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Objective To evaluate the effect of dexmedetomidine pretreatment on contents of glu-tamic acid and γ-aminobutyric acid during focal cerebral ischemia-reperfusion ( I∕R) in rats. Methods Thirty-six clean-grade healthy male Wistar rats, aged 10 months, weighing 250-300 g, were divided into 3 groups using a random number table method: sham operation group (group S), I∕R group, and dexme-detomidine group (group D). Focal cerebral I∕R was induced by occlusion of the left middle cerebral artery for 30 min followed by reperfusion in anesthetized rats. The left middle cerebral artery was only isolated but not occluded in group S. Sterile normal saline 1 ml was intraperitoneally injected, and 30 min later the model of focal cerebral I∕R was established in group I∕R. Rats were sacrificed at the end of reperfusion, and brains were removed for determination of contents of glutamic acid and γ-aminobutyric acid in brain tis-sues and for examination of ultrastructure (with an electron microscope). Results Compared with group S, the content of glutamic acid was significantly increased, and the content of γ-aminobutyric acid was de-creased in I∕R and D groups (P＜0. 05). Compared with group I∕R, the content of glutamic acid was signif-icantly decreased, and the content of γ-aminobutyric acid was increased in group D (P＜0. 05). The dam-age to the ultrastructure of brain tissues was significantly attenuated in group D when compared with group I∕R. Conclusion Dexmedetomidine pretreatment can reduce focal cerebral I∕R injury through decreasing the content of glutamic acid and increasing the content of γ-aminobutyric acid in rats.
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Objective To evaluate the effect of exogenous resolvin D2 on radicular pain in a rat model of non-compressive lumbar intervertebral disc herniation.Methods Thirty-six clean-grade healthy male Sprague-Dawley rats,aged 8 weeks,weighing 230-270 g,were divided into 3 groups (n=12 each) using a random number table method:sham operation group (group S),radicular pain induced by non-compressive lumbar intervertebral disc herniation group (group P) and exogenous resolvin D2 group (group R).The right L5 dorsal root ganglions were covered by autologous nucleus pulposus tissues to establish the model of non-compressive lumbar disc herniation in P and R groups.The corresponding surgical site was only exposed in group S.The corresponding drugs were intrathecally injected within 3 days after establishing the model,phosphate buffer solution 10 μl was injected in group P,1 ng/μl resolvin D2 solution 10 μl was injected in group R,and normal saline 10 μl was given for pipe washing after administration in the three groups.The mechanical paw withdrawal threshold (MWT) was measured on 1 day before establishing the model and 1-7 days after establishing the model.The spinal dorsal horns of lumbar enlargement segments were removed on day 7 after establishing the model for determination of the expression of glial fibrillary acidic protein (GFAP) (by Western blot) and co-expression of G-protein-coupled receptor 18 with GFAP (by double-label immunofluorescence).Results Compared with group S,the MWT was significantly decreased at 1-7 days after establishing the model,and the expression of GFAP was up-regulated in group P (P＜0.05).Compared with group P,the MWT was significantly increased at 3-7 days after establishing the model,and the expression of GFAP was down-regulated in group R (P＜0.05).G-protein-coupled receptor 18 was co-expressed with GFAP.Conclusion Exogenous resolvin D2 can reduce radicular pain in a rat model of non-compressive lumbar intervertebral disc herniation,and the mechanism is related to inhibiting activation of astrocytes in the spinal dorsal horns.
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Objective To evaluate the efficacy of facial acupoint injection combined with nerve block in treating hemifacial spasm. Methods Fifty?four patients of both sexes, aged 26-73 yr, with the course of disease 6 months-17 yr from February 2015 to January 2016, were included in this study accord?ing to diagnostic criteria for hemifacial spasm based on expert consensus in 2014. Patients were divided into 2 groups(n=27 each)using a random number table: facial acupoint injection combined with nerve block group(group A)and nerve block group(group B). In group A, the mixed injection 2 ml(2% lidocaine hydrochloride 5 ml, compound betamethasone 1 ml, mecobalamin 1 mg, diluted to 18 ml in normal sa?line)was given at facial Sibai, Jiachengjiang, Xiaguan and Yifeng acupoints once every 5 days, 3 times in total;facial nerve block was performed via the stylomastoid foramen with the mixed injection(5 ml) previously described once every 5 days, 3 times in total;stellate ganglion block was performed on the right and left side alternately once a day, 5 times on each side. In group B, patients received facial nerve block and stellate ganglion block, and no facial acupoint injection was applied. The Cohen rating and Shorr out?come were recorded before treatment and at 10th day, 3rd month and 1 yr after treatment, and the recur?rence was recorded at 3 months and 1 yr after treatment. Results Compared with group B, Cohen classifi?cation was significantly decreased, and the effective rate was increased at 10th day, 3rd month and 1 yr af?ter treatment, and the recurrence rate was decreased at 3 months and 1 yr after treatment in group A(P<0.05). Conclusion The facial acupoint injection combined with nerve block produces better long?term ef?ficacy in treating hemifacial spasm than nerve block alone.
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Objective To explore the effect of resolvin D2 ( RvD2) on radicular pain induced by interver-tebral disc herniation. Methods Thirty-six male Sprague-Dawley rats were randomly divided into a sham opera-tion group, a model group and an RvD2 group, each of 12. Non-compressive lumbar disc herniation was induced in the rats in the model and RvD2 groups using the autologous nucleus pulposus filling method. Those in the sham group had the surgical site exposed without any other treatment. After the modeling, 10μl of phosphate-buffered sa-line solution was administered intrathecally to the rats in the sham and model groups for 3 days, while the rats in the RvD2 group received 10 ng/10 μl of RvD2 intrathecally as well. Paw withdrawal thresholds (50％PWT) were ob-served 1 day before modeling and 7 days afterward for the rats of all three groups. On the 7th day after modeling, the L4 to L6 spinal dorsal horns on the surgery side were resected to detect the protein expression of phosphorylated protein kinase B ( p-AKT) , protein kinase B ( t-AKT) , phosphorylated glycogen synthase kinase 3β( p-GSK-3β) and glycogen synthase kinase 3β( GSK-3β) using western blotting. The protein levels of tumor necrosis factor alpha ( TNF-α) , interleukin-6 ( IL-6) , transforming growth factor-β1 ( TGF-β1) and interleukin-10 ( IL-10) were de-termined using enzyme-linked immunosorbent assays. Results On the 1st and 7th day after modeling, significant differences were observed between the model and sham groups in terms of the 50％PWT. From the 3rd day the aver-age 50％PWT in the RvD2 group was significantly higher than that of the model group at the same time points. On the 7th day after the modeling the average p-AKT and p-GSK-3βprotein levels of the model and RvD2 groups were significantly different from that of the sham group, and the model group′s average was also of significantly different from that of the RvD2 group. The average protein levels of the pro-inflammatory cytokines TNF-αand IL-6, as well as of the anti-inflammatory factors TGF-β1 and IL-10 in the dorsal horns of the model group and the RvD2 group were also significantly different on the 7th day, and both were significantly different from the sham group′s average. Conclusion RvD2 can alleviate radicular pain in rats with non-compressive disc herniation. The mechanisms might involve inhibition of GSK-3β activity, down-regulation of pro-inflammatory factors and up-regulation of anti-inflammatory factors.
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Objective To investigate the analgesic effect of Resolvin D1 (RvD1) on radicular pain induced by herniated nucleus pulposusand its underlying mechanism.Methods Fifty-six male rats were randomly divided intoa sham group,a model group,a 10 ng group anda 100 ng group,each of 14.The rat model of non-compressive lumber disc herniation was established in all except the sham group.The former two groups were then injected with 10 μl of phosphate buffer solution (PBS) while the latter 2 groups were injected with 10 μl of PBS containing 10 and 100 ng of RvD1 respectively daily for three successive days.The rats' 50％ paw withdrawal threshold (PWT) was evaluated 1 day before and on 7 successive days after surgery.On day 7 the rats' spinal cords were removed to assess the expression of tumor necrosis factor-or (TNF-α),interlukin-1β (IL-1β) and interlukin-10 (IL-10) using ELISA methods.The levels of ERK and NF-κB/p65 were measured using Western blotting.Results Theaverage 50％PWT of the model group decreased significantly from day 1 to day 7 compared with the sham group,but was significantly lower thanthe RvD1 10 ng group from day 3 to day 7.Moreover the 50％PWT in the RvD1 100 ng group increased significantlyfrom day 2 to day 7 compared with the model group (P＜0.05).The average expression of both TNF-α and IL-1β of the model group was upregulated significantly and that of IL-10 decreased significantly compared with the sham group.Compared with the model group,the expression of TNF-α and IL-1β decreased significantly (P＜0.05)and the level of IL-10 was significantlyup-regulated (P＜0.05) both in the RvD1 10 ng group and 100 ng group.Moreover,the changes were larger in the RvD1 100 ng group (P＜0.05).Compared with the sham group,the levels of p-ERK and NF-κB/p65 in the model group were significantly up-regulated (P＜0.05).Compared with the model group,intrathecal injection of RvD1 (10 ng or 100 ng) significantly decreased the expressions of p-ERK and NF-κB/p65 (P＜0.05).Moreover,the decrease wasgreater in the RvD1 100 ng group (P＜0.05).Conclusions RvD1 might alleviate the radicular inflammation and pain byregulating the balance of inflammatory mediators and activation of p-ERK and NF-κB/p65 pathways.It may offer novel therapeutic approaches for the management of lumbar disc herniation.
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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.
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Objective To investigate the effect of lipoxin A4 LXA4) on radicular pain caused by intervertebral disc herniation.Methods Non-compressive intervertebral disc herniation was induced into forty-eight adult male Sprague-Dawley rats,and they were divided into a sham group (sham operation + 10 μl normal saline),a control group (modeled + 10 μl normal saline),an LXA4 10 ng group (modeled + 10 ng LXA4) and an LXA4 100 ng group (modeled + 100 ng LXA4),with 12 rats in each group.The normal saline (10 μl) or LXA4 (10 μl) was administered intrathecally right after the operation and on each of the three succeeding days.General behavior was observed and the 50％ paw withdrawal threshold (50％ PWT) was measured.On postoperative day 7 all the rats were killed and the ipsilateral lumbar (L4~) segments of their spinal dorsal horns were removed for determination of the expression of p-JNK,t-JNK,p-ERK and t-ERK proteins using western blotting.TNF-α,IL-1β and TGF-β1 expression were determined using ELISA.Results There was no significant difference in the 50％PWT of the sham group before and after surgery,but the 50％ PWTs of the control group and the LXA4 10 ng group were significantly decreased after the operation compared with their values beforehand and significantly lower than the value of the sham group at all time points.Moreover,the 50％ PWT of the LXA4 10 ng group on postoperative days 3 and 5 was significantly higher than the control group;as was the value of the LXA4 100 ng group on postoperative days 2,3,4,5,6 and 7.The p-JNK and p-ERK expression in the control group,the LXA4 10 ng group and the LXA4 100 ng group were all increased significantly more than in the sham group,but their expression in the LXA4 10 ng group and LXA4 100 ng group were decreased significantly more in a dose-dependent manner compared with the control group,with the LXA4 100 ng group showing the greatest decrease.There were no significant differences in t-JNK or t-ERK expression within each group.Conclusion LXA4 can alleviate radicular pain caused by non-compressive lumbar intervertebral disc herniation.The underlying mechanism involves inhibiting the activation of the ERK and JNK pathways,reducing the expression of pro-inflammatory cytokines and increasing the expression of anti-inflammatory cytokines.
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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.
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Objective To investigate the alteration of nuclear factor kappa B( NF-κB) and tumor necrosis factor-a (TNF-α) mRNA and protein in hippocampus in chronic constrictive injury (CCI) model of rats. Methods Seventy-six male Wistar rats were randomly divided into 2 groups ( n = 38): the CCI group which received the chronic constriction injury and the sham group which received the sham operation as control. The mechanical and thermal nociceptive thresholds were assessed with paw withdrawal latency (PWL) to von Frey filaments and radiant heat at 1d before and ld,4d,7d,14d and 28d after CCI operation. Five animals were sacrificed at each time point for real-time polymerase chain reaction (real-time PCR) and another three animals sacrificed at 7d postoperation for immunofluorescence histochemical staining. Results The thresholds to mechanical and thermal stimuli decreased obviously after operation in CCI group. The expressions of TNF-α and NF-κB mRNA began to increase at ld( (2.079 ±0. 104)times and 4d( ( 1.640 ± 0.064) times) after operation and reached the peak at 7d ((2.748 ±0.147)times, (2.010 ±0.096)times) ,then the expressions of TNF-a mRNA began to decrease,while the expressions of NF-kB mRNA maintained at a high level throughout the experiment. The result of immunofluorescence histochemical staining revealed that NF-kB and TNF-α protein expressions at 7 day increased significantly on the hippocampus,which was consisted with NF-κB and TNF-a mRNA levels. Conclusion The activation NF-κB and TNF-α in hippocampus may be involved in the procession of neuropathic pain.
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Objective To investigate the expression of nuclear factor kappa B(NF-Κb)in the lumbar spinal cord in chronic constrictive injury(CCI)of the sciatic nerve.Methods Foay-eight male Sprague-Dawley rats were divided into 2 groups(n=24 in each).The operation group received CCI of the sciatic nerve and the sham-operation group received a sham operation as a control.Their mechanical and thermal nociceptive thresholds were assessed with paw withdrawal latency (PWL) using von Frey filaments and radiant heating.Four rats were sacrificed at each time point and segments of their lumbar spinal cords were examined using Western blotting.Results Mechanical and thermal stimulation thresholds were reduced significantly after the operation.Expression of NF-Κb protein increased significantly in the ipsilateral dorsal horn of the spine compared with the contralateral side and compared with the sham-operation group.NF-Κb protein expression began to increase on the 1 st day after the operation,reached a peak at the 7th day,and stayed high throughout the experiment.Conclusion NF-Κb in the dorsal horn of the spine may be involved in neuropathie pain after CCI.
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Objective To investigate the change in NF-κB activity in astrocytes in spinal dorsal horn in a rat model of neuropathic pain and the underlying mechanism. Methods Sixteen male SD rats aged 2-3 months weighing 220-280 g were randomly divided into 2 groups ( n = 8 each) : sham operation group (group S) and CCI group. Neuropathic pain was induced by chronic constrictive injury (CCI) . Right sciatic nerve was exposed and 4 loose ligatures were placed on the sciatic nerve at 1 mm intervals with 4-0 chromic catgut. In group S the right sciatic nerve was exposed but not ligated. The paw withdrawal threshold (PWT) to von Frey filament stimulation and paw withdrawal latency (PWL) to radiant heat stimulation were measured at 1 d before (baseline) and 7 d after operation. The animals were then killed and the lumbar segment of the spinal cord (L_(4-6)) was removed. The expression of NF-κB in the astrocytes in spinal dorsal horn was determined by immuno-histochemistry. Results PWT and PWL to mechanical and thermal stimuli were significantly decreased after operation as compared with the baseline before operation in group CCI. The number of NF-κBp65 immunoreaction positive cells in the spinal dorsal horn on the operated side was significantly larger in group CCI than in group S. Conclusion NF-κB signal transduction pathway in the astrocytes in the spinal dorsal hom may be involved in neuropathic pain.
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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.
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Objective To investigate the toxic effects of ozone on rat astrocytes in vitro. Methods Twelve 1-2 day old Wistar rats were anesthetized with ether. Their brains were removed. Astrecytes were enzymatically isolated with trypsin and cultured. After different-speed cultivation and serial passage, astrocytes were seeded into 24 well plates and divided into 4 groups (n = 7 each): group Ⅰ normal control (group C) and group Ⅱ , Ⅲ , Ⅳ (group O2-O340, 60, 80) were exposed to ozone 40, 60 and 80 μg/ml respectively. After 2 and 4 h incubation, the astroeytes were examined under microscope. Supernatant was collected and cells were broken for determination of SOD activity and MDA content in the cells and LDH leaking ratio. Results Cells in group O2-O3 60 and 80 exhibited hypertrophy, vacuoles and black denatured grains in the cytoplasm and increase in proceses. SOD activity and MDA content were significantly increased at the end of 2 h incubation in group O2-O340, 60 and 80 as compared with blank control group. LDH leaking ratio and MDA content were increased in a concentration dependent manner at the end of 2 h and 4 h incubation, while SOD activity was decreased concentration dependently at the end of 4 h incubation. SOD activity was signifieandy decreased, while LDH leaking ratio was increased at 4 h as compared with that at 2 h in group O2-O360 and 80. Conclusion Ozone has toxic effects on cultured rat astroeytns in a concentration and time dependent manner.
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Objective To evaluate the analgesic effect and safety of subarachnoid chromaffin cell allograft for terminal cancer pain Methods Ten patients with intractable cancer pain despite traditional treatments were randomly divided into two groups In test group(n=4), 2ml of the suspension chromaffin cells cultured in vitro for 3 days was injected into the subarachnoid space through lumber puncture The same amount of cell free culture solution was injected intrathecally in control group(n=6) Opioids were administered continuously after transplantation The intensity of pain was assessed by VAS, the dose of opioids taken was recored,and the catecholamine and enkephalin concentrations in cerebrospinal fluid and immune function were measured before and after transplantation Results The VAS scores declined markedly in both groups after transplantation (P
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The new approaches were designed to have anti-inflammatory analgetics act concentratively on the surrounding of pathological nerve roots or to inject collagenase directly to the protruding lumbar intervertebral disc for more simple and effective treatment of the low back pain. Method: The approaches to epidural lateral recess via incisura margo lateralis of lamina arcus vertebrae or margo medialis of joint processus artieularis were supposed and applied in clinic after study of anatomy and analysis of lumbar vertebra X-ray and CT photographes. The safety and feasibility of the approaches were evaluated according to the test of block region, measurement of therapeutic effect and monitoring of CT during the whole procedures in some cases. Result: A total of 170 cases via the incisura margo lateralis of lamina areus vertebrae were tested, 168 cases turned out to be successful and 2 cases failing. Another 15 cases via margo medialis of joint proeessus articularis, all punctures were successful. Conclusion: The approaches can lead to epidural lateral recess precisely to obtain a better treatment. When the technique is applied to discolysis, no X-ray machine will be needed and therefore no injury of X-ray occurs to both doctors and patients.
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The distance between the medial border of the two articular processes was measured by CT and MRI. At the level of C5-6-7 it was 23-25 mm(24mm on average). If the distance is 24 mm at C5-6 a tuohy needle is inserted 10 mm(24?2-2) lateral to the midline. When the needle touches vertebral lamina, withdraw the needle 2mm and redirect it cephalad at 45-60, Resistance is encountered when the needle touches ligament flavum. When the resistance disappears, the tip of the needle has entered the lateral recess via tuohy needle. This approach was used in 45 patients with satisfactory results. In ten patients the position of the cathter was confirmed with contrast medium by CT and MRI.