Effect of Subtle Energy Transmission and Tao Calligraphy Mindfulness Practice on Telomere Length in Peripheral Blood Leukocytes A Follow-up Pilot Study
Phase: NA
Conditions: Telomere Length, Quality of Life
Sponsor: Sha Research Foundation

1 
 
 
STUDY TITLE: 
Effect of Subtle Energy Transmission and Tao Calligraphy 
Mindfulness Practice on Telomere Length in Peripheral Blood 
Leucocytes 
 
 
A Follow-up Pilot Study 
 
 
PRINCIPAL CLINICAL INVESTIGATOR: 
Name: XX, MD, FRCS 
Department: Sha Research Foundation, San Francisco, California 
 
 
PRINCIPAL RESEARCH INVESTIGATOR: 
Name: XX, PhD 
Department: Sha Research Foundation, San Francisco, California 
 
 
VERSION DATE: 
Version 3 of 10th April 2023 
 
 
RELATED STUDIES:  
None 
 
 
Note:  Names of research participants were removed as per instructions on Clinical Trials Gov 
Website. 

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1.0 
Purpose and rationale of the study: 
 
OBJECTIVES 
The study is intended to measure the effects and determine the efficacy of Tao 
Calligraphy mindfulness practice together with subtle energy transmission on increasing 
length of telomeres in peripheral blood leucocytes. In our previous studies, the 
participants who regularly practiced reported a decrease in symptoms of their illness as 
perceived subjectively, an improvement in laboratory results or other tests and observed 
signs of progress reported by treating clinician and expressed an increase in well-being 
as measured by standardized scientific questionnaires.  
 
 
HYPOTHESIS 
 
The research null hypothesis is that individuals who receive subtle energy transmission 
for improving telomere length in peripheral blood leucocytes, and who will regularly 
practice mindfulness with Tao Calligraphy for 3 months, will have no significant change 
in telomere length in follow-up analyses and will show no improvement of well-being or 
clinical status as measured by standardized scientific questionnaire Rand SF 36(s) at 3 
and 9 months. 
 
Tools such as the Minitab version 14 and PSPP/SPSS will be used to analyze the data 
from laboratory assessment and Research Questionnaires at the start and at 3 months 
and at the end of 9 months of participation in the study. Statistical test such as Anova 
will be used to evaluate the null hypothesis. The p-value will represent how unlikely the 
observed data would be if the null hypothesis were actually true and we will use them 
to reach conclusions.  We set the confidence level at 95% and if we receive p < 0.05, 
then H0 is rejected.   
 
The correlation coefficient will be used to determine any correlation between various 
factors (e.g., effects of age, sex, length, and frequency of mindfulness practices and 
others on outcome) and regression analysis to determine the relation of independent 
and dependent variables.  
 
 
BACKGROUND 
 
Markers of Longevity 
Mammalian development involves an exquisite choreography of cell division, 
differentiation, programmed cell death, and senescence that directs the transformation 
of a fertilized egg to a mature organism containing on the order of 40 trillion cells in 
humans.  When the molecular synthesis, repair or replacement systems are unable to 
maintain the positive balance that existed prior to reproductive maturity, a tipping 

3 
 
point is reached, leading to the universal aging process associated with susceptibility to 
disease and ultimately death. [1] [2] [3] [4] [5] 
There is a dynamic interplay between genetic and environmental variations in the 
development of individual differences in health [6] and, hence, longevity.  The genetic 
component influence on lifespan in humans is estimated to be about 25%, e.g., by 
comparing the age of death of monozygotic and dizygotic twins, and this genetic 
component tends to be higher at older ages and in males. [7] [8] [9]  
Telomere length as a biomarker of aging [10] 
Telomeres are repetitive (TTAGGG)n sequences located at the ends of linear 
chromosomes.  Telomere length shortens with age.  Shorter telomeres have been 
associated with increased incidence of diseases and poor survival. The rate of telomere 
shortening has been found to increase or decrease in association with specific lifestyle 
factors.  
According to the American Federation for Aging Research [11], a true biomarker of 
aging should meet the following criteria:  
1) predict remaining life expectancy better than chronological age,  
2) monitor mechanisms underlying the aging process but not a specific disease,  
3) be subject to repeated tests without harming the individual,  
4) be testable in both laboratory animals and humans.  
 
While telomere length (TL) satisfactorily meets criteria 3 and 4, compliance with criteria 
1 and 2 is less conclusive as there are significant inconsistencies observed among 
studies [12] .    
The phenomenon of age-related telomere shortening is complex and biological 
mechanisms underlying this process are not yet definitively established. In particular, it 
is still unclear whether telomeric aging reflects a mitotic clock-like process or 
alternatively, a biomarker of stress or a biological mechanism that transfers stress-
associated signals to the cell [13] [14].  
Despite the uncertainty of the usefulness for estimating the human biological age, TL 
currently remains one of the most widely used biomarkers in epidemiological and 
clinical studies of aging. In recent years, TL has also increasingly been investigated as a 
potential biomarker in personalized medicine [15], especially in combination with other 
measures, as described by Ahadi et al., 2020 [16].   
Other clinical markers 
In a longitudinal study of 106 pre-diabetic individuals aged 29 to 75 years, aging 
markers whose levels changed over a short timeframe of 2–3 years were identified that 
may ultimately be useful in monitoring and intervening in the aging process [16].  To 
understand how molecules that correlated with age in a population compared to their 
trends in individuals, clinical laboratory markers were examined. For each quarterly 
visit, proteomics and metabolomics on plasma samples and transcriptome analysis on 
material from peripheral blood mononuclear cells and targeted cytokine assays using 
serum were performed. Nasal and gut microbiomes were analyzed using 16S rRNA 
sequencing, providing information at the genus level, and exome sequencing was 
performed once, using DNA from peripheral blood mononuclear cells. In addition, 51 

4 
 
clinical laboratory tests were acquired on each visit.   Although overall phenotypic age 
increased with age as expected (slope = 1.07), the individual phenotypic age slopes 
differed among the different participants, with 15 participants even showing negative 
values (Fig. 3e). To determine whether lifestyle could affect age-associated clinical 
markers directly, lifestyle, BMI and medication data for participants who deviated from 
the population trend were examined. The results suggest that individuals were aging at 
different rates as well as potentially through different biological mechanisms and 
lifestyle interventions. 
Suggested minimally invasive sample collection methods for DNA (the 1) other in 
pilot phase) 
1) Venous blood buffy coat (“gold standard”) 
2) Salivai 
 
Suggested DNA measures (the 1) in pilot phase)  
1) Telomere length 
2) DNA Methylation ii 
Suggested laboratory measures for phenotypic age calculationiii  (not in pilot phase) 
3) From venous blood: albumin, creatinine, glucose, log (C-reactive protein), lym-
phocyte percent, mean cell volume, RDW, ALKP and white blood cell count 
 In an evaluation of minimally invasive sample collection methods for telomere length 
measurement, Oragene saliva was moderately correlated (ρ = 0.48, p = 0.002) and the 
most similar in size to buffy coat from venous blood, considered the “gold standard.” 
[17] 
2 Cross-sectional studies have revealed differences in telomeres and DNA methylation 
associated with age [22]. The latter has led to a description of a ‘molecular clock’ that is 
associated with chronological and biological age [19] [22]. 
 
3 Phenotypic age calculated using the equation provided by Liu et al. [22] [22] that uses 
chronological age and nine biomarkers, including albumin, creatinine, glucose, log (C-
reactive protein), lymphocyte percent, mean cell volume, RDW, ALKP, and white blood 
cell count. 
 
 
Lifestyle Intervention Meditation):  proposed biological mechanisms and study design 
implications 
From 2011 Intensive meditation training, immune cell telomerase activity, and 
psychological mediators Psychoneuroendocrinology Volume 36, Issue 5, Pages 664-681 
Telomerase is the cellular enzyme primarily responsible for telomere length and 
maintenance. In the first study to link meditation and positive psychological change 
with telomerase activity the data suggest that increases in perceived control and 
decreases in negative affectivity contributed to an increase in telomerase activity, with 
implications for telomere length and immune cell longevity.  

5 
 
Methods 
Retreat participants (n = 30) meditated for ∼6 h daily for 3 months and were compared 
with a wait-list control group (n = 30) matched for age, sex, body mass index, and prior 
meditation experience. Retreat participants received instruction in concentrative 
meditation techniques and complementary practices used to cultivate benevolent 
states of mind (Wallace, 2006). Psychological measures were assessed pre- and post-
retreat. Peripheral blood mononuclear cell samples were collected post-retreat 
(baseline/pre-retreat was not collected) for telomerase activity. Because there were 
clear, a priori hypotheses, 1-tailed significance criteria were used throughout. 
Results 
Telomerase activity was significantly greater in retreat participants than in controls at 
the end of the retreat (p < 0.05). Increases in Perceived Control, decreases in 
Neuroticism, and increases in both Mindfulness and Purpose in Life were greater in the 
retreat group (p < 0.01). Mediation analyses indicated that the effect of the retreat on 
telomerase was mediated by increased Perceived Control and decreased Neuroticism. 
In turn, changes in Perceived Control and Neuroticism were both partially mediated by 
increased Mindfulness and Purpose in Life. Additionally, increases in Purpose in Life 
directly mediated the telomerase group difference, whereas increases in Mindfulness 
did not. 
[25] C. Dal Lin et al.: 2021, Rapid changes of miRNAs-20, -30, −410, −515, −134, 
and −183 and telomerase with psychological activity: A one year study on the 
relaxation response and epistemological considerations.  Journal of Traditional and 
Complementary Medicine 
Telomerase is a cellular enzyme that adds the necessary telomeric DNA (T2AG3 repeats) 
to the 3 ′-end of the telomeres, protecting their degeneration.74 The activity of this 
enzyme represents a marker of cellular aging75 and is implicated in aging-associated 
diseases.76 However, compared to telomere length, telomerase function seems to 
correlate more faithfully with stress-related psychological mechanisms.75 Stress leads 
to a decrease in its activity77 while relaxation favors its functioning,78 as our results 
appear to confirm. 
The telomerase activity (Fig. 3) appeared to increase during the RR sessions (p < 0.01 
Wilcoxon test at every time point) both in patients and in healthy volunteers. No 
significant variation was found in the CONTROLS (p > 0.05 Wilcoxon test at every time 
point). The basal values (before and after 20 min) of telomerase activity were 
comparable in all groups and were merged into a single starting basal point. Micro-
RNAs of cardiovascular interest, involved in cell senescence and in the NF-κB network 
(miRNAs −20, −30, −410, −515, −134, and −183), and the activity of telomerase in 
peripheral blood mononuclear cells (PBMCs) were measured in the serum of patients 
with ischemic heart disease (and healthy controls) immediately before and after a 
relaxation response session (RR), three times (after the baseline), in one year of follow-
up. Results: miRNA-20 and -30 levels and PBMCs-telomerase activity increased during 

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the RR while the −410 and −515 levels decreased. During the RR sessions, both miRNA-
134 and -183 decreased.  
The orientation of mental processes, either toward stress or toward relaxation, can impact 
cellular ageing through at least three main recognized pathways: the immune system,70 the 
oxidative balance,71 and the activity of telomerase.72  Although telomere  length is implicated in 
cellular ageing, the evidence suggesting telomere length is a biomarker of ageing in humans is 
equivocal,73 if not invalid.57 
 
As stated, we previously assessed malondialdehyde (oxidative-stress marker) and galectin-3 
(immune system-inflammatory marker) levels as aging58,59 and cellular senescence 
markers,60, 61, 62, 63 demonstrating a decrease in the course of RR.64 Numerous studies in the 
literature and our previous studies12,19 have shown that RR causes a decrease in the levels of 
inflammatory cytokines, stress hormones, inflammatory genes 
expression,19 and epigenetic markers of ageing.20 
 
From 2017 Effect of heartfulness meditation on burnout, emotional wellness, and telomere 
length in health care professionals, Journal of Community Hospital Internal Medicine 
Perspectives, [26] 
 
ABSTRACT Background: Burnout poses significant challenges during training years in residency 
and later in the career. Meditation is a tool to treat stress-related conditions and promote 
wellness. Telomere length may be affected by burnout and stress. However, the benefits of 
meditation have not been fully demonstrated in health care professionals. Objective: We 
assessed the effects of a 12-week ‘Heartfulness Meditation’ program on burnout, emotional 
wellness, and telomere length in residents, faculty members, and nurses at a large community 
teaching hospital during the 2015–16 academic year. Methods: All subjects completed a 
baseline Maslach Burnout Inventory (MBI) and Emotional Wellness Assessment (EWA) at the 
beginning of the study. Meditators received instructions in Heartfulness Meditation. At week 
12, subjects completed a follow up MBI and EWA scores. Salivary telomere length was 
measured at baseline and week 12. Results: Twenty-seven out of a total 155 residents (17.4%) 
along with eight faculty physicians and 12 nurses participated in the study. Thirty-five enrolled 
as meditators and 12 as controls. At 12 weeks, the meditators had statistically significant 
improvement in all measures of burnout and in nearly all attributes of EWA. Controls showed 
no statistically significant changes in either burnout or emotional wellness scores. Relative 
telomere length increased with statistical significance in a younger subset of meditators. 
Conclusion: Our results indicate that meditation offers an accessible and efficient method by 
which physician and nurse burnout can be ameliorated and wellness can be enhanced. The 
increased telomere length is an interesting finding but needs to be confirmed with further 
research. Abbreviations: EWA: Emotional wellness assessment; MBI: Maslach burnout 
inventory; EE: Emotional exhaustion; DP: Depersonalization. 
 
From: Exploring Epigenetic Age in Response to Intensive Relaxing Training: A Pilot Study to 
Slow Down Biological Age    

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Abstract  DNA methylation (DNAm) is an emerging estimator of biological aging, i.e., the often-
defined “epigenetic clock”, with a unique accuracy for chronological age estimation 
(DNAmAge). In this pilot longitudinal study, we examine the hypothesis that intensive relaxing 
training of 60 days in (20) patients after myocardial infarction and in (10) healthy subjects may 
influence leucocyte DNAmAge by turning back the epigenetic clock. Moreover, we compare 
DNAmAge with another mechanism of biological age, leucocyte telomere length (LTL), and 
telomerase. DNAmAge is reduced after training in healthy subjects (p = 0.053), but not in 
patients. LTL is preserved after intervention in healthy subjects, while it continues to decrease 
in patients (p = 0.051). The conventional negative correlation between LTL and chronological 
age becomes positive after training in both patients (p < 0.01) and healthy subjects (p < 0.05). 
In our subjects, DNAmAge is not associated with LTL. Our findings would suggest that intensive 
relaxing practices influence different aging molecular mechanisms, i.e., DNAmAge and LTL, with 
a rejuvenating effect. Our study reveals that DNAmAge may represent an accurate tool to 
measure the effectiveness of lifestyle-based interventions in the prevention of age-related 
diseases. 
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psychological activity: A one year study on the relaxation response and epistemological 
considerations," Journal of Traditional and Complementary Medicine, vol. 11, no. 5, p. 409–
418, 2021.  
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2011.  
 
Chinese calligraphy 
 
Chinese calligraphy handwriting is a dynamic process of integrating visual spatial 
awareness, cognitive planning and motor skills that is maneuvered with a brush to follow 
defined configurations of characters. [1]. There have been growing empirical studies of 
Chinese calligraphic handwriting that resulted in improvements in visual attention and 
span, increased mental concentration [2], confirming it as an effective treatment for 
psychosomatic conditions and post-traumatic hyperarousal symptoms [3] and exhibited 
significant effects on hypertension and type 2 diabetes [4] [5]. Leisure activities occurring 
later in life that includes calligraphy may inhibit cognitive decline [6]. 
 
Tao Calligraphy is a unique branch of Chinese calligraphy (Yi Bi Zi) in that it is 
characterized by one-stroke writing. Every character is written with one continuous 
stroke with the brush always in contact with the paper. [7] Results from studies 
confirmed efficacy in tracing in post-acute rehabilitation setting in that patients reported 
less incontinence, shortened duration of hospital stay, and an increase in overall well-

10 
 
being. [8] Retrospective analysis of data exhibited improvement in general wellbeing, an 
increase in optimism and energy level, as well as improvement of their symptoms. [9] 
According to a study measuring the effects of calligraphy tracing meditation with mantra 
chanting (repeated sound or word to aid with concentration spoken out loud or silently) 
of spiritual practitioners, the results exhibited statistically positive improvement in 
Physical Functioning, Role Limitations due to physical health problems; Role Limitations 
due to Personal or Emotional Problems; Energy / Fatigue; Emotional Well-being; Social 
Functioning; Bodily Pain; General Health. [10] In the prospective follow up study of these 
subjects, following 6 months they continued to improve in functioning in the above-
mentioned areas. The authors concluded that tracing calligraphy and mantra chanting 
was simple to follow, well tolerated, and no complications arose during the study. [11 
 
 
 
 
2.0 
RATIONALE  
The study aims to acquire both basic and applied knowledge to which the primary 
purpose is gain the measurable data of individuals to mindfulness practice with Tao 
Calligraphy together with subtle energy transmission, specifically focused on the 
lengthening of telomeres in peripheral blood Leukocytes. Thus, proposing another 
method of prolonging life expectancy that could be integrated into conventional medical 
treatment and care.  
 
 
THEORETICAL FRAMEWORK 
Tao Calligraphy art has been intensively studied at Sha Research Foundation in the US 
and Canada since 2013, producing significant results in improving overall well-being and 
an efficacious intervention for individuals with chronic conditions. [1]. [2]. [3]. [4]. This is 
a unique style of moving meditation, where mindfulness (heightened awareness) is 
achieved by the combination of movement and focus on Calligraphy art. In this practice, 
the subjects trace the lines of calligraphy with fingers and simultaneously say loudly or 
silently an affirmation (mantra), which enables them to achieve deep concentration 
during wakefulness. 
No risks or negative side effects of the transmission of pure energy given to human 
participants or meditation with Tao Calligraphy have been observed in these pilot 
projects and formal studies. 
 
[1]. P. Hudoba, C. Hamilton, E. A. Yamashiro, and Jaime Chow, Meditation using 
Calligraphy and Chanting Mantra Improves Quality of Life – a Two years 
study, European Journal of Integrative Medicine, Volume 48, 2021, 101965, ISSN 1876-
3820, https://doi.org/10.1016/j.eujim.2021.101965. 

11 
 
[2]. K. Balonwu, Comparative psychometric studies and questionnaires on the effects of 
Tao Calligraphy Tracing Meditation in patients with Unipolar Depression after 25 hours 
and 50 hours., European Journal of Integrative Medicine, Volume 48, 2021, 101964, ISSN 
1876-3820, https://doi.org/10.1016/j.eujim.2021.101964. 
[3]. Tao Calligraphy and Mantra Chanting Improves EORTC QLQ-30 Quality of Life Scores 
for Women Diagnosed with Breast Cancer, Society for Integrative Oncology (SIO) 2021 
Annual Conference, Sept. 24-26, Baltimore, MD, USA.  
[4]. Tao Calligraphy Tracing Meditation with Mantra Chanting Improves Scores of McGill 
Pain Questionnaire (SF-MPQ) in 3 months, Society for Integrative Oncology (SIO) 2021 
Annual Conference, Sept. 24-26, Baltimore, MD, USA.  
 
The proposed research is a pilot project, based on experiential data obtained from 
several previous pilot and formal studies by our research teams and researchers from 
other institutions. It focuses on the effects of transmission of pure energy (blessing) 
and meditation on improving the length of human telomere in peripheral blood 
leucocytes. We believe, the study will provide further data to offer an integrative 
intervention for individuals seeking additional treatment that is of minimal risk and to 
maximize benefits to address condition. 
 
 
METHOD 
 
This study will determine the effects of transmission of a subtle energy and mindfulness 
practice with Tao Calligraphy as measured through subjective and objective evaluations 
of changes in the subjects by Laboratory assessment of Peripheral blood Leucocytes 
Telomere length and subjective changes in RAND SF 36 Questionnaires. 
 
 
Design of the Study 
 
The study will be performed as a Pilot follow-up study, should the 0 Hypothesis be 
successfully rejected, this study will be followed by a formal Double Blind Randomized 
Crossover Study.  
We plan subjects may enter the study as a group and will have measurements done 
within short time frame from each other at beginning and at the end of 3 and 9 months. 
 
As this is a pilot study, there will be no randomization into treatment and control group 
and all subjects will receive a transmission and then will be practicing Tao Calligraphy 
mindfulness daily during the 9 months.  
 

12 
 
Every subject who is accepted into the study, will have blood sample taken for analysis 
and will fill Rand SF 36 Questionnaire. Afterwards, will receive transmission of subtle 
energy by Zhi Gang Sha (or his designee) and will be instructed on how to practice 
mindfulness with Tao Calligraphy. 
 
Every subject who is accepted into the study, will receive prior to the inception of 
practice transmission of pure energy (blessing) and will be instructed on how to practice 
meditations with Tao Calligraphy. 
 
The study is conducted on an outpatient basis and includes: 
 
- 
a telephone discussion (clarification of inclusion/exclusion criteria)  
- 
signing of the consent/information; instruction in the study design and 
practicing the Tao Calligraphy-tracing meditation; first data collection 
questionnaires (Time 0) 
- 
regular weekly meditation practices on Zoom with participants in the treatment 
arm 
- 
Laboratory assessment of telomere Length in peripheral blood Leucocytes at 
the entry time point (Time 0), at the end of 3 months (Time 1), and at the end of 
9 months (Time 2) 
- 
Completion of the questionnaires by the participants at the entry time point 
(Time 0), at the end of 3 months (Time 1), and at the end of 9 months (Time 2) 
- 
Completion of assessments by a Study principal and co-investigators  
 
 
Data collection:  
 
A Laboratory Assessment: 
Laboratory assessment of Telomere Length in Peripheral Blood Leucocytes  
 
Blood samples will be drawn at DAP accredited institution in British Columbia (Life Labs 
or Vancouver Coastal Health) or another accredited institution in Canada and will be 
promptly sent by currier to Repeat Diagnostics, Suite 309 – 267 West Esplanade North 
Vancouver, BC V7M 1A5 Canada for Telomere Assessment. 
Reports will be issued to clinical Principal Investigator Dr. Peter Hudoba  
 
All participants will have blood samples drawn at the entry time point (Time 0), at the 
end of 3 months (Time 1), and at the end of 9 months (Time 2). 
 
 
 
 
 
 

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B Improvement of general well-being or clinical well-being as measured by 
standardized questionnaires  
 
These questionnaires will be used in the study: 
1. John Ware’s SF-36 Quality of Life questionnaire 
2. Simple Follow up Questionnaire of our design 
 
All participants will complete questionnaires at the entry time point (Time 0), at the 
end of 3 months (Time 1), and at the end of 9 months (Time 2). 
 
 
Study Execution 
 
The study will be performed by collecting enrollment data and medical data directly from 
patients personally, over the phone, via email, or on-line (Zoom). The research 
Questionnaires will be filled on-line using personal ID Codes – to ensure confidentiality. 
The codes will be emailed by the PI to each of the participants individually. 
 
Data collection, data handling, and data assessment will be the responsibility of the Sha 
Research Foundation’s research team members who have a medical, nursing, and/or 
scientific background, and will be performed through personal delivery, by email and 
online collection. 
Members of the research team will NOT administer any transmission of pure energy 
(blessing) or any form of medical support to participants of the study. 
 
Transmission of pure energy and instruction of subjects on how to practice meditations 
with Tao Calligraphy will be the sole responsibility of Universal Soul Service Corp. 
The transmission of pure energy (blessing) will be done to participants for free by Tao 
Academy certified teacher remotely. There will be no fees for the teaching on how to 
meditate nor for any other aspect of participation in the study. Conversely, participants 
will receive no remuneration for participating in the study. 
A copy of Tao Calligraphy Greatest Love and Meditation with Tao Calligraphy instruction 
sheet will be provided to the participants electronically.  
 
Participants of the study will be required to self-meditate using the Tao Calligraphy for 
thirty minutes twice every day and to record the length of practice and responses during 
and after each practice. Besides individual meditations on their own, participants will 
attend once a week 30-minute-long group meditation practice sessions led by an 
instructor, done over the Zoom platform. 
 
Subjects in the study should continue to follow their physicians’ advice and 
recommendations during their participation in the study and should continue to receive 
any and all conventional treatment in which they have been participating prior to entry 
into this study. 

14 
 
  
Sha Research Foundation will clearly inform participants that the teaching and research 
teams are not offering any medical diagnosis, guidance, evaluation, or treatment, and 
that participation in the study is not a replacement for any conventional medical 
treatments or diagnosis. 
 
Participants are also encouraged to continue their usual spiritual practices in which they 
have been participating prior to the study. 
 
Participants of the study will be responsible for informing the research team of any and 
all of their illnesses and medications taken, but NO copies of their medical records from 
their medical practitioners or specialists will be collected besides telomere reports. 
 
The study will commence in 2023 and will last for about 2 years. The plan is to accept 
new subjects into the study at any time from its commencement in 2023 up to 2 years 
later (from April of 2023 with expected conclusion in December of 2025). 
 
A minimum of 10 and a maximum of 20 subjects may be admitted to this study during 
the study years. Subjects can withdraw from the study with no penalty at any time for 
whatever reasons they may have. 
 
 
3.0 
Enrollment Criteria (who can be in your study and who would not 
be eligible to participate in your study): 
 
Patient Population 
 
Inclusion criteria 
- 
Adult subjects / patients (age 60 years and over) we expect at 60 years people have 
already measurable shortening of telomeres 
- 
Healthy or Ill, with the exception of genetic illnesses and cancer (for which 
treatments could negatively impact telomeres) 
- 
Willingness and ability to comply with data collection requirements.  
- 
Complete submission of required documentation prior to enrollment into the 
study, including informed consent and consent to release of information. 
- 
Willingness to allow their data to be used for research purposes and published as 
deemed fit (while conforming to all applicable privacy laws) by Sha Research 
Foundation. 
- 
Willingness to practice the daily calligraphy meditations and follow the protocol. 
 
 
Exclusion criteria 
- Not meeting any of the inclusion criteria 

15 
 
- Bipolar disorders, other serious mental disorders (e.g. schizophrenia, psychosis), genetic 
illnesses (primarily affected chromosomes), and cancer (treatment could negatively 
impact telomere during research period) 
- inability to sign consent and follow instructions 
- Unwillingness to participate in data gathering 
- Unable to follow the practice regimen, including the daily calligraphy meditations 
- Pregnant or nursing. Participants who become pregnant during the study will be required 
to end their participation. (to avoid any, at current time unknown, potential negative 
effect of the study on the fetus). 
- There are no exclusion criteria placed upon potential subjects related to national origin, 
culture, ethnicity, race, sex, physical disability, sexual orientation, religion, or spiritual 
practices. 
 
 
4.0 
Sample Size: 
 
A/ Sample Size:  
The study will be conducted as a simple follow up study. It will include subjects / patients 
who are at least 60 years old at the time of the examination. 
 
The number of cases estimated was 10 (minimum) <= N <= 20 (maximum). 
 
For the future formal randomized study, we will calculate the power (based on this pilot 
study).  
 
The evaluation of the improvement in the Lengthening of Telomeres and Quality of Life 
questionnaires is done by means of the program Minitab or SPSS / PSPP using appropriate 
parametric and non-parametric tests: For statistical analysis of scores obtained from 
questionnaires we will use Anova and regression analysis, with confidence level set at 95%. 
 
 
5.0 
Recruitment and Screening Methods: 
 
Recruitment announcement with be placed at: 
Sha Research Foundation website: www.ShaResearchFoundation.com 
Sha Research Foundation Twitter and Facebook social media. 
Newspaper and magazine advertisements.  
We are submitting previously approved letter and advertisement for the study 
Pro00031067. 
 
SCREENING METHODS: 

16 
 
Member of the research team will contact candidates for the research study over the 
phone, set up zoom interviews, conduct discussions (clarification of inclusion / exclusion 
criteria), help with signing of the consent /Information, and offer instruction in the study 
design. 
Successful applicants will sign the consent and forward it to clinical / research principal 
investigators. 
 
6.0 
Research Locations: 
 
Cynthia Hamilton 
943 Beaumont Drive, North Vancouver, BC V7R 1P5 
Phone: 778 847-3617 / cynthia.hamilton@uss-lmtp.com 
 
7.0 
Multi-site Research (research that involves external collaborating 
institutions and individuals):  
 
N/A 
 
 
 
8.0 
International Research (where data collection will occur outside the 
United States and U.S. territories, including online activities) 
 
This pilot research will take place in Canada exclusively. 
 
9.0 
Procedures Involved: 
 
The study is conducted on an outpatient basis and includes: 
• A personal or telephone discussion (clarification of inclusion / exclusion criteria)  
signing of the consent /Information.  
• instruction in the study design and practicing the Tao Calligraphy-tracing 
meditation;  
• first data collection questionnaires at Time 0 
• Blood sample drawn and telomere assessment at Time 0. 
• regular Zoom practices with participants in the intervention group (or pre-
recorded meditation practices with certified teachers as a back-up) 
• Completion of the questionnaires by the participants at Time 1 (3 months)  
• Blood sample drawn and telomere assessment at Time 1 (3 months) 
• Completion of the questionnaires by the participants at Time 2 (9 months)  
• Blood sample drawn and telomere assessment at Time 2 (2 months) 
• Completion of assessments by a Study principal co-investigator  

17 
 
 
10.0 Research with Vulnerable Populations (if children are the ONLY 
vulnerable population you plan to enroll, do NOT complete this 
section -- instead fill out Appendix A) 
 
N/A 
 
 
11.0 Incomplete Disclosure or Deception: 
 
N/A 
 
 
12.0 Consent Process:  
 
Trained research team member(s) will follow SRF Consent process V1.0 20230409.  
After obtaining the name and contact of potential participant from the PI, research team 
member will connect with the potential subjects for screening using inclusion and 
exclusion criteria of the study. This will be done either in person, over the phone, or by 
Zoom meeting. If the consent is done by phone call or Zoom, subject will sign the 
consent, witnessed by a second person and mail to Clinical / Research. Participant will be 
given a study information letter for the participants, explaining the study in more detail. 
For consent form see Main ICF. 
The signed consent will be stored at Sha Research Foundation for 7 years.  
 
13.0 Waiver of Participant Signature on Consent Form: 
 
N/A 
 
 
14.0 Waivers and Alterations of Consent Information: 
 
N/A 
 
15.0 Financial Compensation: 
 
There will be no compensation of research participants.   
Participants may incur costs related to copying or faxing documents. 
Participants will receive free instruction and weekly practices with instructor. 
Participants will receive free transmission of pure energy (blessing) and electronic version 
of Tao Calligraphy. 

18 
 
Research involves minimal (no known) risk to the participants; therefore, no 
compensation will be available to the participants. 
 
 
 
16.0 Audio/Video Recording/Photography 
 
We will not be performing any video, audio, or photographic recording of subjects, but 
may request pictures of telomeres from diagnostic laboratory.  
 
 
17.0 Potential Benefits of this Research:  
 
There has been a growing number of empirical studies of Chinese calligraphic handwriting 
that resulted in improvements in visual attention and span, increased mental 
concentration, confirmation as an effective treatment for psychosomatic conditions and 
post-traumatic hyper-arousal symptoms, and exhibiting significant effects on hypertension 
and type 2 diabetes. Leisure activities occurring later in life that include calligraphy writing 
may inhibit cognitive decline.  
Our own studies clearly documented improvement in Wellbeing Scores (Rand SF-36 
questionnaire), improvement in pain scores (McGill pain questionnaire) in subjects 
suffering from chronic pain, and improvement in depression scores (BDI, PHQ9 and HAM 
questionnaires) in patients with depression. Our reports have been published and 
presented at conferences. 
 
18.0 Potential Risks to Participants: 
No risks or negative side effects of the blessings given to human participants or calligraphy 
meditation with Tao Calligraphy have been observed in our previous pilot projects and formal 
studies. 
 
 
 
 
 
 
 
19.0 Provisions to Protect Participant Privacy and Data Confidentiality: 
 
All scientific data will be stored in the computer in spreadsheet under ID code numbers 
according to Tri Council Policy Statement: Ethical Conduct for Research Involving Humans 
(2005).  

19 
 
A unique ID code number will be assigned to each participant for entry for internal tracking. 
Once the unique ID number is assigned, no personal identity information is kept with 
research spreadsheets. 
The data collected via website will have only confidential ID assigned and never any names or 
personal identifiers. As there are no names associated with participants’ responses in their 
electronic research files on the website, it would be impossible to know which subject these 
responses belong to. These data are periodically downloaded and originals are deleted from 
the website for extra layer of precaution. 
When client provides medical or personal information in paper form, these are scanned and 
forwarded to research team by email. Paper original documents are returned to participant 
who provided them. Once the email with documents is received by the research team, 
downloaded to a dedicated research external hard drive, and confirmed receipt, the emails 
with files are deleted from email inbox.   
All collected personal and medical information is stored on external hard drive that can be 
accessed only through a password-protected computer.  
There will be no hard copies of data printed out, although the results of statistical tests and 
actual papers for presentation will be in paper form. 
 
20.0 Data Monitoring Plan to Ensure the Safety of Participants: 
The collected data will remain the property of Sha Research Foundation and only members of 
the research team will have the access to the research data collected. 
 
Each participant will complete an Application Form. The signed consent forms will be emailed 
or faxed to the Foundation. Data collection will be executed at 0, 3, and 9-month intervals after 
receiving the energy transmission, for a total of 9 months. 
 
 
Research Data 
 
At the time of admission to the study, participants will provide following: 
 
1) Application that includes information of their illnesses and medications, and consent 
 
2) Laboratory assessment of Telomere Length in Peripheral Blood Leucocytes. Results of 
analysis will be stored at Repeat Diagnostics, Suite 309 – 267 West Esplanade North 
Vancouver, BC V7M 1A5 Canada for Telomere Assessment. Reports will be issued to clinical 
Principal Investigator Dr. Peter Hudoba, will be scanned and electronic copy will be stored on 
external hard drive, the paper version will be destroyed.  
 
3) Data from 
1. John Ware’s SF-36 Quality of Life questionnaire 
2. Simple Follow up Questionnaire of our design 

20 
 
 
The data collected via website will have only confidential ID assigned and never any names or 
personal identifiers. As there are no names associated with participants’ responses in their 
electronic research files on the website, it would be impossible to know which subject these 
responses belong to. These data are periodically downloaded and stored on external hard 
drive; originals are deleted from the website for extra layer of precaution. 
 
C) Data Processes 
There will a Consent Process form filled after consenting procedure. This will be reviewed. 
All applications are audited for completeness. 
The log of adverse events will be maintained. (This would also include data issues.) 
The other logs would be Protocol Deviation Log, Subject Withdrawal and Termination Log, 
Subject Visit Log spreadsheet, and Telephone Log. 
Application and consent will be scanned and stored on large (not flash drive) external drive by 
principal investigator. After he confirms receipt, any person obtaining these documents will 
delete them from their computers and from email inbox. Once principal investigator 
downloads the data from inbox to a hard drive, any email containing data will be deleted from 
his inbox.  
Research Data collected on password protected entry on the website, will have only first name 
and confidential ID as identifiers, and so even in unlikely event of breach to website, no 
research data could be attributable to a particular person. These are downloaded to external 
hard drive and periodically deleted from website. 
All data will be stored for 7 years and will be safely deleted afterwards. Any retiring hard drive 
will be not only safely erased using military protocol 3 wipe repetition and filling drive with 
Zeros (Program Avast), but also physically damaged so could not be read from anymore (in 
accordance with Canadian Medical Protective Association guidelines). 
 
 
 
21.0 Long-term Data and Specimen Storage and Sharing:  
All scientific data will be stored on external hard drive for at least 7 years under ID code 
numbers according to Tri Council Policy Statement: Ethical Conduct for Research Involving 
Humans (2005) and HIPAA. All data will be deleted after 7 years. 
No personal or attributable medical data will be shared outside of Sha Research Foundation. 
 
 
22.0 Qualifications of Research Team to Conduct the Research: 
 
Peter Hudoba De Badyn, MD, FRCS, neurosurgeon, affiliated as researcher with, and prior CEO 
of Sha Research Foundation. Previously as a Professor of Neurosurgery, University of 
Saskatchewan. Mailing address: Mt Seymour Clinic, 2nd Floor, 333 Mt Seymour Boulevard, 
North Vancouver, BC  

21 
 
 
The names of research participants removed as per Clinical Trials Gov website instructions. 
 
 
 
 
 
23.0 Publication of the Research: 
 
Authorship eligibility guidelines and any intended use of professional writers 
 
We will follow ICMJE - International standards for reporting research: 
https://phcogcommn.org/wp-content/uploads/2021/08/icmje-recommendations.pdf 
 
Authorship of the data of this study will be based on: 
1. degree of contribution to the conception or design of the study 
2. degree of creating any important intellectual content of the study 
3. degree of participation on creating final report 
4. an agreed role in the conduct of the study.   
 
The results of the study will be published on the Foundation website and in scientific meetings 
and journals. The audience is expected to be the health care, scientific, and integrative, 
complementary, and alternative medicine communities. 
 
The research data, data on the reports delivered to PI, and results of the study remain the 
intellectual property of Sha Research Foundation.  
The laboratory data and results of Telomere assessment stored at laboratory site will remain 
the property of Repeat Diagnostics, North Vancouver, BC, Canada. 
 
 
24. Budget the Research: 
 
Estimated total cost for 10 subject: $10, 983 (in Vancouver) or $14,323 (outside of 
Vancouver) plus $3,185 legal fees (that are the same for both options). 
 
Legal and Study overhead cost: 
ITEM 
COST 
INFORMATION 
IRB ethics review and 
approval 
$2,635.00 
Advarra 
Research protocol review fee 
$200.00 
VCH cost 
Research protocol set up fee 
$350.00 
VCH cost 
TOTAL OVERHEAD 
$3,185.00 
 

22 
 
 
Specimen handling and testing: 
Venipuncture Fee & 
specimen labeling/handling   
$50.00 
 
Specimen packaging* 
*See picture below for details 
$8.10-9.45 
UN3373 shipping box: 
https://www.uline.ca/ 
Biological substance category B 
FedEx priority specimen 
transportation 
$127.07 
FedEx Priority overnight 
(Toronto – Vancouver) $127.07 
– delivery next business day 
FedEx priority specimen 
transportation 
$91.05 
Within Vancouver 
Telomere length 
measurement - 2 cells 
*see attached requisitions for 
BC & Canada 
$400.00 
Repeat Diagnostics Suite 309 – 
267 West Esplanade North 
Vancouver, BC V7M 1A5 Canada 
Telomere length 
measurement - 6 cells 
$800.00 
 
TOTAL COST per 1 SUBJECT 
 
 
TOTAL COST per 1 SUBJECT 
$549.15 
 
 
$716.15 
Subject from Vancouver and 
using 2 cells Telomere length 
measurement. 
Subjects outside of Vancouver 
using 2 cells Telomere length 
measurement, additional cost is 
17$ Family doc referral, 150$ 
hematologist specialist order of 
test  
TOTAL COST per 1 SUBJECT 
 
 
 
 
949.15 
 
 
 
Subject from Vancouver and 
using 6 cells Telomere length 
measurement 
 
 
 
 
 
 
Appendices and additional documents: 
 
Appendix C: CVs 
Informed Consent Form 
Information Letter to Subjects 
Rand SF 36 Questionnaire (screenshot from website) 
Follow up form of our design (screenshot from website) 

23 
 
 
 
 
 
______________________ 
 
 __________________  
Allan Chuck 
 
           
 
    Date 
President and CEO 
Sha Research Foundation 
 
 
______________________ 
 
 __________________  
Peter Hudoba  
       
 
     
   Date 
Principal Research Investigator 
 
 
 
 
 

24 
 
 
 
i In an evaluation of minimally invasive sample collection methods for telomere length 
measurement, Oragene saliva was moderately correlated (ρ = 0.48, p = 0.002) and the 
most similar in size to buffy coat from venous blood, considered the “gold standard.” 
[17] 
ii Cross-sectional studies have revealed differences in telomeres and DNA methylation 
associated with age [22]. The latter has led to a description of a ‘molecular clock’ that is 
associated with chronological and biological age [19] [22]. 
 
iii Phenotypic age calculated using the equation provided by Liu et al. [22] [22] that uses 
chronological age and nine biomarkers, including albumin, creatinine, glucose, log (C-
reactive protein), lymphocyte percent, mean cell volume, RDW, ALKP, and white blood 
cell count. 
 
 
 
Lifestyle Intervention Meditation):  proposed biological mechanisms and study design 
implications 
From  2011 Intensive meditation training, immune cell telomerase activity, and 
psychological mediators Psychoneuroendocrinology  Volume 36, Issue 5, Pages 664-681 
Telomerase is the cellular enzyme primarily responsible for telomere length and 
maintenance. In the first study to link meditation and positive psychological change with 
telomerase activity the data suggest that increases in perceived control and decreases 
in negative affectivity contributed to an increase in telomerase activity, with 
implications for telomere length and immune cell longevity.  
Methods 
Retreat participants (n = 30) meditated for ∼6 h daily for 3 months and were compared 
with a wait-list control group (n = 30) matched for age, sex, body mass index, and prior 
meditation experience. Retreat participants received instruction in concentrative 
meditation techniques and complementary practices used to cultivate benevolent states 
of mind (Wallace, 2006). Psychological measures were assessed pre- and post-
retreat. Peripheral blood mononuclear cell samples were collected post-retreat 
(baseline/pre-retreat was not collected) for telomerase activity. Because there were 
clear, a priori hypotheses, 1-tailed significance criteria were used throughout. 
Results 
Telomerase activity was significantly greater in retreat participants than in controls at 
the end of the retreat (p < 0.05). Increases in Perceived Control, decreases in 
Neuroticism, and increases in both Mindfulness and Purpose in Life were greater in the 
retreat group (p < 0.01). Mediation analyses indicated that the effect of the retreat on 
telomerase was mediated by increased Perceived Control and decreased Neuroticism. In 
turn, changes in Perceived Control and Neuroticism were both partially mediated by 
increased Mindfulness and Purpose in Life. Additionally, increases in Purpose in Life 

25 
 
 
directly mediated the telomerase group difference, whereas increases in Mindfulness 
did not. 
[25] C. Dal Lin et al.: 2021, Rapid changes of miRNAs-20, -30, −410, −515, −134, 
and −183 and telomerase with psychological activity: A one year study on the 
relaxation response and epistemological considerations.  Journal of Traditional and 
Complementary Medicine 
Telomerase is a cellular enzyme that adds the necessary telomeric DNA (T2AG3 repeats) 
to the 3 ′-end of the telomeres, protecting their degeneration.74 The activity of this 
enzyme represents a marker of cellular aging75 and is implicated in aging-associated 
diseases.76 However, compared to telomere length, telomerase function seems to 
correlate more faithfully with stress-related psychological mechanisms.75 Stress leads 
to a decrease in its activity77 while relaxation favors its functioning,78 as our results 
appear to confirm. 
The telomerase activity (Fig. 3) appeared to increase during the RR sessions (p < 0.01 
Wilcoxon test at every time point) both in patients and in healthy volunteers. No 
significant variation was found in the CONTROLS (p > 0.05 Wilcoxon test at every time 
point). The basal values (before and after 20 min) of telomerase activity were 
comparable in all groups and were merged into a single starting basal point. Micro-RNAs 
of cardiovascular interest, involved in cell senescence and in the NF-κB network 
(miRNAs −20, −30, −410, −515, −134, and −183), and the activity of telomerase in 
peripheral blood mononuclear cells (PBMCs) were measured in the serum of patients 
with ischemic heart disease (and healthy controls) immediately before and after a 
relaxation response session (RR), three times (after the baseline), in one year of follow-
up. Results: miRNA-20 and -30 levels and PBMCs-telomerase activity increased during 
the RR while the −410 and −515 levels decreased. During the RR sessions, both miRNA-
134 and -183 decreased.  
The orientation of mental processes, either toward stress or toward relaxation, can 
impact cellular ageing through at least three main recognized pathways: the immune 
system,70 the oxidative balance,71 and the activity of telomerase.72  Although telomere 
length is implicated in cellular ageing, the evidence suggesting telomere length is a 
biomarker of ageing in humans is equivocal,73 if not invalid.57 
 
As stated, we previously assessed malondialdehyde (oxidative-stress marker) and 
galectin-3 (immune system-inflammatory marker) levels as aging58,59 and cellular 
senescence markers,60, 61, 62, 63 demonstrating a decrease in the course of 
RR.64 Numerous studies in the literature and our previous studies12,19 have shown that 
RR causes a decrease in the levels of inflammatory cytokines, stress hormones, 
inflammatory genes expression,19 and epigenetic markers of ageing.20 
 

26 
 
 
From 2017 Effect of heartfulness meditation on burnout, emotional wellness, and telomere 
length in health care professionals, Journal of Community Hospital Internal Medicine 
Perspectives, [26] 
 
ABSTRACT Background: Burnout poses significant challenges during training years in residency 
and later in the career. Meditation is a tool to treat stress-related conditions and promote 
wellness. Telomere length may be affected by burnout and stress. However, the benefits of 
meditation have not been fully demonstrated in health care professionals. Objective: We 
assessed the effects of a 12-week ‘Heartfulness Meditation’ program on burnout, emotional 
wellness, and telomere length in residents, faculty members, and nurses at a large community 
teaching hospital during the 2015–16 academic year. Methods: All subjects completed a 
baseline Maslach Burnout Inventory (MBI) and Emotional Wellness Assessment (EWA) at the 
beginning of the study. Meditators received instructions in Heartfulness Meditation. At week 12, 
subjects completed a follow up MBI and EWA scores. Salivary telomere length was measured at 
baseline and week 12. Results: Twenty-seven out of a total 155 residents (17.4%) along with 
eight faculty physicians and 12 nurses participated in the study. Thirty-five enrolled as 
meditators and 12 as controls. At 12 weeks, the meditators had statistically significant 
improvement in all measures of burnout and in nearly all attributes of EWA. Controls showed no 
statistically significant changes in either burnout or emotional wellness scores. Relative 
telomere length increased with statistical significance in a younger subset of meditators. 
Conclusion: Our results indicate that meditation offers an accessible and efficient method by 
which physician and nurse burnout can be ameliorated and wellness can be enhanced. The 
increased telomere length is an interesting finding but needs to be confirmed with further 
research. Abbreviations: EWA: Emotional wellness assessment; MBI: Maslach burnout 
inventory; EE: Emotional exhaustion; DP: Depersonalization. 
 
From: Exploring Epigenetic Age in Response to Intensive Relaxing Training: A Pilot Study to 
Slow Down Biological Age    
Abstract  DNA methylation (DNAm) is an emerging estimator of biological aging, i.e., the often-
defined “epigenetic clock”, with a unique accuracy for chronological age estimation (DNAmAge). 
In this pilot longitudinal study, we examine the hypothesis that intensive relaxing training of 60 
days in (20) patients after myocardial infarction and in (10) healthy subjects may influence 
leucocyte DNAmAge by turning back the epigenetic clock. Moreover, we compare DNAmAge 
with another mechanism of biological age, leucocyte telomere length (LTL) and telomerase. 
DNAmAge is reduced after training in healthy subjects (p = 0.053), but not in patients. LTL is 
preserved after intervention in healthy subjects, while it continues to decrease in patients (p = 
0.051). The conventional negative correlation between LTL and chronological age becomes 
positive after training in both patients (p < 0.01) and healthy subjects (p < 0.05). In our subjects, 
DNAmAge is not associated with LTL. Our findings would suggest that intensive relaxing 
practices influence different aging molecular mechanisms, i.e., DNAmAge and LTL, with a 
rejuvenating effect. Our study reveals that DNAmAge may represent an accurate tool to 
measure the effectiveness of lifestyle-based interventions in the prevention of age-related 
diseases. 
 

27 
 
 
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28 
 
 
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