Objective To study the optimum conditions for extracting total flavonoids from Acanthus illicifolius through the two extraction processes.Methods An orthogonal test of L_9 (3~4)was designed to select optimum operation conditions for extracting total flavonoids from Acanthus illicifolius through Soxhelt and ultrasonic extraction process.According to the output of total flavonoids under the different extraction conditions,which were selected as ethanol contents,extraction temperatures and extraction time for Soxhelt extraction process, and extraction time,ethanol contents and ratio of liquid to material for ultrasonic extraction process.Results The optimum conditions were established as follows:ethanol content 60%, extraction temperature 85℃and extraction time 2.5h in Soxhlet extraction process;Ethanol content 50%,extraction time 50min,ratio of liquid to material 40:1 in ultrasonic extraction process.The extraction temperature was the main factor on the Soxhlet extraction process, and the ethanol content on the ultrasonic extraction process.Conclusion The technology of ultrasonic extraction is better than that of Soxhlet extraction to extract the total flavonoids from Acanthus illicifolius.The max output of total flavonoid from Acanthus illicifolius can reach to 3.82%.

Objective:To study the utility of immunohistochemistry (IHC) in differential diagnosis between trichoblastoma (TB) and basal cell carcinoma (BCC).Methods:Fifty-eight cases of TB and 40 cases of BCC were collected at Fudan University Shanghai Cancer Center from January 2009 to December 2019 and retrospectively analyzed by IHC for bcl-2, Ber-EP4, CD10, CK20 and Ki-67. Fisher exact test was performed for statistical analysis.Results:Twenty-five (43.1%) TBs and 5 (12.5%) BCCs showed bcl-2 staining in the outermost layer of the epithelial nests, the difference was statistically significant ( P<0.01). The proportion of cases with bcl-2 staining>75% of epithelial cells in BCC group was much higher than that in TB group (40% vs. 12.1%; P<0.01). BCC group showed larger proportions with Ber-EP4 staining>75%, 51%-75% of epithelial cells than TB group (12.5% vs. 1.7%, 37.5% vs. 8.6%; P<0.05). Fifty-five (94.8%) TBs demonstrated CD10 expression in the follicular stroma, while only 16 (40.0%) BCCs showed focal or scattered CD10 expression in reactive fibrous stroma ( P<0.01). CK20 expression was present in 37 (63.8%) TBs with scattered pattern, but BCCs exhibited no CK20 staining except for only one case (2.5%) showing focal staining ( P<0.01). Compared with TB group, the BCC group included more cases with Ki-67 labeling index ≥15% on average and ≥25% in hotspot areas ( P<0.05). Conclusion:IHC is helpful in differential diagnosis between TB and BCC. Scattered CK20 staining pattern and stromal CD10 expression support the diagnosis of TB. Bcl-2 staining limited to the outermost layer of the proliferation is more likely to be found in TB. In contrast, Ber-EP4 positivity and higher Ki-67 labeling index tend to be present in BCC.