目的观察分期综合疗法治疗颈椎间盘突出症的疗效。方法将100例颈椎间盘突出症住院患者随机分为治疗组(采用分期综合疗法)与对照组(采用传统综合疗法)各50例,评定两组患者的疗效。结果治疗组总有效率88%,对照组总有效率78%,两组间有显著性差异(P<0.05)。结论分期综合疗法治疗颈椎间盘突出症的疗效优于传统综合疗法。

Objective To observe the effects of electro-acupuncture (EA) at “Weizhong” (BL40) on expressions of Fas and FasL in rat models with lumbar disc degeneration; To discuss its mechanism of action.MethodsThirty SD rats were randomly divided into sham-operationgroup, model group and EA group, with 10 rats in each group. The sham-operation group was treated with sham operation to incide local skin; the model and EA groups established rat model of lumbar disc degeneration by puncturing the annulus fibrosus. Four weeks after modeling, rats in the sham-operation and model groups received fixed treatment under identical condition, and rats in the EA group were treated with EA at “Weizhong” (BL40) in 20 minutes, once a day, for 4 weeks. After treatment, Western blot and RT-PCR technology were used to test the protein and mRNA expression levels of Fas and FasL.ResultsThe expressions of Fas and FasL in the model group were higher than those in the sham-operation group (P<0.05); protein and mRNA expression levels of Fas and FasL in the EA group were lower than those in the model group (P<0.05). ConclusionEA at “Weizhong” (BL40) can reduce the expressions of Fas and FasL to inhibit the development of lumbar disc degeneration.

OBJECTIVE: To investigate the protective effects of fibronectin (Fn) on vascular endothelial cells. METHODS: Block reticuloendothelial system model compound with injury of tail vein and femur fractures in rats were used. The level of plasma Fn and surface Fn of peritoneal macrophage cells, the numbers of circulating endothelial cells and the injured tail vein were observed. RESULTS: Fn increased the level of plasma Fn and peritoneal macrophage cells surface Fn,but decreased the number of circulating endothelial cells after infusing Fn 1.7 mg/10 g before injury. Under the scanning electron microscope, only a local focus injury to vascular endothelial cells in the rats was observed after Fn was infused, but a piecemeal injury and detachment of vascular endothelial cells were observed in rats without giving Fn. CONCLUSIONS: Fn has obvious protective effects on vascular endothelial cells in rats.

Objective: To investigate the analgesic mechanism of Tuina (Chinese therapeutic massage) by observing the effect of the N-methyl-D-aspartate receptor subunit 2B (NR2B)/postsynaptic density-95 (PSD-95) pathway on the dendritic structure of spinal cord dorsal horn in rats with lumbar disc herniation. Methods: Fifty Sprague-Dawley rats were randomly divided into a blank group, a model group, a Tuina group, a blocker agent group, and a blocker agent + Tuina group. The sciatic nerve chronic constriction injury (CCI) model was prepared by the sciatic nerve ligation method. From the 4th day after modeling, rats in the Tuina group and the blocker agent + Tuina group were subject to daily Tuina intervention, and those in the blocker agent group and the blocker agent + Tuina group were daily intrathecally injected with NR2B blocker agent (MK-801). The spontaneous pain score was used to observe the pain behavior of all rats. The expression levels of NR2B and downstream PSD-95 were measured by immunohistochemistry, and the dendritic structure changes were observed by Golgi staining for rat spinal cord dorsal horn after 14 d of continuous intervention. Results: Compared with the blank group, the degree of rat spontaneous pain after CCI was elevated in both the model and the Tuina groups (P<0.01) and was reduced in the Tuina group after the Tuina intervention compared with the model group (P<0.05). Compared with the model group, the rat spontaneous pain level after blocking NR2B was reduced in both the blocker agent group and the blocker agent + Tuina group (P<0.05). The NR2B and PSD-95 protein levels were significantly higher in the model group compared with the blank group (P<0.01); the total number of dendritic branches was increased (P<0.01), and the total dendritic length became longer (P<0.01) in the spinal cord dorsal horn. The rat NR2B and PSD-95 protein levels were significantly decreased in the Tuina group compared with the model group (P<0.01); the total dendritic branch number was reduced (P<0.01) and the total length was shortened (P<0.01) in the spinal cord dorsal horn. After blocking NR2B, the expression levels of NR2B and downstream PSD-95 protein were significantly lower in both the blocker agent group and the blocker agent + Tuina group compared to the model group (P<0.01). The total branch number was significantly reduced (P<0.01), and the total length was significantly shortened (P<0.01) of the dendrites in the spinal cord dorsal horn. Conclusion: Tuina may exert an analgesic effect by remodeling the dendritic structure in the spinal cord dorsal horn in rats with lumbar disc herniation, and its mechanism may be related to the inhibition of NR2B/PSD-95 signaling pathway.

Objective To investigate the effects of abdominal tuina on the expression of PI3K and N-methyl-D-aspartate receptor(NMDAR)subunit NR1 in spinal dorsal horn and the morphology of spinal dorsal horn neurons in ulcerative colitis(UC)rats;To explore its mechanism of action in treating UC.Methods Totally 36 SD rats were randomly divided into normal group,model group,abdominal tuina group,mesalazine group,PI3K stimulation group and PI3K stimulation + abdominal tuina group,with 6 rats in each group.The UC model in rats was simulated by drinking dextran sulfate solution freely.The abdominal tuina group and the PI3K stimulation + abdominal tuina group were given abdominal tuina intervention,the mesalazine group was given mesalazine solution for gavage,and the PI3K stimulation group and PI3K stimulation + abdominal tuina group were given intrathecal injection of PI3K agonist,once a day,for consecutive 15 days.Abdominal withdrawal reflex(AWR)score and acetic acid twist were used to observe the abdominal pain symptoms in rats.The expression of PI3K and NR1 in spinal dorsal horn were detected by immunofluorescence staining and Western blot,and the morphological changes of spinal dorsal horn neurons were observed by Nissl staining.Results Compared with the normal group,AWR score and twisting times of rats in model group significantly increased(P<0.01),the expression of PI3K and NR1 protein in spinal dorsal horn significantly increased(P<0.05,P<0.01),the morphology of spinal dorsal horn neurons was disordered,forming a large number of vacuolar like structures,and the Nissl body structure was fuzzy and incomplete.Compared with the model group,AWR scores and twisting times of abdominal tuina group and mesalazine group significantly decreased(P<0.05,P<0.01),and the expression of PI3K and NR1 protein significantly decreased(P<0.05,P<0.01),the edema of spinal dorsal horn neurons was milder,with fewer vacuolar changes and an increase in the number of Nissl bodies;AWR scores and twisting times of PI3K stimulation group and PI3K stimulation + abdominal tuina group significantly increased(P<0.05,P<0.01),and the expressions of PI3K and NR1 protein increased(P<0.05,P<0.01),a large number of neurons underwent pyknosis and necrosis,and the number of Nissl bodies decreased,even dissolving and disappearing.Conclusion Abdominal tuina can effectively improve the symptoms of abdominal pain in UC model rats,and its mechanism may be related to inhibiting the expression of PI3K and NR1 in spinal dorsal horn and improving the morphology of spinal dorsal horn neurons.

Objective To observe the effects of tuina on P2Y12/RhoA/ROCK2 pathway and c-Fos protein expression of microglia in spinal dorsal horn of sciatic nerve chronic compressive injury(CCI)rats;To explore the mechanism of tuina in treating lumbar disc herniation.Methods A CCI model of right sciatic nerve was used to simulate neuropathic pain in lumbar disc herniation.24 male SD rats were randomly divided into blank group,model group and tuina group,with 8 rats in each group.After 4 days of modeling,massage intervention was used to in the tuina group for 14 days.The paw withdrawal threshold(PWT)and paw withdrawal latency(PWL)of rats before and on the 4th,10th,and 17th day after modeling were observed,immunofluorescence staining was used to detect the expression of Iba1 and P2Y12 protein in right spinal dorsal horn,Western blot was used to detect the expressions of RhoA and ROCK2 protein in right spinal dorsal horn,immunohistochemistry staining was used to detect the number of c-Fos positive cells in right spinal dorsal horn.Results Compared with the blank group,the PWT and PWL of the model group were significantly reduced on the 4th,10th,and 17th day after modeling(P<0.001),while the expressions of Iba1,P2Y12,RhoA and ROCK2 protein in right spinal dorsal horn significantly increased(P<0.001,P<0.05),the number of c-Fos positive cells significantly increased(P<0.001).Compared with the model group,the PWT and PWL of the tuina group significantly increased on the 10th and 17th day after modeling(P<0.05,P<0.01,P<0.001),and the expressions of Iba1,P2Y12,RhoA and ROCK2 protein in right spinal dorsal horn significantly decreased(P<0.001,P<0.01,P<0.05),and the number of c-Fos positive cells significantly reduced(P<0.001).Conclusion Tuina may inhibit the activation of microglia by regulating P2Y12/RhoA/ROCK2 pathway and c-Fos protein expression in spinal dorsal horn,reduce neuronal excitability,and exert analgesic effects on lumbar disc herniation.

Objective To observe the effect of tuina on glutamate uptake and synaptic cleft in the spinal dorsal horn of rats with neuropathic pain through astrocyte NDRG2/GLT-1 pathway,and to explore the potential analgesic mechanism of tuina on neuropathic pain.Methods A total of 54 SD rats were randomly divided into naive group,model group and tuina group(n=18).The CCI model was established in the model group,and the tuina group was treated with acupressure at"Weizhong"(BL 40)from the 4th day after CCI model was successfully established for 14 days.The changes of paw withdrawal threshold at different time points were observed to evaluate the analgesic effect of tuina.Immunofluorescence was used to observe the co-expression of NDRG2?GLT-1 and astrocytes in the spinal dorsal horn.The mRNA levels of NDRG2 and GLT-1 in astrocytes were detected by quantitative real time polymerase chain reaction.The concentrations of glutamate in synaptic cleft were measured by liquid chromatography coupled to tandem mass spectrometry.The width of the synaptic cleft was observed by transmission electron microscopy.Results Compared with the naive group,the paw withdrawal threshold in the CCI group decreased continuously(P<0.01).The number of NDRG2 and GFAP co-labeling positive cells in the spinal dorsal horn increased significantly(P<0.01),and the number of GLT-1 and GFAP co-labeled positive cells was significantly reduced(P<0.01).NDRG2 mRNA expression was up-regulated and GLT-1 mRNA expression decreased in astrocytes(P<0.01).The concentrations of glutamate increased significantly(P<0.01);The synaptic cleft was significantly narrowed(P<0.05).After tuina intervention,the above trend was significantly reversed.Compared with the model group,the paw withdrawal threshold of the tuina group increased from 11 days after CCI(P<0.01),the number of NDRG2 and GFAP co-labeling positive cells in the spinal dorsal horn was significantly reduced(P<0.01),and the number of GLT-1 and GFAP co-labeled positive cells increased significantly(P<0.01);down-regulated the expression of NDRG2 mRNA and restored GLT-1 mRNA expression in astrocytes(P<0.01);decreased glutamate concentration in synaptic cleft(P<0.05),the synaptic cleft was relatively widened(P<0.05).Conclusion Tuina alleviates pain in CCI rats at the spinal cord level possibly by promoting the uptake of glutamate by NDRG2/GLT-1 pathway in astrocytes,restoring the width of synaptic cleft and reversing synaptic plasticity.

ObjectiveTo explore the possible mechanism of Tuina at Weizhong （BL 40） for relieving sciatica from the perspective of hippocampal synaptic plasticity. MethodsSixty SD rats were randomly divided into sham operation group， model group， Tuina group， MK-801 group， MK-801 plus Tuina group， 12 rats in each group. After lateral ventricular cannulation， rats model with chronic compression injury of the right sciatic nerve were prepared in all groups except the sham operation group. On day 4 after modelling， rats in the Tuina group start Tuina at Weizhong （BL 40） for 10 mins once a day for a total of 14 days； rats in the MK-801 group started injecting with 0.25 μg/μl of the N-methyl-D-aspartate receptor 2B （NR2B） blocker， dizocycline （MK-801）， 0.5 μl of which was administered daily in the lateral ventricle for 14 days. Rats in the MK-801 plus Tuina group underwent Tuina after 30 mins when completing MK-801 injection in the lateral ventricle， in the same way as above； rats in the model group and the sham operation group did not undergo any intervention. Spontaneous pain behaviour scores and paw withdraw thresholds （PWTs） were examined on day 1 （base value） before modelling and on day 4， 10， 14 and 18 after modelling； and on day 19， the brain tissues of the rats in each group were sampled and the number and morphology of the Nysted-positive cells in the hippocampal CA3 region were observed using Nysted staining； and the number of synapses， the thickness of postsynaptic dense material， the length of active band and the curvature of synaptic interface in hippocampal CA3 region were observed by transmission electron microscopy； and the expression of synapse-associated proteins NR2B and postsynaptic density protein-95 （PSD95） in hippocampal CA3 was detected by immunofluorescence staining and immunoblotting. ResultsCompared with the same time in the sham operation group， spontaneous pain scores significantly increased and PWTs decreased on day 4， 10， 14， and 18 after modelling in the model group （P＜0.05）； compared with the model group， spontaneous pain scores in Tuina group of rats significantly decreased on day 10， 14， and 18 after modelling， and PWTs significantly increased on day 14 and 18 after modelling （P＜0.05）. Compared with Tuina group， spontaneous pain scores increased on day 10， 14， and 18 of modelling， and PWTs decreased at days 14 and 18 of modelling in the MK-801 plus Tuina group had higher spontaneous pain scores on days 10， 14， and 18 after modelling and lower PWTs on days 14 and 18 after modelling （P＜0.05）. Compared with the sham operation group， the neuronal arrangement in the hippocampal CA3 region of the rats in the model group was disordered， with decreased number of Nysted-positive cells and synapses， reduced thickness of postsynaptic densities， length of active bands， and curvature of synaptic interfaces， wider synaptic gaps， and decreased immunofluorescent positive expression of NR2B and PSD95 as well as the expression of immunoblotting proteins in hippocampal CA3 region （P＜0.05）. Compared with the model group， more dense arranged nerve cells， the number of Nysted-positive cells， the number of synapses， the thickness of postsynaptic dense material， the length of active bands increased， the synaptic gap became significantly narrower， and the positive expression of immunofluorescence and immunoblotting protein expression of NR2B， PSD95 increased in the rat hippocampal CA3 region of Tuina group （P＜0.05）. Compared with Tuina group， the neuronal morphology of the hippocampal CA3 region in MK-801 plus Tuina group was severely damaged， and the number of Nystrom's-positive cells， the number of synapses， the thickness of post-synaptic densities， the length of active bands， and the curvature of synaptic interfaces reduced， the synaptic gaps became wider， and the immunofluorescent positive expression of NR2B， PSD95， and the expression of immunostained proteins decreased （P＜0.05）. ConclusionTuina at "Weizhong" （BL 40） showed significant analgesic effect， and one of the possible mechanisms concluded as significantly increasing the levels of NR2B and PSD95 protein expression in hippocampal CA3 region and thus modulating the synaptic plasticity of the hippocampus.