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Objective To investigate the effect of cluster needling at scalp points plus gavage of modified Buyang Huanwu decoction on NSC differentiation in rats with ischemic stroke of qi deficiency and blood stasis type.Methods A model of qi deficiency and blood stasis syndrome was made using healthy rats. Dil dye was given for pre-labelling after the success of model making. The rats were randomized into groups A, B, C, D, E and F. Group F consisted of 4 rats and each of the other groups, of 12 rats. A rat model of ischemic stroke of qi deficiency and blood stasis type was made by MCAO at 48 hrs after pre-labelling. After the success of model making, group A was the model group, group B was treated by cluster needling at scalp points, group C was treated by an oral gavage of Buyang Huanwu decoction, group D was treated by an oral gavage of modified Buyang Huanwu decoction, group E was treated by cluster needling at scalp points plus an oral gavage of modified Buyang Huanwu decoction and group F was the sham operation group. The rats were sacrificed at various time points respectively and the materials were taken. By nerve function assessment, double immunofluorescence staining and laser scanning confocal microscopy, a comparative study was conducted to investigate the effects of different treatments on NSC differentiation.Results There was a statistically significant difference in the nerve function score between group B, C, D, E or F and group A after one week of treatment (P<0.01), between group B or E and group A after two weeks of treatment (P<0.05,P<0.01) and between group E and group C or D after one and two weeks of treatment (P<0.01,P<0.05). After one and two weeks of treatment, there were statistically significant differences in Brdu﹢/NeuN﹢ cell count, Brdu﹢/GFAP﹢ cell count, Dil﹢/Brdu﹢NeuN﹢ cell count and Dil﹢/Brdu﹢/GFAP﹢ cell count between group B, C, D or E and group A (P<0.01), between group D or E and group B or C (P<0.01) and between groups E and D (P<0.01).Conclusions Various treatments have a promoting effect on nerve stem cell differentiation in the rats. Of various treatments, cluster needling at scalp points plus gavage of modified Buyang Huanwu decoction has the best therapeutic effect.
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OBJECTIVE@#To investigate the effect of inhibiting high mobility group box-1 (HMGB1) gene expression on the invasion and migration of endometrial carcinoma HEC-1A cells by small hairpin RNA.@*METHODS@#Three specific recombinant plasmids of HMGB1 (pshRNA-1 /HMGB1, pshRNA-2 / HMGB1, and pshRNA-3/HMGB1) were transfected into the endometrial cancer cell lines HEC- 1A by lipofectamine (TM) 2000. The expression of HMGB1 mRNA and protein was decteted by RTPCR and Western blot. The invasion and migration abilities of transfected HEC-1A cells were evaluated using Transwell assay and wound healing assay.@*RESULTS@#RT-PCR and Western blot revealed that the expression of HMGB1 at both mRNA and protein levels was significantly inhibited by HMGB1-pshRNA targeting sequence 1, 2, and 3 (P<0.05), and the levels of 3 mRNAs in the transfection group were 0.192±0.006, 0.055±0.002, and 0.123±0.086, respectively, which were significantly lower than those in the Lipo group (0.268±0.008) and the HMGB1/p-NC group (0.270±0.004). The maximum inhibiton rates of the 3 mRNAs were 28.4%, 79.5%, and 54.1%. The levels of 3 HMGB1 proteins in the transfection group were 0.259±0.129, 0.032±0.002, and 0.104±0.007, significantly lower than those in the Lipo group (0.347±0.007) and the HMGB1/p-NC group (0.349±0.007), and the maximum inhibitory rates were 25.4%, 90.8%, and 70.0%. The transwell chamber assay and wound healing assay showed that the invasion and migration of HEC-1A cells were effectively suppressed by inhibiting HMGB1 expression (P<0.05).@*CONCLUSION@#pshRNA-HMGB1 can effectively inhibit HMGB1 expression at both mRNA and protein levels, and decrease the invasion and migration of endometrial cancer cells.