目的:研究青稞多糖(HVP)对糖尿病模型小鼠的降血糖作用及机制.方法:对小鼠腹腔注射链脲佐菌素(120 mg/kg)以复制糖尿病模型.将造模成功的小鼠随机分为模型组、二甲双胍组(阳性对照,200 mg/kg)和HVP高、中、低剂量组(300、150、75 mg/kg),另设空白对照组,每组10只.各给药组灌胃相应药物,空白对照组和模型组小鼠灌胃等量水,每天1次,连续30天.给药10、20、30天时测定小鼠的空腹血糖(FBG);末次FBG测定后,小鼠腹腔注射10％葡萄糖溶液(2 g/kg),于注射后30、120 min时测定糖耐受量(GTT)曲线下面积;测定小鼠血清中胰岛素水平和肝组织中超氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GSH-Px)、丙二醛(MDA)的含量;采用苏木精-伊红染色法观察小鼠胰腺组织病理学变化.结果:与空白对照组比较,模型组小鼠FBG、GTT曲线下面积和肝组织中MDA含量均显著升高或增加(P<0.01);血清中胰岛素水平和肝组织中SOD、GSH-Px含量均显著降低(P<0.01);胰腺组织损伤严重,形态不完整,有明显的空泡.与模型组比较,HVP高剂量组小鼠FBG、GTT曲线下面积和肝组织中MDA含量均显著降低或减少(P<0.05或P<0.01),血清中胰岛素含量和肝组织中SOD、GSH-Px含量均显著升高(P<0.05或P<0.01),HVP低、中剂量组上述指标部分改善,差异具有统计学意义(P<0.05或P<0.01);胰腺组织损伤有所减轻,形态较为完整,胰岛细胞排列紧密,空泡减少.结论:HVP可降低糖尿病模型小鼠的血糖水平,升高胰岛素水平并减轻胰腺组织损伤;该作用机制可能与其抗氧化作用有关.

Polysaccharide from traditional Chinese herb, Saposhnikovia divaricata (Turcz.) Schischk. (SD) was extracted, fractionated and characterized in this work. Four fractions were prepared. Their molecular weight, monosaccharide compositions, linkage modes and structural properties were characterized with SEC-MALS-RI, HPAEC-PAD, GC-MS and NMR. SDP1 was assigned as a 1, 4-α-glucan with small amount of O-6 linked branches. SDP2 contained a big amount of the 1, 4-α-glucan and a small amount of arabinogalactan, while SDP3 possessed relatively lower amount of the 1, 4-α-glucan and a big amount of the arabinogalactan. SDP4 was defined as a pectic arabinogalactan. Four fractions showed antioxidant activities in both molecular and cellular levels and their activity was ranked as SDP4 ≈ SDP3>SDP2>SDP1. The 1, 4-α-glucan in SDP1 had the weakest, while SDP3 and SDP4 showed similar and the highest antioxidant activity. The arabinogalactan was the major component of both SDP3 and SDP4, which significantly contributed to the antioxidant activity of SDP.

目的:研究臭灵丹对人白血病K562和NB4细胞的抑制作用及其黄酮类成分.方法:应用四甲基偶氮噻唑蓝比色法(MTT法)测臭灵丹对K562和NB4细胞生长的抑制率,采用PI/Annexin-V双染色方法考察臭灵丹对K562和NB4的凋亡作用;运用大孔树脂、Sephadex LH-20柱色谱进行化学成分的分离纯化,通过波谱分析鉴定化合物结构.结果:臭灵丹对K562和NB4细胞的抑制作用呈现剂量依赖性;臭灵丹能剂量依赖性地诱导K562和NB4细胞的凋亡;从臭灵丹中分离得到了6个黄酮类化合物,分别鉴定为洋艾素、金腰乙素、5-羟基-3',4',7-三甲氧基黄酮、雷杜辛黄酮醇、芹菜素-7-O-β-D-葡糖糖苷、槲皮素.结论:臭灵丹具有抗K562和NB4细胞的作用,首次从臭灵丹中分离得到雷杜辛黄酮醇.

Hyaluronic acid (HA), a glycosaminoglycan, is a linear polysaccharide with negative charge, composed of a repeating disaccharide unit [→4)-β-d-glucopyranosyluronic acid (1→3)-β-d- N-acetyl-d-glucoaminopyranose (1→]n ([→4) GlcA (1→3) GlcNAc 1→]n). It is widely used in different applications based on its physicochemical properties associated with its molecular weight. Enzymatic digestion by polysaccharides lyases is one of the most important ways to decrease the molecular weight of HA. Thus, it is important to understand the action patterns of lyases acting on HA. In this study, the action patterns of two common lyases, Flavobacterial chondroitinase AC I and Streptomyces hyaluronidase, were investigated by analyzing HA oligosaccharide digestion products. HA oligosaccharides having an odd-number of saccharide residues were observed in the products of both lyases, but their distributions were quite different. Chondroitinase AC acted more efficiently at the β 1-4 glycosidic bond linking GlcNAc and GlcA. Oligosaccharides, having an even number of saccharide residues, and with an unsaturated uronic acid (4-deoxy-α-l-threo-hex-4-enepyranosyluronic acid, △UA) residue at their non-reducing end represent the major product. A minor amount of oligosaccharides having an odd number of saccharide residues resulted from the irregular terminal residues of HA substrate chains. Hyaluronidase showed a more complicated product mixture. Its minimum recognition and digestion domain is HA heptasaccharide and it could cleave both β 1-4 and β 1-3 glycosidic linkages. The HA oligosaccharides, generated with a 2-acetamido-2,3-di-deoxy-β-d-erythro-hex-2-enopyranose (△HexNAc) at the non-reducing end, are believed to be unstable and undergo breakdown immediately after their generation, and the oligosaccharides with △UA residue at the non-reducing end are formed. Thus, oligosaccharides having both an even and odd-number saccharide residues with a △UA residue at their non-reducing ends, represent the major products of hyaluronidase acting on HA.

Highland barley is one of the hull-less barleys that contains high amounts of beta-glucan. A detailed sequence analysis is needed to support the application of the beta-glucan from highland barley. The beta-conformation and 1 -> 3/1 -> 4 linkages were confirmed using nuclear magnetic resonance (NMR) spectroscopy. Monosaccharide analysis was accomplished with high-performance anion exchange chromatography (HPAEC) coupled to pulsed amperometric detection (PAD) and showed that glucose (G) was the dominant sugar component of this b-glucan. HPAEC-PAD was applied with on-line electrospray ion - to quadrupole time - of - flight mass spectrometry (Q/TOF-MS) to sequence oligosaccharides enzymatically derived from this beta-glucan. More than 20 oligosaccharides were observed in the partially digested beta-glucan mixture and their degree of polymerization (dp), confirmed by MS analysis, ranged from dp 2 to dp 20. The sequences of oligosaccharide dp2 - dp7 in the digested product mixture were unambiguously determined with MS/MS. Based on the sequence analysis of these oligosaccharides, G1 -> 3G1 -> 4G and G1 -> 3G1 -> 4G1 -> 4G were determined to be the major repeating blocks of this beta-glucan, and they were interspersed with homogeneous 1 -> 4 linked domains with different lengths. The ratio of 1 -> 3 and 1 -> 4 linkages in the beta-glucan was approximately 1:3. (C) 2017 Elsevier Ltd. All rights reserved.

Objective: To investigate the effect of obstructive sleep apnea (OSA) on sleep architecture in acute ischemic stroke (AIS) patients. Methods: Seventy AIS patients with polysomnography examination from June 2014 to April 2016 were included in the Second Affiliated Hospital of Soochow University. Twenty-seven healthy controls during the same period were chosen as control group. According to apnea-hypopnea index (AHI), AIS patients were divided into AIS group (AHI<5/h) and AIS+ OSA group (AHI≥5/h). Their general conditions, Neurological function defect (national institutes of health stroke scale, NIHSS) on admission and at discharge and self-care ability (modified Rankin Scale, mRS) at discharge and 3 months later (mRS score <2 reflected good prognosis, and mRS score≥2 reflected unfavorable prognosis) were collected. Results: Fifty-two patients (74.3%) had OSA in total seventy AIS patients. Total sleep time (TST) was significantly shorter and sleep efficiency (SE) was lower in AIS group than the control group (P=0.007, 0.008, respectively). AIS+ OSA group had longer non-rapid eye movement (NREM)1 than control group [24.9(21.3) vs 14.3(10.6), P=0.044]. Compared with AIS group, AIS+ OSA group had shorter NREM3 [13.0(13.2) vs 19.6(12.8), P=0.039]. There was no significant difference between the infarct location of AIS group and AIS+ OSA group. However, AIS+ OSA group had higher mRS score observed at 3 months through follow-up visit than AIS group (P=0.027). Spearman correlation analysis showed a positive correlation between unfavorable prognosis of stroke at 3 months and atrial fibrillation, the oxygen desaturation index (ODI), percentage of oxygen saturation <90% of total recording time (TS90%), AHI, the longest time of apnea and hypopnea, respiratory related microarousal and NIHSS, while a negative correlation between unfavorable prognosis of stroke and NREM3, lowest nocturnal oxygen saturation (L-SaO(2)). On Logistic analysis, the unfavorable prognosis was independently predicted by AHI and NIHSS. Conclusions: Sleep architecture of cerebral infarction patients are disturbed with its characteristic of shorter total sleep time and lower sleep efficiency. Cerebral infarction patients with OSA have longer NREM1 and shorter NREM3.

Gnaphalium hypoleucum DC is used as a wild vegetable, named “Qing Ming Cai”, as well as a folk medicine in China. One new compound, Gnaphaliin C, and nineteen known compounds were isolated and indentified from G. hypoleucum, total flavonoids were analyzed by UHPLC-Q-TOF/MS. The α-glucosidase inhibitory and anti-complementary activities of all compounds and total flavonoids were evaluated. Total flavonoids and ten compounds exhibited anti-complementary activities. Total flavonoids and six compounds inhibited α-glucosidase. The antihyperglycaemic effects of total flavonoids were investigated by streptozotocin (STZ)-induced diabetic mice. Total flavonoids improved the glucose tolerance and decreased blood sugar, increased muscle glycogen content, decreased the levels of triacylglycerol (TG) and total cholesterol (TC), enhanced superoxide dismutase (SOD) activity and decreased malondialdehyde (MDA) level in diabetic mice. G. hypoleucum and its total flavonoids could be used as functional food for diabetics.

The glycosylation of proteins is responsible for their structural and functional roles in many cellular activities. This work describes a strategy that combines an efficient release, labeling and liquid chromatography-mass spectral analysis with the use of a comprehensive database to analyze N-glycans. The analytical method described relies on a recently commercialized kit in which quick deglycosylation is followed by rapid labeling and cleanup of labeled glycans. This greatly improves the separation, mass spectrometry (MS) analysis and fluorescence detection of N-glycans. A hypothetical database, constructed using GlycResoft, provides all compositional possibilities of N-glycans based on the common sugar residues found in N-glycans. In the initial version this database contains >8,700 N-glycans, and is compatible with MS instrument software and expandable. N-glycans from four different well-studied glycoproteins were analyzed by this strategy. The results provided much more accurate and comprehensive data than had been previously reported. This strategy was then used to analyze the N-glycans present on the membrane glycoproteins of gastric carcinoma cells with different degrees of differentiation. Accurate and comprehensive N-glycan data from those cells was obtained efficiently and their differences compared corresponding to their differentiation states. Thus, the novel strategy developed greatly improves accuracy, efficiency and comprehensiveness of N-glycan analysis.

Oxidized starch, one of the most important starch derivatives, has many different properties and applications. Currently, there are two ways to produce oxidized starch, through specific and nonspecific oxidation. Specific oxidation using the stable nitroxyl radical, 2,2,6,6-tetramethyl preparidinloxy (TEMPO), with NaBr and NaClO can produce oxidized starches with different properties under good quality control. In the current study, we examine the products of specifically oxidized starch. As the amount of oxidant and the temperature, two critical factors impacting the oxidation of starch were thoroughly investigated. Analysis of the molecular weight (MW), degree of oxidization (DO) and the detailed structures of corresponding products was accomplished using gel permeation chromatography with multi-angle laser light scattering (GPC-MALLS), infrared (IR) spectroscopy, nuclear magnetic resonance (NMR) spectroscopy, and quadrapole time-of-flight mass spectrometry (Q/TOF-MS). According to the analytical results, the oxidation patterns of starch treated with specific oxidant TEMPO-NaBr-NaClO were established. When high amounts of oxidant was applied, more glucose residues within starch were oxidized to glucuronic acids (higher DO) and substantial degradation to starch oligosaccharides was observed. By selecting a reaction temperature of 25°C a high DO could be obtained for a given amount of oxidant. The reducing end sugar residue within oxidized starch was itself oxidized and ring opened in all TEMPO-NaBr-NaClO reactions. Furthermore, extra oxidant generated additional novel structures in the reducing end residues of some products, particularly in low temperature reactions.

Ethnopharmacological relevance: Eupatorium lindleyanum DC. is widely used for its efficiency in treating cough, tracheitis and tonsillitis. Acute lung injury (ALI) induced by lipopolysaccharide (LPS) in mice was used to investigate therapeutic effects and possible mechanism of the sesquiterpenes fraction of E. lindleyanum DC. (EUP-SQT).Materials and methods: Mice were orally administrated with EUP-SQT (15, 30 and 60 mg/kg) per day for 7 days consecutively before LPS challenge. The lung specimens and bronchoalveolar lavage fluid (BALF) were harvested for histopathological examinations and biochemical analysis at 6 h and 24 h after LPS challenge. The level of complement 3 (C3) and complement 3c (C3c) in serum was quantified by a sandwich ELISA kit.Results: Pretreatment with EUP-SQT could significantly decrease lung wet-to-dry weight (W/D) ratio, nitric oxide (NO) and protein concentration in BALF, which was exhibited together with the lowered myeloperoxidase (MPO) activity, the increased superoxide dismutase (SOD) activity and down-regulation the level of tumor necrosis factor-alpha (TNF-alpha), interleukin-6 (IL-6) and interleukin-1 beta (IL-1 beta) in ALI model. Additionally, EUP-SQT attenuated lung histopathological changes and significantly reduced complement deposition with decreasing the level of C3 and C3c in serum.Conclusions: These results showed that EUP-SQT significantly attenuated LPS-induced ALI via reducing productions of pro-inflammatory mediators and decreasing the level of complement, indicating it as a potential therapeutic agent for ALI. (C) 2016 Elsevier Ireland Ltd. All rights reserved.

Microcrystalline cellulose, a major component of cell wall of plants, is one of the most abundant natural materials, but the poor solubility of cellulose limits its applications. Cellulose is a linear glucan with exclusive beta 1 -> 4 linkage. Oxidation carried out with TEMPO-NaBr-NaClO system can selectively oxidize the C6 of glucose residues in cellulose. This modification improves polysaccharide solubility and other physicochemical properties. In this work, the impact of degree of oxidation on solubility, degree of crystallization, thermostability, molecular weight and the structures of the resulting oligosaccharide products of selectively oxidized cellulose were investigated using x-ray diffraction, thermogravimetric analysis, gel permeation chromatography-multiple angle laser light scattering and ultrahigh performance liquid chromatography-electrospray-quadrupole/time of flight-mass spectrometry, respectively. The physicochemical properties of selectively oxidized cellulose having different degrees of oxidation were carefully characterized providing a theoretical foundation for the more accurate selection of applications of oxidized celluloses. (C) 2016 Elsevier Ltd. All rights reserved.

中药化学实验课的教学在中药专业中占据很重要的地位。我们在实验教学中，“以学生为中心”，采用先进的教学内容和科学的教学方法，积极引导学生的创造性思维，培养学生自主学习，以科研促进教学，开展学生与教师之间的良性互动，取得较好的教学效果。

药物分析实验教学以提高学生的实践能力、应用能力、创新能力和综合素质为目标,在教学改革和教学理念不断更新的背景下,要求教师实践密切联系理论之外,更要激发学生的学习兴趣,注重学生实践与探究能力的培养.在药物分析实验教学中运用案例教学法,结合自主探究性学习模式,不仅激发学生的学习积极性,拓宽学生知识面和视野,也为药学教育者正确、合理运用案例教学法提供参考.

翻转课堂作为一种新型的教学模式，将该模式引入到药物分析课程的教学中，运用信息化和数字化教学资源及手段，分析传统教学方法存在的问题，设计教学案例，变革课堂组织形式，过程分为知识探索阶段、吸收内化阶段及点评思考阶段，设计“学生为中心，教师为主导，训练为主线”的翻转课堂教学模式，可以改变传统课堂教师讲、学生听的教学形式，能有效提高学生的学习积极性和学习效率，更能提升学生自主学习、课后反思、解决问题的能力，实现教学评价的多元化与智能化。

Investigation of the action pattern of enzymes acting on carbohydrates is challenging, as both the substrate and the digestion products are complex mixtures. Dextran and its enzyme-derived oligosaccharides are widely used for many industrial applications. In this work, a new method relying on ultra-performance hydrophilic interaction liquid chromatography quadrupole time-of-flight tandem mass spectrometry (UP-HILIC-Q/TOF-MS/MS) was developed to analyze a complex mixture of dextran oligosaccharide products to determine the action pattern of dextranase. No derivatization of oligosaccharides was required and the impact of the α- and β-configurations of the native oligosaccharides on the chromatographic separation was eliminated. The 1→6, 1→3, 1→4 backbone linkages and the branch linkages of these oligosaccharides were all distinguished from diagnostic ions in their MS/MS spectra, including fragments corresponding to (0,2)A, (0,3)A, (0,4)A, B-H2O, (2,5)A, and (3,5)A. The sequences of the oligosaccharide products were similarly established. Thus, the complex oligosaccharide mixtures in dextran digestion products were profiled and identified using this method. The more enzyme-resistant structures in dextran were established using much less sample, labor, time, and uncertainty than in previous studies. This method provides an efficient, sensitive, and straightforward way to monitor the entire process of digestion, establish the action pattern of the dextranase from Penicillium sp., and to support the proper industrial application of dextranase.

PBL教学法可以促进学生进行开放式学习，在中药注射剂质量分析教学中引入PBL教学法，可提升学生思维能力、信息检索能力以及交流能力。 PBL教学不仅使学生加深对课程的认识，而且真正实现以人为本的教育核心理念。

Alginate is a linear and acidic polysaccharide, composed of (1 → 4) linked β-D-mannuronic acid (ManA) and α-L-guluronic acid (GulA). The ratio of ManA to GulA (M/G) is one of the most important factors for the application of alginate and its derivatives in various areas. In this work, a robust and accurate method was developed to analyze M/G using high-performance anion-exchange chromatography with pulsed amperometric detection (HPAEC-PAD). The impact of hydrolysis conditions on the release patterns of ManA and GulA from alginate and its derivatives was investigated. The release patterns of ManA and GulA need to be considered separately to obtain an accurate M/G. Several hydrolysis conditions were established that released ManA and GulA completely and maintained these saccharide residues intact. The proper M/G of alginates from different sources and its derivatives could then be calculated by integration of the corresponding ManA and GulA peaks.