[Application prospect of conditional gene editing animals and viral vectors in experimental research of acupuncturology].

Conditional gene editing animals and viral vectors have been widely applied in the research fields of biology and medicine. Recently, they are also used as the effective approaches to reveal the underlying mechanism of acupuncture from the nervous system to the specific molecules. In order to further understand the application of conditional gene editing animals and viral vectors, in this article, we analyze their characteristics, advantages and recent development in the field of acupuncture research and discuss their potential roles and prospect in the future.

Sensory and autonomic innervation of the local tissues at traditional acupuncture point locations GB14, ST2 and ST6.

OBJECTIVE: To visualize and compare the sensory and autonomic innervation of the local tissues at the sites of different traditional acupuncture points in the rat forehead and face by histochemical examination. METHODS: GB14 (Yangbai), ST2 (Sibai) and ST6 (Jiache) were selected as the representative traditional acupuncture points in this study, and the local tissues at these sites were dissected in rats after perfusion followed by double or triple fluorescent histochemical staining. Here, calcitonin gene-related peptide (CGRP), tyrosine hydroxylase (TH) and vesicular acetylcholine transporter (VAChT) were used to label the sensory, sympathetic and parasympathetic nerve fibers, respectively. RESULTS: The CGRP+ sensory, TH+ sympathetic and VAChT+ parasympathetic nerve fibers were simultaneously demonstrated in the local tissues at GB14, ST2 and ST6. Although the three kinds of nerve fibers ran in parallel or intermingled with each other, by the analysis from the view of three-dimensional reconstruction, it was clear that each of them distributed in an independent pattern to their corresponding target tissues including the blood vessels, hair follicles, arrector pili and subcutaneous muscles, as well as sebaceous glands. CONCLUSION: Our study demonstrated the sensory and autonomic innervation of the local tissues at GB14, ST2 and ST6, providing neurochemical evidence indicating that the CGRP+ sensory, TH+ sympathetic and VAChT+ parasympathetic nerve fibers form a neural network at these point locations that may respond to acupuncture stimulation.

Temporal alterations in pericytes at the acute phase of ischemia/reperfusion in the mouse brain.

Pericytes, as the mural cells surrounding the microvasculature, play a critical role in the regulation of microcirculation; however, how these cells respond to ischemic stroke remains unclear. To determine the temporal alterations in pericytes after ischemia/reperfusion, we used the 1-hour middle cerebral artery occlusion model, which was examined at 2, 12, and 24 hours after reperfusion. Our results showed that in the reperfused regions, the cerebral blood flow decreased and the infarct volume increased with time. Furthermore, the pericytes in the infarct regions contracted and acted on the vascular endothelial cells within 24 hours after reperfusion. These effects may result in incomplete microcirculation reperfusion and a gradual worsening trend with time in the acute phase. These findings provide strong evidence for explaining the "no-reflow" phenomenon that occurs after recanalization in clinical practice.

Alexa Fluor 488-conjugated cholera toxin subunit B optimally labels neurons 3-7 days after injection into the rat gastrocnemius muscle.

Neural tract tracing is used to study neural pathways and evaluate neuronal regeneration following nerve injuries. However, it is not always clear which tracer should be used to yield optimal results. In this study, we examined the use of Alexa Fluor 488-conjugated cholera toxin subunit B (AF488-CTB). This was injected into the gastrocnemius muscle of rats, and it was found that motor, sensory, and sympathetic neurons were labeled in the spinal ventral horn, dorsal root ganglia, and sympathetic chain, respectively. Similar results were obtained when we injected AF594-CTB into the tibialis anterior muscle. The morphology and number of neurons were evaluated at different time points following the AF488-CTB injection. It was found that labeled motor and sensory neurons could be observed 12 hours post-injection. The intensity was found to increase over time, and the morphology appeared clear and complete 3-7 days post-injection, with clearly distinguishable motor neuron axons and dendrites. However, 14 days after the injection, the quality of the images decreased and the neurons appeared blurred and incomplete. Nissl and immunohistochemical staining showed that the AF488-CTB-labeled neurons retained normal neurochemical and morphological features, and the surrounding microglia were also found to be unaltered. Overall, these results imply that the cholera toxin subunit B, whether unconjugated or conjugated with Alexa Fluor, is effective for retrograde tracing in muscular tissues and that it would also be suitable for evaluating the regeneration or degeneration of injured nerves.

Histochemistry of microinfarcts in the mouse brain after injection of fluorescent microspheres into the common carotid artery.

The mouse model of multiple cerebral infarctions, established by injecting fluorescent microspheres into the common carotid artery, is a recent development in animal models of cerebral ischemia. To investigate its effectiveness, mouse models of cerebral infarction were created by injecting fluorescent microspheres, 45-53 µm in diameter, into the common carotid artery. Six hours after modeling, fluorescent microspheres were observed directly through a fluorescence stereomicroscope, both on the brain surface and in brain sections. Changes in blood vessels, neurons and glial cells associated with microinfarcts were examined using fluorescence histochemistry and immunohistochemistry. The microspheres were distributed mainly in the cerebral cortex, striatum and hippocampus ipsilateral to the side of injection. Microinfarcts were found in the brain regions where the fluorescent microspheres were present. Here the lodged microspheres induced vascular and neuronal injury and the activation of astroglia and microglia. These histopathological changes indicate that this animal model of multiple cerebral infarctions effectively simulates the changes of various cell types observed in multifocal microinfarcts. This model is an effective, additional tool to study the pathogenesis of ischemic stroke and could be used to evaluate therapeutic interventions. This study was approved by the Animal Ethics Committee of the Institute of Acupuncture and Moxibustion, China Academy of Chinese Medical Sciences (approval No. D2021-03-16-1) on March 16, 2021.

[Application of Adobe Illustrator for drawing schematic diagrams of anatomical structure of acupoints -- taking Five Shu-points of Jueyin Meridian as an example].

The schematic diagram is an indispensable part of research article in life science，which can effectively and intuitively show the specific content of the article with simple composition. As an intuitive expression of the interdisciplinary subject at the early stage, the schematic diagram in the literature has gained an all-round improvement in the quality along with the deve-lopment of computer technology and modern drawing tools. In contrast，the level of the schematic diagram appears to be lagged behind in the field of acupuncture research papers. In order to improve the drawing level of this field, we took the Five Shu-points of Hand and Foot Jueyin Meridian as an example, and drew some vector diagrams of their anatomical structures including the body surface, skeleton, nerves, arteries and veins by using the Adobe Illustrator image software, through which we sum up a simple and easy-to-learn process including application skills and key points needing attention. We hope these methods can play a role in the acupuncture research in the future.

[Characteristics of "deqi" and myoelectricity in different tissue structures of acupoint].

OBJECTIVE: To investigate the effect of acupuncture in different tissue structures on deqi and the electromyography of acupoint area. METHODS: Twenty healthy subjects, respectively accepted 4 kinds of needling stimulation, i.e. stimulating skin at Zusanli (ST36), stimulating ST36 with and without skin anesthesia using compound lidocaine cream, and stimulating at Dubi (ST35) without skin anesthesia. Deqi sensation of the acupuncturist and subjects were measured according to MGH Acupuncture Sensation Scale (MASS) during needling, and the myoelectricity around the acupoints was recorded simultaneously. The occurrence rate and intensity of the different deqi sensations, the relationship between the acupuncturist's and subjects' deqi sensations, and the integrated electromyogram (iEMG) were analyzed. RESULTS: Sharp pain and tingling were the main sensations during skin needling at ST36. Fullness, dull pain, soreness and acupuncturist's tightness were the main sensations during needling with or without skin anesthesia at ST36. Fullness was the main sensation during needling at ST35, while the intensity was lower than that during needling at ST36. A positive correlation in the intensity was found between subjects' fullness and acupuncturist's tightness during needling with or without skin anesthesia at ST36. The subjects' fullness appeared earlier about 5 seconds than acupuncturist's tightness. The iEMGs during subjects' fullness and acupuncturist's tightness were 2-3 times of that before needling. CONCLUSION: Deqi sensations such as subjects' fullness, dull pain, soreness and acupuncturist's tightness are mainly related to the activity of the muscles under the acupoints. Subjects' fullness and acupuncturist's tightness always appear together. Acupuncturist's tightness may be mediated by the muscle stretch reflex induced by needling stimulation.

Effects of Three Needling Manipulations of Zusanli (ST 36) on Deqi Sensations and Surface Myoelectricity in Healthy Participants.

OBJECTIVE: To investigate the effects of different acupuncture manipulations on Deqi sensations and surface myoelectricity, and explore the correlation between Deqi sensations and needling manipulations. METHODS: Forty-five healthy participants accepted twirling, lifting-thrusting, and twirling plus lifting-thrusting manipulanions at right Zusanli (ST 36), respectively. The acupuncturist's and participants' Deqi sensations were collected by MGH Acupuncture Sensation Scale (MASS). The intensity and occurrence rate of soreness, dull pain, pressure, heaviness, fullness, numbness, sharp pain, warmth, coolness, and throbbing feelings of participants, and tightness, smooth, and tangle feelings of acupuncturist were measured. The correlation between the acupuncturist's and participant's Deqi sensations was analyzed. Surface electromyogram (EMG) were recorded before, during and after needling in 30 participants. The integrated EMG (iEMG), mean power frequency (MPF) and media frequency (MF) were analyzed. RESULT: Both fullness and soreness of participants and tightness of acupuncturist were the most frequently occurred ones. A positive correlation between participants' fullness and acupuncturist's tightness was observed during the three aforementioned needling manipulations (P<0.05, OR>1). Almost all the needling sensations measured in the present study could be induced by the three needling manipulations. However, strength of Deqi sensations was exhibited as lifting-thrusting > twirling plus lifting-thrusting > twirling according to MASS index. The iEMG values were increased and MPF, MF values were decreased during needling compaired to those before needling, especially during lifting-thrusting (P<0.01). CONCLUSIONS: The intensity and occurrence rate of the different Deqi sensations induced by different needling manipulations were basically similar. The fullness and soreness were both the most frequently induced Deqi sensations. The strongest Deqi sensation could be induced by lifting-thrusting manipulation. There is a positive correlation between participants' fullness and acupuncturist's tightness during the three needling manipulations. The myoelectricity around the acupoint is related to Deqi responses. (Registration No. AMCTR-IOR-20000314).

[Microstructural characteristics of lymphatic vessels in skin tissues of acupoints "Taichong" and "Yongquan" in the rat].

OBJECTIVE: To provide a new method for investigating the histological characteristics of acupoints by obser-ving the microstructure of the lymphatic vessels in the skin tissue of "Taichong" (LR3) and "Yongquan" (KI1) regions. METHODS: Six male SD rats were used in the present study. The skin tissue of LR3 and KI1 from the hind foot were taken following transcardial perfusion with 4% paraformaldehyde. The skin tissues were cut into sagittal sections with a freezing microtome and stained by fluorescent immunohistochemistry with lymphatic vessel endothelial hyaluronan receptor 1 (LYVE-1), calcitonin gene-related peptide (CGRP), and phalloidin for displaying the lymphatic vessels, nerve fibers, and blood vessels, separately. The samples were viewed and recorded using fluorescent microscope and laser scanning confocal microscope. RESULTS: In the skin tissue of LR3 and KI1 regions, the lymphatic vessels, nerve fibers, and blood vessels were labeled with LYVE-1, CGRP and phalloidin, respectively. The lymphatic capillaries were found to start from the enlarged blind end and distribute in the dermis and subcutaneous tissues with various forms, crisscrossing. Abundant blood capillaries at various thickness distributed around the lymphatic capillaries in a parallel or crossed pattern, intermingled with free nerve fibers. CONCLUSION: The lymphatic capillaries, blood capillaries and nerve fibers extensively distribute in the skin tissues of LR3 and KI1 regions in rats, suggesting an involvement of the immunomodulation in the effects of acupuncture in pathological conditions, despite being not limited to the acupoint regions in the distribution of lymphatic capillaries.

Antihypertensive and Antifibrosis Effects of Acupuncture at PC6 Acupoints in Spontaneously Hypertensive Rats and the Underlying Mechanisms.

Long-term hypertension can lead to both structural and functional impairments of the myocardium. Reversing left ventricular (LV) myocardial fibrosis has been considered as a key goal for curing chronic hypertension and has been a hot field of research in recent years. The aim of the present work is to investigate the effect of electroacupuncture (EA) at PC6 on hypertension-induced myocardial fibrosis in spontaneously hypertensive rats (SHRs). Thirty SHRs were randomized into model, SHR + EA, and SHR + Sham EA groups with WKY rats as a normal control. EA was applied once a day for 8 consecutive weeks. The cardiac fibrosis as well as the underlying mechanisms were investigated. After 8 weeks of EA treatment at PC6, the enhanced myocardial fibrosis in SHRs was characterized by an increased ratio of left ventricular mass index (LVMI), collagen volume fraction (CVF), and elevated content of hydroxyproline (Hyp) as well as the upregulated expression of collagen I and collagen III in myocardium tissue of SHRs. All these abnormal alterations in the SHR + EA group were significantly lower compared to the model group. In addition, EA at PC6 significantly improved the pathological changes of myocardial morphology. Meanwhile, the increased levels of angiotensin II (Ang II) and tumor necrosis factor-α (TNFα) and expression of transforming growth factor ß1 (TGF-ß1), connective tissue growth factor (CTGF), matrix metalloproteinase (MMP)-2, and MMP-9 in the serum or heart tissue of SHRs were also markedly diminished by EA. These results suggest that EA at bilateral PC6 could ameliorate cardiac fibrosis in SHRs, which might be mediated by the regulation of the Ang II - TGF-ß1 pathway.

[How to improve the quality of histological pictures in the manuscripts of Acupuncture Research].

Histological pictures are important parts of medical research articles, and a good histological picture not only visualizes the content of the article, but also makes it easier for readers to understand. With the rapid development of microscopic technology and image processing technology, journals have higher requirements for the quality of histological pictures and more strict requirements for the editing and annotation of pictures. Based on an analysis of the current status of histological pictures in the articles of Acupuncture Research, this article proposes our suggestions and Methods for rational use of histological pictures, so as to provide a reference for improving the quality of histological pictures in the articles published in Acupuncture Research.

GABAergic Inhibition of Spinal Cord Dorsal Horns Contributes to Analgesic Effect of Electroacupuncture in Incisional Neck Pain Rats.

BACKGROUND: Acupuncture has shown to be effective in relieving post-surgical pain. Nonetheless, its underlying mechanisms remain largely unknown. In the present study, we investigated the effect of electroacupuncture (EA) on the expression of GABA, GABA-A receptor (R) and GABA-BR in the spinal cord dorsal horns (DHs), and the involved neural cells in rats with incisional neck pain. MATERIALS AND METHODS: Male SD rats were randomly divided into control, model, Futu (LI18), Hegu-Neiguan (LI4-PC6), and Zusanli-Yanglingquan (ST36-GB34) groups. The incisional neck pain model was established by making a longitudinal incision and repeated mechanical separation along the thyroid gland region. EA (2Hz/100Hz, 1mA) was applied to LI18, LI4-PC6, ST36-GB34 separately for 30min, once at 4, 24 and 48h after incision. The local thermal pain threshold (TPT) of the focus was measured and the expression of GABA, and GABAR proteins and mRNAs detected by immunofluorescence stain and quantitative RT-PCR, respectively. RESULTS: The analgesic effect of LI18 and LI4-PC6 was superior to that of ST36-GB34 in incisional neck pain rats. Moreover, the EA stimulation of LI18 or LI4-PC6 increased the expression of GABA and GABA-Aα2 and GABA-Aß3, GABA-B1, and GABA-B2 mRNAs in spinal DHs 4h after surgery, while GABA-A and GABA-B antagonists inhibited the analgesic effect of LI18. Immunofluorescence double staining showed that GABA was expressed on astrocytes and neurons, and GABA-B expressed only on neurons. CONCLUSION: EA of both LI18 and LI4-PC6 has a good analgesic effect in incisional neck pain rats, which is closely related to their effects in upregulating the expression of GABA and its receptors in spinal DHs. The effects of LI18 and LI4-PC6 EA are obviously better that those of ST36-GB34 EA, and GABA is expressed on neurons and astrocytes.

Response to Commentary on "The neuroinvasive potential of SARS-CoV-2 may play a role in the respiratory failure of COVID-19 patients".

In a recent review, we have suggested a neuroinvasive potential of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and its possible role in the causation of acute respiratory failure of coronavirus disease 2019 (COVID-19) patients (J Med Viol doi: 10.1002/jmv.25728), based upon the clinical and experimental data available on the past SARS-CoV-1 and the recent SARS-CoV-2 pandemic. In this article, we provide new evidence recently reported regarding the neurotropic potential of SARS-CoV-2 and respond to several comments on our previously published article. In addition, we also discuss the peculiar manifestations of respiratory failure in COVID-19 patients and the possible involvement of nervous system.

The neuroinvasive potential of SARS-CoV2 may play a role in the respiratory failure of COVID-19 patients.

Following the severe acute respiratory syndrome coronavirus (SARS-CoV) and Middle East respiratory syndrome coronavirus (MERS-CoV), another highly pathogenic coronavirus named SARS-CoV-2 (previously known as 2019-nCoV) emerged in December 2019 in Wuhan, China, and rapidly spreads around the world. This virus shares highly homological sequence with SARS-CoV, and causes acute, highly lethal pneumonia coronavirus disease 2019 (COVID-19) with clinical symptoms similar to those reported for SARS-CoV and MERS-CoV. The most characteristic symptom of patients with COVID-19 is respiratory distress, and most of the patients admitted to the intensive care could not breathe spontaneously. Additionally, some patients with COVID-19 also showed neurologic signs, such as headache, nausea, and vomiting. Increasing evidence shows that coronaviruses are not always confined to the respiratory tract and that they may also invade the central nervous system inducing neurological diseases. The infection of SARS-CoV has been reported in the brains from both patients and experimental animals, where the brainstem was heavily infected. Furthermore, some coronaviruses have been demonstrated able to spread via a synapse-connected route to the medullary cardiorespiratory center from the mechanoreceptors and chemoreceptors in the lung and lower respiratory airways. Considering the high similarity between SARS-CoV and SARS-CoV2, it remains to make clear whether the potential invasion of SARS-CoV2 is partially responsible for the acute respiratory failure of patients with COVID-19. Awareness of this may have a guiding significance for the prevention and treatment of the SARS-CoV-2-induced respiratory failure.

[The shortcut pathway between scalp acupuncture and brain].

Scalp acupuncture is widely used in clinical practice, and there is still a lack of systematic studies on its nerve pathways and mechanism of action. Scalp acupoints are innervated by the trigeminal nerve which is also involved in the regulation of the leptomeninges and related arterioles nourishing the cerebral cortex. Animal experiments and human trials have shown that scalp acupuncture can exert an effect through the shortcut between the axon reflexes from the primary neurons in the trigeminal ganglion (its axon branches may separately innervate scalp acupoints and the leptomeninges directly regulating nutrient and active substances in brain tissue) and the secondary neurons of the trigeminal nucleus caudalis (head and face acupoints interact with afferents of brain tissue through presynaptic dorsal root reflex and postsynaptic neurogenic responses and convergence), which may have regulatory and therapeutic effects on intracranial tissue.

[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.

[Distribution of serotonin-positive cells in local cutaneous tissues of different acupuncture points at hindlimbs in rats].

OBJECTIVE: To investigate the expression of serotonin (5-HT) in local tissue of different acupoint regions at the hind limbs of rats. METHODS: A total of 6 male SD rats were used in the present study. The local tissues of "Neiting"(ST44), "Zusanli"(ST36) and "Futu"(ST32) at the hindlimbs were dissected after transcardiac perfusion with fixative solution containing 4% paraformaldehyde, phosphate buffer, etc., fixed again in 4% paraformaldehyde and cut into sections (at 20 µm thick). Then, the tissue sections were subjective into immunofluorescence stain for displaying 5-HT positive cells and calcitonin gene-related peptide (CGRP) -positive fibers, vessel-like structure (labelled with phalloidin) and cellular nucleus (labelled with DAPI), separately. After that, the tissue samples were observed and recorded using a laser confocal microscope. RESULTS: The 5-HT labeled cells presented flat or round form in the local tissues of the 3 acupoint regions. These cells were mainly located in the dermis and subcutaneous tissue, and concentrated around the vessel-like structure and CGRP-positive nerve fibers. The number of cutaneous 5-HT positive cells was most in ST44 (156.6±12.1), and fewer in ST36 (83.5±18.2)and ST32(53.3±5.2)in turn. CONCLUSION: 5-HT-positive cell is an important member in local tissue of acupoints (ST44, ST36 and ST32), and varies in number according to their locations, suggesting an involvement of different densities of 5-HT-positive cells in different levels of regulatory effects of acupuncture stimulation of these acupoints.

[Moxa-stick moxibustion activates mast cells of small intestine tissue in rats].

OBJECTIVE: To explore the effect of moxa-stick moxibustion and joss-stick moxibustion at "Guanyuan" (CV4) on the activity of mast cells in the small intestine tissue in rats. METHODS: Twelve male SD rats were randomly divided into control, joss-stick moxibustion and moxa-stick moxibustion groups (n=4 rats in each group). Joss-stick or moxa-stick moxibustion was applied to CV4 for 10 min. After moxibustion, the skin temperature of the CV4 region was measured immediately with a thermometer. The mast cells and nerve fibers in the small intestine tissue were displayed by immunofluorescence histochemistry. RESULTS: Compared with the control group, the skin temperature of the CV4 region in both the joss-stick and the moxa-stick groups were significantly increased (P<0.05), while the skin temperature of the moxa-stick group was significantly higher than that in the joss-stick group (P<0.05). There were a large number of tryptase-positive mast cells in the small intestine of rats, some of which were co-expressed with lysosomal-associated membrane protein 1, displaying an activated state. The average numbers of mast cells in the control, joss-stick and moxa-stick groups were 9.2±3.6, 10.8±5.3 and 17.1±6.3, respectively, being significantly higher in the moxa-stick group than in the control and joss-stick groups (P<0.05). In addition, calcitonin gene-related peptide(CGRP) and neuropeptide Y(NPY) positive nerve fibers were found around the mast cells in the small intestinal tissues. CONCLUSION: Moxa-stick moxibustion gives rise a higher temperature at CV4 to activate mast cells surrounded by CGRP and NPY positive nerve fibers in the small intestine tissue in rats, suggesting an involvement of the sensory and sympathetic nervous system in the activation of intestinal mast cells possibly by way of somatic sympathetic reflex.

[Relationship between acupuncture sensations of deqiand different organizational structures of acupoint area].

Acupuncture sensation, as one of the essential signs of deqi (arrival of qi), is closely associated with curative effect. With systematically reviewing the literature regarding relationship between acupuncture sensations of deqi and different organizational structures of acupoint area (skin, connective tissue, muscles, nerves and blood vessels), it is believed that acupuncture sensation of deqi is the product of various organizational structures in acupoint area, which is closely related to the nervous system. The deqi sensations sensed by the hands of acupuncturists, which is called sense of needle stagnation, maybe caused by the contraction of muscles in the acupoint area during the process of needle manipulation. By system review, this paper puts forward a new research idea to explore the role of different manipulations of needling in generating the sense of qi-generating needle based on different acupoint structure and brain function response system, which could provide some reference for clarifying the scientific mechanism and clinical application of acupuncture sensations of deqi.

[From molecular pharmacy to electroceuticals: SPARC program and acupuncture research].

The NIH-funded "Stimulating Peripheral Activity to Relieve Conditions (SPARC)" program has been initiated in Octomber 2016, aiming at developing high resolution neural circuit maps and next generation neural modulation devices. This program has brought great stimulus to acupuncturists and acupuncture researchers both at home and abroad. Is the SPARC program a driving force or a challenge of acupuncture research? In the present study, we introduced the SPARC program and compared it with current acupuncture research. The first step of SPARC is to better map neural circuits associated with disease on the anatomical level so as to identify the best points for intervention, and to decode the neural language at these intervention points for developing a dictionary of patterns associated with health and disease states on the signaling level. Similarly, our acupuncture research also focuses on revealing the neural encoding of acupuncture stimulation and its effect on visceral function, seeking suitable stimulation parameters to regulate the abnormal visceral activity precisely. Therefore, the common point of SPARC program and acupuncture research is the scientific basis of peripheral somatic neuronal regulation, and their difference is that acupuncture regulates the visceral function through multiple neural circuits and neural feedbacks by stimulating the sensitized points or acupoints, achieving homeostasis at last. Acupuncture-induced regulation effect is limited and the therapy is safe. Whereas, "stimulating periphe-ral activity (SPA)" can regulate the visceral organs precisely but without neural feedback. Inevitably, SPA will produce tolerance or side effects. Therefore, there is still much work to be done in terms of the initiation of trigger stimulation and the feedback inhibition of target organ effects. The SPARC program is definitely a powerful force in revealing the mechanisms by which acupuncture works.