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Funding
This project has been supported by the Research and Technology Deputy of Hamadan University of Medical Sciences (Grant Number: 9904242558).
PMC10422136
References
PMC10422136
Methods
Participants were randomly divided into two groups (n = 35), with the experimental group undergoing art therapy involving weekly 60-min group therapy sessions for 10 weeks. Statistical analysis was performed using Ranked ANCOVA and Wilcoxon’s signed rank test. Western blotting was performed to analyze serum SAP levels.
PMC10162529
Results
depression, impulsivity, anxiety
We observed an association between psychological mechanisms and stress proteins. There was an increased number of NK cells in the experimental group after the program. Moreover, compared with the control group, the experimental group showed significant changes in SAP expression. Further, the experimental group showed a positive change in the MMPI-2 profile, as well as a decrease in depression, anxiety, impulsivity, and alcohol dependence.
PMC10162529
Conclusions
RECURRENCE
Continuous psychological support could be applied as a stress-control program for preventing stress recurrence and post-discharge relapse. Our findings strengthen the link between biomedical science and mental health in rehabilitation treatment for AUD.
PMC10162529
Data Availability
All relevant data are within the paper and its
PMC10162529
Introduction
deaths, cognitive and behavioral disorders
According to the World Health Organization, alcohol is the most consumed psychoactive substance worldwide, with an estimated 3 million deaths annually worldwide due to harmful alcohol use [Chronic alcohol use causes extensive structural, functional, and neurobiological changes in the nervous system, leading to a various problems including addiction, thinking, emotional, cognitive and behavioral disorders [
PMC10162529
AUD, stress proteins, immune cells, and electroencephalography (EEG) changes
Brain injury, sleep disorders, anxiety
RECURRENCE, BRAIN
Prolonged alcohol use significantly decreases leukocyte levels and antibody production. In case of long-term alcohol misuse, alcohol directly acts on tissues involved in stress response, which increases the production of stress-related proteins [In addition to biomarkers, changes in EEG are also important for AUD patients. Brain injury caused by chronic alcohol consumption involves deteriorated brain signaling, physiological and psychological tension through stimulation of the autonomic nervous system, and withdrawal symptoms such as anxiety and sleep disorders [Previous QEEG studies on patients with AUD have primarily focused on the patients’ states rather than post-treatment changes, Moreover, no studies have conducted QEEG before and after an AUD intervention program to observe changes. Brain wave analysis is clinically significant since brain waves are strongly associated with AUD severity and recurrence [
PMC10162529
AUD and the Minnesota Multiphasic Personality Inventory-2 (MMPI-2)
personality traits
In the past decades, personality traits have been identified as important predictors of treatment success and relapse in patients with AUD [
PMC10162529
AUD and AT
depression, brain damage, impulsivity, anxiety
According to the American Art Therapy Association, AT facilitated by a professional art therapist can effectively support personal and relational treatment goals as well as community concerns [Since art can only function following the laws of the visual brain, patients with AUD can be trained to regulate the loss of control and impulsive urges resulting from brain damage [Regarding AT-related brain waves, Belkofer and Konopka reported a marked increase in alpha and beta activities after painting and drawing [We aimed to investigate the effect of AT on emotional (Minnesota Multiphasic Personality Inventory-2 [MMPI-2], alcohol dependence, depression, anxiety, and impulsivity) and physical (natural killer [NK] cell count, expression of stress-associated proteins [SAP], and electroencephalography) changes in patients with AUD.
PMC10162529
Materials and methods
PMC10162529
Participants and procedure
RECURRENCE
The study was conducted from March 2017 to March 2018 at KARF St. Mary’s Hospital in Ilsan, Republic of Korea. We recruited 55 patients with AUD; all met the study criteria and provided informed consent. We randomly allocated the 55 participants into the control (n = 27) and experimental (n = 28) groups. However, 20 participants dropped out due to recurrence of alcohol use. Finally, 15 and 20 participants from the control and experimental groups, respectively, completed the study.
PMC10162529
Randomisation method and allocation concealment
anger
RECURRENCE
This study was a single-blind study; the hospital’s clinical pathologists and doctors in contact with the patients were blinded, while the clinical art therapists conducting the study were not blinded. Participants were randomly assigned to groups using a computerized random number table by university volunteers who did not participate in the study intervention. The research description and informed consent form explained that the probability of being assigned to each group is 50:50. The assignment order was hidden in sequentially numbered, opaque, sealed, and stapled envelopes that did not transmit intense light in which participants were enrolled. Participants’ names and dates of birth were recorded on the envelopes to prevent the dispensing order from being reversed. Results were available after enrolled participants had completed all baseline assessments and before assigning interventions. Both groups received the same hospital-provided addiction treatment (AUD education, cognitive behavioral therapy, anger control training, recurrence prevention training, high-risk situation management program, etc.) (Figs
PMC10162529
Recruitment and randomized allocation group from the CONSORT 2010 flow diagram.
PMC10162529
The overall flow of research.
We observed the changes in NK cell number and SAP expression, emotion, and behavior by applying AT, a complementary alternative medicine (CAM) to the psychological risk factors of AUD. Resulting changes in psychological factors led to positive NK cell growth and changes in SAP expression. It has also been shown that positive changes in the MMPI-2 profile can also change the personality of the individual. This shows the connection between physiology and mental health in addiction rehabilitation and shows the importance of psychological rehabilitation. This study was the first to show the biological basis of the application of complementary therapies, a non-drug treatment method of AUD, to humans and to monitor progress.
PMC10162529
Participants
The inclusion criteria were as follows:Adult patients diagnosed with AUD by a psychiatrist.Capable to participate in AT according to a psychiatristHospitalized in the research facility with completion of the 2-week detoxCapability to perform minimal motor activities required for AT.Having received consent from their guardians to participate in the study.The exclusion criteria were as follows:Pregnant or lactating during the trialVoluntary termination of the study.Difficulty in undergoing further studies due to serious diseasesPatients who did not meet the subject criteria and participated falsely or caused physical and mental damage due to serious self-harm.
PMC10162529
Sample size
This study was a pilot study. Per Sheatsley and Sudman, a pilot study was appropriate with a minimum of 12 to 50 participants prior to a full-scale study [
PMC10162529
Ethical approval
DISEASE
This study was conducted in accordance with the Declaration of Helsinki. Ethical approval was obtained from the CHA University Institutional Review Board in March 2017 (1044308-201612-BR-030-03) (The clinical trial registration number is as follows:Clinical Research Information Service [Internet]; Osong (Chungcheongbuk-do): Korea Centers for Disease Control and Prevention, Ministry of Health and Welfare (Republic of Korea); KCT0004588; December 31, 2019
PMC10162529
Informed consent
All the participants provided written informed consent. The consent form outlined the purpose, process, method, duration, side effects or risks, benefits, and disadvantages of the study. Additionally, the confidentiality of the collected personal information was guaranteed.
PMC10162529
Intervention
EVENTS
A clinical art therapist and two assistant therapists conducted the AT program. The main therapist was an AT expert with over 6 years of experience. The assistant therapists were graduate students in their master’s courses in clinical AT. The assistant therapists explained how to use the materials and helped participants with difficulty expressing their work. Pre- and post-test results, photographs of the work, and consultations were recorded after completion of the program.Only the experimental group underwent AT, which involved weekly 60–90 min group therapy sessions conducted for 10 weeks. Both groups underwent the same post-test 10 weeks after the pretest. AT was administered as group therapy, with each group comprising nine patients. The group therapy was performed in a sufficiently large lecture hall at the study facility. For the first 15 min of the session, the therapist explained the session’s topic; further, the participants talked about recent events about themselves. Subsequently, the participants chose materials and expressed the topic in their art for 40 min. They were allowed to freely communicate during this process; moreover, they concentrated on their work. After completing their artwork, the participants and therapist discussed the drawings of each member for 10–30 min.AT was conducted as part of the intervention program for AUD, which was structured according to the psychological problems of AUD.
PMC10162529
AT intervention program.
PMC10162529
Measures
PMC10162529
Detection of changes in the immune cell count and SAP expression levels
Alzheimer’s disease, depression, Down’s syndrome, drug addiction
BREAST CANCER
A blood test for SAPs allows objective assessment of the positive effect of AT on the immune system and stress levels. We tested for cortisol levels, NK cell count, and SAP expression levels (C-Jun N-terminal kinase [JNK], p-JNK, and Elk-1).Elk-1 is a transcription factor that directly regulates the expression of immediate early genes and is crucially involved in long-term memory, drug addiction, Alzheimer’s disease, Down’s syndrome, breast cancer, and depression [
PMC10162529
Electroencephalography (EEG)
bipolar
EEG was performed using the Neurofeedback System (Panaxtox Corp, Seoul, Korea) developed by the Korea Psychiatry Research Center. This device simultaneously measures the left and right EEG traces in FP1 and FP2 of the prefrontal lobe using two electrodes in a sequential bipolar montage, following the International 10–20 system. A solid electrode was attached to the headband; subsequently, EEG was measured from the left and right frontal lobes through the FP1, FPz, and FP2 channels, which were fixed at 4-cm intervals on the left ear lobe as a round electrode. EEG measurements assess real-time brain function and allow objective brain function analysis. EEG is a frequency-based spectrum analysis method that captures the degree of slow and fast waves through correlation and provides more information than existing analysis methods for each [
PMC10162529
The brain function quotient.
PMC10162529
Attention quotient (ATQ)
diseases or stress [
DISEASE
ATQ represents the degree of brain arousal and resistance to disease or stress. It is calculated by dividing theta wave activity by sensorimotor rhythm (SMR) wave activity of approximately 12–15 Hz. Therefore, it is related to immunity against diseases or stress [
PMC10162529
Activity quotient (ACQ)
The ACQ is an index representing the level of alpha and low-beta wave activity as well as the overall activity in the left and right brain. It is used to determine mental activity, thinking, and behavioral tendencies. It can be computed by analyzing alpha and low-beta waves. High and balanced ACQs between the left and right brains are ideal [
PMC10162529
Minnesota multiphasic personality inventory-2 (MMPI-2)
psychiatric disorders
The MMPI-2 is a widely used self-reporting test developed by Hathaway and McKinely for accurate clinical diagnosis and evaluation of patients with psychiatric disorders [
PMC10162529
Alcoholism screening test of the National Seoul Mental Hospital (NAST)
The NAST was developed by Kim et al. [
PMC10162529
Center of Epidemiologic Studies Depression Scale (CES-D)
depressive
The CES-D was developed by the National Institute of Mental Health in 1971 to measure depressive symptomatology in the general population. We used the Korean version translated by Cho and Kim [
PMC10162529
Beck anxiety inventory (BAI)
Anxiety, anxiety, psychiatric disorders
Anxiety was measured using the BAI, which is a self-report questionnaire developed by Beck, Emery, & Greenberg. It allows for the assessment of clinical anxiety in patients with psychiatric disorders and comprises 21 items regarding cognitive, emotional, and physical anxiety [
PMC10162529
Barratt impulsiveness scale-II (BIS-II)
We used a modified version of the 11
PMC10162529
Data collection and procedures
Both groups underwent baseline assessment before the first session and post-intervention assessment after the final session. The EEG and questionnaire surveys were conducted in a testing room in the study facility without disturbances. Moreover, laboratory tests for the NK cell count, cortisol levels, and protein levels were conducted by a clinical pathologist at the study facility. The EEG was performed within 10 min, including preparation, while the survey, including the MMPI-2, took about 90–120 min. Participants were assigned an identification number, which was used to manage and document their baseline and post-intervention data.
PMC10162529
Statistical analysis
depression, impulsivity, anxiety
Statistical analyses were performed using SPSS Statistics (version 22.0). Specifically, we performed a frequency analysis of the participants’ sociodemographic characteristics. Although normality was assumed through the normality test, a nonparametric test was performed because the number of study participants was small, and an ordinal scale was used. Ranked ANCOVA was performed for the NK cell count and cortisol levels. The MMPI-2, SAP, EEG, depression, anxiety, and impulsivity data were analyzed using Wilcoxon’s signed rank test. Statistical significance was set at
PMC10162529
Western blotting
SAP expression was measured using serum samples obtained from both groups. Bradford assay was performed to measure and quantify protein levels in each sample, followed by protein analysis using Western blotting with anti-JNK (Cat# SC-571 Santa Cruz, Santa Cruz, CA, USA), anti-p-JNK (Cat# SC-571 Santa Cruz, Santa Cruz, CA, USA), and anti-Elk-1 antibodies (Cat# SC-355 Santa Cruz, Santa Cruz, CA, USA). First, the total protein concentration in each serum sample was quantified using the Bradford method. Accordingly, 20 μg of protein was analyzed through sodium dodecyl sulfate-polyacrylamide gel electrophoresis at 120 volts for 1.5 h.Subsequently, proteins in the gel were transferred to a nitrocellulose membrane (Bio-Rad, CA, USA) at 120 Volts for 1 h. The membranes were blocked for 30 min at room temperature with 5% blocking buffer, which was prepared by dissolving 2.5 mg of nonfat milk in 50 ml of washing buffer comprising 10 mM (5 ml of 2M stock) Tris-HCl (pH 7.5), 100 mM (20 ml of 5M stock) NaCl, 1 ml of 0.1% Tween 20, and 976 ml distilled water. The membranes were probed with primary antibodies against SAP, p-SAP, and Elk-1 (all 1:1,000 dilution), followed by incubation at 4°C overnight. After washing with washing buffer, the membranes were incubated for 1 h with horseradish peroxidase-conjugated polyclonal goat anti-rabbit IgG (1:10,000 dilution). The membranes were re-washed with washing buffer and developed on an X-ray film with ECL reagents (enhanced chemiluminescence kit, Youngin Frontier, Seoul, Korea).
PMC10162529
Results
AUD
RECURRENCE
We randomly allocated the 55 participants into the control (n = 27) and experimental (n = 28) groups. However, 20 participants dropped out due to recurrence of alcohol use. Finally, 15 and 20 participants from the control and experimental groups, respectively, completed the study. With the exception of dropouts, all participants proceeded as they were in their initially assigned group. The average age of the samples analyzed for the control and the experimental groups was 46.9 years (SD = 11.7161) and 40.7 years (SD = 8.8858), respectively. The alcohol dependence screening test revealed that most participants were at an advanced AUD stage, with 34.3% and 85.8% scoring above 30.1 and 11, respectively (
PMC10162529
General characteristics of the control and experimental groups.
PMC10162529
Group comparisons
PMC10162529
Changes in the NK cell count, cortisol levels
To determine the effectiveness of AT, Ranked ANCOVA was performed to compare experimental and the control groups in the change scores of NK cell, and Cortisol. The results of Ranked ANCOVA is presented in
PMC10162529
Ranked analysis of covariance for NK cells, and cortisol.
**
PMC10162529
Changes in stress-associated proteins
We analyzed changes in SAP expression in both groups. Repeated western blotting experiments were performed on two randomly selected participants from each group (Figs
PMC10162529
Means and Wilcoxon’s signed-rank test comparisons for study variables.
Depression, anxiety
*** ** * Effect size cut offs: 0.20 small, 0.50 medium, ≥ 0.80 large.ATQ, attention quotient; ACQ, Activity quotient.; Stress proteins (JNK, p-JNK, ELK1); NAST, Alcoholism screening test of the National Seoul Mental Hospital; CED-D, Center of Epidemiologic Studies Depression Scale; BAI, Beck anxiety inventory; BIS- Ⅱ, Barratt impulsiveness scale-II.
PMC10162529
Changes in the EEG, NAST, impulsivity, depression, and anxiety
As shown in In addition, the experimental group decreased alcohol dependence (Z = −3.920,
PMC10162529
Changes in the MMPI-2 profile
The experimental group showed significant changes in the validity scale of the MMPI-2 (F, F (B), L, K, and S) (
PMC10162529
Means and Wilcoxon’s signed-rank comparisons for MMPI-2 (validity scales and clinical scales).
RCd, antisocial tendencies, Hypochondriasis, low impulsive, Psychasthenia, schizophrenia, depression, Depression, obsessive-compulsive disorder
RECURRENCE, PATHOLOGY
** * Effect size cut offs: 0.20 small, 0.50 medium, ≥ 0.80 large.VRIN (Variable Response Inconsistency), TRIN (True Response Inconsistency), F (Infrequency), F(B) (Back Infrequency), F(P) (Infrequency Psychopathology), FBS (Symptom Validity), L (Lie), K (Correction), S (Superlative Self-Presentation), Hs (Hypochondriasis), D (Depression), Hy (Hysteria), Pd (Psychopathic Deviate), Mf (Masculinity-Femininity), Pa (Paranoia), Pt (Psychasthenia), Sc (Schizophrenia), Ma (Hypomania), Si (Social Introversion).Moreover, the experimental group showed significant changes in the clinical scale (D, Pd, Pa, Pt, Sc, and Si), indicating a positive change in the levels of depression, antisocial tendencies, paranoia, obsessive-compulsive disorder, schizophrenia, and introversion.There was a significant change in the levels of demoralizing negative emotions, cynical attitudes, dysfunctional negative emotions, and aberrant experiences in the experimental, but not control, group (RCd, RC2, RC3, RC7, and RC8) (In the personality pathology 5 factor scale, the experimental group showed a significant change in negative emotionality and introversion AT (Twenty patients dropped out of this study, which indicates the difficulty of rehabilitation in addiction treatment and the high recurrence rate. There was a significant change in the DISC score in the control group since it had lower pre-test scores than the experimental group. Since a considerable number of patients in the control group dropped out, those who did not could have been able to cope with the possibility of controlling themselves and had a low impulsive tendency.
PMC10162529
Discussion
drug addiction, anxiety, sleep disturbances, antisocial, aggression, impulsivity, psychasthenia, depression, schizophrenia, irritability, alcohol dependence
This study examined the physical, emotional, and behavioral changes experienced by patients with AUD after AT. First, we found that AT was associated with an increased NK cell count. Alcohol use decreases immune function; moreover, an increased NK cell count indicates increased immune function. The mechanisms through which socio-psychological factors affect immune function and the resulting physiological changes remain unclear [Second, the experimental group showed decreased JNK and p-JNK expression and variations in Elk-1 expression. These findings demonstrate that AT may alleviate stress and depression in patients with AUD. There was a significant between-group difference in SAP expression, which indicates an effect of AT on the body’s anti-inflammatory functions. Elk-1 is associated with drug addiction and depression, with activation by JNK. Future studies should investigate the involvement of Elk-1 in pathophysiological depression [Third, there was a non-significant decrease and increase in cortisol levels in the experimental and control groups, respectively. Cortisol is crucially involved in regulating the limbic and immune systems; moreover, it is associated with changes in behavior, cognition, and immune responses [Fourth, AT caused positive changes in the brain waves of patients with AUD. The ATQ is calculated by dividing the theta wave activity by SMR wave activity (approximately 12–15 Hz) [The ACQ is determined by relative intensity, absolute intensity, log comparison, and arithmetic comparison [The human brain has plasticity, where it consistently adapts and restructures based on learning and experiences [Fifth, the experimental group showed a reduction in depression, anxiety, aggression, impulsivity, alcohol dependence, and anti-sociality. The D scale, which showed the second-highest score in the MMPI-2 in our study, describes the likelihood of suicide attempts, sleep disturbances, vitality, and attention. These effects were markedly alleviated in the experimental group. Further, there were positive changes in DEP, RC7, and RC8, which can be attributed to decreased depression, anxiety, dysfunctional negative emotions, irritability, and aberrant experiences, as well as improved interpersonal relationships. The experimental group showed increased positive emotions and a decreased tendency to feel less positive emotions. There was a positive change in subjective well-being, which alleviated depression and anxiety. Moreover, the experimental group showed decreased aberrant sensory experiences and cynical attitudes. The relationship between AUD and depression is important because it may be a motivation for initial alcohol use. AT had a positive effect on the scores for psychasthenia and paranoia, indicating a relief effect on inappropriate feelings by controlling stress in patients with AUD who lack stress-coping strategies. Consistent with these findings, there was a significant between-group difference in the CES-D and BAI scores.The experimental group showed significant improvement in the BIS-II score, as well as in the antisocial tendency and RC scales in the MMPI-2. Recent studies have shown that impulsivity increases the addiction frequency [Sixth, the experimental group showed improved social relations and self-concept. The MMPI-2, Si, and INTR scales could be associated with depression and social introversion, which causes difficulties in group settings and social life. There were significant changes in the schizophrenia, RC2, and RC3 scales in the experimental group, which indicated positive emotional involvement in life, decreased cynical attitudes, and social depression. AT could eliminate negative emotions through increased attention and acceptance of one’s emotions and others’ feelings, which causes positive changes in anxiety, inferiority, and depression. Group AT can help participants accept other people’s emotions.
PMC10162529
Limitations and suggestions for future research
This study had several limitations. First, we included a relatively small sample size. Given the high dropout rate of participants (due to relapse), there was a between-group difference in the sample size. This could undermine the generalizability of our findings. Second, we did not consider the influence of environmental factors, including the function of the participant’s family or differences in the surrounding environment. Third, we did not perform age- and gender-based comparisons. Finally, we only recruited participants from one research institute.Future large-scale studies with gender-based comparisons are warranted to confirm the effect of AT. Additionally, there is a need for active prospective studies on the effects of psychological health on physical health, post-discharge abstinence, stress coping style, and networking with organizations. The home support system or other social factors can also influence the outcomes; therefore, an appropriate experimental design is required to control for such variables. A comprehensive outlook and approach to AUD are required from multidisciplinary, with the inclusion of both treatment and rehabilitation.
PMC10162529
Conclusions
depression, impulsivity, anxiety
The results of our study show that through AT, alcohol dependence, depression, anxiety, and impulsivity in patients with AUD decreased, and that AT was effective prevention and recovery. The results do not indicate a reduction in the number of patients with AUD but do suggest that AT is indeed beneficial in numerous ways.AT can be used as a stress control program for preventing relapse and can help patients with AUD become functional members of society. We found that psychological support through AT had a significant effect on immune response, stress response, and psychosocial measures. And by observing the changes, we found a link between psychological mechanisms and SAP. This study is the first of its kind to present biological evidence in the treatment of psychological problems that could have a physically negative impact on AUD patients in humans, and shows the connection between physiology and mental health in addiction rehabilitation and shows the importance of psychological rehabilitation. We hope that the results of this study would contribute to its applicability of AT as one of the intervention methods for treating AUD.
PMC10162529
Supporting information
PMC10162529
CONSORT 2010 checklist of information to include when reporting a randomised trial*.
(DOC)Click here for additional data file.
PMC10162529
Original images for blots and gels.
(PDF)Click here for additional data file.
PMC10162529
Restructured clinical scales and Psy-5 scales T-scores were compared by the group.
(DOCX)Click here for additional data file.
PMC10162529
Content scales of T-scores were compared by the group.
(DOCX)Click here for additional data file.
PMC10162529
Supplementary scales of T-scores were compared by the group.
(DOCX)Click here for additional data file.
PMC10162529
CHA University Institutional Review Board Form- English.
(PDF)Click here for additional data file.
PMC10162529
CHA University Institutional Review Board Form- Korean.
(PDF)Click here for additional data file.We would like to thank the clinical art therapists, Hyon-Suh Kim, Ji-Eun Kim; social worker, Kyung-Ho Jang; and the medical laboratory technologist, Hyang-suk Jeong at the KARF St. Mary’s Hospital who helped us with the research.
PMC10162529
References
PMC10162529
Background
metabolic disease, Diabetes mellitus, diabetic, diabetes
METABOLIC DISEASE, DIABETES MELLITUS, COMPLICATIONS, DIABETES
Diabetes mellitus is a prevalent metabolic disease in the world. Previous studies have shown that anesthetics can affect perioperative blood glucose levels which related to adverse clinical outcomes. Few studies have explored the choice of general anesthetic protocol on perioperative glucose metabolism in diabetes patients. We aimed to compare total intravenous anesthesia (TIVA) with total inhalation anesthesia (TIHA) on blood glucose level and complications in type 2 diabetic patients undergoing general surgery.
PMC10410792
Methods
diabetic
REGRESSION, COMPLICATIONS
In this double-blind controlled trial, 116 type 2 diabetic patients scheduled for general surgery were randomly assigned to either the TIVA group or TIHA group (n = 56 and n = 60, respectively). The blood glucose level at different time points were measured and analyzed by the repeated-measures analysis of variance. The serum insulin and cortisol levels were measured and analyzed with t-test. The incidence of complications was followed up and analyzed with chi-square test or Fisher’s exact test as appropriate. The risk factors for complications were analyzed using the logistic stepwise regression.
PMC10410792
Results
The blood glucose levels were higher in TIHA group than that in TIVA group at the time points of extubation, 1 and 2 h after the operation, 1 and 2 days after the operation, and were significantly higher at 1 day after the operation (10.4 ± 2.8 vs. 8.1 ± 2.1 mmol/L;
PMC10410792
Conclusions
diabetic
COMPLICATIONS
TIVA has less impact on perioperative blood glucose level and a better inhibition of cortisol release in type 2 diabetic patients compared to TIHA. A future large trial may be conducted to find the difference of complications between the two groups.
PMC10410792
Trial registration
The protocol registered on the Chinese Clinical Trials Registry on 20/01/2020 (ChiCTR2000029247).
PMC10410792
Keywords
PMC10410792
Introduction
hyperglycemia, diabetic, diabetes
HYPERGLYCEMIA, DIABETES
It was reported that 8.3% of the population in the United States is diabetic, while in China, the estimated prevalence of diabetes in adults is 10.9% [The effect of anesthetics on the intraoperative blood glucose level and the degree of the surgical stress caused by anesthetic agents plays a crucial role in the postoperative recovery of the patients. Several studies from animal and human have revealed that volatile anesthetics could impair glucose tolerance and suppress insulin secretion, which ultimately resulted in perioperative hyperglycemia [A recent retrospective study showed that sevoflurane and propofol were comparable in terms of incidence and clinical outcomes with respect to perioperative hyperglycemia in diabetes patients undergoing pulmonary surgery [
PMC10410792
Methods
PMC10410792
Trial design
WEST
This randomized controlled single-center, double-blind, parallel trial was approved by the Biomedical Ethics Committee of West China Hospital of Sichuan University on 18/12/2019 (2019 − 928), and written informed consent was obtained from all subjects participating in the trial. The trial was registered prior to patient enrollment on the Chinese Clinical Trials Registry on 20/01/2020 (ChiCTR2000029247). Our study was conducted based on a prespecified protocol from January to October 2020 in accordance with the Helsinki Declaration-2013 [
PMC10410792
Participants
hepatic and/or renal dysfunction, pancreatic cancer, metabolic disorders
DIABETIC KETOACIDOSIS, SYSTEMIC DISEASE, DIABETIC NEUROPATHY, ISLET CELL TUMOR, NEUROMUSCULAR DISEASE, HYPERGLYCEMIA, PANCREATIC CANCER, METABOLIC DISORDERS, TYPE 2 DIABETES
Potentially eligible patients were screened according to the inclusion and exclusion criteria. All adult type 2 diabetes patients (18–90 year old) with class I, II, or III based on the American Society of Anesthesiologists (ASA) physical status undergoing elective general surgery (≥ 2 h) were screened for inclusion according to the study protocol. Exclusion criteria included severe systemic diseases, metabolic disorders, diabetic ketoacidosis or hyperglycemia, diabetic neuropathy, hepatic and/or renal dysfunction, neuromuscular disease, and pancreatic cancer or islet cell tumor. All participants had to understand, express, and write Chinese and give written informed consent.
PMC10410792
Randomisation and blinding
RECRUITMENT, BLIND
The patients were randomized at 1:1 ratio to receive total intravenous anesthesia (TIVA group) or total inhalation anesthesia (TIHA group) by using the SPSS software. The center for Evidence-Based Medicine provided sequentially numbered opaque sealed envelopes to achieve allocation concealment. These sealed envelopes were kept by the screener who was not implementation of the interventions. The envelopes corresponding to the patient’s serial number were sent to the anesthesiologist after recruitment. Then the anesthesiologist opened the envelope and administered the appropriate anesthesia protocol (TIVA or TIHA) according to the card.The randomization allocation schedule was blinded to the outcome evaluators who were not involved in the implementation of the interventions. Participants were blinded to the allocation and did not know the anesthetic protocol they received. It was impossible to blind anesthesiologists from the interventional schemes. However, anesthesiologists were not involved in the measurement or statistical analysis of outcomes and were required not to reveal the details of the anesthetic protocol to outcome evaluators or participants. The statistician was blinded to the group allocation until completion of the statistical analyses. The randomization allocation schedule was also blinded to the outcome evaluators, participants and statisticians until completion of the study.
PMC10410792
Intervention and measurement
hypoglycemia, pain, diabetes
HYPOGLYCEMIA, DIABETES
Oral antidiabetic medications were continued in diabetes patients on the day before surgery, and were discontinued on the day of surgery. The doses of long-acting insulin administered should be reduced by 50–75% on the night before surgery to avoid hypoglycemia during a prolonged fast. Half doses of basal insulin were administered the morning of surgery [The patient was catheterized on radial artery under local anesthesia before induction, and blood was collected to measure the baseline data of relevant outcomes (blood glucose, insulin, and cortisol levels). Midazolam 0.04 mg/kg, sufentanil 0.4 µg/kg, propofol 2 mg/kg and cis-atracurium 0.2 mg/kg were administered for induction of conventional general anesthesia. Endotracheal intubation was performed after mask-assisted ventilation for 3 min.For patients in the TIHA group, anesthesia was maintained with inhaled desflurane (1.0 to 1.5 minimum alveolar concentrations) and intravenous remifentanil infusion (0.12 to 0.24 µg·kgIntraoperatively monitored parameters included heart rate, invasive blood pressure, pulse oxygen saturation (SPatients controlled analgesia, with a sufentanil infusion pump was used in the first 72 h after surgery. Dezocine were used for pain relief 3 days after the surgery. Postoperative use of hormones, non-steroidal anti-inflammatory drugs were avoided.If the intraoperative glucose level was much higher than 10 mmol/L, an advisable dose of insulin should be given without causing hypoglycemia [
PMC10410792
Outcomes
death, postoperative pulmonary complications, arrhythmia, PPCS, stroke, infection, anastomotic fistula, acute kidney injury
MYOCARDIAL INFARCTION (MI), ARRHYTHMIA, POSTOPERATIVE COMPLICATIONS, STROKE, RENAL FAILURE, POSTOPERATIVE COMPLICATION, HYPOGLYCEMIA, STRESS ULCER, INFECTION, HYPERGLYCEMIA, COMPLICATIONS
The primary outcomes are the blood glucose levels at different time points: preoperative(T0), immediate intubation (T1), skin incision (T2), 1, 2 and 3 h after the start of the operation (T3, T4 and T5, respectively), suturing (T6), extubation (T7), 1 h after the operation (T8) ,2 h after the operation (T9), 1 and 2 days after the operation (T10, T11). ​Secondary outcome measures included serum insulin and cortisol levels 30 min before and 30 min after surgery, and the incidence of postoperative complications. The complications were assessed by bedside or over telephone on the 1st, 3rd, 7th, and 30th post-operative day, including myocardial infarction (MI), stroke, acute kidney injury, renal failure, anastomotic fistula, stress ulcer, incision infection, arrhythmia, hypoglycemia, hyperglycemia, postoperative pulmonary complications (PPCS), and death. These postoperative complications were related to the functions of vital organs such as heart, lung, brain and kidney, and were positively associated with intense stress response during surgery. Postoperative complications were assessed according to the criteria from the European Perioperative Clinical Outcome (EPCO) and PPCS definitions [
PMC10410792
Sample size
The sample size was corrected for repeated measures by blood glucose level at 11 different time points based on the results of the pilot trial with SPSS software. According to the pilot trial, the blood glucose level on the postoperative day 1 was largely adequate to prove a difference between the two groups, and the mean [SD] blood glucose levels were (TIVA 10.9 ± 2.2 vs. TIHA 11.7 ± 2.4 mmol/L ). The method used for sample size determination was matched-pair design formula with a type I error of 5% (α = 0.05) and 90% power (β = 0.1). The result showed that at least 51 patients were required in each group to show the differences of blood glucose level between the groups. Considering a 10% loss-to-follow-up rate, the final sample size was determined to be minimally 56 patients each group.
PMC10410792
Statistical analysis
The Statistical analysis of our study was achieved based on a prespecified analysis plan [
PMC10410792
Results
RECRUITMENT
There were164 patients from January 2020 to October 2020 were screened and 134 participants were enrolled in our trial. A Consolidated Standards of Reporting flow diagram of study recruitment was shown in Fig.  Flow CONSORT diagram of study recruitment TIHA: total intravenous anesthesia, TIHA: total inhalation anesthesia
PMC10410792
Patient characteristics
COPD
CHRONIC OBSTRUCTIVE PULMONARY DISEASE, COPD
The baseline characteristics of the patients are summarized in Table  Basic CharacteristicsThe data are expressed as n (%) or median [range]. TIVA, total intravenous anesthesia; TIHA, total inhalation anesthesia; ASA, American Society of Anesthesiologists; COPD, chronic obstructive pulmonary disease; HBA1c, hemoglobinA1c Perioperative VariablesThe data are expressed as n (%) or median [range]. The
PMC10410792
Blood glucose level
BLOOD
The results showed no interaction between time point and group allocation ( Blood glucose level at different time point in the two groups TIHA: total intravenous anesthesia, TIHA: total inhalation anesthesia,
PMC10410792
Insulin and cortisol level
Preoperative insulin level in both groups were comparable (11.1 ± 3.9 vs. 11.3 ± 3.4 IU/mL; Insulin and cortisol levels of the patients in the two groups TIHA: total intravenous anesthesia, TIHA: total inhalation anesthesia,
PMC10410792
Complications and risk factors
PPCS, Postoperative pulmonary complications
REGRESSION, COMPLICATIONS, POSTOPERATIVE COMPLICATIONS
With regards to the postoperative complications, no significant difference was found between the two groups in terms of incidence number of the postoperative complications (TIVA 16% [9/56] vs. TIHA 28% [17/60]; Table  Incidence of postoperative complications in Patients Who Received Different Types of General AnesthesiaThe data are expressed as n (%). The P values were calculated by the Fisher exact test or chi-square test. TIVA, total intravenous anesthesia; TIHA, total inhalation anesthesia; PPCS, Postoperative pulmonary complications; There were no significant differences in overall complications in patients received Different Types of General Anesthesia Binary logistic regression analysis for the postoperative complications (30d)Group
PMC10410792
Discussion
diabetes mellitus, diabetic, lower blood glucose, type 2 diabetes, trauma
POSTOPERATIVE COMPLICATIONS, DIABETES MELLITUS, INSULIN RESISTANCE, TYPE 2 DIABETES, PATHOGENESIS, COMPLICATIONS
In our study, patients with type 2 diabetes in propofol group had lower blood glucose level, less-affected insulin resistance, and better inhibition of cortisol release compared to those in desflurane group. However, no significant difference regarding the incidence of complications between the two groups was found based on the current samples. The pathogenesis of diabetes mellitus is a complicated process, and other systems of the body such as circulatory system, respiratory system and nervous system may be affected at the later stage [Majority of earlier studies focused on the effects of propofol and volatile anesthetics on perioperative blood glucose in non-diabetic patients have concluded that the effect of propofol on intraoperative glucose metabolism was less than that of isoflurane and sevoflurane [As an influenced factor of surgical stress response, blood glucose indirectly reflects the intensity of stress response [Cortisol is one of the stress hormones that reflects the intensity of the body’s stress response. Cortisol secretion sensitively varies based on the severity and duration of the surgical trauma throughout the body [Previous studies suggested that patients with propofol maintenance anesthesia had a longer postoperative survival time than those with desflurane maintenance anesthesia [A limitation of our study that need to be mentioned is the sample size. The sample size in our study was calculated for blood glucose level comparison, which may not be large enough to show the difference of complications, especially long-term postoperative complications between the two groups. A larger sized trial may be needed to find the difference of postoperative complications between the two group in type 2 diabetic patients.
PMC10410792
Conclusions
diabetic
COMPLICATIONS
TIVA has less impact on perioperative blood glucose level and a better inhibition of cortisol release in type 2 diabetic patients undergoing general surgery compared to TIHA. Due to the relative-short term of follow up, it is impossible to compare postoperative survival time in our study. A future large trial may be conducted to find the difference of long-term complications in type 2 diabetic patients between TIVA and TIHA.
PMC10410792
Acknowledgements
Not applicable.
PMC10410792
Authors’ contributions
PL, CZ and CC contributed to the conception and design of the work; XHX, YH and QZ contributed to statistical analysis; XHX contributed to drafting the manuscript; PL and CC revised it. All authors read and approved the final manuscript.
PMC10410792
Funding
This work was supported by the grants No. 81600918 (to P.L.) and No. 81974164 (to C.Z.) from National Natural Science Foundation of China (Beijing, China).
PMC10410792
Data Availability
The datasets used and/or analyzed during the current study are available from the corresponding author on reasonable request.
PMC10410792
Declarations
PMC10410792
Ethics approval and consent to participate
WEST
The protocol for this trial was approved by the Biomedical Ethics Committee of West China Hospital of Sichuan University on December 18, 2019 (2019 − 928), and written informed consent was obtained from all subjects participating in the trial. The trial was registered prior to patient enrollment on the Chinese Clinical Trials Registry on January 20, 2020 (ChiCTR2000029247).
PMC10410792
Consent for publication
Not applicable.
PMC10410792
Competing interests
The authors declare that they have no competing interests.
PMC10410792
References
PMC10410792
Objective
thyroid carcinoma
THYROID CARCINOMA, THYROID
Edited by: Kyung Tae, Hanyang University, Republic of KoreaReviewed by: Mehmet Haciyanli, Izmir Katip Celebi University, Türkiye; Jiajie Xu, Zhejiang Provincial People’s Hospital, China†These authors have contributed equally to this workThis article was submitted to Thyroid Endocrinology, a section of the journal Frontiers in EndocrinologyTo evaluate whether no drainage has an advantage over routine drainage in patients with thyroid carcinoma after unilateral thyroid lobectomy and central neck dissection.
PMC10154604
Methods
thyroid cancer
CAVITY, POSTOPERATIVE COMPLICATIONS, SCAR, THYROID CANCER
A total of 104 patients with thyroid cancer who underwent unilateral thyroid lobectomy and central lymph node dissection were randomly assigned into no drainage tube (n=52) and routine drainage tube (n=52) placement groups. General information of each patient was recorded, including the postoperative drainage volume/residual cavity fluid volume, postoperative complications, incision area comfort, and other data, and the thyroid cancer-specific quality of life questionnaire (THYCA-QoL) and patient and observer scar assessment scale (POSAS) were evaluated after surgery. At the 3–6 month follow-up exam, the differences between the two groups were compared based on univariate analysis.
PMC10154604
Results
Hashimoto’s thyroiditis, numbness, pain, infection, effusion
POSTOPERATIVE COMPLICATIONS, HYPOPARATHYROIDISM, POSTOPERATIVE BLEEDING, INFECTION, EFFUSION, LYMPHATIC LEAKAGE, SCARRING
Significant differences were not observed in the general and pathological information (including sex, age, body weight, body mass index (BMI), incision length, specimen volume, Hashimoto’s thyroiditis, and number of lymph nodes dissected), operation time, and postoperative complications (postoperative bleeding, incision infection, lymphatic leakage, and temporary hypoparathyroidism) between the two groups. The patients in the non-drainage group had a shorter hospital stay (2.11 ± 0.33 d) than the patients in the drainage group (3.38 ± 0.90 d) (P<0.001). The amount of cervical effusion in patients in the non-drainage group (postoperative 24h: 2.20 ± 1.24 ml/48 h: 1.53 ± 1.07 ml) was significantly less than that in the drainage group (postoperative 24 hours: 22.58 ± 5.81 ml/48 h: 36.15 ± 7.61 ml) (all P<0.001). The proportion of incision exudation and incision numbness in the non-drainage group was lower than that in the drainage group (all P<0.05), and the pain score (VAS) and neck foreign body sensation score (FBST) decreased significantly (P<0.05). During the 3- and 6-month follow-up exams, significant differences were not observed between the THYCA-QoL and drainage groups and the non-drainage group, although the scarring and POSAS values were lower than those in the drainage group. In addition, the length of stay and cost of hospitalization in the non-drainage group were lower than those in the drainage group (P<0.05).
PMC10154604
Conclusion
Routine drainage tube insertion is not needed in patients with unilateral thyroid lobectomy and central neck dissection.
PMC10154604
Introduction
thyroid carcinoma, infections, necrotic
THYROID CARCINOMA, SCAR, POSTOPERATIVE COMPLICATIONS, INFECTIONS, NECROTIC
Surgical drainage is a technique used to remove exudate, necrotic tissue, or other abnormally increased fluids from the body through drainage tubes and strips, and it is usually used in the clinical surgical treatment of wounds or after surgery to prevent incision infections and promote wound healing (Therefore, our study compared the differences in drainage/effusion volume, postoperative complications, length of stay, cost, thyroid cancer-specific quality of life questionnaire (THYCA-QoL) and patient and observer scar assessment scale (POSAS) between drainage and no-drainage groups and further evaluated the necessity of routine drainage for patients with thyroid carcinoma after unilateral thyroid lobectomy and central neck dissection.
PMC10154604
Materials and methods
dyspnea, throat, numbness, pain, infection, effusion, hypertension, diabetes
POSTOPERATIVE COMPLICATIONS, HYPOPARATHYROIDISM, CHRONIC DISEASES, POSTOPERATIVE BLEEDING, HYPERTHYROIDISM, MAY, INFECTION, EFFUSION, LYMPHATIC LEAKAGE, LYMPHOCYTIC THYROIDITIS, HYPERTENSION, DIABETES
Patients who underwent unilateral lobectomy and neck lymph node dissection at the First Hospital of Jilin University from November 2021 to May 2022 were selected for this study. Patients were randomly divided into two groups by opaque envelope method: a non-drainage tube after operation group and a routinely placed drainage tube after operation group (The type of drainage in the drainage group was closed negative pressure drainage, which is placed after thyroidectomy, and punctured out 1cm away from the incision).Patients with the following conditions were excluded: those who did not agree to undergo the study, those with prior thyroid surgery, those with a history of hyperthyroidism, and those with systemic chronic diseases, such as hypertension and diabetes. A total of 104 patients were enrolled in this study (n=104) (Trial profile: CONSORT analysis.The general data of the patients were recorded, including the sex, age, body weight, body mass index (BMI), incision length, hospitalization days, hospitalization costs, and operation duration. For patients with drainage after thyroidectomy, when the drainage volume is less than 10ml within 24 hours, the drainage tube is removed, and the patients discharged. The pathological information of the patients was recorded, including the specimen volume, number of lymph nodes dissected, and presence of lymphocytic thyroiditis. The amount of postoperative drainage or cervical effusion and postoperative complications, such as postoperative bleeding, incision infection, lymphatic leakage, sound changes, and hypoparathyroidism, were recorded. For patients without drainage after thyroidectomy, their neck effusion was estimated by ultrasound by measuring the three-dimensional maximum diameter of the hypoechoic area of the thyroid bed in the neck. The patient with symptoms such as neck swelling and dyspnea within 24 hours after surgery which need re-operation was defined as postoperative bleeding. Patients with parathyroid hormone levels below 15.0pg/ml on the first day after surgery was defined as temporary hypoparathyroidism. The overall comfort of the incision area was determined using the pain score (VAS), incision exudation (Some patients were found to have exudation on the surgical dressing during the routine observer after surgery), foreign body sensation in the throat score (FBST), and incision numbness. The quality of life and cosmetic satisfaction of the patients were evaluated using the THYCA-QoL and POSAS values at 3 months and 6 months, respectively.
PMC10154604
Statistical analysis
SPSS version 23 software (SPSS Inc., Chicago, IL, USA) was used for all the statistical analyses. The patient counting data were tested by Pearson chi-square test, the metrological data were tested by the normality test, the data that conformed to a normal distribution were tested by the independent sample’s t-test, and the remaining data were tested by the Mann-Whitney U test.
PMC10154604
Results
throat, foreign-body sensation, Non-drain, effusion, thyroid cancer
SCAR, POSTOPERATIVE COMPLICATION, EFFUSION, THYROID CANCER
The general features and pathological information of patients in the non-drainage and drainage groups were compared (Comparison of general information and pathological information between the non-drainage and drainage groups.The amount of cervical effusion in patients in the non-drainage group was calculated using postoperative cervical ultrasound (Measurement of cervical effusion by ultrasound.Comparison of postoperative complication between the non-drainage group and drainage group.Bold P-Values: p<0.05.As shown in Comparison of hospitalization information between the non-drainage and drainage groups.Bold P-Values: p<0.05.As shown in Comparison of Comfort around neck incision between the non-drainage group and drainage group.VAS, Visual Analog scale; FBST, The foreign-body sensation in the throat score.Bold p-Values: p<0.05.We followed up by presenting THYCA-QoL and PASAS questionnaires to 104 patients at 3-6 months after surgery, among which seven patients were lost to follow-up. As shown in Comparison of THYCA-QOL between the non-drainage group and drainage group.THYCA-QOL, thyroid cancer specific quality of life of questionnaire.Bold P-Values: p<0.05.Comparison of THYCA-QOL between Non-drain group and Drain group. P= 0.781, 0.394.As shown in Comparison of POSAS between the non-drainage group and drainage group.POSAS, patient and observer scar assessment scale; PSAS, patient scar assessment scale; OSAS, observer scar assessment scale.Bold P-Values: p<0.05.Comparison of POSAS between Non-drain group and Drain group. Both P<0.05.
PMC10154604
Discussion
Hashimoto’s thyroiditis, acute postoperative bleeding, postoperative hematoma, numbness, pain, postoperative infection
SCAR, POSTOPERATIVE COMPLICATIONS, BLOOD CLOTS, WOUND INFECTION, RETROSTERNAL GOITER, POSTOPERATIVE BLEEDING, POSTOPERATIVE INFECTION, CAVITY, EFFUSION, SCARRING, LYMPHOCYTIC THYROIDITIS, COMPLICATIONS
Drainage is widely used by many surgeons during thyroid surgery. Many surgeons use drainage tube for every patient after thyroid surgery under the belief that the use of drainage devices can help eliminate exudation and dead space and identify earlier the occurrence of acute postoperative bleeding, which would allow for timely intervention. However, most American and European medical centers do not routinely place drainage tubes for thyroid surgery.Reported have indicated that routine placement of drainage devices after thyroid surgery cannot improve the prognosis of patients but rather increases the risk of postoperative infection and discomfort in the incision area (In this study, significant differences were not observed between the two groups in terms of sex, age, weight, BMI, incision length, specimen volume, number of cleared lymph nodes, and proportion of patients with lymphocytic thyroiditis. The similarity reduces the confounding factors that affect other variables. In addition, the surgeon did not know until after the operation, which ensured that bias was reduced. Because of the surgeons were blinded to the assignment until the end of the operation, surgical conduct would not have been influenced by the assignment.Compared with the drainage volume of the drainage group patients, the cervical effusion volume was significantly reduced in the non-drainage group. This indicates that the existence of a drainage tube may lead to an increase in postoperative exudation, which may be related to stimulation by the drainage tube. The drainage tube stimulates serosa exudation, resulting in a significant increase in drainage in patients in the drainage group (For patients exhibiting acute postoperative bleeding, the drainage device frequently does not draw blood in time due to blockage by blood clots (Studies have shown that routine placement of a drainage tube after thyroid surgery not only increases the risk of postoperative hematoma, wound infection, and other complications (Compared with the non-drainage group, neck discomfort was higher in the drainage group during hospitalization because implanting a drainage device is an invasive operation from the thyroid residual cavity to the outside body, which increases the pain and numbness of the surgical incision in the drainage group patients and a sensation of a foreign body in the neck. The exudation of the residual cavity of the patient follows the drainage tube to the body, resulting in a wetness of the dressing, and the drainage group patients indicated an increase in discomfort. Due to the significant reduction in exudation in patients in the non-drainage group and their faster recovery and increased neck comfort, the hospitalization days and costs were decreased accordingly.In thyroid surgery, the aesthetic needs of patients have become one of the most important factors affecting patient satisfaction because of the specific location of the incision. The placement of the drainage tube at the incision site has a negative effect on incision healing. If the drainage tube is drawn from outside the incision, it will leave an additional surgical scar at site of the incision. This not only reduces patient comfort but also affects the overall aesthetics of the patient’s neck (Not placing a drainage tube after thyroid surgery may increase certain complications. For example, in patients with larger wound surgery, such as retrosternal goiter and Hashimoto’s thyroiditis, it may be necessary to place a drainage tube for better postoperative observations. However, this view is controversial, and some studies have suggested that postoperative thyroid drainage is not associated with the extent of the surgical wound (In conclusion, not placing a drainage tube after surgery improved outcomes for patients after unilateral thyroid lobectomy and central neck dissection and could benefit patients because of the shorter hospitalization times, lower hospitalization costs, less postoperative discomfort, and fewer scarring issues without increasing the incidence of postoperative complications.
PMC10154604
Data availability statement
The original contributions presented in the study are included in the article/Supplementary Material. Further inquiries can be directed to the corresponding author.
PMC10154604
Ethics statement
The studies involving human participants were reviewed and approved by Medical Ethics Committee of the First Hospital of Jilin University. The patients/participants provided their written informed consent to participate in this study.
PMC10154604
Author contributions
Study concept: QZ, ZW. Study design: ZW. Project management: QZ, ZW, PQ. Data collection: ZW, LZ, ZB, XL, QS. Data statistics and analysis: ZW, PQ, LZ. Manuscript preparation: ZW. Manuscript editing: ZW, PQ. Manuscript review: QZ, ZW, PQ. All authors contributed to the article and approved the submitted version.
PMC10154604