Wilson's disease, an autosomal recessive disorder, is characterized by the excessive accumulation of copper in the liver. WND (ATP7B) gene, which encodes a putative copper transporting P-type ATPase, is defective in the patients. To investigate the in vivo function of WND protein as well as its intracellular localization, WND cDNA was introduced to the Long-Evans Cinnamon rat, known as a rodent model for Wilson's disease, by recombinant adenovirus-mediated gene delivery. An immunofluorescent study and a subcellular fractionation study revealed the transgene expression in liver and its localization to the Golgi apparatus. Moreover, since the synthesis of holoceruloplasmin is disturbed in the LongEvans Cinnamon rat, the plasma level of holoceruloplasmin, oxidase-active and copper-bound form, was examined to evaluate the function of WND protein with respect to the copper transport. Consequently, the appearance of holoceruloplasmin in plasma was confirmed by Western blot analysis and plasma measurements for the oxidase activity and the copper content. These findings indicate that introduced WND protein may function in the copper transport coupled with the synthesis of ceruloplasmin and that the Golgi apparatus is the likely site for WND protein to manifest its function.Wilson's disease, an autosomal recessive disorder, is characterized by the excessive accumulation of copper in the liver (1). This phenomenon is thought to be due to reduced biliary excretion of copper and disturbed incorporation of copper into ceruloplasmin (CPN).1 Hepatic copper at toxic levels causes liver cirrhosis, and extrahepatic copper toxicity occurs especially in the brain due to the released copper from the damaged liver. WND (officially designated ATP7B), identified as the gene responsible for this disease, encodes a putative copper transporting P-type ATPase (2-5). The observations of single base changes or small deletions within WND of Wilson's disease patients have already been reported (3, 6).The Long-Evans Cinnamon (LEC) rat, known as an animal model for Wilson's disease, shows some of the clinical features similar to Wilson's disease, including hepatic copper accumulation, reduced biliary copper excretion, reduced copper in plasma, and a remarkable decrease of serum CPN activity (7,8). Atp7b, the rat gene homologous to WND, has been cloned, and a partial deletion at the 3Ј end in this gene is reported in the LEC rat (9). It is also known that the expression of Atp7b mRNA is absent in the LEC rat (10).CPN, a blue copper oxidase in plasma, contains 90 -95% of plasma copper. This protein is synthesized mainly in hepatocytes and secreted into plasma with 6 atoms of copper per molecule as the oxidase active holoprotein (11-13). The reduced levels of oxidase activity of CPN in the circulation of Wilson's disease patients and LEC rats is due to the secretion of apoceruloplasmin, copper-free and oxidase-inactive form, resulting from the disturbed incorporation of copper atoms into the protein (1, 14), while the intracellular synthesis of CPN...

Copper is an essential trace element in prokaryotes and eukaryotes and is strictly regulated by biological mechanisms. Menkes and Wilson diseases are human disorders that arise from disruption of the normal process of copper export from the cytosol to the extracellular environment. Recently a gene for Wilson disease (WD)(also named the ATP7B gene) was cloned. This gene encodes a copper transporter of the P-type ATPase. We prepared monoclonal and polyclonal anti-(WD protein) antibodies and characterized the full-length WD protein as well as a shorter form that is produced by alternative splicing in the human brain. We found that the WD protein is localized mainly in the Golgi apparatus, whereas the shorter form is present in the cytosol. These results suggest that the alternative WD proteins act as key regulators of copper metabolism, perhaps by performing distinct roles in the intracellular transport and export of copper.

Wilson's disease, an autosomal recessive disorder, is characterized by the excessive accumulation of hepatic copper that results from reduced biliary copper excretion and disturbed incorporation of copper into ceruloplasmin. The ATP7B gene, responsible for the disease, encodes a copper transporting P‐type ATPase. We previously demonstrated the involvement of ATP7B in hepatic copper secretion into plasma after the introduction of ATP7B into the Long‐Evans Cinnamon (LEC) rat, a rodent model of Wilson's disease. In this study we found the increased copper contents of the hepatic lysosomal fractions and bile in the LEC rats after ATP7B introduction, indicating the participation of ATP7B in the biliary excretory pathway for copper.

Matrine, also known as oxymatrine, is an important active ingredient of traditional Chinese herb Sophora flavescens. Recent studies have found that matrine may inhibit multiple tumors through inhibiting the tumor cell proliferation, inducing cell apoptosis, blocking cell cycle, suppressing cell invasion and migration and assisting in the synergy, and attenuation of radiotherapy and chemotherapy. This study mainly investigated the role of matrine in gastric cancer and its possible mechanism. The real‐time fluorescence quantitative polymerase chain reaction technique showed that matrine inhibited the proliferation and migration of gastric tumor cells and significantly suppressed the expression of miR‐93‐5p. The dual‐luciferase reporter gene assay indicated that AHNAK was a target gene of miR‐93‐5p and regulated by miR‐93‐5p and matrine. The torsion test demonstrated that matrine exerted its role via miR‐93‐5p while miR‐93‐5p played a role by targeting AHNAK. Thus, this study found that matrine affected the progression of gastric cancer by inhibiting the function of gastric cancer cells through the possible mechanism of inhibiting miR‐93‐5p expression to increase the expression level of the downstream target gene AHNAK.