週 10 分鐘,手放在培養盤分 隔盤上方 10 公分。細胞在質膜上表達磷脂酰絲氨酸(PS)治療下並結合 Annexin V. (凋亡的早期 階段)。這表明在處理後通過 Annexin V–FITC 染色顯示了早期階段凋亡的 MCF-7 細胞量的顯著增 加 1.5 倍(P <0.05)的增加。碘化丙啶用於排除 MCF-7 細胞活力測定。活細胞具有完整的細胞膜 和排除碘化丙啶生存力染料的活性細胞代謝,而具有受損膜或代謝受損的細胞允許該染料進入細胞。 

Figure 3. Apoptosis detection in MCF-7 cells. By Annexin V- FITC, PI staining; A- before treatment, B- after treatment 3 weeks. Cells in 6 well plates have been treated during three weeks, 10 min during every week day, with hands 10 cm above the plates (it preserves the cells from influence of hand temperature). In Figure 3, the X-axis shows the percentage of cells that were labeled by Annexin-FITC (i.e. apoptotic cells) and the Y-axes shows the percentage of cells labeled by PI (i.e. dead cells). In Figure 3A (before treatment), most of the cells were not labeled by either of these dyes; those are shown primarily in sector 3 of the chart. This demonstrated low levels of apoptosis and low numbers of dead cells. In Figure 3B (after 3 weeks treatment), most of the cells were labeled by Annexin-FITC or PI, primarily in sectors 1, 2, and 4. It proves that after prolonged treatment, most of the cancer cells were dead or in an apoptotic state. 
圖 3.由 Annexin V- FITC,PI 染色檢測 MCF-7 細胞中的細胞凋亡。 A-治療前,B-治療後 3 週。細胞在 6 well 的培養盤分隔盤內,已經治療三週 每週 10 分鐘,手放在培養盤分隔盤上方 10 公分。 在圖 3 中,X 軸顯示了被 Annexin-FITC 標記細胞的百分比,Y 軸顯示由 PI 標記細胞的百分比。在 圖 3A 中,大部分細胞不被這些染料中的任何一種所標記; 這些主要在圖表的第三部分顯示。在圖B 中,大部分細胞被 Annexin-FITC 或 PI 標記,主要在 1,2 和 4 中。這證明經過長時間處理後, 大部分癌細胞死亡或處於凋亡狀態。 

DISCUSSION 討論 
We found that exposure of MCF-7 cancer cells to Reiki treatment significantly accelerated and enhanced their apoptosis in vitro. Additionally, energy treatment dramatically inhibited the proliferation of MCF-7 cancer cells. The number of MCF-7 cells in the sub G1 checkpoint increased as well. This cell growth suppression could have resulted from damage to cancer cells from the treatment and from the re-establishment of natural balance in cell proliferation and death (restoration of the apoptotic process). Taken together, the results indicated that energy treatments caused cancer cell growth suppression and death. In addition, energy treatment was effective in activating apoptosis in MCF-7 breast cancer cells. Inhibition of deregulated cell cycle progression in cancer cells is an effective strategy to halt tumor growth. As demonstrated in Table 2 and Figure 1, treatment significantly increased the percentage of sub G1 apoptotic MCF-7 breast cancer cells. The cellular response to stress induced by treatment with anticancer agents is a key determinant of drug or energy activity. There is evidence that cancer cells often have defects in one checkpoint control, making them more vulnerable to inhibition of a second checkpoint and, thereby, enhancing the overall response to treatment [17]. In recent years, a better understanding of the molecular process as the basis of apoptosis has led to novel strategies that could be employed for the treatment of tumors [17]. Experimental data has suggested that it is theoretically possible to drive cells through apoptotic pathways, rather than into cell cycle arrest pathways, with a dramatic increase in the sensitivity of anticancer agents [18]. 
我們發現 MCF-7 癌細胞暴露於靈氣治療中顯著的加速和增強其凋亡。此外,能量治療顯著的抑制了 MCF-7 癌細胞的增殖。MCF-7 細胞裡的 sub G1 checkpoint 數量也增加了。這種細胞生長抑制可能 是由治療對癌細胞的損傷以及細胞增殖和死亡的自然平衡的重建造成的。綜上所述,結果表明能量 處理引起癌細胞生長抑制和死亡。 另外,能量治療對於激活 MCF-7 乳腺癌細胞中的細胞凋亡是有效的。抑制癌細胞中失調的細胞週期 進程是腫瘤生長停止的有效策略。如表 2 和圖 1 所示,治療顯著增加 sub G1 細胞凋亡 MCF-7 乳腺 癌細胞的百分比。抗癌劑治療誘導的關鍵是藥物或能量活性。 證據表明,癌細胞在一個 checkpoint 控制中經常有缺陷,使其更容易受到抑制second checkpoint的影響,從而提高對治療的整體反應。 近年來,對於細胞凋亡基礎的分子過程有了更好理解,已可用於治療腫瘤的新策略。實驗數據表明, 抗癌劑的敏感性顯著增加,可能是通過凋亡途徑驅動細胞,而不是進入細胞週期阻滯途徑。 Possible mechanism of energy healing 能量癒合的可能機制 

For an explanation of the effect of energy healing on isolated cells, we considered a novel theory which combined research findings on biofield medicines and quantum physics, and which included the characterization of endogenous energy fields in health and disease, as well as an understanding of how these energy fields regulate physiologic processes [19]. 
為了解釋能量治療對孤立細胞的影響,我們考慮了一個新穎的理論,該理論結合了生物領域藥物和 量子物理的研究成果,其中包括內源性能量場在健康和疾病中的表徵,以及對如何理解這些能量場 調節生理過程。 

We examined earlier research [20], which was the first to postulate the existence of a unifying organizing principle for all manifestations of life, the existence of a vectorial "biological field" or "biofield." According to Gurwitch, this field performed the organizing function for all vital processes taking place within its realm. We also considered research that showed the body’s ability to generate both classical force fields and non-classical potential fields [19]. In an attempt to understand and validate CAM biofield therapies, such as Reiki, Qigong and Therapeutic Touch, investigators measured the low electromagnetic frequencies (LEMF) emitted by the body in near-field conditions (near the body) using both physical and biologic detectors as had been shown in the research of Seto [21] and Zimmerman [22]. The mechanism of LEMF affecting biological tissues has been studied in the former Soviet Union and, recently, in the United States and Europe. It has been shown that the cells begin to emit LEMF waves after the transition from a steady state to an altered homeostasis state. This endogenic LEMF, generated by signals from regulatory endogenic LEMF, helps to return cells to their normal homeostasis state [23-25]. If, for some reason, cells are not able to produce the endogenic LEMF, it is possible to help them return to the normal homeostatic state by resonance with exogenic LEMF exposure from electromagnetic devices or LEMF from persons [26-29]. In the future we’re supposed to repeat this study and additionally measure the LEMF. 
我們研究了早期的研究,這是第一個假設存在所有生命表現的統一組織原則,“生物場”或“生物 場”的存在。根據 Gurwitch 的說法,這個領域為所有在其領域內發生的重要進程執行組織功能。 我們也考慮顯示人體能夠產生古典力場和非經典勢場的研究。為了理解和驗證輔助替代療法生物場 的治療方法,如靈氣,氣功和治療性觸覺,研究人員使用物理和生物檢測器測量身體在近場條件下 (身體附近)發出的低電磁頻率(LEMF)已經在 Seto 和 Zimmerman 的研究中得到證實。低電磁頻 率影響生物組織的機制已經在前蘇聯和在美國和歐洲進行了研究。已經顯示,細胞從穩定狀態轉變 為改變體內平衡狀態之後開始發出低電磁頻率波。這種由內源性低電磁頻率信號產生的內源性低電 磁頻率有助於使細胞恢復正常的體內平衡狀態。如果由於某些原因,細胞不能產生內源性低電磁頻 率,則可能通過與來自人的電磁裝置或低電磁頻率的外源性低電磁頻率暴露共振來幫助它們恢復到 正常的內穩定狀態。將來我們應該重複這個研究以及另外測量低電磁頻率。 

Thus, the healing effects of Qigong, Reiki, Therapeutic Touch, etc., could result from the action of exogenic LEMF (classical, which caused effects during contact treatment, and non-classical, which caused effects during distant treatment) emitted from a healer and resonated within the endogenic cell’s LEMF. Also, we could not exclude the possibility that exogenic LEMF could damage cancer cells by itself (like chemotherapy) and not exclusively through a recovery mechanism. This is possible due to the differences in frequencies between normal cells and cancer cells [30-32]. This difference has been noted recently in a method called “photo acoustic detection,” a technique utilized to identify the characteristic sound of melanoma cells, which differs from the sound of normal cells [33]. 
因此,氣功,靈氣,治療性觸覺等的治療效果可能是由外源性低電磁頻率(在接觸治療中產生效果 的經典,以及在遠端治療中產生效果的非經典)並在內生細胞的低電磁頻率內發生共振。此外,我 們不能排除外源性低電磁頻率可能會自身損傷癌細胞(如化療)的可能性,以及不能完全通過恢復 機制。正常細胞和癌細胞之間的頻率差異,他是有可能的。這種差異最近已經被稱為“光聲檢測” 的方法,該方法被用來識別黑素瘤細胞的特徵性聲音,其與正常細胞的聲音不同。 

The present research, like the research of Jhaveri [7] shows that a significant difference between treated and control groups appeared only a week or two after the energy treatment began. However, if we consider the healing effect of the LEMF emitted by the healer, and if the healer emits a strong electromagnetic field, the results of treatment could be rapid. Additional research [21] has shown that 8% of tested subjects exhibited a particularly strong bio-magnetic field, at least 1,000 times greater than that of the normal human bio-magnetism. The prompt positive results of energy healing, demonstrated in research, could be attributed to the extraordinary electromagnetic field emitting from the participating healers. For healers who do not emit such strong energy, much more time is needed to show measurable effects. This may explain why certain studies of bioenergetics and Qigong [2, 4, 5] failed to show any difference between treated and control groups. These studies included a limited number of healing sessions and a short duration between the healing activity and the measurement of results. 目前的研究,如 Jhaveri 的研究表明,治療組與對照組在治療開始後一兩週之間才出現顯著差異。 然而,如果我們考慮療癒者施放的低電磁頻率治療效果,而且如果治療者發出強大的電磁場,則治 療結果可以是快速的。另外的研究說明,8%的受試者表現出特別強的生物磁場,至少比正常的人 類生物多出 1000 倍磁性。研究表明,能量癒癒的迅速積極結果可歸因於參與治療者發出的非凡的 電磁場。對於不常輸出如此強大能量的治療者來說,需要更多的時間來顯示可測量的效果。這也許 可以解釋為什麼某些生物能學和氣功研究沒有顯示治療組和對照組之間的差異。這些研究包括有限 的療癒期間和短時間的療癒活動與結果測量。 

Strengths and weaknesses in relation to other studies 與其他研究的優點和缺點 
A systematic review of published healing research [34] indicates that many studies fail to fulfill important quality criteria, including: (1) adequate control, (2) randomization, (3) baseline comparability, (4) blinding, (5) clear description of intervention, (6) outcome measures, (7) adequate statistical analysis, and (8) reproducibility. In the present study, all of the above criteria were met [34]. In the study, every outcome parameter was quantifiable; thus, the measurements did not rely on the subjective assessments of pain relief or of a general state of health that were common in other studies. The control, which was handled under the same conditions, excluded the possibility that the decrease of cancer cells’ growth and damage were the result of any other reasons except, perhaps, the practitioners’ influences (such as environmental conditions). Although this study was blind (the individual who performed the measurements did not know whether the samples belonged to the Reiki group or the control group), double-blind experiments will be needed to prevent healing practitioners from knowing to which group the sample belonged. Other limitations were: the data analysis may not have been blinded to the study groups. 
已發表的癒癒研究評價表明許多研究未能達到重要的質量標準,包括:(1)適當的控制,(2)隨機 化,(3)基線可比性,(4)盲性,(5)明確的描述(6)結果測量,(7)充分的統計分析,(8)重 現性。在本研究中,所有上述標準都得到滿足。 在研究中,每個結果參數都是可以量化的; 因此,測量不依賴於其他研究中常見的疼痛緩解或一般 健康狀況的主觀評估。在相同條件下掌控的對照組排除了癌細胞生長和損傷的減少,可能是由於排 除了可能的影響(例如環境條件),雖然這項研究是盲性的(進行測量的個人不知道樣本是屬於靈 氣組還是對照組),但是需要雙盲實驗來防止療癒者知道樣本屬於哪個組。其他限制是:數據分析 的研究小組可能不是盲性。 

CONCLUSION 結論 
The Reiki treatment from trained healers may damage cancer cells or may restore the organism to a state in which natural processes can occur. Like chemotherapy drugs, these biofields induce cells to undergo apoptosis and result in cell cycle damage. However, the amount of time needed for biofields to effect changes may be longer than the time required for chemotherapy. Therefore, more rigorous, double-blind, placebo-controlled and perhaps animal-based experiments are needed to prove our hypothesis and to show the effectiveness of Reiki enhanced biofields on reducing cancer cell growth throughout the body. However, the results of this pilot study may help the scientific community to understand the effects of Reiki biofield energy on cancer cells. This procedure may still be useful in decreasing the growth of cancer cells in cellular research, in animal research and perhaps in humans.
來自訓練有素的治療者的靈氣療法可能會損傷癌細胞,或者可能使有機體恢復到可能發生自然過程 的狀態。與化療藥物一樣,這些生物領域誘導細胞發生細胞凋亡並導致細胞週期損傷。然而,生物 場所需要的時間變化可能比化療所需的時間更長。因此,需要更嚴格的雙盲,安慰劑對照,也許動 物為基礎的實驗來證明我們的假設,並顯示靈氣增強生物場可減少整個身體癌細胞生長的有效性。 

Table 1. Suppression of cancer cell proliferation 抑制癌細胞增殖
Groups Cells counts (mean ± standard deviation) 1 week 2 weeks 3 weeks MCF-7 control 42900 ±1680 36100 ± 4690 42770 ± 1573 MCF-7 treated 40300 ± 1720 24100 ±4320 30800± 2190 % to control 93 66.7 72.0 t; P (Student’s test) 1.92; < 0.13 3.26; <0.03 7.61; < 0.002 F;P (ANOVA) 18.77; 0.0001 HSD=3.62 HSD.01= 4.71 1w&2w ( in thousands) 16.2 1w&3w ( in thousands) 9.5 2w&3w 6.7 MCF-7 control (repeat) 37300 ± 2070 42400 ± 6070 35100 ± 2210 MCF-7 treated (repeat) 35200 ± 1450 32000 ± 2290 23100 ± 2530 % to control 94.4 75.5 65.8 t; P (Student’s test) 1.24; <0.28 2.49; <0.06 6.22; <0.003 F;P (ANOVA) 12.45:0.0002 HSD=3.76 HSD.01= 5.05 1w&2w ( thousands) 3.2 1w&3w(thousands) 12.1 2w&3w (thousands) 8.9 Table 2. Cell cycle disruption in the MCF-7 cells MCF-7 細胞中的細胞週期中斷 Cells in Sub G1 checkpoint (%) Parameter 1 week 2 weeks 3 weeks Sub G1 control 1.2 ± 0.2 1.2 ± 0.6 5.1 ± 0.5 Sub G1 treated 28.4 ± 16.4 13.5 ± 1.2 19.8 ± 3.3 % to control 2366.7 1125 388 t; P (Student’s test) -2.87; <0.05 -16.1; <0.001 -7.59; <0.002 F;P (ANOVA test) 7.8;0.0018 HSD=4.17 HSD.01= 5.91 1 w&2 w 14.9 1w&3w 8.6 2w &3 w 6.3 Table 3. Apoptosis of MCF-7 cells after treatment. MCF-7 cells 治療後的細胞凋亡 Cells Apoptotic cells (cells with Annexin –FITC), after 1 week of treatment, (%) MCF-7 control 1.37 ± 1.48 MCF-7 treated 35.8 ±4.94 t; P( Student’s test)11,3; <0.001 Figure 1. Cell cycle of MCF-7 by PI flow cytometry measuring. Cells in 6 well plates have been treated by participant 1 during one week, 5-10 min during every week day, with hands 2-4 inch above the plates (it preserve the cells from influence of hands temperature). 
圖 1.通過碘化丙啶流式細胞術測量的 MCF-7 的細胞週期。細胞在 6 well 的培養盤分隔盤內,已經 治療一週每週 5-10 分鐘,手放在培養盤分隔盤上方 2-4 英寸(其保持細胞免受手部溫度的影響)。 

Figure 2. Apoptosis in MCF-7 cells under Reiki treatment measured by Annexin V-FITC staining with flow cytometry measurement. Cells in 6 well plates have been treated by participant 1 during one week, 5-10 min during every week day, with hands 5-10 cm above the plates (it preserve the cells from influence of hands temperature). 
圖 2. 用流式細胞術測量 Annexin V-FITC 染色測量的 MCF-7 cells 在靈氣治療下的細胞凋亡。細 胞在 6 well 的培養盤分隔盤內,已經治療一週,每週 5-10 分鐘,手放在培養盤分隔盤上方 5-10cm (其保持細胞免受手部溫度的影響)。 

Figure 3. Apoptosis detection in MCF-7 cells. By Annexin V- FITC, PI staining; A- before treatment, B- after treatment 3 weeks. Cells in 6 well plates have been treated during three weeks, 5-10 min during every week day, with hands 5-10 cm above the plates (it preserve the cells from influence of hands temperature) 
圖 3.MCF-7 細胞中的細胞凋亡檢測。Annexin V-FITC, PI 染色; A-治療前,B-治療後 3 週。細胞在 6 well 的培養盤分隔盤內,已經治療一週,每週 5-10 分鐘,手 放在培養盤分隔盤上方 5-10cm(其保持細胞免受手部溫度的影響)。 

https://www.linkedin.com/pulse/20140312224018-138727290-reiki-can-treat-cancer-cells

張育菁
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