ABSTRACT
A new polyhydroxycholestane sulfate ester, 3α,12ß,25,26-tetrahydroxy-7-oxo-5ß-cholestane 26-O-sulfate (1), was isolated from dried skin of Bufo bufo gargarizans Cantor and its structure was elucidated on the basis of extensive 1D and 2D NMR as well as HR-ESI-MS analysis. A comparison of steroidal metabolite profiles, based on HPLC and LC-MS analyses, indicates that the chemical compositions of the various parts of toads, such as venom, skin and stratum corneum, are significantly different.
Subject(s)
Bufanolides/isolation & purification , Bufo bufo , Cholestanols/isolation & purification , Animals , Bufanolides/chemistry , China , Cholestanols/chemistry , Chromatography, High Pressure Liquid , Molecular Structure , Nuclear Magnetic Resonance, Biomolecular , Skin/chemistry , VenomsABSTRACT
OBJECTIVE: To investigate the main factors that influence the results of sperm alkaline single-cell gel electrophoresis (SCGE), optimize the conditions, and standardize its procedures. METHODS: Using alkaline SCGE, we detected the DNA fragments of sperm treated with different concentrations of H2O2 and determined the influences of the number of agarose gel layers, pH during DNA unwinding and electrophoresis, the time of DNA unwinding and electrophoresis, and cumulative sperm number on the results of sperm alkaline SCGE. Then we optimized the procedures, analyzed the repeatability of the optimized method, and examined 40 semen samples using the method. RESULTS: Three agarose gel layers could reduce the background. The optimal pH during DNA unwinding and electrophoresis was 10, and the best times for DNA unwinding and electrophoresis were 40 min and 30 min, respectively. Fifty sperm were adequate to ensure the reliability of the results. Based on the percentage of tail DNA, the intra- and inter-assay repeatabilities of the optimized sperm alkaline SCGE were 3.12% and 7.13%, and by the DNA damage score, they were 2.38% and 6.09%, respectively. Sperm DNA fragments were significantly increased in the infertile patients with oligoasthenoteratozoospermia as compared with healthy fertile males (P <0.05). CONCLUSION: The optimized sperm alkaline SCGE, highly repeatable and easy to be standardized, can be applied to the clinical detection of sperm DNA fragmentation in infertile men.
Subject(s)
Asthenozoospermia/genetics , Comet Assay/standards , DNA Fragmentation , Oligospermia/genetics , Spermatozoa/enzymology , DNA Damage , Humans , Hydrogen Peroxide/toxicity , Male , Oxidants/toxicity , Reproducibility of Results , Sperm Count , Spermatozoa/drug effects , Time FactorsABSTRACT
OBJECTIVE: Diagnosing congenital hypothyroidism (CH) early was studied in this paper through neonatal screening to prevent children's mental retardation. METHODS: The first step is mensurating the concentration of thyroid stimulating hormone (TSH) with the method of time-distinguished fluorometry immunoassay. If the result is positive, then it is necessary to mensurate the serum TSH, free triiodothyronine (FT3),free thyroxin (FT4) with chemiluminescence immunoassay (CLIA) and execute clinical diagnosis. RESULTS: There were 68 confirmed congenital hypothyroidism patients among the 106,224 samples from Apr.1997 to Dec.2003 in Hunan province. The positive rate was 1/1,562, which was obviously higher than the national average value (1/3,009). CONCLUSION: First, neonatal screening is the key to prevent mental retardation through early diagnosing and treating CH. Second, the probability of suffering CH is high in Hunan province. Third, the positive rate is relative to the development of science and technology, while the accumulated rate is better to reflect the real incidence of the disease. Fourth, selecting proper critical value will help to prevent missing screening.