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 establish a quality control method for the standard decoction of Polygoni Avicularis Herba.Methods:Totally 12 batches of decoction pieces from different origins were collected, the standard decoction was prepared and the quality evaluation method was established, the content of index components in the decoction pieces and the standard decoction was determined with HPLC, the index components, solution pH and other parameters were calculated, and the similarity analysis was carried out against the fingerprints.Results:The total content of myricetin in 12 batches of decoction pieces was >0.12%, and the content of myricetin in the standard decoction was >0.03%, which met the standard of the 2020 edition of the Chinese Pharmacopoeia. The pH value was 5.1-5.5, the transfer rate of myricetin components ranged from 50.0%-106.3%, and the fingerprint study showed that there were 7 common peaks. The similarity analysis results indicated that the standard decoction of 12 batches of decoction pieces of Polygoni Avicularis Herba had good consistency.Conclusion:The preparation process is stable and feasible in line with the traditional decoction preparation method, and can be used for the research and quality evaluation of the standard decoction.

Immunoglobulin (IgG) glycosylation affects the effector functions of IgG in a myriad of biological processes and has been closely associated with numerous autoimmune diseases, including systemic lupus erythematosus (SLE), thus underlining the pathogenic role of glycosylation aberration in autoimmunity. This study aims to explore the relationship between IgG sialylation patterns and lupus pregnancy. Relative to that in serum samples from the control cohort, IgG sialylation level was aberrantly downregulated in serum samples from the SLE cohort at four stages (from preconception to the third trimester of pregnancy) and was significantly associated with lupus activity and fetal loss during lupus pregnancy. The type I interferon signature of pregnant patients with SLE was negatively correlated with the level of IgG sialylation. The lack of sialylation dampened the ability of IgG to suppress the functions of plasmacytoid dendritic cells (pDCs). RNA-seq analysis further revealed that the expression of genes associated with the spleen tyrosine kinase (SYK) signaling pathway significantly differed between IgG- and deSia-IgG-treated pDCs. This finding was confirmed by the attenuation of the ability to phosphorylate SYK and BLNK in deSia-IgG. Finally, the coculture of pDCs isolated from pregnant patients with SLE with IgG/deSia-IgG demonstrated the sialylation-dependent anti-inflammatory function of IgG. Our findings suggested that IgG influences lupus activity through regulating pDCs function via the modulation of the SYK pathway in a sialic acid-dependent manner.
Humans ; Pregnancy ; Female ; Lupus Erythematosus, Systemic/pathology* ; Signal Transduction ; N-Acetylneuraminic Acid/metabolism* ; Immunoglobulin G ; Dendritic Cells/pathology*

Adult ; Atherosclerosis ; Cardiovascular Diseases ; China ; Habits ; Humans ; Lipids

Objective: To understand the longitudinal changes of refractive errors in adolescent myopia screening in the suburb of Shanghai, and to provide reference for targeted measures of myopia prevention and intervention. Methods: By using the cluster sampling method, 1 346 students were selected from two primary schools in a town in the suburb of Shanghai. Physical development indicators and refractive examination parameters of non ciliary muscle paralysis, and uncorrected visual acuity in 2017 and 2020 were collected from the Shanghai adolescent refractive development file. Longitudinal change of spherical equivalent (SE) refractive were assessed. A linear regression model was used to examine the relationship between the rate of SE change with characteristics of the students. Results: The average annual incidence of myopia was 16.36%, and the SE degrees of the left and right eyes of myopia students decreased by 225 degrees for three years. Girls (right eye Z=-4.33; left eye Z=-3.75, P<0.01), newly-onset myopia and persistent myopia (right eye Z=634.45; left eye Z=638.85, P<0.01) was a key for the rapid progress of refractive power. Conclusion The proportion of students with severe low vision is relatively high, and the apparent shifts toward more hyperopia in myopia students call for effective prevention and control programs based on changes in refractive to slow the progression of adolescent myopia.

Objective:To investigate the expression of mBmi-1 in papillary thyroid cancer (PTC) and to analyze its relationship with clinical features of PTC and its mechanism.Methons:A total of 60 tissue samples were collected from Jun. 2018 to Dec. 2018 in Department of Oncology Surgery of Affiliated Hangzhou First People’s Hospital, Zhejiang University School of Medicine. Real-time quantitative polymerase chain reaction (RT-PCR) was used to detect the expression level of B cell-specific MLV integration site-1 (mBmi-1) in PTC tissues and its relationship with pathological features was analyzed. Bmi-l microRNAs (miRNAs) were predicted by bioinformatics sites targetscan and microRNA.org and confirmed by luciferase reporter gene experiments. CCK-8 (Cell Counting Kit-8) kit and Transwell chamber were used to measure changes in PTC proliferative capacity and invasive ability.Results:In this study, we found that mBmi-1 expression was positively correlated with PTC lesion size and invasion ability. Bioinformatics prediction results suggested that Bmi-1 was a target gene of microRNA-203 (MicroRNA-203, miR-203) , and the 3’-UTR target sequence of miR-203 against Bmi-1 was GUAAAGU. After transfection of the miR-203 mimetic, the expression of Bmi-1 mRNA in the PTC cell line TPC-1 was significantly down-regulated. The dual luciferase reporter gene assay demonstrated that miR-203 can act on the 3’UTR region of the Bmi-1 gene to predict the target. In addition, the transfection of miR-203 mimetic effectively inhibited the proliferation and invasion of PTC cells, and the ability of proliferation and invasion of PTC cell TPC-1 was restored after overexpression of Bmi-1 in PTC cell TPC-1 transfected with miR-203 mimic.Conclusions:Bmi-1 is positively correlated with the proliferation and invasion ability of PTC. miR-203 can inhibit the proliferation and invasion of PTC cells by directly targeting Bmi-1 gene.