ABSTRACT
<p><b>OBJECTIVE</b>To investigate low density lipoprotein receptor (LDLR) function and gene mutation in Chinese patients with familial hypercholesterolemia(FH).</p><p><b>METHODS</b>Lymphocytes were isolated from 10 ml anticoagulated peripheral blood of the patients, then a flow-cytometric method (FCM) with 1,1'-dioctadecyl-3,3,3', 3-tetramethylindocarbocyanine perchlorate labelled low density lipoproetin (DiI-LDL) was used to identify the function of LDLR on the surface of lymphocytes. Genomic DNA was isolated from whole blood of FH patients and analyzed by PCR-single strand conformation polymorphism (SSCP) and nucleotide sequencing methods.</p><p><b>RESULTS</b>Defects of binding and uptaking of LDLR were identified by FCM in 2 FH patients in one family, and their parents were examined in the present study. Then they were analyzed genetically. The detected mutation was a deletion of A, which caused a frame shift in codon 297 of exon 6 and introduced a beforehand stop codon in codon 369.</p><p><b>CONCLUSION</b>A novel mutation of LDL receptor gene was detected by the combination of FCM and PCR-SSCP methods.</p>
Subject(s)
Adult , Child , Child, Preschool , Female , Humans , Male , Base Sequence , China , Cholesterol , Blood , Cholesterol, HDL , Blood , DNA , Chemistry , Genetics , DNA Mutational Analysis , Family Health , Flow Cytometry , Genotype , Hyperlipoproteinemia Type II , Blood , Genetics , Lipoproteins, HDL , Blood , Molecular Sequence Data , Mutation , Pedigree , Phenotype , Polymorphism, Single-Stranded Conformational , Receptors, LDL , Genetics , Metabolism , Triglycerides , BloodABSTRACT
Objective To explore a practical way for traceability and comparability of the value obtained from clinical biochemical examination in regional medical organizations by transferring the assigned value in fresh serum to daily serum calibrator.Methods Six local comprehensive hospitals with Grade III were chosen to analyze the effects of calibration by traceability and the comparability of examined results.The definite values of standard panel in matching analytical system were transferred to fresh sera for value assignment.Verified biochemical analyzers were used to examine the values of alaninetransaminase(ALT),total cholesterol(TC),urea and total bilirubin(TBil) in mixed fresh sera and intercomparison with target system was performed to analyze the effect of traceable calibration and the comparability among various analytical systems.Results Although all the applied instruments in the present study were in good conditions with high precision,the comparability of obtained results of ALT,TC,Urea and TBil between tested system and target system was low and the differences were significant(P0.05).Conclusion Traceable precision transfer by using fresh serum is a simple,practical,feasible way to realize the traceability and comparability of regional clinical biochemical examination.