Vitiligo is a stubborn multifactorial skin disease with a prevalence of approximately 1% in the global population. Kaliziri, the seeds of Vernonia anthelmintica (L.) Willd., is a well-known traditional Uyghur medicine for the treatment of vitiligo. Kaliziri injections is a Chinese-marketed treatment approved by the China Food and Drug Administration for the treatment of vitiligo. The significant effects of Kaliziri injection have been thoroughly studied. However, chemical components studies and plasma quantification studies are lacking for Kaliziri injection. Ultra-high-performance liquid chromatography coupled with hybrid quadrupole orbitrap mass spectrometry was employed to comprehensively characterize the caffeoyl quinic acid derivatives present in Kaliziri injection. Based on accurate mass measurements, key fragmental ions and comparisons with reference standards, 60 caffeoyl quinic acid derivatives were identified in Kaliziri injections, including caffeoyl quinic acids, coumaroyl caffeoyl quinic acids, dicaffeoyl quinic acids, feruloyl caffeoyl quinic acids, and dicaffeoyl quinic acid hexosides. Moreover, an HPLC-MS/MS method was developed and validated for the quantitative analysis of 5-caffeoyl quinic acid, 4-caffeoyl quinic acid, 1,3-dicaffeoyl quinic acid, 3,4-dicaffeoyl quinic acid, 3,5-dicaffeoyl quinic acid and 4,5-dicaffeoyl quinic acid in beagle plasma. The quantitative HPLC-MS/MS method was applied to quantify these six major caffeoyl quinic acids in beagle plasma after the subcutaneous administration of Kaliziri injection. All of the six analytes reached their peak plasma of concentrations within 30 min.

In this study, an enzyme-assisted extraction method was used to extract Orchis chusua D. Don (Salep) polysaccharide (SP), which was then modified by sulfation, acetylation, phosphorylation, and carboxymethylation to obtain modified polysaccharides. Furthermore, their degree of substitution, chemical composition, and molecular weight were evaluated. The primary structural features were characterized by UV spectra, FT-IR spectra, Congo-red test, and scanning electron microscope. The phosphorylated polysaccharide (SP-P) was demonstrated the highest scavenging ability on hydroxyl radical and growth-promoting activity on Lactobacillus Bulgaricus. The carboxymethylated polysaccharide (SP-C) was exhibited the strongest DPPH and ABTS radical scavenging effects. The acetylated polysaccharide (SP-A) displayed the best proliferation effects on Bifidobacterium adolescentis, whereas the sulfated polysaccharide (SP-S) maintained moderately stable antioxidant and probiotic ability. These findings indicate that the modified polysaccharides had their potential significance as new antioxidants and probiotics for the food industry. PRACTICAL APPLICATION: This article provides a new source for the development of polysaccharide derivatives as new antioxidants and probiotics for the food industry.

As an active compound, psoralen is present in various Chinese herbal medicines and has exhibited significant activity in skin disease treatment. Its derivative 8-methoxypsoralan (8-MOP) is the most commonly used drug to induce repigmentation of vitiligo. In our previous screening assays, 4-methyl-6-phenyl-2H-furo[3,2-g]chromen-2-one (MPFC), a psoralen derivative, was identified as more effective tyrosinase and melanin activator than the positive control 8-MOP in consideration of low doses, as well as low toxicity. The overall purpose of this study was to characterize the melanogenic effect and mechanisms of MPFC in B16 cells. The melanin biosynthesis effects of MPFC were determined by examination of cellular melanin contents, tyrosinase activity assay, cyclic adenosinemonophosphate (cAMP) assay, and western blotting of MPFC-stimulated B16 mouse melanoma cells. Our results showed that MPFC enhanced both melanin synthesis and tyrosinase activity in a concentration-dependent manner as well as significantly activated the expression of melanogenic proteins such as tyrosinase, tyrosinase-related protein-1 and tyrosinase-related protein-2. Western blot analysis showed that MPFC increased the phosphorylation of p38 mitogen-activated protein kinase and cAMP response element-binding protein (CREB) as well as the expression of microphthalmia-associated transcription factor (MITF). Moreover, MPFC stimulated intracellular cAMP levels and induced tyrosinase activity and melanin synthesis were attenuated by H89, a protein kinase A inhibitor. These results indicated that MPFC-mediated activation of the p38 MAPK and the protein kinase A (PKA) pathway may shed light on a novel approach for an effective therapy for vitiligo.

Plants or plant-derived products have been routinely used in several traditional medicine systems for vitiligo treatment. It is well-known that melanogenesis can be promoted by certain flavonoid compounds isolated from the traditional Uyghur medicinal plant, Kaliziri. Therefore, Chalcones, one class of flavonoid compounds, has become an interesting target for the development of anti-vitiligo agents. A series of novel isoxazole chalcone derivatives have been designed, synthesized, and evaluated for biological activities by our group. Among them, derivative 1-(4-((3-phenylisoxazol-5-yl)methoxy)phenyl)-3-phenylprop-2-en-1-one (PMPP) was identified as a potent tyrosinase activator with better activity and lower toxicity than the positive control 8-methoxypsoralen (8-MOP) in this study. Further investigations revealed that Akt and GSK3β were the signaling pathways involved in the hyperpigmentation of PMPP. Overall, these studies may provide a convenient and novel approach for the further development of anti-vitiligo agents.

Isochlorogenic acid A, also called 3,5-dicaffeoylquinic acid (3,5-diCQA), is a widespread phenolic compound in the plant. Recent studies have shown that it has antioxidant and anti-inflammatory activity. In addition, oxidative stress and inflammation induced by solar ultraviolet radiation is a very significant reason for skin depigmentation. Therefore, in this study, we evaluated the effect of 3,5-diCQA on B16 cells and explored its molecular mechanism. Results showed that 3,5-diCQA upregulated intracellular melanin production in a time- and dose-dependent manner. Tyrosinase (TYR) activity was also increased after treatment with 3,5-diCQA in a dose-dependent manner. Expressions of TYR, TYR-related protein1, TYR-related protein2, and microphthalmia-associated transcription factor were upregulated in a dose-dependent manner after 48 h of treatment with 3,5-diCQA. Results also showed that 3,5-diCQA promoted the phosphorylation of Akt at Thr308 and glycogen synthase kinase-3β at Ser 9. Moreover, 3,5-diCQA increased the content of β-catenin in cell cytoplasm and nucleus by reducing the content of phosphorylated β-catenin (p-β-catenin). All these results suggest that 3,5-diCQA may mediate the acceleration of melanin synthesis by the β-catenin signal pathway.

A new series of chalcone derivatives 1–18, bearing isoxazole moieties were designed and synthesized, and biologically evaluated for their activity on mushroom tyrosinase and melanin synthesis in murine B16 cells. The result indicated that most of prepared compounds 1–18 showed potent activating effect on tyrosinase, especially for 1–2, 4, 6–7, 9 and 15. Among them, compounds 2, 4 and 9 demonstrated the best activity with EC50 = 1.3, 2.5 and 3.0 μmol·L−1 respectively, much better than the positive control 8-methoxypsoralan (8-MOP, EC50 = 14.8 μmol·L−1); In B16 cells, all the tested compounds exhibited a stronger activity on melanogenesis than 8-MOP (with the value of 115%). It was interesting that derivatives substituted with halogen (1, 2, 4, 5, 7, 9) were generally more potent. Compounds 2 (463%) and 18 (438%) with 3 and 4-fold potency compared with 8-MOP respectively, were recognized as the most promising candidate hits for further pharmacological study of anti-vitiligo.

Furocoumarins, isolated from Psoralen corylifolia L., were found to be the most effective drug in the treatment of vitiligo nowadays. Twenty-five furocoumarin derivatives were thus designed and synthesized in order to improve the melanogenesis in B16 cells for the first time. Among them, twenty-three compounds were more potent than the positive control (8-MOP), the commonly used drug for vitiligo in clinic. Noticeably, compounds 6m (350.5%) and 6p (313.1%) based on the scaffold of 6k (2H-benzofuro[2,3-h] chromen-2-one) were nearly 3-fold stronger than 8-MOP (114.50%). The in vitro melanin synthesis evaluation of these structurally diverse analogues had also led to an outline of structure-activity relationship. (C) 2016 Elsevier Ltd. All rights reserved.

目的：评价玫瑰花提取物的体外降糖活性，研究玫瑰花多酚作用的胰岛素信号通路并阐明其降糖作用机制。方法采用脂质体介导的质粒转染法构建蛋白酪氨酸磷酸酶-1B( PTP1B)蛋白过表达的CHO-K1细胞模型，通过蛋白印迹分析法( Western blotting)研究玫瑰花多酚对细胞内信号分子蛋白表达量的影响。结果体外玫瑰花提取物抑制PTP1B酶活性，半数抑制浓度( IC50)为62.31 ng.mL－1。玫瑰花提取物可提高细胞葡萄糖消耗，与阳性对照药作用相当；在细胞内可明显提高磷酸化的胰岛素受体底物1( IRS-1)、磷脂酰肌醇依赖性激酶1( PDK1)、蛋白激酶B( AKT)以及糖原合成激酶3β( GSK-3β)蛋白表达量。结论玫瑰花提取物对PTP1B有良好的抑制活性，可促进葡萄糖消耗，具有潜在降糖活性；玫瑰花提取物通过增加IRS-1、下游信号分子PDK1、AKT以及GSK-3β蛋白的磷酸化水平来激活PI3K/AKT胰岛素信号通路，从而促进胰岛素信号传导和糖原合成，达到降低血糖的效果。

Dipeptidyl peptidase IV(DPP IV) is a new target for treating type II diabetes. Glucose-dependent insulin-releasing peptide, and the insulin glucagon-like peptide-1 could promote insulin secretion, biosynthesis, inhibit β-cell apoptosis, lower the blood sugar and other functions. DPP IV could degradate the level of glucose-dependent insulin-releasing peptide and glucagon-like peptide-1 to the loss of biological activity. Our experiment is to optimize the screening system for DPP IV inhibitors, based on DPP IV from Caco-2, results showed that the concentration of Gly-Pro-pNA is 0.064 mg/mL,reaction time is 30 minutes, temperature 37 ?C and the concentration of IIe-Pro-IIe 0.68 mmol/L. Therely astablishing a rapid screening system of dipeptidyl peptidase. Application of the model 18 Uyghur herbal extracts were screened and some had the inhibitory activity of DPP IV. The study will provide support for further exploration and utilization of Uyghur herbs.

Our previous research showed that standardized extract from the flowers of the Gossypium herbaceam labeled GHE had been used in clinical trials for its beneficial effects on brain functions, particularly in connection with age-related dementia and Alzheimer's disease (AD). The aim of this work was to determine the components of this herb and the individual constituents of GHE. In order to better understand this herb for AD treatment, we investigated the acetylcholinesterase (AChE) inhibition and antioxidant activity of GHE as well as the protective effects to PC12 cells against cytotoxicity induced by tertiary butyl hydroperoxide (tBHP) using in vitro assays. The antioxidant activities were assessed by measuring their capabilities for scavenging 1,1-diphenyl-2-picylhydrazyl (DPPH) and 2-2'-azinobis-(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) free radical as well as in inhibiting lipid peroxidation. Our data showed that GHE exhibited certain activities against AChE and also is an efficient free radical scavenger, which may be helpful in preventing or alleviating patients suffering from AD.

The present study aimed to examine the chemical composition and biological activity of essential oils extracted from Euphorbia macrorrhiza collected from Northwest China. The major constituents of the essential oils of aerial parts and roots of E. macrorrhiza are acorenone B (16.72% and 25.80%), (+)-cycloisosativene (14.94% and 12.40%), 3a-hydroxy-5b-androstane (10.62% and 5.52%), copaene (7.37% and 6.29%), l-calamenene (4.13% and 4.65%) and β-cedrene (8.40% and 7.98%), respectively. The minor components of them are thymene, γ-terpinene, thymecamphor, α-cedrene, zingiberene, trans-caryophyllene, β-chamigrene, curcumene, pentadecane, (−)-α-muurolene, cuparene, γ-cadinene, (Z)-3-heptadecene, 1,3,7,7-tetramethyl-2-oxabicyclo(4.4.0)dec-5-en-4-one, hexahydrofarnesyl acetone, γ-elixene and palmitinic acid. The antimicrobial and antitumor activitiy of the E. macrorrhiza essential oil against Staphyloccocus aureus, Escherichia coli, Canidia Albicans and Caco-2 cells were evaluated. Among all the tested microorganisms and Caco-2 cells, the essential oils showed the strongest inhibitory effect on Staphyloccocus aureus (MIC = 2.8 μg/mL) and Caco-2 cell (IC50 = 11.86 μg/mL), whereas no effect on Escherichia coli and Candida albicans. The data of this study suggested that the E. macrorrhiza essential oils have great potential as a natural medicine for microbial infections and cancers.

This paper aimed to explore inhibitory effectof extracts from Astragalus altaicus Bge on the protein tyrosine phosphatase1B.The contents of flavonoids and saponins of eight extracts(E1-8) obtained from A.altaicus were measured by spectrophotometric assay.The results indicated that contents of flavonoids of E1-8 were 5.09,10.46,3.58,3.23,53.91,21.77,5.76 and 7.49,respectively,and those of saponins of E1,E2,E6,E7 and E8 were 34.8,4.7,7.35 and 7.15 μg/mL,respectively.The in vitro PTP1B inhibitor screening system was applied to detect the inhibitory ability and the results indicated that IC50 of E1,E2,E7,and E8 were 34.8,4.7,7.35and 7.15 μg/mL,respectively.The enzyme kinetic studies revealed non-competitive type of inhibition of E1,E7,E8 and mixed competitive type of inhibition of E2.In addition,E1,E2,E5,E7 and E8 obviously enhanced the rate of glucose intake in CHO cells,but E3,E4 and E6 were not.Therefore saponins might be major active substances in A.altaicus,which could increase the rate of glucose intake in CHO cells by PTP1B pathway.These findings might provide experimental support for further exploration and utilization of A.altaicus.

目的利用酶标仪,通过DPPH方法对草棉花不同提取物的抗氧化活性进行研究。方法以VC为对照I,C50值作为评价试样清除自由基能力为指标,建立草棉花提取物抗氧化活性快速筛选的方法。结果通过比较草棉花不同提取物对自由基的清除作用,经大孔树脂处理富集所得的总黄酮部位抗氧化活性最好。结论该实验为草棉花总黄酮的应用及药理研究提供理论依据。

Protein tyrosine phosphatase-1B is one of new drug targets against insulin sensitizer.In this study,we investigated the inhibitory effect of petroleumether extracts from 3 Uygur herbs,including Ziziphora Clinopodioides Lam.,Helichrysum arenarium(Linn.) Moench,and Ver-nohia anthelmintica(L.) willd,on the PTP1B activity and the mode of inhibition.The results showed that recombinant PTP1B was highly expressed by the prokaryotic expression system and its molecular weigh was 40.8 kDa.Three extracts showed different inhibitory effects on the PTP1B activity,with IC50 values of 20.2 μg/mL,12.65 μg/mL,and 9.7 μg/mL,respectively.The enzyme kinetic studies revealed non-competitive type of inhibition,competitive type of inhibition,and mixed competitive type of inhibition with the inhibitory constants(Ki) of 0.275 μg/mL,1.245 μg/mL,and 0.192 μg/mL,respectively.The study will provide support for further exploration and utilization of Uygur herbs.

Objective To explore the effects of chickpea sprout ethanol extract(CSE)on the proliferation and apoptosis of human colon adenocarcinoma cell line Caco-2 in vitro.Methods The active elements of chickpea sprout-CSE,were extracted and its components were analyzed.The proliferation inhibition of CSE on Caco-2 cells was measured by MTT assay.The ultrastructure of these cells was observed under electron microscope.Apoptosis was assayed by agarose gel electrophoresis,and cell apoptosis rate was evaluated by flow cytometry.Results CSE contained 23.53% of saponins and 14.73% of isoflavones.It inhibited Caco-2 cell proliferation in a dose-and time-dependent manner.Caco-2 cells treated with CSE showed chromatin margination and a few apoptotic bodies under electron microscope.DNA electrophoresis displayed a gradient band,which indicated that CSE could induce the apoptosis of Caco-2 cells.After treatment with CSE of 3,5,10 μg/ml for 24 h,the apoptosis was 32.6%,68.8% and 73.9%,respectively(P<0.05).Conclusion CSE can significantly inhibit the proliferation of Caco-2 cells by inducing the apoptosis,which provides the rationale for the application of chickpea to prevention and cure of human colon adenocarcinoma.

The leaves,stems,trunks and roots of tomato contained many antioxidants,which were previously discarded.It was studied that vitamin C and total flavonoids content in leaves,stems,trunks and roots were still very high after fruit ripening,which could be extracted to provide additives for food and medicine so as to make an effective use of them and achieve industrialization.

An effective high-speed countercurrent chromatography (HSCCC) method was established for further separation and purification of four minor flavonols in addition to five major flavonols which were reported by our previous study from extracts of Flos Gossypii. HSCCC was performed with three two-phase solvent systems composed of n-hexane-ethyl acetate-methanol-water (7.5:15:6:7, v/v), (2.5:15:2:7, v/v) and (0:1:0:1, v/v). The separation was repeated 3 times, and 3.8 mg of 8-methoxyl-kaempferol-7-O-β-D-rhamnoside (HPLC purity 98.27%), 6.7 mg of astragalin (HPLC purity 94.18%), 3.3 mg of 4'-methoxyl-quercetin-7-O-β-D-glucoside (HPLC purity 94.30%) and 8.2 mg of hyperoside (HPLC purity 93.48%) were separated from 150 mg of the crude sample. The chemical structures of the flavonols were confirmed by MS, (1)H NMR and (13)C NMR. Meanwhile, the results indicated that the target compound with smaller K value (<0.5) can be separated by increasing column length of HSCCC. And four separation rules of flavonols according to the present study and references were summarized, which can be used as a useful guide for separation of flavonols by HSCCC.