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Biomedical subjects

C Yan

Publications and source records attributed to C Yan.

At least 127 records · Page 7Linked to original sources

Upstream enhancer activity in the human surfactant protein B gene is mediated by thyroid transcription factor 1.

Surfactant protein B (SP-B) is selectively expressed in bronchiolar and alveolar epithelial cells of the lung. We identified an upstream enhancer located in the 5'-flanking region of the human SP-B gene (-439 to -331 base pair, hSP-B(-439/-331)) by deletion analysis of SP-B-luciferase constructs assessed in transfection assays in vitro. The element cis-activated the expression of an SV40 promoter-luciferase reporter gene in a human pulmonary adenocarcinoma cell line (H441-4). Three distinct binding sites for the nuclear transcription protein, thyroid transcription factor 1 (TTF-1), were identified, and the purified TTF-1 homeodomain was bound to bhe region of hSP-B(-439/-331). Co-transfection of H441-4 cells with the expression vector pCMV-TTF-1 trans-activated the native human SP-B promoter and the SV40 promoter fused with the SP-B enhancer. Mutations of the TTF-1 binding sites in the upstream enhancer blocked TTF-1 binding and transactivation activity. In summary, TTF-1 interacts with distinct proximal (-80 to -110) and distal (-439 to -331) cis-acting elements than regulate lung epithelial cell-specific transcription of the human SP-B gene.

Base Sequence↗

Molecular cloning and characterization of a calmodulin-dependent phosphodiesterase enriched in olfactory sensory neurons.

The sensing of an odorant by an animal must be a rapid but transient process, requiring an instant response and also a speedy termination of the signal. Previous biochemical and electrophysiological studies suggest that one or more phosphodiesterases (PDEs) may play an essential role in the rapid termination of the odorant-induced cAMP signal. Here we report the molecular cloning, expression, and characterization of a cDNA from rat olfactory epithelium that encodes a member of the calmodulin-dependent PDE family designated as PDE1C. This enzyme shows high affinity for cAMP and cGMP, having a Km for cAMP much lower than that of any other neuronal Ca2+/calmodulin-dependent PDE. The mRNA encoding this enzyme is highly enriched in olfactory epithelium and is not detected in six other tissues tested. However, RNase protection analyses indicate that other alternative splice variants related to this enzyme are expressed in several other tissues. Within the olfactory epithelium, this enzyme appears to be expressed exclusively in the sensory neurons. The high affinity for cAMP of this Ca2+/calmodulin-dependent PDE and the fact that its mRNA is highly concentrated in olfactory sensory neurons suggest an important role for it in a Ca(2+)-regulated olfactory signal termination.

3',5'-Cyclic-AMP Phosphodiesterases↗

Developmentally expressed Ca(2+)-sensitive adenylyl cyclase activity is disrupted in the brains of type I adenylyl cyclase mutant mice.

The type I Ca(2+)-sensitive adenylyl cyclase has been implicated in several forms of synaptic plasticity in vertebrates. Mutant mice in which this enzyme was inactivated by targeted mutagenesis show deficient spatial memory and altered long term potentiation (Wu, Z. L., Thomas, S. A., Villacres, E. C., Xia, Z., Simmons, M. L., Chavkin, C., Palmiter, R. D., and Storm, D. R. (1995) Proc. Natl Acad Sci. U. S. A. 92, 220-224). Long term potentiation in the CA1 region of the rat hippocampus develops during the first 2 weeks after birth and reaches maximal expression at postnatal day 15 with a gradual decline at later stages of development. Here we report that Ca(2+)-stimulated adenylyl cyclase activity in rat hippocampus, cerebellum, and cortex increases significantly between postnatal days 1-16. This increase appears to be due to enhanced expression of type I adenylyl cyclase rather than type VIII adenylyl cyclase, the other adenylyl cyclase that is directly stimulated by Ca2+ and calmodulin. Type I adenylyl cyclase mRNA in the hippocampus increased 7-fold during this developmental period. The developmental expression of Ca(2+)-stimulated adenylyl cyclase activity in mouse brain was attenuated in mutant mice lacking type I adenylyl cyclase. Changes in expression of the type I adenylyl cyclase during the period of long term potentiation development are consistent with the hypothesis that this enzyme is important for neuroplasticity and spatial memory in vertebrates.

Adenylyl Cyclases↗

Dependence of ricin toxoid vaccine efficacy on the structure of poly(lactide-co-glycolide) microparticle carriers.

Biodegradable microparticles made of poly(lactide-co-glycolide) (PLG) were used for protracted and pulsed-release of the incorporated ricin toxoid (RT) vaccine to reduce the multiple immunization doses and the time required to induce complete protection against lethal aerosol-borne ricin challenge. The release rate of RT encapsulated in PLG microparticles was controlled by polymer selection and varying the preparation procedures, which allowed us to control microparticle size and the distribution of the vaccine in the polymeric matrix. PLG-microparticles in which RT vaccine was distributed heterogeneously in small pockets stimulated a rapid antibody response which was independent of the polymeric composition of the carriers. PLG-microparticles in which RT vaccine was distributed homogeneously throughout the polymeric matrix induced a slower antibody response, which depended on the polymeric composition of the carriers. Administration of RT in homogeneous microparticles made from 50/50 PLG or 100% polylactide stimulated two distinct anti-ricin IgG peaks, while RT in heterogeneous microparticles stimulated identical IgG peaks. An early (3 weeks) and long-lasting (1 year or longer) anti-ricin antibody response was evoked by a single administration of encapsulated RT vaccine when prepared by the above-mentioned conditions. In contrast, three administrations of the aqueous RT were required to stimulate similar antibody response. Reduction of immunization time from 6 to 4 weeks was achieved with RT encapsulated in small homogeneous microparticles but not with homogeneous large microparticles. These results demonstrated the usefulness of biodegradable microparticles to improve the efficacy of immunization with RT vaccine and probably many other vaccines as well.

Animals↗

Effects of local hypothermia on uveal blood flow and postoperative inflammation in vitrectomy.

Effects of local hypothermia on uveal blood flow and postoperative inflammation were evaluated in experimental vitrectomy in albino rabbits. Solutions used for intraocular perfusion were maintained at 9 degrees C, 22 degrees C or 37 degrees C. Following closed vitrectomy, the vitreous cavity was irrigated for 60 minutes. Temperatures at various sites and uveal blood flow were measured before and during the procedure. Aqueous protein concentrations were checked on postoperative days 1, 7 and 14. There was a larger decrease in temperature at the retina than at the choroid or the ciliary body. Blood flow at the ciliary body was reduced to 76.0% and that at the choroid to 77.0% of the preoperative level after 60 minutes of irrigation at 9 degrees C. The decrease was 91.0% and 88.3%, respectively, after 60 minutes of irrigation at 22 degrees C. Aqueous protein concentrations in the 9 degrees C and 22 degrees C groups were significantly lower than those in the 37 degrees C group on the first postoperative day in the eyes irrigated for 60 minutes. In the eyes irrigated for 30 minutes, however, no significant differences were seen. Local hypothermia during prolonged vitrectomy seems to decrease inflammation in the early postoperative stage.

Animals↗

Detection of congenital cytomegalovirus infection by using chorionic villi of the early pregnancy and polymerase chain reaction.

OBJECTIVE: To detect congenital cytomegalovirus (CMV) infection of chorionic villi in early pregnancy. METHODS: Extraction of DNA of chorionic villi and amplification of the gene of major immediate-early (MIE) antigen of CMV using a polymerase chain reaction (PCR). RESULTS: Sixty-eight specimens of chorionic villi and 16 specimens were positive for CMV infection by PCR. The incidence of congenital CMV infection in the first trimester of pregnancy was 23.5%. CONCLUSIONS: The risk of transmission of CMV from mother to fetus in early pregnancy is very high and potential CMV carriers may transmit CMV to their fetus in early pregnancy.

Chorionic Villi↗

Peptide V: a VGF-derived neuropeptide purified from bovine posterior pituitary.

The objective of this study was to purify PRL-releasing factor (PRF) from the bovine posterior pituitary (PP) and determine its structure. Five hundred bovine PPs were acid extracted and fractionated using gel filtration chromatography followed by semipreparative and analytical HPLC. PRF activity was determined by an in vitro bioassay. After six chromatographic steps, a single peak with PRF activity was resolved. As determined by mass spectrometry and microsequencing, this peak contained a major peptide composed of 30 amino acids with a mol wt of 3708K. A synthetic peptide was then produced by solid-phase synthesis. When tested both in vivo and in vitro, the synthetic peptide lacked PRF activity. Further HPLC fractionation under different conditions resolved the synthetic peptide from a highly purified PRF activity. This indicated that the isolated peptide was coincidentally eluted with PRF during the purification. The major isolated peptide has 94% identity with a sequence at the C-terminus of a rat protein named VGF. VGF is a nerve growth factor-inducible protein that has been identified in PC12 cells and is localized in selected sites throughout the central nervous system. The isolated peptide has an Arg-Arg cleavage site at its junction within the VGF protein. Based on this information, we named this substance Peptide V (VGF-derived peptide). We postulate that Peptide V is: 1) a natural cleavage product of the VGF protein; 2) produced and processed either in the hypothalamus or within the pituitary proper, and 3) a releasable peptide that fulfills one or more endocrine functions.

Animals↗

[Hypothermic effects on uveal blood flow and postoperative inflammation in vitrectomy].

Hypothermic effects on uveal blood flow and postoperative inflammation were evaluated in experimental vitrectomy on albino rabbits. Solutions used for intraocular perfusion were maintained at 9 degrees C and 22 degrees C. Following the vitrectomy, the rabbits' intraocular spaces were irrigated for 60 minutes. The temperature at various sites and the uveal blood flow were measured before and during the procedure. Aqueous protein concentrations were checked on postoperative days 1, 7, and 14. Another solution maintained at 37 degrees C was used as a control. There was a larger decrease in temperature at the retina than there was at the choroid and the ciliary body. Blood flow at the ciliary body decreased to 76.0% and 77.0% at the choroid after 60 minutes of irrigation at 9 degrees C. The decrease at 22 degrees C were 91.0% and 88.3%, respectively. Aqueous protein concentrations at both 9 degrees C and 22 degrees C were significantly lower than at 37 degrees C on the first postoperative day in the eyes irrigated for 60 minutes. In the eyes irrigated for 30 minutes, however, no significant differences were seen. Hypothermia during a prolonged vitrectomy operation seems to decrease inflammation in early postoperative stages.

Animals↗

[Significance of surgical treatment for bone metastasis from lung cancer].

To assess significance of surgical therapy for patients who had metastatic bone tumor from primary lung cancer, sites of recurrence in patients who underwent surgical resection for lung cancer lesions were analyzed. Clinical courses of patients who underwent surgical therapy for metastatic bone tumor were analyzed, too. 1) From 1985 to 1989, 88 patients died from lung cancer in our hospital who has underwent pulmonary resection for lung cancer lesions. Of these, 22 patients had metastatic bone tumor. Sixty patients died who had underwent complete resection for lung cancer lesions. In eight patients of these 60 cases, bone were first site of recurrence. Of these eight patients, length from pulmonary resection to detection of bone metastasis were 4-23 months. In the other hand, length from detection of bone metastasis of death were 3-24 months. There was no correlation between two length. All of these eight patients had another metastatic lesions at the time of death. 2) From 1985 to July 1993, twelve patients underwent surgical therapy for their bone metastasis of primary lung cancer. Of these, four had metastatic cancer in thoracic vertebral body, two had in clavicle, two had in rib, three had in femoral bone, another had in tibial bone. Objective of operation were improvement of paralysis or pain, prevention of fracture, and so on. Most cases succeeded to get improvement in a short time, but some cases got worse soon. Surgical treatment was effective to improve paralysis or pain, and to increase quality of life. But it can not be effective except appropriate management in a short time because another symptom raise soon.

Aged↗

Differential expression of the 61 kDa and 63 kDa calmodulin-dependent phosphodiesterases in the mouse brain.

Based on their relative abundance and regulation by Ca2+ and by phosphorylation in vitro, it is thought that the Ca2+/calmodulin-dependent phosphodiesterases (CaM-PDEs) are important modulators of cyclic nucleotide function in the brain. Two of the most abundant CaM-PDEs in the brain are the 61 kDa and 63 kDa isozymes. In this study, the regional and cellular expression of mRNA encoding these two different isoforms in mouse brain has been determined by in situ hybridization. The 63 kDa CaM-PDE mRNA has a wide-spread but uneven distribution. Very strong hybridization signals are present in the caudate-putamen, nucleus accumbens, olfactory tubercle, and dentate gyrus of the hippocampus. Somewhat lesser amounts of 63 kDa CaM-PDE mRNA are present in the olfactory bulb and piriform cortex. Weaker but still easily discernible hybridization signals are seen in several layers of the cerebral cortex, CA1 and CA3 regions of the hippocampus, amygdaloid nuclear complex, thalamus, hypothalamus, midbrain, brainstem, cerebellum, and spinal cord. A weak hybridization signal was detected in the globus pallidus of the basal ganglia. In general, the distribution of the 63 kDa CaM-PDE is very similar to that of dopamine receptors, suggesting that it may modulate dopamine function. In contrast, the 61 kDa CaM-PDE mRNA has a more limited and much different distribution, with the highest level of expression in the cerebral cortex and in the pyramidal cells of the hippocampus. A moderate hybridization signal was detected in the medial habenula and amygdaloid nuclear complex. In addition, small subsets of neurons in several other regions showed specific hybridization. Both PDE mRNAs appear to be localized exclusively in neuronal cell bodies. Their distinct distribution suggests important but different physiological roles for these two isozymes in the regional regulation of cyclic nucleotides in the CNS. Since these two isozymes are differentially phosphorylated by cAMP-dependent and Ca2+/CaM-dependent protein kinases, the differential expression also provides a potential mechanism by which these PDEs can differentially regulate cAMP and cGMP in different brain areas. The high expression levels in specific subsets of neurons also suggest that agents increasing Ca2+ in these neurons will increase the rate of cyclic nucleotide degradation.

Animals↗

Multiple regions of NSR1 are sufficient for accumulation of a fusion protein within the nucleolus.

NSR1, a 67-kD nucleolar protein, was originally identified in our laboratory as a nuclear localization signal binding protein, and has subsequently been found to be involved in ribosome biogenesis. NSR1 has three regions: an acidic/serine-rich NH2 terminus, two RNA recognition motifs, and a glycine/arginine-rich COOH terminus. In this study we show that NSR1 itself has a bipartite nuclear localization sequence. Deletion of either basic amino acid stretch results in the mislocation of NSR1 to the cytoplasm. We further demonstrate that either of two regions, the NH2 terminus or both RNA recognition motifs, are sufficient to localize a bacterial protein, beta-galactosidase, to the nucleolus. Intensive deletion analysis has further defined a specific acidic/serine-rich region within the NH2 terminus as necessary for nucleolar accumulation rather than nucleolar targeting. In addition, deletion of either RNA recognition motif or point mutations in one of the RNP consensus octamers results in the mislocalization of a fusion protein within the nucleus. Although the glycine/arginine-rich region in the COOH terminus is not sufficient to bring beta-galactosidase to the nucleolus, our studies show that this domain is necessary for nucleolar accumulation when an RNP consensus octamer in one of the RNA recognition motifs is mutated. Our findings are consistent with the notion that nucleolar localization is a result of the binding interactions of various domains of NSR1 within the nucleolus rather than the presence of a specific nucleolar targeting signal.

Amino Acid Sequence↗

A domestic cell bioreactor and its application in virus culture.

A cell culture bioreactor (CellCul-20) and its application in cell and virus culture are described in this paper. It has been evaluated with strict aseptic tests and one-year's operation shown that CellCul-20 bioreactor can keep its aseptic condition after being autoclaved. It can meet the requirement for the control of the main parameters for cell and virus culture and the finely adjustment of the main parameters to meet the changing conditions of the cultivation. A high cell density and a high level of virus titre were reached respectively for Vero cells and Japanese encephalitis virus (JEV) while they were cultured in this bioreactor. It is the first report on large-scale culture of JEV-infected Vero cells to prepare primary JEV vaccine. Some suggestions are made for the improvement of CellCul-20.

Animals↗

Molecular cloning and characterization of additional factors that bind to the interferon-alpha/beta response element B of the murine (2'-5')oligoadenylate synthetase ME-12 gene.

The function of interferon (IFN) response elements (IREs) may be classified into two classes according to the location of the IRE in the polymerase II transcription promoter: located upstream of the core promoter, or located within the core promoter. Thus, the IRE-binding factors (IREBFs) can serve as sequence-specific transcription factors that activate IFN-inducible gene transcription and they can participate in preinitiation complex formation. In the murine and human (2'-5')oligoadenylate synthetase genes the IRE is located within the core promoter region. For this reason, it is important to isolate cDNAs encoding protein factors that can bind to this IRE and determine their functional roles in the regulation of expression of the IFN-inducible (2'-5')oligoadenylate synthetase gene. We have isolated three groups of cDNAs that encode IREBFs. Here we report on clone 38 cDNA, which contains an open reading frame encoding a protein of 277 amino acids that is designated IREBF-2. The C terminus of IREBF-2 is extremely proline-rich and contains a high percentage of short-armed amino acids. IREBF-2 shares marked similarity with some gene products of the herpes group of viruses. These gene products are potent transcription activators. IREBF-2 is constitutively expressed in BALB/c 3T3 cells.

2',5'-Oligoadenylate Synthetase↗

Identification of a new interferon-alpha/beta-inducible DNA-binding protein that interacts with the regulatory element A of 2',5'-oligoadenylate synthetase ME-12 gene.

A new interferon (IFN)-stimulated response factor (ISRF) has been identified in nuclear extracts of IFN-alpha/beta-treated murine BALB/c-3T3 fibroblasts by the mobility-shift electrophoresis assay. The factor, ISRF-2, displays murine 2',5'-oligoadenylate (2-5A) synthetase ME-12 gene 5' regulatory element A specificity and differs from the previously described IFN response element B-specific factor ISRF-1 in several aspects. ISRF-2 is restricted to the nucleus, whereas ISRF-1 exists in the cytoplasm and translocates into the nucleus upon treatment of cells with IFN-alpha/beta. The ionic strength requirement of ISRF-2 for maximal DNA-binding activity is lower than that of ISRF-1. The DNA-binding activity of ISRF-2, but not that of ISRF-1, is markedly suppressed by Mg2+. In common with ISRF-1, the phosphorylated form of ISRF-2 appears to be required for DNA-binding activity. A model is proposed for the mechanism whereby murine IFN-alpha/beta regulates 2-5A synthetase ME-12 gene expression.

2',5'-Oligoadenylate Synthetase↗

Evaluation of the effect of PTCR/PTCA by 99mTc-MIBI myocardial imaging in acute and old myocardial infarction.

This study is to clarify the ability of technetium-99m-Hexakis-2-methoxyisobutyl isonitrile(MIBI) myocardial scintigraphy for assessment of the effect of PTCR/PTCA as compared with conventional thallium-201 myocardial scintigraphy (201Tl). 99mTc-MIBI (740-888MBq) was injected before emergency PTCR/PTCA, and the initial imaging was performed immediately after PTCR/PTCA in 4 patients with acute myocardial infarction (AMI). On the other hand, in 2 patients with old myocardial infarction (OMI), 201Tl and MIBI myocardial imagings were performed before and one week after rescue PTCA. The defect area on the follow-up MIBI imaging was smaller than that on the initial MIBI imaging in 2 patients with successful emergency PTCR/PTCA. But in the other 2 AMI patients with unsuccessful reperfusion, the defect area did not significantly decrease after PTCR/PTCA. Reperfusion was successful in one of the two patients with OMI. In both the two patients with OMI, the MIBI defect area did not significantly decrease after rescue PTCA. MIBI myocardial scintigraphy is a useful noninvasive method for evaluating the effect of emergency PTCR/PTCA for myocardial salvage in patients with AMI, because MIBI is a kit type agent, and it does not redistribute.

Aged↗

[Hepatic carcinoma treated by hepatic arterial embolization using 131I and chemotherapeutic agent gelatin microspheres: report of 9 cases].

Nine patients with inoperable hepatoma were treated by using hepatic arterial embolization 131I and chemotherapeutic agent gelatin microsphere (131I-CA-GM). The emission CT after operation detected that the microspheres were concentrated on tumor area. The ratio between the radioactivity in tumor and that in liver was 4.1:1. A case died of ictopic embolization; the others survived 3, 4, 5, 19, 24, 7, 8, and 12 months respectively. Three of them were still alive. 131I-CA-GM has triple anticarcinogenic actions, including the arterial occlusion, targeting chemotherapy and internal radiation. The microspheres can selectively accumulate in the tumor artery and can be easily traced by gamma-camera or emission CT. 131I-CA-GM is a hopeful embolic agent for the treatment of liver cancer, but some problems about ectopic arterial embolization should be further studied.

Carcinoma, Hepatocellular↗

A method for studies of an El Tor-associated antigen of Vibrio cholerae O1.

A method for studying the biotype El Tor associated mannose-sensitive haemagglutinin (MSHA) of V. cholerae O1 has been developed. By using crude MSHA adsorbed to chicken erythrocytes as solid phase antigen in an enzyme-linked immunosorbent assay (ELISA), antisera against V. cholerae of the El Tor biotype reacted in high titre with the MSHA-coated cells, whereas antisera against vibrios of the classical biotype did not bind significantly, i.e. in higher titre than pre-immune sera. The binding of anti-MSHA serum, or a monoclonal antibody against MSHA, to the MSHA-coated erythrocytes could be efficiently inhibited by crude MSHA as well as by El Tor vibrios whereas neither V. cholerae lipopolysaccharide nor different strains of classical vibrios had any inhibitory effect. These results support the existence of an El Tor-associated immunogen. They also suggest a possibility of determining antibodies against different haemagglutinins in ELISA without having access to purified antigens.

Animals↗