[Effect of Yes-associated protein in regulation of electroacupuncture pretreatment on cerebral ischemia reperfusion injury rats].

OBJECTIVE: To observe the effect of electroacupuncture (EA) pretreatment on inflammatory reaction, apoptosis and expression of Yes-associated protein (YAP) of ischemic penumbra of cerebral cortex in cerebral ischemia reperfusion injury rats, and to explore the possible mechanism of its neuroprotection effect. METHODS: A total of 84 SD rats were randomized into a sham operation group (12 rats), a model group (18 rats), an EA group (18 rats), an EA+YAP virus transfection group (18 rats) and an EA+virus control group (18 rats). Except for the sham operation group, thread embolization method was adopted to establish the middle cerebral artery occlusion (MCAO) model in rats of the other groups. EA was applied at "Baihui" (GV 20) and "Dazhui" (GV 14) for 30 min in the 3 EA intervention groups 2 h before model establishment, disperse-dense wave, 2 Hz/15 Hz in frequency and 1 mA in intensity. Adenovirus transfection technique was used to induce gene silencing of YAP in the EA+YAP virus transfection group, and adenovirus vectors was injected as negative control in the EA+virus control group 4 d before model establishment. Twenty-four hours after model establishment, neurological function score was evaluated, the relative cerebral infarction area was observed by TTC staining, the apoptosis in the ischemic penumbra of cerebral cortex was detected by TUNEL staining, the levels of inflammatory factors IL-1ß, IL-6 and TNF-α in the ischemic penumbra of cerebral cortex was detected by ELISA method, the expression of YAP was detected by Western blot and immunofluorescence. RESULTS: Compared with the sham operation group, the expression of YAP was increased in the model group (P<0.05); compared with the model group, the expression of YAP in the ischemic penumbra of cerebral cortex was increased in the EA group (P<0.05). Compared with the sham operation group, the neurological function score, the percentage of TUNEL positive cells and the levels of IL-1ß, IL-6 and TNF-α in the ischemic penumbra of cerebral cortex were increased in the model group (P<0.001, P<0.01); compared with the model group, the neurological function score, the relative cerebral infarction area, the percentage of TUNEL positive cells and the levels of IL-1ß, IL-6 and TNF-α in the ischemic penumbra of cerebral cortex were decreased in the EA group (P<0.05, P<0.01); compared with the EA group, the neurological function score, the relative cerebral infarction area, the percentage of TUNEL positive cells and the levels of IL-1ß, IL-6 and TNF-α in the ischemic penumbra of cerebral cortex were increased in the EA+YAP virus transfection group (P<0.01, P<0.05); compared with the EA+YAP virus transfection group, the neurological function score, the relative cerebral infarction area, the percentage of TUNEL positive cells and the levels of IL-1ß, IL-6 and TNF-α in the ischemic penumbra of cerebral cortex were decreased in the EA+virus control group (P<0.01, P<0.05). CONCLUSION: Electroacupuncture pretreatment can effectively improve the ischemia reperfusion injury, its mechanism may be related to up-regulating the expression of YAP in the ischemic penumbra of cerebral cortex and relieving the apoptosis and inflammatory reaction.

Role of spinal adenosine A1 receptors in the analgesic effect of electroacupuncture in a rat model of neuropathic pain.

OBJECTIVE: The aim of this study was to determine the role of spinal adenosine A1 receptors (A1Rs) in the analgesic effects of electroacupuncture (EA) for neuropathic pain. METHODS: We performed EA for 30 minutes at the zusanli acupoint in the legs of rats with previously induced chronic constriction injuries and observed the mechanical and thermal pain thresholds 1 hour later. We also examined adenosine levels by high-performance liquid chromatography and A1R expression in the L4-6 spinal cord by western blot analysis. We then injected A1R short interfering RNA (AV-shA1RNA) into the L4-6 spinal cord to downregulate A1R expression and re-examined the mechanical and thermal pain thresholds. RESULTS: Adenosine levels and A1R expression in the L4-6 spinal cord were increased at 1 hour after EA. In addition, EA exhibited an analgesic effect that was reversed by AV-shA1RNA. CONCLUSIONS: Our results suggest that EA at the zusanli acupoint elicits an analgesic effect against neuropathic pain, mediated by A1Rs in the spinal cord.

Identifying the Genes Regulated by AtWRKY6 Using Comparative Transcript and Proteomic Analysis under Phosphorus Deficiency.

Phosphorus (P) is an important mineral nutrient for plant growth and development. Overexpressing AtWRKY6 (35S:WRKY6-9) was more sensitive and wrky6 (wrky6-1) was more resistant under low Pi conditions. To better understand the function of AtWRKY6 under low phosphate stress conditions, we applied two-dimensional gel electrophoresis (2-DE) to analyse differentially expressed proteins in the shoots and roots between wild type, 35S:WRKY6-9 and wrky6-1 after phosphorus deficiency treatment for three days. The results showed 88 differentially abundant protein spots, which were identified between the shoots and roots of 35S:WRKY6-9 and wrky6-1 plants. In addition, 59 differentially expressed proteins were identified in the leaves and roots of 35S:WRKY6-9 plants. After analysis, 9 genes with W-box elements in their promoter sequences were identified in the leaves, while 6 genes with W-box elements in their promoter sequences were identified in the roots. A total of 8 genes were identified as potential target genes according to the quantitative PCR (QPCR) and two dimension difference gel electrophoresis, (2D-DIGE) results, including ATP synthase, gln synthetase, nitrilase, 14-3-3 protein, carbonic anhydrases 2, and tryptophan synthase. These results provide important information concerning the AtWRKY6 regulation network and reveal potential vital target genes of AtWRKY6 under low phosphorus stress. two dimension difference gel electrophoresis, 2D-DIGE.

[Effect of transcutaneous acupoint electrical stimulation on stress in brain surgery with propofol target controlled infusion general anesthesia].

OBJECTIVE: To study the effect of transcutaneous acupoint electrical stimulation (TAES) on stress who received propofol target controlled infusion (TCI) general anesthesia in brain surgery. METHODS: Totally 40 neurosurgical patients of I-II grade (ASA grading) in our hospital were randomly divided into the TAES group (T group) and the control group (C group), 20 in each group. All patients received intravenous anesthesia by propofol TCI. The TAES intervention was adopted in those of C group. Electrodes were only applied to corresponding acupoints without electric stimulation. The arterial blood was withdrawn before TAES (T0), before anesthesia (T1), before cutting (T2), at 60 min after encephalic incision (T3), immediately after incisions suture (T4), at about 10 min after removing tracheal catheters (T5) to detect beta-endorphin (beta-EP), cortisol (COR), adrenalin (E), blood sugar (Glu). The heart rate (HR) and mean arterial pressure (MAP) were recorded. The total time of surgery, anesthesia, total infusion amount, blood lost amount, and urine amount were recorded. RESULTS: In both groups, HR, MAP, COR, and E at T2 were lower than at T0 significantly (P < 0.05). beta-EP in group C at T2 was lower than at T0 significantly (P < 0.05). HR, MAP, COR in group C at T3 were higher than at T0 significantly (P < 0.05). HR, MAP, E, and Glu in group C at T4 and T5 were higher than at T0 significantly (P < 0.05). beta-EP in group T at T1 and T3 were higher than at T0 significantly (P < 0.05). HR, COR, E, Glu, and beta-EP in group T at T4 and T5 were higher than at T0 significantly (P < 0.05). Between groups, comparing with the time point T0, the amplitude of variation of MAP, COR, and E at T2 in group C were significantly less (P < 0.05); the amplitude of variation of HR, MAP, and COR at T3 in group C were less significantly, when compared with the time point T0 (P < 0.05); the amplitude of variation of HR, MAP, COR, E, and Glu at T4 and T5 in group C were less significantly, when compared with the time point T0 (P < 0.05). When comparing the two groups, the amplitude of variation of beta-EP at time points of T1, T3, T4, and T5 in group T were larger than at T0 in group C (P < 0.05). CONCLUSION: TAES could reduce stress and stabilize the internal environment when used in brain surgery with propofol TCI general anesthesia.