Objective To investigete the effect of low-concentration ozone (O3) on voltage-gated calcium channels in fetal rat spinal cord neurons in vitro. Methods The spinal cord neurons were cultured in vitro and identified by immunohistochemistry staining. Then the cells were seeded into the culture dish and randomly divided into 3 groups: control group (group C, n = 5), O3 15 μg/ml (group O3-15, n = 4) and O3 20 μg/ml group (group O3-20, n = 8). The cells were exposed to O3 15 and 20 μg/ml in group O3-15 and O3-20 respectively, while the cells were exposed to air in group C. After 20 min incubation, the electrophysiological activity of calcium channels in neurons was recorded using whole-cell patch-clamp technique. Results Peak calcium current density was significantly increased in group O3-15 and O3-20 compared with group C ( P ＜ 0.05). Half-activation voltage was significantly lower in group O3-15 and O3-20 than in group C ( P ＜ 0.05). Peak calcium current density was significantly higher in group O3 -20 than in group O3-15 ( P ＜ 0.05) . There was no significant difference in half-inactivation voltage among the three groups ( P ＞ 0.05). Conclusion Low concentration of O3 can make the activation of the calcium ion channel easy in fetal rat spinal cord neurons in vitro.

Objective To investigate the effects of ozone (O3) on Wnt/β-catenin signaling pathway in the articular cartilage of rats with osteoarthritis (OA).Methods Eighteen male SPF Wistar rats,aged 3 months,weighing 200-250 g,were randomly divided into 3 groups (n =6 each) using a random number table:control group (group C),group OA,and O3 group (group O).OA was induced by injection of monosodium iodoacetate 3 mg (50 μl) into the right knee joint cavity.On 7th day after the model was established successfully,25 μg/ml O3 1 ml were injected into the knee joint cavity,once a week for 3 consecutive weeks in group O.Behavioral changes were observed after establishment of the model.At 1 day before establishment of the model,and 1,4,7,14,21 and 28 days after establishment of the model,the mechanical paw withdrawal threshold (MWT) was measured.At 28 days after establishment of the model,the total knee joint was removed and stained with haematoxylin and eosin for examination of the pathological changes of the cartilage (under light microscope) and for determination of the expression of β-catenin and matrix metalloproteinase-13 (MMP-13) in the cartilage (by immunohistochemistry).Results The signs of OA such as hind-limb motor dysfunction,knee joint swelling,or decreased joint motion,and signs of hyperalgesia such as lickings were observed after establishment of the model in rats.Compared with group C,the MWT was significantly decreased at each time point after establishment of the model,and the ex pression of β-catenin and MMP-13 in the cartilage was significantly up-regulated in the other two groups(P＜0.05).Compared with group OA,the MWT was significantly increased at 7-28 days after establishment of the model,and the expression of β-catenin and MMP-13 in the cartilage was significantly down-regulated in group O (P＜0.05).The pathological changes of the cartilage were significantly reduced in group O as compared with group OA.Conclusion The mechanism by which O3 mitigates OA is probably related to inhibition of Wnt/β-catenin signaling pathway activation in the articular cartilage in rats.

Objective To evaluate the effect of injection of ozone (O3) through lateral recess for the treatment of radiculitis caused by lumbar disc herniation on the blood glucose in patients with diabetes mellitus.Methods Ninety-six patients with radiculitis caused by lumbar disc herniation involving 48 patients with diabetes mellitus and 48 patients without diabetes mellitus were enrolled in the study.The diabetic or non-diabetic patients were randomly divided into 2 groups ( n =24 each):non-diabetic patients-O3 group (group N-O3 ),non-diabetic patients-ghcocorticoid group (group N-GC),diabetic patients-O3 group (group D-O3 ),and diabetic patients-glucocorticoid group (group D-GC).In N-O3 and D-O3 groups,30 mg/L O3 10 ml was injected via the lateral recess.In N-GC and D-GC groups,compound betamethasone injection 3.5 mg was injected via the lateral recess.The blood glucose level was measured before treatment (T1),1 and 4 h after treatment (T3,4),and 1,3 and 7 days after treatment (T5-7).VAS score was recorded at T1,immediately after treatment (T2),and at T5-7.The patients' quality of life was measured using the Medical Outcomes Study 36-Item Short-Form Health Survey (MOS SF-36 Health Survey) questionnaire at T1,7.The therapeutic effect was evaluated at T7.The side effects were recorded.Results Compared with the baseline value at T1,VAS scores were significantly decreased after treatment,while the MOS SF-36 Health Survey questionnaire scores for physical functioning,bodily pain,role emotional,and mental health in all groups,for general health in group N-GC,and for vitality in groups D-O3 and N-O3 were significantly increased at T7,and the blood glucose level was significantly increased at T4 in groups N-GC and D-O3,and at T3-7 in group D-GC ( P ＜ 0.05).There was no significant change in the blood glucose level before and after treatment in group N-O3.The blood glucose level was significantly higher in group N-GC than in group N-O3,and in group D-GC than in group D-O3 ( P ＜ 0.05).There were no significant differences in the excellent and good rates and all the MOS SF-36 Health Survey questionnaire scores among all groups.No side effects were found in the study.Conclusion Injection of ozone through lateral recess for treatment of radiculitis caused by lumbar disc herniation exerts no influence on the blood glucose level in patients with diabetes mellitus.