Object To determine the content of glycyrrihizic acid obtained when each individual ingredient in MAHUANG DECOCTION was decocted separately and then mixing the extracts with boiling water in comparison with that obtained by decocting the total composition together as a whole in the traditional way. Methods The contents of glycyrrhizic acid was determined by HPLC. Hypersil C 18 column was used, with acetonitrile∶0.1% acetic acid (33∶67) as the mobile phase and detected at the wavelength of 254 nm. Results The average recovery of glycyrrhizic acid when separately decocted was 102.43%, RSD=2.65%, while that of decoction in whole was 99.41%, RSD=3.11%. Conclusion The method was simple and accurate and was not interfered by other constituents in the prescription.

Objective·To observe mitochondria permeability transition pore (mPTP) opening and apoptosis of H9c2 myocardial cell stimulated by lipopolysaccharide (LPS),and to explore the anti-apoptotic effect of combined application of cyclosporine A (CsA) and ryanodine (Rya).Methods·The H9c2 cells were divided into Control group,LPS group,LPS+CsA group,LPS+Rya group,and LPS+CsA+Rya group.The mPTP opening state,Ca2+ concentration within cell and mitochondrial,mitochondrial membrane potential (AΦm),cell apoptosis,expression of Bax and Bcl-2 at mRNA and protein levels,and activity of caspase 3 were determined respectively.Results·mPTP opened after being stimulated by LPS for 24 h,which increased the fluorescence intensity for Ca2+in cytosolic and mitochondria by 298％ and 231％ respectively,induced about 1/3 cell apoptosis,improved the activity of caspase 3 approximately twice,and enhanced expression ofBax mRNA (P=0.008).The combined use of CsA and Rya effectively inhibited mPTP opening,increased the enhancement of fluorescence intensity for Ca2+in both cytosolic and mitochondria,maintained normal AΦrn,reduced LPS-induced apoptosis,inhibited the activity of caspase 3,and decreased Bax mRNA expression level induced by LPS in the myocardial cells.Conclusion·mPTP plays an important role in in LPS-induced myocardial apoptosis,whereas the combination of CsA and Rya can alleviate it effectively.

AIM: To determine the contents of ephedrine、laetrile、glycyrrhizic acid and liquiritin in Huagai Powder traditional decoction and granule decoction(Herba Ephedrae,Semen Armeniacae Amarum,Radix et Rhizoma Glycyrrhizae,etc.) by HPLC. METHODS: An Agilent SB-C_(18) column was used for ephedrine,the mobile phase was acetonitrile-0.1% phosphoric acid(4∶96),detection wavelength at 207 nm;An Agilent XDB-C_(18) column was used for laetrile,the mobile phase was methanol-water(23∶77),detection wavelength at 215 nm;An Agilent XDB-C_(18) column was used for glycyrrhizic acid,the mobile phase was methanol-0.2 mol/L ammonium-acid(67∶33∶1),detection wavelength at 250 nm; An Agilent XDB-C_(18) column was used for liquiritin,the mobile phase was acetonitrile-0.5% acetic acid(20∶80),detection wavelength at 276 nm. RESULTS: There were discrepancies between traditional decoction and granules in the contents of four ingredients,the average contents in granules were more than those in the traditional decoction. CONCLUSION: The peaks of the four ingredients are(segregated)(effectively),and the method is simple,conveient,exact and has good reproducibility,the other ingredients do not have interferences for the determination.

Objective To conduct breath test with a relatively large number of subjects for new data regarding breath acetone in diabetes using a high accuracy and high data throughput breath acetone analyzer based on the cavity ringdown spectroscopy (CRDS) technique.Methods The CRDS breath analyzer was validated by standard acetone gas samples with various concentrations and golden standard gas chromatography-mass spectrometry (GC-MS).A total of 917 breath samples from 260 type 2 diabetic (T2D) patients and 30 healthy individuals were collected under each of 4 different conditions: fasting, 2 h post-breakfast, 2 h post-lunch, and 2 h post-dinner, and the samples were tested by the breath analyzer.Results The linear fitting curve of standard acetone samples with various concentration had good linearity (R=1, P＜0.05).The linear fitting of the results of GC-MS and CRDS was 0.98, suggesting that the obtained acetone concentrations using both methods were consistent.For the 260 T2D subjects, the exhaled breath acetone concentrations ranged from 0.0 to 10.6×10-6, while for the 30 healthy subjects, the breath acetone concentration ranged from 0.1 × 10-6 to 2.0× 10-6.The mean breath acetone concentration of the 260 T2D subjects was (1.5±1.l)× 10-6, which was 1.4 times of(1.1±0.5)×10-6 for the 30 healthy subjects.The mean breath acetone concentrations under the 4 conditions for the 260 T2D subjects ((1.6±1.2)×10-6, (1.4±1.0)×10-6~, (1.4±0.9)×10-6, and (1.4±1.2)× 10-6) were higher than that of the 30 healthy subjects ((1.3±0.3)×10-6, (1.0±0.6)×10-6, (1.0±0.6)×10-6, and (1.1±0.4)×10-6), respectively.No correlation was found between the breath acetone concentration and the blood glucose level of the T2D subjects and the healthy individuals.Conclusions The GC-MS validation confirms that the CRDS breath acetone analyzer is a reliable instrument for fast response and on-line breath acetone measurement.An elevated mean breath acetone concentration exists in T2D subjects.The relationship between breath acetone level and physiological parameters needs to be further investigated.

OBJECTIVE: To assess the value of copy number variation analysis based on next generation sequencing (CNV-seq) in prenatal diagnosis for women at advanced maternal age. METHODS: A prospective analysis was carried out for women who underwent amniocentesis at 18~36 weeks of gestation for fetal CNV-seq for advanced maternal age. RESULTS: For 1461 unrelated Chinese women with a singleton pregnancy, CNV-seq was performed for all samples successfully. The proportion of chromosomal abnormalities was 2.3% (34/1461), of which 44.12% were submicroscopic copy number variations (<5 Mb). CONCLUSION Pregnant women at an advanced maternal age should be informed for not only common trisomies but all pathogenic chromosomal aberrations. NGS was a sensitive and accurate approach for detecting CNVs.
Chromosome Aberrations ; Chromosome Disorders ; DNA Copy Number Variations ; Female ; Humans ; Maternal Age ; Pregnancy ; Prenatal Diagnosis ; Prospective Studies