1.The comparative analysis and bias evaluation on the results of serum CK,AST and LDH between two different kinds of biochemical analysis system
International Journal of Laboratory Medicine 2015;(14):2031-2033
Objective To discuss the comparability and bias evaluation on the results of serum creatine kinase (CK ) ,aspartate aminotransferase (AST) ,lactate dehydrogenase (LDH) inspection results between the two different kinds of biochemical analysis system .Methods The control quality materials with two concentration were detected respectively for degree of precision ,and the assessment standard was from manufacturer introductions and laboratory setting .Then in accordance with American Clinical and Laboratory Standard Institute(CLSI) document EP9‐A2 ,100 patients serum CK、AST、LDH were detected .Afterwards the relative bias(SE% ) was calculated ,and the comparative analysis bias evaluation was judged according to the half of CLIA′88 allowed total error .Results The variable coefficients mainly conformed to the requirements ,and the experimental results were reliable .The oth‐ers mainly lower than the acceptable limit besides SE% for LDH higher than it .Conclusion It is necessary to execute comparative analysis and bias evaluation to ensure the accuracy and comparability if the same item is detected in two analysis system .
2.Effects of Wnt5A gene overexpression on cytoskeletal proteins of melanocytes
Qianya SU ; Tong LIN ; Qiyan ZHOU ; Lin PENG
Chinese Journal of Dermatology 2017;50(8):584-588
Objective To evaluate the effect of Wnt5A gene overexpression on cytoskeletal proteins of melanocytes after the plasmid containing the Wnt5A gene is transfected into primary melanocytes.Methods In vitro cultured primary human melanocytes were divided into three groups:blank control group receiving no treatment,negative control group transfected with endotoxin-free pcDNA3.1 (+)empty vector by Lipo3000 in Opti-MEM medium,Wnt5A plasnid group transfected with endotoxin-free pcDNA3.1 (+) vector containing the Wnt5A gene by Lipo3000 in Opti-MEM medium.After the transfection,quantitative PCR (qPCR) was performed to measure the mRNA expression of Wnt5A,ras-related C3 botulinum toxin substrate 1 (Rac1),filamentous actin (F-actin) and β-tubulin,Western blot analysis to determine the protein expression of Wnt5A,receptor tyrosine kinase like orphan receptor 2 (ROR2),Rac1,F-actin and β-tubulin,and an immunofluorescence assay (IFA) to observe the expression of cytoskeletal proteins.Results qPCR showed significant differences in the mRNA expression of the Wnt5A gene and its downstream genes Rac1 and F-actin among the Wnt5A plasmid group,negative control group and blank control group (F =1374.179,112.576,66.458,respectively,all P < 0.01),but there was no significant difference in the mRNA expression of β-tubulin among the three groups (P > 0.05).Additionally,the Wnt5A plasmid group showed significantly higher mRNA expression of Wnt5A,Rac1 and F-actin compared with the blank control group and negative control group (all P < 0.05).As Western blot analysis revealed,compared with the blank control group and negative control group,the Wnt5A plasmid group showed significantly higher Wnt5A protein expression (both P < 0.05),but significantly lower protein expression of Rac 1,ROR2 and F-actin (all P < 0.05).However,no significant difference in β-tubulin protein expression was observed among the three groups (P > 0.05).IFA showed no obvious difference in the fluorescence intensity of β-tubulin or F-actin between the Wnt5A group and the two control groups,but melanocytes showed larger size and increased number of dendrites,and the cytoskeleton changed dramatically with varying fluorescence intensity of F-actin,fuzzy texture,fractured or locally clustered tonofilaments in the Wnt5A group.Conclusion The overexpression of the Wnt5A gene in melanocytes can regulate the mRNA and protein expression of cytoskeletal proteins,nake melanocytes larger and more dendritic,and cause changes in the cytoskeleton,which may facilitate the transportation of melanosomes,and participate in the occurrence of hyperpigmented diseases.