Different metabolic profiles of K1 serotype and non-serotype K1 and K2 Klebsiella pneumoniae isolates in oral infection mice model

K1 or K2 serotype Klebsiella pneumoniae isolate caused clinical pyogenic liver abscess (KLA) infection is prevalent in many areas. It has been identified that K1 or K2 serotype K. pneumoniae isolates caused KLA infection in mice by oral inoculation. In our study, K1 serotype K. pneumoniae isolate Kp1002 with hypermucoviscosity (HV)-positive phenotype caused KLA infection in C57BL/6 mice by oral inoculation. Simultaneously, non-serotype K1 and K2 isolate Kp1014 with HV-negative phenotype failed to cause KLA infection in the same manner. It seems that gastrointestinal tract translocation is the pathway by which K1 or K2 serotype K. pneumoniae caused KLA infection. Liquid chromatography-tandem mass spectrometry was used to further analyze metabolic profile changes in mice with KLA infection. Data showed that after Kp1002 or Kp1014 oral inoculation, serum Phosphatidylcholine (PC) and Lysophosphatidylcholine (LPC) levels significantly changed in mice. Some PC and LPC molecules showed changes both in the Kp1002 KLA group and the Kp1014 no-KLA group compared with the control group. The level of 18:1/18:2-PC significantly changed in the Kp1002 KLA group compared with the control group, but showed no change between the Kp1014 no-KLA group and the control group. The level of 18:1/18:2-PC might have been particularly affected by KLA infection caused by K1 serotype K. pneumoniae Kp1002. It may be a potential biomarker for KLA infection.     

耐药口腔鳞癌Tca8113胞外代谢产物的代谢组学分析

目的 探讨口腔鳞癌Tca8113细胞在产生耐药性后胞外代谢产物的变化,寻找差异代谢物.方法 制备胞外代谢产物样本,运用氢谱核磁共振(1H-NMR)获取口腔鳞癌Tca8113细胞和耐药Tca8113/CBP细胞的胞外代谢物图谱,偏最小二乘判别分析(PLS-DA)得出对区分2组细胞贡献较大的差异性变量,将同时满足VIP＞ 1.0和单维统计P＜0.05的代谢物确定为最终的差异代谢物.结果 基于氢谱核磁共振的代谢组学方法可以区分Tca8113和Tca8113/CBP;最终的差异性代谢物有醋酸盐、牛磺酸、丝氨酸、葡萄糖和亮氨酸,它们涉及到了蛋白质代谢、糖代谢和三羧酸循环.结论 应用代谢组学方法成功找到了耐药Tca8113细胞的差异代谢物.     

基于指纹图谱及化学模式识别研究盐制对五子衍宗丸化学成分的影响

目的建立不同炮制品组方五子衍宗丸(Wuzi Yanzong Pills,WYP)的HPLC指纹图谱,研究盐制对WYP化学成分的影响。方法采用HPLC法建立不同炮制品组方WYP的指纹图谱,并进行相似度评价,采用方差分析、聚类分析(CA)、主成分分析(PCA)与偏最小二乘法-判别分析(PLS-DA)对结果进行评价。结果 WYP全生品组、药典组、盐制组指纹图谱相似度均大于0.9,标定了18个共有峰,利用对照品指认出绿原酸(5号峰)、鞣花酸(10号峰)、金丝桃苷(11号峰)、异槲皮苷(12号峰)、毛蕊花糖苷(13号峰)、紫云英苷(14号峰)、槲皮素(15号峰)、山柰酚(17号峰)、五味子醇甲(18号峰)9个成分,归属性分析表明峰2来源于枸杞子,峰3、5、7、9、11、12、14、15、17来源于菟丝子,峰6、8、10、16来源于覆盆子,峰13来源于车前子,峰18来源于五味子,峰1是覆盆子和五味子共有成分,峰4是枸杞子和覆盆子共有成分。药物炮制后组方WYP未出现色谱峰的增加,但峰面积发生变化。通过CA将WYP全生品组、药典组、盐制组聚为3类,PCA筛选出4个主成分,PLS-DA标记出峰1、13(毛蕊花糖苷)、10(鞣花酸)、7、4、11(金丝桃苷)、12(异槲皮苷)、3、5(绿原酸)共9个差异性成分;绿原酸、鞣花酸、金丝桃苷、异槲皮苷、山柰酚、五味子醇甲及峰1、3、4、7、8共11个成分可能是不同炮制品组方WYP的差异标志物。结论建立的指纹图谱测定方法稳定、可靠,炮制后组方WYP的化学成分发生变化,结果可为WYP中如何选用药物炮制品提供一定的依据。     

Alpha-naphthylisothiocyanate modulates hepatobiliary transporters in sandwich-cultured rat hepatocytes

Alpha-naphthylisothiocyanate (ANIT) induces intra-hepatic cholestasis mixed with hepatocellular injury mainly by bile ductular damage. However, its direct effect on hepatic parenchymal cells (hepatocytes) is unclear. Sandwich-cultured rat hepatocytes (SCRH) were applied to clarify this question. Though cytotoxicity was not observed (0–180 μM) in ANIT-treated SCRH, metabonomics analysis of the hepatocytes revealed a shift in the metabolic pattern and a decrease in cellular cholesterol level, accompanied by an increase in total bile acids after 48 h ANIT (5–45 μM) treatment. To assess the function of major hepatic bile acid transporters, the accumulation and efflux of [D-Pen^2,5]-enkephalin (DPDPE), 5 (and 6)-carboxy-2′,7′-dichlorofluorescein (CDF) diacetate promoiety and deuterium-labeled sodium taurocholate (d8-TCA) were measured. ANIT incubation for either 30 min or 48 h led to dose-dependent decreases in the biliary excretion index (BEI) of DPDPE and CDF, as well as the intracellular accumulation of d8-TCA, CDF and DPDPE. The basolateral efflux of d8-TCA was also decreased with its BEI barely changed. mRNA expression of multiple uptake transporters and bile acid synthesizing enzymes was down-regulated after 48 h incubation. In conclusion, ANIT could directly induce retention of bile acids in hepatocytes by inhibiting the function of bile acid transporters, which might contribute to its cholestatic effect.     

Non-targeted metabolomic biomarkers and metabotypes of type 2 diabetes: A cross-sectional study of PREDIMED trial participants

Aim

To characterize the urinary metabolomic fingerprint and multi-metabolite signature associated with type 2 diabetes (T2D), and to classify the population into metabotypes related to T2D.

Metabolic profiling studies on the toxicological effects of realgar in rats by H NMR spectroscopy

The toxicological effects of realgar after intragastrical administration (1 g/kg body weight) were investigated over a 21 day period in male Wistar rats using metabonomic analysis of ¹H NMR spectra of urine, serum and liver tissue aqueous extracts. Liver and kidney histopathology examination and serum clinical chemistry analyses were also performed. ¹H NMR spectra and pattern recognition analyses from realgar treated animals showed increased excretion of urinary Kreb's cycle intermediates, increased levels of ketone bodies in urine and serum, and decreased levels of hepatic glucose and glycogen, as well as hypoglycemia and hyperlipoidemia, suggesting the perturbation of energy metabolism. Elevated levels of choline containing metabolites and betaine in serum and liver tissue aqueous extracts and increased serum creatine indicated altered transmethylation. Decreased urinary levels of trimethylamine-N-oxide, phenylacetylglycine and hippurate suggested the effects on the gut microflora environment by realgar. Signs of impairment of amino acid metabolism were supported by increased hepatic glutamate levels, increased methionine and decreased alanine levels in serum, and hypertaurinuria. The observed increase in glutathione in liver tissue aqueous extracts could be a biomarker of realgar induced oxidative injury. Serum clinical chemistry analyses showed increased levels of lactate dehydrogenase, aspartate aminotransferase, and alkaline phosphatase as well as increased levels of blood urea nitrogen and creatinine, indicating slight liver and kidney injury. The time-dependent biochemical variations induced by realgar were achieved using pattern recognition methods. This work illustrated the high reliability of NMR-based metabonomic approach on the study of the biochemical effects induced by traditional Chinese medicine.     

不同产地麦冬1H-NMR模式识别研究

目的 建立一种基于氢核磁共振-模式识别的不同产地麦冬鉴别新方法。方法 以¹H-NMR技术测定样品的全成分信息,并转化成数据矩阵,采用模式识别法中的主成分分析(PCA)、偏最小二乘法-判别分析(PLS-DA)以及聚类分析(HCA)进行识别分析。结果 氢核磁共振-模式识别法能有效地鉴别不同产地的麦冬样本。结论 氢核磁共振-模式识别法是一种有效的药材分类鉴别方法,可作为药材质量控制的手段之一。     

PLS-DA结合熵权TOPSIS法综合评价不同产地苍耳子

目的 综合评价不同产地苍耳子的质量。方法 采用HPLC法测定不同产地苍耳子中羧基苍术苷、苍术苷、新绿原酸、绿原酸、隐绿原酸、噻嗪双酮苷、1,5-二咖啡酰奎宁酸、3,4-二咖啡酰奎宁酸、3,5-二咖啡酰奎宁酸和4,5-二咖啡酰奎宁酸,采用偏最小二乘判别分析（PLS-DA）和熵权优劣解距离法（EW-TOPSIS）法综合分析测定结果,寻找引起苍耳子产品质量的主要差异性物质,评价不同产地苍耳子质量的优劣。结果 10种成分在各自范围内线性关系良好;绿原酸、1,5-二咖啡酰奎宁酸、3,5-二咖啡酰奎宁酸、羧基苍术苷和新绿原酸是影响苍耳子产品质量的主要潜在标志物;EW-TOPSIS法分析结果显示江苏和山东产的苍耳子质量优。结论 建立的HPLC法操作便捷、结果准确,结合PLS-DA、EW-TOPSIS法可综合评价不同产地苍耳子的质量。     

细胞因子对脾虚水湿不化大鼠模型影响的曲线拟合分析

目的：探讨细胞因子对脾虚水湿不化大鼠模型综合影响的数学模型问题。方法：采用“饮食不节+久居水湿”的方法建立脾虚水湿不化大鼠模型,以PLS-DA法建立模型综合评价指标并计算主元得分,并对细胞因子与主元得分进行Linear、Quadratic及Cubic曲线拟合。结果：对11个大鼠脾虚水湿不化描述的指标进行PLS-DA综合评价,共提取一个有效主元成分,该主元得分与IL-4、IL-2和IFN-γ均具有良好的相关关系（r>0.5,P<0.05）;对IL-2、IL-4和IFN-γ与主元得分进行Linear、Quadratic及Cubic曲线拟合,结果发现,3种拟合方程中,IL-2、IFN-γ拟合方程均具有统计学意义（P<0.05）,但R2均小于0.4,这说明方程对结果的解释效果一般;但IL-4的Quadratic及Cubic曲线拟合不仅方程具有统计学意义（P<0.05）,且R²>0.6,方程对结果的解释效果好。结论：IL-4对脾虚水湿不化的影响并不呈线性,而符合Quadratic或Cubic方程,IL-4在脾虚水湿不化的发生过程中具有重要作用。     

A review of near infrared spectroscopy and chemometrics in pharmaceutical technologies

Near-infrared spectroscopy (NIRS) is a fast and non-destructive analytical method. Associated with chemometrics, it becomes a powerful tool for the pharmaceutical industry. Indeed, NIRS is suitable for analysis of solid, liquid and biotechnological pharmaceutical forms. Moreover, NIRS can be implemented during pharmaceutical development, in production for process monitoring or in quality control laboratories.This review focuses on chemometric techniques and pharmaceutical NIRS applications. The following topics are covered: qualitative analyses, quantitative methods and on-line applications. Theoretical and practical aspects are described with pharmaceutical examples of NIRS applications.