Objective To investigate the effects of trivalent arsenicals on ce ll proliferation and induction of apoptosis in human epidermal keratinocytes. Me thods Human benign epidermal keratinocytes (cell line HaCaT), human epidermal ca rcinoma cells(cell line A431) were cultured. After treatment with arsenous acid, inhibition of cellular growth was determined by measuring MTT dye absorption of living cells.Apoptosis was assessed with respect to morphological changes by li ght and electron microscopy and to cell cycle distribution by flow cytometry. An nexin-ⅴbinding assay was used to detect the early stage of apoptosis. Results With concentrations ranging from 0.5 to 10 ?mol/L, arsenous acid significantly inhibited the proliferation of HaCaT cells in a dose-and time-dependent manner . By light and electron microscopy, morphological changes revealed characteristi cs of apoptosis. But A431 cells showed no obvious change. DNA flow cytometric an alysis indicated that arsenous acid induced an arrest in G2M phase and sub-G1 p hase in HaCaT compared with A431 cells. The green flurorescence indicated early stage of apoptosis in HaCaT cells by annexin-V binding assay. Conclusion Arseno us acid may inhibit the proliferation of HaCaT cells and induce apoptosis, but d oes not affect A431 cell line obviously, which suggests that HaCaT cells are mor e sensitive to arsenous acid compared with A431 cells.

Objective To observe the effect of ethanol extract and ethyl acetate extract of Capparis Spinosa on the thickness of dermis,synthesis of collagen type Ⅰ,type Ⅲ,and expression of transforming growth factor-β1 in mouse models of scleroderma.Methods Mouse models of scleroderma were established through local injection of bleomycin on the back once a day for 4 weeks.After confirmation of model establishment,72 mouse models were equally and randomly divided into three groups.Two groups received topical treatment with ethanol extract of Capparis Spinosa and ethyl acetate extract of Capparis Spinosa,respectively,no treatment was given to the rest of the control group.After 2-,4-,6-week treatment,8 mice were sacrificed and tissue samples were obtained from the back,and subiected to the measurement of dermal thickness by HE staining,as well as to the analysis of expression of collagen type Ⅰ,collagen type Ⅲ and transforming growth factor-β1 by immunohistochemical staining.Results On week 2,4,6,the thickness of dermis was 23.22,24.94,19.97 μm respectively in mice treated with ethanol extract of Capparis Spinosa,27.66.26.15,22.13 μm respectively in those treated with ethyl acetate extract of Capparis Spinosa.Compared with the mouse models without treatment,the thickness of dermis significantly decreased(F=12.99,P＜0.01),the expression of collagen type Ⅰ(F=7.47,P＜0.01)and transforming growth factor-β1(F=11.76,P＜0.01)were also inhibited in those receiving treatment.However,the expression of collagen type Ⅲ was not affected obviously by the treatment.Conclusion The ethanol extract and ethyl acetate extract of Capparis Spinosa have the effect against skin fibrosis.

0.1). Plasma PAF-AH activity in the patients with psoriasis was significantly lower than that in the healthy controls(P

Posttranslational modifications of antibody products affect their stability,charge distribution,and drug activity and are thus a critical quality attribute.The comprehensive mapping of antibody modifications and different charge isomers(CIs)is of utmost importance,but is challenging.We intended to quanti-tatively characterize the posttranslational modification status of CIs of antibody drugs and explore the impact of posttranslational modifications on charge heterogeneity.The CIs of antibodies were fraction-ated by strong cation exchange chromatography and verified by capillary isoelectric focusing-whole column imaging detection,followed by stepwise structural characterization at three levels.First,the differences between CIs were explored at the intact protein level using a top-down mass spectrometry approach;this showed differences in glycoforms and deamidation status.Second,at the peptide level,common modifications of oxidation,deamidation,and glycosylation were identified.Peptide mapping showed nonuniform deamidation and glycoform distribution among CIs.In total,10 N-glycoforms were detected by peptide mapping.Finally,an in-depth analysis of glycan variants of CIs was performed through the detection of enriched glycopeptides.Qualitative and quantitative analyses demonstrated the dynamics of 24 N-glycoforms.The results revealed that sialic acid modification is a critical factor ac-counting for charge heterogeneity,which is otherwise missed in peptide mapping and intact molecular weight analyses.This study demonstrated the importance of the comprehensive analyses of antibody CIs and provides a reference method for the quality control of biopharmaceutical analysis.

Amino Acid Sequence ; Genes, Bacterial ; Hydrogen Peroxide ; pharmacology ; Molecular Sequence Data ; Mutation ; genetics ; Mycobacterium ; drug effects ; genetics ; growth & development ; Peptides ; chemistry ; metabolism ; Stress, Physiological ; drug effects