[Electroacupuncture at "Baihui"（GV20） and "Yongquan"（KI1） improves learning-memory ability of APP/PS1 double transgenic mice by regulating endosomal-lysosomal system].

OBJECTIVE: To explore the mechanism of electroacupuncture（EA） in improving learning-memory ability in Alzheimer's disease （AD） mice from the perspective of endosomal-lysosomal system. METHODS: Male APP/PS1 transgenic mice were randomly divided into model group and EA group （n=10 in each group） and 10 male C57BL/6 wild mice were taken as the normal group. EA （1 Hz/50 Hz, 1 mA） was applied at bilateral "Yongquan"（KI1） and acupuncture was applied at "Baihui" （GV20） for 15 min. The mice of the model and normal groups were subjected to restriction with the same method as those of the EA group for 15 min. The treatment was conducted once every other day for 6 weeks. The spatial learning-memory ability （shown by escape latency of place navigation test and the time of crossing the target platform and total swimming distance in the target quadrant in 1 min of spatial probe test ） was detected by Morris water maze test. The immunoactivity of senile plaques （SP） in the hippocampus tissue was detected by immunohistochemistry. The ultrastructural characters of hippocampal neurons were observed by transmission electron microscope, and the expression levels of Ras-related protein 5 （Rab5）, Ras-related protein 7 （Rab7） and cathepsin D （CTSD） in the hippocampus were detected by Western blot, separately. RESULTS: Compared with the normal group, the escape latency, SP immunoactivity, and protein expression levels of Rab5, Rab7 and CTSD were significantly increased （P<0.05, P<0.01）, while the number of crossing the original platform and the total swimming distance in the platform quadrant were considerably reduced （P<0.05） in the model group. In contrast to the model group, the EA group had a marked decrease in the escape latency, SP immunoactivity, and protein expression levels of Rab5, Rab7 and CTSD （P<0.05, P<0.01）, and a striking increase in the number of crossing the original platform and the swimming distance in the platform quadrant （P<0.05）. Results of transmission electron microscope showed an accumulation of endosome, lysosome, and endolysosomes in the hippocampal neurons in the model group, which was evidently milder in the EA group. CONCLUSION: EA of GV20 and KI1 can improve the learning-memory ability of AD mice, which may be related to its function in reducing hippocampal Aß deposition and down-regulating endosomal-lysosomal system activity.

[Effect of electroacupuncture on liver Akt/FoxO1 signaling pathway in rats with diabetic fatty].

OBJECTIVE: To observe the effect of electroacupuncture (EA) on liver protein kinase B (Akt)/forkhead box transcription factor 1 (FoxO1) signaling pathway in Zucker diabetic fatty (ZDF) rats, and to explore the possible mechanism of EA on improving liver insulin resistance of type 2 diabetes mellitus. METHODS: Twelve male 2-month-old ZDF rats were fed with high-fat diet for 4 weeks to establish diabetes model. After modeling, the rats were randomly divided into a model group and an EA group, with 6 rats in each group. In addition, six male Zucker lean (ZL) rats were used as the blank group. The rats in the EA group were treated with EA at bilateral "Zusanli" (ST 36), "Sanyinjiao" (SP 6), "Weiwanxiashu" (EX-B 3), and "Pishu" (BL 20). The ipsilateral "Zusanli" (ST 36) and "Weiwanxiashu" (EX-B 3) were connected to EA device, continuous wave, frequency of 15 Hz, 20 min each time, once a day, six times a week, for a total of 4 weeks. The fasting blood glucose (FBG) in each group was compared before modeling, before intervention and after intervention; the serum levels of insulin (INS) and C-peptide were measured by radioimmunoassay method, and the insulin resistance index (HOMA-IR) was calculated; HE staining method was used to observe the liver tissue morphology; Western blot method was used to detect the protein expression of Akt, FoxO1 and phosphoenolpyruvate carboxykinase (PEPCK) in the liver. RESULTS: Before intervention, compared with the blank group, FBG was increased in the model group and the EA group (P<0.01); after intervention, compared with the model group, FBG in the EA group was decreased (P<0.01). Compared with the blank group, the serum levels of INS and C-peptide, HOMA-IR, and the protein expression of hepatic FoxO1 and PEPCK were increased (P<0.01), while the protein expression of hepatic Akt was decreased (P<0.01) in the model group. Compared with the model group, the serum levels of INS and C-peptide, HOMA-IR, and the protein expression of hepatic FoxO1 and PEPCK were decreased (P<0.01), while the protein expression of hepatic Akt was increased (P<0.01) in the EA group. In the model group, the hepatocytes were structurally disordered and randomly arranged, with a large number of lipid vacuoles in the cytoplasm. In the EA group, the morphology of hepatocytes tended to be normal and lipid vacuoles were decreased. CONCLUSION: EA could reduce FBG and HOMA-IR in ZDF rats, improve liver insulin resistance, which may be related to regulating Akt/FoxO1 signaling pathway.

[Perpendicular and transverse needling of "Sanyinjiao" (SP6) relieves abdominal pain by regulating arginine vasopressin and its receptor expression levels in uterus and hypothalamus in cold-stasis type dysmenorrhea rats].

OBJECTIVE: To observe the effect of perpendicular and subcutaneous transverse needling at "Sanyinjiao" (SP6) on visceral pain behavior, arginine vasopressin (AVP) content in the serum, uterine tissues, spinal cord and hypothalamus and expression of AVP receptors AVPR1A and AVPR1B in the uterine tissues, spinal cord and hypothalamus in cold-stasis (stasis due to pathogenic cold) type dysmenorrhea rats, so as to explore their mechanisms underlying pain relief. METHODS: Forty female SD rats were randomly divided into blank control, model, perpendicular needling and transverse needling groups, with 10 rats in each group. The cold-stasis dysmenorrhea rat model was established by exposure in a freezer (-25 ℃) for 4 h, once daily for 5 days, and subcutaneous injection of estradiol benzoate (once daily for 10 days) and intra-abdominal injection of oxytocin injection (once). For rats of the two acupuncture groups, acupuncture needles were inserted into the bilateral SP6 perpendicularly or transversely to a depth of about 4-5 mm, and retained for 20 min. The abdominal pain behavior was assessed by recording the writhing latency and scaling the rats' writhing reactions after modeling. The contents of AVP in the serum, uterus, spinal cord and hypothalamus tissues were assayed using ELISA and the expression of AVPR1A and AVPR1B in the uterus, spinal cord and hypothalamus was measured by using Western blot and quantitative real time-PCR, respectively. RESULTS: After mode-ling and compared with the blank control group, the writhing latency was significantly shortened (P<0.05), and the writhing score in the first 20 min was significantly increased (P<0.01) in the model group. After the intervention, the writhing latency was significantly prolonged (P<0.01), and the writhing scores in 20 min were significantly decreased (P<0.01) in the two needling groups. The AVP contents were obviously increased in the serum and uterine tissue (P<0.05, P<0.01) but decreased appa-rently in the spinal cord and hypothalamus tissues (P<0.01, P<0.05), and the expression levels of AVPR1A or AVPR1B protein and mRNA were markedly increased in the uterine tissues (P<0.01, P<0.05), and significantly decreased in the spinal cord and hypotha-lamus in the model group relevant to the control group (P<0.05, P<0.01). Following the intervention, The AVP content in the serum of the perpendicular needling group (P<0.05) and that in the uterus of the two needling groups were significantly decreased (P<0.01), as well as that in the hypothalamus was obviously increased in the two needling groups (P<0.05). The expression levels of AVPR1A protein and mRNA in the uterus were significantly down-regulated in the two needling groups (P<0.01, P<0.05) and AVPR1B protein in the hypothalamus of the perpendicular needling group was up-regulated (P<0.05). Moreover, no significant differences were found between the two needling groups in regulating the related indexes mentioned above (P>0.05). CONCLUSION: Both perpendicular and subcutaneous transverse needling at SP6 have an immediate analgesic effect in cold-stasis type dysmenorrhea rats, which may be related to their effects in regulating AVP levels and its receptor expression in the uterine and hypothalamus.

Z-Shaped Transfer Functions for Binary Particle Swarm Optimization Algorithm.

Particle swarm optimization (PSO) algorithm is a swarm intelligent searching algorithm based on population that simulates the social behavior of birds, bees, or fish groups. The discrete binary particle swarm optimization (BPSO) algorithm maps the continuous search space to a binary space through a new transfer function, and the update process is designed to switch the position of the particles between 0 and 1 in the binary search space. Aiming at the existed BPSO algorithms which are easy to fall into the local optimum, a new Z-shaped probability transfer function is proposed to map the continuous search space to a binary space. By adopting nine typical benchmark functions, the proposed Z-probability transfer function and the V-shaped and S-shaped transfer functions are used to carry out the performance simulation experiments. The results show that the proposed Z-shaped probability transfer function improves the convergence speed and optimization accuracy of the BPSO algorithm.

Waggle needling wields preferable neuroprotective and anti-spastic effects on post-stroke spasticity rats by attenuating γ-aminobutyric acid transaminase and enhancing γ-aminobutyric acid.

Waggle needling, a classical anti-spastic needling technique characterized by combination of acupuncture with joint movement, has gained increasing popularity of spasticity treatment in China. This study was designed to compare the anti-spastic effect of waggle needling to the routine needling and to explore its underlying mechanism. We established post-stroke spasticity model based on ischemia stroke operation (middle cerebral artery occlusion). Rats were divided into six groups: normal control group, sham-operated control group, ischemia stroke model group, waggle needling group, routine needling group and baclofen group. Neurological function and muscle tone were assessed by the Zea Longa score and modified Ashworth scale, respectively. Indirect muscle tone was testified with electrophysiological recording. Cerebral infarction was measured by 2,3,5-triphenyltetrazolium chloride staining. The concentrations and expressions of γ-aminobutyric acid transaminase (GABAT) and γ-aminobutyric acid (GABA) were detected by enzyme-linked immunosorbent assay and western blot assay. Waggle needling markedly alleviated neurological deficits, decreased cerebral infarction and eased muscle tone; simultaneously, attenuated GABAT and enhanced GABA expression in the cortical infarct regions in comparison with the routine needling (P < 0.01), yet showed similar therapeutic effect to the baclofen group (P > 0.05). These results preliminary supported that waggle needling as a potential promising non-pharmacological intervention for the treatment of cerebral ischemia and spasticity.

[Effect of electroacupuncture of "Hegu" (LI4) and "Zusanli" (ST36) on intestinal sensitivity and motility in irritable bowel syndrome rats].

OBJECTIVE: To observe the effect of electroacupuncture (EA) of "Hegu"(LI4) and "Zusanli"(ST36)on changes of intestinal sensitivity and colonic motility and expression of colonic 5-hydroxytryptamine 3A receptor (5-HT3AR) in irritable bowel syndrome (IBS) rats, so as to reveal its mechanism underlying improvement of IBS. METHODS: A total of 40 neonatal Wistar rats were randomly and equally divided into normal control, model, LI4 and ST36 groups (n＝10). The IBS model was induced by mother-infant separation, acetic acid enema and colorectal distension (CRD). EA (2 Hz/100 Hz, a tolerable strength) was applied to bilateral LI4 and ST36 for 20 min, once every other day for 5 times. The Bristol stool form scale was used to assess the gastrointestinal function, and the latency and number of abdominal muscular contraction waves of abdominal withdrawal reflex (AWR) were used to evaluate the intestinal sensitivity and motility respectively. The immunoactivity of 5-HT3AR of the colon tissue was detected by immunohistochemistry. RESULTS: After modeling, the score of Bristol fecal form scale, number of muscular contraction waves and expression levels of colonic 5-HT3AR in the myometrium and mucosal layers were significantly increased (P<0.01), and the latency of muscular initial contraction wave was obviously shortened in the model group relevant to the normal control group (P<0.01). After the intervention, the increased Bristol fecal form score, number of muscular contraction waves and expression levels of 5-HT3AR in the myometrium and mucosal layers as well as the decreased latency of muscular contraction were reversed in both LI4 and ST36 groups (P<0.01, P<0.05). The effect of EA of ST36 was significantly superior to those of EA-LI4 in lowering Bristol fecal scale score and 5-HT3AR expression in the muscular layer (P<0.01), but obviously inferior to those of EA-LI4 in increasing the latency of of muscular initial contraction wave and down-regulating muscular contraction waves and 5-HT3AR expression in the mucosal layer (P<0.05, P<0.01). CONCLUSION: Both EA-LI4 and EA-ST36 can significantly improve the symptoms of abdominal pain and diarrhea, but EA-LI4 is better in suppressing intestinal high sensitivity, and EA-ST36 is better in promoting intestinal motility, suggesting a specificity of effect of acupoints of different meridians.

[Effect of electroacupuncture at "Zusanli" (ST 36) and "Yanglingquan" (GB 34) on perinatal nicotine-exposure-induced lung function and morphology of neonatal rats].

OBJECTIVE: To compare the effects of electroacupuncture (EA) at "Zusanli" (ST 36) versus "Yanglingquan" (GB 34) in the pregnant rats on perinatal nicotine-exposure-induced lung function and morphology of newborn rats and explore the rule of acupoint effect in EA for the prevention from lung dysplasia in newborn rats. METHODS: A total of 24 female SD rats were randomized into a normal saline group (S group), a nicotine group (N group), a nicotine-ST 36 group (N + ST 36 group) and a nicotine-GB 34 group (N+GB 34 group), 6 rats in each one. Starting at the 6th day of pregnancy, 0.9% sodium chloride solution was injected subcutaneously in the S group, 1 mg/kg; and in the rest 3 groups, nicotine of the same dose was injected through to the 21st postnatal day to establish the perinatal nicotine-exposure model. Simultaneously, during model preparation, EA was applied at "Zusanli" (ST 36) and "Yanglingquan" (GB 34) in the N+ST 36 group and the N+GB 34 group respectively, once a day, through to the 21st postnatal day. The lung function analytic system for small animal was adopted to observe the changes in lung function indicators in newborn rats, such as peak inspiratory flow (PIF), peak expiratory flow (PEF), expiratory resistance (RE), inspiratory resistance (RI) and dynamic compliance (Cdyn). HE staining was used to observe the morphological changes of lung, such as alveolar fusion and rupture. RESULTS: Compared with the S group, PEF and Cdyn were lower and PIF, RI and RE higher in the N group (all P<0.01), additionally, alveoli were fused and ruptured, alveolar wall thickened, the numbers of alveoli reduced, the interspace of alveoli enlarged and the diameter increased (P<0.01). Compared with the N group, in the N+ST 36 group, PEF and Cdyn were increased, PIF, RI and RE reduced (P<0.05, P<0.01), the alveolar fusion and rupture relieved, the numbers of alveoli increased, alveolar wall thinner, the interpsace of alveoli became normal and the diameter was reduced significantly (P<0.01). In the N+GB 34 group, the changes of lung function and morphological indicators were not significant (P>0.05). CONCLUSION: Electroacupuncture at "Zusanli" (ST 36) in the pregnant rats significantly improves the perinatal nicotine-exposure-induced lung function and morphology of newborn rats than electroacupuncture at "Yanglingquan" (GB 34).

[Comparison of effects of electroacupuncture at "Tianshu" (ST25) and "Dachangshu" (BL25) on intestinal sensitivity and expression of muscarinic M3R and 5-HT3AR in irritable bowel syndrome rats].

OBJECTIVE: To observe and compare the effect of electroacupuncture (EA) of "Tianshu" (ST25) and "Dachangshu" (BL25) on intestinal sensitivity and the expression of M3 muscarinic acetylcholine receptor (M3R) and 5-hydroxytryptamine 3A receptor (5-HT3AR) in the colon tissue in irritable bowel syndrome (IBS) rats, so as to explore its mechanism underlying improvement of IBS. METHODS: Forty newborn Wistar rats were randomly divided into blank control, model, ST25 and BL25 groups (n＝10 rats per group). The IBS model was established by joint application of maternal separation, acetic acid enema plus colorectal distension from day 8 to 21 after birth. At the age of 9 weeks, EA (2 Hz/100 Hz, 0.1－0.3 mA) was applied to ST25 and BL25 for 20 min, once every other day for 5 times. The modified method was used to record the abdominal withdrawal reflex (AWR), so as to evaluate the visceral sensitivity by referring to the modified Al-Chaer's and colleagues' methods. The threshold (initial wave), the number and peak-peak value of contraction waves (AWR) within 90 s were recorded. The immunoactivity of M3R and 5-HT3AR in the colon tissue was detected by immunohistochemistry. RESULTS: After modeling, the latency was obviously shortened, the number and peak-peak values of contraction waves of intestine were significantly increased, and the immunoactivity of M3R and 5-HT3AR was notably up-regulated in the model group compared with the blank control group (P<0.01). After the EA intervention, the latency was obviously prolonged, the peak-peak value in both ST25 and BL25 groups, and the number of contraction waves in the ST25 group were significantly decreased (P<0.01), and the expression levels of M3 and 5-HT3A R were considerably down-regulated in both ST25 and BL25 groups relevant to the model group (P<0.01). The therapeutic effects were significantly better in the ST25 group than in the BL25 group in prolonging the latency and reducing the contraction wave number, and in down-regulating the expression of colonic M3R and 5-HT3AR (P<0.05, P<0.01). CONCLUSION: EA at ST25 and BL25 can reduce the intestinal sensitivity in IBS rats, which is probably associated with its effects in down-regulating the expression of M3R and 5-HT3AR in the colon. The therapeutic effects of ST25 were obviously better than those of BL25 in suppressing intestinal sensitivity and expression of colonic M3R and 5-HT3AR.

[Electroacupuncture at "Zusanli" (ST36) and "Chize" (LU5) of mother rats exposed to nicotine during pregnancy and lactation has a protective effect on development of lung function and morphology in neonatal rats].

OBJECTIVE: To compare the different effect of electroacupuncture (EA) at "Zusanli" (ST36) and "Chize" (LU5) of mother rats exposed to Nicotine during pregnancy and lactation on lung function and morphological changes in offspring rats, so as to explore the most effective acupoint for improving the development of lung in neonatal rats. METHODS: A total of 24 female pregnancy SD rats were randomly divided into 4 groups: normal control, model, EA-ST36 and EA-LU5 (n＝6 rats in each group). Rats of the normal group were treated by subcutaneous injection of normal saline, and those of the other 3 groups treated by subcutaneous injection of nicotine (1 mg•kg－1•d－1) beginning from the 6th day to about the 21st day of pregnancy (childbirth day) for nicotine exposure during pregnancy and lactation. The daily EA treatment (2 Hz /15 Hz，1 mA) was applied to bilateral ST36 and LU5 for 20 min, beginning from the 6th day of pregnancy to the 21st day (childbirth day). The lung function of the offspring rats including the peak inspiratory flow (PIF), peak expiratory flow (PEF), lung resistance (RL), exhalation resistance (RE)and lung dynamic compliance (Cdyn) was detected by using a lung function analysis system. Histopathological changes (severity of alveolarization) of the offspring rats' lung tissue were observed under microscope after H．E. stain. RESULTS: Compared with the normal group, the PIF, RL and RE values were significantly increased (P<0.01), and PEF and Cdyn values significantly decreased in the model group (P<0.01). The alveolar diameter in the model group was evidently increased relevant to the normal group (P<0.01). Following the intervention, modeling induced increase of PIF, RL, RE and alveolar diameter and decrease of PEF and Cdyn values in the EA-ST36 group, and the increased PIF, RL and RE levels in the EA-LU5 group were obviously suppressed relevant to the model group (P<0.01, P<0.05). Additionally, modeling induced obvious congestion and edema of the alveolar wall, alveolar deformation, rupture and fusion, and reduction of the number of the pulmonary alveoli were evidently milder in both EA-ST36 and EA-LU5 groups. No significant differences were found between the EA-ST36 and EA-LU5 groups in the abovementioned 5 indexes of pulmonary function and alveolar diameter (P>0.05)．. CONCLUSION: EA of ST36 and LU5 of mother rats experiencing nicotine exposure during pregnancy and lactation can improve the lung function and morphological changes in neonatal rats, and the effect of ST36 is relatively better.

Effect of Electroacupuncture at Different Acupoints on the Expression of NMDA Receptors in ACC and Colon in IBS Rats.

OBJECTIVE: To observe the effects of electroacupuncture (EA) at different acupoints on the expression of N-methyl-D-aspartate receptor (NMDA receptor ) and behaviors in irritable bowel syndrome (IBS) rats. METHODS: Wistar rats were randomly divided into blank control group (blank group, n=10) and model preparation group (n=50); experimental rat model of IBS was established by the "neonatal maternal separation and acetic acid enema" combined with "colorectal distension stimulation" method. A total of 50 IBS rats were randomly assigned to five groups of 10 each: model group, Yintang (GV29) group, Neiguan (PC6) group, Tianshu (ST25) group, and Zusanli (ST36) group. Rats in four treatment groups, aged 9 weeks old, were treated with EA by HANS with a sparse-dense wave with a frequency of 2/100 Hz, current of 0.1-0.3mA, and 20 min/stimulation, every other day for a total of 5 sessions. After treatment, the abdominal visceral sensitivity was evaluated by abdominal withdrawal reflex (AWR), and the psychological and emotional behavior of rats were evaluated by the open-field test (OFT). The expression of NMDA receptors in anterior cingulate cortex (ACC) was detected by Quantitative Real-time PCR, and the positive expression of NMDA receptors in colon was detected by immunohistochemistry. RESULTS: The IBS rat's abdomen is more sensitive and irritable; NR1, NR2A, and NR2B in ACC and NR1 and NR2B in colon of rats significantly increased in the model group versus the normal group (P<0.01) and were inhibited in all treatment groups (P<0.01, P<0.05). Additionally, NR2A and NR2B in ACC reduced more in GV29 group (P<0.01) than in other treatment groups (P all<0.05) compared with the model group. The expression of NR2B in colon was significantly inhibited in ST36 group (P<0.01) and inhibited in GV29 group and ST25 group (P all <0.05) compared with the model group. And the expression of NR2B in colon was more inhibited in ST36 group than in PC6 group (P<0.01). CONCLUSIONS: EA at different acupoints could obviously relieve abdominal pain and abnormal behaviors in IBS rats in different degrees of effects. The effect of abdominal pain-relief, from greatest to least, is ST25, ST36, GV29, and PC6, while the effect of relieving abnormal behaviors caused by IBS, from greatest to least, is GV29, PC6, ST36, and ST25. There are significant differences in the expressions of NMDA receptors in ACC and colon among different acupoints. This difference should be related to the location distribution and indications of acupoints.

[Effect of Electroacupuncture at "Neiguan" (PC 6) and "Tianshu" (ST 25) for Colonic Motility and D 2 Receptor in Irritable Bowel Syndrome Rats].

OBJECTIVE: To observe and compare the effects of electroacupuncture (EA) at "Tianshu" (ST 25) and "Neiguan" (PC 6) for colonic motility and the expression of colon dopamine D 2 in irritable bowel syndrome (IBS) rats, and to explore the specificity of different meridians and different acupoints. METHODS: Forty Wistar newborn rats were randomly divided into blank, model, Tianshu and Neiguan groups. Separation of mother and child and acetic acid coloclyster combined with colorectal distension were used to establish IBS model in the model, Tianshu and Neiguan groups. At the age of 9 weeks, EA at bilateral ST 25 and PC 6 were applied in the corresponding groups 5 times, once every other day. After the intervention, the Bristol fecal score, the latent period of abdominal retraction reflex and the number of contraction waves were recorded. The expression of dopamine D 2 receptor was detected by immunohistochemistry. RESULTS: Compared with the blank group, the Bristol fecal score of the model group was higher (P<0.01), the 1st contraction wave latent period was shorter (P<0.01), the number of contraction waves in 90 s increased (P<0.01), the immunoreactive expression of D 2 receptor in colon decreased (P<0.01). Compared with the model group, the Bristol fecal scores of the Tianshu and Neiguan groups decreased (P<0.01), the 1st contraction wave latent periods were longer (P<0.01), the numbers of contraction waves in 90 s decreased (P<0.01), the positive expressions of D 2 receptor in colon increased (P<0.01, P<0.05). Compared with the Tianshu group, the immunoreactive expression of D 2 receptor in the Neiguan group decreased (P<0.01). CONCLUSION: EA at ST 25 and PC 6 can improve the symptoms of colonic motility in IBS rats. The effect of EA at ST 25 is better, which indicates that different meridians and different acupoints play specific effects.

[Effect of Electroacupuncture at "Neiguan" (PC 6) on Skin Microcirculation Blood Perfusion Volume of Heart-related Acupoints and Heart-irrelevant Acupoints in Myocardial Ischemia Rats].

OBJECTIVE: To observe the effect of low-frequency or high-frequency electroacupuncture (EA) stimulation of "Neiguan" (PC 6) on changes of skin blood perfusion volume of heart-related and heart-irrelevant acupoints in rats with normal or ischemic myocardium. METHODS: Wistar rats were randomly divided into the control, sham-operation, model, low-frequency EA and high-frequency EA groups (n=8 in each group). The myocardial ischemic (MI) model was established by occlusion of the anterior descending branch of the left coronary artery. 2 Hz and 100 Hz EA was respectively applied to left PC 6 for 20 min, once daily, a total of three times. Electrocardiogram (ECG) was recorded after the last treatment. Serum cardiac troponin T (cTnT) contents were assayed by ELISA. The skin blood perfusion volumes of bilateral PC 6, "Zusanli" (ST 36) and "Yanglingquan" (GB 34) were observed by laser speckle contrast imaging. RESULTS: In comparison with the control and sham-operation groups, the differences of "J point" and contents of serum cTnT were significantly increased in the model group (P<0.01). After EA, the differences of "J point" and contents of serum cTnT were significantly decreased in both low-frequency and high-frequency EA groups (P<0.01, P<0.05). Compared with the control group, the skin blood perfusion volumes of bilateral PC 6 and ST 36 decreased obviously in rats of the model group (P<0.01). After EA, the skin blood perfusion volumes were markedly up-regulated in the areas of PC 6 and ST 36 (P<0.01). There were no significant changes in that of GB 34 among the 5 groups (P<0.05). CONCLUSIONS: Skin blood perfusion volumes around PC 6 and ST 36 can specifically reflect the change of the myocardial state.

[Effect of Electroacupuncture at "Yintang" (GV 29) and "Tianshu" (ST 25) on the Ethology and the Expression of TRPV 1 Receptor in Colon of Irritable Bowel Syndrome Rats].

OBJECTIVE: To observe the ethology and expression of transient receptor potential vanilloid-1(TRPV 1) in colon of irritable bowel syndrome (IBS) rats treated by electroacupuncture (EA) at "Yintang" (GV 29) and "Tianshu" (ST 25), so as to explore the different effectiveness of different acupoints and its related mechanism underlying improvement of the symptoms of somatopsychic illness. METHODS: Thirty-two neonatal Wistar rats were randomly divided into blank control group, model group, Yintang (GV 29) group, and Tianshu (ST 25) group (n=8 in each group). IBS model was established by neonatal maternal separation and acetic acid enema combined with colon and rectum expansion stimulation. Rats of the GV 29 and ST 25 groups were given EA treatment at age of 9 weeks old, 20 minutes, once every other day for 5 times. Latency of the 1st time contraction wave and numbers of contractive wave in 90 s were recorded to evaluate abdominal visceral sensitivity by abdominal withdrawal reflex. Horizontal and vertical movements were observed to assess the emotional and psychological behavior of rats by open field test. TRPV 1 expression in colon was detected by immunohistochemistry method. RESULTS: Compared with the black control group, latency of the 1st time contractive wave was significantly shortened and the wave numbers in 90 s were increased significantly in the model group (P<0.01), while the numbers of vertical and horizontal movements were also significantly reduced (P<0.01), accompanied with increased TRPV 1 expression in the colon (P<0.05). Compared with the model group, After EA intervention, the levels of the above-mentioned 5 indexes were significantly reversed in the GV 29 and ST 25 groups (P<0.01,P<0.05), except the number of horizontal movement in the ST 25 group (P>0.05). The expression level of TRPV 1 was significantly lower in the ST 25 group than in the GV 29 group (P<0.01). CONCLUSIONS: EA at GV 29 and ST 25 can alleviate the abdominal pain and improve the mental and emotional disorders in IBS rats. GV 29 has a better effect on relieving the depressive-like psychoemotional behavior of IBS rats, while ST 25 is more effective in treating abdominal pain. There is significantly different influence on TRPV 1 expression in colon between these two treatment groups, which may contribute to their different effect in pain relieving.

Effects of electroacupuncture intervention on expression of cyclooxygenase 2 and microglia in spinal cord in rat model of neuropathic pain.

OBJECTIVE: To investigate the effect of electroacupuncture (EA) treatment on the expression of cyclooxygenase (COX) 2 and microglia in spinal cord by using rat model of neuropathic pain, and to probe into the relationship between COX 2 and microglia. METHODS: The rats were randomly divided into 6 groups, including normal control group, model group, sham group, EA 1 group (distant acupoints + local acupoints), EA 2 group (local acupoints), and EA 3 group (distant acupoints). Thermal withdrawal latencies were evaluated at 1 day preoperatively and 3, 5 and 7 days postoperatively. At 7 days postoperatively, the spinal COX 2 mRNA was detected by reverse-transcription polymerase chain reaction. Double immunofluorescent staining technology was applied to screen and verify the relationship between altered COX 2 and microglia. RESULTS: Compared with the model group, thermal withdrawal latencies increased after EA treatment (P<0.01). The expressions of COX 2 mRNA were up-regulated in spinal cord of rat on day 7 after surgery (P<0.05). Compared with the model group, EA stimulation (EA 1 and EA 2 groups) reversed the up-regulation of COX 2 mRNA expression (P<0.05). EA 1 and EA 2 groups might have better treatment effect compared with the EA 3 group. Fluorescent images displayed COX 2 and microglia expressed at common areas. CONCLUSIONS: EA was effective in analgesic and anti-inflammatory. EA has decreased the expression of spinal COX 2 mRNA in the trend of the therapeutic effect of "distant acupoints + local acupoints", and "local acupoints" intervention may be superior to that of "distant acupoints" intervention. Microglia may be related to the formation of COX 2.

Effects of acupuncture applied to sanyinjiao with different stimuli on uterine contraction and microcirculation in rats with dysmenorrhea of cold coagulation syndrome.

In this study, we try to evaluate the effects of acupuncture stimulation with different amounts at Sanyinjiao (SP6) on uterine contraction and uterus microcirculation in rats with dysmenorrhea of cold coagulation syndrome and to explore whether there is direct relativity between "De qi" and needle stimulus intensity. Diestrus female rats were randomly divided into 4 groups, including saline control group, model control group, "A" stimulus group (with strong stimulus), and "B" stimulus group (with weak stimulus). We found that dysmenorrhea rats of the cold coagulation syndrome present a high intensity in uterine tension and high contraction of microvascular diameter. Acupuncture applied with two different stimuli could relieve the symptoms, but, compared with "B" stimulus, "A" stimulus leads to better outcomes on reducing uterine contraction and increasing diameter of uterine microvascular; moreover, hand manipulation during needling mediates the curative effect on the microvascular diameter. Our finding indicates that using thick needles and deep insertion with hand manipulation are more effective and achieve desired level of "De qi" in dysmenorrhea rats.

Effects of preventive acupuncture and moxibustion on fat accumulation, blood lipid, and uterus e 2 of menopause rats.

Objective. To observe the effect of preventive acupuncture and moxibustion on blood lipid of menopause rats. Methods. Seventy 10-month-old SD rats with estrous cycle disorders were divided into three control groups and four treatment groups (n = 10/group) and another ten 3.5-month-old female SD rats were chosen as young control group. Preventive acupuncture and moxibustion were applied at Guanyuan (CV 4). Body weight growth rate has been recorded. Plasma total cholesterol (TC), triglyceride (TG), low density lipoprotein (LDL), and high density lipoprotein (HDL) levels and uterus E 2 level were measured. Results. Compared to young control group, plasma TC and LDL increased and uterus E 2 reduced significantly in 12-month-old control group. Compared to 12-month-old control group, plasma TC and LDL level and body weight growth rate decreased while HDL level increased remarkably in preventive acupuncture 12-month-old group. Compared to 14-month-old control group, plasma TC level and body weight growth rate decreased remarkably in preventive moxibustion 14-month-old group. Conclusions. Preventive acupuncture and moxibustion can significantly decrease the plasma TG and LDL, increase the plasma HDL, and prevent fat accumulation. Our finding suggests that preventive acupuncture and moxibustion have beneficial effects on blood lipid. Different treatment effects were found between preventive acupuncture and preventive moxibustion.

Acupuncture De-qi: From Characterization to Underlying Mechanism.

De-qi refers to the participant's subjective sensations and objective body responses as well as the acupuncturist's perceptions while the acupuncturist needles certain acupoints in the participant's body. In recent years, De-qi is getting increasing attention of the researchers and many efforts have been made to understand its mechanism. By the broad literature survey, this paper explores the subjective De-qi sensation of the patients, its influencing factors, and the resulting physiological responses. The purpose of this paper is expected to find out a possible mechanism of De-qi and to provide certain scientific evidence for acupuncture fundamental research and clinical practice.

[Effects of electroacupuncture on pain reactions, expression of spinal kappa-opioid receptor and contents of enkephalin and beta-endorphin in periaqueductal gray of midbrain in dysmenorrhea model rats].

OBJECTIVE: To observe effects of electroacupuncture (EA) on the expression of kappa-opioid receptor in the dorsal horn of the spinal cord and contents of enkephalin(ENK) and beta-endorphin (beta-EP) in the periaqueductal gray (PAG) of midbrain in dysmenorrheal rats, so as to reveal its underlying mechanism in relieving dysmenorrhea. METHODS: A total of 80 female SD rats were randomized into saline control (control), model, Sanyinjiao (SP 6), Xuanzhong (GB 39), non-acupoint groups (16 rats/group). Dysmenorrhea model was established by subcutaneous injection of estradiol benzoate (0.5 mg/rat on the 1st day and 10th day, 0.2 mg/rat from the 2nd day to the 9th day). One hour after the last injection, oxytocin (2 U/rat) was given intraperitoneally, for rats of the control group, the same dose of saline was given (i. p.). On the 10th day, EA (2 Hz/100 Hz, 0.1-0.3 mA) was applied to "Sanyinjiao" (SP 6), "Xuanzhong" (GB 39) and non-acupoint (the mid-point between the Stomach Meridian and Gallbladder Meridian, and in parallel with GB 39) for 20 min, respectively. Latency and number of writhing response, and writhing score (according to Schmauss's and Yaksh's method) were recorded. The expression of kappa-opioid receptor (kappa-OR) in T13, L1 , L2, L6 and S1 segments of spinal cord was determined by immunohistochemistry, and the contents of ENK and beta-EP in the midbrain PAG were assayed by ELISA. RESULTS: (1) Compared with the saline control group, the writhing latency of the model group was significantly shortened (P < 0.01), while the writhing times and writhing score of the model group were increased significantly (P < 0.01). Compared with the model group, the writhing latency of SP 6 group was significantly prolonged (P < 0.05), while the writhing scores and writhing times of the SP 6, GB 39 and the non-acupoint groups decreased significantly (P < 0.01). (2) In comparison with the control group, kappa-OR expression in the dorsal horn of L2 segment of spinal cord was upregulated significantly in the model group (P < 0.05). Compared to the model group, kappa-OR expression levels in the dorsal horns (DHs) of spinal T13, L1, L2, L6 and S1 segments in the SP 6 group were upregulated significantly (P < 0.01). ENK and beta-EP contents of PAG in the SP 6 and GB 39 groups were increased considerably (P < 0.01, P < 0.05). The effects of SP 6 were significantly superior to those of GB 39 in upregulating kappa-OR expression of spinal L1, L2 and L6 DHs and in upregulating beta-EP content of PAG; and superior to non-acupoint in upregulating kappa-OR expression of spinal T13, L1, L2, L6 and S1 DHs and in increasing both ENK and beta-EP contents of PAG (P < 0.01, PF < 0.05). No significant differences were found between the non-acupoint group and the model group in writhing latency, kappa-OR expression levels of spinal T13, L1, L2 and S1 DHs, and in ENK and beta-EP contents of PAG (P > 0.05). CONCLUSION: EA of SP 6 can significantly alleviate pain reactions in dysmenorrhea rats, which is closely associated with its functions in upregulating spinal kappa-OR expression and ENK and beta-EP contents in PAG. EA of SP 6, GB 39 and non-acupoint has some different degrees of efficacies in relieving dysmenorrhea and in upregulating spinal K-OR expression.

[Effect of electroacupuncture at different acupoints on plasma TXB2 and 6-keto-PGF(1alpha), contents in dysmenorrhea rats].

OBJECTIVE: To observe the effect of electroacupuncture (EA) on plasma thromboxane B2(TXB2) and 6-keto-prostaglandin F1alpha (6-keto-PGF1alpha) levels in dysmenorrhea rats in order to investigate its mechanism underlying relief of primary dysmenorrhea and specificity of acupoint efficacy. METHODS: Female SD rats with diestrus were randomly divided into saline control (control), model, EA Sanyinjiao (SP 6), EA Xuehai (SP 10), EA Xuanzhong (GB 39) and EA non-acupoint (NAP) groups, with 10 rats in each. Dysmenorrhea model was established by subcutaneous injection of Estradiol Benzoate (0.5 mg/rat on the 1st and 10th day, and 0.2 mg/rat from the 2nd to the 9th day) and intraperitoneal injection of Oxytocin (0.2 mL/rat, 1 h after last injection of Estradiol Benzoate on the 10th day). EA was applied to bilateral SP 6, SP 10, GB 39, and non-acupoint (the mid-point between the Gallbladder and Stomach meridian at the GB 39 level) for 20 min. The latency and score of writhing were recorded for 20 min. Plasma TXB2 and 6-keto-PGF1alpha contents were detected by radioimmunoassay. RESULTS: Compared with the control group, the latency of writhing in the model group was shortened considerably (P < 0.01), and the writhing score was increased significantly (P < 0.01). In comparison with the model group, the writhing latency was increased significantly only in the EA-SP 6 group (P < 0.05), and the writhing scores in the EA-SP 6, EA-SP 10, EA-GB 39 and EA-NAP groups were reduced remarkably (P < 0.01). Plasma TXB2 content and the ratio of TXB2/6-keto-PGF1alpha. were significantly higher in the model group than in the control group (P < 0.01). Compared to the model group, plasma TXB2 levels and the ratios of TXB2/6-keto-PGF1alpha. in the EA-SP 6, EA-SP 10, EA-GB 39 and EA-NAP groups were downregulated markedly (P < 0.05, P < 0.01), while plasma 6-keto-PGF1alpha was upregulated strikingly only in the EA-SP 6 group (P < 0.05). No significant differences were found among the EA-SP 6, EA-SP 10, EA-GB 39 and EA-NAP groups in the writhing latency and writhing score, plasma TXB2 and 6-keto-PGF1alpha, levels (P > 0.05). CONCLUSION: EA can relieve pain reaction in dysmenorrhea rats, which may be closely associated with its effects in downregulating plasma TXB2, upregulating plasma 6-keto-PGF1alpha, content, and balancing plasma TXB2/6-keto-PGF1alpha. The effect of EA of SP 6 is relatively better.

[Affection of acupotomy lysis on leu-enkephalin (L-ENK) content in different parts of centrum of rats with knee osteoarthritis].

OBJECTIVE: To study the analgesia mechanism of needle-knife lysis in spinal cord and other parts of central nervous system by comparing the changes of Leu-Enkephalin (L-ENK) content in different parts of centrium of rats undergone needle-knife lysis and electro-acupuncture respectively. METHODS: Sixty healthy SD rats were randomly devided into normal control group, model group, needle-knife lysis (NKL) group and electro-acupuncture (EA) group. 4% papain solution mixed with 0.3 mol/L cysteine solutin in the ratio of 1:1, paused for 0.5 h,injected the mixture, 20 microl each time,into the left knee joint cavities of rats in model, NKL, EA groups at the 1st, 4th, 7th day. After 4 weeks in NKL group and EA group were treated with needle-knife lysis and electro-acupuncture, respectively. Three weeks after treatment, samples of spinal cord of the swollen part of rat waists and rat brains were taken from and the content of L-ENK of medulla oblongata, midbrain, pituitary gland, and hippocampus were measured. RESULTS: L-ENK content of model group increased higher than that of normal control group in spinal cord, hippocampus and midbrain (P < 0.01); there were no significant difference between normal control group and modle group on L-ENK in medulla oblongata and thalamus (P > 0.05). After intervening of NKL or EA, L-ENK content of NKL group increased higher in hippocampus than that of model group and EC group (P < 0.05); but L-ENK content of NKL group in midbrain was lower than that of model group (P < 0.05). CONCLUSION: Needle-knife lysis has characteristic of regulation for the L-ENK content in different parts of central nervous system of rats with knee osteoarthritis, and analgesic effect of needle-knife was possibly related with regulation of center L-ENK.