Search PubMed⌕ Search

Biomedical subjects

X Cao

Publications and source records attributed to X Cao.

At least 91 records · Page 5Linked to original sources
PubMed2001

Clinical observation on preservation of lens anterior capsule in reduction of complications associated with silicone oil.

PURPOSE: To observe the effect of preservation of anterior capsule on the incidence of complications associated with silicone oil. METHODS: 82 patients (82 eyes) accepted trans pars plana vitrectomy (TPPV) combined with lensectomy of whom, 30 eyes with preservation of anterior capsule (PAC) and 52 eyes with no preservation of anterior capsule (NPAC). Analysis of the incidence of complications associated with use of oil. RESULTS: The incidence rate was 50.0% in NPAC group, and 23.3% in PAC group (P < 0.025). There were secondary glaucoma (21.1%), band keratopathy (13.5%) and corneal decompensation (9.6%) in NPAC group, while there was none of them in PAC group. CONCLUSIONS: Preservation of anterior capsule is an effective method to reduce the complications associated with silicone oil.

Adolescent↗
PubMed2001

[Raman spectra of Cu/Zn superoxide dismutase and L-cysteine used as an electron transfer promoter].

In author's laboratory, it has been demonstrated with electrochemical techniques that L-cysteine can be used as an effective electron transfer promoter to accelerate the electron transfer process of Cu/Zn superoxide dismutase on gold electrode. In order to reveal the mechanism of this promotion, confocal microscopic Raman spectroscopic technique is employed here to obtain Raman spectra of solid state SOD and L-cysteine modified gold electrode. Moreover, Raman spectra of L-cysteine modified gold electrode with two different approaches are also obtained. Analysis and discussion for these Raman spectra are reported.

Cysteine↗
PubMed2001

[Color reaction of chromogenic reagent O-hydroxybenzenediazoaminoazobenzene with thallium (III) and its application].

The chromogenic reaction of the O-hydroxybenzenediazoaminoazobenzene (O-HDAA) with thallium was studied in this paper. In the presence of Triton X-100 and SDBS and in ammonia medium of 0.54-1.1 mol.L-1, O-HDAA reacts with thallium (III) to form a stable red complex with molar radio of 1:5. The apparent molar absorptivity is 1.4 x 10(5) L.mol-1.cm-1 at the maximum absorption wavelength of 520 nm. Beer's law is obeyed in the concentration range of 0-600 micrograms.L-1 Tl(III). The method has been applied to the direct determination of trace amounts of thallium in waste water and in geological samples by using polyurethane foam to absorb and separate Tl(III) with satisfactory results.

Color↗
PubMed2000

Identification and characterization of a novel gene KE04 differentially expressed by activated human dendritic cells.

To better understand the molecular mechanisms of dendritic cells (DC) function, we isolated differentially expressed genes in Ag-activated DC by a PCR-based subtractive hybridization technique. A novel full-length cDNA, KE04, was thus isolated from KLH-activated human PBMC-derived DC. KE04 cDNA encoded a 346-aa protein devoid of functionally indicative motifs. KE04 protein showed 64% identity with a Caenorhabditis elegans protein and 83% identity with a human putative protein. Distant relationship was also found with other prokaryotic and eukaryotic proteins. Differential expression of KE04 in activated DC other than nonactivated DC was confirmed at both mRNA and protein levels. KE04 mRNA expression was detectable in various human tissues and cell lines by Northern blot and RT-PCR. Western blot and confocal microscopy analysis indicated that its cytolocalization was intracellular. Our data suggest the potential involvement of KE04 in DC activation and will facilitate the research of molecular mechanism of DC function.

Amino Acid Sequence↗
PubMed2000

13C/31P NMR assessment of mitochondrial energy coupling in skeletal muscle of awake fed and fasted rats. Relationship with uncoupling protein 3 expression.

To examine the relationship between mitochondrial energy coupling in skeletal muscle and change in uncoupling protein 3 (UCP3) expression during the transition from the fed to fasted state, we used a novel noninvasive (31)P/(13)C NMR spectroscopic approach to measure the degree of mitochondrial energy coupling in the hind limb muscles of awake rats before and after a 48-h fast. Compared with fed levels, UCP3 mRNA and protein levels in the gastrocnemius increased 1.7- (p < 0.01) and 2.9-fold (p < 0.001), respectively, following a 48-h fast. Tricarboxylic acid cycle flux measured using (13)C NMR as an index of mitochondrial substrate oxidation was 212 +/- 23 and 173 +/- 25 nmol/g/min (p not significant) in the fed and 48-h fasted groups, respectively. Unidirectional ATP synthesis flux measured using (31)P NMR was 79 +/- 15 and 57 +/- 9 nmol/g/s (p not significant) in the fed and 48-h fasted groups, respectively. Mitochondrial energy coupling as expressed by the ratio of ATP synthesis to tricarboxylic acid cycle flux was not different between the fed and fasted states. To test the hypothesis that UCP3 may be involved in the translocation of long chain free fatty acids (FFA) into the mitochondrial matrix under conditions of elevated FFA availability, [U-(13)C]palmitate/albumin was administered in a separate group of rats with (+) or without (-) etomoxir (an inhibitor of carnitine palmitoyltransferase I). The ratio of glutamate enrichment ((+) etomoxir/(-) etomoxir) in the hind limb muscles was the same between groups, indicating that UCP3 does not appear to function as a translocator for long chain FFA in skeletal muscle following a 48-h fast. In summary, these data demonstrate that despite a 2-3-fold increase in UCP3 mRNA and protein expression in skeletal muscle during the transition from the fed to fasted state, mitochondrial energy coupling does not change. Furthermore, UCP3 does not appear to have a major role in FFA translocation into the mitochondria. The physiological role of UCP3 following a 48-h fast in skeletal muscle remains to be elucidated.

Adenosine Triphosphate↗
PubMed2000

Evidence of the synthesis of opioid receptor like 1 receptor in nociceptinergic neurons in rat brain suggests the existence of autoreceptor: a confocal double staining study.

Opioid receptor like 1 (ORL1) receptor is a novel member of the opioid receptor family, which was not bound by any of the typical opioid receptor ligands but bound by the recently discovered nociceptin (also termed orphanin FQ) with high affinity. By using double staining of fluorescent in situ hybridization and immunohistochemistry, we observed the expression of ORL1 receptor mRNA in nociceptin-like immunoreactive neurons in multiple areas in rat brain including the hippocampus, arcuate nucleus in the hypothalamus, ventralateral periaquiductal gray (PAG) and raphe nuclei in brain stem. The expression of ORL1 mRNA in nociceptinergic neurons suggests that these receptors mediate, at least in part, the presynaptic autoreceptor functions. Further anatomical and functional significance of the autoreceptor of nociceptinergic neurons remain to be elucidated.

Animals↗
PubMed2000

Molecular cloning and characterization of a novel CXC chemokine macrophage inflammatory protein-2 gamma chemoattractant for human neutrophils and dendritic cells.

Chemokines play important roles in leukocyte trafficking as well as function regulation. In this study, we described the identification and characterization of a novel CXC chemokine from a human dendritic cell (DC) cDNA library, the full-length cDNA of which contains an open reading frame encoding 111 aa with a putative signal peptide of 34 aa. This CXC chemokine shares greatest homology with macrophage inflammatory protein (MIP)-2alphabeta, hence is designated as MIP-2gamma. Mouse MIP-2gamma was identified by electrocloning and is highly homologous to human MIP-2gamma. Northern blotting revealed that MIP-2gamma was constitutively and widely expressed in most normal tissues with the greatest expression in kidney, but undetectable in most tumor cell lines except THP-1 cells. In situ hybridization analysis demonstrated that MIP-2gamma was mainly expressed by the epithelium of tubules in the kidney and hepatocytes in the liver. Although no detectable expression was observed in freshly isolated or PMA-treated monocytes, RT-PCR analysis revealed MIP-2gamma expression by monocyte-derived DC. Recombinant MIP-2gamma from 293 cells is about 9.5 kDa in size and specifically detectable by its polyclonal Ab developed by the immunization with its 6His-tagged fusion protein. The eukaryotically expressed MIP-2gamma is a potent chemoattractant for neutrophils, and weaker for DC, but inactive to monocytes, NK cells, and T and B lymphocytes. Receptor binding assays showed that MIP-2gamma does not bind to CXCR2. This implies that DC might contribute to the innate immunity through the production of neutrophil-attracting chemokines and extends the knowledge about the regulation of DC migration.

Adult↗
PubMed2000

Cytochrome c release and apoptosis induced by mitochondrial targeting of nuclear orphan receptor TR3.

TR3, an immediate-early response gene and an orphan member of the steroid-thyroid hormone-retinoid receptor superfamily of transcription factors, regulates apoptosis through an unknown mechanism. In response to apoptotic stimuli, TR3 translocates from the nucleus to mitochondria to induce cytochrome c release and apoptosis. Mitochondrial targeting of TR3, but not its DNA binding and transactivation, is essential for its proapoptotic effect. Our results reveal a mechanism by which a nuclear transcription factor translocates to mitochondria to initiate apoptosis.

Apoptosis↗
PubMed2000

Cloning of DPK, a novel dendritic cell-derived protein kinase activating the ERK1/ERK2 and JNK/SAPK pathways.

Mitogen-activated protein kinase (MAPK) cascades are the major signaling systems transducing extracellular signals into intracellular responses, which mainly include the extracellular signal-regulated kinase (ERK) pathway, the c-Jun N-terminal kinase/stress-activated protein kinase (JNK/SAPK) pathway, and the p38 pathway. From dendritic cell cDNA library, we isolated a full-length cDNA encoding a potentially novel 898-residue kinase, which was designated DPK. The protein contained a potential kinase domain at the N-terminal exhibiting homology with MEKK1-, MEKK2-, MEKK3-, MEKK4-, MEKK5-, Tpl-2-, and p21-activated kinases (PAKs), but no GTPase-binding domain which is characteristic of PAKs. Northern blotting analysis showed that DPK was ubiquitously expressed in normal tissues, with abundant expression in kidney, skeletal muscle, heart, and liver. When overexpressed in transfected NIH3T3 cells, it could activate both the ERK1/ERK2 pathway and the SAPK pathway in a dose-dependent manner, but not affect the p38 pathway. These findings suggested that DPK might be a novel candidate MAPKKK.

Amino Acid Sequence↗
PubMed2000

Anti-DNA antibodies exhibit different binding motif preferences for single stranded or double stranded DNA.

A common feature for most anti-DNA antibodies (Abs) is their induction in an antigen (Ag)-driven specific clonal expansion pattern though crossreactivity. However, the fine sequences in DNA Ags that interact directly with immune system and the ability of DNA to induce immune responses is poorly understood. In order to define the characteristics of possible antigenic determinants in DNA Ags, we immunized mice with the pBR322 plasmid and used antisera as source of anti-DNA Abs. A systemic evolution of ligands by exponential enrichment (SELEX) procedure was performed on an oligodeoxynucleotide library either in single stranded (ss-) or double stranded (ds-) form. The SELEXed fragments were cloned and sequenced. The resulting sequences were analyzed using the Multiple Alignment Construction and Analysis Workbench program. We show that the fragments of ss- or ds- form bound by a same stock of antibodies were different in their conserved sequences. ss-DNA fragments recognized by anti-DNA Abs were rich in cacc, caccc, accc or cccc blocks, while the same stock of Abs exhibited significant preference for the (5'gcg3'/3'cgc5') motif located in ds-DNA. At the same time sera from unimmunized control mice showed no sequence preference in either ss-DNA or ds-DNA. Future improvement of this work and the potential use of SELEX for studies of DNA Ags are also discussed.

Animals↗
PubMed2000

Characterization of the mouse and human PRSS17 genes, their relationship to other serine proteases, and the expression of PRSS17 in developing mouse incisors.

The human PRSS17 (serine protease 17) gene, which is located on chromosome 19q in a cluster of genes encoding serine proteases, has been variously designated enamel matrix serine proteinase 1 (EMSP1), prostase, KLK4, and KLK-L1. We have cloned and characterized the mouse and human PRSS17 genes. Both have six exons and five introns. The mouse PRSS17 gene sequence is 10134bp; the human sequence is 7115bp. Computer analysis of the mouse PRSS17 gene sequence upstream of the translation initiation codon identified two potential transcription initiation sites, at nucleotides 2878 and 2336. The first nucleotide of the reported mouse PRSS17 cDNA sequence corresponds to position 2352 on the gene, only 16 bases downstream from one of the putative transcription initiation sites. Repetitive DNA sequences from the MSR1 family are found in both the mouse and human PRSS17 genes. Additionally, the human PRSS17 gene contains Tigger2, MER8, and Alu repetitive sequences. Phylogenetic analyses of human and rodent proteases suggest that the PRSS17 protein is not a member of the kallikrein family of serine proteases but that the PRSS17 gene may have originated prior to the divergence of the kallikrein and trypsin families of proteases. To better characterize the timing of PRSS17 expression in developing teeth, we performed in-situ hybridization on postnatal day 3 developing mouse mandibular incisors. PRSS17 mRNA was not detected in secretory stage ameloblasts but could be detected in odontoblasts, while transition-stage and maturation-stage ameloblasts were strongly positive. This pattern supports a role for the PRSS17 protein in the degradation of enamel proteins.

Amino Acid Sequence↗
PubMed2000

Assessment of mitochondrial energy coupling in vivo by 13C/31P NMR.

The recently cloned uncoupling protein homolog UCP3 is expressed primarily in muscle and therefore may play a significant role in the regulation of energy expenditure and body weight. However, investigation into the regulation of uncoupling protein has been hampered by the inability to assess its activity in vivo. In this report, we demonstrate the use of a noninvasive NMR technique to assess mitochondrial energy uncoupling in skeletal muscle of awake rats by combining (13)C NMR to measure rates of mitochondrial substrate oxidation with (31)P NMR to assess unidirectional ATP synthesis flux. These combined (31)P/(13)C NMR measurements were performed in control, 10-day triiodo-l-thyronine (T(3))-treated (model of increased UCP3 expression), and acute 2,4-dinitrophenol (DNP)-treated (protonophore and mitochondrial uncoupler) rats. UCP3 mRNA and protein levels increased 8.1-fold (+/- 1.1) and 2.8-fold (+/- 0.8), respectively, in the T(3)-treated vs. control rat gastrocnemius muscle. (13)C NMR measurements of tricarboxylic acid cycle flux as an index of mitochondrial substrate oxidation were 61 +/- 21, 148 +/- 25, and 310 +/- 48 nmol/g per min in the control, T(3), and DNP groups, respectively. (31)P NMR saturation transfer measurements of unidirectional ATP synthesis flux were 83 +/- 14, 84 +/- 14, and 73 +/- 7 nmol/g per s in the control, T(3), and DNP groups, respectively. Together, these flux measurements, when normalized to the control group, suggest that acute administration of DNP (mitochondrial uncoupler) and chronic administration of T(3) decrease energy coupling by approximately 80% and approximately 60%, respectively, and that the latter treatment correlates with an increase in UCP3 mRNA and protein expression. This NMR approach could prove useful for exploring the regulation of uncoupling protein activity in vivo and elucidating its role in energy metabolism and obesity.

Adenosine Triphosphate↗
PubMed2000

Dynamic expression of glial cell line-derived neurotrophic factor after cerebral ischemia.

The aim of this study was to understand the possible involvement of glial cell line-derived neurotrophic factor (GDNF) in rat brain ischemic injury by examining the expression and the cellular location of GDNF with molecular biological and morphological techniques. Expression of GDNF mRNA and protein was first increased as early as 2h after ischemia-reperfusion in peri-infarct cerebral cortex and striatum; it then declined, and showed a second increase at 72 h. Double staining confirmed that the earlier peak of GDNF expression was of neuronal origin and the later peak of glial origin. Considering the neurotrophic characteristics of GDNF, our findings suggest that elevated endogenous GDNF expression may have important roles in protection of ischemic injured neuronal cells.

Animals↗
PubMed2000

Conserved plant genes with similarity to mammalian de novo DNA methyltransferases.

DNA methylation plays a critical role in controlling states of gene activity in most eukaryotic organisms, and it is essential for proper growth and development. Patterns of methylation are established by de novo methyltransferases and maintained by maintenance methyltransferase activities. The Dnmt3 family of de novo DNA methyltransferases has recently been characterized in animals. Here we describe DNA methyltransferase genes from both Arabidopsis and maize that show a high level of sequence similarity to Dnmt3, suggesting that they encode plant de novo methyltransferases. Relative to all known eukaryotic methyltransferases, these plant proteins contain a novel arrangement of the motifs required for DNA methyltransferase catalytic activity. The N termini of these methyltransferases contain a series of ubiquitin-associated (UBA) domains. UBA domains are found in several ubiquitin pathway proteins and in DNA repair enzymes such as Rad23, and they may be involved in ubiquitin binding. The presence of UBA domains provides a possible link between DNA methylation and ubiquitin/proteasome pathways.

Amino Acid Sequence↗
PubMed2000

Asymmetric requirements for a Rab GTPase and SNARE proteins in fusion of COPII vesicles with acceptor membranes.

Soluble NSF attachment protein receptor (SNARE) proteins are essential for membrane fusion in transport between the yeast ER and Golgi compartments. Subcellular fractionation experiments demonstrate that the ER/Golgi SNAREs Bos1p, Sec22p, Bet1p, Sed5p, and the Rab protein, Ypt1p, are distributed similarly but localize primarily with Golgi membranes. All of these SNARE proteins are efficiently packaged into COPII vesicles and suggest a dynamic cycling of SNARE machinery between ER and Golgi compartments. Ypt1p is not efficiently packaged into vesicles under these conditions. To determine in which membranes protein function is required, temperature-sensitive alleles of BOS1, BET1, SED5, SLY1, and YPT1 that prevent ER/Golgi transport in vitro at restrictive temperatures were used to selectively inactivate these gene products on vesicles or on Golgi membranes. Vesicles bearing mutations in Bet1p or Bos1p inhibit fusion with wild-type acceptor membranes, but acceptor membranes containing these mutations are fully functional. In contrast, vesicles bearing mutations in Sed5p, Sly1p, or Ypt1p are functional, whereas acceptor membranes containing these mutations block fusion. Thus, this set of SNARE proteins is symmetrically distributed between vesicle and acceptor compartments, but they function asymmetrically such that Bet1p and Bos1p are required on vesicles and Sed5p activity is required on acceptor membranes. We propose the asymmetry in SNARE protein function is maintained by an asymmetric distribution and requirement for the Ypt1p GTPase in this fusion event. When a transmembrane-anchored form of Ypt1p is used to restrict this GTPase to the acceptor compartment, vesicles depleted of Ypt1p remain competent for fusion.

Alleles↗
PubMed2000

Smad6 as a transcriptional corepressor.

Smad6 and Smad7, a subgroup of Smad proteins, antagonize the signals elicited by transforming growth factor-beta. These two Smads, induced by transforming growth factor-beta or bone morphogenetic protein (BMP) stimulation, form stable associations with their activated type I receptors, blocking phosphorylation of receptor-regulated Smads in the cytoplasm. Here we show that Smad6 interacts with homeobox (Hox) c-8 as a transcriptional corepressor, inhibiting BMP signaling in the nucleus. The interaction between Smad6 and Hoxc-8 was identified by a yeast two-hybrid approach and further demonstrated by co-immunoprecipitation assays in cells. Gel shift assays show that Smad6, but not Smad7, interacts with both Hoxc-8 and Hoxa-9 as a heterodimer when binding to DNA. More importantly, the Smad6-Hoxc-8 complex inhibits interaction of Smad1 with Hoxc-8- and Smad1-induced transcription activity. These data indicate that Smad6 interacts with Hox transcription factors as part of the negative feedback circuit in the BMP signaling pathway.

Bone Morphogenetic Proteins↗
PubMed2000

DNA immunization utilizing a herpes simplex virus type 2 myogenic DNA vaccine protects mice from mortality and prevents genital herpes.

A gene transfer vector for DNA immunization was developed in which the promoter was derived from the murine muscle creatine kinase (MCK) gene; a gene expressed only in differentiated skeletal muscle. In vitro, we observed high-level, but unrestricted, gene expression from the cytomegalovirus (CMV) promoter unlike expression from the MCK promoter which was weak but restricted to myofibers. A myogenic DNA vaccine (MDV) that encoded the glycoprotein D gene from herpes simplex virus type-2 (HSV-2) was used to DNA immunize mice. MDV immunization resulted in virus specific immunity that protected HSV-2 infected mice from mortality and prevented the development of genital herpes. Therefore, we conclude that high-level gene expression or the use of a strong transcription unit was not a prerequisite for an efficacious DNA vaccine and the use of a nonviral tissue specific promoter could suffice.

Animals↗