This study investigated the potential mechanism of Cordyceps militaris(CM) against non-small cell lung cancer(NSCLC) based on serum untargeted metabolomics. Specifically, Balb/c nude mice were used to generate the human lung cancer A549 xenograft mouse model. The tumor volume, tumor weight, and tumor inhibition rate in mice in the model, cisplatin, Cordyceps(low-, medium-, and high-dose), and CM(low-, medium-, and high-dose) groups were compared to evaluate the influence of CM on lung cancer. Gas chromatography-mass spectrometry(GC-MS) was used for the analysis of mouse serum, SIMCA 13.0 for the compa-rison of metabolic profiles, and MetaboAnalyst 5.0 for the analysis of metabolic pathways. According to the pharmacodynamic data, the tumor volume and tumor weight of mice in high-dose CM group and cisplatin group decreased as compared with those in the model group(P<0.05 or P<0.01). The results of serum metabolomics showed that the metabolic profiles of the model group were significantly different from those of the high-dose CM group, and the content of endogenous metabolites was adjusted to different degrees. A total of 42 differential metabolites and 7 differential metabolic pathways were identified. In conclusion, CM could significantly inhibit the tumor growth of lung cancer xenograft mice. The mechanism is the likelihood that it influences the aminoacyl-tRNA biosynthesis, the metabolism of D-glutamine and D-glutamate, metabolism of alanine, aspartate, and glutamate, metabolism of glyoxylate and dicarboxylic acid, biosynthesis of phenylalanine, tyrosine, and tryptophan, arginine biosynthesis as well as nitrogen metabolism. This study elucidated the underlying mechanism of CM against NSCLC from the point of metabolites. The results would lay a foundation for the anticancer research and clinical application of CM.
Alanine/metabolism* ; Animals ; Arginine/metabolism* ; Aspartic Acid ; Carcinoma, Non-Small-Cell Lung/drug therapy* ; Cisplatin/pharmacology* ; Cordyceps ; Glutamic Acid ; Glutamine ; Glyoxylates/metabolism* ; Humans ; Lung Neoplasms/drug therapy* ; Metabolomics/methods* ; Mice ; Mice, Nude ; Nitrogen/metabolism* ; Phenylalanine/metabolism* ; RNA, Transfer/metabolism* ; Tryptophan/metabolism* ; Tyrosine/metabolism*

OBJECTIVE: To compare the analgesic effect of Jin Ling Zi Powder (JLZ) and its two single herbs. METHODS: The hot plate method was used to induce pain. Totally 36 mice were randomly divided into 6 groups by a complete random design, including control, model, aspirin (ASP, 0.14 g/kg body weight), JLZ (14 g/kg body weight), Corydalis yanhusuo (YHS, 14 g/kg body weight), and Toosendan Fructus (TF, 14 g/kg body weight) groups, 6 mice in each group. The mice in the control and model groups were given the same volume of saline, daily for 2 consecutive weeks. At 30, 60, 90, and 120 min after the last administration, the pain threshold of mice in each group was measured, and the improvement rate of pain threshold was calculated. Serum endogenous metabolites were analyzed by gas chromatography-mass spectrometry (GC-MS). RESULTS: There was no statistical difference in pain threshold among groups before administration (P>0.05). After 2 weeks of administration, compared with the model group, the pain threshold in JLZ, YHS, TF and ASP groups were increased to varying degrees (P<0.05). JLZ had the best analgesic effect and was superior to YHS and TF groups. A total of 14 potential biomarkers were screened in serum data analysis and potential biomarkers levels were all reversed to different degrees after the treatment with JLZ and its single herbs. These potential biomarkers were mainly related to glyoxylate and dicarboxylate metabolism, glycine, serine and threonine metabolism, valine, leucine and isoleucine biosynthesis, aminoacyl-tRNA biosynthesis and inositol phosphate metabolism. CONCLUSIONS The analgesic mechanism of JLZ and YHS was mainly due to the combination of glycine and its receptor, producing post-synaptic potential, reducing the excitability of neurons, and weakening the afferent effect of painful information.
Animals ; Mice ; Analgesics/therapeutic use* ; Aspirin/pharmacology* ; Biomarkers ; Body Weight ; Drugs, Chinese Herbal/therapeutic use* ; Glycine ; Glyoxylates ; Inositol Phosphates ; Isoleucine ; Leucine ; Metabolomics/methods* ; Powders ; RNA, Transfer ; Serine ; Threonine ; Valine

OBJECTIVETo explore the intergrating of N-isopropyl oxamate and serum protein and establish a high performance liquid chromatography (HPLC) detection method of N-isopropyl oxamate (specific inhibitor of testis-specific lactate dehydrogenase (LDH-C4)) in the blood of plateau pikas.

METHODSTwenty highland pika 150-200 g, were randomly divided into two groups (n = 10): control group and inhibitor group. Different concentrations of N-isopropyl oxamate were added to examine the intergrating of N-isopropyl oxamate and serum protein. In order to determine its concentration in the pika blood accurately, we used the method of adding trypsin to incubate the serum first, followed by trichloroacetic acid treatment and detecting by HPLC. Results: When the concentrations of N-isopropyl oxamate in the pika serum were added to 0.05 mmol/L, 0.1 mmol/L, 1 mmol/L, 10 mmol/L, 16.7 mmol/L, 33.3 mmol/L and 100 mmol/L, the intergrating rates between N-isopropyl oxamate and plateau pika serum were 100%, 100%, 100%, 86.84%, 54.11%, 40.10% and 20.18%, respectively. The method established in this paper was good on recovery rates, precision and stability. A good linearity was obtained in the range of 0.0125-0.25 mmol/L. When the concentrations of N-isopropyl oxamate in the serum were added to 0. 15 mmol/L,0.3 mmol/L and 1 mmol/L, the recovery rates were 98.05%, 98.98% and 98.12%, respectively; the precision relative standard deviation( RSD) of concentrations were 1.17%, 0.92% and 0.83%, respectively; the stability relative standard deviation (RSD) of concentrations were 1.38%, 1.40% and 0.88%, respectively. The repeatability RSD of the method was 1.76%. Quantitative limit was 0.0125 mmol/L.

CONCLUSIONN-isopropyl oxamate has a strong affinity with plateau pika serum protein that can't be accurately determined with common HIPLC method. It can be accurately determined in the blood by adding trypsinto digest the serum protein first, followed by adding trichloroacetic acid to precipitate the protein.

Animals ; Chromatography, High Pressure Liquid ; methods ; Lagomorpha ; Male ; Oxamic Acid ; analogs & derivatives ; blood

The plateau pika (Ochotona curzoniae) has a strong adaptability to hypoxic plateau environment. We found that the sperm-specific lactate dehydrogenase (LDH-C4) gene Ldh-c expressed in plateau pika cardiac muscle. In order to shed light on the effect of LDH-C4 on the anaerobic glycolysis in plateau pika cardiac muscle, 20 pikas were randomly divided into the inhibitor group and the control group, and the sample size of each group was 10. The pikas of inhibitor group were injected with 1 mL 1 mol/L N-isopropyl oxamate, a specific LDH-C4 inhibitor, in biceps femoris muscle of hind legs, each leg with 500 μL. The pikas of control group were injected with the same volume of normal saline (0.9% NaCl). The mRNA and protein expression levels of Ldh-c gene in plateau pika cardiac muscle were determined by real-time PCR and Western blot. The activities of LDH, and the contents of lactate (LD) and ATP in cardiac muscle were compared between the inhibitor group and the control group. The results showed that 1) the expression levels of Ldh-c mRNA and protein were 0.47 ± 0.06 and 0.68 ± 0.08, respectively; 2) 30 min after injection of 1 mL 1 mol/L N-isopropyl oxamate in biceps femoris muscle, the concentration of N-isopropyl oxamate in blood was 0.08 mmol/L; 3) in cardiac muscle of the inhibitor group and the control group, the LDH activities were (6.18 ± 0.48) U/mg and (9.08 ± 0.58) U/mg, the contents of LD were (0.21 ± 0.03) mmol/g and (0.26 ± 0.04) mmol/g, and the contents of ATP were (4.40 ± 0.69) nmol/mg and (6.18 ± 0.73) nmol/mg (P < 0.01); 5) the inhibition rates of N-isopropyl oxamate to LDH, LD and ATP were 31.98%, 20.90% and 28.70%, respectively. The results suggest that Ldh-c expresses in cardiac muscle of plateau pika, and the pika cardiac muscle may get at least 28% ATP for its activities by LDH-C4 catalyzed anaerobic glycolysis, which reduces the dependence on oxygen and enhances the adaptation to the hypoxic environments.
Acclimatization ; Animals ; Glycolysis ; Hypoxia ; Isoenzymes ; genetics ; metabolism ; L-Lactate Dehydrogenase ; genetics ; metabolism ; Lactic Acid ; analysis ; Lagomorpha ; genetics ; Male ; Myocardium ; enzymology ; Oxamic Acid ; analogs & derivatives ; Oxygen ; RNA, Messenger

OBJECTIVETo develop a method for determination of mandelic acid (MA) and phenylglyoxylic acid (PGA) in urine by reagent-free ion chromatography.

METHODSIon chromatography was performed on an AS19 column with a gradient elution solution containing 10-35 mmoL/L KOH at a flow rate of 1.00 ml/min, and MA and PGA were detected at ultraviolet wavelengths of 225 nm and 254 nm, respectively. The samples were diluted 10 times with purified water, then purified on a silver column to remove high concentrations of chloride ion, and injected after being filtered through a 0.2-µm m filter membrane.

RESULTSThe recoveries of standard addition of MA and PGA were 96.5% and 99.3%, respectively, with both relative standard deviations less than 5.0%. Good linear relationships were noted in the range of 1.0-100.0 mg/L for both MA and PGA (r >0.9995). The detection limits of MA and PGA were 0.02 mg/L and 0.05 mg/L, respectively; the minimum detectable concentrations of MA and PGA were 0.2 mg/L and 0.5 mg/L (when the sampling amount was 5.0 ml and diluted to 50.0 ml with water, and the injection volume was 300 µL).

CONCLUSIONSThis method is fast, convenient, and highly sensitive and selective. It can be used for the analysis of MA and PGA in the urine of styrene-exposed workers.

Chromatography, Ion Exchange ; Glyoxylates ; urine ; Humans ; Mandelic Acids ; urine ; Styrene

OBJECTIVETo investigate the role of genetic polymorphisms of epoxide hydrolase 1 (EPHX1) in the metabolism of styrene in vivo.

METHODSFifty-six styrene-exposed workers, who worked in the painting workshop of an enterprise for manufacturing glass fiber-reinforced plastic yachts in Shandong Province, China for over one year and were protected in approximately the same way, were selected as study subjects. The 8-hour time-weighted average concentration (8 h-TWA) of styrene and the concentrations of mandelic acid (MA) and phenyl glyoxylic acid (PGA) as urinary metabolites were measured. The genetic polymorphisms of EPHX1 were detected by polymerase chain reaction-restriction fragment length polymorphism analysis.

RESULTSThe urinary concentrations of MA and PGA were 177.25±82.36 mg/g Cr and 145.91±69.73 mg/g Cr, respectively, and the 8 h-TWA of styrene was 133.28±95.81 mg/m3. Urinary concentrations of MA and PGA were positively correlated with 8 h-TWA of styrene (R=0.861, P < 0.05; R=0.868, P < 0.05). The subjects were divided into high-exposure group (8 h-TWA >50 mg/m(3)) and low-exposure group (8 h-TWA ≤ 50 mg/m(3), and in the two groups, the urinary concentrations of MA and PGA were significantly higher in the individuals carrying high-activity genotypes of EPHX1 than in those carrying low-activity genotypes of EPHX1 (P < 0.05).

CONCLUSIONGenetic polymorphisms of EPHX1 play an important role in the metabolic process of styrene in vivo.

Adult ; Air Pollutants, Occupational ; pharmacokinetics ; China ; Epoxide Hydrolases ; genetics ; Glyoxylates ; urine ; Humans ; Male ; Mandelic Acids ; urine ; Occupational Exposure ; Polymorphism, Genetic ; Styrene ; pharmacokinetics

Ethylene glycol (EG) is a sweet-tasting, odorless organic solvent found in many agents, such as anti-freeze. EG is composed of four organic acids: glycoaldehyde, glycolic acid, glyoxylic acid and oxalic acid in vivo. These metabolites are cellular toxins that can cause cardio-pulmonary failure, life-threatening metabolic acidosis, central nervous system depression, and kidney injury. Oxalic acid is the end product of EG, which can precipitate to crystals of calcium oxalate monohydrate in the tubular lumen and has been linked to acute kidney injury. We report a case of EG-induced oxalate nephropathy, with the diagnosis confirmed by kidney biopsy, which showed acute tubular injury of the kidneys with extensive intracellular and intraluminal calcium oxalate monohydrate crystal depositions.
Acidosis ; Acute Kidney Injury ; Biopsy ; Calcium Oxalate ; Central Nervous System ; Depression ; Ethylene Glycol ; Ethylenes ; Glycolates ; Glyoxylates ; Kidney ; Oxalic Acid

Primary hyperoxaluria is a rare disorder of glyoxylate metabolism in which hepatic enzyme deficiencies result in overproduction of oxalate. The resulting elevation of urinary oxalate excretion leads to recurrent urolithiasis and progressive nephrocalcinosis. End-stage renal disease frequently occurs and is accompanied by systemic oxalate deposition along with its harmful effects. With the rarity and various clinical heterogeneity of the disease, the high proportion of patients in whom diagnosis is made after advanced renal failure have developed it. On account of its high rate of graft loss associated with primary hyperoxaluria, isolated kidney transplantation has been replaced by combined liver/kidney transplantation. In this report, we describe a case of primary hyperoxaluria with kidney graft failure who had a history of recurrent renal stones.
Glyoxylates ; Humans ; Hyperoxaluria, Primary ; Kidney ; Kidney Failure, Chronic ; Kidney Transplantation ; Nephrocalcinosis ; Population Characteristics ; Renal Insufficiency ; Transplantation, Homologous ; Transplants ; Urolithiasis

OBJECTIVES: This study was designed to investigate whether long-term, low-level exposure to monocyclic aromatic hydrocarbons (MAHs) induced insulin resistance. METHODS: The subjects were 110 male workers who were occupationally exposed to styrene, toluene, and xylene. One hundred and ten age-matched male workers who had never been occupationally exposed to organic solvents were selected as a control group. Cytokines, which have played a key role in the pathogenesis of insulin resistance, and oxidative stress indices were measured. Assessment of exposure to MAHs was performed by measuring their ambient levels and their urinary metabolites in exposed workers, and the resulting parameters between the exposed group and non-exposed control groups were compared. RESULTS: There was no significant difference in general characteristics and anthropometric parameters between the two groups; however, total cholesterol, fasting glucose, fasting insulin, and homeostasis model assessment of insulin resistance levels were significantly higher in the exposed group. Phenylglyoxylic acid levels showed significant association with tumor necrosis factor-alpha, total oxidative status, and oxidative stress index via multiple linear regression analysis. Further, there was a negative correlation between methylhippuric acid levels and total anti-oxidative capacity, and there was a significant relationship between MAHs exposure and fasting glucose levels, as found by multiple logistic regression analysis (odds ratio = 3.95, 95% confidence interval = 1.074-14.530). CONCLUSION: This study indicated that MAHs increase fasting glucose level and insulin resistance. Furthermore, these results suggested that absorbing the organic solvent itself and active metabolic intermediates can increase oxidative stress and cytokine levels, resulting in the changes in glucose metabolism and the induction of insulin resistance.
Cholesterol ; Cytokines ; Fasting ; Glucose ; Glyoxylates ; Homeostasis ; Humans ; Hydrocarbons, Aromatic ; Insulin ; Insulin Resistance ; Linear Models ; Logistic Models ; Male ; Mandelic Acids ; Occupations ; Oxidative Stress ; Solvents ; Styrene ; Toluene ; Tumor Necrosis Factor-alpha ; Xylenes

OBJECTIVES: In this study, we summarized the External Quality Assessment Scheme (EQAS) for the biological monitoring of occupational exposure to toxic chemicals which started in 1995 and continued until a 31st round robin in the spring of 2010. The program was performed twice per year until 2009, and this was changed to once a year since 2010. The objective of the program is to ensure the reliability of the data related to biological monitoring from analytical laboratories. METHODS: One hundred and eighteen laboratories participated in the 31st round robin. The program offers 5 items for inorganic analysis: lead in blood, cadmium in blood, manganese in blood, cadmium in urine, and mercury in urine. It also offers 10 items for organic analysis, including hippuric acid, methylhippuric acid, mandelic acid, phenylglyoxylic acid, N-methylformamide, N-methylacetamide, trichloroacetic acid, total trichloro-compounds, trans,trans-muconic acid, and 2,5-hexanedione in urine. Target values were determined by statistical analysis using consensus values. All the data, such as chromatograms and calibration curves, were reviewed by the committee. RESULTS: The proficiency rate was below 70% prior to the first round robin and improved to over 90% for common items, such as PbB and HA, while those for other items still remained in the range of 60-90% and need to be improved up to 90%. CONCLUSION: The EQAS has taken a primary role in improving the reliability of analytical data. A total quality assurance scheme is suggested, including the validation of technical documentation for the whole analytical procedure.
Acetamides ; Cadmium ; Calibration ; Consensus ; Dietary Sucrose ; Environmental Monitoring ; Formamides ; Glyoxylates ; Hexanones ; Hippurates ; Mandelic Acids ; Manganese ; Occupational Exposure ; Songbirds ; Sorbic Acid ; Trichloroacetic Acid