2. Simultaneous determination of four non-cholesterol sterols in like desmosterol human plasma using gas chromatography-mass spectrometry with selective ion monitoring
Chinese Pharmaceutical Journal 2012;47(7):560-564
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.
3.Correlation between mutant mRNA expression of COL4A5 and phenotypes of XLAS females
Yunfeng WANG ; Jie DING ; Fang WANG
Chinese Journal of Nephrology 1997;0(01):-
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.
6.Study on the Symptomatic Characteristics of Chinese Medicine of 52 Patients with Pulmonary Hypertension
Yutao FANG ; Jie WANG ; Zhicheng JING
Chinese Journal of Information on Traditional Chinese Medicine 2006;0(10):-
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.
7.Effect of Serums Containing Xuefuzhuyu Capsules and Xuesetong Capsules on the Proliferation of Vascular Smooth Muscle Cells Induced by Lysophosphatidie Acid
Jie WANG ; Yingkun FU ; Suping FANG
Chinese Journal of Information on Traditional Chinese Medicine 2006;0(03):-
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.
8.Prenatal gene diagnosis of Alport syndrome.
Hong-wen ZHANG ; Jie DING ; Fang WANG
Chinese Journal of Pediatrics 2007;45(9):681-683
9.Effects of miR-409-5p on the proliferation and migration of hepatoma HepG2 cell line
Rui WANG ; Chenhuan YU ; Jie FANG
Practical Oncology Journal 2017;31(4):305-309
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.
10.Analysis of phenotypic heterogeneity of male patients with X-linked Alport syndrome from one family
Yanqin ZHANG ; Jie DING ; Fang WANG
Chinese Journal of Applied Clinical Pediatrics 2016;31(17):1318-1320
Objective In this study,the phenotype heterogeneity of 2 male patients with X-linked Alport syndrome from one family was analyzed and the likely reasons were discussed by reviewing the literature.Methods The clinical data at the time of diagnosis and during 5 years follow-up of 2-male patients with X-linked Alport syndrome from one family were collected.The α5 (Ⅳ) chain expression in the epidermal basement membrane was detected by indirect immunofluorescence method.COL4A5 gene mutations in skin fibroblasts and genomic DNA were detected by using reverse transcription polymerase chain reaction and direct sequencing and PCR sequencing methods from skin fibroblasts and genomic DNA,respectively.Results The diagnostic age of patient Ⅲ 1 was 14 years old.He had only microscopic hematuria,and proteinuria was negative.A negative α5 (Ⅳ) chain staining pattern was detected in his epidermal basement membrane.After 5 years follow-up without drug treatment,he was 19 years old,had persistent microscopic hematuria and normal renal function.The urinary microalbumin was 19.2-31.8 mg/L.The diagnosis age of patient Ⅱ 4 was 29 years old.The hematuria and proteinuria were found at 22 years old.He was treated with tripterygium wilfordii for 1 year.His disease progressed to an end stage of renal disease and he received hemodialysis therapy at 24 years old.He had the renal transplantation surgery at 29 years old,just 2 months before he came to hospital.And his renal function was restored.After 5 years follow-up,his urine examination and renal function were normal.Both patients had a missense mutation c.3650G > A(p.G1217D) in exon 41 in COL4A5 gene.Conclusions The different phenotypes of 2 male patients from one family with X-linked Alport syndrome were reported.The most possible reason for this is somatic mosaic variants in COL4A5 gene based on literature review.Physicians should be alert to phenotype heterogeneity in male X-linked Alport syndrome despite having the same gene mutation.