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Biomedical subjects

W Luo

Publications and source records attributed to W Luo.

At least 199 records · Page 11Linked to original sources

EGF abrogation-induced fusilli-form dysmorphogenesis of Meckel's cartilage during embryonic mouse mandibular morphogenesis in vitro.

Mutations associated with genes of the EGF superfamily are implicated in facial malformations arising from abnormal development of the first branchial arch. EGF and EGF receptor (EGFr) transcripts are expressed in the mouse embryonic first branchial arch and derivatives from E9 through E15. EGF transcripts are localized to ectomesenchymal cells associated with precartilage, cartilage, bone and tooth-forming cells. EGF and EGFr proteins co-localize to the same cells suggesting an autocrine regulation. To test whether EGF effects the timing and positional information required for Meckel's cartilage (MC) and tooth development, we cultured E10 mandibular explants in serumless, chemically defined medium with either antisense or sense EGF oligodeoxynucleotides. Antisense inhibition of EGF expression produces bilaterally symmetrical Fusilli-form dysmorphogenesis of MC and decreases tooth bud size; these effects are reversed by the addition of exogenous EGF to the culture medium. Tyrphostin RG 50864, which inhibits EGF receptor kinase activity, inhibits EGF stimulation of tyrosine phosphorylation in a concentration-dependent manner and severely retards mandibular development yet increases tooth size. These findings support the hypothesis that endogenous EGF and EGF-like proteins provide signalling to regulate the size and shape both of cartilage and tooth formation during craniofacial morphogenesis.

Animals↗

[A cephalometric study on determining the orientation of occlusal plane].

A study of the parallel relationship between the occlusal plane and the line connecting nasal alar and tragus was made in 90 dentulous cases by using cephalometry. The results show that the line connecting the inferior point of nasal alar and the mid-point of tragus runs much more parallel with the occlusal plane. The regression equation reveals a "line of closest fitting". It was used in the prosthetic treatment for 50 edentulous patients with good clinical results. The line connecting the inferior point of nasal alar and the mid-point of tragus therefore represents a proper reference plane for determining occlusal plane and hence should be still a valuable index in clinical dentistry.

Adolescent↗

[Computer analysis of evoked field potentials in hippocampal slice].

A microcomputer system for the processing of evoked field potentials of brain slices was developed on the IBM PC-AT microcomputer. In the system, a 385-point finite impulse response (FIR) filter was designed and fast Fourier convolution algorithm was adopted to attain both high quality of filtering and high efficiency of calculation. Also a method for wave peak recognition by analysing waveform slope was established and an interface circuit was designed for the automatic control of the FM tape recorder by the computer. As a result, automated parameter measurement of epileptiform activity recorded from hippocampal slices was successfully realized with high accuracy.

Animals↗

Rapid determination of blood serum retinol by reverse phase open column chromatography.

An aqueous, reverse phase, open column procedure (RP C18) was developed for the rapid determination of retinol in serum. The samples were extracted with hexane and separated on the column with methanol and water as the mobile phase. Two methods were developed depending on the size of the sample. For 2 ml of serum a spectrophotometer was used to quantify the sample and for 0.1 ml of serum a fluorometer was used. Phytofluene was retained on the top of the column and the column was reusable. The agreement between HPLC and the RP C18 open column was good. The lower limit of detection for micromethod was about 1.25 micrograms of retinol per dl of serum and the recovery of added retinol was quantitative.

Chromatography, High Pressure Liquid↗

Ly-1 (CD5), a membrane glycoprotein of mouse T lymphocytes and a subset of B cells, is a natural ligand of the B cell surface protein Lyb-2 (CD72).

CD5 is a 67-kDa glycoprotein expressed on the cell surface membrane of all T lymphocytes and on a small proportion of B lymphocytes. The physiologic role of this Ag is still unknown. Structural and functional studies of CD5 suggest that it might act as a receptor for a positive signal. CD5-specific mAb augment CD3- or mitogen-induced T cell proliferation, IL-2 secretion, and IL-2R expression and induce a rise in intracellular [Ca2+]. In this report, we describe the purification of mouse CD5 protein (mCD5) and its use as a probe to search for the ligand of CD5. We demonstrate that mCD5 specifically interacts with the mouse B cell differentiation Ag CD72/Lyb-2. Three serologically defined allelic forms of mouse CD72/Lyb-2 can all interact with mCD5. We further show that mCD5 can interact with human CD72/Lyb-2, and similarly, that human CD5 can interact with mouse CD72/Lyb-2. These studies may have major implications for the mechanisms of T-B cell communication.

Animals↗

Maintenance of amelogenin gene expression by transformed epithelial cells of mouse enamel organ.

Electroporation was used to introduce foreign genes into cells derived from the mouse enamel organ epithelia (EOE). Optimal conditions for this electroporation were established. The introduction of a plasmid construct bearing the coding region for the large T-antigen from polyoma virus into EOE cells permitted the establishment of a derivative cell line that has the following characteristics: (1) the cells could be passaged many times; (2) they expressed a keratin-containing cytoskeleton; and (3) approx. 60% of the cells expressed amelogenin, a tissue-specific gene product unique to ameloblasts. Potential uses for such a cell line include analysis of: (1) the upstream regulatory regions required for temporally and spatially restricted expression of amelogenin; (2) the post-translational modification of amelogenin in synchronized cells and (3) the organization and biomineralization of enamel extracellular matrix in monolayer culture.

Amelogenin↗

Human ameloblastoma tumors express the amelogenin gene.

Instructive signals are responsible for the regulation of the expression of gene products characteristic of many cell lineages during normal development and potentially during neoplasia. The odontogenic origin of ameloblastomas is based largely on the similarity in histologic appearance between the tumor and the developing tooth organ. A pathognomonic pattern for odontogenic tissue-specific gene expression in ameloblastomas has not been previously shown. In these studies, the gene expression parameters for human ameloblastomas have been characterized with the techniques of messenger RNA phenotyping in combination with Northern and in situ hybridization analysis of messenger RNA. The results of these studies confirm that amelogenin, a gene transcribed solely by differentiated ameloblasts, was expressed by epithelial cells from human ameloblastomas. This observation suggests that the instructive signals required for ameloblast differentiation are shared during normal development and tumorigenesis of odontogenic epithelium.

Ameloblastoma↗

Mouse phospholamban gene expression during development in vivo and in vitro.

To establish a murine model that may allow for definition of the precise role of phospholamban in myocardial contractility through selective perturbations in the phospholamban gene, we initiated studies on the role of phospholamban in the murine heart. Intact beating hearts were perfused in the absence or presence of isoproterenol, and quantitative measurements of cardiac performance were obtained. Isoproterenol stimulation was associated with increases in the affinity of the sarcoplasmic reticulum Ca2+ pump for Ca2+ that were due to phospholamban phosphorylation. To assess the regulation of phospholamban gene expression during murine development, Northern blot and polymerase chain reaction analyses were used. Phospholamban mRNA was first detected in murine embryos on the ninth day of development (the time when the cardiac tube begins to contract). In murine embryoid bodies, which have been shown to recapitulate several aspects of cardiogenesis, phospholamban mRNA was detected on the seventh day (the time when spontaneous contractions are first observed). Only those embryoid bodies that exhibited contractions expressed phospholamban transcripts, and these were accompanied by expression of the protein, as revealed by immunofluorescence microscopy. Sequence analysis of the cDNA encoding phospholamban in embryoid bodies indicated complete homology to that in adult hearts. The deduced amino acid sequence of murine phospholamban was identical to rabbit cardiac phospholamban but different from dog cardiac and human cardiac phospholamban by one amino acid. These data suggest that phospholamban, the regulator of the Ca(2+)-ATPase in cardiac sarcoplasmic reticulum, is present very early in murine cardiogenesis in utero and in vitro, and this may constitute an important determinant for proper development of myocardial contractility.

Amino Acid Sequence↗

Epidermal growth factor transcription, translation, and signal transduction by rat type II pneumocytes in culture.

Epidermal growth factor (EGF) is known to induce fetal lung maturation and its receptor is present in the lungs of several species. Recently, EGF has been immunolocalized in type II pneumocytes in rat lung. We postulated that EGF is synthesized in type II pneumocytes and that, because of its position-restricted distribution within the alveolus, EGF might act as an autocrine regulator of type II pneumocyte function. Herein, we have tested the hypothesis using adult rat type II pneumocytes in primary culture. In situ hybridization, using an oligonucleotide probe corresponding to amino acid residues 1070 to 1081 of mouse EGF precursor, demonstrated the presence of EGF precursor mRNA. Upon S-200 Sephacryl gel chromatography of type II pneumocyte extracts, EGF-reactive protein eluted as a high-molecular-weight form (greater than 100 kD). EGF immunoreactivity was localized within type II pneumocytes in the periphery of groups of 10 to 15 cells in culture. The type II pneumocytes bound [125I]EGF in a specific manner, indicating the presence of EGF receptors. Scatchard plots gave an apparent affinity constant (Ka) of 1 x 10(9) liters/mol, and the number of receptors was estimated to be 4.8 x 10(11) mg protein (50 per cell). EGF receptor binding specificity was confirmed by the absence of an autoradiographic signal for cells incubated in the presence of a 100-fold excess concentration of transforming growth factor-alpha. Binding of [125I]EGF could also be downregulated 95% by incubation with 0.2 nM transforming growth factor-alpha.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

The protective effect of colchicine on bleomycin-induced pulmonary fibrosis in rats.

With the purpose of evaluating the therapeutic effect of colchicine on lung fibrosis in rats, bleomycin A5 (BLM-A5) was injected intratracheally to produce a lung fibrosis model. The animals were then treated with colchicine, 50 micrograms/d i.m. One month later, collagen fiber deposition scores (stained by Masson trichrome) were significantly lower in the treated group than in the untreated group (P less than 0.01). This result was proved further by assaying total lung hydroxyproline content (P less than 0.05). Histopathologic findings showed that the proliferation of fibroblasts dropped slightly in the treated group as compared with the untreated group. From this study, we conclude that colchicine has certain antifibrotic effects and may be used in the treatment of pulmonary fibrosis.

Animals↗

[Epidemiological study of children's mental health in Fuzhou].

This epidemiological survey of mental health's conditions among 1,428 children below the age of 14 in an urban and rural area with a population of 398,674 was carried out with the standardized methods and diagnostic criteria, the result shows that the total incidence of abnormal mental health's conditions is 70.03/1000. Through a statistical deal, the result shows that the weights of these infants are slight, that they can only recognize their parents slowly, that their parents are in low educated levels. That the emotion between their parents is always not so harmonious, and that their parents always treat their children with different attitudes. In short, all these factors are in close connection with the incidence of abnormal mental health of these children.

Adolescent↗

The B-cell surface protein CD72/Lyb-2 is the ligand for CD5.

The glycoprotein CD5 is expressed on the surface membrane of all mature T cells and a small proportion of B lymphocytes. Its exact role in immune interactions is still unknown. Studies indicate that CD5 functions both in mice and humans as a receptor, delivering co-stimulatory signals to T cells in a manner similar to CD2 (ref. 11) and CD28 (ref. 12). Anti-CD5 antibodies stimulate both T-cell proliferation mediated by CD3 in association with the T-cell receptor and secretion of interleukin-2 and expression of its receptor, as well as inducing an increase in intracellular Ca2+ concentration (refs 5-10). To identify the ligand for CD5 we purified the human CD5 protein, labelled it with biotin and used it as a probe. Here we report that CD5 specifically interacts with the cell-surface protein CD72 exclusive to B cells. This interaction is blocked by anti-CD72 antibodies, but not by any other anti-B-cell antibodies. Moreover, non-B cells (mouse L-cell fibroblasts and human Jurkat T cells) expressing a transfected human CD72 complementary DNA could bind to the CD5-biotin conjugate. The results demonstrate that the B-cell surface protein CD72 (Lyb-2 in mice) is the ligand for CD5.

Antigens, CD↗

Human interleukin-9: genomic sequence, chromosomal location, and sequences essential for its expression in human T-cell leukemia virus (HTLV)-I-transformed human T cells.

We have isolated the genomic sequence of human interleukin-9 (IL-9) based on its sequence homology with a human IL-9 cDNA isolated from human T-cell leukemia virus (HTLV)-I-transformed T cells by expression cloning. The entire genomic sequence has been determined and the gene consists of five exons and four introns. The human IL-9 gene is mapped to the long arm of human chromosome 5 at band 5q31-32, a region found to be deleted in a number of patients with acquired 5q- abnormalities and hematologic disorders. Several blocks of transcriptional control sequences have been identified at the 5'-flanking region of the human IL-9 gene that may play an important role in the control of IL-9 gene expression. The 5'-regulatory region of the human IL-9 gene also contains sequences identified in the 5'-flanking regions of other cytokine genes mapped to the long arm of human chromosome 5, including IL-3, IL-4, IL-5, and granulocyte-macrophage colony-stimulating factor and other T-cell growth factor genes including IL-2 and IL-6. The IL-9 gene is constitutively expressed in the HTLV-I-transformed human T cells and the expression of IL-9 in these cells can be further induced by 12-O-tetradecanoyl phorbol 13-acetate. Transient transfection analysis using the plasmid containing the 5'-flanking region of IL-9 gene upstream from the firefly luciferase ciferase report gene indicated that the 0.9-kb Smal-Sacl fragment of the IL-9 gene contains sequences required for the constitutive and activated expression of IL-9 gene in HTLV-I-transformed cells. These results will now allow us to study the regulatory mechanism of IL-9 gene expression in normal and leukemic human T cells.

Amino Acid Sequence↗

Cells from Hertwig's epithelial root sheath do not transcribe amelogenin.

Recent experimental evidence has led to the interpretation that "enamel-like" material is deposited along the forming mouse molar root surface by cells of Hertwig's epithelial root sheath (HERS cells) and that this material is integral to the developmental program for cementogenesis. The experimental strategy described in this study was to examine selected developmental stages of root formation for mouse first and second mandibular molars in order to localize the cellular sites of amelogenin gene transcripts using high resolution in situ hybridization. Amelogenin is the major structural protein of coronal enamel and is highly conserved among mammalian species at the DNA and amino acid sequence level. Within the limits of sensitivity for in situ hybridization and utilizing either cRNAs or oligodeoxynucleotide probes, we were unable to localize amelogenin transcripts within HERS cells from selected developmental stages associated with mouse molar root formation. In contrast, previous studies using antipeptide antibodies have provided immuno-histochemical localization of amelogenin domains in HERS cell-derived products. For these HERS cell-derived proteins to contain both amelogenin epitopes and yet fail to yield nucleic acid hybridization signals suggests that either gene rearrangement and/or alternative processing of messenger RNAs from the structural gene locus operate to produce immunologically related motifs sharing insufficient complementarity at the nucleotide level to permit efficient detection by hybridization. It is postulated that HERS cells synthesize proteins which contain amelogenin domains and that these proteins participate during cementogenesis. However, these enamel-related proteins are neither identical to, nor collinear with coronal canonical amelogenin transcripts.

Ameloblasts↗

[Pulmonary infections in renal transplant recipients].

Fifty-seven episodes of pulmonary infection occurred in 46 patients during a 11.5-year period, the incidence being 12% (57/475). After treatment, 22 patients recovered, and 24 died (52%). The etiology was determined in 47 infection episodes. 38(81%) of them were caused by bacteria, 6 by fungi, and 3 by tubercle bacilli. Comparisons were made between infected and noninfected groups. The results showed that the incidence of rejection episodes, greatly elevated blood sugar level (greater than or equal to 11.1 mmol/L) and granulocytopenia (less than or equal to 1.5 10(9)/L) influenced significantly the morbidity and mortality of pulmonary infection.

Adolescent↗