To observe the angiogenesis effect of electroacupuncture (EA) at Shuigou acupoint (GV 26) in the treatment of cerebral ischemia, and explore the value of miRNA-7 (miR-7) in it. First, 48 mice were randomly divided into sham operation, middle cerebral artery occlusion (MCAO) model, and EA treatment groups. Then 9 mice were divided into carrier control group, miR-7 knockout group and miR-7 overexpression group (n=3 each group). Finally, 20 mice were divided into model and carrier control group, model and miR-7 knockout group, EA treatment and carrier control group and EA treatment and miR-7 overexpression group, with 3–6 mice in each group. The MCAO model was established in the MCAO and EA groups. Neurological deficit score and 2,3,5-triphenyltetrazolium chloride (TTC) staining were used to evaluate the severity of cerebral ischemia. Hematoxylin-eosin staining was used to describe basic pathological changes. Immunohistochemistry was used to quantify cerebral microvessel density. Real-time PCR and Western blot were used to detect the expression of miR-7 and its downstream target genes Krüppel-like factor 4/vascular endothelial growth factor (KLF4/VEGF) and angiopoietin-2 (ANG-2) in the ischemic cerebral cortex. After EA, neurological deficit scores and infarction volumes decreased, and the density of cerebral microvessels increased. In the MCAO group, miR-7 expression was higher than that in the sham group (P<0.01). After EA at GV 26, miR-7 expression decreased (P<0.01) and the expression of downstream target genes KLF4/VEGF and ANG-2 increased as compared with the MCAO group (P<0.01). After EA combined with overexpression of miR-7, the expression of downstream target genes KLF4/VEGF and ANG-2 decreased compared to the control EA group (P<0.01). After miR-7 knockdown, the expression of KLF4/VEGF and ANG-2 increased (P<0.05 or P<0.01). EA could promote angiogenesis in MCAO mice likely by inhibiting the expression of miR-7 and relieving inhibition of downstream target genes KLF4/VEGF and ANG-2.

OBJECTIVES:To observe the effect of electroacupuncture on miR-142-5p and ADAMTS1/PI3K/AKT pathway in rats with ischemic stroke, so as to explore the regulatory mechanism of electroacupuncture on angiogenesis after ischemic stroke. METHODS:This study was divided into two parts. The first part of the experiment：SD rats were randomly divided into sham operation group, model group and electroacupuncture group. There were 20 rats in each group. The middle cerebral artery occlusion (MCAO) rat model was prepared using a modified Longa's method. In the electroacupuncture group, "Shuigou" (GV26) was selected for electroacupuncture intervention (4 Hz/20 Hz) for 30 min each time. The rats in the electroacupuncture group were given electroacupuncture immediately after successful modeling, once a day for 4 times. Hunter score and TTC staining were used to observe the neurological deficits and infarct volumes respectively；HE staining was used to observe the cortical pathological changes；immunohistochemistry was used to determine the changes of cerebral microvascular density. Real-time quantitative PCR and Western blot were used to observe the miR-142-5p expression, mRNA and protein expression levels of ADAMTS1, VEGF, PI3K, AKT, eNOS in ischemic cortex. The second part of the experiment：The rats were randomly divided into electroacupuncture+control group and electroacupuncture+miR-142-5p Antagomir group with 8 rats in each group. MCAO model was established after injection. Electroacupuncture+control group was given 0.9% sodium chloride solution injected into the right ventricle.The rats in the electroacupuncture+miR-142-5p Antagomir group were injected with miR-142-5p inhibitor into the right ventricle 30 min before modeling. Rats in electroacupuncture+control group and electroacupuncture+miR-142-5p Antagomir group were all given the same electroacupuncture treatment. Real-time fluorescence quantitative PCR was used to observe the effect of miR-142-5p Antagomir on the expression of miR-142-5p and ADAMTS1 mRNA. The effect of miR-142-5p Antagomir on ADAMTS1 protein was observed by Western blot. RESULTS:In the first part of the experiment, compared with the sham operation group, the Hunter score in the model group was significantly increased (P<0.01)；the volume of cerebral infarction in the model group was significantly increased (P<0.01)；the degree of brain edema and neuronal necrosis and the density of cerebral microvessels was increased；the cerebral microvascular density was significantly increased (P<0.01)；the expression levels of miR-142-5p and the mRNA expression levels of VEGF, AKT and eNOS were significantly decreased (P<0.01, P<0.05), and the protein expression levels of VEGF, p-AKT and eNOS were significantly down-regulated (P<0.01), while the mRNA expression levels of ADAMTS1 and PI3K, and the protein expression levels of ADAMTS1 and p-PI3K were all up-regulated (P<0.01, P<0.05) in the model group. Compared with the model group, after intervention, the Hunter score in the electroacupuncture group was decreased (P<0.01), the volume of cerebral infarction was significantly decreased (P<0.01)；the degree of brain edema and neuronal necrosis were alleviated；the cerebral microvascular density was significantly increased (P<0.01)；the expression of miR-142-5p and the mRNA expression of VEGF, PI3K, AKT and eNOS were increased (P<0.01), the protein expressions of VEGF, p-PI3K, p-AKT and eNOS were increased (P<0.01, P<0.05), while the mRNA and protein expression of ADAMTS1 were decreased (P<0.05, P<0.01). After injection of miR-142-5p inhibitor, compared with electroacupuncture+control group, the expression of miR-142-5p in electroacupuncture+miR-142-5p Antagomir group was decreased(P<0.05), while the mRNA and protein expression of ADAMTS1 were increased (P<0.01, P<0.05). CONCLUSIONS:Electroacupuncture at GV26 can improve the neurological damage of ischemic stroke rats, reduce the volume of cerebral infarction and promote angiogenesis. The mechanism may be associated with the function of electroacupuncture in promoting the expression of miR-142-5p, so as to inhibit the expression of its target gene ADAMTS1, mediate the up-regulation of VEGF expression, activate PI3K/AKT pathway, promote the release of eNOS, and participate in promoting angiogenesis in ischemic stroke rats.

缺血性脑卒中指脑血循环障碍导致脑血管堵塞或严重狭窄,使脑血流灌注下降,进而缺血、缺氧导致脑血管供血区脑组织死亡,已成为全球第二大致死原因.缺血性脑卒中及其后遗症对病人身心健康造成危害,故对缺血性脑卒中的治疗问题亟待解决.非编码RNA在中医药诊治机制中发挥着重要作用.通过文献整理的方法综述中医药调控非编码RNA干预缺血性脑卒中机制的研究进展,旨在为临床对缺血性脑卒中的诊断、辨证论治、治疗方案拟定、新药研发等提供参考.

Neuropathic pain (NP) is directly caused by lesions or diseases of the somatosensory system. It can be attributed to a variety of causes and has a complex pathological mechanism. Purinergic P2X receptors play a critical role in NP. P2X receptors will be activated by the extracellular adenosine triphosphate (ATP) to mediate the pain when the tissue is damaged. In vertebrates, there are 7 subtypes of P2X receptors (P2X1-7) encoded by genes, among which the receptors of P2X3, P2X4, and P2X7 are closely related to NP. Acupuncture produces good therapeutic efficacy on NP. To explore the mechanism of purinergic signaling involved in acupuncture analgesia, a review of the actions of P2X3, P2X4, and P2X7 receptors in acupuncture for NP was conducted. The P2X3 receptor is specifically expressed in small- and moderate-sized neurons in the dorsal root ganglion of the primary afferent nerve and trigeminal ganglion. Acupuncture can directly mediate analgesia by down-regulating the expression of P2X3 receptor in neurons. P2X4 and P2X7 receptors mainly exist in the spinal microglia. Acupuncture can inhibit the nociceptive transmission of substantia gelatinosa neurons by regulating P2X4 receptor in spinal microglia. Moreover, acupuncture may alleviate NP by inhibiting the secretion of inflammatory factors by reducing P2X7 receptor in the spinal microglia.

环状RNA(circRNA)是在近些年被高度关注的一种非编码RNA.越来越多的研究结果显示,circRNA在缺血性脑卒中发病机制中发挥了重要作用,并有望成为缺血性脑卒中治疗靶点和临床检测标志物.文章就circRNA与缺血性脑卒中相关研究现状进行综述.

目的:探讨后疫情时代同伴教学法(PI)在刺法灸法学混合式教学中的应用效果.方法:以针灸推拿专业52名学生为研究对象,随机分为对照组和观察组.对照组采用传统教学+线上教学模式,观察组在此基础上增加同伴教学.课程结束后,比较两班学生的理论知识和实践技能考核成绩,调查同伴教学效果.结果:观察组总成绩为(89.08±2.97)分,高于对照组的(81.95±4.12)分(P＜0.01).观察组学生的理论考核和操作考核成绩也均显著高于对照组(P＜0.01).同伴教学有助于激发学生的学习兴趣,养成独立思考能力、思辨能力、培养合作意识,增强师生、学生间的互动交流,互相学习.结论:同伴教学法在刺法灸法学课程教学中的应用效果较好.