[Textual research on varieties of Draconis Sanguis].

This paper reviewed the historical evolution of the varieties of Draconis Sanguis in traditional Chinese medicine(TCM) and discussed several doubts. Draconis Sanguis used in ancient Europe and Arabia was derived from Dracaena plants, and that originating from Southeast Asia entered the market in the 16 th century. Draconis Sanguis was introduced into China in the 5 th century at the latest and was once mixed with shellac for use. Draconis Sanguis in the Tang Dynasty and before was the resin of Dracaena plants. Scholars in the Song Dynasty have known that Draconis Sanguis came from the resin of tall trees, but their understanding of origin plants was inconsistent with the facts. The origin of Draconis Sanguis in the Song Dynasty was basically determined to be Mirbat(Maliba), Cengtan, and Somali, as well as Socotra Archipelago. About 1371-1416, Draconis Sanguis prepared from Daemonorops draco was imported into China, and was recorded earlier in The Overall Survey of the Ocean's Shores(Ying Ya Sheng Lan) and Code of Great Ming Dynasty(Da Ming Hui Dian). Draconis Sanguis prepared from Dracaena plants was still authentic for a long time after the import of that from D. draco into China. During the Ming and Qing Dynasties, Dian Zhi(1625), a lost edition of Materia Medica in Southern Yunnan(Dian Nan Ben Cao), Textual Research on Reality and Titles of Plants(Zhi Wu Ming Shi Tu Kao), and other local chronicles recorded that a new type of Draconis Sanguis(Mu Xue Jie) was produced in Yuanjiang, Yunnan province. The New Yunnan Chronicles of the Republic of China recorded the production of another type of Draconis Sanguis(Qi Lin Jie) in Xishuangbanna. However, the authenticity of the above two types has been difficult to confirm. In modern times, Draconis Sanguis prepared from D. draco gradually became the mainstream variety. In the 1970 s, Dracaena cochinchinensi was found in Yunnan and other provinces, and Draconis Sanguis from D. cochinchinensi was developed. This study is expected to provide a solid and reliable literature support for the research and development of Draconis Sanguis, enrich historical materials, and provide new clues for follow-up research.

Paclitaxel, 5-fluorouracil, and leucovorin combination chemotherapy as first-line treatment in patients with advanced gastric cancer.

The aim of this retrospective analysis was to evaluate the efficacy and toxicity of combination chemotherapy with paclitaxel, 5-fluorouracil, and leucovorin (TFL) as first-line treatment in patients with advanced gastric cancer (AGC). One hundred and thirteen patients were enrolled in the study who were confirmed to have AGC by histopathology. These patients were treated with TFL: paclitaxel at a dose of 135 mg/m as a 3-h intravenous infusion on day 1, LV 400 mg/m as an intravenous infusion over 2 h on day 1, followed by 5-fluorouracil 2400 mg/m as an infusion over a 46-h period on 3 consecutive days. Cycles were repeated every 2 weeks. A total of 113 patients were assessed for their response to therapy. A total of three patients achieved complete responses and 46 patients achieved partial responses, yielding an overall objective response rate of 43.4% [95% confidence interval (CI): 34.3-52.5%]. Fifty-four cases of stable disease and 10 cases of progressive disease were observed in the remaining patients. The median time to progression and overall survival were 5.2 months (95% CI: 4.7-5.8 months) and 14.1 months (95% CI: 12.5-15.8 months), respectively. Toxicities were tolerable and moderate. The most common grade 3-4 toxicities included leukopenia (16.8%), neutropenia (17.7%), anemia (8.0%), thrombocytopenia (5.3%), and fatigue (6.2%). Combination chemotherapy with TFL offers an active and safe therapeutic approach for patients with AGC.

Promoting high-quality development of acupuncture and moxibustion. / 推进针灸学科的高质量发展.

In the past 20 years, the acupuncture-moxibustion discipline has made a great progress in clinical research, method construction, standard formulation, guideline promotion, basic theory and key scientific issue research. Internationally, the development of acupuncture and moxibustion has gradually begun to pay more attention to the basic issues of the discipline itself from focusing on clinical evidence. The National Institute of Health of USA pays close attention to the construction of acupoint knowledge base and database and to the transformation of peripheral nerve stimulation techniques, which brings forth opportunities and challenges for the development of acupuncture-moxibustion discipline. In the present paper, we analyze the shortcomings of the current development of acupuncture and moxibustion, put forward some strategies for high-quality development in the future, and sort out the basic scientific issues to form an academic consensus. We should employ modern scientific language to express the scientific connotations of the basic theory of acupuncture and moxibustion, and build an open and self-consistent modern theoretical system. In addition, we also should attract more multidisciplinary talents to harmoniously and assiduously work together, insist on continuous innovation to open up a new situation in the transformation of basic scientific research achievements, and establish a new theoretical system of "somato-medicine" represented by acupuncture and moxibustion. In this way, we will guide the acupuncture-moxibustion discipline to make an original contribution to the modern life science and future medicine.

Perspective of Calcium Imaging Technology Applied to Acupuncture Research.

Acupuncture, a therapeutic treatment defined as the insertion of needles into the body at specific points (ie, acupoints), has growing in popularity world-wide to treat various diseases effectively, especially acute and chronic pain. In parallel, interest in the physiological mechanisms underlying acupuncture analgesia, particularly the neural mechanisms have been increasing. Over the past decades, our understanding of how the central nervous system and peripheral nervous system process signals induced by acupuncture has developed rapidly by using electrophysiological methods. However, with the development of neuroscience, electrophysiology is being challenged by calcium imaging in view field, neuron population and visualization in vivo. Owing to the outstanding spatial resolution, the novel imaging approaches provide opportunities to enrich our knowledge about the neurophysiological mechanisms of acupuncture analgesia at subcellular, cellular, and circuit levels in combination with new labeling, genetic and circuit tracing techniques. Therefore, this review will introduce the principle and the method of calcium imaging applied to acupuncture research. We will also review the current findings in pain research using calcium imaging from in vitro to in vivo experiments and discuss the potential methodological considerations in studying acupuncture analgesia.

[Relationship between acupoint sensitization and changes in dorsal root ganglion neuron excitability in myocardial ischemia mice].

OBJECTIVE: To investigate the relationship between the sensitization state of acupoints on the surface of the myocardial ischemia (MI) model mice and the changes in the electrophysiological properties of the dorsal root ganglion (DRG) neurons in the corresponding spinal cord segment, and its underlying mechanism. METHODS: Sixty-eight male C57BL/6J mice were randomly divided into control and model groups (34 mice in each group). The model group received an intraperitoneal injection of 160 mg/kg isoproterenol (ISO) to establish the MI model, and the control group received an injection of the same dose of normal saline as the model group. After modeling for about 6 days, MI proportion was measured by HE staining to verify the pathological changes in the heart tissue. Evans blue (EB) dye was injected into the tail vein of mice to reflect the size, location, distribution, and number of exudates on the body surface. Then, whole-cell membrane currents, intrinsic excitability and membrane properties of different types of DRG neurons were evaluated by electrophysiological experiment in vitro. RESULTS: Compared with the control group, the heart size was larger, with pathological outcomes showing enlarged myocardial hypertrophy, destroyed structure of cardiomyocytes, with mononuclear cell infiltration among the cardiomyocytes in the model group. Compared with the control group, the number of EB exudation points was significantly increased (P<0.01), which were mainly concentrated in the epidermis near the T1-T5 segment of the spinal cord, "Feishu" (BL13), "Jueyinshu" (BL14) and "Xinshu" (BL15) in the model group. Compared with the control group, the rheobase and action potential amplitude (APA) of DRG medium-sized neurons were obviously decreased (P<0.01, P<0.05), while the whole-cell membrane currents, the spike numbers, the average instantaneous frequency, and the average discharge frequency were markedly increased (P<0.01). There were no significant alterations in the membrane properties and intrinsic excitability induced by depolarized currents of small-sized neurons between groups. Compared with the control group, the whole-cell membrane currents, spike numbers, and the average instantaneous frequency were significantly increased in the model group(P<0.05, P<0.01) while rheobase was significantly decreased (P<0.05) in DRG medium-sized neurons labeled with biotin and CGRP. CONCLUSION: After the mice were modeled by ISO, the DRG medium-size neurons in the T1-T5 segment of the spinal cord may mediate the sensitization of acupoints on the body surface through their different neuronal membrane properties and intrinsic excitabilities.

[Involvement of sympathetic nerve activation in acupoint application induced improvement of gastric ulcer and motility in rats with gastric ulcer].

OBJECTIVE: To observe the effect of mustard oil application at "Liangmen" (ST21) on gastric ulcer (GU) and gastric motility and its association with the sympathetic nerve activity in rats with GU, so as to provide experimental basis for improvement of GU by acupoint application. METHODS: Thirty-nine male SD rats were randomly divided into control (n=7), model, acupoint application (AA), medication (guanethidine, an adrenergic sympathetic antagonist) and AA+medication groups (n=8 in each of the latter 4 groups). The GU model was made by applying acetic acid-immersed filter paper onto the gastric antrum. For rats of the AA and AA+medication groups, 50% mustard oil was applied to left ST21 for 10 min, once a day, for 9 consecutive days. Rats of the medication and AA+medication groups received intraperitoneal injection of guanethidine solution (40 mg/kg) beginning from the modeling day on, once a day for 10 consecutive days. The rat's body weight of each group was recorded on the 0th, 1st, 3rd, 7th and 9th day. The intragastric peristaltic wave frequency and the myoelectrical activity (frequency of slow waves, and integration of fast waves) of the gastric smooth muscle were recorded by using PowerLab data acquisition system. The gastric ulcer area was measured after the rats were executed, and histopathological changes of gastric antrum tissues (histopathological score including epithelial cell injury, submucosal edema, hemorrhagic injury, inflammatory cell infiltration score) were observed after H.E. staining. RESULTS: Compared with the control group, the body weight ratio, frequency of gastric peristaltic waves and slow wave frequency of gastric smooth muscle were significantly decreased (P<0.001, P<0.05), while the ulcer area, total histopathological score, epithelial cell injury score, submucosal edema score, hemorrhagic injury score and inflammatory cell infiltration score were significantly increased in the model group (P<0.05, P<0.001). Relevant to the model group, the AA group had a significant increase in the body weight ratio, frequency of gastric peristaltic waves, slow wave frequency, integration of fast waves (P<0.05, P<0.001), and a considerable decrease in the ulcer area, total histopathological score, epithelial cell injury score, submucosal edema score, hemorrhagic injury score and inflammatory cell infiltration score (P<0.05, P<0.001), and the medication group has a significantly decrease in the frequency of gastric peristaltic waves (P<0.05). In comparison with the AA group, the body weight ratio, frequency of gastric peristaltic waves and slow wave frequency of gastric smooth muscle in both medication and AA+medication groups, and the integration of fast waves in the medication group were obviously lower (P<0.05, P<0.001, P<0.01), whereas the levels of ulcer area, total pathological score, submucosal edema score, hemorrhagic injury score and inflammatory cell infiltration score in both medication and AA+medication groups, and the epithelial cell injury score in medication group were significantly higher (P<0.05, P<0.001). CONCLUSION: Application of mustard oil at acupoint ST21 can effectively remit GU caused by acetic acid and regulate gastric rhythmic contraction, which was mediated by sympathetic nerve.

Peripheral neural mechanism of visceral pain and acupoint sensitization in 2, 4, 6-trinitrobenzene sulfonic acid-induced colitis model mice. / ç»“è‚ ç‚Žæ¨¡åž‹å°é¼ å† è„ç—›åŠç©´ä½æ•åŒ–çš„å¤–å‘¨ç¥žç»æœºåˆ¶æŽ¢è®¨.

OBJECTIVES: To explore the neural mechanism of visceral pain and related somatic (acupoints) sensitization by using in vivo calcium imaging of dorsal root ganglia (DRG) neurons. METHODS: Eight BALB/c mice were randomly divided into control and model groups, with 4 mice in each group. The colitis model was induced by colorectal perfusion of 2, 4, 6-trinitrobenzene sulfonic acid (TNBS) once daily for 7 days. Mice of the control group received colorectal perfusion of normal saline once daily for 7 days. The location and area of the somatic neurogenic inflammation (cutaneous exudation of Evans blue ï¼»EBï¼½) of the 2 groups of mice were observed after intravenous injection of EB. For pain behavioral tests, sixteen C57BL/6J mice were randomly divided into control and model groups, with 8 mice in each group, and a Von Frey filament was used to stimulate the referred somatic reactive regions in colitis mice, and the number of avoidance and paw withdraw reaction within 10 tests was recorded. For in vivo DRG calcium imaging tests, 24 Pirt-GCaMP6s transgenic mice were randomly and equally divided into control group and colitis model group. The responses of the neurons in L6 or L4 DRG to colorectal distension (CRD), lower back brushing, or mechanical stimulation at the hindpaw were observed using confocal fluorescence microscope. RESULTS: Compared with the control group, the area of EB exudation spot in the hindpaw and lower back regions was increased in the colitis model group (P<0.05), and the avoidance or paw withdraw numbers induced by Von Frey stimulation at the lower back and hindpaw were increased (P<0.01, P<0.05), indicating that colitis induced regional skin (acupoints) sensitization in the lower back and hindpaw regions. Compared with the control group, the percentage of L6 DRG neurons activated by 60 mm Hg CRD in the colitis model mice were apparently increased (P<0.01), the activated neurons mainly involved the medium-sized DRG neurons (P<0.01). In Pirt-GCaMP6s transgenic mice, following brushing the skin of the receptive field (lower back) of L6 DRG neurons, the fluorescence intensity of the brushing-activated DRG neurons and small, medium and large-sized neurons were significantly higher in the colitis model group than those in the control group (P<0.001, P<0.01, P<0.05). After brushing and clamping the skin of the right hindpaw (receptive field of L4 DRG neurons), the percentages of the activated L4 DRG neurons were obviously higher in the colitis model group than those in the control group (P<0.01, P<0.05), while there were no significant changes in the proportion of small, medium and large-sized neurons between the control and colitis model groups. CONCLUSIONS: Colitis may lead to body surface sensitization at the same and adjacent neuro-segments as well as to an increase of the number and activity of the responsive lumbar DRG neurons, among which the L6 DRG neurons at the same neuro-segment as the rectum colon showed an increase in the number of responders and intensity of calcium fluorescence signal while L4 DRG neurons at the level adjacent to the rectum colon showed an increase in the number of responders, suggesting that there may be different mechanisms of peripheral neural sensitization.

Sino-European transcontinental basic and clinical high-tech acupuncture studies-part 3: violet laser stimulation in anesthetized rats.

The aim of this study was to determine the effect of violet laser stimulation on three acupuncture points in anesthetized rats and to test the hypothesis that violet laser light can modulate neurovegetative parameters like heart rate (HR), heart rate variability (HRV), and mean arterial blood pressure (MAP). Recordings were performed in 10 male anesthetized rats under three conditions in Beijing, and monitored with equipment from Graz, where also data analysis was performed. For stimulation a violet laser (emitted wavelength 405 nm, laser output 1 mW, continuous mode) was used. The electrocardiograms were recorded by an HRV Medilog AR12 system during laser acupuncture stimulation of the head, ear, and body (Baihui, "heart" ear acupoint, Zusanli). HR changed significantly only during (P = 0.013) and after (P = 0.038) stimulation at Baihui. Total HRV and the low frequency/high frequency ratio showed insignificant changes. There was an insignificant decrease in MAP after stimulation of Baihui acupoint. Violet laser stimulation offers a method to induce acute effects in HR and HRV in rats. Although the precise mechanism of this effect remains to be determined, alterations are significant. Violet laser stimulation on the Baihui acupoint could readily be translated to clinical studies.

Sino-European transcontinental basic and clinical high-tech acupuncture studies-part 1: auricular acupuncture increases heart rate variability in anesthetized rats.

Evidence-based research concerning the effects of high-tech acupuncture on autonomic function was performed by two research teams from China and Austria. This study describes the first transcontinental teleacupuncture measurements in animals. Heart rate (HR) and heart rate variability (HRV) recordings in 10 male Sprague-Dawley anesthetized rats were performed under stable conditions in Beijing, China, and the data analysis was completed in Graz, Austria. The electrocardiograms (ECGs) were recorded by an HRV Medilog AR12 system during acupuncture of the ear and body (PC6 Neiguan, CV12 Zhongwan, ST36 Zusanli). The data were analyzed using specially adapted novel Austrian software. HR did not change significantly during any acupuncture stimulation in anesthetized rats (ear acupuncture, PC6, CV12, or ST36). Total HRV only changed significantly (P = 0.025) during auricular acupuncture (acupoint heart). The low-frequency/high-frequency ratio parameter decreased significantly (P = 0.03) during stimulation of ST36. This change was based on intensification of the related mechanism of blood pressure regulation that has been demonstrated in previous studies in humans. Modernization of acupuncture research performed as a collaboration between China and Austria has also been demonstrated.

The effects of scraping therapy on local temperature and blood perfusion volume in healthy subjects.

Objective. We aim to study the therapeutic effects of scraping by investigating the changes of temperature and local blood perfusion volume in healthy subjects after scraping stimulation, and to explore the mechanism of scraping stimulation from the points of microcirculation and energy metabolism. Methods. Twenty-three health subjects were included in this study. Local blood perfusion volume and body surface temperature was detected at 5 min before scraping stimulation, 0, 15, 30, 60 and 90 min after scraping using Laser Doppler imager and infrared thermograph. Results. Significant increase was noted in the blood perfusion volume in the scraping area within 90 minutes compared to the baseline level and non-scraping area (P < 0.001). Compared with non-scraping area, an increase of body temperature with an average of 1°C was observed after scraping stimulation (P < 0.01). Conclusion. Scraping can significantly improve the blood perfusion volume and increase the temperature in the scraping area, promoting the local blood circulation and energy metabolism.

Brain-modulated effects of auricular acupressure on the regulation of autonomic function in healthy volunteers.

Auricular acupuncture has been described in ancient China as well as Egypt, Greece, and Rome. At the end of the 1950s, ear acupuncture was further developed by the French physician Dr. Paul Nogier. The goal of this study was to develop a new system for ear acupressure (vibration stimulation) and to perform pilot investigations on the possible acute effects of vibration and manual ear acupressure on heart rate (HR), heart rate variability (HRV), pulse wave velocity (PWV), and the augmentation index (AIx) using new noninvasive recording methods. Investigations were performed in 14 healthy volunteers (mean age ± SD: 26.3 ± 4.3 years; 9 females, 5 males) before, during, and after acupressure vibration and manual acupressure stimulation at the "heart" auricular acupuncture point. The results showed a significant decrease in HR (P ≤ 0.001) and a significant increase in HRV total (P = 0.008) after manual ear acupressure. The PWV decreased markedly (yet insignificantly) whereas the AIx increased immediately after both methods of stimulation. The increase in the low-frequency band of HRV was mainly based on the intensification of the related mechanism of blood pressure regulation (10-s-rhythm). Further studies in Beijing using animal models and investigations in Graz using human subjects are already in progress.

[Role of animal experiment in acupuncture translational medicine].

The basic biological research of acupuncture originates from clinical practice and serves clinical application. It is the fundamental idea of "acupuncture translational medicine". The animal experiment is indispensable in acupuncture translational medicine, which contributes to simplifying the complex factors affecting the clinical efficacy of acupuncture, observing the effect of the nociceptive stimuli and obtaining the invasive and off-body biomarkers. Focusing on animal experiment, the basic biological research can objectively reveal the structure and function of acupoints, obtain the parameters required by different stimulation methods and clarify the material basis which acupuncture works, so as to provide the references and evidences for acupoint combination and the selection of appropriate stimulation in clinical practice, as well as the development of medical instruments and new drugs.

[Effect of electroacupuncture on mood and estrogen receptor α expression in mice with polycystic ovary syndrome induced by bisphenol A exposure].

OBJECTIVE: To compare the effect of electroacupuncture (EA) on "Guanyuan" (CV4) or sensitization points in mice with polycystic ovary syndrome (PCOS), so as to explore its mechanisms underlying improvement of PCOS. METHODS: In the first part of this study, 26 female ICR mice were randomized into control group (8 mice) and model group (18 mice). The PCOS model was established by gavage of bisphenol A (BPA) at a dose of 100 mg/kg, and the control group were gavage of equal volume of corn oil, once daily, 5 days a week for 4 conse-cutive weeks. Evans blue (EB) dye (0.1 mL/10 g) was injected into the caudal vein after modeling. The size, number and distribution of EB exudation points at the skin were observed. In the second part of this study, 32 mice were randomized into control, model, EA-CV4 and EA-sensitization points groups (8 mice in each group). EA (2 Hz, 2 mA) was applied to the sensitization points or CV4 for 20 min, once daily, 5 days a week for 4 conse-cutive weeks. The body weight was measured once a week for 8 consecutive weeks. The behavior changes were evaluated by open field test and elevated plus maze test. H.E. staining was used to observe the histopathologic changes of the ovary tissue. Serum level of estradiol (E2) was measured by ELISA. The expressions of estrogen receptor α (ER-α) in ovarian and uterine tissues were detected by Western blot and immunofluorescence. RESULTS: (1) In PCOS mice, the EB exudation points were found to overlap the lower abdomen, lumbosacral, chest, back and lower limbs regions, and the number of EB points was significantly more than that of the control group (P<0.01). (2) After the intervention and compared with the control group, the ovaries showed polycystic changes and an increase of atresia follicles with a larger diameter, the activity time in the central area, the total distance of movement, the times of open-arm entries, the duration in open-arm, the serum E2 content and the expression of ER-α in ovarian tissue were significantly decreased (P<0.01, P<0.05), while the mice's body weight and the expression of ER-α in uterine tissue were increased (P<0.05) in the model group. After the intervention and compared with the model group, a small number of normal follicles and corpus luteum were observed under microscope, the activity time in the central area, the total distance of movement, the times of open-arm entries, the duration in open-arm, the serum E2 content and the expression of ER-α in ovarian tissue were significantly increased (P<0.05, P<0.01), while the mice's body weight and the expression of ER-α in uterine tissue was decreased (P<0.05) in both EA-CV4 and EA-sensitization points groups. CONCLUSION: EA at sensitization points and CV4 can regulate the expression of estrogen and ER-α in PCOS mice, and improve the anxiety like behavior. EB exudation points on the body surface can not only reflect the functional changes of organs, but also treat diseases through body surface stimulation, suggesting a dual role in diagnosis and treatment.

[Comparison of effects of routine electroacupuncture and pre-electroacupuncture in improving lung function in acute lung injury rats].

OBJECTIVE: To observe the effect of electroacupuncture (EA) and pre-EA at"Taiyuan"(LU9) on histopathological changes, pulmonary ventilation function, pneodynamics, and contents of TNF-α, IL-1ß and IL-10 in the bronchoalveolar lavage fluid in rats with acute lung injury (ALI), in order to explore their differences of therapeutic effects. METHODS: A total of 92 male SD rats were randomly divided into control, model, routine EA and pre-EA groups, with 23 rats in each group. The ALI rat model was established by instillation of 0.5% lipopolysaccharide solution (5 mg/kg) intratracheally. EA (3 mA, 10 Hz) was applied to bilateral LU9 for 20 min, once 5 h after modeling for rats of the routine EA group, and once on the 1st, 3rd and 5th day respectively before modeling for rats of the pre-EA group. The pulmonary ventilation functions, including dynamic lung compliance (Cdyn), tidal volume (TV)，minute ventilation volume (MV), lung resistance, peak inspiratory flow (PIF), peak expiratory flow (PEF), were measured by using a small animal lung function detection system. The electromyogram (EMG) of the diaphragm muscle was recorded for observing changes of the total integral electromyogram(iEMG), single iEMG and respiration rate. Pathological changes of the lung tissue were scored after H.E. staining. The contents of TNF-α, IL-1ß and IL-10 in the bronchoalveolar lavage fluid were determined by enzyme linked immunosorbent assay. RESULTS: Compared with the control group, the Cdyn, TV, MV and PIF consi-derably decreased (P<0.000 1), and the lung resistance, total iEMG of diaphragm muscle, lung injury score, and contents of TNF-α and IL-1ß were significantly increased (P<0.05, P<0.001, P<0.01) in the model group. In comparison with the model group, the TV, MV and PIF in both routine EA and pre-EA groups,and the Cdyn, total iEMG and single iEMG in the routine EA group, as well as the IL-10 content in the pre-EA group were significantly increased (P<0.01, P<0.001, P<0.05), whereas the lung injury score, and contents of TNF-α and IL-1ß were significantly decreased in the pre-EA group (P<0.05). CONCLUSION: Both pre-EA and routine EA at LU9 can improve the lung function of ALI model rats. Pre-electroacupuncture is related to local immunoregulation, while electroacupuncture is mainly related to the improvement of pulmonary ventilation function and respiratory motility.

[Association between skin microecology and acupoint sensitization].

The skin microecology interacts with the immune system for a lifetime, and the skin microecology is in dynamic balance in the physiological state. The flora with the same physical and chemical properties on the surface of the skin can maintain a relatively stable state in a certain period of time, and there are dynamic changes of microflora in both physiology and disease state. Combined with the study of the relationship among skin microorganisms, skin diseases and visceral diseases, the relationship between skin microecological changes and visceral pathological state is explored. On this basis, it is proposed that the sensitized acupoint is an abnormal manifestation of the body homeostasis imbalance on the body surface, and the local substance variation on the sensitization acupoint may induce microbial changes. In terms of treatment, moxibustion and other surface treatments that function through immunity may also involve microorganisms.

[Roles of nociceptors in acupoint sensitization: recent advances].

Hyperalgesia and functional plasticity are the important components of acupoint sensitization. Reveal of the neuromechanism of acupoint sensitization may play a positive role in promoting the development of acupuncturology in the world. The nociceptors, including Aδ and C subtypes distributing in the acupoint region and target organs, are responsible for the transmission of signals of peripheral noxious stimuli and acupuncture-liking stimulation to the dorsal horns of the spinal cord and supraspinal levels. A previous study reveals that the C type nociceptors are involved in the acupoint sensitization. Recent studies indicate that there exists a subtype of mechanical responsiveness in the C type receptors, named "silent nociceptor" which is awa-kened when diseases occur, being very similar to the dynamic sensitization characteristics of acupoints. Hence, we, in the present review, make a discussion about the role of C-type silent nociceptor in the hyperalgesia and functional plasticity of the sensitized acupoint according to previous studies and recent advances, so as to provide more ideas and opportunities for the investigation on the scientific characteristics of acupoints.

Increasing 28 mitogenomes of Ephemeroptera, Odonata and Plecoptera support the Chiastomyaria hypothesis with three different outgroup combinations.

BACKGROUND: The phylogenetic relationships of Odonata (dragonflies and damselflies) and Ephemeroptera (mayflies) remain unresolved. Different researchers have supported one of three hypotheses (Palaeoptera, Chiastomyaria or Metapterygota) based on data from different morphological characters and molecular markers, sometimes even re-assessing the same transcriptomes or mitochondrial genomes. The appropriate choice of outgroups and more taxon sampling is thought to eliminate artificial phylogenetic relationships and obtain an accurate phylogeny. Hence, in the current study, we sequenced 28 mt genomes from Ephemeroptera, Odonata and Plecoptera to further investigate phylogenetic relationships, the probability of each of the three hypotheses, and to examine mt gene arrangements in these species. We selected three different combinations of outgroups to analyze how outgroup choice affected the phylogenetic relationships of Odonata and Ephemeroptera. METHODS: Mitochondrial genomes from 28 species of mayflies, dragonflies, damselflies and stoneflies were sequenced. We used Bayesian inference (BI) and Maximum likelihood (ML) analyses for each dataset to reconstruct an accurate phylogeny of these winged insect orders. The effect of outgroup choice was assessed by separate analyses using three outgroups combinations: (a) four bristletails and three silverfish as outgroups, (b) five bristletails and three silverfish as outgroups, or (c) five diplurans as outgroups. RESULTS: Among these sequenced mitogenomes we found the gene arrangement IMQM in Heptageniidae (Ephemeroptera), and an inverted and translocated tRNA-Ile between the 12S RNA gene and the control region in Ephemerellidae (Ephemeroptera). The IMQM gene arrangement in Heptageniidae (Ephemeroptera) can be explained via the tandem-duplication and random loss model, and the transposition and inversion of tRNA-Ile genes in Ephemerellidae can be explained through the recombination and tandem duplication-random loss (TDRL) model. Our phylogenetic analysis strongly supported the Chiastomyaria hypothesis in three different outgroup combinations in BI analyses. The results also show that suitable outgroups are very important to determining phylogenetic relationships in the rapid evolution of insects especially among Ephemeroptera and Odonata. The mt genome is a suitable marker to investigate the phylogeny of inter-order and inter-family relationships of insects but outgroup choice is very important for deriving these relationships among winged insects. Hence, we must carefully choose the correct outgroup in order to discuss the relationships of Ephemeroptera and Odonata.

[Acupoint sensitization enhances inhibitory effect of electroacupuncture on jejunum mobility in rats].

OBJECTIVE: To observe the effect difference of electroacupuncture (EA) at the sensitized and non-sensitized acupoint "Tianshu" (ST25) on the motility of jejunum in rats. METHODS: Male SD rats were randomly divided into blank control, sensitized ST25 (sensitization), and non-sensitized ST25 (non-sensitization) groups (n=16 in each group). The sensitization and non-sensitization were induced by injection of 15% mustard oil (20 µL) and Paraffin oil (20 µL) into the left ST25 area respectively. The rats' behavior reactions were assessed by recording the numbers and duration of licking the local sensitized skin region. At the end of experiments and after regular trans-cardiac perfusion and fixing with 10% neutral formaldehyde, the skin and muscular tissues of the ST25 region (1 cm×1 cm) were taken for H.E. staining to observe the local histopathologic changes. The intestinal motility was detected by recording the contraction pressure waves of jejunum through a small balloon-connected pressure transducer and an amplifier, followed by calculating the ratios of amplitude and frequency of contraction waves between pre- and post-EA stimulation. EA (2 Hz, 0.2-6.0 mA) was applied to the sensitized and non-sensitized ST25 regions for 20 s for comparing their effects on jejunum motility. RESULTS: Compared with the blank control group, mustard oil injection but not Paraffin oil injection resulted in histological edema and neutrophils infiltration in tissues of ST25 region, and also striking increase of the number and duration of licking (P<0.01), suggesting a sensitization of ST25 area after mustard oil injection. The ratios of spontaneous contraction amplitude and frequency of jejunum were remarkably decreased in mustard oil-treated rats compared to baseline and Paraffin oil group (P<0.001，P<0.01). The maximum inhibitory effect of EA on the intestinal movement amplitude was significantly higher in the sensitization group than in the blank control and non-sensitization groups (P<0.05，P<0.01), and there was a dose-effect relationship between the current intensity of EA (0.5-3.0 mA) and the inhibitory rate 50% of the contraction amplitude (not the frequency) in the sensitization group (P<0.001), but not in the blank control and non-sensitization groups (P>0.05). Additionally, 1.5 mA EA stimulation at the sensitized ST25 (not at the non-sensitized ST25) had an inhibitory effect on the contraction amplitude (P<0.05) rather than on the contraction frequency (P>0.05) in comparison with the blank control group. CONCLUSION: Sensitization of acupoint ST25 can modulate the motility of jejunum and enhances the inhibitory effect of EA on the contraction amplitude of jejunum in rats. The inhibitory effect of sensitized ST25 EA is evidently stronger than that of the non-sensitized ST25 EA.

Effects of different acupuncture stimulations on NO content in acupoint areas.

OBJECTIVE: To study the effects of different acupuncture stimulation on nitric oxide (NO) content in acupoint areas. METHODS: The experiment was performed in 47 healthy volunteers providing informed consents. Both males and females were included in each group: 11 cases in electro-acupuncture group, 13 cases in manual-acupuncture group, 16 cases in warm-moxibustion group, and 7 cases in control group. Stimulations were given at Jianshi (PC 5) of the Pericardium Meridian and Chengshan (BL 57) of the Bladder Meridian with manual-acupuncture, electro-acupuncture and warm-moxibustion respectively. NO was collected from Ximen (PC 4) and Chengjin (BL 56) areas on the ipsilateral meridians, and the content was detected. RESULTS: The content of NO in the warm-moxibustion group was higher than that in the electro-acupuncture group, the content in electro-acupuncture group was higher than that in the manual acupuncture group (P < 0.05). CONCLUSION: All the three stimulations can raise NO levels on acupoint areas significantly, and the effect moxibustion is superior to that of electro-acupuncture and manual acupuncture.

[Electroacupuncture at "Neiguan"(PC6) decreased cardiac sympathetic hyperactivity and improved cardiac function in chronic myocardial ischemia model rats].

OBJECTIVE: To investigate the autonomic nervous mechanism of acupuncture therapy in the treatment of ischemic cardiomyopathy by observing the influence of electroacupuncture (EA) at "Neiguan"(PC6) on the superior cervical cardiac nerve activity and cardiac function in chronic myocardial ischemia (CMI) rats. METHODS: Male SD rats were randomly divided into control, model and EA groups (n＝8 in each group). The CMI model was established by ligating the anterior descending branch of the left coronary artery. EA (15 Hz, 1.5 mA) was applied to bilateral PC6 for 20 min, once a day for 28 consecutive days. Cardiac sympathetic nerve electrical activities (CSNEA), electrocardiogram (ECG) of the standard limb lead Ⅱ and ultrasonic cardiogram (UCG) were recorded for observing changes of ST segment height, left ventricular ejection fraction (LVEF) and left ventricular fractional shortening (LVFS)．. RESULTS: The CSNEA was significantly increased(P<0.001), and the hight of ECG-STⅡ， LVEF and LVFS were considerably decreased in the model group relevant to the control group (P<0.01, P<0.001), while after the intervention, modeling induced increase of CSNEA and decrease of ECG-STⅡ， LVEF and LVFS were obviously suppressed in the EA group in comparison with the model group (P<0.01, P<0.05, P<0.001). CONCLUSION: EA of PC6 can improve cardiac function and myocardial ischemia in CMI rats, which is possibly related to its effect in decreasing cardiac sympathetic hyperactivity.