基于HPLC-Q-TOF-MS/MS技术的七味通痹口服液化学成分分析

目的采用高效液相色谱串联四极杆飞行时间质谱联用技术(HPLC-Q-TOF-MS/MS)对七味通痹口服液中的化学成分进行定性表征。方法色谱柱为Agilent ZORBAX SB-C_(18)(250 mm×4.6 mm,5μm),以甲醇-0.1%甲酸水溶液为流动相,梯度洗脱,体积流量1.0 mL/min,柱温25℃;采用电喷雾离子源,正、负离子全扫描模式下采集数据,根据精确质荷比、二级碎片离子信息,结合对照品和文献数据鉴定七味通痹口服液中的化学成分。结果共指认出七味通痹口服液制剂中72个化学成分,包括生物碱类、黄酮类、氨基酸类、酚酸类及苯丙素类等,其中18个化合物经对照品比对。结论该研究为七味通痹口服液的药效物质基础及质量控制的深入研究奠定了基础。     

山楂的化学成分、药理作用及质量标志物（Q-Marker）预测

山楂是我国传统的药材,具有健胃消食、行气散瘀、化浊调脂的功效。其主要化学成分类型为黄酮类、萜类、有机酸类、多糖类等。现代药理研究表明,山楂具有促进胃肠蠕动及软化心脑血管方面的作用。此外,还具有调节免疫、抗氧化、抗肿瘤、抑菌等作用。因山楂具有多基原、多功效、多炮制品的特点,而《中国药典》中指标成分仅为枸橼酸,故对山楂的化学成分、药理作用进行了总结概括,并根据质量标志物(Q-Marker)的概念从植物化学成分专有性、化学成分与有效成分、化学成分可测性等方面进行了分析,有机酸类和黄酮类中的槲皮素、金丝桃苷、芦丁、山柰酚、牡荆素-4′-鼠李糖苷、牡荆素、牡荆素鼠李糖苷可作为Q-Marker成分,为山楂的质量评价研究提供依据。     

黄连地黄汤辅助治疗肾阴虚损型糖尿病临床研究

目的:探讨黄连地黄汤联合沙格列汀治疗肾阴虚损型糖尿病疗效及对患者生活质量及血清摄食抑制因子(Nesfatin-1)、游离脂肪酸(FFA)水平的影响。方法:选取肾阴虚损型糖尿病患者106例,随机数字表法分为研究组与对照组,各53例。常规干预基础上对照组予以沙格列汀治疗,研究组在对照组基础上予以黄连地黄汤治疗。统计两组治疗前、后血糖、糖化血红蛋白(HbA1c)、空腹血糖(FPG)、餐后2 h血糖(2hPG)、血清Nesfatin-1、游离脂肪酸(FFA)含量、胰岛β细胞功能指数(HOMA-β)、空腹胰岛素(FINS)、胰岛素抵抗指数(HOMA-IR)水平、生活质量(DQOL)分值、不良反应。结果:研究组治疗后HbA1c、FPG、2hPG低于对照组(P<0.05); 治疗后研究组血清FFA水平低于对照组,血清Nesfatin-1水平高于对照组(P<0.05); 治疗后研究组HOMA-IR低于对照组,FINS、HOMA-β高于对照组(P<0.05); 治疗后研究组DQOL分值低于对照组(P<0.05); 研究组不良反应发生率(13.21%)与对照组(9.43%)比较无统计学差异(P>0.05)。结论:肾阴虚损型糖尿病患者采用沙格列汀联合黄连地黄汤治疗,能有效改善患者血糖、胰岛β细胞功能及血清Nesfatin-1、FFA水平,提升生活质量,且具有安全性。     

Amino acid signatures in the developing mouse retina

This study characterizes the developmental patterns of seven key amino acids: glutamate, γ-amino-butyric acid (GABA), glycine, glutamine, aspartate, alanine and taurine in the mouse retina. We analyze amino acids in specific bipolar, amacrine and ganglion cell sub-populations (i.e. GABAergic vs. glycinergic amacrine cells) and anatomically distinct regions of photoreceptors and Müller cells (i.e. cell bodies vs. endfeet) by extracting data from previously described pattern recognition analysis. Pattern recognition statistically classifies all cells in the retina based on their neurochemical profile and surpasses the previous limitations of anatomical and morphological identification of cells in the immature retina. We found that the GABA and glycine cellular content reached adult-like levels in most neurons before glutamate. The metabolic amino acids glutamine, aspartate and alanine also reached maturity in most retinal cells before eye opening. When the overall amino acid profiles were considered for each cell group, ganglion cells and GABAergic amacrine cells matured first, followed by glycinergic amacrine cells and finally bipolar cells. Photoreceptor cell bodies reached adult-like amino acid profiles at P7 whilst Müller cells acquired typical amino acid profiles in their cell bodies at P7 and in their endfeet by P14. We further compared the amino acid profiles of the C57Bl/6J mouse with the transgenic X-inactivation mouse carrying the lacZ gene on the X chromosome and validated this animal model for the study of normal retinal development. This study provides valuable insight into normal retinal neurochemical maturation and metabolism and benchmark amino acid values for comparison with retinal disease, particularly those which occur during development.     

Postprandial coagulation activation in overweight individuals after weight loss: Acute and long-term effects of a high-monounsaturated fat diet and a low-fat diet

Diet is important in the prevention of cardiovascular disease, and it has been suggested that a high-MUFA diet is more cardioprotective than a low-fat diet. We hypothesised that the postprandial thrombotic risk profile is improved most favourably by a high-MUFA diet compared with a low-fat diet. This was tested in a parallel intervention trial on overweight individuals (aged 28.4 (SD 4.7) years) randomly assigned to a MUFA-diet (35-45% of energy as fat; > 20% as MUFA, n = 21) or a low-fat (LF) diet (20-30% of energy as fat, n = 22) for 6 months after a weight loss of ~ 10%. All foods were provided free of charge from a purpose-built supermarket. Meal tests designed after the same principles were performed before and after the dietary intervention, and blood samples were collected at 8.00 h (fasting), 12.00 h, and 18.00 h and analysed for factor VII coagulant activity (FVII:C), activated FVII, fibrinogen, prothrombin fragment 1 + 2 (F1 + 2), D-dimer, plasminogen activator inhibitor (PAI:Ag), and thrombin activatable fibrinolysis inhibitor. There were significant postprandial increases in F1 + 2 and D-dimer before and after dietary intervention, with significantly lower values after 6 months. No significant differences were observed between the postprandial changes induced by the two diets. The postprandial decrease in FVII:C and PAI:Ag did not differ before and after intervention, irrespective of the diets. Our findings suggest postprandial coagulation activation in overweight subjects with more pronounced acute than long-term effects. We observed similar effects of the MUFA diet and the LF diet on the postprandial prothrombotic risk profile.     

The effect of docosahexaenoic acid on bone microstructure in young mice and bone fracture in neonates

Background

As low bone mineral density is a risk factor for fracture in childhood, optimizing age appropriate bone mass is recommended and might lower the impact of bone loss related to age. Consumption of omega-3 polyunsaturated fatty acids (PUFAs), including eicosapentaenoic and docosahexaenoic (DHA) acids have been shown to beneficially modulate bone metabolism. The objective of this study was to determine the incidence of fracture in neonates receiving a fish compared with soybean oil–based intravenous lipid emulsion and evaluate the effect of varying dietary omega-3 PUFA consumption on growing bone in young mice.

Materials and methods

Eligibility criteria for the clinical study included gestational age ≤37 wk and parenteral nutrition–dependence for ≥4 wk. Radiographs were reviewed after lipid initiation to identify radiologic bone fracture. The animal study evaluated female C57/Bl6 mice randomized into one of five groups from age 3–12 wk, at which time femurs were harvested for micro–computed tomography and light microscopy analysis.

Results

A lower incidence of bone fracture was found in neonates maintained on fish compared with soybean oil. In the animal study, findings suggest the DHA diet provides the best protection against trabecular bone loss as evidenced by increased bone volume fraction, increased trabecular number, and decreased trabecular separation on micro–computed tomography. These protective effects appeared to affect the bone microstructure alone.

Conclusions

The lower fracture risk observed in fish oil fed neonates in combination with the protective effects of DHA observed in the femurs of young C57/BL6 mice suggest an important role for omega-3 PUFAs on bone growth.     

Effects of quercetin and fish n‐3 fatty acids on testicular injury induced by ethanol in rats

The aim of this study was to investigate the effects of quercetin and fish n‐3 fatty acids on the changes in testis induced by ethanol. Forty‐five rats divided into five groups, control, ethanol, ethanol+quercetin, ethanol+fish n‐3 fatty acids and ethanol+quercetin+fish n‐3 fatty acids. At the end of 8 weeks, all the rats were sacrificed. Degenerative changes in histopathological analyses, the decreased body weight gain and seminiferous tubule diameters in ethanol group have been observed. TUNEL assay also showed an increase in apoptotic cell number. The activities of superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GSH‐Px), xanthine oxidase (XO) and testosterone levels were decreased as well as the levels of malondialdehyde (MDA) and nitric oxide (NO) were increased in ethanol group. Histopathological changes caused by ethanol have been improved by quercetin and fish n‐3 fatty acids. It was also found that protection was provided by increasing SOD, CAT and GSH‐Px activities in groups administered quercetin, fish n‐3 fatty acids and quercetin+fish n‐3 fatty acids, and by decreasing the levels of MDA and NO in groups administered both quercetin and fish n‐3 fatty acids together. These results suggest that quercetin and fish n‐3 fatty acids are beneficial agents to reduce testicular injury induced by ethanol except for testosterone levels.     

Syntrophic exchange in synthetic microbial communities

Metabolic crossfeeding is an important process that can broadly shape microbial communities. However, little is known about specific crossfeeding principles that drive the formation and maintenance of individuals within a mixed population. Here, we devised a series of synthetic syntrophic communities to probe the complex interactions underlying metabolic exchange of amino acids. We experimentally analyzed multimember, multidimensional communities of Escherichia coli of increasing sophistication to assess the outcomes of synergistic crossfeeding. We find that biosynthetically costly amino acids including methionine, lysine, isoleucine, arginine, and aromatics, tend to promote stronger cooperative interactions than amino acids that are cheaper to produce. Furthermore, cells that share common intermediates along branching pathways yielded more synergistic growth, but exhibited many instances of both positive and negative epistasis when these interactions scaled to higher dimensions. In more complex communities, we find certain members exhibiting keystone species-like behavior that drastically impact the community dynamics. Based on comparative genomic analysis of >6,000 sequenced bacteria from diverse environments, we present evidence suggesting that amino acid biosynthesis has been broadly optimized to reduce individual metabolic burden in favor of enhanced crossfeeding to support synergistic growth across the biosphere. These results improve our basic understanding of microbial syntrophy while also highlighting the utility and limitations of current modeling approaches to describe the dynamic complexities underlying microbial ecosystems. This work sets the foundation for future endeavors to resolve key questions in microbial ecology and evolution, and presents a platform to develop better and more robust engineered synthetic communities for industrial biotechnology.Microbes are abundantly found in almost every part of the world, living in communities that are diverse in many facets. Although it is clear that cooperation and competition within microbial communities is central to their stability, maintenance, and longevity, there is limited knowledge about the general principles guiding the formation of these intricate systems. Understanding the underlying governing principles that shape a microbial community is key for microbial ecology but is also crucial for engineering synthetic microbiomes for various biotechnological applications (1–3). Numerous such examples have been recently described including the bioconversion of unprocessed cellulolytic feedstocks into biofuel isobutanol using fungal–bacterial communities (4) and biofuel precursor methyl halides using yeast–bacterial cocultures (5). Other emerging applications in biosensing and bioremediation against environmental toxins such as arsenic (6) and pathogens such as Pseudomonas aeruginosa and Vibrio cholerae have been demonstrated using engineered quorum-sensing Escherichia coli (7, 8). These advances paint an exciting future for the development of sophisticated multispecies microbial communities to address pressing challenges and the crucial need to understand the basic principles that enables their design and engineering.An important process that governs the growth and composition of microbial ecosystems is the exchange of essential metabolites, known as metabolic crossfeeding. Entomological studies have elucidated on a case-by-case basis the importance of amino acids in natural interkingdom and interspecies exchange networks (9–11). Recent comparative analyses of microbial genomes suggest that a significant proportion of all bacteria lack essential pathways for amino acid biosynthesis (2). These auxotrophic microbes thus require extracellular sources of amino acids for survival. Understanding amino acid exchange therefore presents an opportunity to gain new insights into basic principles in metabolic crossfeeding. Recently, several studies have used model systems of Saccharomyces cerevisiae (12), Saccharomyces enterica (13), and E. coli (14–16) to study syntrophic growth of amino acid auxotrophs in coculture environments. Numerous quantitative models have also been developed to describe the behavior of these multispecies systems, including those that integrate dynamics (17, 18), metabolism (19–21), and spatial coordination (22). Although these efforts have led to an improved understanding of the dynamics of syntrophic pairs and the energetic and benefits of cooperativity in these simple systems (23), larger more complex syntrophic systems have yet to be explored.Here, we use engineered E. coli mutants to study syntrophic crossfeeding, scaling to higher-dimensional synthetic ecosystems of increasing sophistication. We first devised pairwise syntrophic communities that show essential and interesting dynamics that can be predicted by simple kinetic models. We then increased the complexity of the interaction in three-member synthetic consortia involving crossfeeding of multiple metabolites. To further increase the complexity of our system, we devised a 14-member community to understand key drivers of population dynamics over short and evolutionary timescales. Finally, we provide evidence for widespread trends of metabolic crossfeeding based on comparative genomic analysis of amino acid biosynthesis across thousands of sequenced genomes. Our large-scale and systematic efforts represent an important foray into forward and reverse engineering synthetic microbial communities to gain key governing principles of microbial ecology and systems microbiology.     

妊娠期肝内胆汁淤积症胆汁酸、肝酶指标对妊娠结局的影响

目的：探讨妊娠期肝内胆汁淤积症患者的胆汁酸、肝酶指标对患者妊娠结局的影响,为临床诊治妊娠期肝内胆汁淤积症患者给予参考。方法选择2012年1月～2013年10月我院收治的170例妊娠期肝内胆汁淤积症患者作为临床研究对象,对所有患者均给予胆汁酸、肝酶指标的检测,根据检测结果将患者分为轻度、中度和重度,并以同期我院收治的150例正常健康孕妇作为对照组,观察比较四组孕妇的妊娠结局。结果产后出血方面：重度妊娠期肝内胆汁淤积症患者的发生率最高,差异有统计学意义（P＜0.05）;重度妊娠期肝内胆汁淤积症患者剖宫产比例更高,新生儿窒息率更高,低体重儿比例高,新生儿Apgar评分低,与其他组别对比差异有统计学意义。结论对妊娠期肝内胆汁淤积症检测胆汁酸、肝酶指标,有助于判断患者的疾病严重程度,进而评估产妇和新生儿结局。