ABSTRACT
MicroRNAs (miRNAs) are a family of non-coding RNA that are able to adjust the expression of many proteins, including ATP-binding cassette transporter and organic cation transporter. We sought to evaluate the effect of miR-511 on the regulation of OATP1B1 expression by free fatty acids. When using free fatty acids to stimulate Chang liver cells, we found that the expression of miR-511 increased significantly while the expression of OATP1B1 decreased. We also proved that SLCO1B1 is the target gene of miR-511 with a bioinformatics analysis and using the dual luciferase reporter assay. Furthermore, the expressions of SLCO1B1 and OATP1B1 decreased if transfecting Chang liver cells with miR-511, but did not increase when transfecting the inhibitors of miR-511 into steatosis cells. Our study indicates that miR-511 may play an important role in the regulation of OATP1B1 expression by free fatty acids.
Subject(s)
Humans , Computational Biology , Fatty Acids, Nonesterified , Liver , Luciferases , MicroRNAs , RNA, UntranslatedABSTRACT
<p><b>OBJECTIVE</b>The objective of this research is to construct a clinic-usable genechip method for detection of hepatitis B virus lamivudine-resistant mutants and basal core promotor/Pre-C mutants, compare this method with DNA sequencing to investigate this genechip's character (sensity, specificity, stability and practicability in clinic) and apply it in clinic.</p><p><b>METHODS</b>This genechip detection method can detect the DNA and 8 mutative site of HBV, include 3 lamivudine-resistant mutation site(No. 180, 204, 207 site in DNA polymerase gene), 5 HBeAg escape-related mutation site (nt 1896, 1899, 1862, 1764,1762 site in BCP/Pre-C region).The results of genechip method was verified by DNA sequencing.</p><p><b>RESULTS</b>In detecting HBV DNA, the results of genechip were agree with 100% of the results of DNA sequencing. In detecting HBV mutants, 251 sites (in 32 samples, 256 sites) showed the same results using both methods, and only 5 sites were not completely match (P > 0.05). In these 5 sites, genechip methods got multi-infection results, but sequencing got single-infection results.</p><p><b>CONCLUSION</b>These results suggest that genechip method has the same positive rate and almost these same specificity with DNA sequencing method, and is better than DNA sequencing method in detecting multi-infected HBV strains. [Key words]</p>
Subject(s)
Humans , Antiviral Agents , Pharmacology , Base Sequence , Drug Resistance, Viral , Hepatitis B , Drug Therapy , Virology , Hepatitis B Core Antigens , Genetics , Hepatitis B virus , Genetics , Lamivudine , Pharmacology , Molecular Sequence Data , Mutation , Oligonucleotide Array Sequence Analysis , Methods , Promoter Regions, GeneticABSTRACT
<p><b>BACKGROUND</b>To establish a genechip method for detection of hepatitis B virus (HBV) DNA, basal core promotor (BCP), and Pre-C mutants.</p><p><b>METHODS</b>This study used two kinds of technology (PCR, oligochip), which can detect five mutant hotspots including nt 1 896, nt 1 899, nt 1 862, nt 1 764 and nt 1 762. The results of genechip method was verified by DNA sequencing.</p><p><b>RESULTS</b>In detecting HBV DNA, the results of genechip were 100% consistent with those of DNA sequencing. In detecting HBV BCP and Pre-C mutants, 146 codons showed the same results using both methods, except for only 4 codons (P greater than 0.05).</p><p><b>CONCLUSION</b>This convenient high throughput genechip method could detect several BCP and Pre-C mutant codons at the same time. These results suggest that genechip method has the same positive rate and specificity with DNA sequencing method. It has more advantages than the latter in detecting mixed mutants and therefore may be used in clinical practice.</p>
Subject(s)
Humans , DNA, Viral , Hepatitis B , Virology , Hepatitis B virus , Genetics , Mutation , Oligonucleotide Array Sequence Analysis , Methods , Polymerase Chain Reaction , Methods , Promoter Regions, Genetic , Sequence Analysis, DNA , Viral Core Proteins , GeneticsABSTRACT
<p><b>OBJECTIVE</b>To establish an advanced purification techniques of the essential oils obtained from Pogostemon cablin.</p><p><b>METHOD</b>Molecular distillation (MD) was applied.</p><p><b>RESULT</b>Four distillates were obtained, chemical constituents of which were analyzed with GC-MS. Compared with those in original oils, the contents of active compounds (patchouli alcohol and pogostone) rose by 27%-47% in the distillates II and III.</p><p><b>CONCLUSION</b>Molecular distillation (MD) effectively raises the contents of patchouli alcohol and pogostone. The work is of great economic and scientific significance for the industrialization of P. cablin and the discovery of new drugs.</p>
Subject(s)
Gas Chromatography-Mass Spectrometry , Lamiaceae , Chemistry , Oils, Volatile , Chemistry , Plants, Medicinal , Chemistry , Sesquiterpenes , Technology, Pharmaceutical , MethodsABSTRACT
<p><b>OBJECTIVE</b>Lamivudine resistant HBV strains in Shenzhen were detected at multiple sites and in large amounts to understand further the distribution of lamivudine resistant mutants.</p><p><b>METHODS</b>552 Hepatitis B patients's sera were examined using genechip method. Among them, 192 samples of lamivudine resistant mutant were further analyzed.</p><p><b>RESULTS</b>In those 192 lamivudine resistant samples, 191 were YMDD mutants, 124 mutants of codon 528 and 9 mutants of codon 555. 88% YMDD mutants were multi-mutants of YVDD and codon 528; single mutants of YIDD; multi-mutants of YIDD and codon 528. 91% codon of YMDD mutants were GTG, ATT; the other 9% were ATA, ATC.</p><p><b>CONCLUSIONS</b>These results suggest that mutants of codon 552 (YMDD) are core mutants. Mutants of codon 528 and 555 are incidental mutants, YVDD mutants always emerge with mutants of codon 528, but YIDD mutants appear differently. 9% YMDD mutants's codons are ATA or ATC. This may be the reason for the low positive rate shown by using the conventional PCR methods.</p>