RESUMO
ObjectiveTo explore the possible mechanism of Tongdu Xingshen needling method （通督醒神针刺法） on post-stroke cognitive impairment. MethodsSD rats were randomly divided into a normal group （n=12）， a sham surgery group （n=12）， a model group （n=12）， and a electroacupuncture group （n=13）. The rats in the model group and electroacupuncture group were subjected to the wire bolus method to establish the rats model with learning memory impairment after cerebral ischaemia-reperfusion. After successful modelling， the rats in the electroacupuncture group were given electroacupuncture interventions at “Shenting （GV 24）” and “Baihui （GV 20）” once a day for 30 minutes for 14 days. The other three groups did not receive other interventions but grasp. A 5-day localisation navigation experiment was conducted on the 9th day of intervention， and a spatial exploration experiment was conducted on the 14th day of intervention to evaluate the learning and memory abilities of the rats. After the spatial exploration experiment， hippocampal tissues were taken from each group of rats， and the changes in the volume of cerebral infarction were observed by TTC staining； the changes in the morphology of pyramidal neurons and the density of dendritic spines in the CA1 area of the hippocampus were observed by Golgi staining； protein immunoblotting was used to detect the relative protein expression of the subunits of the α-amino-3-carboxy-5-methylisoxazole-4-propionic acid （AMPA） receptor including glutamate receptor 1 （GluR1）， glutamate receptor 2 （GluR2）， glutamate receptor 3 （GluR3） and auxiliary proteins TARPγ2， TARPγ8 in hippocampal tissues of rats in each group； the real-time fluorescence quantitative PCR was used to detect GluR1， GluR2， GluR3 mRNA levels in the hippocampal tissues of rats. ResultsIn the localisation navigation experiment， compared with the normal group and sham surgery group， the escape latency and total distance of rats in the model group were significantly extended （P＜0.05） at day 1， 2， 3， 4， and 5； and the escape latency and total distance of rats in the electroacupuncture group tended to be significantly shorter than those in the model group （P＜0.05）. In the spatial exploration experiment， compared with the normal group and the sham surgery group， the number of rats crossing the platform in the model group was significantly reduced （P＜0.05）， and the number of crossings of the platform in the electroacupuncture group increased significantly （P＜0.05）. The results of TTC staining showed that the volume of cerebral infarction increased clearly in the model group compared with the sham surgery group （P＜0.05）， and apparently decreased in the electroacupuncture group compared with the model group （P＜0.05）. Golgi staining showed that the number of dendritic branches of pyramidal neurons and dendritic spines in hippocampal CA1 region significantly decreased in the model group compared with the normal group and the sham surgery group （P＜0.05）. The number of dendritic branches of pyramidal neurons and the density of dendritic spines in hippocampal CA1 region significantly increased in the electroacupuncture group compared with the model group （P＜0.05）. The protein relative expression levels of GluR1， GluR2， GluR3， TARPγ2 and TARPγ8， and the mRNA levels of GluR1， GluR2 and GluR3 in hippocampus decreased in the model group compared with the normal group and the sham surgery group （P＜0.05）. The protein relative expression levels of GluR1， GluR2， GluR3， TARPγ2 and TARPγ8， and the mRNA levels of GluR1， GluR2 and GluR3 in hippocampus increased in the electroacupuncture group compared with model group （P＜0.05）. ConclusionThe Tongdu Xingshen needling method can improve learning memory impairment after cerebral ischaemia-reperfusion， which may be related to up-regulation of the expression of AMPA receptor and their auxiliary protein TARP， and promoting the synaptic plasticity of hippocampal tissues.
RESUMO
Objective:To investigate the expression of transmembrane 4 super family 1 (TM4SF1)in breast cancer tissue,and to elucidate its clinical significance and explore the related molecular biological mechanisms. Methods:A total of 190 cases of human breast cancer,110 cases of paracancerous tissue and 110 cases of normal breast tissue were collected.Immunohistochemistry was used to detect the expression levels of TM4SF1 mRNA in breast cancer tissue,paracancerous tissue,and normal breast tissue;Western blotting method was used to detect the expression levels of TM4SF1 in breast cancer tissue,paracancerous tissue,and normal breast tissue;RT-PCR method was used to detect the expression levels of TM4SF1 mRNA in breast cancer tissue,paracancerous tissue, and normal breast tissue.The positive expression rates of TM4SF1 in breast cancer tissue of the breast cancer patients with different clinicopathological features were detected.Results:The positive expression rate of TM4SF1 in the breast cancer tissue was significantly higher than those in paracancerous tissue and normal breast tissue (P <0.05);there was no significant difference in the positive expression rates of TM4SF1 between paracancerous tissue and normal breast tissue (P = 0.531);the expression of TM4SF1 was not correlated with age,but was closely correlated with tumor size,differentiation degree,lymph node metastasis and tumor stage (P <0.05);the positive expression rate of TM4SF1 in basal like breast cancer tissue was higher than those in the other three types of tissues (P <0.05).The results of Western blotting showed that the expression level of TM4SF1 in breast cancer tissue was higher than those in paracancerous tissue and normal breast tissue (P < 0.05 ), but there was no significant difference in the expression level of TM4SF1 between paracancerous tissue and normal breast tissue (P >0.05). The results of RT-PCR showed that the expression level of TM4SF1 mRNA in breast cancer tissue was higher than those in the paracancerous tissue and normal breast tissue (P <0.01);there was no significant difference in the expression level of TM4SF1 mRNA between paracancerous tissue and normal breast tissue (P > 0.05 ). Conclusion:The TM4SF1 is highly expressed in breast cancer tissue. TM4SF1 may affect the occurrence, development and distant metastasis of breast cancer through various mechanisms.TM4SF1 may be a potential target for the treatment of breast cancer.