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.
4.Correlation between primary hypertension and benign prostatic hyperplasia in the elderly
Jie CAO ; Haiya WANG ; Ningyuan FANG
Chinese Journal of Geriatrics 2013;(2):165-167
Objective To explore the relationship between primary hypertension and benign prostatic hyperplasia (BPH) in the elderly.Methods From August 2010 to June 2012,135 consecutively referred patients with BPH were enrolled in this study,in which 70 cases were BPH with hypertension,65 cases were isolated benign prostatic hyperplasia.All patients were questioned in detail about history,body weight and height were measured to calculate the body mass index (BMI).Prostate specific antigen (PSA) and fasting plasma glucose (FPG) and lipids were assayed in the study.The international prostate symptom score (IPSS) were recorded.Prostate volume (PV) was detected by abdominal ultrasound.Results The levels of IPSS and PV were higher in BPH with hypertension than simple BPH group [(18.9±7.5)scores vs.(13.2±7.8) scores,P<0.05 ;(43.0±15.4) ml vs.(39.2 ± 13.9) ml,P< 0.05].Pearson analysis showed that PV was positively correlated with age and the years of hypertension (r=0.34,0.29,P<0.05).After adjustment for age,PV was significantly greater in hypertension group with more than 15 years compared to less than 15 years of hypertension group.Conclusions Hypertension,particularly long-term hypertension is related to BPH in the elderly.The long term of hypertension may accelerate the occurrence and clinical progression of BPH.
5.Analysis of clinical characteristics of benign prostatic hyperplasia and the correlation between BPH and cardivascular diseases in the elderly
Jie CAO ; Haiya WANG ; Ningyuan FANG
Chinese Journal of Geriatrics 2010;29(11):894-895
Objective To realize the clinical characteristics and treatment strategies in elderly patients with benign prostatic hyperplasia (BPH), and investigate the correlation between severity of BPH and cardiovascular diseases. Methods One hundred consecutively referred patients with BPH were enrolled in this study, and the international prostate symptom score (IPSS) and quality of life (QOL) scores were recorded. All patients were queried in detail about history of cardiovascular disease, and underwent detection of prostate specific antigen (PSA) levels, prostate volume (PV)measurement by abdominal ultrasound. Results PV and serum PSA level increased with age.Forty-eight percent of patients had a moderately enlarged prostate (IPSS 8-19). Patients with BPH had higher incidence of hypertension, diabetes, as well as coronary artery disease (P<0.05). The most common medical treatments were 5α-reductase inhibitors and a-receptor blockers in our hospital and most patients had good compliance. Conclusions The severity of BPH is correlated with age and morbidity of coronary artery disease. For the drug intervention therapy, 5a-reductase inhibitors have the highest utilization rate.
6.Prenatal gene diagnosis of Alport syndrome.
Hong-wen ZHANG ; Jie DING ; Fang WANG
Chinese Journal of Pediatrics 2007;45(9):681-683
9.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.
10.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.