AIM: To observe the effect of cannabinoid receptor (CB1R) agonist WIN55-212-2 on the expression of brain-derived neurotrophic factor (BDNF), c-Fos and protein kinase A beta-catalytic subunit (PKAC-β) in cerebrum cortex after intracerebral hemorrhage (ICH) in rats. METHODS: The intracerebral hemorrhage model of rat was made by the injection of collagenase Ⅶ, and WIN55-212-2 was intraperitoneally (ip) injected 30 min later. The rats were killed for sampling the brain tissues as specimens 24 h after ICH. The methods of immunohistochemical analysis and Western blotting were adopted to detect the expression of PKAC-β and BDNF. The mRNA expression of PKAC-β, c-Fos and BDNF was determined by RT-PCR. RESULTS: WIN55-212-2 obviously improved some nervous deficit symptoms and increased the expression of BDNF at mRNA and protein levels with upregulating the mRNA expression of c-Fos and downregulating the expression of PKA at mRNA and protein levels in the ipsilateral cerebral cortex. The proteins of PKAC-β, c-Fos and BDNF were expressed on the membrane or nucleus of the neuron or in the cytoplasm of glial cells, respectively. CONCLUSION: The expression of BDNF is induced not only by upregulation of c-Fos, but also by downregulation of PKA in WIN55-212-2 treated rats.

Recently,the progress in employing transplanting stem cells to cure injured retina is very fast and has been continuously yielding exciting results.Various sources are used in the studies,including retina-derived cells such as M?ller cells and ciliary body cells,and non-retina-derived cells such as embryonic stem cells and brain-derived stem cells.This review briefly discusses the recent progress of these studies.

Aim To explore the effects of cannabinoid (WIN55,212-2) on mRNA and protein expressions of PKAC-?,c-fos and BDNF in cerebral cortex after intracerebral hemorrhage(ICH)in rats.Methods ICH model of rats were made by Ⅶ Collagenase,which were injected by the intraperitoneal route,thirty minutes after the operations.The rats were killed for brain tissue as specimens with 24 hours.The localizations of PKAC-?,c-fos and BDNF were assessed by immunohistochemistry;The expressions of PKAC-?,c-fos and BDNF mRNA were detected by RT-PCR;The expressions of PKAC-? and BDNF protein were revealed by Western blot.Results WIN55,212-2 increased not only the levels of BDNF mRNA and protein,but also c-fos mRNA in ipsilateral cerebral cortex.However,it decreased the levels of PKAC-? mRNA and protein.PKAC-?,c-fos,and BDNF proteins were expressed on membrane of neurons,nucleus of neurons or the cytoplasm of glial cells respectively.Conclusion WIN55,212-2 induces the expression of BDNF in the cerebral cortex,which provides a theoretical basis for the treatment of cerebrovascular disease.

Aim To explore the effects of cannabinoid (WIN55,212-2) on mRNA and protein expressions of PKAC-β,c-fos and BDNF in cerebral cortex after intracerebral hemorrhage(ICH)in rats.Methods ICH model of rats were made by Ⅶ Collagenase,which were injected by the intraperitoneal route,thirty minutes after the operations.The rats were killed for brain tissue as specimens with 24 hours.The localizations of PKAC-β,c-fos and BDNF were assessed by immunohistochemistry;The expressions of PKAC-β,c-fos and BDNF mRNA were detected by RT-PCR;The expressions of PKAC-β and BDNF protein were revealed by Western blot.Results WIN55,212-2 increased not only the levels of BDNF mRNA and protein,but also c-fos mRNA in ipsilateral cerebral cortex.However,it decreased the levels of PKAC-β mRNA and protein.PKAC-β,c-fos,and BDNF proteins were expressed on membrane of neurons,nucleus of neurons or the cytoplasm of glial cells respectively.Conclusion WIN55,212-2 induces the expression of BDNF in the cerebral cortex,which provides a theoretical basis for the treatment of cerebrovascular disease.

Aim To study the changes of the biological behavior of dermal fibroblasts in Ⅲ skin burns wound in rats using chitosan.Methods Wistar rats were randomly divided into 5 groups as follows:1% chitosan(W/V)group,2% chitosan(W/V)group,4% chitosan(W/V)group,bFGF(basic fibroblast growth factor) group and the control group.Rats were made for Ⅲ skin burns.The wound healing time was recorded,and the wound healing rate was calculated.Then the cell cycle and apoptotic dermal fibroblasts were determined and the amount of Hydroxyproline(HOP) in the skin tissue was analyzed.Results The wound healing rate of 4% chitosan(W/V) group was higher and the wound healing time of 4% chitosan(W/V) group was shorter than that of the control group.On the 7th,14th day post-injury,the content of protein of 4% chitosan(W/V) group was higher than that of the control group.The content of HOP of 2% and 4% chitosan(W/V) group was highest on the 7th day post-injury. Compared with that in control group,the percentage of cells of S stage in 4% chitosan(W/V) group was aboundant,and was reduced in apoptotic dermal fibroblasts.Conclusion The changes of cellular biological behaviors might be one of the mechanisms of that Chitosan could promote the wound healing of Ⅲ skin burns in rats significantly.

0.05).It could also decrease the times of wet manure induced by folia sennae,while it was of no effect on diarrhea induced by castor oil. CONCLUSION: Volatile oil from A.longiligulare T.L.Wu has anti-inflammation,analgesic and anti-diarrhea effect related to the cure for ulcerative colitis.

AIM: To investigate antioxidative and antinitrosative effects of volatile oil from A.longiligulare T.L.Wu on dextran sulfate sodium(DSS)-induced ulcerative colitis mice. METHODS: Balb/c mice were fed with 4% DSS solution for 7 d to induce ulcerative colitis.Using biochemical method,the activity of antioxidative enzyme SOD,MDA and NO were determined in normal,model,SASP and three mouse's groups with low,moderate and high volatile oil from A.longiligulare T.L.Wu respectively.At the same time,the activity of iNOS was also measured by immunohistopathology. RESULTS: The concentration of MDA and NO were reduced and SOD increased significantly in high-and moderate-volatile oil groups compared with those in the model group.The activity of iNOS was reduced significantly in high-and moderate-volatile oil groups compared with those in the model group.The results demonstrated that the expression of iNOS was significantly inhibited in DSSinduced ulcerative colitis mice after being treated with high or moderate-dosage volatile oil. CONCLUSION: Volatile oil from A.longiligulare T.L.Wu has antioxidative and antinitrosative effects which may be one of the mechanism for treating UC.

BackgroundHypoxia and hyperglycemia are the common causes of retinal neovascularization.In these states,H+ accumulates because of the elevated glycolysis and failure of retinal circulation,thus the retinas readily acidified. ObjectiveThe present study was to explore whether retinal acidosis independently regulates the production of vascular endothelial growth factor (VEGF) and pigment epithelium-derived factor (PEDF) and whether the regulation is related to oxidative stress.Methods The retinas from 2-week-old male SD rats were cultured with explant method in DMEM modulated by NaHCO3,and culture retinas were randomly divided into pH 7.2,6.8 and 6.5 groups for 24 hours.In addition,after 24 hours of culture as above described,retinas were washed using PBS two times and then followed by again culture in DMEM with pH 7.2 for another 24 hours.Also,antioxidant was added in different pH values of DMEM for culture as above described.The retinal samples were prepared for histopathological examination.The expressions of VEGF and PEDF proteins and their mRNA in retina tissue were detected by Western blot and fluorescence quantitative polymerase chain reaction (PCR) respectively.Results The retina showed the clear structure and morphology in pH 7.2 group and pH 6.8 group,but retinal vacuoles change was seen in pH 6.5 group after culture for 24 hours.No significant difference was seen in the expressing level of VEGF mRNA in retina between normal group and pH 7.2 group( 112％ ±11％ vs 100％ ±7％ ) (P=0.55),but those in pH 6.8 group and pH 6.5 group were significant increased in comparison with pH 7.2 group( 196％ ±43％ vs 100％ ±7％ ;251％ ±29％vs 100％ ±7％ )( P＜0.05 ).The expressing level of PEDF mRNA in retina in normal group was similar to that of pH7.2 group(86％ ±19％ vs 100％ ±33％) (P=0.64),but that in pH 6.5 group was significantly higher than pH 7.2 group( 230％ ±66％ vs 100％ ±33％ ) ( P＜0.05 ).The resemble results were found in the expressions of VEGF and PEDF protein.After pH reversion,the expressing levels of VEGF mRNA were 100％ ±13％,111％ ±9％,113％ ±9％ in pH 7.2 group,pH 6.8 group and pH 6.5 group respectively without significant difference among them (F=2.51,P=0.16).The expressing levels of PEDF mRNA were 100％ ±13％,110％ ±9％,108％ ±11％in different groups ( F =0.98,P =0.43 ).Under the presence of antioxidant,the expressing level of VEGF mRNA in pH 6.5 group increased in comparison with pH 7.2 group and pH 6.8 group ( P ＜ 0.05 ).The expressing levels of PEDF mRNA were significant different among pH 7.2 group( 100±31 )％,pH 6.8 group( 282±45 )％ and pH 6.5 group(480±117)％ (F=20.73,P=0.00). Conclusions VEGF can be induced by retinal acidification alone,which may be regulated by oxidative stress.Under the retinal acidification,antioxidants promote the expression of PEDF,suggesting that oxidative stress inhibits the production of PEDF.