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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.

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.

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.

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.