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INFORMED CONSENT
Informed consent was obtained from all individual participants included in the study.
PMC10315327
TRIAL REGISTRATION
Iranian Registry of Clinical Trial (registered prospectively: IRCT20191112045424N1; available at:
PMC10315327
ACKNOWLEDGMENTS
We are tankful to all study participants. We would like to thank the Clinical Research Development Unit, Imam Reza Hospital, Mashhad University of Medical Sciences, for their assistance in this manuscript.
PMC10315327
DATA AVAILABILITY STATEMENT
The datasets used and analyzed during the current study are available from the corresponding author on reasonable request.
PMC10315327
REFERENCES
PMC10315327
Objective
NONALCOHOLIC FATTY LIVER DISEASE, TYPE 2 DIABETES
Academic Editor: Ferdinando Carlo Sasso The aim of study was to evaluate the effect and safety of pioglitazone-metformin combined treatment in the newly diagnosed type 2 diabetes patients with nonalcoholic fatty liver disease.
PMC10250100
Methods
nonalcoholic fatty liver disease
NONALCOHOLIC FATTY LIVER DISEASE, TYPE 2 DIABETES
A total of 120 newly diagnosed type 2 diabetes patients with nonalcoholic fatty liver disease from 8 centers were randomly divided into the control group (metformin hydrochloride) and the test group (pioglitazone hydrochloride and metformin hydrochloride).
PMC10250100
Results
fatty liver
FATTY LIVER
Compared to the control group, after treatment, the proportion of people with mild and moderate fatty liver increased, and the proportion of people with severe fatty liver decreased, and this change was more obvious in the population with moderate and severe fatty liver. The level of
PMC10250100
Conclusion
diabetic
ADVERSE EVENTS, NONALCOHOLIC FATTY LIVER DISEASE
Combined treatment with pioglitazone-metformin can effectively reduce liver fat content and gamma-GT level in newly diagnosed diabetic patients with nonalcoholic fatty liver disease, and adverse events do not increase compared with the control group, showing good safety and tolerance. This trial is registered with ClinicalTrials.gov
PMC10250100
1. Introduction
NAFLD, fatty liver, weight gain, hypoglycemic, type 2 diabetes, diabetes
ADVERSE EVENTS, FATTY LIVER, CHRONIC DISEASES, INSULIN RESISTANCE, TYPE 2 DIABETES, DIABETES
As one of the most common chronic diseases, the prevalence rate of diabetes in China has skyrocketed, and the estimated prevalence of diabetes in Chinese adults is 11.6%, with type 2 diabetes predominating [Clinically, NAFLD is currently treated with multiple drugs, one of which is insulin sensitizer. Many literature reports that insulin sensitizers can effectively improve NAFLD [Therefore, the purpose of this study is to observe the therapeutic effect of pioglitazone-metformin combined treatment in type 2 diabetes patients with NAFLD. While using pioglitazone to improve the patient's liver histological performance, additional metformin can offset the risk of weight gain caused by pioglitazone. In addition, both can improve the patient's liver enzyme levels and insulin resistance, thereby increasing patient compliance, and bring obvious benefits to patients with type 2 diabetes and fatty liver.The hypoglycemic effect of pioglitazone hydrochloride and metformin hydrochloride tablets (15 mg pioglitazone/500 mg metformin) has been fully verified and affirmed. In this study, ultrasound liver fat content and liver enzyme levels were used as primary endpoint to evaluate the efficacy of NAFLD. In this multicenter clinical trial, we used a randomized, double-blind, double-simulated method, with metformin as the control group and ultrasound liver fat content and liver enzyme levels as the primary endpoints to evaluate the effect of combined treatment of metformin and pioglitazone in type 2 diabetes patients with NAFLD; the risk of adverse events is used to evaluate the safety of combined treatment of metformin and pioglitazone.
PMC10250100
2. Materials and Methods
PMC10250100
2.1. Study Design and Participants
This study was a randomized, double-blind, double-simulation, and positive drug control multicenter clinical trial (ClinicalTrials.gov registration number: Inclusion criteria are as follows: according to WHO criteria, age 18-70 years old, BMI between 21 and 35 kg/m
PMC10250100
2.2. Randomization and Masking
LIVER
Through a computer-generated centralized management program, all participants were randomized by a stratified computed randomization procedure to account for age, sex, and BMI to test group or control group at a ratio of 1 : 1 and were masked to the treatment assignment. The test group received pioglitazone-metformin combined treatment, and the control group received metformin treatment and matching placebo treatment. The electronic master randomization list was only accessible to the assigned randomization list managers, and study sites received sealed opaque envelopes for unblinding in cases of emergency. Enrollment was performed at the respective site. Randomization and assignment to the double-blind study drug were done by central pharmacy personnel, who had access to the computer-generated randomization scheme. The appearance, color, and smell of all simulated tablets are the same as the corresponding drugs. Liver fat content testing in all test centers is performed by fixed personnel, and all testing personnel have been uniformly trained to ensure the same measurement quality.
PMC10250100
2.3. Procedures
thirst, gastrointestinal tolerance, overdose, hypoglycemia, weight loss
HYPOGLYCEMIA, CONCOMITANT DISEASE
In order to improve gastrointestinal tolerance, all patients in the group received a weekly dose titration. The initial dose of the test group was once a day, 500 mg metformin and 15 mg pioglitazone (both from Hangzhou Zhongmei Huadong Pharma Ceutical Co., Ltd.) were taken before breakfast or dinner, and after 1 week of treatment, the dose was adjusted to twice a day, before breakfast and dinner. The initial dose of the control group was 500 mg metformin, once a day, before breakfast or dinner, and after 1 week of treatment, the dose was adjusted to twice a day, before breakfast and dinner. Patients were visited at 2, 4, 8, 12, 16, 20, and 24 weekends after treatment to evaluate the effectiveness and safety of the trial drug.Participants will withdraw from the experiment when the following situations occur: severe hypoglycemia requires the assistance of others; fasting blood glucose < 2.8 mmol/L or two random blood glucose < 3.9 mmol/L caused by overdose; in the absence of other concomitant diseases, frequent urination, thirst, weight loss, or other aggravation due to high blood sugar; and asymptomatic FPG confirmed by two measurements > 13.3 mmol/L.
PMC10250100
2.4. Quantitative Ultrasound for Liver Steatosis
In our study, we mainly used controlled attenuation parameter (CAP) for liver fat quantification, which is best studied and clinically available technique for liver fat quantification, with the first clinical studies dating back to 2010 [
PMC10250100
2.5. Outcomes
fasting blood glucose, hypoglycemic, postprandial blood glucose
EVENTS, SECONDARY, INSULIN RESISTANCE
The primary end points are liver fat content (ultrasonic quantitative determination of liver fat content), ALT, AST, gamma-GT levels, and insulin resistance index. The secondary endpoints are HbA1c, intravenous fasting blood glucose, 2 h postprandial blood glucose, total cholesterol, triglyceride, HDL cholesterol, LDL cholesterol, waist circumference, and body weight. Safety indicators such as medication compliance, vital signs, hypoglycemic events, nonhypoglycemic events, blood routine, urine routine, and electrocardiogram were analyzed.
PMC10250100
2.6. Statistical Analysis
The statistical analysis was performed using SAS (9.1.3) statistical analysis software. The
PMC10250100
3. Results
PMC10250100
3.1. Baseline Characteristics
NAFLD
TYPE 2 DIABETES
This study enrolled a total of 120 type 2 diabetes patients with NAFLD from 8 hospitals in Shaanxi Province. As shown in the Consolidated Standards of Reporting Trials (CONSORT) diagram in
PMC10250100
3.2. Primary Endpoint Measurement
After 24 weeks of treatment, 8 (13%) of the 60 patients in the test group had substantial improvement in degree of liver fat content, and the degree of liver fat content decreased from moderate and severe grade to mild grade. In contrast, 4 of 60 patients (6%) in the metformin group saw their liver fat content drop from moderate or severe to mild. Statistical analysis showed that the differences in the test group before and after treatment and compared with the metformin group were statistically significant. The number of patients with moderate or lower liver fat content in the pioglitazone-metformin combined treatment group increased from 46 (76.7%) to 54 (90.0%) and the metformin group increased from 43 (71.7%) to 47 (78.3%). The changes before and after treatment in the pioglitazone-metformin combined treatment group were statistically significant (Compared with the metformin group, pioglitazone-metformin combined treatment was associated with a significant decrease in
PMC10250100
3.3. Secondary Endpoint Measurement
After 24 weeks of treatment, the HbA1c level of the pioglitazone-metformin combined treatment group and the metformin treatment group decreased significantly (pioglitazone-metformin combined treatment: 1.78% (95% CI: 1.53-2.04) and metformin treatment: 1.71% (95% CI: 1.46-1.96)), but there was no significant difference between the two groups (There is no statistically significant reduction in BMI, and waist circumference was observed in the pioglitazone-metformin combined treatment group after 24-week treatment compared to the metformin. Moreover, there was no significant change in the pioglitazone-metformin combined treatment group and the metformin group when compared with the baseline level (
PMC10250100
3.4. Safety Evaluation
In this study, the compliance of pioglitazone-metformin combined treatment group was 95.79%, and that of the metformin group was 95.10%. As shown in
PMC10250100
4. Discussion
NAFLD, occult blood, diarrhea, fatty liver, adverse cardiovascular events, angina pectoris, liver cirrhosis, ALD, NAFL, hypoglycemic, metabolic stress liver injury, hypertension
ADVERSE REACTIONS, ALCOHOLIC LIVER DISEASE, HYPERTENSION, ADVERSE EVENTS, NONALCOHOLIC STEATOHEPATITIS (NASH), DISEASE, NONALCOHOLIC FATTY LIVER, FATTY LIVER, LIVER CIRRHOSIS, INSULIN RESISTANCE, HEPATOCELLULAR CARCINOMA, NONALCOHOLIC FATTY LIVER DISEASE, ALD, TYPE 2 DIABETES
Nonalcoholic fatty liver disease is a metabolic stress liver injury closely related to insulin resistance and genetic susceptibility. Except that the patient has no history of excessive drinking, its pathological changes are similar to alcoholic liver disease (ALD). The disease spectrum includes nonalcoholic fatty liver (NAFL), nonalcoholic steatohepatitis (NASH), and related liver cirrhosis and hepatocellular carcinoma [In this double-blind, randomized, placebo-controlled trial, pioglitazone-metformin combined treatment met the predefined primary endpoint and can significantly reduce the liver fat content of newly diagnosed type 2 diabetes with moderate or severe fatty liver patients. In addition, after 24 weeks of treatment, the patient's No statistically significant changes were observed in patients with mild liver content; this is consistent with previous studies showing that insulin sensitizers can effectively improve NAFLD [In terms of safety, the addition of metformin did not significantly increase the risk of adverse events. The pioglitazone-metformin combination group had the same risk of adverse events as the metformin group, and they were well tolerated. Drug-related adverse reactions were mainly diarrhea, occult blood in the urine, and insufficient blood supply to the myocardium. These adverse events were, however, mainly transient and mild-to-moderate severity. A total of 2 serious adverse events occurred in the two groups, including hypertension and angina pectoris, with mild-to-moderate severity. Both cases were considered to be unrelated to the study drug. However, whether pioglitazone affects the cardiovascular system and the occurrence of adverse cardiovascular events remains to be confirmed by further increasing the number of studies. NAFLD is a recent research hotspot, and there is still no specific guideline for the treatment plan of NAFLD patients. Although many hypoglycemic drugs have been tested in NAFLD patients, there are currently no approved drugs for the treatment of NAFLD [
PMC10250100
5. Limitation
NAFLD
Although our research is a randomized, double-blinded, double-simulated multicenter study, but our research still has some limitations. First of all, our study did not incorporate the gold standard of liver biopsy into our main results because of the expected early stages of NAFLD in these patients and the short duration of intervention, which weakened the reliability of our results to a certain extent. Moreover, this study did not use multiple imputation to account for missing values but performed maximum likelihood methods for the primary end point. In addition, 24 patients were lost during follow-up (20%) in our study. This could be a huge missing and may represent a bias to the study. Finally, due to our inclusion of patients and the limited duration of intervention, long-term, large-scale placebo-controlled, rigorous randomized controlled studies are still required to confirm our results.
PMC10250100
6. Conclusion
NAFLD, fatty liver
FATTY LIVER, TYPE 2 DIABETES
In conclusion, the results of this study showed that in the observation period of 24 weeks, in newly diagnosed type 2 diabetes patients with NAFLD, the pioglitazone-metformin combined treatment can effectively reduce liver fat content, especially in patients with moderate to severe fatty liver and
PMC10250100
Acknowledgments
We thank all the participants, investigators, and trial-site staff who were involved in the conduct of the study. We also thank Hangzhou Zhongmei Huadong Pharma Ceutical Co., Ltd. for providing free experimental drugs and placebos. This study was funded by the Xijing Hospital.
PMC10250100
Data Availability
The original contributions presented in the study are included in the article. Further inquiries can be directed to the corresponding author.
PMC10250100
Ethical Approval
The research protocol was approved by the Ethics Committee of Xijing Hospital of Air Force Medical University (ClinicalTrials.gov registration number:
PMC10250100
Consent
Written informed consent was obtained from all the participants after a full explanation of the study.
PMC10250100
Disclosure
This trial represents independent academic research funded by Xijing Hospital. The trial drug and placebo were provided free of charge by Hangzhou Zhongmei Huadong Pharma Ceutical Co., Ltd., which did not participate in the design of the experiment, collection, analysis, and interpretation of data, report writing, and the decision to submit.
PMC10250100
Conflicts of Interest
All authors participating in this study reported no conflicts of interest.
PMC10250100
Authors' Contributions
±, fatty liver
ADVERSE EVENTS, FATTY LIVER
F.J.F and J.Q.H participated in the conception and design of this research, explained the data, and critically revised the manuscript. All authors have read and approved the final version. All authors participated in data collection, collation, and manuscript writing. All authors are guarantors of this work and are responsible for the completeness of the data and the accuracy of data analysis.Consolidated Standards of Reporting Trials (CONSORT) diagram of participant flow through the study.(a) The number of patients with mild fatty liver at baseline, 12 weeks, and 24 weeks. (b) The number of patients with moderate and severe fatty liver at baseline, 12 weeks, and 24 weeks. (a) Plasma Baseline characteristics of study population.Abbreviations: SBP: systolic blood pressure; DBP: diastolic blood pressure. Data are Body weight, waist circumference, and liver biological indicators at different times.Note: data are mean ± standard deviation. Incidence of adverse events in each group. Drug-related adverse events in each group.
PMC10250100
Supplementary Information
lymphopenia
LYMPHOPENIA, MYOCARDIAL INFARCTION (MI)
Myocardial infarction (MI) accelerates immune ageing characterised by lymphopenia, expansion of terminally differentiated CD8The online version contains supplementary material available at 10.1007/s11357-023-00794-6.
PMC10122201
Keywords
PMC10122201
Introduction
death
MYOCARDIAL INFARCTION (MI), ACUTE CORONARY SYNDROME, PROLIFERATION, INFLAMMATION, VIRAL INFECTION, EVENTS
Despite modern advances in therapy, myocardial infarction (MI) still carries a substantial risk of death or recurrent cardiovascular events [Immunosenescence is mainly linked to thymic involution and changes in cellular immunity as a response to pathogens, such as recurrent viral infections, throughout life [Lymphocyte proliferation can be enhanced in vitro by activating telomerase, a reverse transcriptase that typically counteracts telomere shortening at the end of chromosomes [We present the results from the Telomerase ACTivator to reverse Immunosenescence in Acute Coronary Syndrome (TACTIC) trial—the first randomized controlled trial investigating whether treatment with TA-65 for 12 months after MI reduces immune ageing and inflammation.
PMC10122201
Methods
PMC10122201
Trial design and oversight
coronary heart disease
CORONARY HEART DISEASE
The TACTIC trial was a single-centre, randomized, double-blind, parallel-group, placebo-controlled phase IIa pilot trial comparing TA-65 with placebo in 90 participants with coronary heart disease who were diagnosed with myocardial infarction in the 6 months prior to enrolment. A total of 90 patients were randomized to either the TA-65 group (
PMC10122201
Trial population
comorbidity, substance addiction, obstructive coronary artery disease, malignancy, NSTEMI, epicardial vessel stenosis, arrhythmias, immunological dysfunction, hepatitis
ACUTE MYOCARDIAL INFARCTION, UNCONTROLLED HYPERTENSION, MYOCARDIAL INFARCTION, BLOOD, EVENT, DISEASE, CARDIOGENIC SHOCK, ARRHYTHMIAS, HEPATITIS, INSULIN-DEPENDENT DIABETES MELLITUS
Patients were eligible for the trial if they a) were aged 65 years or over with an index presentation of an acute myocardial infarction (either STEMI or NSTEMI) within the previous 6 months, b) had successfully completed revascularisation or were being managed medically following myocardial infarction, c) had evidence of obstructive coronary artery disease on invasive coronary angiography (at least one major epicardial vessel stenosis ≥ 70%), and d) were more than 24 h after presentation with the index event—patients were eligible the following day post percutaneous coronary intervention (PCI) or 3 months after CABG. Exclusion criteria included any condition associated with immunological dysfunction (acute or chronic inflammatory or neoplastic co-existing disease, known positive serology for HIV, or hepatitis); clinical instability (arrhythmias, cardiogenic shock, unconscious); severe, uncontrolled hypertension (Blood Pressure > 170/110 mmHg, or ambulatory BP of 150/95 mmHg); severe comorbidity likely to impact on outcome over the next 2 years, immunosuppressants, known malignancy, patients already taking a nutritional supplement derived from the roots of the Astragalus species, known current or previous substance addiction and insulin-dependent diabetes mellitus.
PMC10122201
Randomization and blinding
NSTEMI
Randomisation was performed using a minimisation scheme to ensure patients randomised to each group were comparable at baseline. The minimisation scheme accounted for gender (male or female), type of MI (STEMI or NSTEMI) and proportion of terminally differentiated effector memory CD8
PMC10122201
Outcome measures
The primary outcome measure was the proportion of CD8
PMC10122201
Flow-cytometric assays
Two different assays were used for flow cytometric analysis of leucocytes. All measurements were performed on fresh blood within 4 h of collection. There was no significant amount of non-viable cells (< 1%) with this approach. The TruCount assay (containing CD45, CD3, CD4, CD8, CD19, CD16, CD56 fluorochrome-labelled monoclonal antibodies) provided information on the absolute concentration of major leukocyte populations (Fig. Gating of main leukocyte populations with TruCount assay. Flow cytometric analysis of a representative patient from the TACTIC trial is shown. At first, lymphocytes, monocytes, neutrophils, and beads were gated in SSC-Area against the leukocyte marker CD45. Lymphocytes were sequentially gated into CD3
PMC10122201
Telomerase activity
LYSED
Nuclear telomerase activity in peripheral blood mononuclear cells (PBMCs) was measured using the Telomerase Repeated Activation Protocol (TRAP) – quantitative polymerase chain reaction (qPCR) assay. Within 24 h of collection, whole blood was centrifuged to isolate PBMCs, which were cryopreserved at − 80 C. The cells were subsequently thawed, lysed, and analysed using a validated TRAP-qPCR protocol (Supplementary Tables
PMC10122201
Oxidative stress
OXIDATIVE STRESS
Oxidative stress was measured using the TBARS colorimetric assay. This assay is described in detail in the
PMC10122201
Endothelial function
hyperaemia
HYPERAEMIA, PAT
Endothelial function was evaluated using the EndoPAT device (Itamar Medical Ltd, Caesarea, Israel). This non-invasive operator-independent system consists of finger probes that measures peripheral arterial tone (PAT) for 5 min at rest and during reactive hyperaemia induced by 5-min forearm cuff occlusion. The reactive hyperaemia index (RHI) is the post- to pre- occlusion PAT signal ratio in the occluded arm relative to the same ratio in the control arm. RHI was measured at baseline, 6 months and 12 months (Supplementary Figure
PMC10122201
Echocardiography
Transthoracic echocardiography (TTE) was performed at baseline and 12 months to evaluate left ventricular function including global longitudinal strain (
PMC10122201
Adverse events
ADVERSE REACTIONS, ADVERSE EVENT, EVENT, ADVERSE EVENTS
Adverse events (AEs), adverse reactions (ARs), serious adverse events (SAEs), and serious adverse reactions (SARs) were assessed by a medical practitioner for severity and causality. The severity of each was classified as follows: grade 1—minor event not requiring medical intervention, grade 2—an event which is symptomatic and may require medical attention, grade 3—a significant event which requires medical intervention and may require hospitalisation, grade 4—an event that requires urgent medical intervention and is potentially life-threatening, grade 5—death.
PMC10122201
Statistical analysis
Continuous variables were reported by number, mean ± SD, median (IQR) or minimum and maximum, whereas number and percentages summarised categorical variables. The proportion of CD8
PMC10122201
Results
PMC10122201
Trial progress
All patients were recruited at The James Cook University Hospital, Middlesbrough, UK, between 21Consolidated Standards of Reporting Trials (CONSORT) diagram of patient enrolment. The diagram shows the number of patients who met the inclusion or exclusion criteria and their distribution among the two study groups. TA-65 and placebo were administered as 8-mg doses twice daily for 12 months
PMC10122201
Effect on telomerase activity and oxidative stress
Nuclear telomerase activity did not change significantly between baseline and 12 months in either group. The estimated change after 12 months in telomerase activity was + 22.9 (95% CI − 91.8–136.0) in the placebo group, and + 18.5 (95% CI: − 89.4–125.9) in the TA-65 group (Table
PMC10122201
Effect on endothelial function and cardiac function
There were no significant treatment effects of TA-65 on reactive hyperaemic index after 6 or 12 months compared to baseline (Table
PMC10122201
Discussion
MYOCARDIAL INFARCTION
In this novel clinical trial investigating TA-65 versus placebo in patients aged 65 years and over following myocardial infarction, we established four main results: there was no significant change in senescence-like CD8
PMC10122201
Reversal of lymphopenia under TA-65
lymphopenia, NSTEMI
LYMPHOPENIA
We demonstrated that TA-65 led to a reversal of relative lymphopenia. This was notable in NSTEMI but not STEMI patients. In STEMI, baseline lymphocytes are elevated between 24 and 72 h post reperfusion [
PMC10122201
Mitochondrial dysfunction in T cells as a driver of senescence and inflammageing
mitochondrial dysfunction
MITOCHONDRIAL DYSFUNCTION
Senescence can be driven directly by mitochondrial dysfunction [
PMC10122201
Mitochondrial telomerase activation as a potential mechanism of TA-65
Although there is no direct evidence to suggest that mitochondrial Tert protects against senescence induction, global TERT activation inhibits senescence [
PMC10122201
Inflammation as a residual risk post ACS
hypercholesterolemia, myocardial infarction, TA-65, impaired innate immunity, inflammation, infections, sepsis, platelet aggregation, coronary artery disease
HYPERCHOLESTEROLEMIA, MYOCARDIAL INFARCTION, ADVERSE EVENTS, INFLAMMATION, INFECTIONS, EVENTS, SEPSIS, CORONARY ARTERY DISEASE
Secondary prevention in patients who have undergone revascularisation after myocardial infarction aims to reduce their residual risk, predominantly by targeting hypercholesterolemia (with cholesterol-lowering agents) and platelet aggregation (with antiplatelet therapy). Recently, inflammation, as quantified by hsCRP, has been added to these determinants of residual risk. The CANTOS trial demonstrated that reducing inflammation in patients with coronary artery disease and elevated hsCRP (> 2 mg/l) reduced the rate of major adverse cardiovascular events [The major limitation of canakinumab in the CANTOS trial was the effect of IL-1β blockade on immunity. Patients in the treatment group had more infections, and a higher risk of dying from sepsis. However, our data shows that patients treated with TA-65 have no significant change in myeloid cells (monocytes and neutrophils) compared to placebo, suggesting they do not have impaired innate immunity. Indeed, their adaptive immunity appears to be enhanced, as indicated by an increase in the number of all lymphocyte subsets. An increase in lymphocyte count from 1600 to 1900 cells/µL (similar to what we have observed with TA-65 treatment in this study) correlates with a 50% reduction in the hazard ratio for both all-cause mortality and cardiovascular mortality in the large JAMA cohort study of 50,000 healthy Americans [In summary, the telomerase activator TA-65 appears to enhance adaptive immune cell numbers in the peripheral blood without negatively affecting innate immunity. We observed a trend towards reduced inflammation in patients following MI, and we hypothesise based on work by other groups that improved mitochondrial function in T lymphocytes leads to a reduction in cytokine generation and hence reduced inflammation. Clinically, there was no evidence for an increase in adverse events with TA-65 – in fact there were significantly fewer adverse events in the TA-65 group.
PMC10122201
Limitations
Our trial did not have an age matched healthy control. It is not clear whether TA-65 preserves lymphocyte count for the age group or accelerates its return to normal after MI. While a reduction in hsCRP is demonstrated, measurement of systemic cytokine levels is needed to elucidate the impact of TA-65 on SASP. We used the Becton Dickinson 6-colour Trucount assay which has premixed antibodies that have been tested vigorously in clinical settings. It only comes with CD16, and not CD14. This limits our interpretation of monocyte subtypes as CD16 alone is not a fully reliable marker to distinguish intermediate, non-classical and classical monocytes. The mechanism of TA-65 remains unexplained. We plan to explore potential mechanisms in future studies by measuring telomere DNA length and mitochondrial functional assays in stored PMBC samples. Finally, the effects of TA-65 on lymphocyte count elevation and hsCRP reduction warrant an adequately powered study.
PMC10122201
Conclusion
myocardial infarction, TA-65
MYOCARDIAL INFARCTION
Our pilot trial suggests that activation of telomerase could be a promising new drug target for patients following MI. Given the significant findings in lymphocyte increase, the trend in hsCRP reduction, as well as the extremely favourable safety profile, we propose as a next step a larger phase II clinical trial with multiple centres. We would set out to confirm whether one-year treatment with TA-65 following myocardial infarction can reduce hsCRP, shifting patients from a high to a low risk inflammatory profile.
PMC10122201
Supplementary Information
Below is the link to the electronic supplementary material.Supplementary file1 (DOCX 11827 KB)
PMC10122201
Acknowledgements
HEART
The trial was funded by TA-Sciences Inc (New York, US) as an investigator-led grant to I.S.BK is supported by a British Heart Foundation personal chair (CH//13/2/30154). K. Stellos is supported by grants from the European Research Council (ERC) under the European Union's Horizon 2020 research and innovation programme (MODVASC, grant agreement No 759248) and the German Research Foundation DFG (SFB834 B12, project number 75732319).
PMC10122201
Data availability
Data are available upon request and following approval by the trial steering committee.
PMC10122201
Declarations
PMC10122201
Conflict of interest
The authors declare no competing interests.
PMC10122201
References
PMC10122201
Background
mental illness
Some children of parents with mental illness need support. This study aimed to develop and test the effectiveness of an e-learning program for training elementary schoolteachers to support children of parents with mental illness.
PMC10024023
Methods
mental illness
The program, which included a 30-min video-based e-learning program, aimed to help schoolteachers gain basic knowledge about mental illness and children of parents with mental illness, recognize children in need of support, and gain confidence in supporting them. A school-based cluster randomized controlled trial was conducted, and the schools were randomly divided into intervention and control groups. The teachers at these schools signed up for the program and participated individually. The outcome measures for the schoolteachers were evaluated at three time points: baseline (T1), post (T2), and one month later (T3). Along with the Sense of Coping Difficulty subscale (primary outcome measure), the following self-developed outcome measures were used: actual behaviors and attitude toward supporting children, knowledge, and self-assessment of program goals achievement. The Sense of Coping Difficulty subscale results at T3 were compared between the groups. Effectiveness over time was assessed for all the outcome measures. The interaction between baseline and intervention effects on the Sense of Coping Difficulty subscale was analyzed. As a part of the process evaluation, open-ended text responses were analyzed qualitatively.
PMC10024023
Results
Baseline responses were collected from 87 participants in the intervention group and 84 in the control group. The total score of the Sense of Coping Difficulty subscale at T3 was significantly lower in the intervention group than in the control group (
PMC10024023
Conclusions
mental illness
The program was effective for schoolteachers in supporting children of parents with mental illness.
PMC10024023
Trial registration
UMIN000045483; 14/09/2021.
PMC10024023
Keywords
PMC10024023
Background
mental illness
Mental illness is an important public health issue affecting many people, with one in five Japanese individuals reportedly being affected during their lifetime [In Japan, it has only been about 15 years since the existence of children raised by parents with mental illness became known to society [Even if the children themselves are caring for their parents or struggling with family matters, they are often reluctant to seek help because of stigma [Programs for improving schoolteachers’ mental health literacy have already been developed; many of these have induced improvements in schoolteachers’ knowledge, attitudes, and confidence in supporting the mental illness of students [In Japan, 50-min video material has been developed for schoolteachers to improve their mental health literacy [This program aimed to help schoolteachers gain basic knowledge about mental illness and children of parents with mental illness, recognize children in need of support, and gain confidence in supporting them. In mental health literacy education aimed at schoolteachers, their knowledge regarding mental illness and confidence in supporting students in need of support often indicate a program’s effectiveness [
PMC10024023
Methods
PMC10024023
Design
A school-based cluster randomized controlled trial (RCT) was conducted to test the effectiveness of the program. Contamination of the information provided across groups within the same school may have occurred. Therefore, the RCTs of schoolteachers could use schools as clusters [The program, which was offered online on an individual basis, could only be watched by the intervention group. The outcome measures were evaluated using a self-administered web survey by comparing the two groups at three time points: baseline (T1), post (T2), and one month later (T3). Ethical considerations allowed the control group to participate in the same program after the final T3 response. No major changes were made to the methods after trial commencement. The trial findings were reported in a subsequent publication in accordance with the Consolidated Standards of Reporting Trials (CONSORT).
PMC10024023
Study participants
Requests for research participation were sent to 322 public elementary schools in three Japanese prefectures. Only schools that were willing to cooperate were included in the study.A maximum of 10 schoolteachers participated from each school. The inclusion criterion was full-time schoolteachers, and the exclusion criteria were difficulty in understanding Japanese and being unable to conduct web surveys or operate video watching. The sample size was determined with reference to 50-min video programs for improving schoolteachers’ mental health literacy in Japan [
PMC10024023
Program development
mental illness
DISORDERS
The program was called the WATASHI-KOKO (“We are here” in Japanese) PROGRAM. It involved a 30-min video-based e-learning program for elementary schoolteachers, which was designed to help them support children of parents with mental illness. It aimed to help schoolteachers gain basic knowledge about mental illness and children of parents with mental illness (Purpose 1), recognize children in need of support (Purpose 2), and gain confidence in supporting them (Purpose 3).The program was created with the support of “KODOMO-PEER” (“children’s companions” in Japanese), which is the largest self-help group for adult children of parents with mental illness in Japan. The program content was created based on a childhood needs survey that received responses from adult children of parents with mental illness [To gain basic knowledge about mental illness (Purpose 1), the program content was created with reference to previous research on a mental health literacy education program for schoolteachers [Components of the programTo gain basic knowledge about children of parents with mental illness (Purpose 1), the program content was created based on a survey that received responses from adult children of parents with mental illness [To help schoolteachers recognize children in need of support (Purpose 2), the program content was developed based on the survey that received responses from adult children of parents with mental illness [To help schoolteachers gain confidence in supporting children in need of support (Purpose 3), the program content was developed based on the survey [Theoretically, the program content was also based on a four-tiered approach for delivering professional development programs related to parental mental disorders [In accordance with the learning process, the content structure of the program was based on the Transtheoretical Model (TTM) of health behavior change by Prochaska et al. [Gagné’s instruction is an effective process when teaching others [The specifics of the program are shown in Table
PMC10024023
Outcome measures
The outcomes were measured at the individual level. The questionnaire was developed by multidisciplinary experts who were research members of this project. The pre-test was conducted among 18 elementary schoolteachers. After watching the video of the program, they were asked to answer the questionnaire and also asked to indicate any points where they had found it difficult to understand sentences or words. Consequently, a few points were raised by the schoolteachers, and some of the wording was corrected.
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Confidence in supporting children
The Sense of Coping Difficulty subscale was used to assess confidence in supporting children (Purpose 3 of the program). The Sense of Coping Difficulty/Possibility Scale refers to analogous concepts of self-efficacy, which is the expectation that professionals can deal with difficult situations [
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Actual behaviors and attitude
Three self-developed questions were used at T1 and T3 in order to assess the effects of program Purposes 2 and 3 on behaviors and attitudes. The program introduced the life situations of children in need of support and highlighted certain signs to look out for when identifying such children. The first question asked schoolteachers whether, over the past month, they had looked at children with the perspective that they could be children in need of support. The second question asked schoolteachers whether, over the past month, they had recognized the presence of a child(ren) in need of support. The program recommended that schoolteachers who had recognized a child in need of support first consult with other teachers in the school instead of worrying about the situation by themselves. The third question asked schoolteachers whether, over the past month, they had consulted other schoolteachers within the school about a child(ren) in need of support. The schoolteachers responded to these questions using “yes” or “no” options.
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Knowledge
mental illness
Knowledge questions, with responses that could be correct or incorrect, were administered to the schoolteachers; they were developed in order to assess program Purpose 1 (gaining basic knowledge about mental illness and children of parents with mental illness). The five items related to parental mental illness were lifetime prevalence of mental illness, types of mental illness, age of onset of mental illness, difficulties of living with people with mental illness, and mental health and welfare life support services. The five questions related to children were about the percentage of children with parents having mental illness, children's own awareness, counseling, impact on health, and the common roles of young carers. Some of the items were based on previous research on mental health literacy education [
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Program goals achievement
mental illness, illness
The degree to which the participants had achieved the eight items representing the program goals was measured using a seven-point scale ranging from “not at all” (1 point) to “very well” (7 points). The following goals were set as an assessment for achieving program Purpose 1: 1) be able to describe the impact of parental mental illness on children’s lives, 2) be able to describe children’s feelings, and 3) be able to describe the significance of school for children. The following goals were set as an assessment for achieving program Purpose 2: 4) be able to recognize children in need of support. The following goals were set as an assessment for achieving program Purpose 3: 5) be able to describe how to support children, 6) be able to act appropriately toward children, and 7) be able to act appropriately in school. Professionals’ negative attitudes regarding parenting with mental illness can hamper support provision [
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Process and feasibility evaluation measures
The process and feasibility evaluation measures were as follows: program satisfaction (very satisfied, somewhat satisfied, not very satisfied, not satisfied at all); whether the participants would recommend the program to other schoolteachers (would recommend, would not recommend); length of the program (long, just right, short); what the participants learned was new (yes/no); if yes, what was impressive (open-ended text response); whether their way of thinking had changed (changed/not changed); if so, how it had changed (open-ended text response); and how they planned to act (open-ended text response).
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Basic characteristics
mental illness
The basic characteristics included age, gender, years of experience, job title, experience in special needs teaching, experience of supporting parents with mental illness, experience of supporting students with mental illness, experience of learning about mental illness, and the history of someone close, who has, or is suspected of having, a mental illness.
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Randomization and blinding
mental illness
BLIND
When school-based applications were received by the research office, a random number table was prepared by a person who was unrelated to the implementation process, and independent allocations were made to either the intervention or control group in the order of arrival of the applications. After the school applications were received, the same research descriptions, which were designed for individuals, were sent to both the intervention and control groups. Because of ethical considerations and the need to ensure that the participants understood the purpose of the study, we simply stated, in the research description, that the children of parents with mental illness might face challenges. After the allocation of the schools, data managers were no longer blind.The next time any schoolteacher’s individual application form arrived at the research office, the schoolteacher was assigned to the group to which the school was assigned. The participants were guided to their allocation group after the application, and they were no longer blinded. The web surveys and the video were sent at pre-set times, using a computer research electronic data capture accumulation and management system. The quantitative data analyses were blinded to the analysis.
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Analyses
As the primary population, the full analysis set (FAS), excluding the scores of those who did not respond to T1 from all randomized participants, was used in all analyses, except process and feasibility evaluation. The per-protocol set (PPS), which excluded participants who did not respond on time, those who did not complete watching video on time in the intervention group, and those who withdrew from the study, was also defined for sensitivity analysis of the primary outcome.Baseline characteristics are summarized as mean and standard deviation for continuous variables and frequency and proportion for categorical variables. Student’s t-test, chi-square test, and Fisher’s exact test were performed for comparisons between the intervention and control groups.The difference in the Sense of Coping Difficulty subscale at T3 between groups was evaluated by a t-test on cluster-specific means. As a sub-analysis, a multilevel analysis with a mixed model was performed, with group as fixed effects, and school as random effects, adjusting for some baseline characteristics. We fitted a mixed model for repeated measures (MMRM), with group, time points (T1, T2, and T3) and their interactions as fixed effects, and schools and study participants as random effects.The three actual behavioral and attitude items were compared between the groups using chi-square analysis. A multilevel analysis was then conducted using a mixed-effects logistic model.For the 10 knowledge items, the percentage of correct answers for each item was compared between the groups using chi-square analysis or Fisher's exact test. The total number of correct answers was compared using a Wilcoxon rank-sum test. Mixed-effect logistic models were also fitted.For program goal achievement, the medians and interquartile ranges of the scores and the total scores for each item were calculated and compared, using the Wilcoxon rank-sum test. MMRM were also fitted.In an exploratory manner, we examined the interaction between the baseline score and intervention effects on the Sense of Coping Difficulty subscale.The open-ended text response was analyzed qualitatively as part of a process evaluation. We created classification axes based on similarity and separated them into the smallest units whose meanings could be understood [The missing data were not imputed. A p-value of 0.05 or less was considered significant. All analyses were performed using SAS9.4 (SAS Institute Inc., Cary, NC, USA).
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Ethical procedures
ADVERSE EVENTS
This study was approved by the Ethics Committee for the Intervention Study of Osaka University Hospital (approval no. 21144; 5/10/2021). Written informed consent was obtained from all the participants. This study was conducted in accordance with the principles of the Declaration of Helsinki. This project has been registered in the Clinical Trial Registry (UMIN000045483; 14/09/2021). No adverse events were reported.While filming one individual in the video, we asked her to decide whether she would like to have her face and name included in the video. She chose to show her face but not disclose her name. We notified the person in writing and obtained her consent for the study participants to watch the video.
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Results
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Study participants
infection
MAY, INFECTION
This study was conducted between October 2021 and May 2022. The flow of study participants is shown in Fig. Flow of study participantsEighty-six participants in the intervention group and 82 in the control group completed T3. Of the 86 participants in the intervention group, 73 became PPS, excluding 8, who responded after the deadline, and 6, who watched after the deadline (with duplicates, a total of 13 participants). Of the 82 participants in the control group, 73 had PPS, excluding nine who responded after the due date.The number of participants did not reach the originally expected number. This result was attributable to the schools’ response to COVID-19. The number of people infected with COVID-19 was declining in Japan in the fall of 2021. However, when this study began, schools reopened after the summer break; although students were able to attend school, schoolteachers were busy due to government orders to strengthen infection control measures in schools [
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Process and feasibility evaluations
In the intervention group, 83 of 86 participants were "very satisfied" or "somewhat satisfied" with the program, and three were "not very satisfied." Regarding whether they would recommend the program to other schoolteachers, 85 were "willing to recommend" and 1 was "unwilling to recommend.” The length of the program was "long" (26 participants), "just right" (60 participants), and "short" (0 participants).In terms of “what you learned was new in the program,” 82 answered “yes” and 4 answered “no.” Among those who agreed, Table Qualitative analysis of open-ended text responseRegarding “whether your way of thinking has changed,” 60 felt that it had, while 26 felt that it had “not changed.” Of those who got “changed,” in order of preference, 19 "want to stay involved with the children," and 16 wrote, "view of the children's background.”Regarding “how you plan to act,” the most common responses were, in descending order, "look carefully at the children's background," written by 22 participants, "continue to stay close to the children," by 20, and, "try to be aware of children in need of support,” written by 19. Related to the school's organizational response, some responded, "entire school responds, not just individuals," "share with the entire school, not just individuals," "create a safe environment at school," and "share with other schoolteachers.”
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Discussion
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Study participants
mental illness, illness
The intervention and control groups had approximately the same number of clusters and schoolteachers, with no significant baseline differences. The mean age of the study participants was 43.6 years (intervention group) and 41.1 years (control group), similar to the mean age of public elementary schoolteachers in Japan, 42.6 years [Additionally, 55.2% (intervention group) and 58.3% (control group) of the study participants were female, slightly less than the 62.4% of all public schoolteachers [The experience of working with parents with mental illness was common to more than half of the participants, but only about 30% had learned about mental illness. Among elementary schoolteachers who resigned because of illness, 69.1% resigned because of mental illness [
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Effectiveness of the program
mental illness
The program was effective in reducing schoolteachers’ difficulty in supporting children of parents with mental illness. The effect sizes were as large as –0.90 (FAS) and –0.86 (PPS). Goal attainment was also effective for over one month. Few studies on mental health interventions for schoolteachers have been conducted using RCTs [Although exploratory in nature, this program was found to be particularly effective for those with higher scores (higher difficulty in support) on the Sense of Coping Difficulty subscale. The program included what children appreciated about their schoolteachers, such as staying close to them and listening to them. In the analysis of the open-ended text response, many schoolteachers planned to continue to stay close to the children. Staying close to children does not require schoolteachers to develop any specialized skills in dealing with mental illness. This may have given schoolteachers the confidence to provide support.Knowledge was effective up to the T3; the correct response rate of “mental illness affects 1 in 5 people in their lifetime (no. 1),” and “half of those affected by mental illness develop the illness by their mid-teens (no. 6).” These items were objective measures of the onset timing and probability of onset. However, no significant effects were found for items not expressed numerically (no. 4, 5, 7, 8, 10). Many previous programs on schoolteachers’ mental health literacy have also been effective in increasing knowledge [For actual behaviors and attitude, only “looked at children from the perspective of a child in need of support” had a significant effect in the past month, whereas “recognized the presence of a child(ren) in need of support” and “consulted within the school about a child(ren) in need of support” had no significant effect. There were significant effects on goal attainment related to confidence at the behavioral level (nos. 4–7). In the analysis of the open-ended text response, “the importance of staying close to children” was the most common impression (21 participants), and “continue to stay close to children” was also the second most common action plan (20 participants). The “view of the children's background” also changed their outlook (16 participants), and they were willing to “look carefully at the children's background” (22 participants). Therefore, it is thought that, although they were able to look at the children from the viewpoint of children in need of support, this was not reflected in changes in their behavior (e.g., recognizing the existence of the children or consulting with the children at school). One possible reason is that the child(ren) in need of support could not be detected in a month. Therefore, a long-term evaluation may help schoolteachers recognize such a child(ren). Another possible reason could be that children are wary of their surroundings because they do not want their parents to be known. A total of 55% of adult children of parents with mental illness reported that they did not show signs that those around them could recognize when they were in elementary school [All program goal achievement scores remained effective up to one month later. We believe that the effect took hold for one month, not only due to the program content but also because of how it was structured. While developing this program, we considered the current lack of social awareness regarding the existence of children with parents having mental illness. Referring to the TTM, the program included self-re-evaluation, and we asked the following questions before providing new knowledge, “Have you ever met a child like this?” Baseline participants’ information indicated that more than half of them had experience working with parents having mental illness. Therefore, it is assumed that the participants recalled children they had met in the past and reflected on their own responses at that time. Furthermore, after watching the independent adult in the video, they may have thought about the aftermath of their own support. It is believed that they could become more confident in their support and more willing to provide support with hope. Though this program involved a short 30-min video, we believe that the structure of this program was also effective in that its effect lasted for one month.
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Program feasibility and adaptation to practice
Although Japanese schoolteachers are very busy, 168 of the 171 baseline participants remained in the study until the end. This may have been due to the short duration of the program (30 min) and the easy participation in the e-learning format, which was not restricted to any location and allowed them to choose their own locations. The length of the program was “long” (26 participants), “just right” (60 participants), and “short” (none). Therefore, although a shorter length would have been more desirable, the current one was considered acceptable. Thirty minutes would be sufficient to set up a training session with additional group work after watching the video. As mentioned in the open-ended text response section, it is sometimes difficult to solve problems unless they are shared and handled by the entire school. The need for an approach that involves the entire school has been pointed out in a previous study [
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Study limitations
This study had several limitations. First, the program assessed the understanding and behaviors of individual schoolteachers but not the organizational response of the school as a whole. There were opinions that in order to identify and support children, not only the efforts of individual schoolteachers, but also the whole school approach is needed. Therefore, in the future, this program should be shared with the entire school and evaluated from an organizational perspective as well. Second, it was assumed that the study participants had some interest in the issue, and those who had no interest in the issue at all would not have participated in this study. Therefore, we do not know whether this program is effective for those who have no interest in the issue. Third, mental health conditions involve a wide spectrum of conditions. This program does not cover all mental illnesses.
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Conclusion
mental illness
We developed a 30-min video-based e-learning program to help elementary schoolteachers support children of parents with mental illness and evaluated its effectiveness using a school-based cluster randomized controlled trial. It was observed that it was significantly effective, over one month, in reducing difficulties in supporting children.
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Acknowledgements
mental illness
We thank all elementary schoolteachers who contributed to our research. I would like to thank the core group members of the self-help group for adult children of parents with mental illness, “KODOMO-PEER.” We also thank Prof. Yoshie Hamashima (Osaka Dental University) and Ms. Natsumi Asada (Taanto Life) for their advice on the program.
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Authors’ contributions
RS
Design of the study: MK, AM; development of the program: MK, AK, TS, YE, AM, SS, HK, SI, KY; development of question items: MK, YE, SS, AM; coordination with research fields: AM, KY, MK; quantitative analysis: RS; qualitative analysis: MK, KK; data management: MK, KK, HK; manuscript preparation: MK. All the authors have checked and confirmed the final manuscript prior to submission. The author(s) read and approved the final manuscript.
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Funding
This study was supported by JSPS KAKENHI (grant numbers JP 19H03960 and 20K10788).
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Availability of data and materials
The materials for this program are openly available (
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Declarations
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Ethics approval and consent to participate
ADVERSE EVENTS
This study was approved by the Ethics Committee for the Intervention Study of Osaka University Hospital (approval no. 21144; 5/10/2021). Written informed consent was obtained from all the participants. This study was conducted in accordance with the principles of the Declaration of Helsinki. This project has been registered in the Clinical Trial Registry (UMIN000045483; 14/09/2021). No adverse events were reported.
PMC10024023