[Effects of moxa smoke through olfactory pathway on learning and memory ability in rapid aging mice].

OBJECTIVE: To observe the effects of moxa smoke through olfactory pathway on learning and memory ability in rapid aging (SAMP8) mice, and to explore the action pathway of moxa smoke. METHODS: Forty-eight six-month-old male SAMP8 mice were randomly divided into a model group, an olfactory dysfunction group, a moxa smoke group and an olfactory dysfunction + moxa smoke group, with 12 mice in each group. Twelve age-matched male SAMR1 mice were used as the blank group. The olfactory dysfunction model was induced in the olfactory dysfunction group and the olfactory dysfunction + moxa smoke group by intraperitoneal injection of 3-methylindole (3-MI) with 300 mg/kg, and the moxa smoke group and the olfactory dysfunction + moxa smoke group were intervened with moxa smoke at a concentration of 10-15 mg/m3 for 30 min per day, with a total of 6 interventions per week. After 6 weeks, the emotion and cognitive function of mice was tested by open field test and Morris water maze test, and the neuronal morphology in the CAI area of the hippocampus was observed by HE staining. The contents of neurotransmitters (glutamic acid [Glu], gamma-aminobutyric acid [GABA], dopamine [DA], and 5-hydroxytryptamine [5-HT]) in hippocampal tissue of mice were detected by ELISA. RESULTS: The mice in the blank group, the model group and the moxa smoke group could find the buried food pellets within 300 s, while the mice in the olfactory dysfunction group and the olfactory dysfunction + moxa smoke group took more than 300 s to find them. Compared with the blank group, the model group had increased vertical and horizontal movements (P<0.05) and reduced central area residence time (P<0.05) in the open field test, prolonged mean escape latency on days 1-4 (P<0.05), and decreased search time, swimming distance and swimming distance ratio in the target quadrant of the Morris water maze test, and decreased GABA, DA and 5-HT contents (P<0.05, P<0.01) and increased Glu content (P<0.05) in hippocampal tissue. Compared with the model group, the olfactory dysfunction group had increased vertical movements (P<0.05), reduced central area residence time (P<0.05), and increased DA content in hippocampal tissue (P<0.05); the olfactory dysfunction + moxa smoke group had shortened mean escape latency on days 3 and 4 of the Morris water maze test (P<0.05) and increased DA content in hippocampal tissue (P<0.05); the moxa smoke group had prolonged search time in the target quadrant (P<0.05) and increased swimming distance ratio, and increased DA and 5-HT contents in hippocampal tissue (P<0.05, P<0.01) and decreased Glu content in hippocampal tissue (P<0.05). Compared with the olfactory dysfunction group, the olfactory dysfunction + moxa smoke group showed a shortened mean escape latency on day 4 of the Morris water maze test (P<0.05). Compared with the moxa smoke group, the olfactory dysfunction + moxa smoke group had a decreased 5-HT content in the hippocampus (P<0.05). Compared with the blank group, the model group showed a reduced number of neurons in the CA1 area of the hippocampus with a disordered arrangement; the olfactory dysfunction group had similar neuronal morphology in the CA1 area of the hippocampus to the model group. Compared with the model group, the moxa smoke group had an increased number of neurons in the CA1 area of the hippocampus that were more densely packed. Compared with the moxa smoke group, the olfactory dysfunction + moxa smoke group had a reduced number of neurons in the CA1 area of the hippocampus, with the extent between that of the moxa smoke group and the olfactory dysfunction group. CONCLUSION: The moxa smoke could regulate the contents of neurotransmitters Glu, DA and 5-HT in hippocampal tissue through olfactory pathway to improve the learning and memory ability of SAMP8 mice, and the olfactory is not the only effective pathway.

Effects of electroacupuncture on bladder dysfunction and the expression of PACAP38 in a diabetic rat model.

Objective: To explore the effects and the possible mechanism of electroacupuncture (EA) on diabetic bladder dysfunction (DBD) in streptozotocin-high fat diet (STZ-HFD) induced type 2 diabetes mellitus (T2DM) rats. Methods: The experiment was divided into Control, diabetic bladder dysfunction, electroacupuncture, and Sham electroacupuncture group. After 8 weeks of electroacupuncture intervention, the body mass, 24 h urine volume, intraperitoneal glucose tolerance test (IPGTT), and urodynamics were detected. After the wet weight of the bladder was detected, the hematoxylin-eosin (HE), Masson's trichrome, and TUNEL were used to analyze histological changes. The PACAP38 expressions in the bladder were detected by Real-time PCR and Western blot. Results: Compared to the Control group, the bladder wet weight, 24 h urine volume, blood glucose, maximum bladder capacity, bladder compliance, bladder wall thickness, the smooth muscle/collagen ratio, and apoptosis rate of the diabetic bladder dysfunction group were significantly increased. Moreover, the body mass and leak point pressure were significantly reduced. Compared with the Sham electroacupuncture group, the bladder wet weight, maximum bladder capacity, bladder compliance, bladder wall thickness, and apoptosis rate of the electroacupuncture group were significantly reduced. In contrast, the leak point pressure was increased. The PACAP38 mRNA and PACAP38 protein expression of the diabetic bladder dysfunction group were significantly lower than the Control group, while electroacupuncture treatment could upregulate PACAP38 mRNA levels and PACAP38 protein expression of diabetic bladder dysfunction model rats. Conclusion: electroacupuncture could ameliorate bladder dysfunction in the diabetic bladder dysfunction model rats by reversing bladder remodeling, which might be mainly mediated by regulating the PACAP38 level.

Effect of Moxibustion on Behavioral Changes and Expression of APP and BACE1 in Hippocampus of SAMP8 Mice.

OBJECTIVE: To observe the effect of moxibustion on cognitive function of aging mice, to observe the effect of moxibustion on protein and gene expression of APP metabolism pathway, and to explore the mechanism of action in moxibustion. METHODS: 24 SAMP8 were randomly divided into 2 groups (12 in each group): moxibustion group and model group. 12 SAMR1 mice were used as blank controls. Mice in the moxibustion group were treated with moxibustion for 8 weeks, 10 minutes each time, 5 times a week, and for a total of 8 weeks. The model group and the blank group were treated with sham-moxibustion. Behavior tests were used to detect the learning and memory ability of each group of mice. Immunohistochemical, western blot, and RT-PCR were used to detect the protein and mRNA expression of APP and BACE1. Furthermore, the expressions of miR-29 and miR-101 were observed by RT-PCR method to explore the mechanism of moxibustion at the genetic level. RESULTS: In this study, relative to normal mice, we found that aging mice showed behavioral changes consistent with the onset of AD. However, moxibustion interventions were able to mitigate these effects to some degree in aging mice. In addition, moxibustion was proved to regulate APP metabolism pathway at protein and gene level through molecular biology tests. CONCLUSION: The data suggest that the effect of moxibustion intervention on cognitive function in aging mice is related to the regulation of genes and proteins involved in APP metabolism pathway; this may be a potential target for treating Alzheimer's disease.

Effect of Moxibustion on Hyperhomocysteinemia and Oxidative Stress Induced by High-Methionine Diet.

OBJECTIVE: The aim of this study was to assess the effects of moxibustion on the animal model of oxidative stress and cardiovascular injury induced by high-methionine diet (2% methionine and 3.5% fat on the basis of ordinary maintenance feed) during 12 weeks. METHODS: 53 mice were divided into four groups: mice in the Control group (n = 8), mice in the Met group (n = 8), mice in the Met group (n = 8), mice in the Met group (n = 8), mice in the Met group (. RESULTS: Compared with the Met group, our results indicated that through moxibustion intervention, the content of serum Hcy and its intermediate metabolite SAH can be reduced to a certain extent, and SOD, HO-1, and ox-LDL can be increased. CONCLUSION: This study showed moxibustion's ability to enhance the body's antioxidation and protect vascular endothelial function, thus playing an early role in the prevention and treatment of atherosclerosis.

Moxibustion at 'Danzhong' (RN17) and 'Guanyuan' (RN4) for fatigue symptom in patients with depression: Study protocol clinical trial (SPIRIT Compliant).

BACKGROUND: Fatigue is one of the most prevalent and debilitating symptoms of major depressive disorder (MDD). The effective management of depression-related fatigue has an important impact on the patient's abilities, functioning, and quality of life (QOL). Moxibustion has been widely used in Traditional Chinese Medicine to manage fatigue. Recent studies have also demonstrated that moxibustion is effective for treating cancer-related fatigue and chronic fatigue syndrome. However, there is not sufficient data supporting the effect of moxibustion for depression-related fatigue. Therefore, this randomized, assessor-blinded, wait-list controlled trial is designed to evaluate the effectiveness, safety, and feasibility of moxibustion treatment for depression-related fatigue. METHODS: One hundred and seventy-six participants who meet the diagnostic criteria for depression in the International Classification of Diseases, tenth revision (ICD-10), and who also have a score of ≥1 on the 13 item of the Hamilton Depression Rating Scale-17 (HAMD-17), will be enrolled. At study entry, participants will undergo anti-depressant treatment for at least 1 month. Then those who still have a score of ≥1 on the 13 item of the HAMD-17 will be randomly allocated to either a moxibustion group or wait-list control group in a ratio of 1:1. Anti-depressants will be provided for both groups during the whole process of the study period. Participants in the moxibustion group will undergo 14 sessions of moxibustion (over 2 weeks) with anti-depressant treatment, and participants in the wait-list control group will receive only anti-depressant treatment. Subsequently, participants in the moxibustion group will be followed-up for 4 weeks. The primary outcome measure will be the Fatigue Severity Scale (FSS). The secondary outcome measure will be the HAMD-17. Safety will be assessed by monitoring adverse events during the study. Trial feasibility will also be assessed in this study. DISCUSSION: The results of this study may provide evidence for the efficacy of moxibustion as an adjunct to antidepressants for depression-related fatigue, and promote a more widespread foundation for the selection of moxibustion in the clinical setting as well as for future research in moxibustion therapy. TRIAL REGISTRATION: This study protocol was registered at the Chinese Clinical Trial Registry (ChiCTR1800016905).

Moxibustion therapy for treating patients with primary osteoporosis: A systematic review and meta-analysis protocol.

BACKGROUND: Primary osteoporosis (POP) is a common disease among elderly, which increase the risk of fracture and impact to the quality of life. As a Chinese traditional therapy, moxibustion has been commonly applied in treating chronic musculoskeletal diseases in China. Many trails have shown that moxibustion therapy is effective in treating primary osteoporosis. The protocol aims to present the methods used to access the effectiveness and safety of moxibustion therapy for patients with primary osteoporosis. METHODS: The following databases will be searched from their inception: the Cochrane Central Register of Controlled Trails(CENTRAL), Pubmed, EMBASE, China National Knowledge Infrastructure(CNKI), Chinese Biomedical Literature Database(CBM), Chinese Scientific Journal Database (VIP database), and Wan-Fang Database. Clinical randomized controlled trials related to moxibustion therapy for treating primary osteoporosis will be included, regardless of publication status and languages. Study selection, data collection, and quality assessment will be independently conducted by 2 researchers. We will select the fixed-effects or random-effects model according to the heterogeneity assessment for data synthesis. Bone mineral density(BMD) will be the primary outcomes. Visual analogue scale(VAS), response rate, TCM Syndrome scale(TCMSS), bone gla protein(BGP), alkaline phosphatase(BALP), blood calcium(Ca), blood phosphate(P), quality of life(QOL) will be the second outcomes. If it is appropriate for meta-analysis, RevMan V.5.3 statistical software will be used. Otherwise, a systematic narrative synthesis will be conducted. The results will be presented as risk ratio (RR) with 95% confidence intervals (CIs) for dichotomous data and weight mean difference(WMD) or standard mean difference (SMD) 95% CIs for continuous data. TRIAL REGISTRATION NUMBER: PROSPERO CRD42019129507.

The efficacy and safety of moxibustion for primary dysmenorrhea: A systematic review protocol.

BACKGROUND: Primary dysmenorrhea (PDM) is one of the most prevalent gynecological diseases in women of childbearing age. The major medications treating PDM usually make sense and side effects, while moxibustion is known as a safe and effective treatment for PDM. This review aims to systematically evaluate the effect and safety of moxibustion for treating PDM. METHODS: We will search all randomized controlled trials for moxibustion therapy on PDM, electronically and manually, regardless of publication status, till October 31, 2019. Online databases include the Cochrane Central Register of Controlled Trials; PubMed; EMBASE; China National Knowledge Infrastructure; Chinese Biomedical Literature Database; Chinese Scientific Journal Database (VIP database); and Wan-Fang Database. Two reviewers will search these databases, select data and measure the quality of studies independently. The methodological quality will be assessed by the Cochrane Reviewer's Handbook 6.0. The primary outcomes include clinical efficacy and visual analog scale, and the secondary outcomes include adverse events and quality of life. Four reviewers will independently extract the data and assess the qualities of the studies. Statistical analysis will be conducted with R package for each outcome. Study selection, data extraction, and assessment of risk of bias will be performed independently by 2 reviewers. RESULTS: This study will provide a comprehensive review of the available evidence for the treatment of moxibustion with PDM. CONCLUSION: The conclusion of our study will provide updated evidence to judge whether moxibustion is an effective and safe intervention for patients with PDM. PROSPERO REGISTRATION NUMBER: CRD42019129993.

Effects of Moxibustion and Moxa Smoke on Behavior Changes and Energy Metabolism in APP/PS1 Mice.

OBJECTIVE: To investigate the antiaging effects of moxibustion and moxa smoke on APP/PS1 mice and to illustrate the mechanism of moxibustion improving Alzheimer's disease (AD). METHODS: 36 male APP/PS1 mice were randomly assigned into three groups (n = 12), including a model control group, a moxibustion group, and a moxa smoke group. In addition, 12 C57BL/6 normal mice served as a normal (negative) control group. Mice in the moxibustion group received moxibustion intervention using Guanyuan (RN4) acupoint. Mice in the moxa smoke group received moxa smoke exposure with the same frequency as the moxibustion group. Behavioral tests were implemented in the 9th week, 3 days after the completion of the intervention. Tricarboxylic acid cycle and fatty acid metabolomics assessments of the mice were determined after behavioral tests. RESULTS: In this study, relative to normal mice, we found that AD mice showed altered tricarboxylic and fatty acid metabolism and showed behavioral changes consistent with the onset of AD. However, both the moxibustion and moxa smoke interventions were able to mitigate these effects to some degree in AD mice. CONCLUSIONS: The data suggest that tricarboxylic acid cycle and unsaturated fatty acid metabolomics changes may be a target of AD, and the beneficial effects of moxibustion on cognitive behaviors may be mediated by the energy metabolism system.

Lung Function Decline after 24 Weeks of Moxa Smoke Exposure in Rats.

OBJECTIVE: Moxibustion is a complementary therapy that has been used for thousands of years. Burning moxa produces smoke and inhalable particulates. Recent research has indicated that smoke inhalation is associated with negative lung effects. This study aimed to evaluate the lung function of rats after moxa smoke exposure at different concentrations. METHODS: Using a randomised block experiment design, 28 male Wistar rats were randomly divided into three moxa smoke groups (opacity) (n=7): low concentration (27.45 mg/m3), medium concentration (168.76 mg/m3), and high concentration (384.67 mg/m3) with a control group. Rats in the moxa smoke groups were exposed in an automatic dynamic exposure device separately with different concentrations for 20 min/d, 6d/week, for 24 weeks. Rats in the control group were exposed in the same space without moxa smoke. Lung function was evaluated by the AniRes 2005 animal pulmonary function analysing system. Statistical Product and Service Solutions 18.0 software was used for data analysis. RESULTS: In the study, no deaths were found in any group. There was no difference of forced expiratory volume in one second/forced vital capacity percentage (FEV1/FVC%), inspiratory resistance (Ri), and expiratory resistance (Re) among each group after 24 weeks of moxa smoke exposure (P>0.05). Compared with the control group (0.33 ml/cmH20), dynamic compliance (Cdyn) was reduced in the medium (0.29 ml/cmH20) and high (0.25 ml/cmH20) concentration groups (P<0.05); however, Cdyn in the low concentration group (0.29 ml/cmH20) was not significantly affected. CONCLUSION: Moxa smoke exposure at low concentrations did not affect the rat's lung function. Moxa smoke of medium and high concentrations destroyed the lung function represented by decreased Cdyn. However, moxa smoke of low concentrations (27.45 mg/m3) is much higher than the concentration in a regular moxibustion clinic (3.54 mg/m3). Moxa smoke at higher concentrations might destroy the lung function. The safety evaluation of moxa smoke requires further research.

[Micronucleus in vitro induced by inhalable particulate matters in moxa smoke].

OBJECTIVE: To investigate whether inhalable particulate matters can cause the damage of chromosome or mitotic apparatus to produce micronucleus, and to evaluate genetic toxicology of moxa smoke on chromosome. METHODS: By MTT method, the 24 h half maximal inhibitory concentration (IC50) of moxa smoke condensation (MSC) on Chinese hamster ovary (CHO) cells was 0.087 mg/mL. CHO cells, which were cultured in vitro, were divided into a solvent control group, a positive control group (cyclophosphamide as solvent), a low concentration group, a moderate concentration group and a high concentration group. The low concentration group, moderate concentration group and high concentration group were set approximately 1/8, 1/4, 1/2 of IC50, respectively. Whether micronucleus had dose-effect response induced by the damage of chromosome or mitotic apparatus was observed after CHO cells were contaminated by MSC in the low concentration group, moderate concentration group and high concentration group. RESULTS: The rate of micronucleus induced by MSC in the low concentration group, moderate concentration group and high concentration group was higher than that in the solvent control group (all P < 0.05), which presented dosage-effect response. The experiment was repeated 3 times, indicating it was repeatable with statistical significance. CONCLUSION: High concentration of MSC shows toxicity to induce chromosome damage, which disappears at low concentration. The genetic toxicology is also dependent on concentration, and the concentration of moxa smoke is essential. In clinical treatment, it is noted to control the level of moxa smoke, while the clinical safety standard of moxa smoke concentration is in need of further study.

Effect of moxa smoke produced during combustion of Aiye (Folium Artemisiae Argyi) on behavioral changes in mice inhaling the smoke.

OBJECTIVE: To evaluate the effects that inhalation of the combustion products of the traditional Chinese herb Aiye (Folium Artemisiae Argyi) has on the central nervous system. METHODS: Forty Kunming mice (half male) were randomly assigned (n = 10/group) to a control group (C) or one of three moxa smoke concentration groups (% opacity): low (L1; 0.4% ), medium (M1; 2% ), and high (H1; 15% ). Mice in the latter three groups were exposed to moxa smoke in a dynamic gas exposure cabinet for 20 min per day for 7 days. Mice in control group were placed in the same cabinet without any intervention. For the sleep experiments, another 50 mice were divided into five groups of 10 mice each: a saline-injected control group, L1 + pentobarbital sodium (PS)-injected group, M1 + PS group, H1 (15% ) + PS group, and a positive control group (10 mg/kg, chlorpromazine, p.o.). The weight, general activities, locomotor activities, rotarod performance, sleep duration, and sleeping rate induced by a subthreshold dose of pentobarbital sodium were recorded in the mice, and the composition of moxa smoke was analyzed using headspace gas chromatography (GC-HS). RESULTS: A low concentration of smoke significantly decreased the frequency of locomotor activities and the time for which the mice remaining on the rotarod; however, a high smoke concentration significantly prolonged the pentobarbital-induced sleeping time and sleeping rate. CONCLUSION: The concentration-dependent relaxing effects of moxa smoke on the Central Nervous System (CNS) were confirmed. Moreover, GC-HS analysis showed that the component present in the highest concentration in moxa smoke was eucalyptol, an essential oil well recognized for its soothing effects on the CNS. This may therefore be accountable for the sedative effects of moxa smoke.

[Effects of moxa smoke with different concentrations on expression of SOD and MDA in lung and serum of rats].

OBJECTIVE: To observe the effects of intervention of moxa smoke with different concentrations on superoxide dismutase (SOD) and malondialdehyde (MDA) in serum and lung of male rats, so as to explore the safety concentration of moxa smoke. METHODS: A total of 32 Wistar male rats were randomly divided into a control group, a low-concentration group, a moderate-concentration group and a high-concentration group, 8 rats in each one. All the rats were exposed in the full-automatic toxicant exposure cabinet, and the overshadow of moxa smoke was set at 0%, 10%, 40% and 70%, respectively. Each rat was exposed for 20 min per day. After 26 weeks, the activities of SOD and content of MDA in serum, lung organ and bronchoalveolar lavage fluid were tested. RESULTS: Compared with the control group, the activities of serum SOD in the high-concentration group were reduced (P< 0. 05), but those in the low-concentration group and moderate-concentration group were not significantly different (both P>0. 05). Compared with the control group, the content of serum MDA in the low-concentration group, moderate-concentration group and high-concentration group was increased insignificantly (all P>0. 05). There were no significant differences regarding activities of SOD and content of MDA in lung organ and bronchoalveolar lavage fluid among each moxa smoke group (all P>0. 05). CONCLUSION: There is no obvious toxic reaction in the low-concentration group and moderate-concentration group; in the high-concentration group the antioxidant ability is damaged due to long-term exposure.

Current status of acupuncture and moxibustion in China.

Acupuncture and moxibustion are more integrated in the Chinese healthcare system than in the national healthcare systems of other countries. Development of acupuncture and moxibustion in China is making progress in this field. For overseas researchers, this commentary offers perspectives on the current status of acupuncture and moxibustion in China and examines relevant opportunities and challenges in healthcare reforms. There has been a steady increase in the number of undergraduates and postgraduates studying acupuncture and moxibustion in Chinese Medicine (CM) universities in China over the past decade. The legislation of CM physicians that was established in 1999 and the launch of continuing medical education in CM in 2002 have ensured the basic competency of practitioners. The Chinese Government has also shown support for CM development by increasing investment in related fields of research and administration. New challenges have emerged as the healthcare landscape in China has evolved over the past decade. It is important to harness the potential of acupuncture and moxibustion to create a value-driven healthcare system that meets the health needs of a rapidly aging society.