Effect of Dry-cold Environment on Metabolic Components in Urine and Plasma of Rats / 中国实验方剂学杂志

Objective:Nuclear magnetic resonance hydrogen spectrum(1H-NMR) was adopted to investigate the effect of dry-cold environment and diet on urine and plasma metabolites of rats,in order to find possible biomarkers and their related metabolic pathways. Method:The dry-cold environment and diet were used as a stressors to intervene rats for 3 weeks,urine and plasma samples were collected.1H-NMR combined with multivariate data analysis were used to identify relevant metabolic markers and analyze their metabolic pathways. Result:The dry-cold environment and diet resulted in significant changes of 23 metabolites in urine and 15 metabolites in plasma;these metabolites were closely related to changes of various metabolic pathways,namely the tricarboxylic acid(TCA) cycle,pyruvate metabolism,glycolysis or gluconeogenesis,valine,leucine and isoleucine biosynthesis,glycine,serine and threonine metabolism,histidine metabolism;glyoxylic acid and dicarboxylic acid metabolism,alanine,aspartic acid and glutamate metabolism. Conclusion:Dry-cold environment and diet conditions can lead to the changes of various metabolic pathways in the body.The metabolomics based on 1H-NMR has the advantage of explaining the pathophysiological dynamics and overall changes of the body,which is of great significance for exploring the pathogenesis of non-deterministic disease factors.

Effect of diethylnitrosamine on renal metabolic pathway in rats / 中南大学学报(医学版)

To explore the renal metabolic markers relavant to the renal toxicity of diethylnitrosamine and the metabolic pathways involved in the renal metabolic markers.
 Methods: Nineteen Sprague Dawley rats were assigned into 2 groups: A normal control group (n=9) and a diethylnitrosamine (DEN) administration group (n=10). The rats in the normal control group were given sterilized water for free drinking. The rats in the DEN administration group were given 0.1 mg/mL DEN solution for free drinking. After 18 weeks, the kidney tissues were collected and tested for nuclear magnetic resonance detection and pathological examination.
 Results: The content of kidneys metabolites in the rats with the DEN administration was changed significantly. The levels of alanine, taurine, pyruvate, acetate, and choline were significantly reduced compared with rat in the normal control group, while the levels of creatine, glycine, TMAO, methionine, proline, lactate, valine, leucine and isoleucine were significantly increased.
 Conclusion: Metabolicomics studies have revealed significant differences in five metabolic pathways, including valine, leucine and isoleucine biosynthesis, glycine serine and threonine metabolism, pyruvate metabolism, glycolysis or gluconeogenesis, cysteine and methionine metabolism.

Urine metabonomics study of abnormal Savda syndrome rat model in Uyghur traditional medicine / 中国实验动物学报

Objective To evaluate and discuss the changes of biomarkers of abnormal Savda syndrome rat model in Uyghur traditional medicine by external feature observation and urine metabolomics assessment. Methods The abnormal Savda syndrome rat model was established according to the theory of Uyghur traditional medicine. Its external characteristics such as hair, tongue, sleep, feces, emotion and weight growth rate were observed and scored, and their urine was detected and analyzed by proton nuclear magnetic resonance(1H-NMR)spectroscopy. Results Compared with the healthy control group,there were significant changes in the external features in the abnormal Savda syndrome rat model group,including dry and hard stools,reduced urine output and darker color,dry fur,dark purple tongue with ecchymosis, and decreased weight growth rate. Moreover,23 urinary metabolites were significantly reduced,including propionate,lactic acid, pyruvic acid, acetic acid, alanine, acetamide, glycoprotein, acetone, methyl guanidine, sarcosine, ornithine, glycine, creatine, creatinine, aminoanhydride, β-galactose, urocanate, tyrosine, phenylalanine, hippuric acid, aminohippuric acid,formic acid and lysine. However,urea,citric acid,allantoin and α-ketoglutaric acid were significantly increased. Conclusions During the development process of Savda syndrome, there are not only abnormal changes in external appearance in the model rats, but also evident changes of many internal metabolic pathways. The obvious abnormalities of the urine metabolites may be related to the biological mechanisms of abnormal Savda syndrome.

Plasma metabonomic study of abnormal Savda rat model of type 2 diabetes / 中国比较医学杂志

Objective To study the serum metabonomic changes in abnormal Savda rat model of type 2 diabetes mellitus (T2DM).Methods Sprague Dawley (SD) rats were subjected to abnormal Savda-producing conditions followed by induction of T2DM.Plasma samples were collected from each test group and analyzed by 1H nuclear magnetic resonance (NMR) spectroscopy.The spectral profiles were analyzed by multivariate analysis using orthogonal projection to latent structurs discriminant analysis (OPLS-DA).Results The content of glucose and lactic acid in plasma of the rats with abnormal Savda syndrome was decreased, while the contents of VLDL, LDL and carnitine increased obviously.For abnormal Savda rat model with T2DM, the concentration of amino acids (alanine, valine, isoleucine, leucine, tyrosine, glutamic acid, phenylalanine, methyl-histidine, glycine), and lactic acid, pyruvic acid, glycoprotein, citric acid, creatinine, β-hydroxybutyric acid, acetoacetic acid, acetic acid, acetate, creatine, carnitine, scyllo-inositol, VLDL, LDL were decreased, and glucose and taurine concentrations were increased significantly.Conclusions Protein metabolism, phenylalanine and tyrosine metabolic pathway, glycolytic pathway and the lipid metabolic pathway may be considered as biomarkers of abnormal Savda rat model of T2DM.

Plasma amino acid profiling of "same symptom for different disease" in Uyghur medicine based on high-performance liquid chromatography / 中南大学学报(医学版)

OBJECTIVE@#To determine the plasma amino acid metabolism of "same symptom for different diseases" in different cancer patients in Uyghur medicine.@*METHODS@#Plasma amino acid concentration was tested by high-performance liquid chromatography (HPLC) in cancer patients with different symptom, and the spectral profiles were subjected to a t-test for statistical significance.@*RESULTS@#Compared with the healthy group, lung cancer, cervical cancer, breast cancer and gastric cancer patients with abnormal Savda had lower concentration of plasma amino acids except some amino acids. Lung cancer patients with abnormal Savda had higher concentration of plasma phenylalanine, serine, cystine, valine, isoleucine, leucine and aspartic acid than Unsavda patients (P<0.05). Cervical cancer patients with abnormal Savda had low concentration of plasma arginine, but higher concentration of plasma cystine than Unsavda patients (P<0.05). Breast cancer patients with abnormal Savda had higher concentration of plasma leucine, serine, taurine, cystine, tyrosine, valine, isoleucine and asparagine than Unsavda patients (P<0.05). Gastric cancer patients with abnormal Savda had high concentration of plasma cystine but lower concentration of plasma phenylalanine, threonine and arginine than Unsavda patients (P<0.05).@*CONCLUSION@#Different tumor patients with abnormal Savda have common characteristics and significant differences.

Metabolic changes in abnormal savda patients with different types of tumor: a clinical observation / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To explore in vivo metabolic changes in abnormal savda patients with different types of tumor.</p><p><b>METHODS</b>A total of 142 abnormal savda patients with common cancer types were enrolled in this study, and 50 healthy volunteers were recruited as the control group. For each sample, the H Nuclear Magnetic Resonance (NMR) based metabonomic analysis was performed. The free attenuation signal was computed subsection integral. Data obtained were analyzed by the Orthogonal Partial Least-Squares Discriminant Analysis (OPLS-DA).</p><p><b>RESULTS</b>Compared with the control group, leucine, isoleucine, valine, histidine, phenylalanine, tyrosine, alanine, creatine, lactic acid, inositol, alpha-and beta-glucose, unsaturated lipids, very low density lipoprotein (VLDL) significantly decreased (P <0.05), while glycoprotein and carnitine significantly increased (P <0. 05) in the abnormal Savda group.</p><p><b>CONCLUSION</b>Abnormal savda patients with different types of tumor had similar metabonomics changes.</p>

Correlative analysis of neoplasm patients with phlegm-stasis or abnormal Savda syndrome, based on metabonomics.

OBJECTIVE: To investigate plasma samples from neoplasm patients with phlegm-stasis or abnormal Savda syndrome, with NMR spectroscopy, and to analyze their metabolic varieties, characteristics and reciprocity. METHODS: 1H-NMR spectra were analyzed using the orthogonal projection to latent structure with discriminative analysis (OPLS-DA) method with unit variance scaling. The discriminative significance of metabolites was determined by the Pearson's product - moment correlation coefficient. RESULTS: Compared to the control group, neoplasm patients with phlegm - stasis or abnormal Savda syndrome had low concentrations of leucine, isoleucine, valine, alanine, tyrosine, histidine, citrulline, glycoprotein, glutamine, myo-inositol, scyllo-inositol, creatine, alpha-glucose, alpha-glucose and lactate (P < 0.05), and high concentrations of very low density lipoprotein, low density lipoprotein, unsaturated lipid, formate, acetone, acetate, acetoacetate, pyruvate, beta-hydroxybutyrate, carnitine and malonic acid (P < 0.05), with no significant differences between the phlegm - stasis and abnormal Savda syndrome patients. CONCLUSIONS: Neoplasm patients with different syndromes have very similar metabolic changes. A series of abnormalities such as immune dysfunction and oxidative - antioxidative imbalance, occur in neoplasm patients with abnormal Savda or phlegm - stasis syndrome.

Gender related metabonomic analysis of serum and urine samples from Xinjiang healthy Han subjects with magnetic resonance / 中南大学学报(医学版)

OBJECTIVE@#To investigate gender variability in the metabolic serum and urinary profile of healthy Han population in Xinjiang.@*METHODS@#Serum and urinary samples from 92 healthy Han people in Xinjiang were tested by magnetic resonance based metabonomics and pattern recognition analysis performed with orthogonal partial least-squares discriminant analysis (OPLS-DA). The quality of the model was described by parameter R(2)X, R(2)Y, and Q(2).@*RESULTS@#The serum in males had higher levels of very low density lipoprotein, low density lipoprotein, unsaturated lipids, creatinine and acetone than in females, whereas females had higher levels of citrate, choline, glucose and amino acids (including isoleucine, leucine, valine, alanine, citrulline, lysine, methionine, glutamate, phenylalanine, threonine, tyrosine, 1-methyl histidine and glycine) than in males. The urine of males had higher levels of formate, malonic acid, taurine, creatinine than that of females, while females had higher levels of hippurate, γ-aminobutyric acid, succinate, citrate and glutamate than males. The model parameters of serum were R(2)X=0.64, R(2)Y=0.70, and Q(2)=0.67, and those of urine were R(2)X=0.17, R(2)Y=0.70, and Q(2)=0.44.@*CONCLUSION@#The blood and urine from Han population in Xinjiang contain a variety of gender related metabolites, which plays an important role in the research of clinical metabonomics.

Plasma metabonomics analysis of tumor patients of phlegm-stasis syndrome / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To study metabonomic changes in plasma of tumor patients of phlegm-stasis syndrome by Chinese medicine and their in vivo metabolic mechanism.</p><p><b>METHODS</b>1H nuclear magnetic resonance (NMR) based metabonomic analysis was performed on plasma samples from 356 tumor patients of the phlegm-stasis syndrome and 104 tumor patients of the non-phlegm-stasis syndrome, and 50 healthy subjects. The spectrogram integral results were analyzed by orthogonal partial least-squares discriminant analysis (OPLS-DA).</p><p><b>RESULTS</b>Compared with healthy subjects, various amino acids including leucine, alanine, citrulline, tyrosine, histidine, arginine, methionine, isoleucine, valine, acetylcysteine, etc. in the plasma of patients of the phlegm-stasis syndrome were significantly lowered (P <0.05). Glucose, glycoprotein, glutamine, myo-inositol, lactic acid, choline, creatine also significantly decreased (P<0.05). But the plasma formic acid, acetone, acetic acid, acetoacetate, pyruvate, beta-hydroxy butyrate, carnitine, malonic acid, and unsaturated fatty acid, very low density lipoprotein cholesterol (VLDL-C), low density lipoprotein cholesterol (LDL-C) increased in tumor patients of the phlegm-stasis syndrome. Compared with tumor patients of non-phlegm-stasis syndrome, patients of the phlegm-stasis syndrome had obvious lower plasma contents of leucine, alanine, citrulline, tyrosine, histidine, soleucine, valine, glutamine, myo-inositol, scyllo-inositol, lactic acid, creatine (P <0. 05), higher plasma contents of acetone, acetoacetate, unsaturated fatty acid, VLDL-C, alpha-glucose, beta-glucose, glycoprotein, and so on (P <0. 05).</p><p><b>CONCLUSIONS</b>Tumor patients of the phlegm-stasis syndrome had strengthened in vivo fat metabolism and lowered various amino acids. The decreased antioxidation capacities resulted in aggravated cell membrane injuries. The in vivo metabolic disorder was more severe in tumor patients of the phlegm-stasis syndrome than in tumor patients of the non-phlegm-stasis syndrome.</p>

Plasma metabonomic analysis with ¹H nuclear magnetic resonance revealing the relationship of different tumors and the disease homology theory of traditional Uyghur medicine / 中国结合医学杂志

<p><b>OBJECTIVE</b>To investigate the plasma samples obtained from tumor patients using (1)H nuclear magnetic resonance (NMR) spectroscopy, and find the biochemical foundation of abnormal Savda described in traditional Uyghur medicine.</p><p><b>METHODS</b>A total of 170 tumor patients with abnormal Savda syndrome who were confirmed clinically were enrolled in this study, and 50 healthy volunteers were set up as controls. The plasma (1)H NMR spectra were analyzed using the orthogonal projection to latent structure with discriminant analysis (OPLS-DA) method with unit variance scaling. The discriminative significance of the metabolites was determined using the Pearson's product-moment correlation coefficient.</p><p><b>RESULTS</b>Compared with the healthy controls, the tumor patients with abnormal Savda syndrome had uniformly correlative low levels of leucine, isoleucine, valine, histidine, tyrosine, alanine, glutamine, creatine, inositol, α-glucose, and β-glucose (P<0.05), but had significantly high levels of formate, malonic acid, acetone, acetate, acetoacetate, pyruvate, β-hydroxy butyrate, carnitine and lipidtemns such as very low density lipoprotein, low density lipoprotein and unsaturated lipids (P<0.05).</p><p><b>CONCLUSION</b>Tumor patients with abnormal Savda syndrome had similar metabolic changes and characteristics, which indicated a similar pathogenetic process and provides some biochemical basis for traditional Uyghur medicine theory.</p>

[1H-MRS metabonomic analysis of plasma samples of esophageal cancer patients based on different pattern recognition].

OBJECTIVE: To analyze the metabonomic (1)H-MRS of plasma samples from patients with esophageal cancer and healthy controls applying different pattern recognition methods, and to explore the potential of application of (1)H-MR-based metabonomics in clinical research. METHODS: (1)H-MR was performed on plasma samples from 109 EC patients and 50 health controls to analyze the metabonomic variation between EC patients and healthy subjects and the corresponding (1)H-MRS were recorded on Varian Unity ANOVA 600 MHz to perform principal components analysis (PCA), partial least squares discriminant analysis (PLS-DA), orthogonal partial least squares discriminant analysis (OPLS-DA), respectively. RESULTS: OPLS-DA analysis could correctly separate almost all the plasma samples from EC patients and health controls, better than both the PCA and PLS-DA. The plasma levels of leucine, alanine, isoleucine, valine, glycoprotein, lactate, acetone, acetate, choline, isobutyrate, unsaturated lipid, VLDL, LDL, 1-methylhistidine were significantly decreased in EC patients (r total > 0.27, P < 0.05), while that of dimethylamine, α-glucose, ß-glucose, citric acid increased in the EC patients (r total < -0.27, P < 0.05). CONCLUSIONS: The analysis of metabonomic (1)H-MRS of plasma samples by OPLS-DA method can eliminate the influence of non-experimental factors and decrease the heterogeneity of samples. It is useful and of great potential for application in clinical diagnosis and research of esophageal cancer.

[Metabonomic variation of esophageal cancer within different ethnic groups in Xinjiang, China].

OBJECTIVE: To investigate the metabonomic variation between patients with esophageal cancer (EC) and healthy controls, and to analyze the variation between patients with EC. METHODS: H-MR and orthogonal partial least-squares discriminant analysis (OPLS-DA) was performed on 108 plasma specimens from EC patients and 50 health controls, and the metabonomic variation between patients with EC and healthy controls analyzed. RESULTS: OPLS-DA analysis might correctly separate all plasma specimens from health controls and patients with EC, leucine (0.0043 +/- 0.0006, 0.0040 +/- 0.0006), alanine (0.0039 +/- 0.0007, 0.0033 +/- 0.0006), isoleucine (0.0067 +/- 0.0010, 0.0063 +/- 0.0009), valine (0.0037 +/- 0.0005, 0.0035 +/- 0.0006), glycoprotein (0.0123 +/- 0.0043, 0.0102 +/- 0.0022), lactate (0.0342 +/- 0.0113, 0.0258 +/- 0.0085), acetone (0.0027 +/- 0.0023, 0.0017 +/- 0.0008), acetate (0.0007 +/- 0.0001, 0.0006 +/- 0.0001), choline (0.0035 +/- 0.0006, 0.0029 +/- 0.0007), isobutyrate (0.0020 +/- 0.0004, 0.0018 +/- 0.0003), unsaturated lipid (0.0072 +/- 0.0013, 0.0059 +/- 0.0018), VLDL (0.1209 +/- 0.0589, 0.0879 +/- 0.0269), LDL (0.0885 +/- 0.0328, 0.0785 +/- 0.0288), 1-methylhistidine (0.0005 +/- 0.0001, 0.0004 +/- 0.0005) decreased in EC patient' s plasma with statistical significance (r total > 0.27, P < 0.05), dimethylamine (0.0004 +/- 0.0001, 0.0005 +/- 0.0001), alpha-glucose (0.0079 +/- 0.0013, 0.0081 +/- 0.0016), 3-glucose (0.0139 +/- 0.0024, 0.0142 +/- 0.0029), citric acid (0.0044 +/- 0.0008, 0.0106 +/- 0.0058) increase in the EC patient's plasma (r total < -0.27, P < 0.05). There were clear variation between Han and Kazak patients, alanine (0.0031 +/- 0.0005,0.0029 +/- 0.0004), glutamine (0.0010 +/- 0.0001, 0.0009 +/- 0.0001), tyrosine (0.0009 +/- 0.0001, 0.0008 +/- 0.0001), 1-methylhistidine (0.0005 +/- 0.0001, 0.0004 +/- 0.0001) increased in the Han patients (r > 0.35, P> 0.05), carnitine (0.0028 +/- 0.0006) was higher in Kazak patients than in Han patients (0.0025 +/- 0.0004), which had statistical significance (r = - 0.40, P < 0.05). Unsaturated lipid (0.0059 +/- 0.0018, 0.0047 +/- 0.0011), isoleucine (0.0062 +/- 0.0011, 0.0058 +/- 0.0007), alanine (0. 0032 +/- 0.0007, 0.0028 +/- 0.0004), glycoprotein (0.0096 +/- 0.0019, 0.0086 +/- 0.0011), glutamine (0.0011 +/- 0.0001, 0.0009 +/- 0.0001), tyrosine (0.0009 +/- 0.0001, 0.0008 +/- 0.0001), 1-methylhistidine (0.0005 +/- 0.0001, 0.0004 +/- 0.0001) were increased in Uygur patients as compared with Kazak patients, having statistical significance (r > 0.33, P < 0.05), carnitine (0.0027 +/- 0.0005) was higher in Kazak patients than in Uygur patients (0.0025 +/- 0.0004) (r = - 0.36, P < 0.05). CONCLUSION: The results indicate that 1H-MR spectra of plasma analyzed by OPLS-DA statistical methods might completely separate the EC patients from health controls. The metabonomic approach should be helpful in screening of EC patients.