ABSTRACT
A quantitative analytical method based on HPLC coupled with the charged aerosol detector (CAD) for quantitative analysis of multi-components with a single marker (QAMS) was established for simultaneous determinations of astragaloside Ⅰ, astragaloside Ⅱ, astragaloside Ⅳ, calycosin-7-O-β-D-glucoside, formononetin and 7,2'-dihydroxy-3',4'-dimethoxyisoflavan in Astragalus membranaceus. The separation was performed on an Agilent SB-C18 (150 mm×4.6 mm, 3.5 μm), with gradient elution using the mobile phase consisting of 0.05% formic acid solution and 0.05% formic acid acetonitrile at the flow rate of 1.0 mL·min-1. The column temperature was 35 ℃, and the injection volume was 20 μL. For CAD, the drift tube temperature was at 50 ℃. The contents of six components in A. membranaceus were determined by both external standard method (ESM) and QAMS, and then were compared. The results showed that chromatographic peaks were separated well and the linear ranges of astragaloside Ⅰ, astragaloside Ⅱ, astragaloside Ⅳ, calycosin-7-glucoside, formononetin and 7,2'-dihydroxy-3',4'-dimethoxyisoflavan were 0.113-2.250 mg·mL-1, 0.012-0.240 mg·mL-1, 0.004-0.080 mg·mL-1, 0.065-1.300 mg·mL-1, 0.005-0.100 mg·mL-1 and 0.007-0.150 mg·mL-1, respectively. The content ranges of astragaloside Ⅰ, astragaloside Ⅱ, astragaloside Ⅳ, calycosin-7-glucoside, formononetin and 7,2'-dihydroxy-3',4'-dimethoxyisoflavan were 0.306-0.922 mg·g-1, 0.053-0.183 mg·g-1, 0.015-0.092 mg·g-1, 0.069-0.823 mg·g-1, 0-0.098 mg·g-1 and 0.020-0.107 mg·g-1 in 20 batches of A. membranaceus, respectively. Using astragaloside Ⅱ as an internal reference, the relative correlation factors of astragaloside Ⅰ, astragaloside Ⅳ, calycosin-7-O-β-D-glucoside, formononetin, and 7,2'-dihydroxy-3',4'-dimethoxyisoflavan were calculated as 0.561, 0.835, 0.299, 0.796, and 0.799, respectively. The results were compared with those obtained by the external standard method to verify the feasibility, rationality and repeatability of QAMS method, and there was no significant difference in assay results between the two methods. In conclusion, the QAMS method is accurate and feasible, and could be used to determine the contents such as astragaloside Ⅰ, astragaloside Ⅱ, astragaloside Ⅳ, calycosin-7-glucoside, formononetin and 7,2'-dihydroxy-3',4'-dimethoxyisoflavan, and it can be used for quality control of A. membranaceus.
ABSTRACT
<p><b>OBJECTIVE</b>To identify potential mutation of human androgen receptor (AR) gene in a patient with complete androgen insensitivity syndrome (CAIS).</p><p><b>METHODS</b>DNA sequences of 8 exons and exon/intron boundaries of the AR gene were amplified with PCR and directly sequenced.</p><p><b>RESULTS</b>DNA sequencing has revealed a frameshift mutation due to deletion of nucleotide C at position 3507 in exon 6, which gave rise to a stop codon resulting premature termination for translation.</p><p><b>CONCLUSION</b>A novel frameshift mutation in exon 6 of AR gene probably underlies the disease in our patient.</p>
Subject(s)
Humans , Male , Young Adult , Androgen-Insensitivity Syndrome , Diagnosis , Genetics , Base Sequence , Exons , Frameshift Mutation , Phenotype , Receptors, Androgen , GeneticsABSTRACT
Objective: To investigate the inhibitory effect of short hairpin RNA (shRNA) -mediated silence of phospholipase C epsilon (PLC)̇gene expression on the proliferation of renal carcinoma 786-0 cells and its action mechanism. Methods: Liposome was employed to mediate the transfection of both the recombinant plasmid (pGenesil-PLC)̇and the control plasmid (pGenesil-NP) into the 786-0 cells. RT-PCR was used to detect the PLĊ mRNA expression after being transfected for 48 h. MIT assay was conducted to detect the proliferation inhibitory rate at 24, 48, and 72 h after transfection, respectively. Flow cytometry (FCM) was performed to analyze cell cycle after 48-h transfection. RT-PCR and immunocytochemistry were used to analyze the expression levels of both p27 and Ki67. Results: The expression of PLĊmRNA was significantly inhibited by recombinant plasmid transfection with the inhibitory rate of 69.7%. The proliferation inhibitory rates were 21.2%, 31.6%, and 32.7% after being transfected for 24, 48, and 72 h, respectively. FCM analysis demonstrated that the distribution of cell cycle changed. The number of cells in the G0/G1 phase increased, and that in both the S phase and G2/M phase decreased. Cells were arrested at the G0/G1 phase, and the subdiploid "apoptotic peak" appeared at the same time. RT-PCR and immucytochemistry indicated that the expression of p27 was up-regulated and that of Ki67 was downregulated. Conclusion: The proliferation of the renal carcinoma 786-0 cell line is inhibited by interference of PLĊgene expression, and the underlying mechanism may partially be related with up-regulation of P27 and down-regulation of Ki67.
ABSTRACT
Objective To investigate the relationship between high-sensitive C-reactive protein(hsCRP)and insulin resistance(IR)in elderly hypertension patients.Methods The levels of ambulatory blood pressure mo- nitoring(ABPM),fasting plasma glucose(FPG),fasting insulin(FINS),von Willebrand factor(vWF),hsCRP were measured in elderly patients with hypertension alone(n=260),hypertension coexit with diabetes(n=230), and healthy matched subjects(n=250).Results The levels of FINS,FPG,vWF,hsCRP,systolic blood pres- sure(SBP),diastolic blood pressure(DBP)in the HT and HT+DM group were significantly higher than that in the NC group(P