[Effect of diethylnitrosamine on renal metabolic pathway in rats].

OBJECTIVE: 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.

Metabolomic profiling reveals potential biomarkers in esophageal cancer progression using liquid chromatography-mass spectrometry platform.

Esophageal cancer (EC) is one of the most common malignancies with poor prognosis. Metabolomics has been shown to be a powerful approach to discover the potential biomarkers for cancer diagnosis and prognosis. The goal of this study is to screen potential biomarkers for early diagnosis and prognosis. In this study, 40 tissue samples and the corresponding control samples from the same esophageal squamous cell carcinoma (ESCC) patients were analyzed by liquid chromatography-mass spectrometry (LC-MS)-based metabolomics. 20 potential diagnostic biomarkers were selected. Moreover, 9 metabolites were found to be closely correlated with the pathological feature such as local invasion, lymphatic metastasis and postoperative survival time. Glutamate was correlated with local invasion of tumor, and oleic acid, LysoPC(15:0), uracil, inosine and choline were closely related with the lymphatic metastasis, while glutamine, kynurenine, serine and uracil were related with postoperative survival time. The results indicated that the potential biomarkers discovered by metabolomics could reflect the metabolic characterization of ESCC, and offers a novel approach for early diagnosis, assessment and prognosis of the disease.

Metabonomic signature analysis in plasma samples of glioma patients based on (1)H-nuclear magnetic resonance spectroscopy.

OBJECTIVE: The presence of a glioma is associated with increasing mortality. In this study, nuclear magnetic resonance (NMR) based metabonomics has been applied to investigate the metabolic signatures of a glioma in plasma. The purpose of this study was to assess the diagnostic potential of this approach and gain novel insights into the metabolism of glioma and its systemic effects. METHODS: Plasma samples were collected prospectively by centrifugation of blood samples from patients with a glioma (n = 70) or a control group (n = 70). NMR spectra of these plasma samples were analyzed using orthogonal partial least square discriminant analysis (OPLS-DA) to identify the potential biomarkers. RESULTS: The OPLS-DA model showed a good differentiation between the glioma and the control groups. A total of 20 metabolites were identified, which are closely correlating with the presence of a glioma. Compared to the control group, patients with a glioma were associated with lower concentrations of isoleucine, leucine, valine, lactate, alanine, glycoprotein, glutamate, citrate, creatine, myo-inositol, choline, tyrosine, phenylalanine, 1-methylhistidine, α-glucose, ß-glucose, and higher concentrations of very low density lipoprotein, low density lipoprotein (LDL), unsaturated lipid, and pyruvate. These 20 metabolites, which are involved in energy, fatty acid, and amino acid metabolism, may be associated with a human glioma. CONCLUSION: Our study is the first one to identify the plasma metabolites that have the potential to distinguish between patients with a glioma and healthy subjects. NMR-based metabonomics provides a good sensitivity and selectivity in differentiating the healthy control group from patients suffering form the disease. Plasma metabolic profiling may have a potential in diagnosing a glioma in the early phase and may help in enhancing our understanding of its underlying mechanisms.

An magnetic resonance-based plasma metabonomic investigation on abnormal Savda in different complicated diseases.

OBJECTIVE: To provide potential evidence for the existence of abnormal Savda, we assessed host metabonomic responses and dynamic changes occurring in various diseases using 1H nuclear magnetic resonance (NMR)-based metabonomics. METHODS: Plasma samples taken from patients with complicated diseases with abnormal Savda (n = 140, including 35 cases each of diabetes, asthma, breast cancer, and cervical carcinoma) and from healthy controls (n = 35) were analyzed by 1H NMR (600 MHz), and the spectral profiles were analyzed by multivariate analysis using orthogonal projection to latent structure with discriminant analysis. RESULTS: Supervised modeling of the data provided very good discrimination between patients and healthy controls. Compared with the healthy controls, the patient groups with different disease conditions displayed similar metabolic changes, characterized by lower creatine, creatinine, lactate, and amino acid levels (including isoleucine, leucine, valine, alanine, and 1-methylhistidine) and higher lipid levels (very low-density lipoproteins and unsaturated lipids). Additionally, cancer patients (breast and cervical) showed decreased myo-inositol, a-glucose, and beta-glucose, and increased pyruvate and carnitine in plasma. CONCLUSION: The data indicate that decreased oxidative defense, liver function abnormalities, amino acid deficiencies, and energy metabolism disorders are common characteristics of complicated diseases, which may be related to the formation of abnormal Savda.

Plasma amino acid profiling of cancer patients with abnormal savda based on high-performance liquid chromatography.

OBJECTIVE: To investigate metabolic signatures in plasma of cancer patients with abnormal Savda using plasma-free amino acid profiles, and to evaluate the diagnostic potential of these profiles for the detection and explanation of the mechanisms of different symptoms in traditional Uyghur medicine. METHODS: Plasma samples from cancer patients with abnormal Savda (n = 85) or non-abnormal Savda (n = 105) and a healthy control group (n = 65) were analyzed using high-performance liquid chromatography (HPLC). Orthogonal projection to latent structures with discriminant analysis was used for the classification and prediction of abnormal Savda, and spectral profiles were subjected to Student's t-tests to assess statistical significance. RESULTS: Compared with the healthy group, the levels of aspartic acid, glutamate, glycine, histidine, arginine, threonine, alanine, proline, methionine, isoleucine, leucine and phenylalanine decreased significantly in plasma of cancer patients with abnormal Savda (all P < 0.05). Serine, cystine, tyrosine, valine and lysine levels showed no significant differences (all P > 0.05). Compared with non-abnormal Savda syndrome patients, abnormal Savda syndrome patients showed high concentrations of glutamate, serine, valine, isoleucine, leucine and phenylalanine (all P < 0.05). The remaining plasma amino acids showed no significant differences (all P > 0.05). CONCLUSION: Plasma-free amino acid profiling has the potential to assist in understanding and determining abnormal Savda. A HPLC-based metabonomic platform could be a powerful tool for the classification of symptoms in traditional medicine.

[Metabolic changes in abnormal savda patients with different types of tumor: a clinical observation].

OBJECTIVE: To explore in vivo metabolic changes in abnormal savda patients with different types of tumor. METHODS: 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). RESULTS: 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. CONCLUSION: Abnormal savda patients with different types of tumor had similar metabonomics changes.

[Plasma amino acid profiling of "same symptom for different disease" in Uyghur medicine based on high-performance liquid chromatography].

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.

Plasma-free amino acid profiling of cervical cancer and cervical intraepithelial neoplasia patients and its application for early detection.

In this study, plasma-free amino acid profiles were used to investigate pre-cancerous cervical intraepithelial neoplasia (CIN) and cervical squamous cell carcinoma (CSCC) metabolic signatures in plasma. Additionally, the diagnostic potential of these profiles was assessed, as well as their ability to provide novel insight into CSCC metabolism and systemic effects. Plasma samples from CIN patients (n = 26), CSCC patients (n = 22), and a control healthy group (n = 35) were analyzed by high-performance liquid chromatography, and their spectral profiles were subjected to the t test for statistical significance. Potential metabolic biomarkers were identified using database comparisons that examine the significance of metabolites. Compared with healthy controls, patients with CIN and CSCC demonstrated lower levels of plasma amino acids; plasma levels of arginine and threonine were increased in CIN patients but were decreased in cervical cancer patients. Additionally, the levels of a larger group of amino acids (aspartate, glutamate, asparagine, serine, glycine, histidine, taurine, tyrosine, valine, methionine, lysine, isoleucine, leucine, and phenylalanine) were gradually reduced from CIN to invasive cancer. These findings suggest that plasma-free amino acid profiling has great potential for improving cancer screening and diagnosis and for understanding disease pathogenesis. Plasma-free amino acid profiles may have the potential be used to determine cancer diagnoses in the early stage from a single blood sample and may enhance our understanding of its mechanisms.

Revealing the metabonomic variation of EC using ¹H-NMR spectroscopy and its association with the clinicopathological characteristics.

In this study, (1)H NMR-based metabonomics has been applied to investigate esophageal cancer metabolic signatures in plasma and urine, purpose of assessing the diagnostic potential of this approach and gaining novel insights into esophageal cancer metabolism and systemic effects. Plasma and urine samples from esophageal cancer patients (n = 108) and a control healthy group (n = 40) were analyzed by Nuclear Magnetic Resonance (NMR) spectroscopy (600 MHz), and their spectral profiles subjected to Orthogonal Projections to Latent Structures (OPLS-DA) for multivariate statistics. Potential metabolic biomarkers were identified using data base comparisons used for examining the significance of metabolites. Compared to healthy controls, esophageal cancer plasma had higher levels of dimethylamine, α-glucose, ß-glucose, citric acid, together with lower levels of Leucine, alanine, isoleucine, valine, glycoprotein, lactate, acetone, acetate, choline, isobutyrate, unsaturated lipid, VLDL, LDL, 1-methylhistidine; Compared to healthy controls, esophageal cancer urine had higher levels of Mannitol, glutamate, γ-propalanine, phenylalanine, acetate, allantoin, pyruvate, tyrosine, ß-glucose and guinolinate, together with lower levels of N-acetylcysteine, valine, dihydrothymine, hippurate, methylguanidine, 1-methylnicotin- amide and Citric acid; Very good discrimination between cancer and control groups was achieved by multivariate modeling of plasma and urinary profiles. (1)H NMR-based metabolite profiling analysis was shown to be an effective approach to differentiating between patients with EC and healthy subjects. Good sensitivity and selectivity were shown by using the metabolite markers discovered to predict the classification of samples from the healthy control group and the patients with the disease. Plasma and urine metabolic profiling may have potential for early diagnosis of EC and may enhance our understanding of its mechanisms.

[Gender related metabonomic analysis of serum and urine samples from Xinjiang healthy Han subjects with magnetic resonance].

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.

Receptor for activated C kinase 1 promotes cervical cancer lymph node metastasis via the glycolysis­dependent AKT/mTOR signaling.

Cervical cancer (CC), an aggressive form of squamous cell carcinoma, is characterized by early­stage lymph node metastasis and an extremely poor prognosis. The authors have previously demonstrated that patients with CC have aberrant glycolysis. The upregulation of receptor for activated C kinase 1 (RACK1) is associated with CC lymph node metastasis (LNM). However, its role in mediating aerobic glycolysis in CC LNM remains unclear. In the present study, 1H nuclear magnetic resonance analysis revealed a significant association between RACK1 expression and the glycolysis/gluconeogenesis pathway. Additionally, RACK1 knockdown inhibited aerobic glycolysis and lymphangiogenesis in vitro and suppressed CC LNM in vivo. Furthermore, protein kinase B (AKT)/mammalian target of rapamycin (mTOR) signaling was identified as a critical RACK1­regulated pathway that increased lymphangiogenesis in CC. Co­immunoprecipitation, immunofluorescence and western blot analysis revealed that RACK1 activated AKT/mTOR signaling by interacting with insulin­like growth factor 1 receptor (IGF1R). POU class 2 homeobox 2 (POU2F2) bound to the RACK1 promoter and regulated its transcription, thereby functionally contributing to glycolysis and lymphangiogenesis in CC. Of note, the administration of 2­deoxy­D­glucose, which attenuates glycolysis, inhibited RACK1­induced lymphangiogenesis in CC. The correlations between RACK1, IGF1R, POU2F2 and hexokinase 2 were further confirmed in CC tissues. Thus, RACK1 plays a crucial role in CC tumor LNM by regulating glycolysis via IGF1R/AKT/mTOR signaling. Thus, the targeting of the POU2F2/RACK1/IGF1R/AKT/mTOR signaling pathway may provide a novel treatment strategy for CC.

[Plasma metabonomics analysis of tumor patients of phlegm-stasis syndrome].

OBJECTIVE: To study metabonomic changes in plasma of tumor patients of phlegm-stasis syndrome by Chinese medicine and their in vivo metabolic mechanism. METHODS: 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). RESULTS: 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). CONCLUSIONS: 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.

Metabonomic analysis of rat urine ¹H magnetic resonance spectra based on different normalization methods.

OBJECTIVE: To study metabonomics in the urine of rats of different genders by magnetic resonance (MR) with 2 normalization methods. METHODS: Different normalization methods such as mean-centering not scaling (Ctr) and unit variance scaling (UV) were used before orthogonal to partial least squares discriminant analysis(OPLS-DA). Distinguished metabolites in the urine of different gender rats were analyzed by calculating the correlation coefficients. RESULTS: The data normalized by Ctr before OPLS-DA analysis revealed high degree conception metabolites in the urine such as valine, alanine, acetate, ornithine, aminohippurate, phenylethylamine, cytosine, citrate, dimethylamine, allantoin, methylamine, fumarate and one unknown metabolite whose chemical shift was δ4.14. Data normalized by UV before OPLS-DA analysis revealed the above 12 high degree conception metabolites except citrate, and also low degree conception metabolites such as thiamine, creatinine, formate and one unknown metabolite whose chemical shift was δ2.92. CONCLUSION: Unit variance scaling is a more effective normalization method in metabonomic analysis.

Differential integrative omic analysis for mechanism insights and biomarker discovery of abnormal Savda syndrome and its unique Munziq prescription.

Research has shown that many cancers have acommon pathophysiological origin and often present with similar symptoms. In terms of Traditional Uighur Medicine (TUM) Hilit (body fluid) theory, abnormal Savda syndrome (ASS) formed by abnormal Hilit is the common phenotype of complex diseases and in particular tumours. Abnormal Savda Munziq (ASMq), one representative of TUM, has been effective in the treatment of cancer since ancient times. Despite the physiopathology of ASS, the relationship between causative factors and the molecular mechanism of ASMq are not fully understood. The current study expanded upon earlier work by integrating traditional diagnostic approaches with others utilizing systems biology technology for the analysis of proteomic (iTRAQ) and metabolomic ((1)H-NMR) profiles of Uighur Medicine target organ lesion (liver) tumours. The candidate proteins were analyzed by enrichment analysis of the biological process and biomarker filters. Subsequently, 3Omics web-based tools were used to determine the relationships between proteins and appropriate metabolites. ELISA assay and IHC methods were used to verify the proteomic result; the protein von Willebrand factor (vWF) may be the "therapeutic window" of ASMq and biomarkers of ASS. This study is likely to be of great significance for the standardization and modernization of TUM.

Metabolic differentiation and classification of abnormal Savda Munziq's pharmacodynamic role on rat models with different diseases by nuclear magnetic resonance-based metabonomics.

BACKGROUND: Abnormal Savda Munziq (ASMq) is a traditional Uyghur herbal preparation used as a therapy for abnormal Savda-related diseases. In this study, we investigate ASMq's dynamic effects on abnormal Savda rat models under different disease conditions. MATERIALS AND METHODS: Abnormal Savda rat models with hepatocellular carcinoma (HCC), type 2 diabetes mellitus (T2DM), and asthma dosed of ASMq. Serum samples of each animal tested by nuclear magnetic resonance spectroscopy and analyzed by orthogonal projection to latent structure with discriminant analysis. RESULTS: Compared with healthy controls, HCC rats had higher concentrations of amino acids, fat-related metabolites, lactate, myoinositol, and citrate, but lower concentrations of α-glucose, ß-glucose, and glutamine. Following ASMq treatment, the serum acetone very low-density lipoprotein (VLDL), LDL, unsaturated lipids, acetylcysteine, and pyruvate concentration decreased, but α-glucose, ß-glucose, and glutamine concentration increased (P < 0.05). T2DM rats had higher concentrations of α- and ß-glucose, but lower concentrations of isoleucine, leucine, valine, glutamine, glycoprotein, lactate, tyrosine, creatine, alanine, carnitine, and phenylalanine. After ASMq treated T2DM groups showed reduced α- and ß-glucose and increased creatine levels (P < 0.05). Asthma rats had higher acetate, carnitine, formate, and phenylalanine levels, but lower concentrations of glutamine, glycoprotein, lactate, VLDL, LDL, and unsaturated lipids. ASMq treatment showed increased glutamine and reduced carnitine, glycoprotein, formate, and phenylalanine levels (P < 0.05). CONCLUSION: Low immune function, decreased oxidative defense, liver function abnormalities, amino acid deficiencies, and energy metabolism disorders are common characteristics of abnormal Savda-related diseases. ASMq may improve the abnormal metabolism and immune function of rat models with different diseases combined abnormal Savda.

Metabolomic changes in patients with chronic obstructive pulmonary disease with abnormal Savda syndrome.

The aim of this study was to determine the metabolic biomarkers for abnormal Savda syndrome in patients with chronic obstructive pulmonary disease (COPD). Based on Traditional Uyghur Medicine (TUM) theory, a total of 103 patients with COPD were classified into abnormal Savda and non-abnormal Savda syndrome groups and 52 healthy volunteers acted as the control group. Blood samples from the three groups were analyzed using nuclear magnetic resonance (NMR) spectroscopy combined with orthogonal projection to latent structure-discriminant analysis. NMR tests showed that the regional distributions of the patients with COPD with abnormal Savda syndrome, those with non-abnormal Savda syndrome and the control group were completely separate (P>0.05). The patients with COPD with abnormal Savda syndrome exhibited relatively low levels of amino acids, glycoproteins and unsaturated lipids (P<0.05) but significantly higher levels of lactic acid, carnitine, acetone and acetoacetate (P<0.05) compared with the healthy controls. Abnormal Savda syndrome was one of the main types of syndrome among the patients with COPD; increased age, a longer duration of illness and a higher disease severity were characteristic of this type of syndrome. In addition, the present study provided biochemical evidence for the TUM theory-based classification of patients with COPD; these biomarkers can be used in the clinic for the diagnosis of COPD with abnormal Savda syndrome. The study also demonstrated that the plasma metabolic disorder in patients with COPD with abnormal Savda syndrome was more serious than that in the control and COPD with non-abnormal Savda syndrome groups. The plasma metabolic disorder was also associated with a low immune function of the body and endocrine and energy metabolism disorders.

Plasma free amino acid profiling of esophageal cancer using high-performance liquid chromatography spectroscopy.

AIM: To perform plasma free amino acid (PFAA) profiling of esophageal squamous cell carcinoma (ESCC) patients at different pathological stages and healthy subjects. METHODS: Plasma samples from ESCC patients (n = 51) and healthy control adults (n = 60) were analyzed by high-performance liquid chromatography (HPLC). The ESCC patients included moderate/poorly-differentiation (n = 24), lymph node metastasis (n = 17) and clinical stage > Ib2 (n = 36). Partial least squares discriminant analysis was performed to demonstrate that the PFAA metabolic patterns enabled discrimination between ESCC patients and controls, and the Student t test was applied to assess significant differences in PFAA concentrations between the two groups. RESULTS: There were significant differences in the PFAA profiles between controls and ESCC patients. Compared with healthy controls, the levels of Asp, Glu, Gly, His, Thr, Tau, Ala, Met, Ile, Leu, and Phe were decreased in ESCC patients, but Cys was increased. There exists a strong correlation between PFAA profiles and clinicopathological characteristics in ESCC patients. The levels of many PFAAs (i.e., Glu, Asp, Ser, Gly, Tau, Ala, Tyr, Val, Ile, and Leu) were related to pathological grading, lymph node metastasis, and ESCC clinical stage. Very good discrimination between ESCC patients and control subjects was achieved by multivariate modeling of plasma profiles. CONCLUSION: HPLC-based plasma profiling analysis was shown to be an effective approach to differentiate between ESCC patients and controls. PFAA profiles may have potential value for screening or diagnosing ESCC.

Association of the plasma and tissue riboflavin levels with C20orf54 expression in cervical lesions and its relationship to HPV16 infection.

Riboflavin deficiency can cause a variety of metabolic problems that lead to skin and mucosal disorders. Limited evidence suggests that high intake of riboflavin may reduce overall risks of cancer. However, association of this deficiency with cervical cancer and precancerous lesions are still not definitively known. In this study, we characterized the relationship between plasma and tissue riboflavin levels and C20orf54 protein expression in patients with cervical intraepithelial neoplasia (CIN) and cervical squamous cell carcinoma (CSCC) as well as the relationship of these levels with human papillomavirus virus 16, 18 (HPV16/18) infections. High-performance liquid chromatography (HPLC) was used to measure blood riboflavin levels in patients with CIN and CSCC, and an enzyme-linked immunosorbent assay (ELISA) was used to determine tissue riboflavin levels in patients with CSCC and matched normal mucous epithelia. The expression of C20orf54 in fresh CSCC and matched tissues were detected by qRT-PCR and western blot, respectively. And it was further confirmed by immunohistochemistry (IHC) with formalin-fixed, paraffin-embedded CIN and CSCC. An HPV genotyping chip was used to analyze HPV infection and typing. The results showed that patients with CIN and CSCC had decreased plasma riboflavin levels as compared with normal controls. There was also significantly decreased riboflavin in tissues from CSCC patients, when compared with normal cervical epithelia. C20orf54 expression were significantly up-regulated in CSCC compared to matched control on both mRNA and protein level. Tissue riboflavin levels were significantly lower in HPV16/18 positive tissue compared with HPV16/18-negative tissue, and an inverse association was found between tissue riboflavin levels and C20orf54 mRNA and protein expression in CSCC. Additionally, C20orf54 was significantly correlated with tumor stages. In conclusion, C20orf54 tend to play a protective role in Uyghur cervical carcinogenesis of which modulating riboflavin absorption, and it is also related with HPV infection.

Metabonomic signature analysis of cervical carcinoma and precancerous lesions in women by (1)H NMR spectroscopy.

(1)H nuclear magnetic resonance (NMR)-based metabonomics has been used to characterize the metabolic profiles of cervical intraepithelial neoplasia (CIN) and cervical squamous cell carcinoma (CSCC). Principal component analysis (PCA) and orthogonal partial least-squares discriminant analysis (OPLS-DA) were used to model the systematic variation related to patients with CIN or CSCC with healthy controls. Potential metabolic biomarkers were identified using database comparisons, and the one-way analysis of variance (ANOVA) test was used to examine the significance of the metabolites. Compared with plasma obtained from the healthy controls, plasma from patients with CIN had higher levels of very-low density lipoprotein (VLDL), acetone, unsaturated lipid and carnitine, together with lower levels of creatine, lactate, isoleucine, leucine, valine, alanine, glutamine, histidine, glycine, acetylcysteine, myo-inositol, choline and glycoprotein. Plasma from patients with CSCC had higher levels of acetate and formate, together with lower levels of creatine, lactate, isoleucine, leucine, valine, alanine, glutamine, histidine and tyrosine compared with the plasma of the healthy controls. In addition, compared with the plasma of patients with CIN, the plasma of CSCC patients had higher levels of acetate, formate, lactate, isoleucine, leucine, valine, alanine, glutamine, histidine, tyrosine, acetylcysteine, myo-inositol, glycoprotein, α-glucose and ß-glucose, together with lower levels of acetone, unsaturated lipid and carnitine. Moreover, the profiles showed high feasibility and specificity by statistical analysis with OPLS-DA compared to the Thinprep cytology test (TCT) by setting the histopathological outcome as standard. The metabolic profile obtained for cervical cancer is significant, even for the precancerous disease. This suggests a systemic metabolic response to cancer, which may be used to identify potential early diagnostic biomarkers of the cancer and to establish clinical diagnostic methods.

Decreased blood riboflavin levels are correlated with defective expression of RFT2 gene in gastric cancer.

AIM: To investigate the relationship between blood riboflavin levels and riboflavin transporter 2 (RFT2) gene expression in gastric carcinoma (GC) development. METHODS: High-performance liquid chromatography was used to detect blood riboflavin levels in patients with GC. Real-time fluorogenic quantitative polymerase chain reaction and immunohistochemistry were used to analyze the expression of RFT2 mRNA and protein in samples from 60 GC patients consisting of both tumor and normal tissue. RESULTS: A significant decrease in the RFT2 mRNA levels was detected in GC samples compared with those in the normal mucous membrane (0.398 ± 0.149 vs 1.479 ± 0.587; P = 0.040). Tumors exhibited low RFT2 protein expression (75%, 16.7%, 8.3% and 0% for no RFT2 staining, weak staining, medium staining and strong staining, respectively), which was significantly lower than that in the normal mucous membrane (10%, 16.7%, 26.7% and 46.7% for no RFT2 staining, weak staining, medium staining and strong staining, respectively; P < 0.05). Tumors with low RFT2 expression were significantly associated with tumor stage and histological grade. Moreover, a significantly decrease in Uyghur patients was observed compared with Han patients. However, other parameters-gender, tumor location and lymph node metastasis-showed no significant relationship with RFT2 expression. Blood riboflavin levels were reverse correlated with development of GC (1.2000 ± 0.97569 ng/mL in high tumor stage patients vs 2.5980 ± 1.31129 ng/mL in low tumor stage patients; P < 0.05). A positive correlation of plasma riboflavin levels with defective expression of RFT2 protein was found in GC patients (χ² = 2.619; P = 0.019). CONCLUSION: Defective expression of RFT2 is associated with the development of GC and this may represent a mechanism underlying the decreased plasma riboflavin levels in GC.