The rapid screening of bioactive constituents within traditional Chinese medicine (TCM) presents a significant challenge to researchers. Prevailing strategies for the screening of active components in TCM often overlook trace components owing to their concealment by more abundant constituents. To address this limitation, a fishing strategy based on offline two-dimensional liquid chromatography (2D-LC) combined with surface plasmon resonance (SPR) was utilized to screen bioactive trace components targeting peroxiredoxin 3 (PRDX3), using Uncaria alkaloids (UAs) as a case study. Initially, an orthogonal preparative offline 2D-LC system combining a positively charged C₁₈ column and a conventional C₁₈ column under disparate mobile phase conditions was constructed. To fully reveal the trace alkaloids, 13 2D fractions of UAs were prepared, and their components were characterized using mass spectrometry (MS). Subsequently, employing PRDX3 as the targeting protein, a SPR-based screening approach was established and rigorously validated with geissoschizine methyl ether (GSM) serving as a positive control for binding. Employing this refined strategy, 29 candidate binding alkaloids were fished from the 13 2D fractions. Notably, combining offline 2D-LC with SPR increased the yield of candidate binding components from 10 to 29 when compared to SPR-based screening alone. Subsequent binding affinity assays confirmed that PRDX3 was a direct binding target for the 12 fished alkaloids, with isovallesiachotamine (IV), corynoxeine N-oxide (CO-N), and cadambine (CAD) demonstrating the highest affinity for PRDX3. Their interactions were further validated through molecular docking analysis. Subsequent intracellular H₂O₂ measurement assays and transfection experiments confirmed that these three trace alkaloids enhanced PRDX3-mediated H₂O₂ clearance. In conclusion, this study introduced an innovative strategy for the identification of active trace components in TCM. This approach holds promise for accelerating research on medicinal components within this field.

Objective To explore the significance of Survivin and PTEN in gastric cancer development and correlation between Survivin and PTEN.Methods Immunohistochemical methods were applied to detect the expres.sion of Survivin and PTEN in 105 pathological specimens of gastric cancer, in comparison to that in 98 negative gastric mucosa tissues.Results The positive rate of Survivin expression was 82.9% (87/105) in tissue specimens of gastric cancer and 6.1% (6/98) in tissue specimens of non-gastric cancer;The difference between them was statistically sig nificant (P <0.01).The positive rates of PTEN expression were 34.3% (36/105) in tissue specimens of gastric canc-er and 99.0% (97/98) in tissue specimens of non-gastric cancer;The difference between them was statistically signifi-cant ( P <0.01).The expression of Survivin and PTEN in tissues of gastric cancer had nothing to do with the gender of patients ( P > 0.05 ) , but was correlated with the degree of tumor cell differentiation, depth of invasion, lymph node me-tastasis, distant organ metastasis and TNM staging(P <0.05).Analysis by Spearman showed that Survivin was nega-tively correlated with PTEN( r = -0.23 ,P < 0.05 ).Conclusion The incidence of gastric cancer was related to the abnormally high level of Survivin expression and absence of PTEN expression;The expression of Survivin and PTEN in tissues of gastric cancer was unrelated with the gender of patients, but related to the degree of tumor cell differentia-tion, depth of invasion,lymph node metastasis,distant organ metastasis and TNM staging.The expression of Survivin was negatively correlated with the expression of PTEN.

0.05). The cutaneous part occupied 2/3 of the entire upper lip height and the vermilion 1/3.The cutaneous part occupied more than 1/3 of the entire lower lip height.The chin occupied more than 60 percent of the entire mandibular height. Conclusion: The results of the study are consistent with the date of Farkas.