Batur Mamtimin

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.

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.

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.

Plasma metabonomic analysis with 1H nuclear magnetic resonance revealing the relationship of different tumors and the disease homology theory of traditional Uyghur medicine

ObjectiveTo investigate the plasma samples obtained from tumor patients using 1H nuclear magnetic resonance (NMR) spectroscopy, and find the biochemical foundation of abnormal Savda described in traditional Uyghur medicine.MethodsA 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 1H 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.ResultsCompared 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).ConclusionTumor 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.

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.

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.

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.

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 ASMqs 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, β-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, β-glucose, and glutamine concentration increased (P < 0.05). T2DM rats had higher concentrations of α- and β-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 β-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.

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 Students 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.

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.