Objective To evaluate the role of microRNA9 (miR-9) in spinal dorsal horn neurons in over-expression of calcium homeostasis modulator 1 (CALHM1) in rats with diabetic neuropathic pain (DNP).Methods Ninety-three healthy male Sprague-Dawley rats,aged 2 months,weighing 180-200 g,were used in the study.Diabetes mellitus was induced by intraperitoneal 1％ streptozocin (STZ) 60 mg/kg.The experiment was performed in two parts.Experiment Ⅰ The rats were divided into control group (group C,n=10) and DNP group (n=83) using a random number table.The mechanical paw withdrawal threshold (MWT) was measured before STZ injection and at 1,2,3,4,5 and 6 weeks after STZ injection.The expression of miR-9 in the spinal dorsal horn was determined using the in situ hybridization at 6 weeks after STZ injection.Experiment Ⅱ The rats with DNP were selected at 6 weeks after STZ injection,and the spinal dorsal horn neurons were isolated and cultured.The neurons were seeded in culture plates at the density of 5×106 cells/ml (2 ml/well) and divided into 2 groups (n=18 each) using a random number table:control group (group C) and miR-9 antisense oligonucleotide group (group ASO).The neurons were cultured in normal culture atmosphere in group C.In group ASO,the single nucleotide sequence of miR-9 antisense oligonucleotide sequence 5'-UUCUCCGAACGUGUCACGUTT-3'was added with the final concentration of 100 pmol/L.The expression of miR-9 and CALHM1 mRNA was detected using quantitative real-time polymerase chain reaction at 48 h of incubation.The expression of CALHM1 was detected by Western blot at 72 h of incubation.Results Experiment Ⅰ Compared with group C,the MWT was significantly decreased at 2-6 weeks after STZ injection,and the expression of miR-9 in the spinal dorsal horn was up-regulated in group DNP (P＜0.05).Experiment Ⅱ Compared with group C,the expression of miR-9 and CALHM1 protein and mRNA in spinal dorsal horn neurons was significantly down-regulated in group ASO (P＜0.05).Conclusion miR-9 in spinal dorsal horn neurons probably mediates the over-expression of CALHM1 in rats with DNP.

BACKGROUNDPrevious studies demonstrated that vagus nerve stimulation (VNS) is an effective therapy for drug-resistant epilepsy. Acupuncture is also used to treat epilepsy. This study was designed to examine the safety and effectiveness of transcutaneous auricular vagus nerve stimulation (ta-VNS) for patients with drug-resistant epilepsy.

METHODSA total of 50 volunteer patients with drug-resistant epilepsy were selected for a random clinical trial to observe the therapeutic effect of ta-VNS. The seizure frequency, quality of life, and severity were assessed in weeks 8, 16, and 24 of the treatment according to the percentage of seizure frequency reduction.

RESULTSIn the pilot study, 47 of the 50 epilepsy patients completed the 24-week treatment; three dropped off. After 8-week treatment, six of the 47 patients (12%) were seizure free and 12 (24%) had a reduction in seizure frequency. In week 16 of the continuous treatment, six of the 47 patients (12%) were seizure free; 17 (34%) had a reduction in seizure frequency. After 24 weeks' treatment, eight patients (16%) were seizure free; 19 (38%) had reduced seizure frequency.

CONCLUSIONSimilar to the therapeutic effect of VNS, ta-VNS can suppress epileptic seizures and is a safe, effective, economical, and widely applicable treatment option for drug-resistant epilepsy. (ChiCTR-TRC-10001023).

Adolescent ; Adult ; Epilepsy ; therapy ; Female ; Humans ; Male ; Transcutaneous Electric Nerve Stimulation ; methods ; Treatment Outcome ; Vagus Nerve Stimulation ; methods ; Young Adult