Experimental study on expression of ATP7B gene to restore copper transport in skin fibroblasts Me32aT22/2L cells / 中华神经医学杂志

Objective To observed hepatolenticular degeneration gene (ATP7B gene) expressed in Me32aT22/2L cell and detected whether A TP7B could transport redundant copper and reduce copper-induced cell apoptosis, which will make the basement for future gene therapy. Methods Piasmid pRc/CMV-WD containing A TP7B cDNA was transfected into Me32aT22/2L cell using lipofectamin2000 transfection methods. Intracellular distribution of ATP7B was detected by immunofluorescence histochemistry method and copper transportation function was studied using high-concentration copper incubation experiments, meanwhile, cell apoptosis induced by high-concentration copper incubation was observed. Results Expression of A TB7B gene could be detected and located around nuclei within the Me32aT22/2L cell. After incubation of high- concentration copper solution after 24, 48 and 72 hours, in empty vector group intracellular copper content was (600.60±69.71) ng/mg, (890.72±65.74) ng/mg and (1189.20±85.71) ng/mg respectively, however, intracellular copper content was (351.33±49.86) ng/mg, (427.38±30.95) ng/mg and (539.10±34.91) ng/mg in A TP7B group. Comparison of two groups has a statistical significance (P<0.01). Meanwhile, cell apoptotic rate was markedly decreased in A TPTB group compared with empty vector group (P<0.01). Conclusion Exogenous A TP7B gene could be expressed in Me32aT22/2L cell and A TPTB could transport intracellular redundant copper, subsequently reduced copper-induced cell apoptosis.

Study of copper metabolism and liver damage in TX Mice-an animal model for liver disease / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To provide right time points in selection of right aged animals and the normal physiological data of TX mice.</p><p><b>METHODS</b>7-12 months old TX and DL mice were studied, each group contained 3 female and 3 male mice of TX or DL mice. The concentration of copper in the serum, dry tissues (liver, brain and kidney), together with copper biochemistry indexes were measured. The liver histopathology was observed under light microscopy and electron microscope.</p><p><b>RESULTS</b>Transaminase increased significantly only in 10 and 11-month- old (AST(TX10) = 218.3 U/L, AST(TX11) = 197.5 U/L, AST(DL10) = 171.5 U/L, AST(DL11) = 165.0 U/L, P(10) less than 0.001, P(11) = 0.022), but the copper concentration of liver, brain and kidney was significantly increased during 7-12 month old (the average concentration of copper, Liver(TX) = (750.0 +/- 85.5) mg/kg, Brain(TX) = (39.7 +/- 2.2)mg/kg, Kidney(TX) = (29.8 +/- 5.0) mg/kg, Liver(DL) = (11.6 +/- 1.5) mg/kg, Brain(DL) = (16.8 +/- 0.9) mg/kg, Kidney(DL) = (14.2 +/- 1.0) mg/kg, t = 21.16, 23.60, 7.47, for all these organs P less than 0.05).</p><p><b>CONCLUSION</b>TX mice is a suitable model of liver disease with natural recovery, so selecting animal model of suitable time point is very important.</p>

Molecular genetics and its clinical application in the diagnosis of spinocerebellar ataxias / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To study the strategy of applying molecular genetic methods and techniques in the diagnosis of spinocerebellar ataxias (SCA).</p><p><b>METHODS</b>This study included 43 patients with SCA from 36 families, 38 sporadic SCA patients, 60 healthy individuals from the SCA families and 44 normal controls. The trinucleotide repeats were detected by polymerase chain reaction (PCR), denaturing polyacrylamide gel electrophoresis and silver staining technique. The repeat numbers were calculated by software.</p><p><b>RESULTS</b>SCA3 was the most common type in the Hans of south China, accounting for 42.0%, followed by SCA2 (7.4%), SCA1 (4.9%), SCA7 (3.7%), SCA6 (2.5%) and SCA12 (1.2%). No patient was found to have SCA8, SCA10, SCA17, and dentatorubro-pallidoluysian atrophy(DRPLA).</p><p><b>CONCLUSION</b>Molecular genetic detection is an effective way to confirmation of SCA subtype diagnosis and presymptomatic genetic diagnosis.</p>

Effect of the mutation of promoter region in Wilson disease ATP7B gene on the expression of reporter gene / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To find out the relationship between mutation of ATP7B gene promoter region and pathogenesis of Wilson disease(WD).</p><p><b>METHODS</b>Two of 48 WD patients presented C-->T base substitution mutations at the position -183. DNA sequences of the promoter region from normal and mutant samples were separated. The fragments containing the promoter region were cloned upstream of the luciferase. Luciferase activity was analyzed.</p><p><b>RESULTS</b>The luciferase activity of reporter gene containing normal sequence of ATP7B gene promoter region did not show significant difference as compared with that of reporter gene containing mutant promoter(n=3, P > 0.05).</p><p><b>CONCLUSION</b>No influence of C-->T base substitution mutations on the activity of promoter was observed in study. The results suggest that WD pathogenesis relates little to the mutations of the promoter region in Chinese.</p>

The expression of glomerulosclerosis by benazepril and its relationship with apoptosis of kidney cells / 实用儿科临床杂志

Objective To study the effects of angiotensin converting enzyme inhibitor benazepri1 on apoptosis and the expression of Fas and FasL in the kidney of rats with adriamycin-indued nephritic glomeruosclerosis.Methods After uninephrectomy and the injection of adriamycin induced rats model with glomerulosclerosis, benazapril(6 mg/kg) was delivered daily by gavage to the rats in therapeutic groups for 12 weeks.Apoptosis was examined by means of terminal-deoxynucleotidyl trans ferase mediated d-UTP nick end label ling(TUNEL) and immunohistochemistry was utlized to detect the expression of Fas and FasL.Software of pathological analysis quantitated the level of Fas and FasL.Results Compared with those of the control group, the kidney of model group had moresevere glomerulosclerosis, much more apoptotic cells and higher level of exprssion of Fas and FasL. The degree of glomeruloscleroais, the nuxner of apoptotic cells and the level of expression of Fas and FasL were ameliofated by benazepril treatment.Conclusion Benazepril may suppress the excessive apoptosis of kidney cell by lowering the expression of the protin correlatng apoptosis Fas and FasL,so as to postpone the process of glomeruosclerosis.