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.

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.

To analyze one case of multi-organ involvement of immunoglobulin G4-related disease and to review the related literatures.Through investigating the clinical data of one case of multi-organ involvement of immunoglobulin G4 (IgG4)-related disease,many related references were retrospected.IgG4-related disease and multi-organ involvement are used as the key words to search on Pubmed and Wanfang Database.According to the clinical manifestations,including serology,imaging,histopathology,responses to steroid therapy,a 65-year-old female patient was diagnosed as IgG4-related disease.The patient exhibited salivary gland swelling,lacrimal glands,and orbital soft tissues.And multiple organs such as pancreas,liver,bile duct,retroperitoneum,the lymph node,lung,paranasal sinuses,kidney and ureter were affected in the patient.Glucocorticoids are the mainstay of therapy and the recommended dose is 0.5-1.0 mg · kg-1 · d-1.Immunosuppressive agents need to be used in relapsing patients or in the patients with multi-organ involvement and serious complications.IgG4-related disease usually affects many systems and involves multiple organs,which is always misdiagnosed to other diseases.Both the correct diagnosis with multidisciplinary fields and the early treatment with glucocorticoids are very important.

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.

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

Humans ; Molecular Biology ; Nephrotic Syndrome ; genetics

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.

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.