OBJECTIVE: To carry out genetic testing and prenatal diagnosis for a Chinese pedigree with Lesch-Nyhan syndrome (LNS) but no specimen from the affected probands. METHODS: All affected individuals in this pedigrees were male and had deceased during childhood, with no biological specimen left. Based on their typical neurological dysfunction and tendency for self-mutilation, the diagnosis of LNS was suspected. Sanger sequencing was carried out to detect potential variant of the HPRT1 gene among female members from the pedigree. Following the identification of the pathogenic variant, prenatal diagnosis was provided for a high-risk fetus. RESULTS: The proband's mother and three other females were found to harbor heterozygous c.500_501delGGinsC (p.Arg167fs*23) variant of the HPRT1 gene, which was unreported previously. Prenatal diagnosis showed that the fetus was a male and had inherited the same pathogenic variant. CONCLUSION The c.500_501delGGinsC variant of the HPRT1 gene probably underlay the LNS in this pedigree. Above finding has provided a basis for prenatal diagnosis and genetic counseling for this pedigree.
Male ; Female ; Humans ; Pregnancy ; Lesch-Nyhan Syndrome/genetics* ; Pedigree ; Hypoxanthine Phosphoribosyltransferase/genetics* ; Prenatal Diagnosis ; China ; Mutation

The prevalence of gout is increasing worldwide, and control of serum uric acid level has been regarded as one of the therapeutic methods for gout. Inhibition of xanthine oxidase (XO) activity which can oxidize hypoxanthine to uric acid has been commonly proposed to decrease serum uric acid level. The aim of this study was to demonstrate the hypouricemic effect of ethanol extract of Aster glehni leaves (EAG) by in vitro and in vivo study in potassium oxonate (PO)-induced hyperuricemic rats. EAG possessed 132.5 ± 6.8 mg QE/g of total flavonoid and showed antioxidant activity. EAG showed in vitro and in vivo inhibitory activity against XO and significantly decreased serum uric acid level in PO-induced hyperuricemic rats without liver toxicity. These results show that EAG significantly attenuates hyperuricemia by inhibiting XO activity, which resulted in the decrease of serum uric acid level. Therefore, EAG might possess a potential therapeutic ability for improving gout.
Animals ; Ethanol* ; Gout ; Hyperuricemia ; Hypoxanthine ; In Vitro Techniques ; Liver ; Potassium* ; Prevalence ; Rats* ; Uric Acid ; Xanthine Oxidase

PURPOSE: We sought to develop a matrix assisted laser desorption ionization-time of flight (MALDI-TOF)-based, ovarian cancer (OVC), low-mass-ion discriminant equation (LOME) and to evaluate a possible supportive role for triple-TOF mass analysis in identifying metabolic biomarkers. MATERIALS AND METHODS: A total of 114 serum samples from patients with OVC and benign ovarian tumors were subjected to MALDI-TOF analysis and a total of 137 serum samples from healthy female individuals and patients with OVC, colorectal cancer, hepatobiliary cancer, and pancreatic cancer were subjected to triple-TOF analysis. An OVC LOME was constructed by reference to the peak intensity ratios of discriminatory low-mass ion (LMI) pairs. Triple-TOF analysiswas used to select and identify metabolic biomarkers for OVC screening. RESULTS: Three OVC LOMEs were finally constructed using discriminatory LMI pairs (137.1690 and 84.4119 m/z; 496.5022 and 709.7642 m/z; and 524.5614 and 709.7642 m/z); all afforded accuracies of > 90%. The LMIs at 496.5022 m/z and 524.5614 m/z were those of lysophosphatidylcholine (LPC) 16:0 and LPC 18:0. Triple-TOF analysis selected seven discriminative LMIs; each LMI had a specificity > 90%. Of the seven LMIs, fourwith a 137.0455 m/z ion atretention times of 2.04-3.14 minuteswere upregulated in sera from OVC patients; the ion was identified as that derived from hypoxanthine. CONCLUSION: MALDI-TOF–based OVC LOMEs combined with triple-TOF–based OVC metabolic biomarkers allow reliable OVC screening; the techniques are mutually complementary both quantitatively and qualitatively.
Biomarkers ; Colorectal Neoplasms ; Female ; Humans ; Hypoxanthine ; Lysophosphatidylcholines ; Mass Screening ; Mass Spectrometry* ; Ovarian Neoplasms* ; Pancreatic Neoplasms ; Sensitivity and Specificity

BACKGROUND: Quantitative real-time polymerase chain reaction (qPCR) is the most reliable tool for gene expression studies. Selection of housekeeping genes (HKGs) that are having most stable expression is critical to carry out accurate gene expression profiling. There is no 'universal' HKG having stable expression in all kinds of tissues under all experimental conditions. METHODS: The present study aims to identify most appropriate HKGs for gene expression analysis in glioblastoma (GBM) samples. Based on literature survey, six most commonly used HKGs that are invariant in GBM were chosen. We performed qPCR using RNA from formalin fixed paraffin embedded GBM samples and normal brain samples to investigate the expression pattern of HPRT, GAPDH, TBP, B2M, B2M, RPL13A, and RN18S1 with different abundance. A simple Deltacycle threshold approach was employed to calculate the fold change. RESULTS: Our study shows that the expression of RPL13A and TBP were found to be most stable across all the samples and are thus suitable for gene expression analysis in human GBM. Except for TBP, none of the other conventionally used HKGs in GBM studies e.g., HPRT and GAPDH were found to be suitable as they showed variation in RNA expression. CONCLUSION: Validation of HKGs is therefore immensely specific for a particular experimental setup and is crucial in assessing any new setup.
Brain ; Formaldehyde ; Gene Expression Profiling ; Gene Expression* ; Genes, Essential* ; Glioblastoma* ; Humans ; Hypoxanthine Phosphoribosyltransferase ; Paraffin ; Real-Time Polymerase Chain Reaction* ; RNA

PURPOSE: Patients show variable responses to chemoradiotherapy (CRT), which is generally administered before surgery for locally advanced rectal cancer (LARC). The aim of this study was to identify molecular markers predictive of CRT responses by analysis of low-mass ions (LMIs) in serum of LARC patients. MATERIALS AND METHODS: LMIs (< 1,000 m/z) in serum obtained before CRT from 73 LARC (cT3-4) patients were profiled using matrix-assisted laser desorption/ionization mass spectrometry. LMIs with higher weighting factors in discriminating CRT responses were selected using principal components analysis and discriminant analysis. Selected LMIs were identified using the Human Metabolome Database. The concentrations of identified LMIs were determined by colorimetric enzyme assay, and compared according to post-CRT pathological stage (ypStage) or Dworak's tumor regression grade (TRG). RESULTS: The nine highest-ranking LMIs were selected. Among them, two LMIs with 137.08 and 169.04 m/z were identified as hypoxanthine (HX) and phosphoenolpyruvic acid (PEP), respectively. Higher HX concentration was observed in patients with ypStage 0-1 compared to ypStage 2-4 (p=0.034) or ypStage 3-4 (p=0.030); a similar difference was observed between TRG 4-3 and TRG 1 (p=0.035). HX > 16.0 muM showed significant association with ypStage 0-1 or TRG 4-3 than ypStage 3-4 (p=0.009) or TRG 1 (p=0.024), respectively. In contrast, a significantly lower concentration of PEP was observed in TRG 4-3 compared with TRG 2-1 (p=0.012). CONCLUSION: Findings of this study demonstrated that serum concentrations of HX and PEP, identified using LMI profiling, may be useful for predicting the CRT response of LARC patients before treatment.
Biological Markers* ; Chemoradiotherapy* ; Enzyme Assays ; Humans ; Hypoxanthine* ; Ions ; Mass Spectrometry ; Metabolome ; Phosphoenolpyruvate ; Rectal Neoplasms*

This study is to analyze and identify the water soluble components of water buffalo horn (Bubali Cornu, WBH), and also establish a method for investigating these components. Shotgun proteomic analysis identified proteins in WBH aqueous extraction: keratin, collagen, desmoglein, etc. Ultrafiltration and LC-MS/MS were used to separate and identify the peptides in WBH aqueous extract, as a result, identified peptides were mainly derived from nonspecific degradation products of keratin and collagen, which including C-terminal peptides and non C-terminal peptides. Hypoxanthine, uridine, guanosine, and adenosine were identified by comparing with the standards. The strategy in present study could be used in analyzing water soluble components of animal horn derived TCM. It provides a reference for investigation of the material basis of animal horn derived TCM.
Animals ; Buffaloes ; Chromatography, Liquid ; Guanosine ; Horns ; chemistry ; Hypoxanthine ; Mass Spectrometry ; Medicine, Chinese Traditional ; Peptides ; Proteomics ; Tandem Mass Spectrometry ; Uridine

Deficiency of hypoxanthine-guanine phosphoribosyltransferase is a purine nucleotide disorder and is the most common genetic cause of uric acid overproduction. This disease has a wide range of spectrum with regard to neurological features depending on the extent of the enzymatic deficiency. Complete deficiency of hypoxanthine-guanine phosphoribosyltransferase, called Lesch-Nyhan syndrome, is presented with hyperuricemia and characteristic neurological manifestation and self-mutilation. Partial hypoxanthine-guanine phosphoribosyltransferase--deficient patients are presented with a various intensities of the aforementioned symptoms, from almost normal neurologic manifestation to a severe form along with hyperuricemia. We report a twenty-year-old man with complete hypoxanthine-guanine phosphoribosyltransferase mutation and Lesch-Nyhan sydrome, who manifested gouty arthritis without neurologic symptom.
Arthritis, Gouty* ; Humans ; Hyperuricemia ; Hypoxanthine Phosphoribosyltransferase ; Lesch-Nyhan Syndrome* ; Neurologic Manifestations* ; Uric Acid

OBJECTIVETo study the chemical constituents of the leaves of Aquilaria sinensis.

METHODThe compounds were isolated and purified by the methods of solvent extraction and chromatographic technique, and their structures were identified on the basis of the analyses of spectral data.

RESULTThirty-three compounds were obtained. Among them, twelve compounds were identified as 5-hydroxyl-7,4'-dimethoxyflavone (1), acacetin (2), luteolin (3), genkwanin (4), yuankanin (genkwanin-5-O-beta-D-primeveroside, 5), adenosine (6), genkwanin-5-O-beta-D-glucopyranoside (7), hypolaetin-7-O-beta-D-glucopyranoside (8), hypoxanthine (9), uracil (10), 8-C-beta-D-galactopyranosylisovitexin (11), and 4-(1,2,3-trihydroxypropyl) -2,6-dimethoxyphenyl-1-O-beta-D-glucopyranoside (12), respectively.

CONCLUSIONAll compounds except for 1, 3 and 4 were isolated from the leaves of A. sinensis for the first time.

Adenosine ; analysis ; isolation & purification ; Chromatography ; methods ; Flavones ; analysis ; isolation & purification ; Glycosides ; analysis ; isolation & purification ; Hypoxanthine ; analysis ; isolation & purification ; Luteolin ; analysis ; isolation & purification ; Plant Extracts ; analysis ; isolation & purification ; Plant Leaves ; chemistry ; Thymelaeaceae ; chemistry

This study is to establish a method for simultaneously determination of five nucleosides and nucleobases, including hypoxanthine, uridine, adenine, guanosine and adenosine in Rehmannia glutinosa Libosch. which was collected from different regions in China. A Diamonsil C18 column (250 mm x 4.6 mm, 5 microm) was used. Acetonitrile and 0.04 mol L(-1) potassium dihydrogen phosphate solution were adopted as mobile phase with gradient elution. The flow rate was 1 mL min(-1) and column temperature was 30 degrees C. The detection wavelength was at 254 nm. The method had good linearity over the range of 1.0 - 16.0 microg mL(-1) (r2 = 0.999 8), 5.0 - 80.0 microg mL(-1) (r2 = 0.999 8), 1.0 - 16.0 microg mL(-1) (r2 = 0.999 5), 1.25 - 20.0 microg mL(-1) (r2 = 0.999 8) and 1.0 - 16.0 microg mL(-1) (r2 = 0.999 8) for hypoxanthine, uridine, adenine, guanosine and adenosine, respectively. The average recoveries were between 98.8% and 100.7%. The content of hypoxanthine, uridine, adenine, guanosine and adenosine in Rehmannia glutinosa Libosch. from different regions was significantly different. This established method was sensitive and reliable for the quantification of five chemical constituents in Rehmannia glutinosa Libosch.
Adenine ; analysis ; Adenosine ; analysis ; Chromatography, High Pressure Liquid ; Guanosine ; analysis ; Hypoxanthine ; analysis ; Nucleosides ; analysis ; Plants, Medicinal ; chemistry ; Rehmannia ; chemistry ; Uridine ; analysis

OBJECTIVE: To investigate the relationship between postmortem interval (PMI) and concentration changes of components in swine vitreous humor. METHODS: Ninety-six porcine eyes from swine dying from acute massive hemorrhage, being randomly divided into 24 groups, were stored in dark situation, at temperature of (15 +/- 2) degrees C and humidity of (50 +/- 5)% for 2-96 hours separately. The vitreous humor was collected. Concentrations of K+, Na+, Cl- and hypoxanthine (Hx) were analyzed by automatic biochemical analyzer and ultra performance liquid chromatograph (UPLC). The data were statistically analyzed by SPSS software. RESULTS: Linear regression analysis showed that concentrations of vitreous K+ and Hx were positively correlated with PMI(R2=0.767 and R2 = 0.793, respectively). Binary linear regression showed a higher correlation for K+ and Hx with PMI estimation (R2 = 0.866). PMI was not significantly correlated with vitreous Na+ and Cl- concentrations. CONCLUSION Vitreous K+ and Hx concentrations can be used as the objective markers for PMI estimation. The binary linear regression functions of vitreous K+ and Hx concentrations with PMI are more accurate for estimating the PMI.
Animals ; Chromatography, High Pressure Liquid/methods* ; Female ; Forensic Pathology ; Hypoxanthine/analysis* ; Male ; Postmortem Changes ; Potassium/analysis* ; Regression Analysis ; Sodium/analysis* ; Swine ; Temperature ; Time Factors ; Vitreous Body/chemistry*