Biomedical subjects

Fu-Chu He

Publications and source records attributed to Fu-Chu He.

23 records · Page 2Linked to original sources

PubMed2002

Identification and characterization of a novel isoform of hepatopoietin.

AIM: To isolate a novel isoform of human HPO (HPO-205) from human fetal liver Marathon-ready cDNA and characterize its primary biological function. METHODS: 5'-RACE (rapid amplification of cDNA 5' ends) was used to isolate a novel isoform of hHPO in this paper. The constructed pcDNA(HPO-205), pcDNA(HPO) and pcDNA eukaryotic expression vectors were respectively transfected by lipofectamine method and the stimulation of DNA synthesis was observed by (3)H-TdR incorporation assay. Proteins extracted from different cells were analyzed by Western blot. RESULTS: A novel isoform of hHPO (HPO-205) encoding a 205 amino acid ORF corresponding to a translated production of 23 kDa was isolated and distinguished from the previous HPO that lacked the N-terminal 80 amino acids. The dose-dependent stimulation of DNA synthesis of HepG2 hepatoma cells by HPO-205 demonstrated its similar biological activity with HPO in vitro. The level of MAPK (Mitogen-activated protein kinase) phosphorylation by Western blot analysis revealed that HPO-205 might have the stronger activity of stimulating hepatic cell proliferation than that of HPO. CONCLUSION: A novel isoform of hHPO (HPO-205) was isolated from hepatic-derived cells. The comparison of HPO-205 and HPO will lead to a new insight into the structure and function of hHPO, and provide the new way of thinking to deeply elucidate the biological roles of HPO/ALR.

Amino Acid Sequence↗

PubMed2002

[Isolation and characterization of a novel transcript of hepatopoietin].

A novel transcript of human HPO (HPO-205) encoding a 205 amino acid ORF corresponding to a translated product of 23 kD different from the previous HPO that lacks the N-terminal 80 amino acids, was isolated from human fetal liver cDNA library by 5'RACE methods. The dose-dependent stimulation of DNA synthesis of HepG2 hepatoma cells by HPO-205 demonstrated its similar biological activity with HPO in vitro. The level of MAPK (mitogen-activated protein kinase)phosphorylation was further examined by Western blot analysis and the result revealed that HPO-205 protein might have stronger activity on stimulating hepatic cell proliferation than that of HPO. The similar result was observed by FACS technique. Therefore, our results suggest that the HPO-205 has activity to stimulate proliferation of liver derived cells, and this activity may be stronger than HPO.

Amino Acid Sequence↗

PubMed2000

Development and Preliminary Application of a Peptide Mass Fingerprinting Technique in Proteome Research.

Proteome means the total proteins expressed by the genome in a cell or tissue. Two-dimensional electrophoresis (2-DE) is now the most powerful separating technique and the key separation method used in proteome. Peptide mass fingerprinting (PMF) is becoming a widely used and vastly developed technique for protein identification in 2-D gels. In this research, a systematic method to identify the proteins in polyacrylamide gels by PMF was developed. Proteins were digested in-gel by enzyme and the masses of generated peptides were measured by matrix assisted laser desorption ionization time of flight mass spectrometry (MALDI-TOF-MS). The data obtained from PMF were used in protein database search and the protein identification. The proteins from human lung cancer cells isolated by 2-DE were subjected to identification by the PMF method developed in this work. Three spots of proteins in gel were identified as G3P2_HUMAN, UBL1_ HUMAN and TPIS_HUMAN.

Journal Article↗

PubMed1998

Partial isolation and identification of hepatic stimulator substance mRNA extracted from human fetal liver.

AIM:To partially isolate and identify hepatic stimulator substance mRNA from human fetal liver tissues.METHODS:The poly (A)mRNA was extracted from human fetal liver tissues of 4-5 month gestation, fractionated by size on sucrose gradient centrifugation, translated into protein from each fraction in vitro and then its products were tested for HSS activity.RESULTS:Twenty-two 500 total RNA was obtained from human fetal liver tissues and pooled.mRNA of 420 was yielded,processed by oligo(dT)-cellulose column chromatography, then was size-fractionated by ultracentrifution on a continuous sucrose density gradient (5%-25%), and separated into 18 fractions.Translated products of mRNA in fraction 8 and 9 could produce a two-fold increase in the incorporation of( 3)H-TdR into DNA of SMMC-7721 hepatoma cells and in a heat resistant and organ-specific way.CONCLUSION:The partially purified HSS mRNA was obtained and this would facilitate the cloning of HSS using expression vectors.

Journal Article↗

PubMed1997

Expression and Activity of Recombinant Human Augmenter of Liver Regeneration.

The cDNA of human augmenter of liver regeneration was subcloned onto the downstream of the P(R)P(L) promoter of the expression plasmid pBV220. The recombinant plasmid could stably express ALR with high efficiency (up to 20% of the total bacterial proteins) in E. coli through thermal induction. The expressed recombinant ALRs could produce a significant increase in the incorporation of (3)H-TdR into liver DNA of a 1/3 hepatectomized test animal and also had a potent antihepatitis effect. These results suggest that ALR appears to be an important regulator of liver regeneration and may be used in clinical trial for enhancing liver regeneration in the treatment of hepatic diseases.

Journal Article↗