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Zhang S.,Nanchang University | Liu Q.,Nanchang University | Xiao J.,Jiangxi Provincial Maternal and Child Health Hospital | Lei J.,Jiangxi Childrens Hospital | And 3 more authors.
International Immunopharmacology | Year: 2016

Background The precision-cut kidney slice (PCKS) model appears to be a useful ex vivo model of renal fibrosis. However, little in-depth molecular investigation on the PCKS model has been performed. Therefore, the aim of this study will be to investigate and validate the molecular validity of this model. Methods The PCKS model was constructed in male C57BL/6 mice. To induce renal fibrosis, PCKS were incubated in recombinant human TGF-β1 for 48 h. Protein expression of phosphorylated Smad2 (p-Smad2, cytosolic and nuclear), Smad7, phosphorylated ERK1 (p-ERK1), phosphorylated ERK2 (p-ERK2), and phosphorylated p38 MAPK (p-p38 MAPK) was measured using Western blotting. To assess Smad2/3 heteromeric complex formation and phosphorylated Smad3 (p-Smad3) expression, immunoprecipitation was performed with an anti-Smad2 or an anti-Smad3 antibody, respectively, prior to Western blotting. Results p-Smad2 and p-Smad3 were significantly upregulated in the PCKS model relative to control (p < 0.05). However, we found no significant difference in Smad7 expression between the PCKS model and control (p > 0.05). The PCKS model demonstrated significantly greater Smad2/3 complex formation and nuclear translocation relative to control (p < 0.05). The PCKS model showed significantly greater expression of p-ERK1, p-ERK2, and p-p38 MAPK relative to control (p < 0.05). Conclusions The PCKS model displays several well-established molecular markers of renal fibrosis. However, the PCKS model failed to display Smad7 downregulation and appears to display “over-activation” of p-Smad2 and p-Smad3 as well as “under-activation” of ERK1/2 and p38 MAPK signaling vis-à-vis the well-established in vivo unilateral ureteric obstruction model of renal fibrosis. © 2016

Liu X.,Shanghai Tobacco Group Corporation Ltd | Zhang J.,Shanghai Tobacco Group Corporation Ltd | Zhang C.,Shanghai Tobacco Group Corporation Ltd | Yang B.,Jiangxi Provincial Maternal and Child Health Hospital | And 3 more authors.
Toxicology Research | Year: 2016

4-(Methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK) is considered to be the most carcinogenic of the four tobacco-specific nitrosamines (TSNAs) and it needs to be metabolically activated to exert its carcinogenic effect on humans. For the simultaneous intake of NNK and other compounds with similar molecular structures in the context of tobacco smoke, whether (R,S)-N-nitrosoanatabine (NAT), (R,S)-N-nitrosoanabasine (NAB) and nicotine contribute to the inhibitory potency of the cytochrome P450 (CYP) enzyme-catalyzed NNK metabolism or not needs to be investigated. In the in vitro study, 4-oxo-4-(3-pyridyl) butanal (OPB), 4-hydroxy-1-(3-pyridyl)-1-butanone (HPB) and 4-oxo-4-(3-pyridyl) butanoic acid (OPBA) were established as the products of the CYP2A13-catalyzed NNK metabolism and the kinetic parameters were calculated from the Michaelis-Menten equation. Addition of NAT, NAB or nicotine resulted in a competitive inhibition for the NNK metabolism catalyzed by CYP2A13. The inhibition constant Ki values were calculated to be 0.21 μM (NAT), 0.23 μM (NAB) and 8.51 μM (nicotine) for OPB formation; 0.71 μM (NAT), 0.87 μM (NAB) and 25.01 μM (nicotine) for HPB formation and 0.36 μM (NAT), 0.50 μM (NAB) and 6.57 μM (nicotine) for OPBA formation, respectively. In addition, the study of the transformation of the three metabolites revealed OPB was not only an end product but also an intermediate product of the CYP2A13-catalyzed NNK metabolism. These results suggest that structurally similar tobacco constituents with weak or no carcinogenicity influence the metabolic activation of NNK, which interferes with its carcinogenicity to some extent. © The Royal Society of Chemistry 2016.

Yang B.-C.,Jiangxi Provincial Maternal and Child Health Hospital | Liu F.-Y.,Jiangxi Provincial Maternal and Child Health Hospital | Wang L.-Q.,Jiangxi Provincial Maternal and Child Health Hospital | Zou Y.,Jiangxi Provincial Maternal and Child Health Hospital | And 6 more authors.
Analytical Methods | Year: 2015

A high throughput metabolite fingerprinting tool based on wooden-tip electrospray ionization mass spectrometry (WT-ESI-MS) has been established for the serum metabolic profiling study of endometriosis with little sample pre-treatment, no chromatography and instrument cycle times of less than 5 min. Serum samples from endometriosis patients and healthy controls were analyzed by direct WT-ESI-MS with a high resolution ESI-Q-TOF-MS. The resulting data were analyzed by multivariate data analysis. MS/MS experiments were carried out to identify potential biomarkers. Global metabolic profiling and subsequent multivariate analysis clearly distinguished endometriosis patients from healthy controls. A total of ten metabolites, up-regulated or down-regulated, were identified which contribute to the progress of endometriosis. These promising identified biomarkers underpin the metabolic pathway including steroid hormone biosynthesis, glycerophospholipid metabolism, sphingolipid metabolism, pyruvate metabolism, bile acid biosynthesis, and androgen and estrogen metabolisms. Considering that a much higher throughput can be obtained without a chromatographic step, the present WT-ESI-MS method could be developed as a fast prognostic or diagnostic method for endometriosis. This journal is © The Royal Society of Chemistry 2015.

Yang B.-C.,Jiangxi Provincial Maternal and Child Health Hospital | Wang F.,Jiangxi Provincial Maternal and Child Health Hospital | Deng W.,Jiangxi Provincial Maternal and Child Health Hospital | Zou Y.,Jiangxi Provincial Maternal and Child Health Hospital | And 5 more authors.
Analytical Methods | Year: 2015

Wooden-tip electrospray ionization (wooden-tip ESI) is applied to the rapid, in situ, direct qualitative and quantitative trace analysis of toxic and hazardous compounds in food samples. To evaluate the potential of wooden-tip ESI mass spectrometry (MS) in food analysis, pesticides, toxicants, date-rape drugs, and illicit additives were detected in various food samples. Wooden-tip ESI-MS experiments were performed using wooden tip sampling and ionization, with examination using MS and tandem mass spectrometry (MS/MS). The results proved that wooden-tip ESI allows the detection and confirmation of traces of toxic and hazardous compounds in food. In addition, selected analytes in beverages were obtained at absolute levels as low as ∼10 pg. Quantitation of analytes in liquid and powder samples was also evaluated. Single sample analysis was completed within 2 min. The data obtained shows that the wooden-tip ESI-MS is a promising tool for rapid analysis of food samples. © The Royal Society of Chemistry 2015.

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