Adolescent ; Child ; Electroencephalography ; Female ; Fructose ; analogs & derivatives ; therapeutic use ; Humans ; Male ; Migraine Disorders ; diagnostic imaging ; drug therapy ; physiopathology ; Ultrasonography, Doppler, Transcranial

OBJECTIVE: To establish a method for simultaneous determination of phytosterols and cholesterol precursors in human plasma by GC-MS to understand the metabolism of cholesterol in patients. METHODS: Sterols as TMS derivatives were separated and identified on HP-5MS column (30 m × 0.25 μm × 0.25 mm). The initial column temperature of 200°C. was held for 2 min, then increased at a rate of 15°C · min-1 to 280°C and maintained at this temperature for further 30 min. The internal standard was 5α-cholestane. The method was based on the alkaline hydrolysis of sterol esters, extraction of free sterols and derivatization. RESULTS: The calibration curves of the sterols were linear. Inter-day and intra-day RSDs were lower than 7.9% and 9.8%. The recoveries based on replicate analyses of plasma samples spiked with known amounts of sterol standards were between 89.5% - 106.3%. Plasma samples of 20 patients with hyperlipemia were determined by the method. The mean values of non-cholesterol sterols were: desmosterol (0.39 ± 0.08) mg · L-1, lathosterol (3.41 ± 0.83) mg · L-1, campesterol (3.12 ± 0.53) mg · L-1, and sitosterol (4.22 ± 0.67) mg · L-1. CONCLUSION: An optimized analytical method for simultaneous analysis of cholesterol, desmosterol, lathosterol, campesterol and sitosterol in plasma has been developed using capillary gas chromatography coupled to mass spectrometry (GC-MS) with multiple selected ion monitoring (SIM). With good accuracy and precision, it is suitable for the analysis of the individual difference of cholesterol metabolism. Copyright 2012 by the Chinese Pharmaceutical Association.

Objective To understand the different phenotype based on COL4A5 mutations and to analyze the correlation between mutant mRNA expression of COL4A5 gene and phenotype variability of XLAS females. Methods Skin biopsy specimens were obtained from 6 XLAS females with confirmed COL4A5 deletion mutations. Total RNA was isolated from cultured skin fibroblasts and RT-PCR was performed to amplify the fragment including the mutation sequences of COL4A5 gene. The PCR products were electrophoresed with 8% polyacrylamide gel. Each fragment of PCR product revealed under polyacrylamide gel was further analyzed with the optical absorptance (A). The A ratio of mutant over total mRNA of COL4A5 gene (total mRNA means the mutant mRNA plus normal mRNA) was analyzed with different phenotypes. Results Besides microscopic hematuria, the females often had gross hematuria and persistent and severe proteinuria when the mutant a5 (IV) mRNA expression was more than the normal. Whereas the XLAS females didn't had gross hematuria and proteinuria or had discontinuous and mild proteinuria when the mutant ?5 (Ⅳ) mRNA expression was less than the normal. The proteinuria was positively correlated with the expression ratio of mutant over total mRNA of COL4A5 gene in the females. Conclusions The quantity of the mutant mRNA expression of COL4AS gene is correlated with the phenotypic severity of XLAS females. The XLAS females with much more mutant mRNA expression of COL4A5 gene show more severe phenotypes of AS.

Adolescent ; Heart Valve Diseases ; etiology ; Heart Valves ; pathology ; Humans ; Lupus Erythematosus, Systemic ; complications ; pathology ; Male

Objective To study the symptomatic characteristics distribution of Chinese medicine of patients with pulmonary hypertension.Methods Symptomatic information of 52 patients with pulmonary hypertension was collected to find the symptomatic characteristics of Chinese medicine,and analyse the correlation of symptom and pulmonary hypertension with different types and degrees.Results The proportion of TCM symptom of pulmonary hypertension from high to low were syndrome of strong Qi sinking(96.2%),syndrome of blood stasis(75.0%),syndrome of deficiency of lung Qi(42.3%),syndrome of deficiency of heart Qi(30.8%),syndrome of deficiency of spleen Qi(17.3%).Syndrome of blood stasis was the most closely related to the congenital cardiopathy(CC) with pulmonary hypertension.Syndrome of deficiency of heart Qi was the most closely related to the degree of pulmonary hypertension increased pressure.The next one was syndrome of deficiency of lung Qi.Conclusion Syndrome of strong Qi sinking,syndrome of blood stasis,syndrome of deficiency of heart Qi and syndrome of deficiency of lung Qi are the main syndromes of pulmonary hypertension.Syndrome of strong Qi sinking is the most closely related to the pulmonary hypertension.

Objective To investigate the effect of the serums containing Xuefuzhuyu capsules and Xuesetong capsules on the proliferation of vascular smooth muscle cells (VSMC) induced by 1ysophosphatidie acid (LPA). Methods Rat VSMCs were obtainded by tissue-piece inoculation and divided into seven groups:blank serum group, Xuefuzhuyu (Xuesetong) capsules-containing serum group, LPA+ Xuefuzhuyu (Xuesetong, captopril) containing serum group and LPA+blank serum group. Effects of different groups on the proliferation of VSMCs was evaluated by MTT colorimetry. Analysis of cell cycle and DNA content were performed by flow cytometry. Results The optical density (OD) and the S phase fraction (SPF) of LPA+blank serum group and Xuefuzhuyu capsules-containing serum group were more increased than the blank serum group (P 0.05, P 0.05). Conclusion Xuefuzhuyu capsules and LPA promoted the proliferation of VSMCs in vitro. Xuefuzhuyu and Xuesetong capsules can inhibit the proliferation induced by LPA, and effect of Xuesetong capsules was greater. Part of the mechanism may be associated with the inhibition of DNA synthesis and preventing the transition from G0/G1 to S phase of VSMCs.

Objective The objective of this study was to observe effects of miR-409-5p on the proliferation and migration of HepG2 cells.Methods HepG2 cells were transfected with miR-409-5p mimics and verified by Real-Time quantitative PCR.After high expression of miR-409-5p,MTT and wound healing assays were used to detect the proliferation and migration ability of HepG2 cells.Results The cell viability of HepG2 cells transfected with miR-409-5p mimics was significantly decreased in comparison with the control group and showed a dose-and time-dependent manner(P<0.05).The migration ability of cells overexpressing miR-409-5p mimics was significantly lower than that in the NC group(P<0.05).There was significant difference between the two groups at treatments for 24 h and 48 h.Conclusion Up-regulated miR-409-5p can inhibit the proliferation and migration of HepG2 cells.

Objective To explore the correlation between phenotypes in female with X-linked Alport syndrome(XLAS) and X-inactivation of different tissues.Methods Thirty-six female diagnosed as XLAS were studied,and proteinuria was taken as a marker of the severity of clinical phenotypes.X-inactivation patterns were analyzed in peripheral blood cells of 36 XLAS female and in skin fibroblasts of 12 XLAS female.The X-inactivation analysis was performed by using Hpa Ⅱ predigestion of DNA followed by polymerase chain reaction(PCR) of the highly polymorphic CAG repeat of the androgen receptor gene.Results The average X-inactivation levels of the mutant allele decreased while the degree of proteinuria increased,so there was a negative correlation between the degree of proteinuria and the X-inactivation ratios of the mutant allele in blood cells(r=-0.543,P=0.002).However,there was no correlation between the degree of proteinuria and the X-inactivation ratios of the mutant allele in skin fibroblasts(r=-0.131,P=0.701).Conclusions X-inactivation ratios might explain partially the diverse phenotypes in XLAS female patients,which suggested that the prognosis of XLAS female might be predicted via analysis of the X-inactivation in peripheral blood cells.

Humans ; Molecular Biology ; Nephrotic Syndrome ; genetics

Collagen Type IV ; genetics ; Female ; Fetal Development ; Humans ; Nephritis, Hereditary ; diagnosis ; genetics ; Pregnancy ; Prenatal Diagnosis ; methods