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Objective To study the protein metabolism in rat myocardium after burn injury with sepsis, the mRNA expressions of ubiquitin were determined. Methods Wistar rats were subjected to a 30% full thickness burn and endotoxin (6mg/kg) was immediately injected into the peritoneal cavity. They were randomly divided into 2 and 6 hour groups and a normal control group, with 9 rats in each group. The cardiac muscle was taken to assay the mRNA expressions of ubiquitin during postinjury period. Results The expressions of the ubiquitin mRNA (2 4kb and 1 2kb), especially the 2 4kb stripe, in cardiac muscle of burn sepsis rats were significantly higher than that of normal control ( P
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To study the effects of TNF ? on the protein metabolism and the ubiquitin system gene expression in isolated skeletal muscles, after dissecting and isolating the extensor digitorium longus (EDL) muscles, the in vitro oxygen rich muscle incubation system and as high performance liquid chromatography were used to assess proteolytic rate of the samples. The EDL muscles in study group were incubated with media containing 6 000 U/ml recombinant rat TNF ?. In control group, the media were of the same composition as that of the study group except recombinant rat TNF ?. The expressions of ubiquitin mRNA and C 2 mRNA in rat EDL muscles were determined by Northern blot analysis. No notable difference was observed in the total and myofibrillar proteolytic rate in EDL muscles between the two groups. The expressions of ubiquitin mRNA(2 4kb) and C2 mRNA of EDL muscle incubated with medium containing TNF ? were increased by 151% and 56%, respectively, as compared with those in control group. TNF ? could directly strengthen the function of ubiquitin dependent proteolytic system, but further studies are necessay to elucidate whether TNF ? could directly increase the proteolytic rate in skeletal muscle.
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Objective: To investigate androgen receptor (AR) mRNA expression in human peripheral leukocytes. Methods: Total RNA was isolated from peripheral leukocytes of healthy men and women, AR mRNA was determined by RT PCR and Northern blot. Results: An AR cDNA fragment of the expected size of 390 bp was determined by RT PCR. 9.4 kb AR mRNA was detected by Northern blot, which was consistent with AR mRNA in human prostate reported by Lubahn et al . Conclusion:AR mRNA expresses in human peripheral leukocytes, this provides basis for studying the effect of androgen on the function of the human peripheral leukocytes and the changes of AR in pathological stages. [
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AIM: To confirm the expression level of the gene which corresponding to JT8 tag decreased in coronary artery disease (CAD). METHODS: The validity and reliability of the results gotten by serial analysis of gene expression method was verified by RT-PCR and Northern blot. The expression level of the gene which corresponding to JT8 tag was compared with the expression level of GAPDH and ?-actin in JT and WY, while according to SAGE results that the gene expression level of JT8 gene was 8 times higher in JT than in WY. RESULTS: It was found that the results of RT-PCR and Northern bolt were identified with the results of SAGE. The expression level of JT8 gene decreased in CAD. CONCLUSION: These results verified the validity of SAGE method and made a good foundation for further discovery of new candidate genes. [
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AIM: To confirm the differential expression genes in the rat ischemic brain. METHODS: The middle cerebral artery occlusion ischemic model was set up in rats. Fluorescence differential display reverse transcriptase-polymerase chain reaction (FDD RT-PCR) and reverse Northern blotting were used to fast confirm the differential expression genes. RESULTS: Nine differential expression sequence tags, including 6 known sequences and 3 unknown sequences, were confirmed. Among the known sequences, mus musculus ab1-interactor1,homo sapiens CGI-99 protein, tissue inhibitor of metalloproteinase 3 and homosapiens nuclear receptor co-repressor were up-regulated while homo-sapiens nuclear matrix protein p84 and coatomer protein complex, subunit gamma 2 were down-regulated. CONCLUSIONS: ① Combination of fluorescence differential display reverse transcriptase-polymerase chain reaction (FDD RT-PCR) and reverse Northern blotting is a method to fast-confirm the differential expression genes; ② There are differential expression genes in ischemic brain regions compared to non-ischemic parts. [
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Objective To study the expression and regulation of interleukin 15 (IL 15) in epidermal keratinocytes. Methods The expression of IL 15 mRNA in cultured normal human keratinocytes (NHKs) and human squamous cell carcinoma cell line (HSC 5) was analysed, and the effect of dexamethasone on IL 15 mRNA expression was studied by using RT PCR and Northern blotting technique. Results The results showed that both NHKs and HSC 5 expressed IL 15 constitutively. The level of IL 15 mRNA was significantly decreased after the cells were cultured with 10 -6 M dexamethasone. Conclusion It is suggested that keratinocyte derived IL 15 might be involved in the development of certain inflammatory skin diseases.
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AIM: To find out whether different dosage of rare earth element-lanthanum can influence the expression of aquaporin 7(AQP 7) in the testis of rats. METHODS:Rats were fed with lanthanum nitrate [La(NO 3) 3] and killed 6 months later. Testes were then removed immediately to extract total RNA. Northern blot analysis is performed finally. RESULTS:0.1 mg/kg La(NO 3) 3 depressed the expression of AQP 7 in rat testis, while 20 mg/kg La(NO 3) 3 had no significant effect on it. CONCLUSION: AQP 7 expession is found in the rat testis; La(NO 3) 3 can depress the expression of AQP 7 in the rat testis.
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Objective To study the subcellular localization and the tissue expression of EOLA1(endothelial-overexpressed lipopolysaccharide-associated factor 1). Methods The fusion protein EOLA1-EGFP expressed vector was constructed and transfected into endothelial cells. After 24 hours posttransfection, the subcellular localization of EOLA1 was detected by laser-scanning microscopy. The tissue-specific distribution of EOLA1 was assessed with Multiple Tissue Northern Blots. Results The expression of EOLA1 was tissue-specific in various human tissues. With human multiple tissue Northern blot analysis, it was shown that EOLA1 could express in the heart, skeletal muscle, kidney, liver, placenta, colon, spleen, small intestine, but did not in the brain, lung, thymus, and peripheral blood leukocyte. EOLA1 was mainly localized in cytoplasm and could move into the nucleus. Conclusion EOLA1 is one of intercellular proteins and may play a role in intercellular signal transduction.