5.A HPLC-Q-TOF-MS-based urinary metabolomic approach to identification of potential biomarkers of metabolic syndrome.
Zhi-Rui, YU ; Yu, NING ; Hao, YU ; Nai-Jun, TANG
Journal of Huazhong University of Science and Technology (Medical Sciences) 2014;34(2):276-83
Metabolic syndrome (MetS) is a serious threat to public health worldwide with an increased risk of developing type 2 diabetes, cardiovascular diseases and all-cause morbidity and mortality. In this study, a urinary metabolomic approach was performed on high performance liquid chromatography quadrupole time-of-flight mass spectrometry to discriminate 36 male MetS patients and 36 sex and age matched healthy controls. Pattern recognition analyses (principal component analysis and orthogonal projections to latent structures discriminate analysis) commonly demonstrated the difference between MetS patients and no-MetS subjects. This study found 8 metabolites that showed significant changes in patients with MetS, including branch-chain and aromatic amino acids (leucine, tyrosine, phenylalanine and tryptophan), short-chain acylcanitine (tiglylcarnitine), tricarboxylic acid (TCA) cycle intermediate (cis-aconitic acid) and glucuronidated products (cortolone-3-glucuronide and tetrahydroaldosterone-3-glucuronide). The candidate biomarkers revealed in this study could be useful in providing clues for further research focusing on the in-depth investigation of the cause of and cure for MetS.
6.Induction of aryl hydrocarbon receptor and CYP1A1 mRNA by 2,3,7,8-tetrachlorodibenzo-p-dioxin in rat liver.
Yun-ru LIU ; Nai-jun TANG ; Da-lin REN
Chinese Journal of Industrial Hygiene and Occupational Diseases 2003;21(6):417-419
OBJECTIVETo explore the toxic mechanism of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) by studying the induction of cytochrome P4501A1 (CYP1A1) and aryl hydrocarbon receptor (AHR) mRNA in liver of TCDD-treated SD rats.
METHODSThirty female SD rats were randomly divided into control group and 5 exposure groups, every group had 5 rats. The animals were treated i.p. with 0.01, 0.1, 1, 10, 50 microg TCDD/kg BW. AHR and CYP1A1 mRNA expression were analyzed by RT-PCR after 24 h.
RESULTSThe contents of AHR and CYP1A1 mRNA were increased in all exposure groups except the 0.01 microg TCDD/kg BW group. AHR mRNA content was significantly increased in 50 microg TCDD/kg BW group (P<0.05); CYP1A1 mRNA contents were significantly increased in all exposure groups (P<0.05) but not 0.01 microg TCDD/kg BW group. There were dose-response relationship between TCDD doses and AHR, CYP1A1 gene expression.
CONCLUSIONBoth AHR and CYP1A1 gene in liver of TCDD-treated SD rats can be induced 24 h after exposure and CYP1A1 gene is more inducible than AHR gene.
Animals ; Cytochrome P-450 CYP1A1 ; genetics ; Dose-Response Relationship, Drug ; Female ; Gene Expression Regulation ; drug effects ; Liver ; drug effects ; metabolism ; Polychlorinated Dibenzodioxins ; toxicity ; Rats ; Rats, Sprague-Dawley ; Receptors, Aryl Hydrocarbon ; genetics
7.Preventive effect of TGF-?_1 neutralizing antibody on flexor tendon adhesion from operation
Jin-Tang WANG ; Yin-Gang ZHANG ; Jun LI ; Nai-Gui BI ; Xue-Zhe HAN ; Xiao-Wei ZHANG ;
Chinese Journal of Microsurgery 2006;0(05):-
Objective To investigate the preventive effect of TGF-?_1 neutralizing antibody on flexor tendon adhesion from operation.Methods One hundred and eight leghon cocks performed anastomonsis op- eration were divieded into three groups randomly,as normal saline(control group),5?g/ml group,10?g/ml group of TGF-?_1,antibody.At 1 st,3rd,8th and 12th weeks respectively after operation,the flexor biomechan- ics test,HE staining,Masson staining,Sirius red-polarization staining and TGF-?_1 immunohistochemistry stai- ning were used.Results The max of strength of tendon and the stimulate active flexor from the experiment groups(5?g/ml group,10?g/ml) are higher than from the control group,The max of strength of tendon of the experiment groups are less at 8th weeks,and no difference at 12th weeks from the control group;Compared with the control group,the 10?g/ml group were less shorten the progress of inflammation and accelerated the progress of molding;In the experiment groups(5?g/ml group,10?g/ml),the density of the collagenⅠtype were less,the ratio ofⅠ/Ⅲcollagen and expression of the TGF-?_1 were decreasing.Condusion The study showed that applying of TGF-?_1 muhiclonal neutralizing antibody can inhibit efficiently the function of the TGF-?_1 during the flexor tendon repair,reduce tendon adhesion and scar fromation,however has no affec- tion of tendon intensity,suggesting it is a latent and efficient method for preventiong flexor tendon from adhe- ring after operation.
8.Construction and primary application of oligonucleotide microarray specialized for pancreatic adenocarcinoma-associated genes.
Xin SHI ; Wen-jun WEI ; Nai-rong GAO ; Zhang-jun CHENG ; Yong-hui TANG
Chinese Journal of Surgery 2007;45(1):39-42
OBJECTIVETo investigate the construction of oligonucleotide microarray specialized for pancreatic adenocarcinoma-associated genes and its application.
METHODSPancreatic cancer related genes were purposely selected, and oligonucleotide microarray was prepared by spotting oligonucleotide probes onto glass slides coated with APS-PDC. Total RNA were extracted from frozen tissues with TRIzol method according to the manufacturer's protocol, and purified with QIAGEN RNeasy Kit. Labeled cDNA targets for hybridizations were synthesized by reverse transcription from control- and cancer-total RNA samples in the presence of Cy5-dCTP and Cy3-dCTP, respectively. The labeled probes were hybridized with oligonucleotide microarray for 16 h to 18 h. Hybridized microarray was scanned by Agilent laser scanner, and the acquired image was analyzed by Imagene3.0 software. The intensity ratio of Cy3 and Cy5 were calculated. To confirm the expression profiles of these genes, quantitative reverse transcription-PCR (Q RT-PCR) was carried out with CDC25B and TUSC3 genes. The product of PCR were quantitated by comparative Ct method.
RESULTSThe signal of microarray hybridization was clear, and the images had a lower background and higher signal-noise ratio. The signal of positive control spots were uniform, and spots of negative control and blank signal were fairly low. In comparison with normal pancreas, 24 differential expressed genes were identified, which included 17 up-regulated and 7 down-regulated genes. The results of Q RT-PCR demonstrated that the expression of CDC25B and TUSC3 in pancreatic cancer were increased and decreased respectively, which consistent with microarray hybridization.
CONCLUSIONSThe oligonucleotide microarray specialized for pancreatic cancer are desirable for its specialty, flexibility and sensitivity, which can simultaneously and parallelly detect multiple pancreatic cancer-associated genes. In contrast to normal pancreatic tissues, the genes expression profile are different significantly in pancreatic cancer.
Adenocarcinoma ; genetics ; pathology ; Aged ; Female ; Gene Expression Profiling ; Humans ; Male ; Middle Aged ; Oligonucleotide Array Sequence Analysis ; methods ; Oligonucleotide Probes ; Pancreatic Neoplasms ; genetics ; pathology ; Reproducibility of Results ; Reverse Transcriptase Polymerase Chain Reaction ; methods
9.A HPLC-Q-TOF-MS-based urinary metabolomic approach to identification of potential biomarkers of metabolic syndrome.
Zhi-rui YU ; Yu NING ; Hao YU ; Nai-jun TANG
Journal of Huazhong University of Science and Technology (Medical Sciences) 2014;34(2):276-283
Metabolic syndrome (MetS) is a serious threat to public health worldwide with an increased risk of developing type 2 diabetes, cardiovascular diseases and all-cause morbidity and mortality. In this study, a urinary metabolomic approach was performed on high performance liquid chromatography quadrupole time-of-flight mass spectrometry to discriminate 36 male MetS patients and 36 sex and age matched healthy controls. Pattern recognition analyses (principal component analysis and orthogonal projections to latent structures discriminate analysis) commonly demonstrated the difference between MetS patients and no-MetS subjects. This study found 8 metabolites that showed significant changes in patients with MetS, including branch-chain and aromatic amino acids (leucine, tyrosine, phenylalanine and tryptophan), short-chain acylcanitine (tiglylcarnitine), tricarboxylic acid (TCA) cycle intermediate (cis-aconitic acid) and glucuronidated products (cortolone-3-glucuronide and tetrahydroaldosterone-3-glucuronide). The candidate biomarkers revealed in this study could be useful in providing clues for further research focusing on the in-depth investigation of the cause of and cure for MetS.
Adult
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Biomarkers
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urine
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Chromatography, High Pressure Liquid
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Female
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Humans
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Male
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Mass Spectrometry
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Metabolic Syndrome
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diagnosis
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urine
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Metabolomics
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Middle Aged
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Principal Component Analysis
10.Effects of dibutyl phthalate and monobutyl phthalate on testosterone secretion and insulin-like factor 3 expression of Leydig tumor cells in mice.
Qing-hong ZHOU ; Xi CHEN ; Ling LENG ; Jing-shu ZHANG ; Nai-jun TANG
Chinese Journal of Industrial Hygiene and Occupational Diseases 2013;31(2):83-87
OBJECTIVETo observe the effects of dibutyl phthalate (DBP) and monobutyl phthalate (MBP) on the mRNA and protein expression of insulin-like factor 3 (INSL3) in the Leydig tumor cells (MA-10) of mice and the level of testosterone secreted from MA-10 cells.
METHODSThe MA-10 cells of mice, used as a cellular model, were exposed to DBP and MBP. The content of testosterone in the supernatant medium was measured by enzyme-linked immunosorbent assay; the mRNA and protein expression levels of INSL3 in MA-10 cells were measured by quantitative PCR and Western Blot.
RESULTSCompared with the control group, MA-10 cells showed increased synthesis of testosterone when exposed to low concentrations of DBP and MBP (10(-9) ∼ 10(-6) mol/L) and inhibited synthesis of testosterone when exposed to high concentrations of DBP and MBP (10(-3) mol/L), and the typical two-way effects became more significant as the time went one and the concentrations increased (P < 0.05). Compared with the control group, MA-10 cells showed significantly lower mRNA and protein expression levels of INSL3 when exposed to 10(-6) and 10(-4) mol/L DBP (P < 0.05); MA-10 cells showed increased protein expression of INSL3 when exposed to 10(-7) mol/L MBP, and the mRNA and protein expression levels of INSL3 decreased as the concentration of MBP increased.
CONCLUSIONDBP and MBP can inhibit the secretion of testosterone from MA-10 cells at high concentrations, but stimulate the secretion of testosterone at low concentrations. Both DBP and MBP have inhibitory effects on the mRNA and protein expression of INSL3 in MA-10 cells.
Animals ; Cell Line ; Dibutyl Phthalate ; toxicity ; Insulin ; metabolism ; Leydig Cells ; drug effects ; metabolism ; Male ; Mice ; Phthalic Acids ; toxicity ; Proteins ; metabolism ; Testosterone ; secretion