This study is designed as a randomized controlled trial (RCT) using block randomization. We will recruit 90 participants diagnosed with aMCI and allocate them into 3 groups in a 1:1:1 ratio; specifically, the Gold Needle “Regulating Spirit” group, the general acupuncture “Regulating Spirit” group, and a sham acupuncture control group. Additionally, we will include 20 healthy volunteers to serve as a control group. The participants’ gender, age, and educational background will be aligned with those of the patients with aMCI. A total of 90 patients with aMCI will undergo treatment for 12 weeks in groups, receiving interventions 3 times per week. Data collection for all enrolled individuals will occur at baseline (week 0), week 12, and week 24.

The study protocol has been registered in ClinicalTrials [18]. The flowchart of the trial is shown in Figure 1 and the research content at different periods of the trial is given in Table 1.

This study used a prospective, RCT design. Individuals with memory impairment were recruited through hospital referrals, poster advertisements, and community screenings conducted by collaborating organizations. Initial screenings identified individuals who met the inclusion criteria for the study’s aMCI cohort. All participants provided voluntary written informed consent prior to the commencement of the study.

This study used a block-randomized design, wherein an independent statistician—who was not involved in trial assessment, treatment allocation, or statistical analysis—generated the randomization sequence using SAS 9.2 (SAS Institute) software’s PROC PLAN (with a block length of 6). An independent research assistant ensured allocation concealment through the use of opaque, sealed envelopes. In this study, the acupuncturist, outcome evaluator, and data analyst were assigned by different individuals to ensure objectivity; both patients and evaluators remained blinded throughout the process. Given the unique nature of Gold Needle manipulation, challenges arose in establishing acupoint settings for the sham group and in maintaining blinding for the acupuncturist. To address these issues, we strictly adhered to patient blinding protocols and scheduled individual appointments at staggered times.

The variables examined in this study encompass both demographic and clinical characteristics, as well as the primary and secondary outcome measures. The fundamental characteristics of each group were assessed at baseline (wk 0). The primary and secondary outcome measures were evaluated at baseline, at the conclusion of the intervention (wk 12), and again at 24 weeks post follow-up.

Recruiters will use a self-constructed questionnaire to collect demographic information from participants, including gender, age, educational attainment, marital status, living arrangements, occupation, and socioeconomic status, along with their medical history and substance use.

The Montreal Cognitive Assessment (MoCA) was used to assess the patients’ overall cognitive function [19]. MoCA encompasses several cognitive domains, including Visuospatial or Executive Function, Attention and Calculation, Naming, Delayed Recall, Orientation, Language, and Abstract Reasoning. The total score ranges from 0 to 30 points, with higher scores indicating superior functionality.

Neuropsychiatric symptoms and a decline in functional abilities are significant concomitant manifestations of AD, with their emergence serving as an indicator that aMCI is progressing toward AD [20]. Consequently, this study incorporated the Functional Activities Questionnaire, along with the Hamilton Anxiety Scale and the Hamilton Depression Scale [21,22].

Acquire multimodal magnetic resonance imaging (MRI) data, including 3D-MRI, resting-state functional MRI, and diffusion tensor imaging, to construct networks related to cognitive function and brain structure [23]. The study participants, who underwent imaging, were scanned using a Siemens Novus 3.0T MRI system, which was manufactured in Germany.

Using single-molecule immunoarray technology for the detection of biomarkers in plasma, we assess the intervention level of the gold standard concerning the Aβ42/Aβ40 ratio in peripheral plasma, alongside measurements of plasma neurofilament light chain, plasma P-tau181, and plasma P-tau217 [24,25].

Throughout the study, the acupuncturist systematically monitored, documented, and communicated adverse events (AEs) using open-ended inquiries. Within 24 hours of the AE occurrence, it was categorized as either treatment-related—common examples being subcutaneous hematoma, persistent pain postneedling, and syncope—or treatment-unrelated. Follow-up continued until the AE was adequately resolved.

This study represents the first application of Gold Needle intervention for the treatment of aMCI. Due to the absence of comparable studies in this domain previously, an accurate estimation of the sample size parameters was not feasible. This study seeks to undertake an exploratory RCT. This study took a group of patients with MCI as the research subjects. In accordance with the sample size recommendations for preclinical trials in traditional Chinese medicine and taking into account parameter precision, a total of 90 participants were selected, comprising 30 individuals in the Gold Needle “Regulating Spirit” group, 30 in the general acupuncture “Regulating Spirit” group, and 30 in the sham acupuncture control group.

Quantitative data were statistically described using SPSS 21.0 software (IBM Corp) and presented as mean (SD; x±s). Three sets of continuous variable statistics were analyzed: Based on the distribution characteristics of the data, comparisons among groups of continuous variables were conducted using either ANOVA or the Kruskal-Wallis H test. In cases where a statistically significant overall difference was identified among the 3 groups, post hoc multiple comparisons were performed using either the least significant difference test or the Nemenyi test. Baseline data, along with data collected at week 12 and follow-up data at 24 weeks, were analyzed using repeated measures ANOVA or generalized linear mixed models tailored for repeated measures.

The brain structure images acquired through MRI will be processed using MATLAB (MathWorks) [26], CAT12 (Computational Anatomy Toolbox; C. Gaser, Structural Brain Mapping Group, University of Jena, Germany) [27], and FSL (FMRI Software Library 5.0; Analysis Group, FMRIB, University of Oxford, United Kingdom). We used 2 distinct analytical approaches, namely voxel-based analysis and tract-based spatial statistics, to assess alterations in total gray and white matter volumes as well as white matter fiber tracts in a patient with aMCI.

Resting-state functional MRI data will be processed using version 2.3 of the WFU_PickAtlas, developed by the Functional MRI Laboratory at Wake Forest University School of Medicine, which is designed to extract the hippocampal region of interest. Use Conn [28] for data preprocessing and statistical analysis. Use the Conn (McGovern Institute for Brain Research) and SPM12 software (Wellcome Centre for Human Neuroimaging) to perform statistical analysis of the results, including functional connectivity, regional homogeneity, and amplitude of low-frequency fluctuations indicators, and visualize the nodes in 3D using the BrainNet Viewer [29] software.

This study has been approved by the Medical Ethics Committee of Beijing Hospital of Traditional Chinese Medicine Affiliated to Capital Medical University (2023BL02-064-01) and Ethics Committee of Beijing University of Chinese Medicine (2024BZYLL0101). All participants were provided with a written consent form and recruited signed it prior to being into our study.

AD is the primary cause of dementia, accounting for 50% to 70% of all cases of dementia [30]. As the aging population expands, the prevalence of AD is expected to rise, particularly in low- and middle-income countries. AD has emerged as the fifth leading cause of mortality among individuals aged 65 years and older, serving as a direct contributor to dependency and disability [31]. AD is a neurodegenerative disorder characterized by the progressive and often irreversible degeneration of neurons, with treatment efficacy diminishing as the age of the patient increases, irrespective of the therapeutic interventions used [32]. According to the AD diagnostic guidelines published in the United States, the symptoms of aMCI fulfill the clinical criteria for the predementia stage [33]. Implementing early warning systems for aMCI and ensuring timely intervention during the initial stages of the disease can significantly enhance the prognosis of aMCI. Early screening and diagnosis are essential prerequisites for timely intervention. Given the insidious onset of aMCI, the methodologies used for screening and diagnosis hold critical significance in its management. This study uses screening and diagnostic methodologies that integrate clinical symptoms, localized brain alterations, pathological characteristics, and noninvasive yet convenient detection techniques to deliver a more comprehensive diagnosis and evaluation of the disease from multiple perspectives. While the symptoms of aMCI do not significantly disrupt an individual’s routine daily activities, they are associated with objectively lower performance on cognitive neuropsychological assessments when compared to individuals diagnosed with AD [34]. The clinical manifestations of aMCI can be primarily categorized into 3 domains: cognitive decline, mild deficits in complex work-related daily activities, and noncognitive psychiatric symptoms. The MoCA encompasses a wide array of cognitive domains, enabling the evaluation of an individual’s overall cognitive functioning. The Functional Activities Questionnaire evaluates intricate social competencies and daily living activities. Apathy, depression, anxiety, and atypical nocturnal sleep behaviors are prevalent neuropsychiatric manifestations in individuals with aMCI. To assess these symptoms, the Hamilton Anxiety Scale and the Depression Scale were used. Alterations in brain structure and function represent a significant contributor to aMCI. Imaging modalities offer noninvasive methodologies for assessing the brain’s structural and functional attributes, enabling an analysis of the interplay between these changes. Meanwhile, imaging phenotyping can provide a more accurate and objective basis for the classification and quantification of the disease, further aiding in the search for noninvasive imaging biomarkers for the early diagnosis of aMCI. The detection of biological markers in blood represents a noninvasive and cost-effective approach, with alterations in the levels of associated pathological proteins in plasma closely linked to structural changes in the brain and overall cognitive dysfunction. Furthermore, this method holds significant promise for predicting the transition from normal cognition to MCI [25]. Therefore, this trial used gold acupuncture needles as the primary intervention and used the “adjusting the spirit” acupuncture technique to address aMCI. The study also assessed the efficacy and safety of acupuncture treatment for aMCI through various evaluation methods conducted before and after the intervention. Simultaneously, by integrating imaging techniques with molecular biology, this study evaluates the effects of the Gold Needle “Regulating Spirit” method on the brains of patients with aMCI from multiple perspectives. It aims to deeply investigate the mechanisms underlying acupuncture intervention in aMCI and provide a theoretical foundation for the broader clinical application of Gold Needle “Regulating Spirit” treatment for aMCI, while also identifying potential biomarkers for early clinical diagnosis.

The “regulating spirit” technique, which primarily uses gold acupuncture needles as its therapeutic instrument, uses gold as the principal material. Therapeutic interventions are applied at the Baihui (GV20), Sishencong (EX-HN1), Shenting (GV24), Benshen (GB13), Yintang (GV29), Renzhong (GV26), Shenmen (HT7), Feishu (BL13), Xinshu (BL15), Ganshu (BL18), Pishu (BL20), Shenshu (BL23), and Geshu (BL17). The selection of acupuncture points is grounded in the principles of traditional Chinese medicine. The synergistic application of these acupoints facilitates cognitive clarity, enhances memory retention, and promotes the nourishment of internal organs [35]. The acupoints selected for this trial were determined based on the principles of Diagnosis and Treatment Based on Overall Analysis of the Illness and the Patient’s Condition in traditional Chinese medicine, as well as the clinical practice guidelines established in China. Nevertheless, the scientific evidence supporting these perspectives remains limited, and there is a notable deficiency of high-quality research, resulting in ambiguity regarding the findings—particularly concerning whether the therapeutic effects of gold acupuncture surpass those of conventional acupuncture. This was substantiated through a RCT, establishing a basis for subsequent large-scale RCTs. Due to the inherent characteristics of acupuncture treatment, implementing a blind method poses significant challenges; therefore, we will use a blinded approach to categorize participants into the Gold Needle “Regulating Spirit” group, the general acupuncture “Regulating Spirit” group, and the sham acupuncture group. Considering the placebo effect associated with sham acupuncture, a healthy cohort was designated as the negative control group. The primary challenge of this study lies in the fact that patients with aMCI are typically older, and the manifestations of aMCI tend to be quite subtle, rendering them less conspicuous within the community and not highly regarded by the general population. Moreover, the duration of the intervention for participants with aMCI in this trial was extended, making it crucial to assess their willingness to adhere to treatment throughout the study and their cooperation in the collection of various outcome measures.

We anticipate that by the end of the trial, we will be able to definitively ascertain whether the gold acupuncture needle technique for “regulating spirit” offers a significant advantage in treating aMCI, and further investigate the nature of this therapeutic benefit, to offer a more efficacious intervention for the clinical management of aMCI and the prevention of AD.