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Young leaves of Kabuli chickpea as well as soybean Xiangdou No.3, which are the current plants that studied in our laboratory were selected as materials. Effects on protoplasts yield and survival rate of different enzyme combination, concentration of D-Mannitol in enzyme combinations, pH of enzyme combinations and enzymolysis time are detected. The results showed that, the best condition for Xiangdou No.3 leaf protoplasts isolation is to rotate the cut materials for 6 hours in enyzme solution under temperature of 27 ℃ and rotate speed of 45 r/min for 6 h. Onozuka R-10 (0.5%), Hemicellulase (0.8%), Macerozyme R-10 (0.8%) in combination with Pectolyase Y-23 (0.4%) dissolving in CPW solution with MES (0.1%) and Mannitol (10%), pH 6.0 was found best for protoplasts isolation of Xiangdou No.3 leaves.The best condition for protoplasts isolation of Kabuli chickpea is to put the cut materials into enzymatic hydrolysate enzymolyse for 7 to 8 hours under temperature of 27 ℃ and rotate speed of 45 r/min on water bath shaker, the optimum combination of enzyme consists of Onozuka R-10 (0.5%), Hemicellulase (0.8%), Macerozyme R-10 (0.8%), MES (0.1%) and Mannitol (10%) dissolved in CPW solution with pH 4.8. The protoplasts prepared with the methods above are used in subcellular location and the effects show well.
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Objective To investigate the effect of acupuncture on gap junction protein Cx43 expression in the hippocampal region in ischemia/reperfusion rats.Methods Wistar rats were randomized into normal, model, non-point acupuncture and acupuncture groups. A rat model of focal cerebral ischemia/reperfusion was made by modified middle cerebral artery thread occlusion. The model, non-point acupuncture and acupuncture groups were separately divide into four subgroups: 30, 60, 180 and 360 min ischemia/reperfusion, 10 rats each. The middle and lateral lines of vertex were given electroacupuncture in the acupuncture group of rats. A subcostal fixed point 10 mm above the iliac crest on the affect side was selected as an acupuncture point in the non-point acupuncture group. The model group was not treated. Cx43 protein expression was determined by an immunohistochemical method.Results There was a statistically significant difference in the behavior disorder (Bederson’s) score between the acupuncture group and the non-point acupuncture or model group after different times of cerebral ischemia/ reperfusion (P<0.05). There was a statistically significant difference in hippocampal Cx43 content between model, acupuncture or non-point acupuncture group and the normal group and between the acupuncture group and the model or non-point acupuncture group after different times of cerebral ischemia/reperfusion (P<0.05).Conclusions Acupuncture can inhibit the overexpression of Cx43, intervene in cerebral ischemia-reperfusion injury and produce a neuroprotective effect in ischemic brain injury.
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The standardization of acupuncture and moxibustion in Canada is increasingly flouring nowadays:on one hand,acupuncture industry in legislated provinces is in the charge of direct administrative departments and associations of acu-moxi; on the other hand,according to the local situations,relevant regulations have been made,which are concentrated on two aspects-technical and administrative standards.Undoubtedly,Canadian Alliance of Regulatory Bodies for TCM Practitioners and Acupuncturist (CARB),united by each provincial TCM/Acupuncture Administration,is the first step to issue federal standards and establish federal level management institution.However,now the releasing of a fundamental regulatory file is in need of active corporation and publicity from each province and association.Therefore,it shows that the body of standardization in Canada is still not relatively perfect.Through introducing and analyzing the relevant information,the author expects that the readers can know the current development of acupuncture and moxibustion standardization in Canada.
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The development of acupuncture in Australia has been booming.Under this circumstance,a series of acupuncture standards,codes and guidelines were established by different associations of TCM in Australia.Moreover,Australia has set the front pace in the field of acupuncture standards among the countries outside China.This paper is to introduce the acupuncture standards of Australia in three aspects respectively:national standards,industry standards and regional standards and to generalize the content of the main standards as well as analyze the current condition of Australian acupuncture standardization in order to generally give an all-around view of Australian acupuncture standards to the researchers in the field of international acupuncture standards.
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ObjectiveTo observe the effects of Huatan Tongluo on the Bederson's scores,the permeability of blood-brain barrier and ultrastructure of hippocampus endothelial cells after thrombolysis in acute cerebral infarction.To provide experimental evidence for the prevention of bleeding after thrombolysis in acute cerebral infarction with Huatan Tongluo.Methods 120 SD male rats were randomly divided into sham group,model group,rt-PA group,Huatan Tongluo combined rt-PA low-dose group,Huatan Tougluo combined rt-PA medium-dose group and Huatan Tongluo combined rt-PA high-dose group( each group was divided in 6h,24h,3d,7d four-phase).MCAO rats were prepared by self-embolus method.rt-PA group and Huatan Tongluo combined rt-PA groups were given rt-PA through tail vein injection and three doses of Huatan Tongluo to treat.To observe the Bederson's scores in rat on four-phase.To detect the permeability of blood-brain barrier(BBB) by the Evans Blue dye ( EB dye) and watch the ultrastructure of hippocampus endothelial cells in ischemic region by electron microscopy.Results 1.Bederson's scores:Compared with model group,rt-PA group,Huatan Tongluo combined rt-PA low-dose group,Huatan Tongluo combined rt-PA medium-dose group and Huatan Tongluo combined rt-PA high-dose group significantly lower(P＜0.01 ) ; Compared with rt-PA group,Huatan Tongluo combined rt-PA medium-dose groups and high-dose groups was significantly lower(P ＜ 0.05).2.EB dye:Compared with model group,rt-PA group,Huatan Tongluo combined rt-PA low-dose group,Huatan Tongluo combined rt-PA medium-dose group and Huatan Tongluo combined rt-PA high-dose group (P ＜ 0.01 ) ; Compared with rt-PA group ( 6h:( 236.72 ± 79.79 ) ng/g; 24h:(229.72 ±22.18)ng/g;3d:(163.34 ±24.45)ng/g;7d:(133.21 ±25.09)ng/g),Huatan Tongluo combined rt-PA medium-dose groups (6h:( 121.63 ± 33.27) ng/g;24h:( 153.78 ± 60.67 ) ng/g;3d:( 76.05 ± 27.11 ) ng/g;7d:(72.97 ±25.81 ) ng/g) and high-dose groups (6h:( 123.11 ±23.65) ng/g;24h:( 138.36 ±52.36)ng/g;3d:(83.00 ±32.00)ng/g;7d:(75.06 ± 13.97) ng/g) was significantly lower(P＜ 0.01 ).3.Electron microscopy:Compared with model group,treatment groups all protected the endothelial cells.In 24h,the Huatan Tongluo combined rt-PA high-dose group was best.At 3d and 7d,the Huatan Tongluo combined rt-PA medium-dose group was best.ConclusionHuatan Tongluo combined rt-PA to thrombolysis can protect the blood-brain barrier and endothelial cells of hippocampal's ischemic region.After thrombolytic,to adjust the medicine drug concentration at different time can play a better treatment results.
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Objective:To investigate the pathological changes following the focal cerebral ischemia and reperfusion in rats and the effect of acupuncture on them. Methods: A model of ischemia and reperfusion was established by thread occlusion, and the brain-activating acupuncture was applied, and then the morphological changes of the cerebral cortex on the ischemic side were observed under electron and light microscopes. Results:Cerebral ischemia and reperfusion could lead to the structural injury of neurons, gliacytes and capillary vessels in rat's brain, and acupuncture could improve the ultra-structural injury in the area around the cerebral ischemia. It was also found that acupuncture intervention at 3 hours following reperfusion was more ideal than that at any other time points. Conclusion: Acupuncture has a protective effect on the ultra-structural injury of neurons associated with focal cerebral ischemia and reperfusion in rats, and acupuncture given within 3 hours following reperfusion can produce a satisfactory effect.