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', 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.

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.

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.

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.

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.

Objective To investigate the roles of TLR3 pathway activiated by polyI:C in proliferation and apoptosis of multiple myeloma (MM) RPMI8226 cell line.Methods RPMI8226 cells were cultured in RPMI 1640 with different dose of polyl:C.Cells were collected in different time.Proliferation and apoptosis were detected by CCK-8 kit and flow cytometry,separately.Results The proliferation of RPM18226 was inhibited by polyI:C,and it was dose and time dependent,24 h:12.30％ ±2.04％,22.50％±2.20％,37.90％ ±1.30％ ; 48 h:17.80％ ±1.52％,29.60％ ±0.85％,45.80％ ±1.68％ ;72 h:25.10％±1.01％,34.60％±1.27％,60.50％±2.08％,P＜0.05.RPMI8226 cells were incubated with 50 μg/ml,100 μg/ml and 200 μg/ml polyI:C for 48 h.Apoptotic rate were 5.60％ ±1.06％,8.71％ ±1.06％ and 13.93％ ±1.17％,P＜0.05.TLR3 and TRIF mRNA expression increased obviously and dose dependent,TLR3:1.41±0.10,2.24±0.16,4.08±0.13; TRIF:1.07±0.16,1.97±0.13,3.56±0.19,P＜0.05.Conclusion The proliferation of MM cells were inhibited by TLR3 pathway obviously,and apoptosis was induced by polyI:C.

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.

Objective To explore the surgical management of chronic pancreatitis complicated with pancreatolithiasis (CPPL). Methods The clinical data of 66 patients with CPPL were retrospectively analyzed. Pancreatolithiasis was classified into 4 types according to the location of stones: stones located in the head of the pancreas (type Ⅰ, n=28), stones located in the body of the pancreas (type Ⅱ, n=30), stones located in the tail of the pancreas (type Ⅲ, n=1) and stones located from the head to tail of the main duct of pancreas (type Ⅳ, n=7). Ten patients (including 4 with type Ⅰpancreatolithiasis, 5 with type Ⅱ and 1 with type Ⅳ) received conservative treatment; 10 patients with type Ⅰ pancreatolithiasis underwent lithotomy under endoscope; pancreaticoduodenectomy and Beger procedure were carried out on 14 patients with type Ⅰ pancreatolithiasis, pancreatolithotomy+pancreaticojejunostomy on 25 patients with type Ⅱ pancreatolithiasis, resection of pancreatic tail and spleen on 1 patient with type Ⅲ pancreatolithiasis, and Puestow-Gillesby procedure, dividing of the neck of pancreas+removing stones from both ends of pancreatic duct+Roux-en-Y pancreatojejunostomy on 6 patients with type Ⅳ pancreatolithiasis. Results Sixty-two patients were followed up for 2 months to 15 years, and the number of patients with recurrence for type Ⅰ, Ⅱ, Ⅲ and Ⅳ pancreatolithiasis was 4, 2, 0 and 3, respectively. Conclusions Early surgical management according to the location of stones should be carried out after confirmed diagnosis of CPPL. Individualized management based on correct diagnosis and classification plays an important role in the prevention of pancreatolithiasis recurrence.