Objective to investigate the effect of berberine on the expression of AQP4 and neuronal injury after focal cerebral ischemia in diabetic rats. Methods Male SD rats were randomly divided into four groups:control(sham surgery),middle cerebral artery occlusion and reperfusion (MCAO/R),MCAO/R treated with vehicle(DMSO),MCAO/R treated with berberine. the transient focal ischemia/reperfusion was induced by in-troducing silicone-coated monofilament nylon suture from the right external carotid artery into the origin of the middle cerebral artery,which was re-moved after 60 min. In group treated with berberine,the rats were injected with berberine before and after suffered from cerebral ischemia. Similarly, in group vehicle,the animals received DMSO vehicle at the same time. the score of neurological behavior was evaluated 24 h after reperfusion. Mean-while,the rats were sacrificed for Nissl staining. to estimate cerebral edema,the wet-dry ratio was measured. the expression of AQP4 in the border of the infarct region in different groups was observed by Western blot. Results Compared with the model group,berberine improved neurological deficits(P < 0.05). Berberine treatment inhibited the neuronal deformation shown by Nissl staining(P < 0.05). Berberine significantly decreased the wet-dry ratio and reduced the expression of AQP4(P < 0.05). Conclusion these results suggested that berberine could induce neuroprotection against ischemic injury by inhibiting the expression of AQP4 in diabetic rats.

Objective To investigate the effects of nerve growth factor (NGF) on retinal synaptic plasticity of diabetic mellitus rat and its underlying mechanisms.Methods A total of 24 clean SD male rats were randomly divided into three groups (n =8),and they were control group,diabetic group and treatment group.In the latter two groups,a model of diabetic rats was induced by streptozotocin,and then the rats of treatment group were injected intraperitoneally 800 U · kg-1 NGF once a day after the model was induced successfully.Both control group and diabetic group were given the same amount of normal saline.Twelve weeks later,MDA content and SOD activity were detected;meanwhile,the expression of retinal synaptophysin was detected by immunofluorescence,and the expressions of retina synaptophysin and Caspase-3 were detected by Western blot.Results The difference of MDA content in the three groups was statistically significant (F =85.46,P ＜ 0.01);and the content of MDA in the diabetic group was significantly higher than that in the control group (P ＜0.01),while its content in the treatment group was significantly lower than that in the diabetic group (P ＜0.01).The difference of SOD activity in the three groups was statistically significant (F =17.76,P ＜0.01);and the SOD activity in the diabetic group was significantly lower than that in the control group (P ＜0.01),while its activity in the treatment group was significantly higher than that in the diabetic group (P ＜0.01).The difference of immunofluorescence intensity of synaptophysin in the three groups was statistically significant (F =395.42,P ＜ 0.01);immunofluorescence intensity of synaptophysin in the diabetic group was attenuated compared with the control group (P ＜0.01),while the intensity in the treatment group was enhanced compared with the diabetic group(P ＜0.01).The difference of the relative expression of synaptophysin in the three groups was statistically significant (F =17.27,P ＜ 0.01);and the expression of synaptophysin in the diabetic group was significantly downregulated compared with the control group (P ＜ 0.01),while its expression in the treatment group was upregulated compared with the diabetic group (P ＜ 0.01).The difference of relative expression of Caspase 3 protein in the three groups was statistically significant (F=217.13,P ＜0.01);and the expression level of Caspase 3 in the diabetic group was significantly higher than that in the control group (P ＜0.01),while its level in the treatment group was significantly lower than that in the diabetic group (P ＜ 0.01).Conclusion NGF can help to inhibit the apoptosis of retinal cell,restore the number of retina synapse by reducing the oxidative stress in diabetic retina,which suggests that NGF may be involved in the changes of synaptic plasticity in diabetic retina via oxidative stress pathway.

Objective To optimize the volatile oil extraction and inclusion process of Wenweiyang capsules. Methods An orthogonal test was adopted in this study. The extraction technology was optimized for the yield of volatile oil regarding the amount of water loaded, grain size of medicinal material, and decoction time as factors. The inclusion technology was optimized for the inclusion yield and volatile oil inclusion rate using the ratio ofβ-CD:oil, amount of water and grinding time as factors. Results The optimized extraction parameters were as follows:breaking medicinal material through 10 mesh screen, adding 6 fold volume of water and extracting for 5 h. The optimized inclusion progress was grinding at theβ-CD:oil ratio of 81, loading equivalent amount of water and grinding for 30 minutes. The average yield of volatile oil is 1. 72%, the average inclusion rate is 93. 01% and the average volatile oil inclusion rate is 74. 82%. Conclusion The extraction and inclusion technology is simple, reliable, which can effectively retain the volatile oil and provide evidence for the preparation of Wenweiyang capsules.

Objective To investigate the effects of inhibiting NgR on retinal ganglion cells density and synaptophysin expression of diabetic rat.Methods Thirty-two SD male rats were randomly divided into normal control group,diabetic group,siNgR group and scNgR group,8 rats in each group.Normal control group was given no any treatment.Diabetes model was induced by intraperitoneal injection of 50 mg · kg-1 streptozotocin in diabetic group,siNgR group and scNgR group,and the blood giucose more than 16.7 mmol · L-1 at 72 hours was set as the successfully model.The rats of siNgR group were intravitreally administrated with anti-NgR nucleotide and the rats of scNgR group intravitreally administrated with negative nucleotide.Eight weeks later,HE staining was conducted to detect density of retinal ganglion cell (RGC),immunofluorescence was used to evaluate the expression and distribution of synaptophysin (a marker of synaptic number).Relative expression of NgR and synaptophysin in retina were analyzed by Western blot.Results RGC density in normal control group,diabetes group,siNgR group and scNgR group were (624.33 ± 3.51) mm-2,(420.00 ± 2.65) mm-2,(621.67 ± 1.53) mm-2,(416.67 ± 2.52) mm-2,respectively.There was significant difference among four groups (F =5985.37,P ＜ 0.01).Compared with normal control group,RGC density in diabetes group and scNgR group were obviously decreased (all P ＜0.01),but siNgR group had no obviously change (P ＞ 0.05).The synaptophysin mainly expressed in the inner and outer network layer.Compared with normal control group,the positive expression of synaptophysin in diabetes group and scNgR group were decreased,but siNgR group had no obviously change.The relative expression of NgR in normal control group,diabetes group,siNgR group and scNgR group were (11.26 ±0.02) ％,(19.38 ± 0.10) ％,(11.17 ± 0.02) ％,(19.47 ± 0.31) ％,respectively.There was significant difference among four groups (F =2466.09,P ＜ 0.01).Compared with normal control group,the relative expression of NgR in diabetes group and scNgR group were obviously decreased (all P ＜ 0.01),but siNgR group had no obviously change (P ＞0.05).The relative expression of synaptophysin in normal control group,diabetes group,siNgR group and scNgR group were (35.76 ± 0.15) ％,(25.47 ± 0.36) ％,(35.28 ± 0.12) ％,(25.03 ± 0.75) ％,respectively.There was significant difference among four groups (F =583.70,P ＜ 0.01).Compared with the normal control group,the expression of synaptophysin in diabetic group and scNgR group were decreased increased (all P ＜ 0.01),while there was no significant difference in siNgR group (P ＞ 0.05).Conclusion Inhibiting the expression of NgR in the retina of diabetic rats can help to restore the number of synapses and protect the damaged RGC.

Objective To optimize the forming process of Wuzibushen Capsules. Methods Category and ratio of accessories were investigated by taking moisture absorption percentage as index. The inspected angle of repose, bulk density and critical relative humidity were also investigated. Results Starch was used as the excipien. Remedium cardinale and starch in the ratio of 1.15∶1.25 was more appropriate, 60% alcohol was added, dried at 60 ℃. Granules had a good fluidity, critical relative humidity was about 62%. Conclusion The forming technology was reasonable and provide reliable basis for production.