Improved Suspension Fixation Using Rat Jacket in Rat Acupuncture Experiments.

In basic experimental acupuncture research, rats are commonly used as laboratory animals. However, it is difficult for them to maintain a fixed posture. During electroacupuncture procedures, proper immobilization of rats is essential. Various methods of rat fixation are currently used, including anesthesia fixation, high-platform fixation, binding fixation, and fixation with a self-made rat coat. However, these methods have their limitations, which may affect the efficiency and operability of the experiment to some extent. This protocol introduces a method of suspending and fixing rats using rat clothes. Firstly, rats are clothed with rat jackets that match their body shape, taking advantage of their preference for darkness and burrowing. The needling operation can then be carried out after the rats have worn rat clothes. When suspended, the rats are relatively still, as their limbs cannot move. This fixation method offers not only economical and user-friendly benefits but also ensures a stable and reliable fixation of the rats in a comfortably relaxed position. It also effectively minimizes time consumption, experimental space, and manpower resources. Additionally, this method allows for the exposure of most acupoints used for acupuncture in rats. This article primarily concentrates on the device's composition, encompassing a specially designed rat jacket, an elevated fixation rack, and their connecting structures. Additionally, an illustrative example will be presented to demonstrate the application of the rat clothing-based suspension fixation method in rat electroacupuncture procedures.

[Protective mechanism of salvianolic acid B on blood vessels].

Salvianolic acid B(Sal B) is the main water-soluble component of Salvia miltiorrhiza Bunge. Studies have found that Sal B has a good protective effect on blood vessels. Sal B can protect endothelial cells by anti-oxidative stress, inducing autophagy, inhibiting endoplasmic reticulum stress(ERS), inhibiting endothelial inflammation and adhesion molecule expression, inhibiting endothelial cell permeability, anti-thrombosis, and other ways. In addition, Sal B can alleviate endothelial cell damage caused by high glucose(HG). For vascular smooth muscle cell(VSMC), Sal B can reduce the synthesis and secretion of inflammatory factors by inhibiting cyclooxygenase. It can also play a vasodilatory role by inhibiting Ca~(2+) influx. In addition, Sal B can inhibit VSMC proliferation and migration, thereby alleviating vascular stenosis. Sal B also inhibits lipid deposition in the subendothelium, inhibits macrophage conversion to foam cells, and reduces macrophage apoptosis, thereby reducing the volume of subendothelial lipid plaques. For some atherosclerosis(AS) complications, such as peripheral artery disease(PAD), Sal B can promote angiogenesis, thereby improving ischemia. It should be pointed out that the conclusions obtained from different experiments are not completely consistent, which needs further research. In addition, previous pharmacokinetics showed that Sal B was poorly absorbed by oral administration, and it was unstable in the stomach, with a large first-pass effect in the liver. Sal B had fast distribution and metabolism in vivo and short drug action time. These affect the bioavailability and biological effects of Sal B, and the development of clinically valuable Sal B non-injectable delivery systems remains a great challenge.

[Electroacupuncture inhibits neuron injury of SAMP8 mice by reducing inflammatory response].

OBJECTIVE: To observe the effect of electroacupuncture (EA) at "Baihui"(GV20) and "Shenshu" (BL23) on the pathological injury of neurons in SAMP8 mice and the anti-inflammatory effect on neuron repair, providing a new experimental basis for EA prevention and treatment of Alzheimer's disease. METHODS: Twelve 7-month-old SAMP8 mice were randomly divided into model and EA groups, and 6 SAMR1 mice of the same age and genetic background were used as normal group. Mice in the EA group were needled at GV20 and bilateral BL23, and EA (1 mA, 2 Hz) was applied to bilateral BL23 for 15 min, once daily, 10 d as a course for a total of 4 courses, with an interval of 1 d. Mice in the normal and model groups were captured and fixed in the same way as the EA group. The spatial learning and memory ability was detected by Morris water maze test. Neuronal nuclear antigen (NeuN) positive expression and the number of NeuN-positive cells in dentate gyrus (DG) were detected by immunofluorescence staining. The protein expression levels of ionized calcium binding adapter molecule 1(Iba-1), tumor necrosis factor-α(TNF-α), interleukin (IL)-6 and IL-1ß in hippocampus were detected by Western blot. The ultrastructure of nerve cells in DG was observed by transmission electron microscopy. RESULTS: Compared with the normal group, the average escape latency was prolonged(P<0.01), the number of platform crossing was significantly reduced (P<0.01), the average fluorescence intensity of NeuN and the number of NeuN-positive cells in hippocampus DG region decreased (P<0.05), the expression levels of Iba-1, TNF-α, IL-6 and IL-1ß in hippocampus were increased (P<0.05) in the model group.Compared with the model group, the ave-rage escape latency was shortened (P<0.01), the number of platform crossing times was significantly increased (P<0.01), the average fluorescence intensity of NeuN and the number of NeuN-positive cells in hippocampus DG region increased (P<0.05), the expression levels of Iba-1, TNF-α, IL-6 and IL-1ß in hippocampus were decreased (P<0.05) in the EA group. The morphology of nerve cells in the hippocampus DG region was normal， and the organelles in the cytoplasm were clear, complete and regularly distributed in the normal group. However, the morphology of nerve cells in the model group was seriously irregular, which was also irregular in EA group but somewhat relieved compared with model group. CONCLUSION: EA at GV20 and BL23 can improve the learning and memory ability of SAMP8 mice, which may be related to inhibiting the neuroinflammatory response, increasing the number of neurons and improving the ultrastructure of the DG region of the hippocampus to play the role of neuron protection.

Ancient Chinese Herbal Recipe Huanglian Jie Du Decoction for Ischemic Stroke: An Overview of Current Evidence.

Ischemic stroke is a major cause of mortality and neurological morbidity worldwide. The underlying pathophysiology of ischemic stroke is highly complicated and correlates with various pathological processes, including neuroinflammation, oxidative stress injury, altered cell apoptosis and autophagy, excitotoxicity, and acidosis. The current treatment for ischemic stroke is limited to thrombolytic therapy such as recombinant tissue plasminogen activator. However, tissue plasminogen activator is limited by a very narrow therapeutic time window (<4.5 hours), selective efficacy, and hemorrhagic complication. Hence, the development of novel therapies to prevent ischemic damage to the brain is urgent. Chinese herbal medicine has a long history in treating stroke and its sequela. In the past decades, extensive studies have focused on the neuroprotective effects of Huanglian Jie Du decoction (HLJDD), an ancient and classical Chinese herbal formula that can treat a wide spectrum of disorders including ischemic stroke. In this review, the current evidence of HLJDD and its bioactive components for ischemic stroke is comprehensively reviewed, and their potential application directions in ischemic stroke management are discussed.

Hydraulic conductivity of novel geosynthetic clay liner to bauxite liquor from China: Modified fluid loss test evaluation.

A modified sodium bentonite geosynthetic clay liner (GCL) designed for acid-and-alkaline resistance was evaluated for its potential application in the containment of bauxite residue leachate. A modified fluid loss test was employed to quickly evaluate the hydraulic conductivity (k) of the GCL using distilled water, tap water, and four bauxite liquors (BLs, leachate from bauxite residue reservoirs). The effects of swelling capacity of bentonite, prehydration, hydraulic gradient (i), ionic strength (I), and relative abundance of monovalent and multivalent cations (RMD) on the hydraulic conductivity of the GCL were analyzed. The results indicated that the BLs significantly decreased free swell index of the bentonite. As compared to increasing i, prehydration obviously enhanced hydraulic performance of the GCL. The four BLs increased k of the GCL by a factor of 4-12 relative to the tap water permeation condition, and the resultant k exceeded upper limit of 5.0 × 10-11 m/s for GCLs. The increase in k was attributed to compression in diffuse double layer of the bentonite and dissolution in clay minerals in ion-rich and hyperalkaline BLs, manifesting that further modification on the GCL is needed. The I was found a better indicator than the RMD on correlation with chemical compatibility of the GCL.

Comparison of limestone calcined clay cement and ordinary Portland cement for stabilization/solidification of Pb-Zn smelter residue.

Decreasing carbon emissions by replacing Portland cement (PC) with supplementary cementitious materials (SCMs), such as low-grade limestone (LS) and calcined clays (CC), has tremendous potential for stabilization/solidification (S/S) of industrial hazardous waste primarily with heavy metals. Recently, a low-carbon-based cementitious binder, namely, limestone calcined clay cement (LC), has emerged as an alternative for S/S treatment of wastes. However, comprehensive comparison between LC and PC application in solidifying/stabilizing wastes has not been conducted. This study aims to investigate the S/S efficiency of Pb-Zn smelter residue (LZSR) comprising heavy metals lead (Pb), zinc (Zn), and cadmium (Cd) at higher concentrations. LZSR is treated with LC and PC for capturing strength and leaching toxicity. The test results indicate that low-grade CC and LS in the LC binder can promote the alkaline environment, and act as fillers in solidifying heavy metals. The toxicity characteristic leaching procedure leaching concentrations of untreated (UT) LZSR were 503 mg/kg, 1266 mg/kg, and 251 mg/kg for Pb, Zn, and Cd, respectively. After a 28-day curing, the leaching concentrations in LC-treated LZSR reduced to 4.33 mg/kg, 189.68 mg/kg, and 0.46 mg/kg, while the leaching concentrations of PC-treated LZSR reduced to 29 mg/kg, 338 mg/kg, and 6 mg/kg for Pb, Zn, and Cd, respectively. The maximum immobilization efficiencies for Pb, Zn, and Cd reached 85%, 99%, and 99%, respectively. Moreover, the insoluble phases for Pb, Zn, and Cd obtained from the sequential extraction test results were 63.5%, 72.1%, and 42.4% for LC-treated LZSR and 35.7%, 38%, and 43% for PC-treated LZSR with binder content of 8% binder and curing time of 28 days. Increasing curing time and binder content reduced leaching concentrations, and the underneath mechanisms were interpreted by XRD, SEM-EDS, and FTIR analyses. Overall, the results indicate that Pb, Zn, and Cd can be successfully immobilized using 8% LC binder by transforming soluble heavy metals to insoluble hydroxides and their complexes.

Mechanism of Action of Acupuncture in Obesity: A Perspective From the Hypothalamus.

Obesity is a prevalent metabolic disease caused by an imbalance in food intake and energy expenditure. Although acupuncture is widely used in the treatment of obesity in a clinical setting, its mechanism has not been adequately elucidated. As the key pivot of appetite signals, the hypothalamus receives afferent and efferent signals from the brainstem and peripheral tissue, leading to the formation of a complex appetite regulation circuit, thereby effectively regulating food intake and energy homeostasis. This review mainly discusses the relationship between the hypothalamic nuclei, related neuropeptides, brainstem, peripheral signals, and obesity, as well as mechanisms of acupuncture on obesity from the perspective of the hypothalamus, exploring the current evidence and therapeutic targets for mechanism of action of acupuncture in obesity.

Preventive electroacupuncture reduces cognitive deficits in a rat model of D-galactose-induced aging.

Acupuncture can reduce cognitive deficits in Alzheimer's disease. However, whether electroacupuncture can prevent or alleviate the cognitive deficits in animal models of aging remains poorly understood. Studies have shown that disordered epigenetic modifications play a critical role in age-related cognitive decline. Therefore, we hypothesized that preventive electroacupuncture might improve cognitive functions during aging by regulating epigenetic modifications. A rat model of aging was produced by intraperitoneal injection of 120 mg/kg D-galactose for 8 weeks. Baihui and Shenshu acupoints were stimulated by electroacupuncture for 8 weeks from the first day of D-galactose administration. Preventive electroacupuncture alleviated memory impairment, decreased tau hyperphosphorylation, and reduced glycogen synthase kinase-3ß protein and mRNA expression levels in the brainstem dorsal raphe nucleus, where intracellular neurofibrillary tangle lesions first occur. In addition, the DNA methylation level in the promoter region of the glycogen synthase kinase-3ß gene was increased. The effects of preventive electroacupuncture were stronger than those of preventive acupuncture. Intraperitoneal injection of 0.4 mg/kg 5-aza-2'-deoxycytidine, an inhibitor of DNA methyltransferase that blocks epigenetic modifications, antagonized the effects of preventive electroacupuncture. Our results suggest that preventive electroacupuncture treatment alleviates cognitive impairment in aging rats probably by affecting the epigenetic modification of the glycogen synthase kinase-3ß gene in the dorsal raphe nucleus. This study was approved by the Animal Ethics Committee of Hubei University of Chinese Medicine, China (approval No. HUCMS201712001) on November 28, 2017.

Preventive Electroacupuncture Ameliorates D-Galactose-Induced Alzheimer's Disease-Like Pathology and Memory Deficits Probably via Inhibition of GSK3ß/mTOR Signaling Pathway.

Acupuncture has been practiced to treat neuropsychiatric disorders for a thousand years in China. Prevention of disease by acupuncture and moxibustion treatment, guided by the theory of Chinese acupuncture, gradually draws growing attention nowadays and has been investigated in the role of the prevention and treatment of mental disorders such as AD. Despite its well-documented efficacy, its biological action remains greatly invalidated. Here, we sought to observe whether preventive electroacupuncture during the aging process could alleviate learning and memory deficits in D-galactose-induced aged rats. We found that preventive electroacupuncture at GV20-BL23 acupoints during aging attenuated the hippocampal loss of dendritic spines, ameliorated neuronal microtubule injuries, and increased the expressions of postsynaptic PSD95 and presynaptic SYN, two important synapse-associated proteins involved in synaptic plasticity. Furthermore, we observed an inhibition of GSK3ß/mTOR pathway activity accompanied by a decrease in tau phosphorylation level and prompted autophagy activity induced by preventive electroacupuncture. Our results suggested that preventive electroacupuncture can prevent and alleviate memory deficits and ameliorate synapse and neuronal microtubule damage in aging rats, which was probably via the inhibition of GSK3ß/mTOR signaling pathway. It may provide new insights for the identification of prevention strategies of AD.

Experimental Evidence of the Benefits of Acupuncture for Alzheimer's Disease: An Updated Review.

As the global population ages, the prevalence of Alzheimer's disease (AD), the most common form of dementia, is also increasing. At present, there are no widely recognized drugs able to ameliorate the cognitive dysfunction caused by AD. The failure of several promising clinical trials in recent years has highlighted the urgent need for novel strategies to both prevent and treat AD. Notably, a growing body of literature supports the efficacy of acupuncture for AD. In this review, we summarize the previously reported mechanisms of acupuncture's beneficial effects in AD, including the ability of acupuncture to modulate Aß metabolism, tau phosphorylation, neurotransmitters, neurogenesis, synapse and neuron function, autophagy, neuronal apoptosis, neuroinflammation, cerebral glucose metabolism, and brain responses. Taken together, these findings suggest that acupuncture provides therapeutic effects for AD.

New ternary blend limestone calcined clay cement for solidification/stabilization of zinc contaminated soil.

This study evaluates the potential use of a new limestone calcined clay cement (LC3) for stabilization/solidification of zinc contaminated soil. LC3 is a new ternary blend manufactured by the replacement of 50% cement clinker by locally available two supplementary cementitious materials (SCMs) - limestone and calcined clay. The incorporation of LC3 is evaluated on the soil spiked with 0.5% and 1% of Zinc (Zn) at curing times of 3, 7, 14, 28 and 56 days. pH, strength and leachability properties of the solidified/stabilised soil are measured for both mechanical and environmental conditions. Additionally, sequential extraction procedure (SEP), X-ray diffraction (XRD) analysis and scanning electron microscope (SEM) analysis are performed to elucidate the mechanisms of Zn immobilization in the soil. The results show that the leachable Zn concentrations in the stabilised soil are well below the corresponding hazardous waste management regulatory limit after the curing time of 14 days. The soil pH and unconfined compressive strength of the stabilised soil increase with curing time. The SEP results confirm that LC3 considerably reduces the acid soluble fraction (F1) and increase the residual fraction (F4). The XRD and SEM results indicate that formation of Tri-calcium Silicate 3CaO·SiO2, Portlandite Ca(OH)2, Ettringite Ca6Al2(SO4)3(OH)12.26 H2O and Wulfingite Zn(OH)2 are the primary mechanisms for the immobilization of Zn in the LC3 stabilised soil.

Microbial induced carbonate precipitation for immobilizing Pb contaminants: Toxic effects on bacterial activity and immobilization efficiency.

Microbial induced carbonate precipitation (MICP) is a natural bio-mediated process, which has been explored for soil stabilization and heavy metals immobilization in soil and groundwater. Previous studies have shown that MICP is capable of immobilizing various heavy metals including lead (Pb). However, most studies focus merely on the immobilization of heavy metals with relatively low concentration. This study: (1) presents results of an investigation into the toxic effects of Pb on bacterial activity and immobilization efficiency within a wide range of Pb concentrations; and (2) identifies controlling biotic and abiotic factors of Pb immobilization by MICP. In the first series of tests, bacterial strains (Sporosarcina pasteurii) are inoculated into nutrient solutions containing 0-50 mM Pb(NO3)2 and incubated at 30 °C. Biochemical parameters are measured over time, which include pH, electrical conductivity, urease activity, and viable cell number. In the second series of tests, grown bacterial strains are mixed with urea, calcium salts and Pb(NO3)2 in solution. Viable cell number, produced ammonium concentration, aqueous Pb concentration of the mixed solution, and total precipitation mass are measured. The results show that the presence of Pb has marginal effect on bacterial growth and associated urease activity at Pb concentration < 30 mM. The calcium source and initial bacteria concentration are found to remarkably influence Pb immobilization efficiency in terms of Pb removal percentage. Supplementary geochemical simulation results indicate that the Pb immobilization mechanisms includes abiotic precipitation, biotic precipitation and bio-sorption.

[Acupuncture plus moxibustion intervention improves learning-memory ability by suppressing hippocampal JAK2/STAT3 signaling in Alzheimer's rats].

OBJECTIVE: To investigate the effect of acupuncture plus moxibustion on learning-memory ability and expression of hippocampal Janus kinase-2 (JAK2)/signal transducer and activator of transcription-3 (STAT3)/suppressors of cytokine signaling-3 (SOCS3) signaling in Alzheimer's Disease (AD) rats, so as to reveal their mechanisms underlying improvement of AD. METHODS: A total of 60 SD rats were randomly divided into four groups：normal control, sham-operation, model and acupuncture-moxibustion (Acu-moxi, n＝15 in each group) groups. The AD model was established by microinjection of ß-amyloid 1-42(Aß1-42，5 µL)into the bilateral hippocampus. Seven days after modeling, Acu-moxi intervention was given. After insertion of acupuncture needles into "Baihui" (GV20) and bilateral "Shenshu" (BL23) and manipulating them for a while, the needles were then retained for 15 min, when, the mild moxibustion was performed at the same time. The treatment was conducted once daily, 5 times a week for consecutive 4 weeks. After the treatment, Morris water maze test was used to detect the animals' learning-memory ability. Immunohistochemistry and Western blot were respectively used to detect the number of positive cells and protein expression levels of JAK2, STAT3 and SOCS3 in the hippocampus tissue. RESULTS: Following modeling and compared with the normal control and sham-operation groups, the average escape latency was significantly prolonged (P<0.01), and the number of the original platform crossing and the residence time in the platform quadrant were significantly shortened in the model group (P<0.01). The numbers of hippocampal JAK2- and STAT3-positive cells and expression levels of hippocampal JAK2 and STAT3 proteins were significantly increased (P<0.01), and the number of hippocampal SOCS3-positive cells as well as the expression of SOCS3 protein significantly decreased in the model group relevant to the normal control and sham-operation groups (P<0.01). After the intervention, the average escape latency was significantly shortened (P< 0.01), and the number of the original platform crossing and the residence time in the platform quadrant were significantly increased in the Acu-moxi group (P<0.01), and the expression levels of JAK2 and STAT3 were significantly down-regulated and that of SOCS3 was considerably up-regulated in the Acu-moxi group relevant to the model group (P<0.01)．. CONCLUSION: Acu-moxi intervention can improve the learning-memory ability in AD rats, which is associated with its functions in inhibiting hippocampal JAK2/STAT3 signaling and up-regulating SOCS3 (a negative feedback factor) protein level.

[Effect of acupuncture on the ultrastructure of neurons and astrocytes in the hippocampal dentate gyrus in rats with Alzheimer's disease induced by Aß1-42].

OBJECTIVE: To observe the effects of acupuncture at "Baihui" (GV 20) and "Shenshu" (BL 23) on the ultrastructure of hippocampal dentate gyrus in rats with Alzheimer's disease. METHODS: Forty SPF Wistar male rats were randomly divided into a normal group, a sham operation group, a model group and an acupuncture group, 10 rats in each one. The rats in the model group and the acupuncture group were treated with injection of 5 µL Aß1-42 at bilateral hippocampus, while the rats in the sham operation group were treated with injection of 5 µL 0.9% NaCl. Three days after modeling, the rats in the acupuncture group were treated with acupuncture at "Baihui" (GV 20) and "Shenshu" (BL 23) for 20 min, once a day, six treatments constituted a course, and totally two courses were given with an interval of 1 day between courses. The rats in the other groups received normal diet and no treatment was given. Before modeling, four days after modeling and after treatment, water maze test was performed to observe the escape latency and the number of crossing platforms. The hippocampal dentate gyrus was collected and transmission electron microscope was applied to observe the ultrastructure changes of neurons and astrocytes. RESULTS: ①Four days after modeling, compared with the normal group and the sham operation group, the escape latency was significantly prolonged and the number of crossing platforms was reduced in the model group (all P<0.01); after treatment, compared with the model group, the escape latency was significantly reduced and the number of crossing platforms was increased in the acupuncture group (both P<0.01). ②In the normal group and the sham operation group, the morphology of neurons and astrocytes was intact, the nuclear and membrane structure were clear, and the morphology of organelles such as mitochondria, endoplasmic reticulum and lysosomes was normal. In the model group, the morphology of neurons was irregular, the nucleus was severely constricted with edema in the cytoplasm, the color of heterochromatin was deepened, the endoplasmic reticulum was expanded, the granulation was removed and the number of mitochondria was decreased, even with malformed-like change in mitochondrial cristae; there was severe edema around astrocytes, few organelles in the cytoplasm, severe swelling of mitochondria and mild expansion of the endoplasmic reticulum. In the acupuncture group, the edema of the neuron and astrocytes was still evident, and the mitochondrial was mildly swollen but relieved compared with that in the model group, and there were no obvious abnormalities in neuronal endoplasmic reticulum and ribosomes. CONCLUSION: Acupuncture could improve the ultrastructure of neurons and astrocytes in the hippocampal dentate gyrus in rats with Alzheimer's disease induced by Aß1-42.

[Acupuncture and Moxibustion Improves Learning-memory Ability of Alzheimer's Disease Rats Possibly by Up-regulating Serum Aß Internalization Enzyme Contents].

OBJECTIVE: To observe the effect of acupuncture and moxibustion (AM) on learning-memory ability and expression of amyloid beta (Aß) in the hippocampal dentate gyrus (DG) of Alzheimer's disease (AD) rats, so as to explore its mechanism underlying improvement of AD. METHODS: Forty male Wistar rats were randomly divided into normal, sham operation, model and AM groups (n＝10 in each). The AD model was established by bilateral hippocampal injection of Aß1-42(5 µL). The AM was applied at "Baihui" (GV 20) and "Shenshu" (BL 23) for 15 min, once daily for 12 times. Morris water maze tests were used to assess the rats' learning-memory ability. The levels of serum Aß1-42 and Aß internalizing enzymes including transthyretin (TTR), lipoprotein lipase (LPL), alpha 2 macroglobulin (α 2M) and apolipoprotein E (ApoE) were detected by ELISA. The expression of Aß1-42 in the hippocampal DG was detected by immunohistochemistry. RESULTS: Compared with the sham operation group, the average escape latency of location navigation test was significantly prolonged in the first 5 days and the last 3 days (P<0.01), and the residence time of the target-quadrant and the platform crossing times of special probing task were significantly reduced in the model group (P<0.01). After the intervention, the average escape latency was obviously shortened in the first 5 days and the last 3 days (P<0.01), and both the residence time and the platform crossing times were significantly increased in the AM group relevant to the model group (P<0.01, P<0.05). After modeling, the contents of TTR, LPL, α 2M and ApoE in the serum were significantly lower in the model group than in the sham operation group (P<0.01, P<0.05), while the levels of serum Aß1-42 and hippocampal Aß1-42 expression were considerably increased relevant to the sham operation group (P<0.01). Following the intervention, the TTR, LPL, α2M and ApoE contents in the serum were significantly increased (P<0.01, P<0.05), and both serum Aß1-42 and hippocampal Aß1-42 expression were notably down-regulated in comparison with the model group (P<0.05). No significant differences were found between the normal and sham operation groups in the levels of all the above-mentioned indexes (P>0.05). CONCLUSION: AM can improve the learning-memory ability of AD rats, which may be related to its effects in up-regulating the contents of serum Aß internalizing enzymes and promoting the clearance of hippocampal Aß. It suggests a protective role of AM on hippocampal neurons.

[Effect of "Kidney-reinforcing and Govenor Vessel-regulating" of Acupuncture plus Moxibustion on Mitochondrial Dynamics-related Proteins in Hippocampal Neurons of Rats with Alzheimer's Disease].

OBJECTIVE: To observe the effect of acupuncture plus moxibustion on mitochondrial dynamics-related proteins (OPA 1, Fis 1) of hippocampal neurons in rat with Alzheimer's disease (AD) , and to explore the potential mechanism underlying acupuncture for the protection of hippocampal neuronal mitochondria. METHODS: Eighty male Wistar rats were randomly divided into normal, sham-operation, model and acu-moxibustion groups. AD model was made by injecting Aß1-42 into the hippocampus. "Baihui" (GV 20) and "Shenshu" (BL 23) acupoints were stimulated by manual acupuncture (15 min) in combination with moxibustion (15 min) for two sessions of treatment, with seven consecutive treatments for one session and one day break between the two sessions. The expression of hippocampal OPA 1 and Fis 1 was assessed by means of Western blot and immunofluorescence technique. RESULTS: The expression level of OPA 1 and Fis 1 of rats in the model group was significantly lower and higher, respectively, than those of the normal and sham-operation groups (P < 0.01). Acupuncture plus moxibustion intervention notably increased OPA 1 and decreased Fis 1 expression levels in the acu-moxibustion group compared to those of the model group (P < 0.01). CONCLUSION: Acupuncture combined with moxibustion produce benefit to hippocampal neurons in AD rats probably via the regulation of the mitochondrial dynamics (imbalance between fission and fusion proteins) and improvement of the mitochondrial damage of hippocampus.

[Protective mechanism of acupuncture-moxibustion on hippocampal neuron mitochondria in rats with Alzheimer's disease].

OBJECTIVE: To observe effects of acupuncture and moxibustion on ultrastructure and silent information regulator 1 (SIR1) in hippocampal neuron mitochondria in rats with Alzheimer's disease (AD) in order to explore its possible effective mechanism during the process of protecting mitochondria. METHODS: Eighty Wistar rats were randomly divided into a normal group, a sham operation group, a model group and an acupuncture group, 20 rats in each one. The AD model was established in the model group and acupuncture group with injection of Amyloid beta Protein Fragment1-14 (Abeta1-42) into the hippocampus. Acupuncture and moxibustion was applied at "Baihui" (GV 20) and "Shenshu" (BL 23) in the acupuncture group, once a day, seven days as a treatment course and totally two course were required. The rest groups were all fed with normal diet, without any treatment. The transmission electron microscopy, immunohistochemistry and immunoblotting technique were respectively adapted to measure ultrastructure and level of STR1 in hippocampal neuron mitochondria in AD rats. RESULTS: Compared with the model group, the ultrastructure in hippocampal neuron mitochondria in the acupuncture group was effectively improved. The average optical density of SIR1 in hippocampus was 0.21 +/- 0.10 and the gray value was 136.82 +/- 47.42 in the model group, which were obviously lower than 0.47 +/- 0.09 and 281.44 +/- 57.98 in the normal group (both P < 0.01). However, levels of SIR1 in the acupuncture group, they were 0.32 +/- 0.11 and 199.52 +/- 58.12, which were significantly increased compared with those in the model group (both P < 0.05). CONCLUSION: The reinforcing-kidney and regulating-Governor Vessel method of acupuncture and moxibustion for AD could improve ultrastructure of mitochondria and increase levels of SIR1 to achieve the aim of recovering injury of mitochondria and protecting function of mitochondria.

[Effects of acupuncture and moxibustion on energy metabolism-related protein of hippocampal neuron mitochondria in Alzheimer's disease rats].

OBJECTIVE: To explore action mechanism of acupuncture and moxibustion for Alzheimer's disease (AD) to provide evidence for prevention and treatment with acupuncture and moxibustion on AD in clinic. METHODS: Eighty SPF-grade male Wistar rats, (200 +/- 20) g, were randomly divided into a normal group, a sham-operation group, a model group and a treatment group, 20 cases in each one. The model was duplicated with injection of Abeta1-42 in rats' hippocampus. Expect the treatment group, the rest groups were treated with regular feeding after respective intervention. The treatment group was treated with acupuncture and moxibustion at "Baihui" (GV 20) and "Shenshu" (BL 23), once a day, seven days as a treatment course and totally for two courses. There was one day of interval between the courses. The immunohistochemistry and quantitative RT-PCR methods were applied to test level of Abeta-binding alcohol dehydrogense (ABAD) and cytochrome oxidase IV (COX IV) in hippocampal neurons mitochondria. RESULTS: Acupuncture and moxibustion could reduce effectively level of ABAD and improve activity of COX IV in hippocampal neurons mitochondria in the treatment group, which has statistical significance compared with that in the model group (P < 0.01) and no statistical significance compared with that in the normal group and sham-operation group (P > 0.05). This indicated that acupuncture and moxibustion could effectively suppress overexpression of ABAD, improve activity of COX IV and reduce leak of reactive oxygen species, which could improve metabolic disturbance of mitochondria energy to achieve the goal of prevention and treatment of AD. CONCLUSION: The prevention and treatment of AD with acupuncture and moxibustion could be related with suppressing overexpression of ABAD and improving activity of COX IV in hippocampal neurons mitochondria to improve mitochondria energy metabolism.

[Effects of electroacupuncture on the expression of GDNF and Ret in Parkinson's disease model rats].

OBJECTIVE: To explore the mechanism of electroacupuncture therapy on Parkinson's disease (PD). METHODS: Fifty Wistar rats were randomly divided into a normal group, a sham-operation group, a model group, a Fengfu-Taichong group and a Shuanggu Yitong group. PD model was duplicated by microinjection of 6-Hydroxyl-Dopamine into right corpora striata, and by microinjection of normal saline in sham-operation group. Rats in normal group, sham-operation group and model group were not treated. In Fengfu-Taichong group, the rats were treated by electroacupuncture at "Fengfu" (GV 16) and "Taichong" (LR 3) on the basis of the PD model, and by electroacupuncture at "Fengfu" (GV 16), "Taichong" (LR 3), "Guanyuan" (CV 4) and "Zusanli" (ST 36) in Shuanggu Yitong group, once daily for 2 weeks. GDNF and Ret expression were detected by immunohistochemistry and western blotting, respectively. RESULTS: The number of GDNF positive cells and the content of Ret receptor increased significantly in the two electroacupuncture groups compared with those in the other groups (all P < 0.01), and the expression of GDNF increased significantly in Shuanggu Yitong group compared with that in Fengfu-Taichong group (P < 0.01). CONCLUSION: Electroacupuncture can not only increase the expression of GDNF, but also enhance its effect. "Shuanggu Yitong" method is better than simple acupuncture at "Fengfu" (GV 16) and "Taichong" (LR 3) in increasing expression of GDNF.

[Effects of acupoint-embedment of medicated-thread and acupoint-injection on expression of cortical urokinase-type plasminogen activator and plasminogen activator inhibitor-1 in rats with cerebral ischemia-reperfusion injury].

OBJECTIVE: To observe the effect of acupoint-embedement of medicated-thread and acupoint-injection of Chuanxiongzine on the expression of urokinase-type plasminogen activator (uPA) and plasminogen activator inhibitor-1 (PAI-1) in the cerebral cortex in rats with cerebral ischemia-reperfusion injury (CI/RI), so as to explore its underlying mechanism in protecting the ischemic cerebral tissue. METHODS: Seventy-eight SD rats were randomly divided into normal control group (n=6), sham operation (sham) group (n=18), model group (n=18),acupoint injection (Al) group (n=18), and acupoint-thread-embedment (ATE) group (n=18). Rats of the latter 4 groups were randomized into 1 d, 3 d and 5 d subgroups, with 6 rats in each. CI/RI model was established by occlusion of the right middle cerebral artery (MCAO) for 30 min and reperfusion. For rats of the Al group, Chuanxiongzine (0.1 mL/200 g) was injected into "Baihui" (GV 20) and "Dazhui" (GV 14), and for those of ATE group, a piece of medicated thread containing collagen protein (extracted from the rat's tail tissue) and Chuanxiongzine + retarder was embedded into GV 20 and GV 14, respectively. The expression of uPA and PAI-1 in the cerebral cortex on the ischemia side was detected by immunohistochemistry. RESULTS: In comparison with the normal control group, the expression of uPA of the ischemia cerebral cortex on day 1, 3 and day 5 in the model group was increased significantly (P < 0.01), while the PAI-1 expression decreased remarkably in the model group (P < 0.01). Compared with the 3 time-points of the model group, cortical uPA expression levels at the 3 time-points in the Al group and those of day 3 and day 5 in the ATE group were down-regulated significantly (P < 0.01), whereas cortical PAI-1 expression levels at the 3 time-points in both AI and ATE groups up-regulated considerably (P < 0.05, P < 0.01). Comparison between AI and ATE groups showed that the expression levels of cortical uPA in the latter group on day 3 and day 5 were significantly lower than those of the former group (P < 0.05), whereas the cortical PAI-1 expression levels in the latter group on day 3 and day 5 were evidently higher than those of the former group (P < 0.05). But, cortical PAI-1 expression of the ATE group on day 1 was significantly lower than that of the AI group (P < 0.05). No significant differences were found between the AI and ATE groups in the expression level of cortical uPA on day 1 and between normal and sham groups in both uPA and PAI-1 expression levels at the 3 time-points (P > 0.05). CONCLUSION: Both AI and ATE can down-regulate cortical uPA expression and up-regulate cortical PAI-1 expression in rats with CI/RI, which may contribute to their protective effect in reducing cerebral ischemic injury.