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Y Chu

Publications and source records attributed to Y Chu.

At least 91 records · Page 5Linked to original sources
PubMed1997

Altered vascular function after adenovirus-mediated overexpression of endothelial nitric oxide synthase.

Gene transfer with replication-deficient adenovirus is a potentially useful tool to study vascular biology. We have constructed a replication-deficient adenovirus (AdRSVeNOS) that carries cDNA for endothelial nitric oxide synthase (eNOS). Transfection of COS-1 cells with AdRSVeNOS increased nitric oxide synthase activity (measured as production of L-citrulline from L-arginine) that was calcium dependent and inhibited by N omega-nitro-L-arginine methyl ester. To investigate effects of overexpression of eNOS on vascular function, we incubated common carotid arteries from rabbits in organ culture with AdRSVeNOS or AdRSV beta gal encoding beta-galactosidase. Transgene expression and responses to vasoactive agents were examined 1 day after transduction. Histochemical staining of beta-galactosidase and immunohistochemistry for eNOS indicated transgene expression in endothelium and adventitial cells. After precontraction with phenylephrine, vessels treated with AdRSVeNOS demonstrated greater relaxation to acetylcholine than vessels treated with vehicle or AdRSV beta gal. Relaxation to calcium ionophore A-23187 was much greater in vessels treated with AdRSVeNOS than in vessels treated with vehicle or AdRSV beta gal. Augmented relaxation in response to A-23187 was also observed after denudation of endothelium in vessels treated with AdRSVeNOS and was inhibited by N omega-nitro-L-arginine. Thus vasorelaxation in response to stimuli that release nitric oxide is augmented after adenovirus-mediated overexpression of eNOS. Transgene expression in adventitial cells appears to be sufficient to alter vasomotor function.

Adenoviruses, Human↗
PubMed1997

Augmented adenovirus-mediated gene transfer to atherosclerotic vessels.

Vascular endothelium is an important target for gene transfer in atherosclerosis. In this study, we examined gene transfer to normal and atherosclerotic blood vessels from two species, using an organ culture method. Using normal aorta, we determined optimal dose, duration of exposure to adenovirus, and duration of incubation of vessels in tissue culture. Aortas from normal and atherosclerotic monkeys were cut into rings and incubated for 2 hours with a recombinant adenovirus, carrying the reporter gene for beta-galactosidase driven by a cytomegalovirus (CMV) promoter. After 20 hours of incubation, transgene expression was assessed with a morphometric method after histochemical staining and a chemiluminescent assay of enzyme activity. Expression of beta-galactosidase after histochemical staining, expressed as percentage of total cells, was similar in adventitial cells of normal monkeys (21 +/- 4%, mean +/- SE%) and atherosclerotic monkeys (25 +/- 12%). Transgene expression in endothelium was higher in atherosclerotic than in normal vessel (53 +/- 3% versus 27 +/- 7%, P < .05). Chemiluminescent assay indicated greater beta-galactosidase activity (2.5 +/- 0.6 mU/mg of protein) in the intima and media of atherosclerotic than normal vessels (0.6 +/- 0.2 mU/mg of protein, P < .05). Aortas from normal (n = 6) and atherosclerotic (n = 5) rabbits also were examined. Transgene expression (after histochemical staining) in endothelium was much greater in atherosclerotic than normal rabbits (39 +/- 3% versus 9 +/- 2%, P < .05) and expression in adventitial cells was similar (normal 23 +/- 2%, atherosclerotic 24 +/- 4%). Chemiluminescent assay indicated greater beta-galactosidase activity (1.2 +/- 0.4 mU/mg of protein) in the intima and media of atherosclerotic than normal vessels (0.2 +/- 0.1 mU/mg protein, P < .05). These findings suggest that an adenoviral vector with a CMV promoter provides similar transgene expression in adventitia of both normal and atherosclerotic vessels. Gene transfer to the endothelium was much more effective in atherosclerotic than in normal vessels. Thus it may be possible to achieve greater transgene expression in atherosclerotic than in normal arteries.

Adenoviridae↗
PubMed1997

Increase in TUNEL positive cells in aorta from diabetic rats.

TUNEL staining, which allows detection of fragmented DNA in situ, is commonly used as an indication of apoptosis. Recent studies suggest that apoptosis is increased in several pathophysiological conditions. In this study we examined the hypothesis that chronic diabetes is associated with an increase in TUNEL staining of the aorta. Diabetic rats were studied 4-5 months after injection of streptozotocin (50 mg/kg). Aorta of diabetic and control rats were examined for TUNEL staining, morphology by electron microscopy, and DNA contamination in RNA preparation by polymerase chain reaction (PCR). TUNEL staining of aortic sections showed a 6 fold increase of positive cells in the media of diabetic aorta (22 +/- 6%) (mean +/- SE) compared with aorta from age-matched controls (3.6 +/- 0.9%, p < 0.05). Electron microscopy demonstrated typical apoptotic cells and bodies in the media of aorta from diabetic but not control rats. DNA contamination was found in RNA prepared from diabetic aorta, which was detected using PCR, which is consistent with increased DNA fragmentation. Increased TUNEL staining was not observed in rats with hyperglycemia 3 days after injection of streptozotocin. In conclusion, severe chronic diabetes is associated with an increase in TUNEL staining, and perhaps apoptosis, in the aorta. We speculate that increased apoptosis may compensate for increased proliferative activity in diabetic blood vessels.

Animals↗
PubMed1997

[Colony-stimulating activities of serum from patients with severe aplastic anemia].

OBJECTIVE: To investigate the correlation between colony-stimulating activities of serum from patients with severe aplastic anemia (SAA) and their responses to immunosuppressive therapy (IST). METHODS: In vitro effects of sera from SAA patients before and after IST and from normal subjects on human marrow colony growth were examined in a semisolid methylcellulose culture system for burst promoting activity (BPA) and granulocyte/macrophage colony-stimulating activity (GM-CSA). Serum erythropoietin (Epo) level was also measured by ELISA method before and after IST in SAA patients. RESULTS: Serum BPAs in SAA patients at diagnosis were significantly higher than that in normal controls (P<0.001), but serum GM-CSAs in 13 of 22 SAA patients were normal,and in 9 of 22 extremely low as compared with normal values. After IST, serum BPA and GM-CSA in SAA patients had no changes. Serum Epo levels in SAA patients at diagnosis were significantly higher than that in normal controls (P<0.001). After IST,serum Epo levels declined in responded patients, while further increased in nonresponded. CONCLUSION: Serum GM-CSA of SAA patients was a predictive factor for responsiveness to IST, and a normal value was associated with a good response.

Anemia, Aplastic↗
PubMed1997

[Correlation between the serum levels of interleukin-2 and soluble interleukin-2 receptor and the response to immunosuppressive therapy in severe aplastic anemia patients].

OBJECTIVE: To explore factors influencing the effectiveness of immunosuppressive therapy in severe aplastic anemia (SAA). METHODS: The serum levels of interleukin-2 (IL-2) and soluble IL-2 receptor (sIL-2R) of 22 SAA patients and 15 normal controls were measured by immunoenzyme assay. RESULTS: Comparing with the normal controls, 40.9% and 63.6% of the SAA patients had higher serum levels of IL-2 (>840ng/L) and of sIL 2R (>320 x 10(3) U/L), respectively. Those who had higher IL-2 levels and higher sIL-2R levels got better responses to cyclosporin A(CsA) and CsA plus antilymphocyte globulin (ALG). CONCLUSION: The serum levels of IL-2 and sIL-2R were correlated with the responsiveness to immunosuppressive therapy in SAA patients and might be useful for directing treatment.

Adolescent↗
PubMed1997

[HLA-DR-positive T lymphocytes in the bone marrow and peripheral blood of patients with severe aplastic anemia and their in vitro inhibitory effecton BFU-E and CFU-GM].

OBJECTIVE: [corrected] To elucidate the role of immunodysfunction in the pathogenesis of severe aplastic anemia (SAA). METHODS: HLA-DR+ T cells were detected in bone marrow (BM) and peripheral blood (PB) of 20 SAA patients and controls (9 for BM and 11 for PB). PHA-LYCM conditioned medium was prepared, and its in vitro effect on normal bone marrow BFU-E and CFU-GM was studied. RESULTS: The percentages of HLA-DR+ T lymphocytes in BM and PB of newly diagnosed SAA patients were significantly higher than that of controls (P<0.001), and that of recovered SAA (rSAA) patients after ALG therapy (P<0.05). Compared with controls, the PBA-LYCMs from newly diagnosed SAA patients showed significant inhibitory effect on normal BFU-E and CFU-GM (P<0.001). CONCLUSION: The activated T lymphocyte may play an important role in the pathogenesis of SAA.

Adolescent↗
PubMed1997

[Immunophenotyping study on the blood cells of the patients with paroxysmal nocturnal hemoglobinuria].

OBJECTIVE: To detect the expression of glycosyl-phosphatidyl-inositol (GPI) anchored protein on the blood cell membrane and its implication in the diagnosis of paroxysmal nocturnal hemoglobinuria (PNH). METHODS: The GPI anchored protein was detected by indirect immunofluorescence method with a panel of monoclonal antibodies (CD55, CD59, CD16, CD58, HIR2, CD15, CD2) in 22 PNH patients. RESULTS: The abnormal granulocytes and erythrocytes with deficient GPI anchored proteins were identified in all PNH patients whether the routine hemolysis tests for PNH, including Ham test and mCLST, were positive or negative. However, lymphocytes with PNH-defect were found in only 14 patients. CONCLUSION: GPI anchored protein assay provided sensitive and specific method for the diagnosis of PNH.

Adult↗
PubMed1997

[Study on the immunophenotype of myeloid cells in myelodysplastic syndromes and its clinical implications].

OBJECTIVE: To explore the immunophenotype of myeloid cells in myelodysplastic syndromes and its clinical implications. METHODS: A panel of monoclonal antibodies was used to detect CD13, CD33, CD15 and CD14 on the membrane surfaces of myeloid cells in the bone marrow from 51 patients with myelodysplastic syndromes (MDS), 21 with aplastic anemia (AA), 21 with paroxysmal nocturnal hemoglobinuria (PNH), and 15 normal subjects, by immunoenzymatic assay. The morphology and chromosome karyotype of bone marrow cells of MDS patients were also examined. RESULTS: CD14+ cells, CD13+ cells and CD33+ cells in the bone marrow were more in MDS patients than in normal controls, AA patients and PNH patients. CD15+ cells in the bone marrow were less in MDS patients than in normal controls. The percentages of CD14, CD13 and CD33 positive cells in the bone marrow of MDS patients were related to the percentage of myeloblast, the chromosomal aberrations and the response to treatment. CONCLUSION: There was an immunophenotypic misexpression of myeloid cells in MDS patients. Immunophenotype analysis of myeloid cells might be useful for the diagnosis and directing treatment in MDS patients.

Adolescent↗
PubMed1997

[Colony formation of bone marrow hematopoietic progenitor cells in patients with severe aplastic anemia and its relation to the results of immunosuppressive therapy].

OBJECTIVE: To explore the defect of hematopoietic stem cell and analyze the relationship between the colony formation capacity of bone marrow hematopoietic progenitor cells and the results of immunosuppressive therapy (IST) in severe aplastic anemia (SAA) patients. METHODS: Methylcellulose semisolid culture was used. RESULTS: Thirty patients with SAA at diagnosis were studied. In 90% of the patients, the CFU-E and CFU-GM yields were strikingly decreased and in 56.7% of the patients there was no colony formation at all. Nevertheless, there was still 10% of the patients having normal CFU-E, BFU-E or/ and CFU-GM yields. After IST, 59.1% of the patients showed colony formation improvement. The difference between the results obtained pre- and post-IST(for CFU-E and BFU-E, P < 0.01, for CFU-GM, P < 0.05) was significant. In the majority of the patients, the colony formation capacities were still under the normal post-IST. Moreover, 22.7% of the patients remained no colony formation post-IST. The IST response rates between the patients with or without colony formation had no statistically difference (80% versus 50%, P < 0.05). The increment of colony formation appeared later than the therapeutic effect did, but frequently concurred with the appearance of magkaryocytes in the bone marrow smear. CONCLUSION: In the majority of SAA patients, the hematopoietic stem cells or progenitors were defective perhaps caused by immune damage; IST can improve the hematopoiesis in vitro and in vivo of SAA patients.

Adolescent↗
PubMed1996

Effect of activation of protein phosphatase 1 on sulfhydryl reactivity.

Myofibril protein phosphatase 1 (PP1) from bovine heart, identified as PP1alpha, was purified in a latent form which was dependent on Co2+ or Mn2+ for activity (Y. Chu, S. E. Wilson, and K. K. Schlender (1994) Biochim. Biophys. Acta 1208, 45-54). This was also true for recombinant PP1 alpha expressed in Escherichia coli (Z. Zhang, G. Bai, S. Deans-Zirattu, M. F. Browner, and E. Y. C. Lee (1992) J. Biol. Chem. 267, 1484-1490). Here we report on the change in the sulfhydryl reactivity during the cation activation process. The activation of myofibrillar PP1 by Co2+ was prevented by 10 mM dithiothreitol (DTT) and incubation of the Co2+-activated enzyme with 50 mM DTT reversed the activation. Activation of recombinant PP1alpha was associated with 57Co2+ incorporation into PP1. DTT reversal of Co2+-activated PP1 was accompanied by release of Co2+ from the enzyme. The latent PP1 modified with 2-nitro-5-thiocyanobenzoic acid (NTCB) or N-ethylmaleimide (NEM) did not bind Co2+ and could not be activated by Co2+. Conversely, the Co2+-activated PP1 was resistant to inactivation with NTCB and less sensitive to NEM. Similarly, PP1 pretreated with NTCB was not activated by Mn2+ and the Mn2+-activated enzyme was also resistant to NTCB inhibition. The number of sulfhydryls of nondenatured PP1, reactive with 5, 5'-dithiobis[2-nitrobenzoic acid] (DTNB), was reduced from approximately 8 to 2-3 mol/mol when the enzyme was activated with Co2+ or Mn2+. After denaturation with guanidine-HCl, the number of reactive sulfhydryls of nonactivated PP1 and Co2+-activated PP1 was approximately 10 mol/mol enzyme. These results suggest that when PP1 is activated by Co2+ or Mn2+, the enzyme undergoes a conformational change resulting in some of the cysteine sulfhydryls no longer being accessible to chemical modification.

Animals↗
PubMed1996

The mitogen-activated protein kinase phosphatases PAC1, MKP-1, and MKP-2 have unique substrate specificities and reduced activity in vivo toward the ERK2 sevenmaker mutation.

Mitogen-activated protein (MAP) kinases can be grouped into three structural families, ERK, JNK, and p38, which are thought to carry out unique functions within cells. We demonstrate that ERK, JNK, and p38 are activated by distinct combinations of stimuli in T cells that simulate full or partial activation through the T cell receptor. These kinases are regulated by reversible phosphorylation on Tyr and Thr, and the dual specific phosphatases PAC1 and MKP-1 previously have been implicated in the in vivo inactivation of ERK or of ERK and JNK, respectively. Here we characterize a new MAP kinase phosphatase, MKP-2, that is induced in human peripheral blood T cells with phorbol 12-myristate 13-acetate and is expressed in a variety of nonhematopoietic tissues as well. We show that the in vivo substrate specificities of individual phosphatases are unique. PAC1, MKP-2, and MKP-1 recognize ERK and p38, ERK and JNK, and ERK, p38, and JNK, respectively. Thus, individual MAP kinase phosphatases can differentially regulate the potential for cross-talk between the various MAP kinase pathways. A hyperactive allele of ERK2 (D319N), analogous to the Drosophila sevenmaker gain-of-function mutation, has significantly reduced sensitivity to all three MAP kinase phosphatases in vivo.

Animals↗
PubMed1996

Activation of protein phosphatase 1. Formation of a metalloenzyme.

The recombinant catalytic subunit of protein phosphatase 1 is produced as an inactive enzyme which can be activated by Mn2+ (Zhang, Z., Bai, G., Deans-Zirattu, S., Browner, M. F., and Lee, E. Y. C. (1992) J. Biol. Chem. 267, 1484-1490). In this report, we have investigated the effects of divalent cations on the activity of recombinant catalytic subunit of protein phosphatase 1. Latent phosphatase 1 can be activated by Co2+ or Mn2+, whereas other metal ions tested including Fe2+, Zn2+, Mg2+, Ca2+, Cu2+, or Ni2+ were not effective or were only weakly effective in activating the enzyme. The Mn(2+)-stimulated activity was susceptible to inactivation by EDTA; however, the Co(2+)-activated phosphatase was stable after dilution and chelation of the Co2+ with excess EDTA. After stable activation of phosphatase 1 using 57Co2+, a stoichiometric amount of 57Co2+ was shown to be tightly bound to phosphatase 1. These findings demonstrate for the first time the generation of a stable metalloenzyme form of phosphatase 1. Fe2+ reversibly deactivated the Co(2+)-stimulated activity, but did not displace the bound Co2+. Interestingly, treatment of the enzyme with a combination of Fe2+ and Zn2+ (but not the individual metal ions) significantly activated phosphatase 1. These results suggest that at least two metal binding sites exist on the enzyme and that protein phosphatase 1 may be an iron/zinc metalloprotein in vivo.

Enzyme Activation↗
PubMed1996

Vascular expression of inducible nitric oxide synthase is associated with activation of Ca(++)-dependent K+ channels.

We tested the hypothesis that expression of inducible nitric oxide synthase (NO-synthase) in response to endotoxin (lipopolysaccharide) produces activation of potassium channels. Contraction of the rat thoracic aorta in response to phenylephrine was measured in vitro after treatment in vivo for 15 hr with vehicle (control) or lipopolysaccharide (10 mg/kg i.p.). Impaired contraction in response to phenylephrine was used as an index of inducible NO-synthase expression, and activation of potassium channels was examined with specific inhibitors. Contraction in response to 10(-5) M phenylephrine (expressed as a percentage of contraction in response to 85 mM KCI) was markedly impaired in lipopolysaccharide-treated rats, compared with control (15 +/- 5% vs. 131 +/- 10%, P < .05, mean +/- S.E.). Expression of inducible NO-synthase mRNA in the vessel wall in lipopolysaccharide-treated rats was confirmed using reverse transcription-polymerase chain reaction. Contraction of the aorta in lipopolysaccharide-treated rats was restored to normal by 0.3 mM aminoguanidine (an inhibitor of inducible NO-synthase). Contraction of the aorta in response to phenylephrine, which was inhibited by lipopolysaccharide, was not affected by glibenclamide (an inhibitor of ATP-sensitive potassium channels) but was increased 2-fold (P < .05) by iberiotoxin (50 nM), an inhibitor of Ca(+2)-dependent potassium channels. Relaxation of the aorta in response to sodium nitroprusside, an exogenous donor of nitric oxide, and 8-bromo-cyclic GMP was also inhibited by iberiotoxin. These findings suggest that nitric oxide produced by vascular expression of inducible NO-synthase activates calcium-dependent potassium channels and that this mechanism may contribute to impaired vasoconstrictor responses during sepsis.

Animals↗
PubMed1996

Studies and applications of NiTi shape memory alloys in the medical field in China.

The biomedical study of NiTi shape memory alloys has been undertaken in China since 1978. A series of stimulating corrosion tests, histological observations, toxicity tests, carcinogenicity tests, trace nickel elements analysis and a number of clinical trials have been conducted. The results showed that the NiTi shape memory alloy is a good biomaterial with good biocompatibility and no obvious local tissue reaction, carcinogenesis or erosion of implants were found experimentally or clinically. In 1981, on the basis of fundamental studies, a shape memory staple was used for the first time inside the human body. Subsequently, various shape memory devices were designed and applied clinically for internal fixation of fractures, spine surgery, endoprostheses, gynaecological and craniofacial surgery. Since 1990, a series of internal stents have been developed for the management of biliary, tracheal and esophageal strictures and urethrostenosis as well as vascular obturator for tumour management. Several thousand cases have been treated and had a 1-10 year follow-up and good clinical results with a rather low complication rate were obtained.

Adult↗
PubMed1996

[Experience with reoperation on mitral valve prostheses].

Heart valve replacement after closed mitral commissurotomy and mitral valve replacement with bioprostheses may result in complication and higher mortality and morbidity. Mitral valve replacement was performed in 32 cases after closed mitral commissurotomy and 12 cases after mitral valve replacement with bioprostheses from April 1986 to May 1994 in our hospital. The eraly mortality was 6.8% (3/42). The choice of operation and methods, myocardial preservation, measurement of heart resuscitation, and prevention of perioperative hemorrhage were discussed.

Adult↗
PubMed1996

[Effects of AT-I on murine cytokins productions and NK cell activity in vivo].

Antitumor-I (AT-I) is a traditional Chinese medicine, which known to be combined with chemotherapeutic agent on antitumor effects. In this paper, the effections of AT-I on interleukin-2 (IL-2) and interleukin-6 (IL-6) was detected and NK cell activity were observed in normal and tumor-bearing BALB/c mice in vivo. The results showed that AT-I could enhance IL-2 and IL-6 production in normal mice, and IL-2 product in tumor-bearing mice. The results suggest that AT-I at a appropriate dose could modulate immunofunctions, so known as a biological response modifier (BRM).

Animals↗