Effectiveness and cerebral responses of multi-points acupuncture for primary insomnia: a preliminary randomized clinical trial and fMRI study.

BACKGROUND: Primary insomnia (PI) is characterized by difficulties in initiating sleep or maintaining sleep, which lead to many serious diseases. Acupuncture for PI has drawn attention with its effectiveness and safety. However, the operation of choosing acupoints lacks scientific suggestion. Our trial aims to provide reference and scientific basis for the selection of acupoints and to explore its possible mechanism. METHODS: A patient-assessor-blinded, randomized and sham controlled trial was designed to compare the efficacy of 5-weeks acupuncture at a single acupoint, the combination of multi-acupoints, and a sham point. The Pittsburgh sleep quality index and Athens Insomnia Scale questionnaire were used for the primary clinical outcomes, while polysomnography was performed for the secondary clinical outcomes. The resting state functional MRI was employed to detect the cerebral responses to acupuncture. The brain activity in resting state was measured by calculating the fractional amplitude of low-frequency fluctuations (fALFF), which reflected the idiopathic activity level of neurons in the resting state. These results were analyzed by two factorial ANOVA test and post-hoc t-tests. RESULTS: The clinical outcomes suggest that acupuncture could improve clinical symptoms, and the combination of multi-acupoints might lead to a better clinical efficacy. The rs-fMRI results suggested that the brain activity of certain regions was related to the sleep experience, and acupuncture could regulate the activity of these regions. Furthermore, the combination of multi-acupoints could impact more regions which were influenced by the sleep experience. CONCLUSIONS: Acupuncture has been proven to be beneficial for PI patients, and the combination of multi-acupoints might improve its efficacy. TRIAL REGISTRATION: This trial has been registered on the U.S. National Library of Medicine (https://clinicaltrials.gov) ClinicalTrials.gov Identifier: NCT02448602 . Registered date: 14/04/2015.

Efficacy of compatible acupoints and single acupoint versus sham acupuncture for functional dyspepsia: study protocol for a randomized controlled trial.

BACKGROUND: Acupoint selection is a key factor in the treatment of diseases and has not been well studied. The aim of this trial is to explore the differences in efficacy between compatible acupoints and a single acupoint for patients with functional dyspepsia (FD). METHODS: This randomized controlled trial will be conducted in the First Affiliated Hospital of Changchun University of Chinese Medicine in China. Two hundred and sixteen FD patients will be randomly assigned to the compatible acupoints group, single acupoint group, or sham acupuncture group. This trial will include a 1-week baseline period, a 4-week treatment period, and a 4-week follow-up period. During the 4-week treatment period, patients will receive 20 sessions of acupuncture (weekly cycles of one session per day for 5 consecutive days followed by a 2-day break). The primary outcome will be a change in the Nepean Dyspepsia Life Quality Index from baseline to after the 4-week treatment period. Secondary outcome measures will include the dyspeptic symptom sum score, Overall Treatment Effect questionnaire, and 36-item Short Form survey. Adverse events also will be recorded. Ultraweak photon emission and metabolomics tests will be performed at baseline and at the end of treatment to explore the mechanisms of the differences between compatible acupoints and a single acupoint. DISCUSSION: The results of this trial will allow us to compare the difference in efficacy between compatible acupoints and a single acupoint. The findings from this trial will be published in peer-reviewed journals. TRIAL REGISTRATION: Acupuncture-Moxibustion Clinical Trial Registry, AMCTR-IPC-18000176, registered on 4 March 2019; Chinese Clinical Trial Registry, ChiCTR1900023983, registered on 23 June 2019.

[Review on application of neural tracing technique to experimental research of acupuncture].

OBJECTIVE: To collect literatures on the application of neural tracing technique in experimental acupuncture research, and summarize its application status in acupoints, meridians and Zang-fu organs. METHODS: We collected papers published from databases of CNKI (1979-2017), Wan-Fang (1990-2017), VIP (1989-2017) and PubMed(1997-2017)by using keywords of "neural tracing technique" "neuroanatomic tracing" "neural tracers" and "acupuncture" "electroacupuncture" "auricular acupuncture" "eye acupuncture" "meridians" "acupuncture points" "acupoint injection", and made a summary about the current state of application of neural tracing technique in the fields of acupoint, meridian and Zang-fu organs (viscera). RESULTS: A total of 94 articles were collected, the most commonly used neural tracers were horseradish peroxidase and cholera toxin subunit B. The experimental animals used were rat, rabbit, cat, monkey, etc., and injection site was acupoint. After the injection of neural tracers, the survival time of animals was range from 1 day to 12 weeks, and the labelled tissues included neurons and nerve fibers of the sensory, motor, and autonomic systems. The outcomes of neural labeling mainly revealed the segmental pattern, neuroanatomical connection (neural pathways/circuits) and chemical features (shown by immunohistochemical staining) of neurons and nerve fibers innervating both the acupoints and visceral organs, suggesting their involvement in the effect of acupuncture and moxibustion treatment. CONCLUSION: This application of neural tracing technology help us understand the under-lying mechanisms of acupuncture and moxibustion interventions from different perspectives of neural pathways/circuits and related chemical properties, which also lays a greater role for this technology in future experimental acupuncture research.

[Analysis of Compatibility Laws for Acupoint Selection of Acupuncture in Treating Diabetic Gastroparasis].

OBJECTIVE: To summarize and analyze the compatibility laws for acupoint selection of acupuncture in treating diabetic gastroparasis (DGP) in modern literatures of clinical researches. METHODS: Retrieved were literatures related to treating DGP by acupuncture or acupuncture combined other therapies from PubMed, CNKI, and WF from 1982 to 2014. Analyzed were frequency of acupoint use, meridians selected acupoints belonged to, regions selected, association laws of selected compatible acupoints. RESULTS: Retrieved were 35 with compatibility frequencies more than 15 listed as follows: compatibility frequency of Zusanli (ST36) and Zhongwan (CV12) was 33 (94.29%); compatibility frequency of Zusanli (ST36) and Neiguan (PC6) was 23 (65.71%), compatibility frequency of Zhongwan (CV12) and Neiguan (PC6) was 22 (62.86%), compatibility frequency of Zusanli (ST36), Zhongwan (CV12), and Neiguan (PC6) was 22 (62.86%); compatibility frequency of Zusanli (ST36) and Sanyinjiao (SP6) was 16 (45.71%); compatibility frequency of Zhongwan (CV12) and Sanyinjiao (SP6) was 16 (45.71%); compatibility frequency of Zusanli (ST36), Zhongwan (CV12), and Sanyinjiao (SP6) was 15 (42.86%). Meridians selected acupoints belonged to were sequenced as Foot Yangming Stomach channel, Ren channel, Foot Taiyang Bladder channel, and so on. Acupoints selected were mainly in lower limbs, chest and abdomen, waist and back. The compatibility of Zusanli (ST36) and Zhongwan (CV12) was the most often used with the highest frequency. CONCLUSIONS: The compatibility laws for acupoint selection of acupuncture in treating DGP were mainly dominated as upper-lower selection, three regions selection, local selection, anterior-posterior selection. The compatibility laws for acupoint selection of acupuncture along meridians were mainly dominated as the convergence points and exterior-interior meridian points.

[The expression of calcitonin gene-related peptide in the sensory and motor neurons associated with "Hegu" (LI 4) in the rat].

OBJECTIVE: To investigate the chemical expression of the primary sensory and motor neurons associated with "Hegu" (LI 4) area by using neural tracing and fluorescent immunohistochemical techniques. METHODS: A total of 4 SD rats were used in the present study. After anesthesia, the rats received microinjection of 5 microL of Alexa Fluor 594 conjugated cholera toxin subunit B (AF 594-CTB) into the right "Hegu" (LI 4) area for observing the distribution of the related primary sensory neurons in dorsal root ganglia (DRGs) and motor neurons in the spinal cord (C 1 - C 5). Three days after the microinjection, the rats were anesthetized and transcardiacally perfused with 4% paraformaldehyde, followed by routine section of the DRGs and spinal cord tissues. The labeled neurons were observed by light microscope and their chemical characteristics determined by calcitonin gene-related peptide (CGRP) fluorescent immunohistochemical method. RESULTS: All the AF 594-CTB labeled neurons in DRGs and spinal cord were located on the side of the tracer injection. The labeled sensory neurons distributed in DRGs of cervical (C) 5 to thoracic (T) 1 segments with a high concentration in C 7 DRG in which 73.5% sensory neurons were shown to be CGRP-positive. The labeled motor neurons were found to be in the dorsolateral region of the spinal anterior horns from C 6 to T 1 segments, with a high concentration at C 8 in which 100% motor neurons were shown to be CGRP-positive. CONCLUSION: Acupoint LI 4 is innervated by sensory neurons from DRGs of C 5 - T 1 and motor neurons from anterior horns of the same segmental spinal cord in rats. Among them, 73.5% of the sensory neurons and 100% of the motor neurons are CGRP-positive, suggesting an involvement of CGRP in acupuncture stimulation-LI 4 induced therapeutic effects.

[Segmental and regional distribution of neurons and their axonal projection associated with acupoint "yongquan" (KI 1) in the rat: cholera toxin subunit B method].

OBJECTIVE: To observe the distribution of neurons and their axonal projection associated with "Yongquan" (KI 1) area by neural tracing technique with cholera toxin subunit B (CTB). METHODS: A total of 5 microL of 1% CTB solution was injected into the front central part of hind foot plantar (corresponding to the KI 1 region in the human body) using a Hamilton microsyringe. After 3 survival days, the rats were deeply anesthetized and transcardically perfused for collecting the dorsal root ganglia (DRGs), spinal cord and brain tissues. Following fixing in 4% paraforldehyde containing phosphate buffer solution (PBS) and incubation in PBS containing 25% glucose solution, the aforementioned tissues were sectioned to be stained with immunofluorescence or immunohistochemistry for revealing the labeled sensory neurons and their tansganglionic projection and motor neurons. RESULTS: All the labeled neurons (sensory and motor neurons) and transganglionic axonal projection appeared ipsilaterally to the injection side. The labeled sensory neurons were located in the DRGs of lumbar 3-5 segments (L 3-L 5) with a higher concentration at L 4, while motor neurons distributed in the dorsolateral portion of spinal ventral horn from L 3 to L 6 with a higher concentration at L 5. In addition, transganglionic axonal projections were found to situate in the medial part of laminae III - IV from L 3 to L 5, as far as in the gracile nucleus. CONCLUSION: Acupoint KI 1 area is innervated by sensory neurons in L 3-L 5 DRGs and motor neurons from the dorsolateral ventral horns of L 3-L 6. The axonal projection of the primary sensory neurons distributes in the medial part of laminae III - IV of L 3-L 5 and gracile nucleus, respectively.

Neuroanatomical basis for acupuncture point PC8 in the rat: neural tracing study with cholera toxin subunit B.

OBJECTIVES: This study was performed to investigate the innervations related to acupuncture point PC8 in rats using a neural tracing technique. METHODS: After 6 µL of 1% cholera toxin subunit B (CTB) was injected into the site between the second and third metacarpal bone in rats, a corresponding site to acupuncture point PC8 in the human body, CTB labelling was examined with immunofluorescence and immunohistochemistry in the dorsal root ganglia (DRG), spinal cord and brainstem. RESULTS: All CTB labelling appeared on the ipsilateral side of the injection. The labelled sensory neurons distributed from cervical (C)6 to thoracic (T)1 DRG, while the labelled motor neurons were located on the dorsolateral part of the spinal ventral horn ranging from the C6 to T1 segments. In addition, the transganglionically-labelled axonal terminals were found to be dense in the medial part of laminae 3-4 from C6 to the T1 spinal dorsal horn, as far as in the cuneate nucleus. CONCLUSIONS: These results indicate that sensory and motor neurons associated with PC8 distribute in a distinct segmental pattern. The sensory information from PC8 could be transganglionically transported to the spinal dorsal horn and cuneate nucleus.