Antigens, Neoplasm ; metabolism ; Antigens, Surface ; metabolism ; Colorectal Neoplasms ; immunology ; pathology ; Humans ; Prognosis ; Trophoblasts ; immunology

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 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.

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.

The axon initial segment (AIS) is a specialized structure that controls neuronal excitability via action potential (AP) generation. Currently, AIS plasticity with regard to changes in length and location in response to neural activity has been extensively investigated, but how AIS diameter is regulated remains elusive. Here we report that COUP-TFI (chicken ovalbumin upstream promotor-transcription factor 1) is an essential regulator of AIS diameter in both developing and adult mouse neocortex. Either embryonic or adult ablation of COUP-TFI results in reduced AIS diameter and impaired AP generation. Although COUP-TFI ablations in sparse single neurons and in populations of neurons have similar impacts on AIS diameter and AP generation, they strengthen and weaken, respectively, the receiving spontaneous network in mutant neurons. In contrast, overexpression of COUP-TFI in sparse single neurons increases the AIS diameter and facilitates AP generation, but decreases the receiving spontaneous network. Our findings demonstrate that COUP-TFI is indispensable for both the expansion and maintenance of AIS diameter and that AIS diameter fine-tunes action potential generation and synaptic inputs in mammalian cortical neurons.
Action Potentials ; Animals ; Axon Initial Segment ; COUP Transcription Factor I ; DNA-Binding Proteins/physiology* ; Mammals ; Mice ; Transcription Factors

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.

Objective To evaluate the molluscicidal effects and costs of spraying 20% suspension concentrate of pyricloben-zuron sulphate (SCPS) with drones against Pomacea canaliculata in paddy environments, so as to provide insights into the extensive applications of pyriclobenzuron against P. canaliculata. Methods On July 2022, a paddy field was selected from Nanchang City, Jiangxi Province as the study area, and 72 independent rectangular plots measuring 2 m × 1 m were allocated in the study area, with 1 m interval between each plot, and 20 P. canaliculata snails gently placed in each plot. The activity of 25% wettable powder of pyriclobenzuron sulphate (WPPS) by manual spraying at doses of 0.50, 1.00, 2.00 g/m² and 4.00 g/m² against P. canaliculata was tested in 54 plots, and manual spraying of 50% wettable powder of niclosamide ethanolamine salt (WPNES) at a dose of 0.10 g/m² served as a chemical control, while manual spraying of the same volume of clean water served as a blank control, with 9 plots in each group. The activity of SCPS against P. canaliculata was tested in the remaining 18 plots. Based on the molluscicidal tests of WPPS, the molluscicidal effect of SCPS by manual spraying at doses of 0.20, 0.30, 0.40 g/m² and 0.50 g/m² against P. canaliculata was evaluated, and manual spraying of WPNES at a dose of 0.10 g/m² served as a chemical control, while manual spraying of the same volume of clean water served as a blank control, with three plots in each group. On July 2023, 14 paddy fields with a mean living P. canaliculata density of > 5 snails/m² were selected from Yujiang District, Yingtan City, Jiangxi Province for molluscicidal tests. Based on the molluscicidal effect of pyriclobenzuron against P. canaliculata in plots, the molluscicidal effects of WPPS by manual spraying at doses of 0.25, 0.50 g/m² and 1.00 g/m² and manual applications of WPPS at dose of 0.25, 0.50, 1.00 g/m² and 2.00 g/m² mixed with soil were tested, and manual spraying of 0.10 g/m² WPNES served as a chemical control group, while manual spraying of the same volume of clean water served as a blank control, with one paddy field in each group. Based on the effect of pyriclobenzuron against P. canaliculata in plots, the activity of SCPS sprayed with drones at doses of 0.25 g/m² and 0.50 g/m² mixed in water at 2 kg/667 m² and 4 kg/667 m² was tested against P. canaliculata, and spraying of the same volume of clean water with drones served as a blank control. All P. canaliculata snails were captured 3 days and 7 days following chemical treatment in plots and paddy fields and identified for survival, and the mortality and corrected mortality of P. canaliculata snails were estimated. In addition, the areas of chemical treatment, amount of molluscicide use and labor costs of chemical treatment were estimated in molluscicidal tests in paddy fields, and the costs of chemical treatment for an area covering 667 m² by drones and manual applications were calculated. Results The mortality of P. canaliculata snails was all 100% in plots 3 days and 7 days following spraying WPPS at doses of 0.50, 1.00, 2.00 g/m² and 4.00 g/m², and the mortality rates of P. canaliculata snails were 66.67% to 100.00% 3 days post-treatment with SCPS at various doses (χ² = 277.897, P < 0.05) and 76.67% to 100.00% 7 days post-treatment (χ² = 274.206, P < 0.05). The mortality rates of P. canaliculata snails were 98.19% to 100.00% 3 days post-treatment with WPPS at various doses in paddy fields. There was a significant difference in the mortality of P. canaliculata snails among WPPS treatment groups and controls (χ² = 270.778, P < 0.05), and there were no significant differences between WPPS treatment groups and the chemical control group (all P values > 0.05), while there were significant differences in the mortality of P. canaliculata snails between WPPS treatment groups and the blank control group (all P values < 0.05). The mortality rates of P. canaliculata snails were 89.83% to 95.31% 3 days post-treatment with SCPS at various doses sprayed with drones, and there was a significant difference in the mortality of P. canaliculata snails among SCPS treatment groups and the blank control group (χ² = 1 132.892, P < 0.05). There were no significant differences in the mortality of P. canaliculata snails among SCPS treatment groups or water mixture groups (all P values > 0.05), and there were significant differences in the mortality of P. canaliculata snails between SCPS treatment groups and the blank control group (all P values < 0.05). The mortality rates of P. canaliculata snails were 94.62% to 100.00% 7 days post-treatment with SCPS at various doses sprayed with drones, and there was a significant difference in the mortality of P. canaliculata snails among SCPS treatment groups and the blank control group (χ² = 1 266.932, P < 0.05), with the highest mortality found following spraying 0.50 g/m² SCPS mixed in 2 kg/667 m² water with drones (P < 0.05). The costs of P. canaliculata snail control by drones and manually were 35.85 Yuan/667 m² and 43.33 Yuan/667 m²; however, the snail control efficiency was 6.67 times higher by drones than by manual applications. Conclusions SCPS sprayed with drones is highly active against P. canaliculata snails in paddy fields. SCPS sprayed with drones is highly efficient and low in cost for P. canaliculata snail control in paddy fields, beaches and river courses.