[Effect of moxibustion on autophagy in mice with Alzheimer's disease based on mTOR/p70S6K signaling pathway].

OBJECTIVE: To investigate the effect of moxibustion on autophagy and amyloid ß-peptide1-42 (Aß1-42) protein expression in amyloid precursor protein/presenilin 1 (APP/PS1) double-transgenic mice with Alzheimer's disease (AD). METHODS: After 2-month adaptive feeding, fifty-six 6-month-old APP/PS1 double transgenic AD mice were randomly divided into a model group, a moxibustion group, a rapamycin group and an inhibitor group, 14 mice in each group. Another 14 C57BL/6J mice with the same age were used as a normal group. The mice in the moxibustion group were treated with monkshood cake-separated moxibustion at "Baihui"(GV 20), "Fengfu" (GV 16) and "Dazhui" (GV 14) for 20 min; the mice in the rapamycin group were intraperitoneally injected with rapamycin (2 mg/kg); the mice in the inhibitor group were treated with moxibustion and injection of 1.5 mg/kg 3-methyladenine (3-MA). All the treatments were given once a day for consecutive 2 weeks. The morphology of hippocampal tissue was observed by HE staining; the ultrastructure of hippocampal tissue was observed by transmission electron microscopy; the expression of Aß1-42 protein in frontal cortex and hippocampal tissue was detected by immunohistochemistry; the expressions of mammalian target of rapamycin (mTOR), phosphorylated mTOR (p-mTOR), p70 ribosomal protein S6 kinase (p70S6K) and phosphorylated p70S6K (p-p70S6K) protein in hippocampus were detected by Western blot method. RESULTS: Compared with the normal group, the number of neuron cells was decreased, cells were necrotic and deformed, and autophagy vesicle and lysosome were decreased in the model group. Compared with the model group, the number of neuron cells was increased, cell necrosis was decreased, and autophagy vesicle and lysosome were increased in the moxibustion group and the rapamycin group. Compared with the normal group, the protein expressions of Aß1-42, mTOR, p-mTOR, p70S6K and p-p70S6K in the model group were increased (P<0.05); compared with the model group, the protein expressions of Aß1-42, mTOR, p-mTOR, p70S6K and p-p70S6K in the moxibustion group, rapamycin group and inhibitor group were decreased (P<0.05); compared with the inhibitor group, the protein expressions of Aß1-42, mTOR, p-mTOR, p70S6K and p-p70S6K in the moxibustion group and rapamycin group were decreased (P<0.05); compared with the rapamycin group, the protein expressions of mTOR, p-mTOR, p70S6K and p-p70S6K in the moxibustion group were decreased (P<0.05). CONCLUSION: Moxibustion could enhance autophagy in hippocampal tissue of APP/PS1 double transgenic AD mice and reduce abnormal Aß aggregation in brain tissue, the mechanism may be related to the inhibition of mTOR/p70S6K signaling pathway.

[Effect of moxibustion on cognitive function and proteins related with apoptosis of hippocampal neurons in rats with vascular dementia].

OBJECTIVE: To observe the effect of moxibustion on proteins related with apoptosis of hippocampal neurons in rats with vascular dementia (VD), and to explore the possible mechanism of moxibustion on improving VD. METHODS: Thirty SD rats were selected from 100 rats (3 rats were excluded) and randomly divided into a normal group and a sham operation group, 15 rats in each group. The remaining 67 rats were treated with ischemia-reperfusion method at bilateral common carotid artery to establish VD model. The 45 rats with successful VD model were randomly divided into a model group, a moxibustion group and a medication group, 15 rats in each group. On the 7th day after successful modeling, the rats in the moxibustion group were treated with suspended moxibustion at "Guanyuan" (CV 4), "Mingmen" (GV 4) and "Dazhui" (GV 14), 15 min per acupoint, once a day; there was 1 d of rest after 6 d of moxibustion, and the treatment was given for 4 weeks. The rats in the medication group was treated with nimodipine tablets by gavage, 2 mg/kg per day, 3 times a day for 4 weeks. Before and after intervention, the Morris water maze test was used to detect the escape latency of rats in each group; after the intervention, the TUNEL method was used to detect the apoptosis rate of neurons in hippocampal CA1 area; the immunofluorescence double labeling method was used to detect the number of co-expression positive cells of B-cell lymphoma-2 (Bcl-2)/neuronal nuclear antigen (NeuN) and Bcl-2-associated X protein (Bax)/NeuN in hippocampal CA1 area; the immunofluorescence single labeling method was used to detect cytochrome C (cytC) and outer mitochondrial membrane receptor Tom20 (Tom20) in hippocampal CA1 area; the Western blot method was used to detect the p53 upregulated modulator of apoptosis (PUMA) in hippocampus. RESULTS: Before intervention, compared with the normal group and the sham operation group, the escape latency in the model group, the moxibustion group and the medication group was prolonged (P<0.01). After intervention, the escape latency in the moxibustion group and the medication group was shorter than that before intervention (P<0.01). Compared with the model group, the escape latency in the moxibustion group and the medication group was shortened (P<0.05); compared with the medication group, the escape latency in the moxibustion group was shortened (P<0.05). Compared with the normal group and the sham operation group, the apoptosis rate of neurons in hippocampal CA1 area was increased, the number of Bcl-2/NeuN co-expression positive cells was decreased, and the number of Bax/NeuN co-expression positive cells was increased in the model group (P<0.01); compared with the model group, the apoptosis rates of hippocampal CA1 neurons were decreased, the number of Bcl-2/NeuN co-expression positive cells was increased, and the number of Bax/NeuN co-expression positive cells was decreased in the moxibustion group and the medication group (P<0.01); compared with the medication group, the apoptosis rate of neurons in hippocampal CA1 area was decreased, the number of Bcl-2/NeuN co-expression positive cells was increased, and the number of Bax/NeuN co-expression positive cells was decreased in the moxibustion group (P<0.01, P<0.05). Compared with the normal group and the sham operation group, the expressions of cytC, Tom20 protein in hippocampal CA1 area and PUMA protein in hippocampal tissue in the model group were increased (P<0.01); compared with the model group, the expressions of cytC, Tom20 protein in hippocampal CA1 area and PUMA protein in hippocampal tissue in the moxibustion group and the medication group were decreased (P<0.01); compared with the medication group, the expressions of cytC, Tom20 protein in hippocampal CA1 area and PUMA protein in hippocampal tissue in the moxibustion group were decreased (P<0.05, P<0.01). CONCLUSION: Moxibustion could improve the cognitive function of VD rats, which may be related to reducing the expression of Bax, cytC, Tom20 and PUMA protein in hippocampal CA1 area, promoting the release of Bcl-2 and inhibiting the apoptosis of hippocampal neurons.

[Acupuncture and moxibustion for vascular dementia and its effect on serum VEGF and AChE].

OBJECTIVE: To observe the efficacy of Huayu Tongluo moxibustion combined with acupuncture at Jing-well points on the differentiated meridians and routine acupuncture for vascular dementia (VD) and its effect on serum levels of vascular endothelial growth factor (VEGF) and acetylcholinesterase (AChE). METHODS: A total of 60 patients with VD were randomized into an observation group (30 cases, 1 case dropped off) and a control group (30 cases, 2 cases dropped off). In the observation group, Huayu Tongluo moxibustion combined with acupuncture at Jing-well points on the differentiated meridians and routine acupuncture were adopted, Huayu Tongluo moxibustion was applied at Baihui (GV 20), Dazhui (GV 14) and Shenting (GV 24); acupuncture at Jing-well points on the differentiated meridians was applied at corresponding Jing-well points according to pattern of syndrome; routine acupuncture was applied at Baihui (GV 20), Sishencong (EX-HN 1), etc. Acupuncture and moxibustion were given once a day, 6 times a week for 4 weeks. In the control group, donepezil was prescribed for oral administration, 5 mg each time, once a day for 4 weeks. Before and after treatment, the scores of mini mental state examination (MMSE) and activities of daily living (ADL) were observed, and the serum levels of AChE and VEGF were detected in the two groups. RESULTS: Compared before treatment, the MMSE and ADL scores and serum level of VEGF were increased (P<0.05), the serum level of AChE was decreased (P<0.05) after treatment in the two groups. The MMSE and ADL scores after treatment in the observation group were higher than the control group (P<0.05). CONCLUSION: Huayu Tongluo moxibustion combined with acupuncture at Jing-well points on the differentiated meridians and routine acupuncture could improve cognitive function and activities of daily living, which may be related to the regulation of serum levels of VEGF and AChE.

[Effect of moxibustion on expression of GAP-43 in sciatic nerve trunk and ventral horn of spinal cord of rats with primary sciatica and its nerve repair function].

OBJECTIVE: To observe the effect of moxibustion at "Huantiao" (GB 30) on the expression of growth-associated protein-43 (GAP-43) in the sciatic nerve trunk and ventral horn of spinal cord (L4-L6) in rats with primary sciatica, and to explore the mechanism of moxibustion in improving primary sciatica. METHODS: A total of 48 healthy male SD rats were randomly divided into a normal group, a sham operation group, a model group and a moxibustion group, 12 rats in each group. The rat model of primary sciatic pain was established by chronic constriction injury (CCI) of the sciatic nerve in the model group and the moxibustion group. On the 8th day of the experiment, moxibustion was adopted at "Huantiao" (GB 30) in the moxibustion group for 5-10 min, once a day for 14 consecutive days. Sciatic nerve function index (SFI) was measured and compared in each group at day 1, 7, 14 and 21. On the 21st day of the experiment, HE staining was used to observe the morphology of ventral horn of rat spinal cord and sciatic nerve trunk. Immunohistochemical method and real-time PCR were used to detect mRNA and protein expressions of GAP-43 in the spinal cord and sciatic nerve trunk of rats. RESULTS: On day 7, 14 and 21, there was no statistical difference in SFI between the sham operation group and the normal group (P>0.05); compared with the sham operation group on day 7, 14 and 21, the SFI of the model group was reduced (P<0.01); compared with the model group on day 14 and 21, SFI in the moxibustion group was increased (P<0.01). In the normal group and the sham operation group, neuronal cells were in order in the ventral horn of the spinal cord, nissl bodies were spaced regularly, the myelin sheath structure of sciatic nerve axon was clearly visible. In the model group, neuronal cells were deformed and ruptured in the ventral horn of the spinal cord, the number of nissl bodies was less, and the demyelination of sciatic axons appeared. In the moxibustion group, neuronal cells were found in the ventral horn of spinal cord, and the number of nissl bodies was increased, and less demyelinating changes of axons appeared in sciatic nerve. Compared with the normal group, the expressions of GAP-43 mRNA and GAP-43 protein in the sciatic nerve trunk and GAP-43 protein in the ventral horn of spinal cord were increased in the sham operation group (P<0.01). Compared with the sham operation group, the expression of GAP-43 mRNA and GAP-43 protein in the spinal cord and sciatic nerve trunk of rats in the model group was increased. Compared with the model group, the expression of GAP-43 mRNA and GAP-43 protein in the spinal cord and sciatic nerve trunk of rats in the moxibustion group was increased (P<0.01). CONCLUSION: Moxibustion at "Huantiao" (GB 30) could improve the sciatic nerve function in rats with primary sciatica and its mechanism may be related to improving the expression of GAP-43 and enhancing the self-repair ability of the sciatic nerve after injury.

[Effect of moxibustion on learning-memory ability and expression of hippocampal inflammatory factors and microtubule associated proteins in vascular dementia rats].

OBJECTIVE: To observe the effect of moxibustion on learning-memory ability and expression of hippocampal inflammatory factors and microtubule-associated protein doublecortin (DCX, a marker of neuronal regeneration) in vascular dementia (VD) rats, so as to explore its mechanisms underlying improvement of VD. METHODS: SD rats were randomly divided into normal control, sham operation, VD model, moxibustion and medication groups (n=15 rats in each group). The VD model was established by repeated occlusion of the bilateral common carotid arteries and reperfusion. Moxibustion was applied to "Guanyuan" (CV4), "Mingmen" (GV4) and "Dazhui"(GV14) for 15 min, once a day, 6 days a week for 4 weeks. Rats of the medication group were treated by gavage of Nimodipine (2mg·kg-1·d-1) 3 times daily for 4 weeks. Morris water maze test was used to detect the average escape latency of location navigation tasks for assessing the rats' learning-memory ability. H.E. staining was used to detect histopathological changes of the hippocampus tissue. The number of DCX-positive neurons (DCX/NeuN co-expression) in the dentate gyrus (DG) region of hippocampus was counted under microscope after immunofluorescence double staining, the immunoactivity of hippocampal DCX detected by using immunohistochemistry stain and the expression of DCX, TNF-α, IL-1ß, MPO, NF-κB p65 and IL-6 proteins in the hippocampus tissue detected using Western blot. RESULTS: Following modeling, the average escape latency was significantly longer in the model group than in the normal control and sham operation groups (P<0.01), and notably shorter in both the moxibustion and medication groups than in the model group after the treatment (P<0.01, P<0.05). The number of DCX-positive neurons, and the expression levels of DCX, TNF-α, IL-1ß, MPO, NF-κB p65 and IL-6 proteins in the hippocampus were significantly increased in the model group in comparison with the normal control and sham operation groups (P<0.01, P<0.05). After the interventions and in comparison with the model group, the number of DCX-positive neurons and the expression level of DCX were further up-regulated in both moxibustion and medication groups (P<0.01), while the expression levels of hippocampal TNF-α, IL-1ß, MPO, NF-κB p65 and IL-6 proteins were considerably down-regulated in the moxibustion and medication groups (P<0.01). The effect of moxibustion was weaker than that of medication in down-regulating the expression of TNF-α,MPO, NF-κB p65, IL-6 and IL-1ß, and in up-regulating DCX-positive neuron number and DCX expression (P<0.05, P<0.01). H.E. staining showed loose arrangement of neurons (with vague neuronal membrane in some cells), uneven organelle chromatin, disappearance of partial nucleolus, necrocytosis, and infiltration of small number of lymphocytes after modeling, which was relatively milder in both moxibustion and medication groups. CONCLUSION: Moxibustion can improve learning-memory ability in VD rats, which may be related to its effect in down-regulating the expression of inflammatory factors and up-regulating the expression of DCX to promote neuronal repair and regeneration.

[CAI Sheng-chao's experience in treatment of Sjögren's syndrome with moxibustion for promoting meridian circulation and warming yang].

Professor CAI Sheng-chao's clinical experience is summarized in the treatment of Sjögren's syndrome with moxibustion for promoting meridian circulation and warming yang and the typical case is introduced. Professor CAI believes that Sjögren's syndrome refers to zao (dry) bi syndrome. It is the insufficiency of yin and body fluid for the primary and yang qi deficiency for the root. On the base of traditional long-snake moxibustion, the moxibustion therapy for promoting meridian circulation and warming yang is adopted to warm up and tonify zangfu organs. When the function of yang qi is restored, body fluid (yin) will be generated naturally. Hence, the diagnosis and treatment system is developed by the integration of the theory as "moxibustion applicable for yin deficiency" and the characteristic moxibustion therapy.

[Moxibustion Improves Learning Ability by Regulating Hippocampal Neurotrophic Factor Expression and Notch Signaling in Vascular Dementia Rats].

OBJECTIVE: To observe the effect of moxibustion on the learning ability and expression of neurotrophic factors and Notch signaling in vascular dementia (VD) rats, so as to explore its neurogenesis mechanism underlying improvement of VD. METHODS: Sixty SD rats were equally and randomly divided into sham operation (sham), model, medication and moxibustion groups (n=15 rats/group). The VD model was established by occlusion of the bilateral cervical common arteries and reperfusion. Moxibustion was applied to "Dazhui"(GV 14), "Guanyuan"(CV 4) and "Mingmen"(GV 4)for 15 minutes, once daily, 6 times a week for 4 weeks. Rats of the medication group were treated by gavage of nimodipine (2 mg·kg-1·d-1),3 times a day, 6 days a week for 4 weeks. Morris water maze tests were performed to detect the rat's learning-memory ability. The infarcted size of the brain was detected by using 2,3,5-triphenyltetrazolium chloride (TTC) staining, and H.E. staining was used to detect the histopathological changes. The expression level of glia fibrillary acidic protein (GFAP), nerve growth factor (NGF), brain derived neurotrophic factor (BDNF), Notch 1 (a receptor), Hes 3 (a downstream effector) mRNAs and proteins in the hippocampal tissues were detected by quantitative real-time PCR and Western blot, respectively. RESULTS: After 4 weeks' intervention, modeling-induced increase of escape latency was significantly shortened in both moxibustion and medication groups relevant to the model group (P<0.05), and the infarct size was reduced and the damage degree of nerve cells in the brain tissue alleviated. The expression levels of BDNF, NGF, GFAP, Hes 3, Notch 1 genes and proteins were significantly up-regulated in the model group relevant to the sham operation group (P<0.05,P<0.01). After the intervention, the expression levels of hippocampal BDNF, NGF, GFAP, Hes 3 and Notch 1 mRNAs and proteins in the moxibustion group, and NGF and GFAP mRNAs, and BDNF, NGF, GFAP, Hes 3 and Notch 1 proteins in the medication group were further obviously up-regulated relevant to the model group (P<0.05, P<0.01). The effect of moxibustion was significantly superior to that of medication in up-regulating GFAP, Hes 3, Notch 1 mRNAs expression (P<0.05,P<0.01). No significant differences were found in the expression of BDNF, Hes 3 and Notch 1 mRNAs in the medication group relevant to the model group (P<0.05), and between the moxibustion and medication groups in up-regulating the expression of BDNF and NGF mRNAs, and in up-regulating the expression of BDNF, NGF, GFAP, Hes 3 and Notch 1 proteins (P<0.05). CONCLUSIONS: Moxibustion can improve learning ability in VD rats, which may be associated with its effects in up-regulating the expression of neurotrophic factors and in potentiating Notch signaling.

[Clinical study on primary osteoporosis treated with spreading moxibustion for warming yang and activating blood circulation].

OBJECTIVE: To observe the efficacy on primary osteoporosis treated with spreading moxibustion for warming yang and activating blood circulation so as to provide the effective clinical therapeutic methods for osteoporosis. METHODS: Sixty cases of primary osteoporosis were randomized into a spreading moxibustion group (30 cases) and a calcium tablet group (30 cases). In the calcium tablet group, caltrate was prescribed for oral administration, 600 mg per day. In the spreading moxibustion group, on the basis of the treatment as the calcium tablet group, the spreading moxibustion was applied at Dazhui (GV 14) to Yaoshu (GV 2) for warming yang and activating blood circulation. The duration of treatment was 12 weeks. Visual analogue scale (VAS) score, TCM clinical symptom score and bone mineral density (BMD) were observed and compared before and after treatment in the patients between the two groups. RESULTS: VAS scores were reduced apparently after treatment in the two groups (both P < 0.01) and the results in the spreading moxibustion group were obviously superior to that in the calcium tablet group (2.36 +/- 0.43 vs 4.52 +/- 0.35, P < 0.01). BMD were all increased in the two groups (P < 0.05, P < 0.01) and the results in the spreading moxibustion group were superior to those in the calcium tablet group (both P < 0.05). The total clinical effective rate was 86.67% (26/30) in the spreading moxibustion group, apparently better than 63.33% (19/30) in the calcium tablet group (P < 0.05). TCM clinical symptom scores after treatment were all reduced apparently in the two groups (both P < 0.01), and the result in the spreading moxibustion group was obviously superior to that in the calcium tablet group (4.72 +/- 1.90 vs 6.82 +/- 2.30, P < 0.01). The total effective rate of TCM symptoms was 93.33% (28/30) in the spreading moxibustion group, apparently better than 70.00% (21/30) in the calcium tablet group (P < 0.05). CONCLUSION: The combined therapy of spreading moxibustion for warming yang and activating blood circulation and the oral administration of caltrate apparently relieves pain and TCM clinical symptoms, improves BMD in the patients of osteoporosis and achieves definite clinical efficacy in the patients of osteoporosis.

[Discussion on twenty four moxibustion styles of Meihua acupuncture-moxibustion school].

Nomination, apparatus, manipulating techniques, indications and theoretical basis of 14 single moxibustion styles including Chuijiu (insufflating moxibustion), Dianjiubi Jiu (pecking moxibustion with a pen-like stool), Jiujia Xunjiu (moxibustion with frame), Tongmai Wenyang Jiu (moxibustion for removing meridian obstructions and warming up yang), QifuJiu (moxibustion on umbilicus and abdomen), Xiongyang Jiu (moxibustion on the chest for reinforcing yang qi), Toujing Jiu (moxibustion on head and neck), Anmo Jiu (moxibustion with massage), Zhiti Jiu (moxibustion on extremities), Guan Jiu (moxibustion with a tube), Zu Jiu (moxibustion on foot), Wenzhenjiu (warm needling), Huanong Jiu (festering moxibustion) and Gewu Jiu (indirect moxibustion) are expounded in this article. And 10 compound moxibustion with the combination of 2 or more than 2 above mentioned single moxibustion style under the instruction of combination of local and distal points, combination of upper and lower points as well as combination of frontal and back points are also stated. It suggests to classify moxibustion into categories of festering moxibustion and mild moxibustion, indirect moxibustion and direct moxibustion, and to classify moxibustion apparatus into the categories of treating tools and assisting tools.

[Clinical research on cake-separated moxibustion for impaired glucose regulation].

OBJECTIVE: To observe clinical curative effect of cake-separated moxibustion on impaired glucose regulation (IGR) and explore its action mechanism. METHODS: Sixty cases were randomly divided into a simple lifestyle intervention group (control group) and a cake-separated moxibustion combined with lifestyle intervention group (observation group), 30 cases in each one. The control group was treated with lifestyle intervention. Based on lifestyle intervention, cake-separated moxibustion at Pishu (BL 20), Weishu (BL 21) and Yishu (EX-B 3) was applied to the observation group. Fast plasma glucose (FPG), two hours plasma glucose after oral glucose tolerance test (OGTT2hPG), fasting insulin (FINS), homa insulin resistance index (HOMA-IR), blood lipid, body mass index (BMI) and waist circumference (WC) were observed in the two groups before and after treatment. RESULTS: After treatment, the OGTT2hPG and FPG were both decreased significantly (both P<0.05) in the two groups, compared between the two groups, the differences of FPG [(0.41 +/- 0.42) mmol/L vs (0.05 +/- 0.08)mmol/L] and OGTT2hPG [(0.85 +/- 0.53)mmol/L vs (0.17 +/- 0.19)mmol/L] were both statistically significant. There were no significant changes in FINS, HOMA-IR, blood lipid, BMI and WC in the control group before and after treatment (all P>0.05), but FINS, HOMA-IR levels, triglycerides (TG), total cholest-erol (TC), low density lipoprotein (LDL-C), BMI and WC in the observation group were decreased obviously after treatment (all P<0.05), which had statistical differences between the two groups (all P<0.05). CONCLUSION: The cake-separated moxibustion combined with lifestyle intervention can obviously control blood glucose levels, improve insulin resistance and blood lipid levels, decrease BMI and WC.

[Contribution of Zhou Mei-sheng to cause of acupuncture and moxibustion].

Chief physician zhou Mei-sheng, a famous expert of acupuncture and moxibustion in our country, has made outstanding contribution to development of cause of acupuncture and moxibustion in over half a century, including paying attention to practice, concentrating attention on clinical study and creation of acupuncture and moxibustion instruments; being at good summarization, workers with original view, resolving doubts, medical skill and medical ethics passing on to later generations; believing moxibustion able to treat AIDS. In recent over 20 years, he devotes study on acupuncture and moxibustion for treatment of difficult and complicated diseases and writes books to expound a theory.