1.Quantitative metabolomics based on NMR.
Chunying JIANG ; Yinghong WANG
Acta Pharmaceutica Sinica 2014;49(7):949-55
Nuclear magnetic resonance (NMR) spectroscopy can be used to both identify and quantify chemicals from complex mixtures. Over the last several decades, significant technical and experimental advances have made quantitative nuclear magnetic resonance (qNMR) a valuable analytical tool for quantitative measurements of a wide variety of samples. This particular approach is now being exploited to characterize the metabolomes of many different biological samples and is called quantitative metabolomics or targeted metabolic profiling. In this review, some of the strengths, limitations of NMR-based quantitative metabolomics will be discussed as well as the practical considerations necessary for acquisition with an emphasis on their use for bioanalysis. Recent examples of the application of this particular approach to metabolomics studies will be also presented.
2.LOXL2 and its relation with epithelial-mesenchymal transition cholangiocarcinoma
Yinghong GAO ; Tianyu LI ; Zhanfeng GAO ; Weiwei JIANG ; Shuguang WANG
Chinese Journal of General Surgery 2008;23(10):784-787
Objective To investigate the expression of LOXL2 protein (lysyl oxidase like-2 protein) and epithelial-mesenchymal transition (EMT) related markers in cholangiocarcinoma tissues and its relation with the malignant features. Methods The expression of LOXL2、E-cadherin and Vimentin protein in 48 cases of cholangiocarcinoma tissues was detected by immunohistochemistry and compared with the clinicopathological data of cholangiocarcinoma. Results The positive expression rate in cholangiocarcinoma was 71% ( 34/48 ) for LOXL2 and 46% ( 22/48 ) for Vimentin, the absent expression rate was 52% (25/48) for E-cadherin. The positive expression rate of LOXL2 was significantly associated with the absent expression of epithelium markers E-cadherin ( r = 0. 394, P < 0. 05 ) and the positive expression of fibroblast markers Vimentin ( r = 0. 406, P < 0. 05 ). There was no correlation between the expression of LOXL2 and patients gender, age, and cancer differentiation, but a significant correlation with tumor metastasis was found ( P < 0. 05 ). Conclusions LOXL2 protein overexpression in cholangiocarcinoma may accelerate invasion of cholangiocarcinoma through induced EMT.
4.Evaluation of dose-related effects of 2', 3', 5'-tri-O-acetyl-N6-(3-hydroxylaniline)adenosine using NMR-based metabolomics.
Zhaoxia MIAO ; Liu YANG ; Chunying JIANG ; Yinghong WANG ; Haibo ZHU
Acta Pharmaceutica Sinica 2014;49(5):679-85
2', 3', 5'-Tri-O-acetyl-N6-(3-hydroxylaniline)adenosine (WS070117) is a derivative compound of natural product cordycepin. It has significant lipids regulating activity and low toxicity which has been proved by in vitro and in vivo experiments. In this study, 1H NMR-based metabolomics was used to investigate the dose-related effects of WS070117 on hyperlipidemia of high-fat-fed hamsters. The hyperlipidemic hamsters were administrated with six different doses of WS070117, including 3, 12, 50, 100, 200 and 400 mg x kg(-1) x d(-1). 1H NMR spectra of hamster serum were visually and statistically analyzed using two multivariate analyses: principal component analysis (PCA) and orthogonal partial least squares-discriminant analysis (OPLS-DA). As a result, WS070117-treated groups showed dose-related regulation of metabolites associated with lipid metabolism, choline metabolism and glucose metabolism. The dose of 3 mg x kg(-1) x d(-1) of WS070117 only exhibited a little lipids regulating activity. However, the doses of 12 and 50 mg x kg(-1) x d(-1) of WS070117 both regulated the contents of metabolites to reverse significantly toward normal levels. When the dose of WS070117 reached 100 mg x kg(-1) x d(-1), it was more effective than positive control drugs. The work suggested that NMR-based metabolomics might be a valuable approach to evaluate dose-related effects of lipids regulating compounds.
5.Function Study of the Nano-Molecular PAMAM as miR Carrier Targeting Gastric Adenocarcinoma
Wei SONG ; Zhongmin JIANG ; Ping XU ; Yinghong LIANG ; Xiaozhi LIU
Tianjin Medical Journal 2013;(12):1147-1149
Objective To study the function of the nano-molecular polyamide-amine (PAMAM) as microRNA(miR) carrier targeting gastric adenocarcinoma, and the foundation of developing an efficient delivery of small molecule drugs tar-geting gastric cancer thereof. Methods The folic acid (FA)/PAMAM comoles compound was prepared by dialysis method. After transfection of miR-7 or liposomes into SGC-7901 cell line, fluorescence microscope was used to detect the gene trans-fect efficiency. Real-time quantitative polymerase chain reaction (qRT-PCR) was used to detect the miR-7 level. The immu-nocytochemistry assay was used to test the protein expressions of epidermal growth factor receptor (EGFR), protein kinase B (PKB) and proliferating cell nuclearantigen (PCNA). The transwell system was utilized to explore the migration ability of tu-mor cells. Results Compared with liposme, FA/PAMAN complex compound can significantly improve the level of miR-7 in SGC-7901 cells,reduce the protein levels of EGFR, PKB and PCNA in SGC-7901 cells, and also reduce the percentage of cancer cell migration (P<0.05). Conclusion PAMAM can effectively transfect miR into gastric cancer cells, which is expected to become an efficient delivery of small molecule drugs.
6.Glycoprotein I_(1-2-1) from Irpex lacteus
Zhenwei YANG ; Ruizhi JIANG ; Yinghong CHEN ; Qipin GAO
Chinese Traditional and Herbal Drugs 1994;0(08):-
Objective To study the chemical structure of a pure glycoprotein I_(1-2-1) from Irpex lacteus.Methods Based on chemical and spectral analysis,the structural characterization of I_(1-2-1) was investigated.Results I_(1-2-1) was composed of Ara,Xyl,Man,Gal,and Glu with its mean molecular weight of(40 000.) Methylation analysis showed that the main chain of I_(1-2-1)was all 1→2,1→6 linked Manp.Conclusion I_(1-2-1) is a complicated glycoprotein obtained for the first time from I.lacteus.
7.Analysis and quantitative determination of fatty acids in Hericium Erinaceus polysaccharide
Yongjie YANG ; Ruizhi JIANG ; Yinghong CHEN ; Qipin GAO
Chinese Traditional Patent Medicine 1992;0(10):-
AIM: To assay fatty acids in Hericium Erinaceus polysaccharide by GC for establishing a method for quantitative determination of fatty acids. METHODS: The kinds of fatty acids were detected by GC-MS. The fatty acids contents were determined by GC: The sample was dissolved in sulphuric acid-methanol (1∶20) containing 0.5 mg heptadecanoic acid methyl ester and kept at 80?C for 3 hr. Then, the solution was extracted by hexamethylene four times and diluted to 1 mL. The extraction (0.4 ?L) was injected on a DB-1 column, using nitrogen as the carrier gas. The temperature was raised from 60?C to 200?C at the rate of 10?C?min -1 , and subsequently sustained for 30 min. The injection port and detector temperature were both at 250?C. RESULTS: The fatty acids were separated completely. The linear ranges were 0.4-3.6 mg?mL -1 . RSD97%. CONCLUSION: The fatty acids can be determined accurately by this method.
8.Expression of B1a cells and IgA1 positive cells in tonsil of IgA nephropathy patients and analysis of associated clinicopathological factors
Qidi HOU ; Gang WU ; Youming PENG ; Hong LIU ; Yinghong LIU ; Xiangqing XU ; Wenling JIANG ; Fuyou LIU
Chinese Journal of Nephrology 2011;27(7):475-478
Objective To examine the expression of IgA1 and B1a positive cells in palatine tonsils of IgA nephropathy (IgAN) patients, and to analyze the association between B1a cells and clinicopathological changes. Methods Eight patients diagnosed as IgAN by renal biopsy and 8 chronic tonsillitis patients without nephritis as control were enrolled in the study.Immunofluorescence and laser scanning confocal microscope (LSCM) were applied to observe the localization and quantitative calculation of Bla and IgA1 positive cells. Statistic analysis of the association of B1a cells with proteinuria and pathological Lee's grading was performed. Results Bla cells were mainly localized in germinal center of tonsil, and IgA1 positive cells were mainly localized in subepithelium of tonsil. Compared to control group, the percent of B1a cells and IgA1 positive cells was significantly higher in IgAN (P<0.01). There was a positive correlation between Bla cells and IgA1 cells (P<0.05). In IgAN, the percent of B1a cells in patients with hematuria and proteinuria was obviously higher than that of patients with hematuria only (P<0.05). The number of Bla cells in IgAN patients with≥Lee's grade Ⅲ was significantly higher than that of those < grade Ⅲ (P<0.05). Conclusions IgA1 may be secreted by Bla cells in the tonsil of IgAN patients. The number of B1a cells is correlated with exacerbation of proteinuria and pathological severity, which may play an important role in pathogenesis of IgAN.
9.The Characteristic Analysis of Astigmatism in Preschool Children in Kunming
Yinghong KANG ; Lihong LI ; Na LI ; Junmei DONG ; Yi JIANG ; Jun LU
Journal of Kunming Medical University 2013;(11):105-107
Objective To explore the types, degree and distribution regularities of axial direction in astigmatism preschool children and the relationship of astigmatism and amblyopia. Methods Mydriasis optometry was performed for preschool children in our hospital outpatient reception from 2010 to 2011,and the astigmatism in 831 eyes of 477 cases were statistically analyzed. Results The compound hyperopic astigmatism was the most, accounting for 65.8%,followed by mixed astigmatism (13.8%),simple hyperopia astigmatism (9%),compound myopia astigmatism (8.7%) and simple myopia astigmatism (2.3%). Preschool children astigmatism degree distribution was distributed among 0.50 D and 2.00 D, accounting for 42%. For astigmatism axial, the astigmatism with rule, astigmatism against the rule and oblique astigmatism was accounted for 95.9%, 2.4% and 1.7%, respectively. Conclusion Compound hyperopic astigmatism is a main type of astigmatism in preschool children, and astigmatism is closely related to the formation of amblyopia.
10.A 1H NMR based metabonomics approach to progression of coronary atherosclerosis in a hamster model.
Chunying JIANG ; Kangmin YANG ; Liu YANG ; Zhaoxia MIAO ; Yinghong WANG ; Haibo ZHU
Acta Pharmaceutica Sinica 2013;48(4):495-502
To obtain a better understanding of the progression of atherosclerosis and identify potential biomarkers, proton nuclear magnetic resonance spectroscopy (1H NMR)-based metabonomics was used to study the metabolic changes in the plasma of hamster fed with a high-fat/cholesterol diet. Plasma samples were collected at different time points during the progression of atherosclerosis and individual proton NMR spectra were visually and statistically assessed using multivariate analyses. NMR results for all samples showed a time-dependent development from physiological to pathophysiological status during atherosclerosis. Analysis of the identified biomarkers of atherosclerosis suggests that lipid and amino acid metabolisms are significantly disturbed, together with inflammation, oxidative stress, following cholesterol overloading. The results enriched our understanding of the mechanism of atherosclerosis and demonstrated the effectiveness of the NMR-based metabonomics approach to study such a complex disease.