Search PubMed⌕ Search

Biomedical subjects

H Cao

Publications and source records attributed to H Cao.

At least 217 records · Page 12Linked to original sources

Genomic organization and tissue expression of the mouse proteasome gene Lmp-7.

LMP7 is one of the two proteasome subunits encoded in the major histocompatibility complex and is speculated to play a role in the generation of endogenous peptides for presentation by class I molecules to cytotoxic T cells. Here we report the genomic organization of the mouse Lmp-7 gene and the tissue distribution of its messenger RNA. In contrast to human LMP7 which is composed of seven exons and six introns, the mouse Lmp-7 gene is organized in six exons and five introns. Interestingly, the region corresponding to the first exon of human LMP7 is highly modified by numerous insertions and deletions and contains two in frame stop codons. Consequently, the mouse Lmp-7 gene does not allow the alternative exon usage described in humans and most likely encodes for only one LMP7 protein. Thus, the Tap-1 3' end gene region and the Lmp-7 initial translation codon are separated by an 1182 nucleotide region which contains a TATA-box, a cAMP regulatory element, two SP1 sites, and two G-C-rich regions. Expression of the Lmp-7 messenger RNA was analyzed on different tissues from unstimulated mice. Lmp-7 messenger RNA is expressed in spleen, thymus, lung, liver, heart, and, at a very low level, in kidney but not in brain and testis. The possible role of Lmp genes in antigen processing is discussed.

ATP Binding Cassette Transporter, Subfamily B, Mem↗

Novel self-regulation of human chorionic gonadotropin biosynthesis in term pregnancy human placenta.

Term pregnancy human placenta contains hCG/LH receptor mRNA transcripts and immunoreactive receptor protein. Both the receptor transcripts and receptor proteins are present only in trophoblasts. These findings led us to investigate whether hCG can regulate its own synthesis in term pregnancy human placenta. Treatment of placental tissue in static cultures or in a dynamic superfusion system with increasing concentrations of highly purified hCG provoked a biphasic effect on the steady state hCG subunit mRNA levels. Although low concentrations of hCG (< 200 mIU/ml) were not effective, moderate concentrations (200-1000 mIU/ml) increased, and high concentrations (> or = 5000 mIU/ml) either had no effect or actually decreased mRNA levels relative to the control values. This response was specific, because none of the hCG concentrations tested had any effect on glyceraldehyde-3-phosphate dehydrogenase or beta-actin mRNA levels. The effects of hCG on steady state hCG subunit mRNA levels were paralleled by corresponding changes in tissue hCG protein levels. Endogenous hCG appears to down-regulate alpha-subunit mRNA levels and hCG secretion. The hCG effect is probably receptor mediated, because a receptor antagonist, deglycosylated hCG, partially antagonized the hCG action. Treatment with exogenous hCG also down-regulated its own receptor mRNA and receptor protein levels. hCG regulation of its alpha-subunit and receptor levels involved both transcriptional as well as posttranscriptional mechanisms. In summary, this is the first demonstration of hCG regulating its own synthesis in term pregnancy human placenta. The findings of this study could offer a potential molecular explanation for the profile of hCG levels in normal pregnant women.

Antibodies↗

[Intracerebral-ventricular injection of 4-aminopyridine induced convulsion in rabbits].

Intracerebral-ventricular (icv) injection of 4-aminopyridine (4-AP) 8 micrograms to rabbits generated convulsions and epileptic discharges in electrocorticogram (ECoG). With 40 rabbits, the occurrence of convulsion at this dose level was 100%. The results from five rabbits showed that seizures may break out repeatedly at intervals of 1-5 min and such state may last about 1.5 h. Antiepileptic drugs, such as phenytoin sodium, phenobarbital sodium and diazepam, were found to effectively control the seizures provoked by icv 4-AP. Sodium valproate was less effective than the foregoing drugs. The icv 4-AP-induced convulsion appeared to have some merits as compared with the old epilepsy models, such as those induced by electric stimulation, cardiazol, or icv ferrous sulfate. We feel that it may be recommended as a model for the preliminary screening of antiepileptic drugs. The seizures provoked by icv 4-AP were shown to be antagonized by scopolamine, haloperidol, phentolamine, or propranolol, but potentiated by l-dopa. It is now recognized that 4-AP can enhance the release of ACh, DA, and NE in the central nervous system. The present results suggest that the convulsion elicited by 4-AP seems to be related to the disturbance of synaptic transmission in the brain.

4-Aminopyridine↗

Evidence of the specific molecular interactions between naringenin, NODD and nod-promoter.

This paper presents evidence that specific molecular interactions occur between three nod regulation elements: naringenin, NODD and nod-promoter. No such interactions were observed when the concentration of naringenin was less than 0.4 mM. However, remarkable molecular interactions were observed when naringenin was at 4.0 mM. The interactions were highly specific because no such interactions have been observed when naringenin was exposed at various concentrations to other promoter-regulatory protein system (e.g., the protease regulatory system of Xanthomonas campestris, although, like NODD-nod-promoter system, specific binding occurs between the protease promoter and the corresponding regulatory protein).

Endopeptidases↗

[Determination of aconitine in wutou and related herbal drugs (roots of Aconitum) and their processed products in Hong Kong market].

This paper deals with the determination of the poisonous aconitine-type alkaloids in roots of Aconitum in Hong Kong market by TLC scanning. The results show that the content of aconitine was 0.0041%-0.021% in processed roots of Aconitum carmichaeli, 0.0084%-0.034% in processed roots of A. kusnezoffii; 0.0012% and 0.0007% in crude and roasted lateral tuber of A. carmichaeli. Aconitine was not found in the processed product of Fuzi.

Aconitine↗

An extra cysteine proximal to the transmembrane domain induces differential cross-linking of p185neu and p185neu.

The neu proto-oncogene encodes a receptor tyrosine kinase (p185) that is closely related to the epidermal growth factor receptor. It has been proposed that receptor tyrosine kinases are activated through oligomerization. Because this clustering model predicts that oligomerization of receptors is sufficient to activate them, we determined if p185 can be activated by introducing an extra cysteine proximal to the transmembrane domain. This should induce inter-receptor disulfide bonding and, according to the clustering model, activate the receptor. This amino acid substitution enhanced recovery of both normal and transforming neu proteins as dimers, with normal p185 recovered predominantly as monomers and transforming p185* as dimers. However, the cysteine substitution did not affect the transforming activity of the two proteins.

3T3 Cells↗

A subdomain in the transmembrane domain is necessary for p185neu* activation.

The neu proto-oncogene encodes a protein highly homologous to the epidermal growth factor receptor. The neu protein (p185) has a molecular weight of 185,000 Daltons and, like the EGF receptor, possesses tyrosine kinase activity. neu is activated in chemically induced rat neuro/glioblastomas by substitution of valine 664 with glutamic acid within the transmembrane domain. The activated neu* protein (p185*) has an elevated tyrosine kinase activity and a higher propensity to dimerize, but the mechanism of this activation is still unknown. We have used site-directed mutagenesis to explore the role of specific amino acids within the transmembrane domain in this activation. We found that the lateral position and rotational orientation of the glutamic acid in the transmembrane domain does not correlate with transformation. However, the primary structure in the vicinity of Glu664 plays a significant role in this activation. Our results suggest that the Glu664 activation involves highly specific interactions in the transmembrane domain of p185.

3T3 Cells↗

China's Cooperative Medical System: its historical transformations and the trend of development.

The rural Cooperative Medical System (CMS) in China came into existence in the 1950s. Along with the development of the collective economy of agriculture and the promotion of this system by the Chinese Government, the coverage rate of the CMS in the rural areas increased from 1955 through the end of the 1970s, when it was estimated that 90 percent of the rural population was covered by the CMS. Since the rural economic reform in the early 1980s destroyed the rural collective economy on which the CMS was based, The coverage rate of the CMS went down to 5 percent. After 1985, the coverage rate of the CMS began to increase gradually because of the development of the collective fund for supporting the CMS and the governments' efforts to strengthen the CMS. The structure of CMS has changed over time. Traditionally, the CMS was organized at the village level. In order to increase the risk-sharing ability of this system, the trend is for it to be organized at the township level in the future.

China↗

[Immortalization of cultured cells by the Ha-rasEJ oncogene and expression of a gene of biogenetic value].

The double recombination into NIH-3T3 cells of the cloned Ha-rasEJ oncogene and the cDNA gene of the human growth hormone (hGH) under the control of a heat inducible promoter (hsp70) allowed hGH production either in vitro using the mass culture of engineered cells in biogenerators, or in vivo after xeno-transplantation of the cells into an animal host. Therefore the in vivo synthesized hGH induced the production of anti-hGH polyclonal antibodies.

3T3 Cells↗

[Identification of seven snake-drugs by iso-electric point].

This paper deals with the identification of seven snake-drugs by means of iso-electric point of protein. The experimental results show that different species of snake-drugs have different iso-electric points. The experimental data are accurate and easily reproducible (CV less than 0.5%). The difference of the averages of iso-electric points of the seven snake-drugs (P less than 0.01%) is extremely significant and thus provides a scientific criterion for the identification of peeled or powdered snake-drugs.

Animals↗

TPA inhibits the tyrosine kinase activity of the neu protein in vivo and in vitro.

P185 is a receptor-like protein encoded by the neu/erbB-2 proto-oncogene. A point mutation in the transmembrane domain renders this protein oncogenic. We report here that incubation of cells with 12-O-tetradecanoylphorbol-13-acetate (TPA) stimulates the phosphorylation of the normal neu protein (p185) and the oncogenic neu protein (p185*). The increased phosphorylation occurs mainly on serine and threonine residues. Phosphate labeling experiments showed that TPA causes a reduction of basal phosphotyrosine in p185 but not p185*. Immunoblotting with antiphosphotyrosine antibody yielded similar results. TPA also inhibited tyrosine phosphorylation of p185* in an in vitro immune complex kinase assay. These data suggest that protein kinase C, the receptor for TPA, regulates p185 function through serine or threonine phosphorylation.

Amino Acids↗

HDV (delta factor) infection complicated with subacute fulminant hepatitis B. A morphological and immunohistochemical observation.

In 75 patients with subacute fulminant hepatitis B (SAFH), HDAg positive liver cells were observed in 14 cases (18.67%) by direct enzyme labelled method. It was found further that HDV (delta factor) infection was an important cause in producing massive necrosis of liver cells and superinfection of HDV with hepatitis B was identified as one of the causes of SAFH. This experiment also showed that the number of HDAg positive cells was in direct proportion to the area of liver necrosis. No obvious lymphocytic infiltrations and cytotoxic phenomena around the necrotic foci and HDAg positive cells could be found, while the cytoplasmic type HDAg positive cells showed distinct degeneration or even atrophy. Hence it was suggested that HDV could be a direct pathogenesis. No obvious characteristic changes in morphology of HDV infection could be found.

Antigens, Viral↗

Differential regulation of class II MHC determinants on macrophages by IFN-gamma and IL-4.

Initiation of an immune response depends upon expression of class II MHC determinants on plasma membranes of APC. Murine peritoneal macrophages treated with either rIFN-gamma or rIL-4 display significantly more class II MHC determinants than untreated control cells. Analysis of the induction of macrophage Ia Ag by these cytokines showed considerable quantitative and qualitative differences. Maximal levels of Ia Ag induced in macrophages and detected by ELISA after IL-4 treatment at 48 h was about 80% of that induced by IFN-gamma. However, the frequency of Ia+ cells in replicate macrophage populations cultured for 48 h in excess concentrations of cytokine was 60 to 80% with IFN-gamma, 30 to 40% with IL-4, and 5% with medium alone. Thus, the subpopulation of macrophages able to respond to IL-4 for induction of Ia Ag expression was less than that able to respond to IFN-gamma. Expression of Ia Ag on macrophages continuously exposed to IFN-gamma was maximal at 48 h and remained at this high level through 6 days. Maximal Ia Ag expression for IL-4-treated cells was also detected at 48 h, but was not sustained with time in culture, and returned to base line by 4 days. A similar time course for levels of Ia-specific message in macrophages at various times after IFN-gamma and IL-4 treatment was detected by Northern dot blot analysis. Loss of Ia mRNA and Ag with time in culture in the IL-4 treated cells was not due to macrophage cell death, depletion of active cytokine, or presence of fluid-phase inhibitors. IL-4 unresponsive cells were fully capable of maximal response to IFN-gamma for Ia Ag induction. These findings suggest that IL-4 and IFN-gamma induce class II MHC determinants through different mechanisms which may provide discrete regulatory control of APC function.

Animals↗

Cell-surface expression of a membrane-anchored form of the human chorionic gonadotropin alpha subunit.

We carried out experiments designed to generate a novel cell-surface protein from a small glycosylated secretory protein. DNA encoding the entire precursor of human chorionic gonadotropin (hCG, alpha subunit) was fused precisely to DNA encoding the transmembrane and cytoplasmic domains of the vesicular stomatitis virus glycoprotein. When expressed in animal cells this DNA encoded the 92-amino acid hCG-alpha subunit anchored in cellular membranes by an extension composed of the 49 carboxyl-terminal amino acids of vesicular stomatitis virus glycoprotein. This hybrid protein was transported efficiently to the plasma membrane of animal cells. The two asparagine-linked glycans on the anchored form of hCG-alpha were large and heterogeneous when compared to those on the secretory form. Experiments employing in vitro mutagenesis and the glycosylation inhibitor tunicamycin established that the presence of at least one of the two asparagine-linked glycans was required for expression of the anchored molecule on the cell surface. However, as reported previously, secretion of hCG-alpha occurred in the absence of glycosylation. Also, mutations eliminating the second glycosylation site (at amino acid 78) in both the anchored or secreted forms apparently led to partial denaturation or a conformational change interfering with transport of the protein.

Biological Transport, Active↗

Effects of altered cytoplasmic domains on transport of the vesicular stomatitis virus glycoprotein are transferable to other proteins.

Alterations of the cytoplasmic domain of the vesicular stomatitis virus glycoprotein (G protein) were shown previously to affect transport of the protein from the endoplasmic reticulum, and recent studies have shown that this occurs without detectable effects on G protein folding and trimerization (R. W. Doms et al., J. Cell Biol., in press). Deletions within this domain slowed exit of the mutant proteins from the endoplasmic reticulum, and replacement of this domain with a foreign 12-amino-acid sequence blocked all transport out of the endoplasmic reticulum. To extend these studies, we determined whether such effects of cytoplasmic domain changes were transferable to other proteins. Three different assays showed that the effects of the mutations on transport of two membrane-anchored secretory proteins were the same as those observed with vesicular stomatitis virus G protein. In addition, possible effects on oligomerization were examined for both transported and nontransported forms of membrane-anchored human chorionic gonadotropin-alpha. These membrane-anchored forms, like the nonanchored human chorionic gonadotropin-alpha, had sedimentation coefficients consistent with a monomeric structure. Taken together, our results provide strong evidence that these cytoplasmic mutations affect transport by affecting interactions at or near the cytoplasmic side of the membrane.

Autoradiography↗