Identifying muscle hemorrhage in rat cadavers with advanced decomposition by FT-IR microspectroscopy combined with chemometrics

The identification of muscle hemorrhage in a cadaver that is in an advanced stage of decomposition is an important but challenging task. Our study investigated whether Fourier transform infrared (FT-IR) microspectroscopy in conjunction with chemometrics could identify muscle hemorrhage using rat cadavers with advanced decomposition. In this study, an intramuscular blood injection method, instead of a mechanical injury method, was used to construct a muscle hemorrhage model, and the modeling idea of muscle hemorrhage identification was to discriminate and classify hemoglobin-leaking myofibrils from negative myofibrils. First, the optical images of hematoxylin/eosin (H&E) stained hemorrhagic muscle at different postmortem intervals (PMIs) were observed and showed that the morphological features of whole erythrocytes disappeared since the PMI of 4 d. Subsequently, principle component analysis (PCA) was performed and indicated that the biochemical differences in protein structures between fresh erythrocytes and myofibrils can be detected by the IR spectroscopic method. Ultimately, several classification models based on the partial least square discriminant analysis (PLS-DA) algorithm were successfully constructed for different PMIs and PMI ranges and achieved great prediction performances in external validations. This preliminary study demonstrates the feasibility of using FT-IR microspectroscopy combined with chemometrics as a potential approach for identifying muscle hemorrhage in cadavers with advanced decomposition for offering vital evidences in judicial process.     

基于UPLC-Q/TOF-MS分析江西特色炮制工艺对枳壳化学成分的影响

目的:比较不同特色炮制工艺对枳壳化学成分的影响,筛选优势饮片品种。方法:采用UPLC-Q/TOF-MS检测枳壳中化学成分,选择ACQUITY UPLC BEH C18色谱柱(2. 1 mm×100 mm,1. 7μm),流动相0. 1%甲酸水溶液(A)-0. 1%甲酸乙腈溶液(B)梯度洗脱(0～10 min,5%～35%B;10～18 min,35%～75%B;18～21 min,75%～100%B;21～24 min,100%B;24～24. 1 min,100%～5%B;24. 1～28 min,5%B),电喷雾离子源(ESI),正离子模式,质谱扫描范围m/z 50～1 200,通过对照品、相对分子质量、质谱裂解规律和文献信息鉴定枳壳甲醇提取液的化学成分。利用SIMCA-P 13. 0软件建立枳壳各炮制品主成分分析(PCA)模型和偏最小二乘法-判别分析(PLS-DA)模型,获取PCA得分图,PLS-DA载荷图和变量重要性投影(VIP)值,筛选造成枳壳炮制前后主要差异的物质基础。结果:鉴定出了54个化学成分,PCA提示经不同方法炮制后枳壳组间差异性较大,PLS-DA筛选出VIP值1的14个化学成分作为炮制前后差异性的主要化学标记物,包括橙皮素、枸橘苷、柚皮芸香苷、柚皮苷、橙皮苷、新橙皮苷、橙皮内酯、柚皮素、去乙酰诺米林、去甲川陈皮素、橙皮素5-O-葡萄糖苷、橙皮素7-O-葡萄糖苷、新北美圣草苷和4',5'-二氢补骨脂素;将其综合加权评分后发现蜜麸枳壳中有效成分含量最高。结论:枳壳炮制前后化学成分含量发生显著变化,黄酮类成分是区分枳壳不同炮制品最重要的化合物类别,江西樟帮蜜麸枳壳为枳壳的优势炮制品种。     

冠心病不稳定心绞痛血瘀证患者尿液代谢组学特征的研究

目的利用磁共振波谱代谢学方法研究冠心病不稳定心绞痛血瘀证患者和健康志愿者尿液代谢物的变化。方法通过临床横断面调查研究,收集冠心病不稳定心绞痛血瘀证患者和健康志愿者的一般资料和尿液样品,采用磁共振(NMR)和模式识别技术对磁共振氢谱进行分析,对谱图去水峰后采用0.01化学位移分段积分和归一化处理。所得数据输入SIMCA-P软件,采用偏最小二乘-判别分析法(PLS-DA)对结果进行分析。结果冠心病不稳定心绞痛血瘀证患者尿液中脯氨酸、丙氨酸、异亮氨酸、葡萄糖、缬氨酸、组氨酸、马尿酸等物质的含量升高;柠檬酸、肌酸酐、牛磺酸等物质的含量下降。结合PLS-DA对尿液样品的分析结果显示冠心病患者与健康人在代谢组学方面存在差异,且化合物分布在性别方面也有差异。结论研究表明尿液样品中柠檬酸、脯氨酸、异亮氨酸、牛磺酸等代谢物的改变构成了冠心病不稳定性心绞痛血瘀证患者的代谢组学特征,从而为疾病的诊断和治疗以及中医证候研究提供了新思路和新方法。     

A combined HPLC-PDA and HPLC-MS method for quantitative and qualitative analysis of 10 major constituents in the traditional Chinese medicine Zuo Gui Wan

A method using a high-performance liquid chromatograph coupled with a photodiode array UV detector (HPLC-DAD) and an electrospray ionization mass spectrometer (HPLC-ESI/MS) was developed for the quality assessment (fingerprinting and simultaneous quantification of 10 major constituents) of Zuo Gui Wan (ZGW), a Traditional Chinese Medicine (TCM). The compounds were identified on the basis of comparison of their mass spectra with literature data and standard samples and quantified by the HPLC-DAD method. Baseline separation was achieved on an ODS-100V C₁₈ column (3.0 μm, 150 mm × 4.6 mm I.D.) with linear gradient elution of acetonitrile–0.1% formic acid. The method was validated for linearity (r² > 0.9996), repeatability (R.S.D. < 3.5%), intra- and inter-day precision (R.S.D. < 2.0%) with accuracy (91.8–99.7%), recovery (99.9–105.6%), limits of detection (1.4–22.0 ng), and limits of quantification (3.4–73.4 ng). The 10 compounds were selected for quality assessment of ZGW by using partial least squares-discrimination analysis (PLS-DA). The similarities of 20 batches of ZGW and their classification according to their manufacturers were based on the retention times and peak areas of the characteristic compounds.     

基于多元统计方法和顶空-气相色谱-质谱联用技术对瓜馥木不同部位挥发性成分的比较研究＊

目的 分析瓜馥木中的挥发性成分，比较根、茎、叶和虫瘿中所含挥发性成分的差异。方法 采用顶空-气相色谱-质谱（HS-GC-MS）联用技术分析瓜馥木不同部位的挥发性成分，并利用XCMS-ONLINE的组学方法和多元统计方法分析处理数据，包括偏最小二乘法判别分析（PLS-DA）、正交校正的偏最小二乘分析（OPLS-DA），辅以保留指数（RI）完成各挥发性成分的定性和半定量分析。结果 共定性出41种挥发性成分，其中根含24种成分，茎含38种成分，叶含35种成分，虫瘿含25种成分，各部位的共有成分19种。结论 瓜馥木挥发性成分主要是烯烃类和脂肪烃衍生物类，不同部位的挥发性成分有显著差异，研究结果将为该药材的质量标准科学制定和民间使用提供依据和借鉴，并为进一步的药效评价、资源开发利用奠定基础。     

Biomarkers in the early period of acute myocardial infarction in rat serum and protective effects of Shexiang Baoxin Pill using a metabolomic method

Ethnopharmacological relevance

To identify the biomarkers in early period of acute myocardial infarction (AMI) in rat serum and reveal the effective mechanism of a Traditional Chinese Medicine (TCM) named Shexiang Baoxin Pill (SBP).

Material and method

A metabolomic approach using reversed-phase liquid chromatography/quadrupole time-of-flight mass spectrometry (LC-Q-TOF-MS) was developed.

Results

Fourteen biomarkers in the early period of acute myocardial infarction (AMI) in rat serum were identified. These biomarkers include 5-methylcytosine, cystathionine ketimine, 2-oxoadipic acid, thymidine, epinephrine, homocystine, uric acid, 12(S)-hydroperoxyeicosatetraenoic acid (12s-HPETE), 11-dehydrocorticosterone, 12(S)-hydroxyeicosatetraenoic acid (12s-HETE), deoxycorticosterone, corticosterone, aldosterone and cortisol. Through pathway analysis of these biomarkers, inflammation, hypertrophy and oxidative injury were considered the most relevant pathological changes in early period of AMI.

Conclusion

Identification of AMI biomarkers not only supplied a systematic view of the progression of AMI in the early period but also provided the theoretical basis for the prevention or treatment of AMI. The results demonstrated that SBP pretreatment could offer protective effects for AMI through regulating the pathway of steroid hormone biosynthesis.     

不同产区木瓜药材质量对比分析

目的 考察市售木瓜药材主流商品质量,为木瓜药材商品规格等级标准的建立奠定基础.方法 收集三大主流产区的木瓜样品38批,采用HPLC特征图谱表征其化学成分,运用SPSS软件整体分析特征图谱相似度数据,采用偏最小二乘-判别分析(PLS-DA)特征图谱统计分析,并用正交偏最小二乘-判别分析(OPLS-DA)对宣木瓜和资丘木瓜...     

Biomarkers of Parkinson's disease and Dementia with Lewy bodies

Parkinson's disease (PD) and Dementia with Lewy bodies (DLB) are progressive and disabling neurodegenerative disorders, in which signs and symptoms overlap with each other and with other neurodegenerative conditions. Currently, diagnosis, measurement of progression, and response to therapeutic intervention rely upon clinical observation. However, there remains a critical need for validated biomarkers in each of these areas. A definitive diagnostic test would improve clinical management and enrollment into clinical trials. An objective measure of progression is vitally important in identifying neuroprotective interventions. Biomarkers may also provide insight into pathogenesis, and might therefore suggest possible novel targets for therapeutic intervention. In addition, certain biomarkers might be of use in monitoring the biochemical and physiological effects of therapeutic interventions. Development of diagnostic biomarkers has focused until recently upon imaging techniques based upon measuring loss of dopamine neurons. Additionally, advances in understanding the genetic contribution to neurodegenerative disorders, in particular in PD, have identified multiple causative genes and risk factors that in some cases may help estimate PD risk. However, recent availability of increasingly sophisticated bioinformatics technology has rendered development of fluid biomarkers feasible, opening the possibility of generally accessible blood or cerebrospinal fluid (CSF) tests that could impact upon diagnosis, management, and research in PD, PDD, and DLB.     

Quality evaluation of Angelica acutiloba Kitagawa roots by 1H NMR-based metabolic fingerprinting

The prices of Angelica acutiloba Kitagawa (yamato-toki) and A. acutiloba Kitagawa var. sukiyamae Hikino (hokkai-toki) are now mainly determined according to the sensory quality determined by experts in addition to the physical properties. This method provides a low reliability result for differentiating and qualifying their qualities. In addition, the quality in terms of pharmacological efficiency is not taken into account for consideration in the ordinary sensory method. A combination of a (1)H NMR technique and a multivariate analysis was preliminarily applied for the quality evaluation of both toki roots with regard to their geographical and variety differences. A broad range of metabolites was detected by a single-run (1)H NMR spectrometry. Partial least-squares discrimination analysis (PLS-DA), a pattern recognition method, was applied to the (1)H NMR spectra of aqueous extracts of toki samples having different sensory qualities. The PLS-DA result showed a clear clustering corresponding to the cultivation area between toki samples cultivated in Hokkaido (Japan) and those cultivated in the southern part of China and the Nara prefecture (Japan), while there was no separation corresponding to the toki's variety and sensory qualities, indicating the inconsistency of the sensory evaluation result. The chemical metabolites contributing to the discrimination of toki samples in relation to pharmacological and sensory properties were reported for the first time. A reliable multivariate calibration model used to predict the sensory quality was successfully carried out by PLS regression.     

1HNMR-偏最小二乘法-判别法分析椰奶的质量

目的 建立一种基于核磁共振-模式识别技术来鉴别椰奶质量的新方法.方法 利用1HNMR测定椰奶样品,采集信息并转化成数据矩阵;利用模式识别方法中的PLS-DA法对数据进行分析,并建立鉴别模型.结果 5个厂家的椰奶样品均能获得有效的鉴别,含有添加物的椰奶样品也能有效地与正常椰奶样品鉴别开来.结论 所用方法可靠、快速,能有效地鉴别椰奶的品质.