Acetylcholinesterase is an important target for anti-Alzheimer's disease now.Much attention has been paid to the development of tacrine dimeric or hybrid compounds.The design strategy is transforming merely interacting with the two binding sites of acetylcholinesterase to get more activity and selectivity into the multi-target-directed function for the treatment of Alzheimer's disease. Herein,the tacrine dimeric or hybrid compounds development and the prospect for the treatment of AD have been reviewed.

AIM: To confirm the differential expression genes in the rat ischemic brain. METHODS: The middle cerebral artery occlusion ischemic model was set up in rats. Fluorescence differential display reverse transcriptase-polymerase chain reaction (FDD RT-PCR) and reverse Northern blotting were used to fast confirm the differential expression genes. RESULTS: Nine differential expression sequence tags, including 6 known sequences and 3 unknown sequences, were confirmed. Among the known sequences, mus musculus ab1-interactor1,homo sapiens CGI-99 protein, tissue inhibitor of metalloproteinase 3 and homosapiens nuclear receptor co-repressor were up-regulated while homo-sapiens nuclear matrix protein p84 and coatomer protein complex, subunit gamma 2 were down-regulated. CONCLUSIONS: ① Combination of fluorescence differential display reverse transcriptase-polymerase chain reaction (FDD RT-PCR) and reverse Northern blotting is a method to fast-confirm the differential expression genes; ② There are differential expression genes in ischemic brain regions compared to non-ischemic parts. [

AIM: To Screen and identify differentially expressed genes that involved in apoptosis model in rat cerebellar granule neurons (CGNs).METHODS: The rat cerebellar granule neurons were isolated and primarily cultured. Fluorescent differential display RT-PCR (FDD RT-PCR) was performed to screen differentially expressed ESTs in the apoptosis model of primarily cultured rat CGNs. ESTs were subcloned into pGEM-T EasyTM vector and then sequenced. Alignment assay in non-redunant database was applied for encoding information. Reverse Northern blotting was used to appraise the results from DDRT-PCR.RESULTS: 164 pieces of differentially expressed ESTs were obtained by FDDRT-PCR. 17 of them were subcloned and sequenced. 5 ESTs of 17 were confirmed to be positive results by reverse Northern blotting. CONCLUSION: DD-PCR is a rapid, simple-operation and sensitive method for screening differentially expressed genes, which would contribute to the molecular mechanisms of apoptosis/survive of CGNs.

Objective To investigate the regulations of Bax , Bcl-2 in the protection of lipoic acid-niacin diad in acrolein-induced apoptosis in ARPE-19 cells. Methods The ARPE-19 cells were cultured in medium containing 10% fetal bovine serum , at 37 ℃ with 5% CO2. The ARPE-19 was transferred to 6-well plate after reaching to 70% confluence. After starvation for 24 h , the cells in 6-well plates were divided into three groups , including the blank control group , the acrolein treatment group with 50 μmol/L acrolein for 24 h , and the protection group with 100 μmol/L lipoic acid-niacin diad for 24 h and with the acrolein for another 24 h. The apoptotic cells were detected by flow cytometry assay , and expressions of Bcl-2 , Bax protein were detected by Western Blot assay. Results The percentages of normal healthy cells were 94.8%, 60.98%, and 91.34% in the blank control group , 50 μmol/L acrolein group and 100 μmol/L diad contained of lipoic acid and niacin group , respectively. The ratios of Bax/Bcl-2 protein expression were 0.293 9, 1.389 2, and 0.555 8 in the blank control group, 50 μmol/L acrolein group and 100 μmol/L diad contained of lipoic acid and niacin group, respectively. Conclusion The protective effect of lipoic acid-niacin diad on acrolein-induced apoptosis in ARPE-19 cell through promoting Bcl-2 expression and inhibiting Bax expression.

Objective To explore the changes of cell proliferation and differentiation of neural stem cells induced by fasudil treatment,and to primarily study the mechanism in C17.2 mice.Methods C17.2 cells were cultured in vitro; 5,25,50 and 100 μmol/L fasudil were given to the cells,respectively,for 24 h,and cells in the blank control group were given the same volume of culture medium.The changes of cell morphology were observed under a phase-contrast microscope; cell viability and cell necrosis rate were determined by MTT assay and lactate dehydrogenase (LDH) assay,respectively.Western blotting was applied to detect the expression levels of neural markers (nestin,glial fibrillary acidic protein [GFAP],double cortisol [DCX],microtubule-associated protein-2 [MAP-2]),and Notch Ⅰ and Hes 1 proteins in the notch signaling in cells from the 100 μmol/L fasudil treatment group and blank control group.Immunofluorescence staining was used to detect the nestin and GFAP expressions in the C 17.2 cells.Results As compared with that in the blank control group,the cell viability in the 50 and 100 μmol/L fasudil treatment groups was significantly decreased; that in the 100 μmol/L fasudil treatment group was significantly lower than that in 50 μmol/L fasudil treatment group (P＜0.05); LDH assay showed no significant difference of cell necrosis among the five groups (P＜0.05).Western blotting indicated that 100 μmol/L fasudil treatment group had significantly decreased nestin expression,significantly elevated DCX,MAP-2 and GFAP expressions,and statistically decreased expression levels of Notch 1 and Hes 1 as compared with blank control group (P＜0.05).Immunofluorescence staining indicated that the percentage of nestin positive cells was markedly decreased,the percentage of GFAP positive cells was significantly increased in the 100 μmol/L fasudil treatment group as compared with those in the blank control group (P＜0.05).Conclusion Fasudil treatment could inhibit the proliferation of C17.2 cells and promote them differentiate into neuronal and glial cells via decreasing the expression level of Notch signaling.