Type 3 resistant starch (RS3) was developed from Canna edulis (Ce) native starch (NS) through dual enzymatic hydrolysis and recrystallization. Thereafter, the processed Ce-RS3 was subjected to systematic characterizations for its structural properties, anti-hyperlipidemic effect, and in vivo gut microbiota modulatory function. The Ce-RS3 content was increased to 49.11% after processing under optimal conditions. Compared with NS, Ce-RS3 maintained its B-type crystallization without introducing new chemical groups. Meanwhile, it displayed coarse surfaces, higher crystallinity, more ordered structures, and a higher proportion of chains with degree of polymerization (DP) 37-100. Ce-RS3 intervention significantly alleviated dyslipidemia in hyperlipidemic mice, which was associated with increased gut microbial diversity and unique microbial enrichment, potentially mediated by its fine structure. These observations are valuable for developing RS3 from C. edulis for prebiotics applications and support the potential strategy that utilizes well-designed RS to modulate specific bacterial populations to improve health.

Obesity is a most prevalent human health problem. Several studies showed that appropriate modulation of gut microbiota could help reshape the metabolic profile of obese individuals, thereby altering the development of obesity. A nutritional strategy for treating obesity includes prebiotics. Type 3 Resistant Starch from Canna edulis (Ce-RS3) is a dietary fiber that exerts potential effects on the intestinal microbial community; however, the metabolic landscape and anti-obesity mechanism remain unclear. In the present study, obese mice were treated with Ce-RS3, and 16S rRNA gene sequencing and metabolomics were used to measure changes in gut microbiota and fecal metabolic profiles, respectively. At the end of the treatment (13 weeks), we observed slow weight gain in the mice, and pathological damage and inflammation were substantially reduced. Ce-RS3 constructs a healthy gut microbiota structure and can enhance intestinal immunity and reduce metabolic inflammation. Ce-RS3 increased the diversity of gut microbiota with enrichment of Bifidobacterium and Roseburia. Ce-RS3 regulated the systemic metabolic dysbiosis in obese mice and adjusted 26 abnormal metabolites in amino acids and lipids metabolism, many of which are related to the microbiome. More importantly, we found that the anti-obesity effect of Ce-RS3 can be transferred by fecal transplantation. The beneficial effects of Ce-RS3 might derive from gut microbiota changes, which might improve obesity and metabolic inflammation by altering host-microbiota interactions with impacts on the metabolome. In conclusion, Ce-RS3 can be used as a prebiotic with potential value for the treatment of obesity.

目的:探讨小儿珠珀散治疗感冒的药物效应,为临床应用提供参考.方法:针对感冒过程中儿童可能出现的发热、咳嗽、痰多、消化不良等症状,分别利用脂多糖(LPS)致热家兔,氨水诱发小鼠咳嗽,酚红排泌法、半固体营养糊小鼠肠推进实验探讨小儿珠珀散对症治疗的作用.结果:小儿珠珀散可降低小鼠耳肿胀程度、家兔体温上升幅度;延长小鼠咳嗽潜伏期,减少咳嗽次数,减少小鼠气管酚红排出量;促进小鼠胃排空,增加小鼠小肠推进率(P均＜0.05).结论:小儿珠珀散可缓解感冒过程中的多种症状.

目的:评价芭蕉芋RS3型抗性淀粉对肥胖型高脂血症小鼠的减肥降脂作用,并进行安全性评价.方法:KKAy小鼠高脂饲料喂养20周,建立肥胖型高脂血症模型,随机分为模型组,辛伐他汀组(4 mg·kg-1),RS3抗性淀粉高、低剂量组(2,1 g·kg-1),并以维持饲料喂养的小鼠为正常组.给药组分别灌胃相应药物,正常组及模型组给予等量去离子水.8周后,处死小鼠,检测血清中总胆固醇(TC),甘油三酯(TG),高密度脂蛋白胆固醇(HDL-C),低密度脂蛋白胆固醇(LDL-C)及天门冬氨酸氨基转移酶(AST),丙氨酸氨基转氨酶(ALT)的含量,精密称量脂肪质量,计算脂/体比、体脂率及Lee's指数,伊红-苏木素(HE)染色观察肝脏及脂肪组织的病理学变化;采用限量法评价RS3抗性淀粉的安全性,连续观察14d,记录小鼠毒副反应情况.结果:RS3抗性淀粉高剂量能显著降低小鼠体质量、脂肪质量、体脂率、脂/体比、Lee's指数,以及血清中TC,TG,LDL-C,AST,ALT水平(P＜0.05).组织形态学检测显示,给予RS3抗性淀粉可以明显改善肝脏组织的脂肪病变情况,保肝作用明显,可以抑制脂肪细胞膨大,降低小鼠脂肪累积,以高剂量为优;急毒实验小鼠按36 g·kg-1剂量给药后,动物未出现中毒反应及死亡.结论:RS3型芭蕉芋抗性淀粉具有良好的减肥降脂作用,且以高剂量组为佳,最大给药量证明无毒副作用,临床常用剂量安全可靠.

To characterize Salvia miltiorrhiza acetyl-CoA C-acetyltransferase (SmAACT), SmAACT was cloned, expressed in Escherichia coli and used for catalysis in vitro. The results showed that the SmAACT gene was 1212 bp in length, encoding a protein of 403 amino acids with a theoretical molecular mass of 41.20 kDa. Bioinformatic analysis showed that it had thiolase-related domains, including the C-terminal conserved domain, suggesting that it may be a member of the thiolase family. Catalytic reaction in vitro proved that the obtained SmAACT had the function of catalyzing the production of one molecule of acetoacetyl-CoA from two molecules of acetylCoA. Interestingly, we found that SmAACT had diverse wild genotypes which contained two main phylogenetic subclades, and those wild genotypes could cause non-synonymous mutations in proteins. Catalytic reaction of those wild mutants in vitro further proved that spontaneous mutations of SmAACT had great impact on their catalytic efficiencies. This study lay a good foundation for biosynthesis of tanshinones and molecular breeding of S. miltiorrhiza.

目的:观察孜亚比提片对2型糖尿病大鼠的降糖效果,并基于代谢组学技术分析其降糖作用机制.方法:根据临床调研结果对不同组分来源的孜亚比提片进行筛选,筛选出最佳组方配比;并以2型糖尿病大鼠为实验对象进行药效学实验研究,正常组和模型组分别给予相同剂量的生理盐水、孜亚比提片组给予300 mg·kg-1孜亚比提片、二甲双胍组给予300 mg·kg-1盐酸二甲双胍片,每组8只,记录给药后实验组的空腹血糖值及体质量变化并进行数据分析;同时利用超高效液相色谱-线性离子阱/静电场轨道阱组合式高分辨质谱联用(UHPLC-LTQ Orbitrap MS)技术对大鼠血清进行代谢组学分析,采用主成分分析(PCA),偏最小二乘判别分析(OPLS-DA)对不同组别大鼠血清代谢物进行分析,鉴定出潜在生物标记物,并分析维吾尔族药孜亚比提片的降糖作用机制.结果:与正常组比较,模型组大鼠的空腹血糖值升高(P＜0.01);与模型组比较,孜亚比提片组大鼠状态整体较好,毛色光滑,其空腹血糖值也有所下降(P＜0.05),表明孜亚比提片对2型糖尿病大鼠有一定的降糖作用;根据LC-MS代谢组学数据结果最终得到8个生物标记物.与模型组相比孜亚比提片组中L-valine,propionylcarnitine含量均降低(P＜0.05),sphingosine-1-phosphate,LPC(16∶1/0∶0),LPC(18∶0/0∶0),LPC(18∶2/0∶0),LPC(20∶ 1/0∶0),PC(19∶0/0∶ 0)的含量均升高(P＜0.05).结论:孜亚比提片能够降低2型糖尿病大鼠血糖值,并能改善2型糖尿病大鼠的一般生理特征,其作用机制可能与改善氨基酸代谢,脂质代谢有关.

Objective To analyze the variation types kaunene synthase-like (KSL) gene in Danshen (Salvia Root, Radix Salviae Miltiorrhizae) from different producing areas in China, and investigate primarily the influence of KSL gene polymorphism on enzyme-catalyzed reaction of its codases.Methods Specific primers were designed, and full-length KSL genes of Danshen from different producing areas (Henan, Shandong, Sichuan, Hubei, Shanxi, Neimonggu, Hebei) in China were homologously cloned.The variation types were compared and mutation samples were selected to establish a prokaryotic expression system and inducible expression in vitro.The enzyme-catalyzed products of KSL gene were detected by using gas chromatography-mass spectrometer (GS-MS) , and influence of KSL gene polymorphism of Danshen on protease catalytic efficiency was further analyzed.Results The full-length KSL gene was 1 788 bp, and there were totally 595 amino acid residues encoded.The sequencing results showed that there were polymorphisms in KSL gene of Danshen from different producing areas in China, which could cause synonymous and non-synonymous mutations.The catalytic efficiency of different genotypes of KSL was significantly different, which was the highest in Shanxi and the lowest in Inner Mongolia.The KSL gene containing A-type variation mutated at 1 047 bp in Danshen from4 producing areas including Shanxi, Hubei, Shandong, and Sichuan, and the catalytic efficiency of which was significantly higher than those containing no Atype variation in Danshen from4 producing areas including Henan, Hebei and Inner Mongolia.Conclusion KSL genes have gene polymorphisms indeed in Danshen from different producing areas.The non-synonymous mutations induced by polymorphisms can further lead to changes of amino acid sequences and finally resulting in difference in the catalytic efficiency of encoded enzymes.The polymorphisms of key enzyme genes in the synthesis pathway of tanshinones can be taken as a starting point to provide further reference for revealing the accumulation mechanism of tanshinones and breeding the excellent germplasm of Danshen.

目的:观察桑叶含药血清对脂肪细胞株3T3-L1胰岛素抵抗(IR)模型葡萄糖消耗量及细胞活力的影响,筛选出桑叶含药血清的最佳浓度,并检测其对炎症因子含量的影响,探讨可能的作用机制.方法:取对数生长期3T3-L1前脂肪细胞,10 mg·L-1胰岛素(Ins),0.25 mmol· L-1地塞米松(DEX),0.5 mmol·L-13-异丁基-1-甲基黄嘌呤(IBMX)诱导48 h后,10 mg·L-1 Ins再次诱导48 h,待其分化为成熟脂肪细胞后,1μmol·L-DEX诱导96 h以建立IR细胞模型.桑叶水提物含药血清培养12,24,36,72 h后,采用葡萄糖氧化酶法检测桑叶含药血清培养后细胞葡萄糖的消耗量,采用噻唑蓝(MTT)比色法检测桑叶含药血清培养后IR模型的细胞活力,酶联免疫吸附测定法(ELISA)检测桑叶含药血清对细胞肿瘤坏死因子-α(TNF-α)含量的影响,蛋白免疫印迹法(Western blot)测定桑叶含药血清对胰岛素信号通路胰岛素受体(InsR),胰岛素受体底物(IRS),磷酸化IRS1(p-IRS1),葡萄糖转运蛋白4(GLUT4)蛋白表达的影响.结果:桑叶含药血清可显著提高IR细胞葡萄糖消耗量(P＜0.01),增强细胞活力(P＜0.01),降低炎症因子TNF-α的含量(P＜0.01),调节胰岛素信号通路下游蛋白的表达量(P＜0.05,P＜0.01).结论:桑叶可明显改善3T3-L1细胞IR状态,其作用机制可能与抑制TNF-α表达、促进胰岛素信号通路蛋白的表达有关.

BACKGROUND:The causal link between tobacco smoke exposure (TSE) and numerous severe respiratory system diseases (RSD), including chronic bronchitis, chronic obstructive pulmonary disease, and lung cancer, is well established. However, the pathogenesis of TSE-induced RSD remains incompletely understood. This research aims to detect the pathogenetic mechanisms and potential therapeutic targets of TSE-induced RSD.METHODS:This study employed TSE model which rats were exposed to a concentration of 60% tobacco smoke in a toxicant exposure system for four weeks. Tandem mass tags (TMT) labeled quantitative proteomics combined with off-line high pH reversed-phase fractionation, and nano-liquid chromatography-mass spectrometry method (off-line high pH RPF-nano-LC-MS/MS) were adopted to detect differentially expressed proteins (DEPs) in the lung tissues of the TSE model rats and to compare them with those in control. The accuracy of the results was verified by western blot.RESULTS:Compared with the control group, 33 proteins in the TSE model group's lung tissues showed significant differential expression. Analysis based on the Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways indicated that, several biological pathways, such as the steroid biosynthesis pathway, were involved and played significant roles in the pathogenesis of the experimental group's TSE.CONCLUSIONS:These findings make a crucial contribution to the search for a comprehensive understanding of TSE-induced RSD's pathogenesis, and furthermore provide guidance for the diagnosis and treatment of TSE-induced RSD.

Objective:To establish a quick identification method of UPLC-LTQ Orbitrap MS ( ultra-high performance liquid chromatography-linear trap/orbitrap high resolution mass spectrometry) for the identification of chemical constit-uents in Danshen( Radix et Rhizoma Salviae Miltiorrhizae) . Methods:The negative and positive ion model was used for Orbitrap MS scanning, the components were identified by high resolution data and the secondary mass spectrum informa-tion. Results:Debris features and mass spectrum cracking rule were concluded in the standard substance of tanshinone IIA and Dan phenolic acid B. 70 components including diterpenes, phenolic acids, and quinones were discovered from extract of Radix et Rhizoma Salviae Miltiorrhizae. Conclusion: This method could provide rapid qualitative analysis for the chemical components of Radix et Rhizoma Salviae Miltiorrhizae, thus to provide technical support for the quality con-trol of it and its pharmacodynamics research. It can also provide reference for comprehensive characterization of complex chemical constituents of Chinese herbs.

With increasing air pollution, chronic bronchitis (CB) has become a major public health problem worldwide. Previous studies have shown beneficial effects of Bergenin (Ber) on chronic bronchitis. To facilitate understanding of the pathogenesis underlying CB as well as to elucidate the Ber therapeutic mechanism, it is crucial to confirm the rational biomarkers of CB and its treatment. This study aimed to investigate the preventive chronic bronchitis mechanism of Ber by applying a serum metabolomics strategy. In this study, 18 Sprague-Dawley rats were randomly divided into three groups, with six rats in each group. Rats from the CB and Ber groups were exposed to tobacco smoke for 1 hd-1 (1 h per day) for 28 days. Ber was administered orally to Ber rats 3 h after exposure every day, and the others were administered water. According to the morphometric analysis of the airway epithelium and the count of white blood cells in the bronchoalveolar lavage fluid, Ber suppressed the infiltration of inflammatory cells, inhibited the secretion of mucus, and reduced white blood cells in bronchoalveolar lavage fluid. The metabolic profiles of sera were analyzed by multivariate statistical analyses, including PCA, PLS-DA and OPLS-DA models, and revealed that the levels of thirteen metabolites were significantly changed and identified as potential biomarkers in the CB group and Ber group. The results suggested that the therapeutic mechanism of Ber may be related to the regulation of dysfunctions in glycerophospholipid, tryptophan, arginine and proline metabolism induced by CB, and changes in arachidonic acid metabolism.

目的:观察桑叶降糖作用,分析桑叶对2型糖尿病(T2DM)大鼠肝脏Toll样受体及其下游信号元件基因表达的影响,初步探讨其降糖机制.方法:选取SPF级雄性Wistar大鼠46只,高脂饲料饲喂8周后,腹腔注射链脲佐菌素(STZ),以制备T2DM模型.根据血糖值将模型大鼠随机分成模型组、阳性药二甲双胍(0.2 g/kg)组和桑叶高、低剂量(3 g/kg,1 g/kg)组,每组12只,另取10只大鼠为正常对照组.灌胃给药,持续12周,每周测其空腹血糖,并于第11周时进行OGTT和ITT实验.以上实验结束后免疫吸附试验(ELISA)试剂盒测定血清中胰岛素、IL-6、肿瘤坏死因子-(TNF-)和C反应蛋白(CRP)水平,实时PCR法测定大鼠肝脏TLRs及MyD88、TRAF-6、核因子-B mRNA水平.结果:大鼠给药桑叶后,血糖值降低,饮水量明显减少,差异有统计学意义(P<0.05),糖耐量得到显著改善,血清中IL-6、CRP水平明显降低,差异有统计学意义(P<0.05),TLR7、TLR8、TLR9以及其下游信号元件TRAF-6、MyD88、核因子-B mRNA相对表达均较模型组明显降低,差异有统计学意义(P<0.05).结论:桑叶降糖可能与其抑制肝脏组织TLRs及其下游信号表达,降低炎性反应有关.

Objective To analyze the content of active components in fresh Danshen Radix Salviae Miltiorrhizae,Red Sage Root) from different producing areas,and review reasonably the medicinal quality of fresh Danshen.Methods The content of 5 salvianolic acids and 12 tanshinones in fresh Danshen,from 7 different producing areas,were determined by applying LTQ-Orbitrap XL high-resolution mass spectrometer.On this basis,all content were analyzed by using principal component analysis (PCA) and partial least square (PLS) analysis.Results The content of active components in fresh Danshen from the same producing area were basically the same,and difference was significant in content of active components among fresh Danshen from different producing areas.The content of salvianolic acids was the highest in fresh Danshen from Hubei,and content of tanshinone Ⅰ and tanshinone ⅡA were the highest in fresh Danshen from Sichuan and Henan.The analysis results of PCA and PLS showed that the distribution of most active components had a distinct regional specificity.Conclusion The difference is significant in quality of fresh Danshen from different producing areas,and they have a distinct regional specificity.Using fresh Danshen as quality evaluation materials,that is a useful supplement to review quality of raw medicinal materials based on dry herbs.It explored a new and more comprehensive approach for reviewing other Chinese medicinal materials.