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Objectives: To observe the effect of electroacupuncture (EA) at "Neiguan"(PC6) on cardiac function, cardiac morphology and transient receptor potential channel (TRPC) protein expressions in myocardial tissue of mice with myocardial hypertrophy, so as to explore its mechanisms underlying improvement of myocardial hypertrophy., Methods: Forty-five male C57BL/6 mice were randomly divided into control, model and EA groups (15 mice/group). The myocardial hypertrophy model was established by subcutaneous injection of isoproterenol hydrochloride (15 mg·kg -1 ·d -1 ) for 14 days. The mice of the control group received subcutaneous injection of same amount of normal saline. The mice of the EA group received EA stimulation (frequency of 2 Hz, intensity of 1 mA) of bilateral PC6 for 20 min each time, once a day for 14 consecutive days. After the intervention, the body weight, tibia length and heart weight were measured. The left ventricular ejection fraction (EF), fractional shortening index (FS), left ventricular end-systolic volume (LVEV), left ventricular end-systolic internal diameter (LVID) and left ventricular posterior wall thickness (LVPW) were measured by using echocardiography for evaluating the cardiac function. The mean number and surface area of myocardial cells was detected by wheat germ agglutinin (WGA) staining, and changes of the cardiac morphology were observed under light microscopy after HE staining. The expression levels of TRPC1, TRPC3, TRPC4 and TRPC6 (TRPC1/3/4/6) in the myocardial tissue were detected by real-time quantitative PCR (qPCR) and Western blot, separately., Results: Compared with the control group, the heart-body weight ratio( P <0.05) and heart-weight-to-tibia-length ratio ( P <0.01), LVEV and LVID levels, the relative surface area, left ventricular area ratio, and the expression levels of cardiac TRPC1/3/4/6 were significantly increased ( P <0.01, P <0.05), while the EF, FS, LVPW, number of cardiomyocytes, and the left ventricular posterior wall ratio were obviously decreased ( P <0.01, P <0.05) in the model group. In comparison with the model group, the heart/body weight ratio, heart-weight-to-tibia-length ratio, LVEV and LVID levels, relative surface area, left ventricular area ratio, and the expression levels of cardiac TRPC1/3/4/6 were significantly decreased ( P <0.01, P <0.05), while the EF, FS, LVPW, number of cardiomyocytes and left ventricular posterior wall ratio were significantly increased ( P <0.01, P <0.05) in the EA group. H.E. staining showed disordered arrangement of cardiomyocytes and obvious myocardial interstitial inflammatory cell infiltration in the model group, and evident reduction of degree of cardiac fibrosis and interstitial edema in the EA group., Conclusions: EA of PC6 can improve the cardiac function and cardiac morphology in mice with myocardial hypertrophy, which may be related to its functions in down-regulating the expression of transient receptor potential channels.

A moxibustion device with the functions of auricular fumigation moxibustion and heat-sensitive moxibustion is designed. The smoke of the ignited moxa stick is used for the fumigation moxibustion at the external auditory canal, while the heat generated works on Dazhui (GV 14) for heat-sensitive moxibustion. The device consists of five parts, i.e. combustion chamber, smoke pipe, smoke processing chamber, power module and connector. It solves the limitations such as unpleasant experience in treatment, unfavorable temperature control, easy scalding and excessive manual dependence induced by usual fumigation moxibustion and during heat-sensitive moxibustion. This moxibustion device may improve the safety and convenience when delivering the treatment with fumigation moxibustion and heat-sensitive moxibustion, as well as the work efficiency of medical staff.

The paper summarizes the definition, location and main indication of qicimai points recorded in Huangdi Neijing ( Yellow Emperor 's Inner Classic ). It is found that qicimai points are the "upward moving points" in reference to the meridian distribution rule of "rooting, running, infusing and moving"; and corresponding to the sites of "running outwards and inwards" of the meridians' "separating, meeting and running outwards and inwards". It also includes the infusing points for the sea of qi and marrow. The new idea, "selecting qicimai points for the treatment of qi obstruction in the neck gate", is proposed. Based on the systematic application of the acupoints on the nape region, it is anticipated that a new approach will be provided to the treatment of the diseases in the neck, shoulder, head, face, the five sensory organs, mental disorders and zangfu qi dysfunction.

Electroacupuncture (EA) is well documented to treat irritable bowel syndrome (IBS). However, the mechanism of the central nervous system related to IBS and acupuncture stimulation is still not well known. In this study, a rat model of IBS was established by cold-restraint comprehensive stresses for 15 days, and it was found that the levels of corticotropin-releasing hormone (CRH), corticosterone (CORT), and adrenocorticotropic hormone (ACTH) in the peripheral serum were increased; the visceral sensitivity was enhanced; and the intestinal motility was accelerated, specifically, there was an enhancement in the discharge frequency of neurons in the paraventricular nucleus (PVN). EA treatment for 3 days, 20 min/day, alleviated the increase in the levels of CRH, CORT, and ACTH in the peripheral serum of rats, reduced the visceral sensitivity of IBS rats, and inhibited colon movement and discharge frequency of the neurons in the PVN. In addition, EA could reduce the excitability of CRH neurons and the expression of corticotropin-releasing hormone receptor 1 (CRHR1) and corticotropin-releasing hormone receptor 2 (CRHR2) in PVN. At the same time, the expression of CRH, CRHR1, and CRHR2 in the peripheral colon was decreased. Taken together, EA appears to regulate intestinal functional activity through the central CRH nervous system, revealing the central regulation mechanism of EA in IBS rats, and providing a scientific research basis for the correlation among the meridians, viscera, and brain. NEW & NOTEWORTHY The purpose of this research was to determine the central regulatory mechanism of electroacupuncture (EA) in rats with irritable bowel syndrome (IBS). Our results showed that combined with the serum changes in corticotropin-releasing hormone (CRH), corticosterone (CORT), and adrenocorticotropic hormone (ACTH), the improvement of IBS by EA was related to them. Furthermore, EA could regulate intestinal functional activity through the central CRH + nervous system.

An automatic ash-removal heat-sensitive moxibustion device was developed, which could keep relatively constant temperature of heat-sensitive moxibustion, and realize the automatic ignition and automatic ash removal of moxa sticks during heat-sensitive moxibustion. The automatic ash-removal heat-sensitive moxibustion device comprises a bracket and a moxibustion box fixed on the top of the bracket; the bracket is composed of a base and a movable telescopic arm. This device can solve the problems of temperature instability, moxa ash blocking heat transfer and moxa ash falling during heat-sensitive moxibustion, avoiding the scalding caused by moxa ash falling, and reduce the workload of medical staff.

Objective: To observe the effect of moxibustion on the expressions of hypoxia inducible factor-1α (HIF-1α) and vascular endothelial growth factor (VEGF) in ankle synovial tissue of rats with adjuvant arthritis(AA), so as to explore the mechanism of moxibustion in inhibiting synovial angiogenesis and improving joint symptoms of rheumatoid arthritis., Methods: Sixty healthy male SD rats were randomly divided into normal group, model group, moxibustion group and medication group, with 15 rats in each group. AA rat model was established by subcutaneous injection of Freund's complete adjuvant into the right hind paw. Rats in the moxibustion group were treated with "Zusanli" (ST36), "Guanyuan" (CV4) and "Ashi" point moxibustion, 20 min each time, once a day, for consecutive 3 weeks. Rats in the medication group were given methotrexate (0.35 mg/kg) intragastric administration, twice a week, for consecutive 3 weeks. Foot plantar volume of rats was measured by toe volume mea-suring instrument. HE staining was used to observe the histopathology of ankle synovium. The protein expressions of HIF-1α and VEGF in ankle synovial tissue were detected by immunohistochemistry and Western blot., Results: Compared with the normal group, the foot plantar volume and the protein expressions of HIF-1α and VEGF in synovial tissue of ankle joint were significantly increased ( P <0.01) in the model group, the synovial tissue showed obvious hyperplasia and a large number of neovasculogenesis. Following the interventions, the foot plantar volume and the protein expressions of HIF-1α and VEGF in synovial tissue of ankle joint were significantly decreased ( P <0.05, P <0.01) in both moxibustion and medication groups in contrast to the model group, and there was no obvious proliferation of synovial tissue, and only a few neovascularization was observed. Compared with the medication group, the foot plantar volume was decreased ( P <0.05) in the moxibustion group., Conclusion: Moxibustion can improve joint swelling and inhibit synovial angiogenesis in AA rats, and its mechanism may be related to down-regulating of HIF-1α and VEGF protein expressions.

Objective: To observe the effects of electroacupuncture (EA) on cardiac function and local field potential (LFP) in sensory and motor cortices in mice with stress cardiomyopathy (SC), and to explore the possible mechanism of EA in improving SC., Methods: Twenty-seven female C57BL/6 mice were randomized into a blank group, a model group and an EA group, 9 mice in each group. In the model group and the EA group, SC model was established by continuous intraperitoneal injection of isoproterenol (ISO) for 14 days. At the same time of modeling, EA was applied at "Neiguan" (PC 6) and "Shenmen" (HT 7) in the EA group, with disperse-dense wave, in frequency of 2 Hz/15 Hz, 15 min each time, once a day for 14 days. After intervention, the total movement distance, the number of crossing grid and the number of crossing central grid of open field test within 5 minutes were observed; the left ventricular function indexes (left ventricular diameter of end-diastole [LVIDd], left ventricular diameter of end-systole [LVIDs], left ventricular volume of end-diastole [LVEDV], left ventricular volume of end-systole [LVESV], ejection fraction [EF] and fraction shortening [FS]) were detected by echocardiography; the changes in ST-segment amplitude and PR interval of electrocardiogram were observed; the morphology of myocardial tissue was observed by HE staining; the serum levels of cortisol (CORT), cardiac troponin T (cTnT) and brain natriuretic peptide (BNP) were detected by ELISA; the changes of LFP in sensory and motor cortices were recorded by Plexon multi-channel acquisition system., Results: Compared with the blank group, in the model group, the total movement distance, the number of crossing grid and the number of crossing central grid of open field test were decreased ( P <0.05); LVIDd, LVIDs, LVEDV and LVESV were increased ( P <0.05), EF and FS were decreased ( P <0.05); ST-segment amplitude was increased ( P <0.05) and PR interval was prolonged ( P <0.05); irregular myocardial fiber arrangement, interstitial edema and inflammatory cell infiltration were observed; the serum levels of CORT, cTnT and BNP were increased ( P <0.05); in the sensory cortex, the ratios of delta, theta, alpha and beta frequency bands were increased ( P <0.05), the maximum energy spectrum of theta and beta frequency bands was increased ( P <0.05), the power spectral density (PSD) of delta, theta, alpha, beta and gamma frequency bands was increased ( P <0.05); in the motor cortex, the ratios of delta, theta, alpha and beta frequency bands were increased ( P <0.05), the maximum energy spectrum as well as PSD of delta, theta, alpha, beta and gamma frequency bands were increased ( P <0.05). Compared with model group, in the EA group, the total movement distance, the number of crossing grid and the number of crossing central grid of open field test were increased ( P <0.05); LVIDd, LVIDs, LVEDV and LVESV were decreased ( P <0.05), EF and FS were increased ( P <0.05); ST-segment amplitude was decreased ( P <0.05), and the PR interval was shortened ( P <0.05); myocardial fiber injury and inflammatory cell infiltration were reduced; the serum levels of CORT, cTnT and BNP were decreased ( P <0.05); in the sensory cortex, the ratios of theta, alpha and beta frequency bands were decreased ( P <0.05), the ratio of gamma frequency band was increased ( P <0.05), the maximum energy spectrum of theta frequency band as well as the PSD of theta, alpha, beta and gamma frequency bands were decreased ( P <0.05); in the motor cortex, the ratios of theta, alpha and beta frequency bands were decreased ( P <0.05) and the ratio of gamma frequency band was increased ( P <0.05), the maximum energy spectrum of delta frequency band was increased ( P <0.05), the maximum energy spectrum of theta frequency band as well as the PSD of theta and gamma frequency bands were decreased ( P <0.05)., Conclusion: EA can improve cardiac function in mice with stress cardiomyopathy, and its mechanism may be related to the regulation of local field potentials in sensory and motor cortices.

Objective: To explore the effects of electroacupuncture(EA) on metabolic patterns of the prefrontal cortex in rats with acute myocardial ischemia., Methods: Eighteen SD rats were randomly divided into sham group, model group and EA group, with 6 rats in each group. Rats in the model and EA groups were subject to acute myocardial ischemia by ligation of the left anterior descending coronary artery. For the EA group, EA stimulation (1 mA, 2 Hz/15 Hz, 30 min) was applied to "Shenmen"(HT7) -"Tongli"(HT5) once a day for 3 consecutive days. The histopathological changes of myocardial tissue and levels of ischemia modified albumin (IMA) in serum were determined by HE staining and ELISA, respectively. The LC-MS/MS technique was used to characterize the metabolic profiling of the prefrontal cortex. The differentially expressed metabolites were screened by principal component analysis(PCA) and partial least squares-linear discriminant analysis (PLS-LDA), and subsequently Kyoto encyclopedia of genes and genomes (KEGG) metabolic pathway enrichment analysis was performed., Results: Compared with the sham group, the myocardial fibers were disordered and fractured, and content of serum IMA was significantly increased in the model group ( P <0.01), which, however, were significantly decreased in the EA group ( P <0.01). With PCA and PLS-LDA, there were 18 differential metabolites between the model and sham groups. Forty-eight differential metabolites were emerged between the EA and model groups. Three metabolites associated to the sphingolipid metabolism were reversed by EA stimulation, as indicated by KEGG., Conclusion: The molecular mechanism of EA against myocardial ischemia is partially mediated by regulating sphingolipid-related metabolites in the prefrontal cortex.

Objective: To observe the effect of electroacupuncture (EA) on vascular endothelial growth factor-C (VEGF-C), vascular endothelial growth factor receptor-3 (VEGFR-3), proinflammatory factors and apoptosis in myocardial tissue in mice with acute myocardial ischemia (AMI), and to explore the mechanism of EA for AMI., Methods: Fifty male C57BL/6 mice were randomly divided into a sham operation group, a model group, an EA group, an inhibitor group and an inhibitor+EA group, 10 mice in each group. Except for the sham operation group, the mice in the remaining groups were intervented with ligation at the left anterior descending (LAD) coronary artery to establish AMI model. The mice in the sham operation group were intervented without ligation after thoracotomy. The mice in the EA group were intervented with EA at "Shenmen" (HT 7) and "Tongli" (HT 5), disperse-dense wave, 2 Hz/15 Hz in frequency, 1 mA in current intensity, 30 min each time, once a day, for 3 d. The mice in the inhibitor group were treated with intraperitoneal injection of SAR 131675 (12.5 mg•kg -1 •d -1 , once a day for 3 d). The mice in the inhibitor+EA group were injected intraperitoneally with SAR 131675 30 min before EA. The ECG before modeling, 30 min after modeling and 3 d after intervention was detected, and the ST segment displacement was recorded; after the intervention, the ELISA method was applied to measure the contents of serum creatine kinase isoenzyme (CK-MB), aspartate aminotransferase (AST) as well as tumor necrosis factor-α (TNF-α) and interleukin-23 (IL-23) in myocardial tissue; the HE staining method was used to observe the morphological changes of myocardial tissue; the immunofluorescence double labeling method was applied to measure the number of co-expression positive cells of VEGF-C/VEGFR-3 in myocardial tissue; the TUNEL method was used to detect the level of cardiomyocyte apoptosis; the Western blot method was applied to measure the protein expressions of VEGF-C, VEGFR-3, b-lymphoma-2 (Bcl-2), activated caspase-3 (Cleaved Caspase-3) and activated poly adenosine diphosphate ribose polymerase-1 (Cleaved PARP-1)., Results: Compared with the sham operation group, in the model group the ST segment displacement was increased ( P <0.01); the contents of CK-MB, AST, TNF-α and IL-23 were increased ( P <0.01); the arrangement of myocardial fibers was disordered, and interstitial inflammatory cell infiltration was obvious; the number of co-expression positive cells of VEGF-C/VEGFR-3 was decreased ( P <0.01); the number of cardiomyocyte apoptosis was increased ( P <0.01); the expressions of VEGF-C, VEGFR-3 and Bcl-2 were decreased ( P <0.01); the expressions of Cleaved Caspase-3 and Cleaved PARP-1 were increased ( P <0.01). Compared with the model group, in the EA group the ST segment displacement was decreased ( P <0.01); the contents of CK-MB, AST, TNF-α, IL-23 were decreased ( P <0.01); the severity of myocardial pathological injury was reduced; the number of co-expression positive cells of VEGF-C/VEGFR-3 was increased ( P <0.01); the number of cardiomyocyte apoptosis was reduced ( P <0.01); the expressions of VEGF-C, VEGFR-3 and Bcl-2 were increased ( P <0.01); the expressions of Cleaved Caspase-3 and Cleaved PARP-1 were reduced ( P <0.01). There was no significant difference in all the indexes between the model group and the inhibitor group ( P >0.05). Compared with the model group, the protein expression of VEGF-C was increased in the inhibitor+EA group ( P <0.01). Compared with the inhibitor group, in the EA group the ST segment displacement was decreased ( P <0.01); the contents of CK-MB, AST, TNF-α, IL-23 were decreased ( P <0.01); the severity of myocardial pathological injury was reduced; the number of co-expression positive cells of VEGF-C/VEGFR-3 was increased ( P <0.05); the number of cardiomyocyte apoptosis was reduced ( P <0.01); the expressions of VEGF-C, VEGFR-3 and Bcl-2 were increased ( P <0.01); the expressions of Cleaved Caspase-3 and Cleaved PARP-1 were reduced ( P <0.01). Compared with the inhibitor+EA group, all the indexes in the EA group were improved except the protein expression of VEGF-C ( P <0.01)., Conclusion: EA could relieve the inflammatory reaction and apoptosis in AMI mice, and its mechanism may be related to activating VEGF-C/VEGFR-3 pathway and promoting lymphangion genesis.

Objective: To observe the effect of electroacupuncture (EA) on the cardiac function, lymphatic markers, macrophage and inflammatory cytokines in acute myocardial ischemia (AMI) mice, so as to explore its mechanism in improving AMI., Methods: Male C57BL/6 mice were randomly divided into sham operation, model, EA, inhibitor and inhibitor+EA groups, with 10 mice in each group. AMI model was established by occlusion of left anterior descending coronary artery. For mice in the EA group and inhibitor+EA group, EA (1 mA, 2 Hz/15 Hz) was applied to bilateral "Shenmen"(HT7) and "Tongli"(HT5) for 30 min, once daily for consecutive 3 days. Mice in the inhibitor+EA group were given intraperitoneal injection of vascular endothelial growth factor receptor-3 (VEGFR-3) inhibitor SAR131675 30 min before the EA, while mice in the inhibitor group were given intraperitoneal injection of SAR131675 only. The electrocardiogram (ECG) of the neck-thoracic lead was recorded and analyzed by BL-420F biological function experiment system. Histopathologic changes of myocardial tissue were observed after H.E. staining. The contents of lactate dehydrogenase (LDH), cardiac troponin I (cTnI) in serum and interleukin-18 (IL-18) and interleukin-6 (IL-6) in ischemic myocardium were determined by ELISA. The expressions of hyaluronic acid receptor-1 (LYVE-1) and macrophage mar-ker CD68 (CD68) in the myocardial tissue were detected by immunofluorescence assay. The protein expression levels of vascular endothelial growth factor C (VEGF-C) and VEGFR-3 were detected by Western blot., Results: Compared with the sham operation group, the ECG-ST level, the contents of serum LDH and cTnI, and the contents of IL-18 and IL-6 in the myocardial tissue were significantly increased ( P <0.01), the expression of LYVE-1 and the protein expression levels of VEGF-C and VEGFR-3 in the myocardial tissue were significantly decreased ( P <0.01), while the number of CD68 positive cells was significantly increased ( P <0.01) in the model group. Compared with the model, inhibitor and inhibitor+EA groups, the ECG-ST level, the contents of serum LDH and cTnI, and the contents of IL-18 and IL-6 in the myocardial tissue were decreased ( P <0.01), the expression of LYVE-1 and the expression level of VEGFR-3 protein were increased ( P <0.01), while the number of CD68 positive cells was significantly decreased ( P <0.01) in the EA group. Compared with the model and inhibitor groups, the expression level of VEGF-C was increased ( P <0.01) in the EA group. Outcomes of H.E. staining showed that the myocardial fibers were disordered with a large number of inflammatory cell infiltration in the model group, which was milder in the EA group., Conclusion: Acupuncture can improve the inflammatory injury of AMI mice, which may be related to activate VEGF-C/VEGFR-3 pathway to promote lymphangiogenesis, reduce macrophage infiltration and inflammatory factors.

Objective: To observe the effect of acupuncture on endoplasmic reticulum calcium, apoptosis number and Caspase-12 protein expression in hippocampal neurons of convulsive rats, so as to explore its mechanisms underlying improvement of convulsion., Methods: SD rats were randomly divided into normal control, model and acupuncture groups, with 36 rats in each group. Rats in the normal control group received intraperitoneal injection (i.p.) of normal saline (2 mL), and those of the other 2 groups received i.p. of pentylenetetrazole (50 mg/kg) for establishing convulsion model. Manual acupuncture stimulation was applied to "Baihui"(GV20) and "Dazhui"(GV14) for 30 min after modeling. The hippocampal tissues were taken at 2, 12 and 48 h after modeling. The endoplasmic reticulum Ca 2+ concentration (optical density, OD) was detected by using fluorescence probe technique and laser confocal microscopy, and the number of apoptosis of hippocampal neurons at the 3 time points detected by using terminal-deoxynucleoitidyl transferase mediated nick end labeling (TUNEL) stain. The expression of Caspase-12 protein in hippocampus at 3 time points was observed by immunohistochemistry., Results: In comparison with the normal group, the number of apoptotic cells of hippocampal neurons and the expression levels of Caspase-12 protein in hippocampus at 2, 12 and 48 h after seizures were obviously increased ( P <0.01), and the OD value of Ca 2+ at 3 time points significantly decreased ( P <0.01) in the model group.Following acupuncture intervention, the increased levels of the number of apoptotic cells of hippocampal neurons and the expression of Caspase-12 protein in hippocampus at 3 time points and the decreased levels of OD value of Ca 2+ at 3 time points were reversed in the acupuncture group ( P <0.05, P <0.01)., Conclusion: Acupuncture intervention is effective in reducing the apoptosis of hippocampal neurons in convulsion rats, which may be related to its functions in down-regulating Caspase-12 expression and promoting influx of Ca 2+ in the hippocampal neurons.

Objective: To observe the effect of acupuncture serum on the expression of microtubule associated protein-2 (MAP-2) and nerve growth associated protein-43 (GAP-43) in cultured hippocampal neurons of convulsive rats., Methods: The acute convulsion model was induced by intraperitoneal injection of pentylenetetrazol in SD rats who were then randomized into model group and acupuncture group. Rats of the acupuncture group received manual acupuncture stimulation of "Baihui" (GV20) and "Dazhui" (GV14) for 30 min, once daily for 7 days. Then, the blood samples taken from the abdominal aorta of rats in the convulsion model and acupuncture groups were processed into serum samples, i.e. non-acupuncture serum and acupuncture se-rum. The primary-cultured hippocampal neurons of fetal rats were cultured for 10 days and then divided into normal extracellular fluid (normal) group, magnesium (Mg 2+ ) free extracellular fluid group, acupuncture serum group and non-acupuncture serum group. At the 10th day, the neurons in the normal group were cultured continuously in extracellular fluid for 3 h, and then cultured in DMEM/F12(1∶1) medium (planting fluid); neurons in the Mg 2+ free group were cultured in magnesium-free fluid medium to induce epileptic-like discharge; neurons in the acupuncture serum group were cultured in the mixed medium of planting fluid and 10% acupuncture serum; and neurons in the non-acupuncture serum were cultured in the mixed culture medium of planting fluid and non-acupuncture serum (10%). At last, these neurons in the above-mentioned groups were cultured in the magnesium-free extracellular fluid continuously for 2, 12 and 48 h, respectively, followed by detecting the expression levels of MAP-2 and GAP-43 proteins at the 3 time points by using immunofluorescence and Western blot, separately., Results: The rate of MAP-2 positive cells and protein expression at 2, 12 and 48 h, and the rate of GAP-43 positive cells and protein expression at 12 and 48 h in the hippocampal neurons were significantly down-regulated in the Mg 2+ free group in contrast to the normal group ( P <0.05， P <0.01). Compared to the Mg 2+ free group, the rates of MAP-2 and GAP-43 positive cells and protein expression at 2, 12 and 48 h were considerably up-regulated in the acupuncture serum group ( P <0.05， P <0.01), but not in the non-acupuncture serum group (P>0.05)., Conclusion: Acupuncture serum can significantly up-regulate the expression of MAP-2 and GAP-43 proteins in hip-pocampal neurons, which may play a positive role in improving synaptic plasticity and neuronal damage in convulsion rats.

Objective: To explore the molecular mechanism of locus coeruleus（LC） involved in electroacupuncture (EA) anti myocardial ischemia., Methods: Twenty-four SD rats were randomly divided into sham-operation, model, EA and EA +lesion groups, with 6 rats in each group. The acute myocardial ischemia (AMI) model was established by ligation of the left anterior descending branch of coronary artery. EA (2 Hz/15 Hz, 1 mA) was applied to bilateral "Shenmen" (HT7) -"Tongli" (HT5) and the middle-point between HT7 and HT5 for 30 min, once daily for 3 days. For rats of the EA +lesion group, the virus (300 nL) was injected into bilateral LC before EA treatment. Serum aspartate aminotransferase (AST) was detected by ELISA. The gene expression profiles of rat heart were detected by transcriptome sequencing, the differentially expressed genes were screened, and Gene Ontology (GO) functional classification and Kyoto Encyclopedia of genes and genomes (KEGG) metabolic pathway enrichment analysis were performed., Results: Compared with the sham-operation group, serum AST content was significantly increased in the model group ( P <0.01). Following the intervention, serum AST was significantly reduced in the EA group ( P <0.01), while the serum AST in the EA + lesion group was significantly higher compared with the EA group ( P <0.05). Differential expression analysis showed that 1 138 differentially expressed genes were screened out between the model group and the sham-operation group, 1 330 differentially expressed genes between model and EA group, and 804 differentially expressed genes between EA and EA + lesion group. Among them, 218 differential genes were involved in the regulation of EA anti-myocardial ischemia in LC. GO functional classification analysis showed that these differentially expressed genes mainly involved in cell processes, metabolic processes and biological regulation in biological processes. KEGG pathway analysis showed that these differentially expressed genes were enriched in sulfur relay system, thiamine metabolism, glutathione metabolism, C5 branch dicarboxylic acid metabolism, cell adhesion molecules and Th1 and Th2 cell differentiation., Conclusion: EA intervention has a positive effect in anti-myocardial ischemia, which may be related to the sulfur relay system, thiamine metabolism, glutathione metabolism, C5 branch dicarboxylic acid metabolism, cell adhesion molecules and Th1 and Th2 cell differentiation involved in LC.

Focusing on the original text record in Huangdi Neijing ( Inner Canon of Yellow Emperor ), the relevant theories of "Tianyou (TE 16) and five regions" are explored, e.g. acupoint names, meridians and acupoint features, and the clinical application of these acupoints has been analyzed. It is discovered that "Tianyou (TE 16) and five regions" are mainly used in treatment of the disorders in the nervous system, five sensory organs and motor system. Besides, in terms of the relevant theories, "Tianyou (TE 16) and five regions" has been compared with "root and knot" and "twelve divergent meridians". It is found that "Tianyou (TE 16) and five regions" communicates the externally-internally related meridians and is applicable in treatment of the disorders with both exterior and interior involved. It is the essential acupoint composition of the human body.

Objective: To observe the effect of electroacupuncture (EA) at "Zusanli"(ST36) on the expression of epidermal growth factor receptor (EGFR), tumor necrosis factor α(TNF-α) transfer growth factor α(TGF-α), interleukin-8(IL-8), p38 mitogen-activated protein kinases (p38MAPK), mucin-5AC (MUC5AC) and other related factors in chronic obstructive pulmonary disease (COPD) rats, so as to reveal its underlying mechanisms in improving COPD., Methods: A total of thirty male SD rats were randomly divided into normal control, model and EA groups, with 10 rats in each group. The COPD model was replicated using a combined method of tracheal infusion of lipopolysaccharide (LPS) and forced smoke-inhaling. EA (1-3 mA, 4 Hz/20 Hz) was applied to bilateral ST36 for 30 min, once daily for two consecutive weeks. The lung ventilation activities including the forced vital capacity (FVC) and forced expiratory volume (FEV) at 0.1 and 0.3 s (FEV0.1, FEV0.3) were detected. Histopathological changes of the middle lobe and bronchus of the right lung were observed after H.E. staining. The contents of TGF-α, TNF-α and IL-8 in the serum, bronchoalveolar lavage fluid (BALF) and superior lobe of the right lung were assayed by using ELISA, and the expression levels of EGFR, p38MAPK and MUC5AC proteins (inferior lobe of the left lung) and mRNAs (inferior lobe of the right lung) detected using Western blot, immunohistochemistry (strept avidin-biotin complex, SABC method) and real-time quantitative PCR, respectively., Results: Compared with the normal group, the FVC, FEV0.1, FEV0.3, FEV0.1/FVC and FEV0.3/FVC levels were significantly decreased ( P <0.01), while the contents of TNF-α, TGF-α and IL-8 in the serum, BALF and lung tissues, expression levels of EGFR, p38MAPK and MUC5AC mRNAs and proteins, and the immunoactivity of EGFR, p38MAPK and MUC5AC in the lung tissues were significantly increased in the model group ( P <0.01). After EA intervention, the decreased levels of the FVC, FEV0.1, FEV0.3, FEV0.1/FVC and FEV0.3/FVC, and the increased levels of the abovementioned genes and proteins were all reversed in the EA group ( P <0.01, P <0.05). After modeling, the bronchial walls were thickened, with enlarged alveolar cavities, fractured alveolar walls, obvious inflammatory cell infiltration, and rich mucus secretion in the lumen, which was relatively milder in the EA group., Conclusion: EA of ST36 can improve the ventilation function in COPD rats, which may be associated with its function in down-regulating the levels of TNF-α, TGF-α, IL-8, EGFR, p38MAPK and MUC5AC mRNAs and proteins in the lung tissues, inhibiting EGFR-p38MAPK signaling mediated expression of MUC5AC.

In order to enrich the transcriptome data of Fagopyrum dibotrys plants, analyze the genes encoding key enzyme involved in flavonoid biosynthesis pathway, and mine their functional genes, in this study, we performed RNA sequencing analysis for the rhizomes, roots, flowers, leaves and stems of F. dibotrys on the BGISEQ-500 sequencing platform. After de novo assembly of transcripts, a total of 205 619 unigenes were generated and 132 372 unigenes were obtained and annotated into seven public databases, of which, 81 327 unigenes were mapped to the GO database and most of the unigenes were annotated in cellular process, biological regulation, binding and catalytic activity. Besides, 86 922 unigenes were enriched in 136 pathways using KEGG database' and we identified 82 unigenes that encodes key enzymes involved in flavonoid biosynthesis. Comparing rhizome with root, flower, leaf or stem in F. dibotrys, 27 962 co-expressed differentially expressed genes(DEGs) were obtained. Among them, 23 515 DEGs of rhizome tissue-specific were enriched into 132 pathways and 13 unigenes were significantly enriched in biosynthesis of flavone and flavonol. In addition, we also identified 3 427 unigenes encoding 60 transcription factor(TFs) families as well as four unigenes encoding bHLH TFs were enriched in flavonoid biosynthesis. Our results greatly enriched the transcriptome database of plants, provided a reference for the analysis of key enzymes involved in flavonoid biosynthesis in plants, and will facilitate the study of the functions and regulatory mechanisms of key enzymes involved in flavonoid biosynthesis in F. dibotrys at the genetic level.

Based on the original text of Huangdi Neijing ( Yellow Emperor's Internal Classic ), the theories of biaoben , genjie and qijie are analyzed deeply and the practical significance is explored on the correlation with meridians and zangfu organs. Biaoben , genjie and qijie represent respectively the relevant rules between the body surface and internal organs in different patterns and they are the foundation of the multiple correlations of the upper and the lower, the internal and the external and the horizontal and the transverse between meridians and zangfu organs. The understanding above not only contributes to the recognition and the exploration of the relevant essence of meridians and zangfu organs, but also provides the theoretic guidance to the acupoint selection, diagnosis and treatment of zangfu disorders in clinical practice.

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-1β, 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-1β, 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-1β, 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-1β, 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.

Objective: To observe the effect of moxibustion on serum inflammatory factors, Toll-like receptor 4 (TLR4), myeloid differentiation factor 88 (MyD88) and nuclear factor Kappa B (NF-κB) in colon tissue of rats with diarrhea-predominant irritable bowel syndrome (IBS-D)，so as to explore the mechanism of moxibustion in the treatment of IBS-D., Methods: Healthy male SD rats were randomly divided into blank group, model group and moxibustion group, with 8 rats in each group. The IBS-D model was established by chronic restraint combined with gavage of Senna-leaf solution. The rats of the moxibustion group was treated with moxibustion at "Tianshu"(ST25)and "Shangjuxu"(ST37)for 30 min， once a day for 7 d. After the intervention, the rate of loose stools and the minimum threshold volume of abdominal withdrawal reflex (AWR) induced by colorectal distension were observed. The contents of serum inflammatory factors IL-1β, TNF-α and IL-6 were detected by ELISA. The pathological changes of rat colon were observed by H.E. staining. The average optical density of TLR4 and NF-κB (p65) in colon tissue was detected by immunohistochemistry. The relative expressions of TLR4, MyD88 and NF-κB (p65) mRNAs in colon tissues were detected by real-time fluorescence quantitative PCR, and the expression levels of TLR4, MyD88 and NF-κB (p65) proteins in colon tissues were detected by Western blot., Results: There was a mild degree of inflammation in colon in rats of the model group. Compared with the blank group, the minimum volume threshold of AWR decreased significantly ( P < 0.01), and the rate of loose stools, the contents of IL-1β, IL-6 and TNF-α in serum as well as the expressions of TLR4, MyD88, NF-κB (p65) mRNAs and protein in colon tissues were significantly increased in the model group ( P <0.01). Compared with the model group, the inflammation was alleviated in colonic mucosa, the minimum volume threshold of AWR increased significantly ( P <0.01), and rate of loose stools decreased significantly ( P <0.01) in the moxibustion group, the contents of IL-1β, TNF-α and IL-6 in serum and the expression levels of TLR4, MyD88, NF-κB (p65) mRNAs and proteins in colon tissue decreased significantly in the moxibustion group ( P <0.01)., Conclusion: Moxibustion ST25 and ST37 can improve diarrhea symptom and visceral hypersensitivity in IBS-D rats, which may be related with its effects in inhibiting TLR4/MyD88/NF-κB signaling pathway and reducing the expression of inflammatory factors.

Objective: To observe the effect of electroacupuncture (EA) preconditioning on the expression of liver protein kinase 1 (LKB1), adenosine 5'-monophosphate (AMP)-activated protein kinase (AMPK) and 6-phosphofructo-2-kinase (PFK2) in cardiomyocytes of rats with acute myocardial ischemia (AMI), so as to explore its mechanisms underlying cardioprotective effect., Methods: Thirty male Wistar rats were randomly divided into sham-operation, model and EA pretreatment groups ( n ＝10 rats per group). The AMI model was established by ligation of the left anterior descending branch of coronary artery. Before modeling, EA preconditioning (2 Hz/15 Hz, 1 mA) was applied to bilateral "Neiguan"(PC6) for 30 min, once daily for 14 days. Histopathological changes of myocardium was observed by microscope after H．E. staining. The level of lactate dehydrogenase (LDH) in serum was detected by ELISA. The expression of autophagy-associated proteins and mRNAs as LKB1, AMPKa1, AMPKa2 and PFK2 were detected by Western blot and real-time PCR, respectively., Results: Compared with the sham-operation group, serum LDH content, and expression levels of myocardial AMPKa2 and PFK2 proteins and mRNAs were significantly up-regulated ( P <0.01), and those of LKB1 and AMPKa1 proteins and mRNAs were increased in the model group ( P <0.05). Following the intervention, serum LDH were apparently down-regulated ( P <0.01), and expression levels of myocardial LKB1, AMPKa1 and PFK2 proteins and mRNAs were apparently up-regulated ( P <0.01), but that of AMPKa2 protein and mRNA was remarkably down-regulated in the EA group ( P <0.01). H．E. staining showed cell swelling, disordered arrangement of myocardial fibers with obvious rupture, interstitial bleeding and inflammatory infiltration, which was relatively milder in the EA preconditioning group., Conclusion: EA pretreatment can trigger LKB1/AMPK/PFK2 signaling pathway in AMI rats, which may contribute to its cardioprotective effect against ischemic myocardial injury by activating autophagy of cardiomyocytes.　.

Objective: To explore the effect of moxibustion of acupoints of Governor Vessel on learning-memory ability and expression of the receptor for advanced glycation end products (RAGE), low density lipoprotein (LDL) receptor related protein-1 (LRP-1), β-amyloid precursor protein (APP) and neuronal ultrastructure of the frontal cortex and hippocampus in vascular dementia (VD) rats, so as to investigate its underlying mechanisms in relieving VD., Methods: A total of 24 male SD rats were randomized into normal, model, and moxibustion groups ( n ＝8 rats in each group). The VD model was established by occlusion of the bilateral common carotid arteries for 20 min and reperfusion for 10 min which was repeated for 3 times. Moxibustion was applied to "Baihui" (GV20), "Dazhui"(GV14) and "Fengfu"(GV16)for 30 min, once daily for 4 weeks. The learning-memory ability was evaluated by using Morris water maze tests. At the end of experiments, the frontal lobe of cerebral cortex and hippocampus tissues were collected for detecting the expression of RAGE and LRP-1 proteins with Western blot, and RAGE, LRP-1 and APP mRNAs with quantitative real-time PCR, respectively. The ultrastructure of neurons in the frontal cortex and hippocampal CA1 region was observed with transmission electron microscopy., Results: Following modeling, Morris water maze tests showed that the average escape latency of the model group and moxibustion group was significantly prolonged on the 4 th day of modeling ( P <0.01). The expression levels of RAGE protein and mRNA and APP mRNA were significantly increased ( P <0.01), while those of LRP-1 protein and mRNA in the cortex and hippocampus remarkably decreased in the model group relevant to the normal group ( P <0.01). Following moxibustion, modeling-induced increase of expression of RAGE protein and mRNA and APP mRNA, and down-regulation of expression of LRP-1 protein and mRNA were reversed in the moxibustion group relevant to the model group ( P <0.01). Results of electron microscopy showed dilation of the mitochondria with disappearance of the cristae, partial vacuolar degeneration or dissolved external membrane, and cytoplasmic edema with basic disappearance of the rough endoplasmic reticulum and glycogen particles in neurons of the frontal cortex and hippocampal CA1 region after modeling, which was relatively milder in the moxibustion group., Conclusion: Moxibustion of acupoints of the Governor Vessel may reduce the content of APP in the frontal cortex and hippocampus by regulating the expression of RAGE and LRP-1 proteins and mRNA in VD rats.

Objective: To observe the effect of acupuncture plus moxibustion on the synaptic ultrastructure and expression of synaptic skeleton related proteins in the prefrontal cortex (PFC) of heroin re-addicted rats, so as to reveal its mechanisms underlying improvement of heroin addiction., Methods: Twenty-four Wister rats (half male and half female) were randomly divi-ded into normal control, model and acupuncture groups ( n ＝8 in each group). The heroin re-addicted model was established by muscular injection of heroin into the hind limbs for 8 days (incremental 0.8-3.6 mg, once daily for 6 days, and twice daily for 2 days), followed by conventional breeding for 5 days (detoxification), the procedure (addition－detoxification) was repeated 3 cycles. For rats of the acupuncture group, "Baihui" (GV20) was needled with filiform needles which were retained for 30 min, and moxibustion was then applied to bilateral "Shenshu" (BL23) for 30 min. The treatment was conducted once daily during the deto-xification. On the 39 th day of experiment, the bilateral prefrontal cortex tissues were sampled for examining the ultrastructure by using transmission electron microscope (TEM) after fixative solution immersion and for determining the expression of genes and proteins of activity-regulated cytoskeleton-associated protein (Arc), microtubule-asso-ciated protein-2 (MAP-2) and microtubule-associated protein Tau (Tau) with quantitative real-time PCR and Western blot, respectively., Results: After modeling, the expression levels of Arc mRNA and protein were significantly up-regulated, and those of MAP-2 and Tau mRNA and proteins ob-viously down-regulated in the model group relevant to the normal control group ( P <0.05). Following the intervention, the up-regulated Arc protein and mRNA and the down-regulated MAP-2 and Tau were obviously reversed relevant to the model group ( P <0.05). Outcomes of TEM showed unclear pre- and post-membranes of the synapses, narrowing of the synaptic gap and non-uniform of the density of the thickened dense plaque after modeling, which was relatively milder in the acupuncture group., Conclusion: Acupuncture plus moxibustion can improve changes of synaptic ultrastructure in heroin re-addicted rats, which may be related to their effect in regulating the expression of some synaptic skeleton proteins and genes.

The hippocampus is involved in the regulation of the autonomic nervous system, together with the hypothalamus and brainstem nuclei, such as the paraventricular nucleus and nucleus tractus solitarius. The vagus nerve-nucleus tractus solitarius pathway has an important role in cardiovascular reflex regulation. Myocardial ischemia has been shown to cause changes in the autonomic nervous system, affecting the dynamic equilibrium of the sympathetic and vagal nerves. However, it remains poorly understood how the hippocampus communicates with brainstem nuclei to regulate the autonomic nervous system and alleviate myocardial ischemic tissue damage. A rat model of acute myocardial ischemia (AMI) was made by ligating the left anterior descending branch of the coronary artery. Three days before ischemia, the hippocampal CA1 region was damaged. Then, 3 days after ischemia, electroacupuncture (EA) at Shenmen (HT7)-Tongli (HT5) was performed (continuous wave, 1 mA, 2 Hz, duration of 30 minutes). Cluster analysis of firing patterns showed that one type of neuron was found in rats in the sham and AMI groups. Three types of neurons were observed in the AMI + EA group. Six types of neurons were found in the AMI + EA + Lesion group. Correlation analysis showed that the frequency of vagus nerve discharge in each group was negatively correlated with heart rate (HR) (P < 0.05, r = -0.424), and positively correlated with mean arterial pressure (MAP) (P < 0.05, r = 0.40987) and the rate-pressure product (RPP) (P < 0.05, r = 0.4252). The total frequency of the nucleus tractus solitarius discharge in each group was positively correlated with vagus nerve discharge (P < 0.01, r = 0.7021), but not with hemodynamic index (HR: P > 0.05, r = -0.03263; MAP: P > 0.05, r = -0.08993; RPP: P > 0.05, r = -0.03263). Some neurons (Neuron C) were negatively correlated with vagus nerve discharge, HR, MAP and RPP in the AMI + EA group (vagus nerve discharge: P < 0.05, r = -0.87749; HR: P < 0.01, r = -0.91902; MAP: P < 0.05, r = -0.85691; RPP: P < 0.01, r = -0.91902). Some neurons (Neurons C, D and E) were positively correlated with vagus nerve discharge, HR, MAP and RPP in the AMI + EA + Lesion group (vagus nerve discharge: P < 0.01, r = 0.8905, P < 0.01, r = 0.9725, P < 0.01, r = 0.9054; HR: P < 0.01, r = 0.9347, P < 0.01, r = 0.9089, P < 0.05, r = 0.8247; MAP: P < 0.05, r = 0.8474, P < 0.01, r = 0.9691, P < 0.01, r = 0.9027; RPP: P < 0.05, r = 0.8637, P < 0.01, r = 0.9407, P < 0.01, r = 0.9027). These findings show that the hippocampus-nucleus tractus solitarius-vagus nerve pathway is involved in the cardioprotective effect of EA at the heart meridian. Some interneurons in the nucleus tractus solitarius may play a particularly important role in the cardiomodulatory process., Competing Interests: All authors declare no personal or financial conflicts of interest

Objective: We have repeatedly demonstrated that electroacupuncture (EA) stimulation of "Shenmen" (HT 7)- "Tongli" (HT 5) segment of the Heart Meridian can improve acute myocardial ischemia (AMI). This study aimed at observing the effect of EA on contents of hippocampal norepinephrine (NE), and interleukin 6 (IL-6), IL-1 β and tumor necrosis factor- α (TNF-α) in AMI rats, so as to explore its underlying mechanism., Methods: SD rats were randomly divided into 3 groups: sham operation (sham), model and EA( n =6 rats in each). The anterior descending branch (ADB) of the left coronary artery was occluded to make an AMI model. For rats of the sham group, a surgical suture was simply threaded beneath the ADB without ligation. EA (2 Hz/15 Hz, 1 mA) was applied to bilateral "Shenmen" (HT 7)- "Tongli" (HT 5) and the middle-point between HT 7 and HT 5 for 30 min, once daily for 3 days. Electrocardiogram (ECG) of the neck-thoracic lead was recorded by using PowerLab 16. The contents of serum creatine kinase (CK), hippocampal IL-6, IL-1 β and TNF-α were assayed by ELISA. The concentration of NE in hippocampal CA 1 area was detected by microdialysis combined with electrochemical detector., Results: Compared with the sham group, the ECG-ST height, serum CK, hippocampal NE, IL-6, IL-1 β and TNF-α contents of the CA 1 region were significantly increased in the model group ( P <0.001). Whereas, after EA intervention, the serum CK, hippocampal NE, IL-6, IL-1 β and TNF-α contents were obviously down-regulated relevant to the model group ( P <0.05, P <0.001), and the IL-6, IL-1 β and TNF-α contents were positively correlated with the NE level ( P <0.001, P <0.01)., Conclusion: EA stimulation of the Heart Meridian may improve ischemic myocardial injury in AMI rats, probably by reducing the proinflammatory factors and hippocampal NE.

To explore the activity of essential oil extracted from Artemisia argyi (AAEO) in inducing the apoptosis of Candida albicans SC5314. The effect of AAEO on reactive oxygen species(ROS) and mitochondria membrane potential(MMP) of C. albicans SC5314 was detected by flow cytometry. Phosphatidylserine externalization was observed under fluorescence microscopic with Annexin-V/PI staining at the early stage of apoptosis in C. albicans. Metacaspase activity was observed under fluorescence microscopic with FITC-VAD-FMK staining at the early stage of apoptosis in C. albicans. C. albicans morphology was observed by DAPI nuclear staining and fluorescence microscopy. After intervention with 0.5 mL•L⁻¹ AAEO, apoptosis of C. albicans significantly increased, metacaspase activity increased, nuclear pyknosis and fragmentation, and intracellular ROS were significantly increased, and mitochondrial membrane potential decreased significantly. The certain concentrations of AAEO could induce the apoptosis of C. albicans., Competing Interests: The authors of this article and the planning committee members and staff have no relevant financial relationships with commercial interests to disclose., (Copyright© by the Chinese Pharmaceutical Association.)

Objective: To observe the effect of acupuncture serum on the number of apoptosis of the cultured hippocampal neurons with seizure-like discharges and the expression levels of endoplasmic reticulum stress-inducible molecular chaperones glucose-regulated protein 78 (GRP 78), C/EBP homologous protein (CHOP) and cysteine protease protein-12(Caspase-12), so as to reveal its protective mechanism on seizure-induced injury of hippocampal neurons., Methods: A regular primary culture of neurons derived from the hippocampus of the newly-born SD rats was conducted for 10 days, then these cultured neurons that displayed seizure-like discharges in Mg 2+ -free medium were divided into normal extracellular fluid (medium) group (normal), Mg 2+ -free medium group, acupuncture serum group and non-acupuncture serum group ( n =30). Blood examples were taken from pentylenetetrazol (i.p.i.) -induced acute convulsion adult SD rats underwent manual acupuncture stimulation of "Baihui" (GV 20) and "Dazhui" (GV 14, once daily for 7 days) to prepare acupuncture serum. Hippocampal neuronal cultures were prepared from hip-pocampal tissue isolated from 24 h-old SD rats. The isolated neurons were incubated normally in Dulbecco's Modified Eagle Me-dium (DMEM)/Nutrient Mixture F 12(1:1) for 10 days, followed by exposure to the extracellular fluid {composed of NaCl, KCl, CaCl 2 , MgCl 2 , HEPES[4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid], D-glucose and aminoethanoic acid (pH 7.3)} for 3 h and returned to normal culture for normal group, by exposure to Mg 2+ -free extracellular fluid (to induce seizure-like discharges) for 3 h and returned to normal culture for Mg 2+ -free medium group, by exposure to Mg 2+ -free extracellular fluid for 3 h and returned to a regular culture medium[DMEM/F 12(1:1)] plus acupuncture serum (9:1) for acupuncture serum group, and by exposure to Mg 2+ -free extracellular fluid for 3 h and returned to DMEM/F 12(1:1) plus non-acupuncture serum (9:1) for non-acupuncture serum group. The cell apoptosis was assessed at 2, 12 and 48 h after application of acupuncture serum or non-acupuncture serum by using TUNEL method and the expression levels of GRP 78, CHOP and Caspase-12 proteins of the cultured cells at the 3 time-points detected using Western blot., Results: The number of apoptotic hippocampal neurons was significantly higher in the Mg 2+ -free medium group than in the normal medium group at 2, 12 and 48 h ( P <0.01), and considerably decreased in the acupuncture serum group (but not in the non-acupuncture serum group) relevant to the Mg 2+ -free medium group at the same time-points after application of acupuncture serum ( P <0.05, P <0.01). The expression levels of GRP 78 protein at 2 h, CHOP and Caspase-12 proteins at 2, 12 and 48 h were significantly up-regulated in the Mg 2+ -free medium group relavant to the normal me-dium group ( P <0.05, P <0.01). After application of acupuncture serum to the culture medium, the expression levels of GRP 78 protein at the three time-points were significantly increased in comparison with the Mg 2 + -free medium group ( P <0.01, P <0.05), while those of CHOP at 12 and 48 h, and Caspase-12 at the three time-points were notably down-regulated ( P <0.05, P <0.01). No significant differences were found between the non-acupuncture serum and Mg 2 + -free medium groups in the expression levels of GRP 78, CHOP and Caspase-12 proteins at the three time-points ( P >0.05)., Conclusions: Acupuncture serum can significantly reduce apoptosis of the cultured hippocampal neurons, which may be related to its effects in increasing the expression of GRP 78 protein and down-regulating the expression of CHOP and Caspase-12 proteins, suggesting an important role of acupuncture serum in maintaining the stability of the neuronal endoplasmic reticulum.

Objective: To observe the effect of acupuncture stimulation of different acupoints or acupoint groups on intestinal locomotor in slow transit constipation (STC) mice, so as to provide experimental evidence for demonstrating the synergic function of acupoint combination., Methods: A total of 190 Kunming mice were randomized into 19 groups:normal control, model, uni-"Quchi" (LI 11), uni-"Shangjuxu" (ST 37), uni-"Tianshu"(ST 25), and uni-"Dachangshu"(BL 25) groups, for analyzing the effect of single acupoint; normal control, model, uni-ST 25+uni-BL 25, uni-LI 11+uni-ST 37, bi-ST 25, bi-BL 25, bi-LI 11, and bi-ST 37 groups, for comparing the effect between single acupoint and two acupoints combination; normal control, model, bi-ST 25+bi-BL 25, bi-LI 11+bi-ST 37, and uni-LI 11+uni-ST 37+uni-ST 25 +uni-BL 25 groups, for comparing the effect between four acupoints and two acupoints combinations (10 mice/group). The STC model was established by intragastric administration of compound diphenoxylate suspension (1 mg/100 g), once daily for 2 days. Manual acupuncture needle stimulation (200 twirling times/min for 1 min) was applied to the abovementioned unilateral or bilateral acupoints twice daily for 7 times. The first defecation time, the defecation grain number in 12 h and the intestinal propulsion rate were determined using activated carbon gavage., Results: Compared with the normal group, the first defecation time was significantly increased in the STC model group ( P <0.01), while the 12 h defecation grain number and the intestinal propulsion rate were considerably decreased in the model group ( P <0.01). After manual acupuncture intervention, the first defecation time was notably shortened in the uni-LI 11, uni-ST 37, uni-ST 25, uni-BL 25, bi-ST 25, bi-BL 25, bi-LI 11, bi-ST 37, uni-ST 25+uni-BL 25, uni-LI 11+uni-ST 37, bi-ST 25+bi-BL 25, bi-LI 11+bi-ST 37 and the uni-ST 25+uni-BL 25+uni-LI 11+uni-ST 37 groups compared with the model group ( P <0.05), while both the 12 h defecation grain number and intestinal propulsion rate were considerably increased in the abovementioned 13 acupuncture groups ( P <0.05, P <0.01). The effects of uni-ST 25+uni-BL 25 and uni-LI 11+uni-ST 37 were significantly superior to those of bi-ST 25, bi-BL 25, bi-LI 11 and bi-ST 37 groups, and those of the uni-ST 25+uni-BL 25+uni-LI 11+uni-ST 37 group were markedly superior to those of bi-ST 25+bi-BL 25 and bi-LI 11+bi-ST 37 groups in shortening the first defecation time and in increasing the 12 h defecation grain number and intestinal propulsion rate ( P <0.05)., Conclusions: Acupuncture stimulation of LI 11, ST 37, ST 25 and BL 25 can improve constipation in STC mice, and the effect of two acupoints combination is significantly superior to that of the corresponding bilateral single acupoint, and that of the 4 unilateral single acupoint combination is obviously superior to that of two bilateral acupoints combination, suggesting a synergistic action of acupuncture stimulation.

Objective: To observe the effect of electroacupuncture (EA) stimulation of the Heart Meridian on electrical activities of neck sympathetic and vagal nerves in acute myocardial ischemia (AMI) rats., Methods: Thirty SD rats were randomly divided into 3 groups, sham operation (sham), model and EA( n =10 rats in each group). The left anterior descending branch (LADB) of the coronary artery was occluded to make an acute myocardial ischemia (AMI) model.For rats of the sham group, a surgical suture was simply threaded beneath the LADB without ligation. EA stimulation (2 Hz, 1 mA)was applied to bilateral "Shenmen" (HT 7), "Tongli" (HT 5) and the middle-point between HT 7 and HT 5 for 20 min. The electrical activities of the left cervical sympathetic and vagal nerves were recorded using a bipolar platinum wire electrodes and a BIOPAC MPl 50 system, and the electrocardiogram (ECG) of the standard limb lead Ⅱ was recorded simultaneously. The AMI severity was evaluated by using J-spot height of ECG., Results: After AMI, the heart rate (HR), J-spot of ECG, and the discharge frequency of the cervical sympathetic nerve were significantly increased in the model group compared with the sham group ( P <0.01), while the discharge frequency of vagal nerve was obviously decreased ( P <0.01). Following EA stimulation, the increased levels of HR, J-spot height and discharge frequency of sympathetic nerve, and the decreased discharge frequency of vagus were significantly suppressed at time-points of 1, 3, 5 and 15 min after EA ( P <0.01). In addition, the ratio of discharge frequencies of vagal/sympathetic nerves was notably down-regulated after AMI ( P <0.01). After EA stimulation, the ratios of vagal/sympathetic discharge at the time-points of 1, 3, 5 and 15 min following EA were significantly increased ( P <0.01)., Conclusions: EA stimulation of the Heart Meridian may improve AMI by inhibiting sympathetic discharges and elevating vagal discharges in AMI rats, i.e., coordinating activities of the autonomic nerve systems.

Objective: To observe the effect of electroacupuncture (EA) of "Dazhui" (GV 14), "Baihui" (GV 20), etc. on learning-memory ability and expression of vascular endothelial growth factor (VEGF) mRNA, VEGF receptor 1 (VEGFR-1/Flt-1) mRNA, and VEGFR-2 (Flk-1) mRNA in the hippocampus in vascular cognitive impairment (VCI) rats so as to reveal its mechanism underlying improvement of VCI., Methods: A total of 60 Wistar rats were randomly divided into sham operation, VCI model, EA and medication groups (n=12 in each group). The VCI model was established by occlusion of the bilateral vertebral arteries and bilateral cervical arteries. EA (2 Hz/20 Hz) was applied to "Baihui" (GV 20), "Dazhui" (GV 14), "Shuigou" (GV 26) and "Shenting" (GV 24) for 20 min, once daily for 20 days. Rats of the medication group were treated by intragastric perfusion of Aniracetam capsules (0.0625 g/kg),once daily for 20 days. The rats' learning-memory ability was detected by step-down test. The expression levels of VEGF mRNA, Flt-1 mRNA and Flk-1 mRNA in the hippocampus were detected by RT-PCR, and the neurological deficit scores were assessed by Zea Longa (0-4 scaling) method., Results: Compared with the sham operation group, rats of the model group showed a significant increase in reaction time and error number and decrease of escape latency (reduction of learning-memory ability), and increase in neurological deficit score, and in expression levels of hippocampal VEGF mRNA, Flt-1 mRNA and Flk-1 mRNA (all P<0.01). After EA treatment, in comparison with the model group, the learning-memory ability and hippocampal VEGF mRNA, Flt-1 mRNA and Flk-1 mRNA were apparently increased, neurological deficit score were markedly decreased in the EA group (all P<0.01). The effects of EA treatment was obviously superior to those of medication in raising learning-memory ability and up-regulating hippocampal VEGF mRNA and Flt-1 mRNA expression levels, and in reducing neurological deficit score (all P<0.05)., Conclusion: EA intervention can apparently up-regulate hippocampal VEGF mRNA, Flt-1 mRNA and Flk-1 mRNA expression in VCI rats, which may contribute to its effect in improving learning-memory ability, possibly by promoting neovasculization.

Five hundred and six fresh fecal samples were collected from Lu'an, Fuyang, Suzhou, Chizhou, Wuhu, Chuzhou and Bozhou in Anhui Province, and detected firstly by direct smear microscopy. The microscopy-positive samples were amplified by nested PCR targeting the triosephosphate isomerase (TPI) and glutamate dehydrogenase(GDH) genes. The positive PCR products were sequenced in both directions. The sequences were analyzed by ClustalX 1.81 for sequence alignment and the neighbor-joining trees were constructed by Mega 5.05. Thirty-two out of 506 fecal specimens were diagnosed as Giardia-positive by microscopy with an infection rate of 6.3%. 23 and 16 of the samples were typed as assemblage E by the TPI (530 bp) and GDH (450 bp) genes, respectively. These findings indicated that there was a different distribution of subtypes of assemblage E in different areas. The zoonosis genotypes such as assemblage A or B was not found in the present study.