Construction of eukaryotic expression vector of LYRM1, a novel gene related with human obesity, and establishment of its stable transfected 3T3-L1 cell line / 浙江大学学报·医学版

<p><b>OBJECTIVE</b>To construct the eukaryotic expression vector of LYRM1 and to transfect it to preadipocytes cell line 3T3-L1.</p><p><b>METHODS</b>The complete coding sequence of LYRM1 gene was amplified by RT-PCR from human omental adipose tissue and was subcloned into eukaryotic expression vector pcDNA(TM)3.1/myc-His B. The recombinant plasmid was then transfected into 3T3-L1 preadipocytes by liposome. After screening culture by G418, stable transfected 3T3-L1 cell line was established. The expression of LYRM1 protein was identified by Western blot.</p><p><b>RESULT</b>The recombinant plasmid was confirmed by PCR, restriction enzyme digestion and sequencing. The stable transfected 3T3-L1 cell line was established and the LYRM1 protein was expressed successfully.</p><p><b>CONCLUSION</b>The eukaryotic expression vector of LYRM1 has been successfully established and the stably transfected 3T3-L1 cell line also established.</p>

Clinical characteristics and ultrastructural features of livers in children with Wilson disease manifested mainly as hepatic injuries / 中华肝脏病杂志

<p><b>OBJECTIVES</b>To study the feasibility and possibility to diagnose Wilson disease with electronmicroscopical examination of liver biopsies.</p><p><b>METHODS</b>Clinical analysis, histological observation and ultrastructural examination were performed on 15 children with Wilson disease.</p><p><b>RESULTS</b>All 15 subjects had symptoms of hepatic disorders, such as jaundice. Morphological signs of hepatocyte injury in three phase, namely steatosis, mitochondrion changes and cholestasis in bile canaliculi of the early phase, nucleus injury, dilation of endoplasmic reticulum, increase of lysosomes and appearance of residual bodies of the second phase, and massive autophagy and cirrhosis of the late phase were shown. A few inflammatory cells in the liver specimens were observed. Accumulation of copper in lysosomes and autophagosomes was found by energy-dispersion X-ray.</p><p><b>CONCLUSION</b>The diagnostic signs for Wilson disease are autophagosomes in hepatocytes, cirrhosis accompanied with a few of inflammatory cells. A certain diagnosis of the disease depends on the finding of copper accumulation in hepatocytes.</p>