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Biomedical subjects

W Yang

Publications and source records attributed to W Yang.

At least 433 records · Page 24Linked to original sources
PubMed1996

[Research on the recombinant plasmid pDJH2 of L. interrogans serovar lai: sequencing and alignment with other known bacterial Omp sequence].

The Leptospira whole cell vaccine (LWCV) currently used in China is safe and effective, out the immunity following vaccination with two doses of the fluid medium vaccine is of low order. The duration of immunity conferred by this vaccine is rather short, six months or at most one year. Therefore, it is necessary to develop new generation vaccines against Leptospirosis for the developing world. In this paper we report the sequencing of the insert fragment of pDJH2 from genomic DNA of L. interrogans sevovar lai strain 017 and its alignment with other bacterial omp sequences. A genomic library of Leptospira interrogaans serovar lai strain 017 was constructed with the plasmid vector pUC18. A recombinant plasmid designated pJDH2 was screened from the genomic library. Inserted fragment of pDH2 is 1.9 kb by gel electrophoresis. Immunization/protection was studied in BALB/c mice model. The results showed highly significant difference between pDJH2 and pUC18 (control). Inserted fragment of pDJH2 DNA sequencing was performed by Dr Yan Zhengxin (Max-Planck-Institut for Biology. Tubingen, Germany). Insert fragment was cloned into pBluescript II KS-(stratagene) and sequenced by using AB1 (Applied Bio Systems, Model 373A). Two open reading frames of 565 and 662 nucleotides were identified. There were identifiable initiation codons, terminators, Shine-Dalgano ribosome combining site, Pribnow boxes and Sextama boxes within the 2 sequenced regions. Nucleotide sequences were analysed using Gene Work, a suit of computer program developed by Department of Biochemistry St. Jude Children's Research Hospital Memphis. U.S.A. The results of formatted alignment showed the predicted nucleotide sequence of ORF1 of the serovar lai had significant similarity with ORF2 (49.36%). L. kirschneri ompL1 (49.26%), Borrelia burgdoferi omp (48.97%), Treponema phagedenis omp (47.3%); Salmonella typhimurium ompC(46.87%), Yersinia enterocolitica ompH (46.7%), Leptospira borgpeterseni pfap (46.3%), and Serratia marcescens omp (43.3%). The close relationship of the pDJH2 ORF1 and ORF2 nucleotide sequences from Leptospira kirschneri ompL 1 is apparent. Whether the recombinant pDJH2 will prove useful for vaccine development remains to be tested.

Animals↗
PubMed1996

[The study on noninvasive evaluation of pulmonary artery pressure by the method of pulmonary artery flow spectrum in chronic pulmonary disease].

OBJECTIVE: Pulmonary artery flow spectrum of patients with chronic pulmonary disease were studied in order to find excellent Doppler Indexes reflected pulmonary artery pressure, and establish multiple regression equation for estimating pulmonary artery pressure more accurately. METHODS: During catheterization, 54 patients with chronic pulmonary disease were examined by echocardiography. Fourteen Doppler indexes of spectrum were compared with the pulmonary artery pressure by catheter simultaneously, and multiple regression equations were taken. RESULTS: There were significant correlation between Doppler indexes PEP/AcT, AcVm, VI/(QTxVmax), RVET/QT and pulmonary artery pressure. Two multiple regression equations were deduced: PAPs (kPa) = [9.4 x PEP/AcT-1.4 x AcVm-126.1 x VI/(QT x Vmax) + 75] divided by 7.5 and PAPm (kPa) = [7.5 x PEP/AcT-1.2 x AcVm- 87.7 x VI/(QT X Vmax) + 48] divided by 7.5. CONCLUSIONS: Above two regression equations could be used to noninvasively evaluate pulmonary artery pressure in patients with chronic pulmonary disease.

Adult↗
PubMed1996

[Failure in mastoidectomy with posterior canal wall preservation and tympanoplasty case analysis].

Preservation of the posterior canal wall in mastoidectomy is essential for rebuilding normal tympanum and conductive structures, and has been widely recommended. However, controversies remain about the technique due to its complications and diverse results reported. This paper analysed the causes of four failed cases in detail and proposed measure to reduce morbidities and promote success. The review includes case selection, operation staging, surgical techniques and lesion removal etc.

Adult↗
PubMed1996

[Histopathology of the temporal bone in adhesive otitis media].

To understand the histopathological changes of adhesive otitis media (AOM) in human, temporal bone sections processed with the celloidin-embedding technique were reviewed. In the 338 pairs of temporal bone specimens, 20 cases (26 ears) showed characteristics of AOM. The results showed that the lesions of AOM were extensive, involving all middle ear structures including tympanic muscles, round and oval windows. There were various pathological changes, in every stage of inflammation and tissue repair. According to the main pathological characteristics, histopathological changes of the 26 ears were staged as exudation, inflammatory grannlation formation, histocyte response and collegen proliferation. As the process progressed, the amount of inflammatory cells and exudate decreased with the increase of fibroblasts. Certain factors were speculated to play an important role in the process of fibrosis. The authors analysed the difficulties in the management of AOM and causes of the unsatisfactory therapeutic results. Possible measures to improve treatment results were proposed.

Adult↗
PubMed1996

[CT study of the ostiomeatal complex in patients with deviated nasal septum].

This study was designed to evaluate the influence of nasal septal deviation on ostiomeatal complex (OMC). From April 1994 to February 1995, 32 patients with deviated nasal septum were investigated by high-resolution CT scan. The results showed that (1) 15 patients (15/32) were found to have ipsilateral or bilateral ostiomeatal complex disease (OMCD). The incidence of OMCD was related to the severity of the septal deviation; (2) measurement of uncinate process angulation and ethmoid infundibulum width in patients with OMCD was of limited value; (3) the relationship between septal deviation and OMC variation needed further study. It has been indicated that for patients with septal deviation coronal CT study is necessary to evaluate the degree of OMCD.

Adult↗
PubMed1996

[Analysis of cinnamic acid in storax and its original plant by HPLC].

The contents of free and hydrolytic cinnamic acid in storax and its original plant (Liquidambar orientalis) were determined by high performance liquid chromatography, and the chromatographic behaviors were studied. In styrax gummi, the contents of free and hydrolytic cinnamic acid were 7.03% and 25.26% respectively; and in its original plant, they were 0.3% evenly.

Chromatography, High Pressure Liquid↗
PubMed1995

Cloning and characterization of the human GABAA receptor alpha 4 subunit: identification of a unique diazepam-insensitive binding site.

Benzodiazepines modulate gamma-aminobutyric acid (GABA)-evoked chloride currents through a specific binding site at the GABAA receptor-chloride channel complex. The heterogeneity of diazepam-sensitive benzodiazepine binding sites (type I and type II) has been identified by pharmacological approaches both with native receptors and recombinant receptors coexpressing alpha, beta and gamma subunits. In addition, two distinguishable diazepam-insensitive benzodiazepine sites are found, spatially distributed between cerebral cortical and cerebellar regions. Coexpression of alpha 6 with beta 2 and gamma 2L subunits creates a pharmacologically similar benzodiazepine receptor to the diazepam-insensitive site observed in cerebellum, however, there is no evidence regarding the possible subunit combination forming the DI site in cerebral tissues. Here we report the cloning of the human alpha 4 cDNA and its pharmacology by coexpression of this alpha 4 subunit with beta 2 and gamma 2L subunits. This recombinant receptor complex showed a high affinity for the previously described benzodiazepine partial agonist bretazenill, the pyrazoloquinoline compounds CGS-9895 and CGS-9896, as well as the inverse agonists DMCM (methyl 6,7-dimethoxy 4-ethyl-beta-carboline-3-carboxylate) and Ro15-4513 as determined by [3H]Ro15-4513 binding. However, it is insensitive to the benzodiazepine type I selective compounds CL218.872 (3-methyl-6-[3-(trifluoromethyl)[phenyl]-1,2,4-triazolo[4.3-b]pyridazine ) and zolpidem as well as the benzodiazepine full agonists diazepam, halazolam and midazolam. In addition, the benzodiazepine receptor ligands DMCM, beta-CCE (beta-carboline-3-carboxylate ethyl ester), Beta-CCM (beta-carboline-3-carboxylate methyl ester), FG-7142, CGS-9895 and CGS-9896 showed 7 to 10 times higher affinity for alpha 4 beta 2 gamma 2L. The pharmacology of the alpha 4 beta 2 gamma 2L receptor complex appears to resemble those of the diazepam-insensitive site found in the cerebral cortex. Our study thus suggests that this subpopulation of diazepam-insensitive GABAA receptors may be composed of alpha 4 beta 2 gamma 2L subunits.

Amino Acid Sequence↗
PubMed1995

Susceptibility of idarubicin, daunorubicin, and their C-13 alcohol metabolites to transport-mediated multidrug resistance.

The intracellular pharmacokinetics and cytotoxicity of idarubicin (IDA), daunorubicin (DNR), and their corresponding C-13 alcohol metabolites, idarubicinol (IDAol) and daunorubicinol (DNRol), were studied in drug-sensitive HL-60/W human leukemia cells, and in two multidrug-resistant (MDR) sublines, HL-60/Vinc (overexpress P-glycoprotein, Pgp) and HL-60/Adr (overexpress multidrug resistance-associated protein, MRP). Intracellular drug accumulation (1 micrograms/mL) and retention were measured by flow cytometry. Mean intracellular steady-state concentration (Css, fluorescence units/cell) and area under the intracellular drug concentration x time curve (AUC, Fl.U/cell.min) were calculated. Relative to the values for the respective drugs in HL-60/W cells, the Css and AUC of IDA were much higher than those of DNR in the MDR cell lines, with Css and AUC of IDAol intermediate between IDA and DNR. In the MDR cell lines, the MDR modulator cyclosporine A (CsA), in concentrations of 0.3 to 30 mumol/L, caused minimal effects on 3-hr IDA accumulation, intermediate enhancement of IDAol accumulation, and greatest enhancement of DNR accumulation. The MDR cell lines were much less resistant to IDA (3- to 16-fold) than to DNR (65- to 117-fold). This difference was not the result of IDA being more potent than DNR, since the sensitivity of HL-60/W cells to IDA differed from their sensitivity to DNR by < 2-fold. The cellular pharmacokinetics and cytotoxicity of IDA in MDR human breast carcinoma cells MCF-7/AdrVp, which overexpress the putative MDR transporter P-95, were far superior to those of DNR, and were comparable to these parameters for IDA in parental MCF-7/W cells. These studies demonstrate that the cellular pharmacology and cytotoxicity of IDA in MDR cell lines that overexpress MRP, Pgp, or P-95 are more advantageous than those of DNR, suggesting that IDA is less susceptible to the transport-mediated MDR mechanism manifested. IDA is not completely invulnerable to MDR, however, since the MDR sublines studied did display a demonstrable level of resistance to IDA, compared with their drug-sensitive counterparts. IDAol, the major plasma metabolite of IDA, demonstrated behavior intermediate between the MDR-susceptible drug DNR and its parent compound, suggesting that its cytotoxic action is subject to transport-mediated cellular defenses.(ABSTRACT TRUNCATED AT 400 WORDS)

ATP Binding Cassette Transporter, Subfamily B, Mem↗
PubMed1995

The association of lipid activators with the amphipathic helical domain of CTP:phosphocholine cytidylyltransferase accelerates catalysis by increasing the affinity of the enzyme for CTP.

The biochemical mechanism for the regulation of enzyme activity by lipid modulators and the role of the amphipathic alpha-helical domain of CTP:phosphocholine cytidylyltransferase (CT) was investigated by analyzing the kinetic properties of the wild-type protein and two truncation mutants isolated from a baculovirus expression system. The CT[delta 312-367] mutant protein lacked the carboxyl-terminal phosphorylation domain and retained high catalytic activity along with both positive and negative regulation by lipid modulators. The CT[delta 257-367] deletion removed in addition the region containing three consecutive amphipathic alpha-helical repeats. The CT[delta 257-367] mutant protein exhibited a significantly lower specific activity compared to CT or CT[delta 312-367] when expressed in either insect or mammalian cells; however, CT[delta 257-367] activity was refractory to either stimulation or inhibition by lipid regulators. Lipid activators accelerated CT activity by decreasing the Km for CTP from 24.7 mM in their absence to 0.7 mM in their presence. The Km for phosphocholine was not affected by lipid activators. The activity of CT[delta 257-367] was comparable to the activity of wild-type CT in the absence of lipid activators and the CTP Km for CT[delta 257-367] was 13.9 mM. The enzymatic properties of the CT[delta 231-367] mutant were comparable to those exhibited by the CT[257-367] mutant indicating that removal of residues 231 through 257 did not have any additional influence on the lipid regulation of the enzyme. Thus, the region between residues 257 and 312 was required to confer lipid regulation on CT, and the association of activating lipids with this region of the protein stimulated catalysis by increasing the affinity of the enzyme for CTP.

Animals↗
PubMed1995

Characterization of a 95 kilodalton membrane glycoprotein associated with multi-drug resistance.

Over-expression of a 95-kDa membrane protein (P-95) has been reported previously in the multi-drug-resistant (MDR) breast cancer cell line MCF-7/AdrVp and the MDR small cell lung cancer line NCI-H1688. We have now developed anti-sera against gel-purified }-95 protein from each of these cell lines. Western blotting with each serum demonstrates a broad band at 95-kDa with detergent-solubilized membrane proteins from MCF-7/AdrVp and NCI-H1688 cells, which is barely detectable in membrane proteins from drug-sensitive parental MCF-7 cells. Each anti-serum cross-reacts with a 190-kDa membrane protein (P-190) in NCI-H1688 but not MCF-7/AdrVp cells. Immunoblotting and silver staining of NCI-H1688 membrane proteins separated by two-dimensional gel electrophoresis demonstrates that P-95 and P-190 run as broad streaks with low iso-electric points. Incubation of NCI-H1688 cells with tunicamycin or cleavage of carbohydrate residues of NCI-H1688 or MCF-7/AdrVp membrane proteins with PNGase F leads to the appearance of a sharp 35-kDa band reactive with anti-P-95 antisera. This 35-kDa protein has been isolated by two-dimensional gel electrophoresis. Neuraminidase digestion converts P-95 to a broad 65-kDa immunoreactive band, indicating the presence of terminal sialic acid residues. In conclusion, P-95 is an N-linked sialoglycoprotein with a 35-kDa polypeptide core.

Blotting, Western↗
PubMed1995

An SH3-binding site conserved in Bruton's tyrosine kinase and related tyrosine kinases mediates specific protein interactions in vitro and in vivo.

Mutations in Bruton's tyrosine kinase (Btk) have been associated with immunodeficiencies in man and in the mouse. Btk and two related proteins, Itk and Tec, are members of a distinct family of tyrosine kinases. These kinases are believed to function in various receptor-mediated signaling pathways, but their specific functions are as yet undefined. Btk and its homologues share extensive sequence similarity, including a conserved region, the Tec-homology (TH) domain, that has been proposed to mediate specific intermolecular or intramolecular interactions. The TH region of Btk contains a functional SH3-binding site at residues 189-192. SH3 binding is selective: Btk is retained by the SH3 domain of Fyn but not by that of Blk, another Src-type kinase. TH-SH3 binding in vitro is abolished by specific, single amino acid substitutions within the Btk TH domain or the Fyn SH3 domain. We provide two lines of evidence that the SH3-binding site in the Btk TH domain mediates protein interactions in intact cells. First, treatment of cells with pervanadate induces an increase in the phosphotyrosine content of kinase-inactive Btk; this response is substantially reduced by a mutation that inactivates the SH3-binding site in the Btk TH domain. Second, in cell lysates Btk is found in association with an as yet unidentified 72-kDa phosphotyrosine-containing protein; this interaction requires a functional SH3-binding site in the TH domain. The TH domain may therefore interact in vivo with other proteins that regulate the phosphorylation state of Btk.

Agammaglobulinaemia Tyrosine Kinase↗
PubMed1995

Thrombocytopenia in dogs induced by granulocyte-macrophage colony-stimulating factor: increased destruction of circulating platelets.

Administration of recombinant canine granulocyte-macrophage colony-stimulating factor (rcGM-CSF) to normal dogs in previous studies induced an increase in peripheral blood neutrophils and a dose-dependent decrease in platelet counts. In six dogs that received the highest tested dose of rcGM-CSF (50 micrograms/kg/d) for a minimum of 12 days, the mean nadir of the platelet count was 46,000/microL (range, 4,000 to 91,000/microL) on day 9 +/- 1.1 after starting therapy, compared with a mean baseline platelet count of 398,000/microL (range, 240,000 to 555,000/microL). In three dogs, survival of autologous 111In-labeled platelets was reduced from a mean of 4.9 days to 1.3 days during the administration of rcGM-CSF. Biodistribution studies with gamma camera imaging indicated that there was an increase in mean hepatic uptake during the administration of rcGM-CSF, from 15% to 44% of the total injected 111In-labeled platelets at 2 hours, whereas splenic uptake was not significantly changed. In contrast, in two evaluable dogs who were recipients of 111In-labeled platelets from matched allogeneic donors receiving rcGM-CSF, platelet survival was not reduced and no increased hepatic uptake was noted. A third dog became alloimmunized to the matched donor platelets and was not evaluable. Immunohistologic studies of liver and spleen were performed with monoclonal antibodies specific for canine gpIIb/IIIa and P-selectin in dogs treated with rcGM-CSF and compared with untreated controls. On treatment, a marked reduction of platelets in the red pulp of the spleen was evident, and in general, the presence of platelet antigen in the liver was unchanged. Therefore, platelets were not being sequestered, but destroyed in the liver and spleen. The platelet antigens, P-selectin and gpIIb/IIIa, were identified in association with Kupffer cells in the liver, but no difference in the number of distribution of these Kupffer cells was found between controls and rcGM-CSF-treated dogs. In the spleen during rcGM-CSF treatment, most platelet antigens were associated with large mononuclear cells in the marginal zone. During administration of rcGM-CSF, CD1c and CD11c expression was increased on Kupffer cells. Platelet P-selectin expression and binding of leukocytes to circulating platelets were unchanged from baseline studies with rcGM-CSF treatment. In conclusion, during the administration of rcGM-CSF to dogs, a local process in the liver and spleen is induced resulting in thrombocytopenia.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗
PubMed1995

Studies on the distribution of vasopressin-immunoreactive neuronal perikarya and their fibers in the hypothalamus of Tupaia belangeri.

The distribution of vasopressin (VP)-immunoreactive neuronal perikarya and its fibers had been studied in the hypothalamus of Tupaia belangeri using the avidin-biotin complex (ABC) immunocytochemical technique. VP-immunoreactive neurons were found in the hypothalamic paraventricular nucleus (PVN), supraoptic nucleus (SON), accessory supraoptic nucleus (ASN), hypothalamic lateral nucleus (HLN), perifornical nucleus (PFN) and ansa peduncularis (AP) but not in the suprachiasmatic nucleus (SCN). VP neurons of the rostral PVN could be divided into three subnuclei and the caudal PVN could also be divided although not so distinctly into four subnuclei. The VP-immunoreactive neuronal perikarya of SON were divided into three parts, i.e., medioventral, mediodorsal and laterodorsal. Three types of VP neuronal perikarya, i.e., large, medium and small cells, existed in PVN and SON. Between PVN and SON, there were a large number of VP immunopositive nerve fibers. In addition, there were numerous immunopositive fibers projecting into the infundibulum and the neurohypophysis. VP-immunoreactive-positive products localized in the large granular vesicles and on the rough-surfaced endoplasmic reticulum could be seen under electron microscope.

Animals↗
PubMed1995

Crystal structure of the site-specific recombinase gamma delta resolvase complexed with a 34 bp cleavage site.

The structure of gamma delta resolvase complexed with a 34 bp substrate DNA has been determined at 3.0 A resolution. The DNA is sharply bent by 60 degrees toward the major groove and away from the resolvase catalytic domains at the recombination crossover point. The C-terminal one third of resolvase, which was disordered in the absence of DNA, forms an arm and a 3-helix DNA-binding domain on the opposite side of the DNA from the N-terminal domain. The arms wrap around the minor groove of the central 16 bp, and the DNA-binding domains interact with the major grooves near the outer boundaries of the binding site. The resolvase dimer is asymmetric, particularly in the arm region, implying a conformational adaptability that may be important for resolvase binding to different DNA sites in the synaptosome. It also raises the possibility of a sequential single-strand cleavage mechanism.

Amino Acid Sequence↗
PubMed1995

Antibodies to Schistosoma japonicum (Asian bloodfluke) paramyosin induced by nucleic acid vaccination.

Nucleic acid vaccination by intramuscular or intradermal delivery of DNA plasmids encoding antigenic proteins has been shown to confer protection in experimental animals against viruses and unicellular protozoan parasites. However, this revolutionary approach has not been tested for induction of immunity to multicellular parasites, such as trematode worms. We report here, for the first time, that murine antibodies can be induced by intramuscular injection with plasmid DNA encoding fragments of Schistosoma japonicum paramyosin (Sj97), a 97 kDa molecule and a promising vaccine candidate in schistosomiasis. An additional construct containing the gene encoding full-length glutathione S-transferase (Sj26), another recognised anti-schistosome vaccine target, failed to raise detectable levels of specific antibody.

Animals↗
PubMed1995

Lipid activation of CTP:phosphocholine cytidylyltransferase is regulated by the phosphorylated carboxyl-terminal domain.

The role of the phosphorylated carboxyl-terminal domain of CTP:phosphocholine cytidylyltransferase (CT) in the regulation of enzyme activity was investigated by comparing the catalytic properties of wild-type CT to two mutant proteins with altered carboxyl-terminal phosphorylation domains. CT isolated from a baculovirus expression system was extensively phosphorylated at multiple sites in the carboxyl-terminal domain. The CT[S315A] mutant lacked a major CT phosphorylation site, and the carboxyl-terminal deletion mutant, CT[delta 312-367], was not phosphorylated. The higher activities of CT[delta 312-367] and CT[S315A] relative to CT were attributed to differences in the sensitivities of the enzymes to lipid activators. The rank order of the apparent Km values for activation by either phosphatidylcholine/oleic acid or phosphatidylcholine/diacylglycerol was CT > CT[S315A] > CT[delta 312-367]. In addition, CT exhibited negative cooperativity in its activation by phosphatidylcholine/oleic acid (nH = 0.64) and phosphatidylcholine/diacylglycerol (nH = 0.74) vesicles, whereas CT[delta 312-367] and CT[S315A] did not. These data support the concept that the phosphorylation of the CT carboxyl-terminal domain interferes with the activation of CT by lipid regulators.

Animals↗