Biomedical subjects

J Jin

Publications and source records attributed to J Jin.

At least 109 records · Page 6Linked to original sources

PubMed1998

Molecular basis for ADP-induced platelet activation. II. The P2Y1 receptor mediates ADP-induced intracellular calcium mobilization and shape change in platelets.

ADP is an important platelet agonist causing shape change from smooth discoid shape to spiculated spheres and platelet aggregation. However, the molecular mechanisms involved in ADP-induced platelet activation have not been elucidated. We demonstrated earlier the existence of two distinct ADP receptors on platelets, one coupled to phospholipase C, P2TPLC, and the other to inhibition of adenylyl cyclase, P2TAC (Daniel, J. L., Dangelmaier, C., Jin, J., Ashby, B., Smith, J. B., and Kunapuli, S. P. (1998) J. Biol. Chem. 273, 2024-2029), in addition to the previously described P2X1 receptor. Here we report the cloning of a cDNA clone encoding the P2Y1 receptor from a human platelet cDNA library by homology screening with radiolabeled P2Y1-P2Y6 receptor cDNAs. ADP or 2-methyl(thio)-ADP-induced intracellular calcium increases were inhibited by the P2Y1 receptor-specific antagonists, adenosine 3'-phosphate 5'-phosphosulfate (A3P5PS), adenosine 3'-phosphate 5'-phosphate (A3P5P), and adenosine 2'-phosphate 5'-phosphate (A2P5P), in a concentration-dependent manner, but not by ARL 66096 or alpha, beta-MeATP. A3P5PS, A3P5P, and A2P5P also inhibited the shape change of aspirinated platelets induced by 10 microM ADP or 3 microM 2-methyl-(thio)-ADP in a concentration-dependent manner, with complete inhibition occurring at 300 microM. On the other hand ARL 66096 (100 nM), a potent P2TAC antagonist and alpha, beta-methylene-ATP (40 microM), a P2X1 receptor agonist, had no effect on ADP-induced platelet shape change. On the contrary, ADP-induced inhibition of adenylyl cyclase was blocked by ARL 66096, but not by alpha, beta-MeATP or the P2Y1 receptor-specific antagonists, A3P5PS, A3P5P, or A2P5P. These results demonstrate the role of the P2Y1 receptor in ADP-induced platelet shape change and calcium mobilization and support the idea that several P2 receptors are involved in the regulation of different aspects of platelet stimulus-response coupling.

Adenosine Diphosphate↗

PubMed1998

C-fos mediates cocaine inhibition of NGF-induced PC12 cell differentiation.

In utero cocaine exposure can affect CNS development. Previous studies showed that cocaine inhibits neuronal differentiation in a dose-dependent fashion, in nerve growth factor (NGF)-stimulated PC12 cells, without affecting cell viability. NGF activates intracellular signaling proteins, specific immediate-early genes (IEG) including a transient peak of c-fos expression, and induction of late genes expression, leading to the neuronal phenotype. We hypothesized that cocaine interferes with NGF signaling. Therefore, we examined the pattern of c-fos expression in our cellular model. Time course of c-fos expression up to 72 h was determined in cells treated with NGF 20 ng/ml and cocaine 10 microgram/ml (a moderately toxic level) by RT-PCR analysis. Total RNA was isolated from cells, and levels of c-fos mRNA were estimated using gene-specific primers. In both control and experimental conditions, c-fos level was maximal at 0.5 h. In the control cells, c-fos expression declined rapidly to less than 5% of the 0.5h value, while in the cocaine-treated cells, c-fos level persisted through the 72-h exposure. Adding c-fos antisense to cells treated with NGF and cocaine resulted in significant improvement of neurite out-growth, from 28% (NGF + cocaine) to 89% (NGF + cocaine + c-fos antisense) of control differentiation after 72 h of exposure (Dunnet's T < 3.24). Inhibitory effects of cocaine on NGF-induced PC12 differentiation may be attributed to alteration of c-fos expression. Further studies will be required to examine the role of D1 receptor activation in mediating c-fos expression and to explore the effects of cocaine on other IEGs.

Animals↗

PubMed1998

Cardiokymograph system with a capacitance transducer and its preliminary application in the measurement of heart wall movement.

A novel cardiokymograph system is introduced. The new system features a capacitance transducer with increased sensitivity and can be used in multichannel measurements. The novelty of this technique is the injection of a current into the patient coupled with the use of a capacitive displacement transducer and the possibility of multichannel monitoring. It provides for the possibility of removing breath noise when some signal processing technique, such as adaptive filtering, is used. Further investigation is needed to demonstrate clinical significance and pathologies.

Adult↗

PubMed1998

Distribution of P2Y receptor subtypes on haematopoietic cells.

1. RT-PCR-southern hybridization analyses with radiolabelled P2Y receptor cDNAs as probes indicated that the peripheral blood leukocytes and the human umbilical vein endothelial cells express P2Y1, P2Y2, P2Y4 and P2Y6 receptors. 2. Of the haematopoietic cell lines tested, promonocytic U937 cells express P2Y2 and P2Y6, but not P2Y1 or P2Y4; promyelocytic HL-60 cells express the P2Y1, P2Y2 and P2Y6 receptors but not the P2Y4 receptor; K562 cells express P2Y1 but not P2Y2, P2Y4 or P2Y6; and Dami cells express P2Y1, P2Y2, P2Y4 and P2Y6 receptors. 3. Of the peripheral blood leukocytes tested, polymorphonuclear cells express P2Y4 and P2Y6 but not P2Y1 or P2Y2 receptors; monocytes express P2Y1, P2Y2, P2Y4 and P2Y6 receptors and lymphocytes express P2Y1, P2Y2, P2Y4 and P2Y6 receptors. 4. These results suggest a physiological role for different P2Y receptor subtypes in the extracellular nucleotide-mediated stimulation of monocytes, neutrophils, lymphocytes and endothelial cells.

Blotting, Southern↗

PubMed1998

A virus-encoded RNA polymerase purified from baculovirus-infected cells.

A DNA-dependent RNA polymerase was purified to homogeneity, starting from insect cells infected with the baculovirus Autographa californica nuclear polyhedrosis virus (AcNPV). The purified polymerase supported accurate and specific transcription from late and very late promoters but was not active on viral early promoters. Thus, promoter recognition is an integral function of the purified enzyme. The purified RNA polymerase was composed of only four equimolar subunits, which makes it the simplest DNA-directed RNA polymerase from a eukaryotic source described so far. Amino-terminal protein sequencing, peptide fingerprinting, and immunochemical analyses were used to identify the four subunits, all of which are virus encoded. Overexpression of the four viral proteins (LEF-8, LEF-4, LEF-9, and p47) in baculovirus-infected cells resulted in a significant increase in the levels of RNA polymerase produced in the infected cells. Thus, the overexpression data are consistent with our identification of the RNA polymerase subunits.

Amino Acid Sequence↗

PubMed1998

Guanylyltransferase activity of the LEF-4 subunit of baculovirus RNA polymerase.

The baculovirus Autographa californica nuclear polyhedrosis virus encodes a DNA-dependent RNA polymerase that transcribes viral late genes. This polymerase is composed of four equimolar subunits, LEF-4, LEF-8, LEF-9, and p47. Here we present data indicating that the LEF-4 subunit of RNA polymerase is a guanylyltransferase. Incubation of RNA polymerase in the presence of divalent cation and radiolabeled GTP resulted in the formation of a covalent enzyme-guanylate complex that comigrated with the LEF-4 subunit. The label transfer assay showed an absolute requirement for divalent cation which could be satisfied by either manganese or magnesium. The reaction was specific for guanine nucleotides, and GTP was more effective than dGTP in the formation of enzyme-guanylate complex. To demonstrate that LEF-4 was the guanylyltransferase, the single subunit was overexpressed in baculovirus-infected cells. The overexpressed protein was primarily cytosolic, indicating that other proteins in the RNA polymerase complex were responsible for nuclear targeting of LEF-4. LEF-4 alone was able to covalently bind GMP, although less efficiently than viral RNA polymerase.

Amino Acid Sequence↗

PubMed1998

The LEF-4 subunit of baculovirus RNA polymerase has RNA 5'-triphosphatase and ATPase activities.

The baculovirus Autographa californica nuclear polyhedrosis virus encodes a DNA-dependent RNA polymerase that is required for transcription of viral late genes. This polymerase is composed of four equimolar subunits, LEF-8, LEF-4, LEF-9, and p47. The LEF-4 subunit has guanylyltransferase activity, suggesting that baculoviruses may encode a full complement of capping enzymes. Here we show that LEF-4 is a bifunctional enzyme that hydrolyzes the gamma phosphates of triphosphate-terminated RNA and also hydrolyzes ATP and GTP to the respective diphosphate forms. Alanine substitution of five residues previously shown to be essential for vaccinia virus RNA triphosphatase activity inactivated the triphosphatase component of LEF-4 but not the guanylyltransferase domain. Conversely, mutation of the invariant lysine in the guanylyltransferase domain abolished the guanylyltransferase activity without affecting triphosphatase function. We also investigated the effects of substituting phenylalanine for leucine at position 105, a mutation that results in a virus that is temperature sensitive for late gene expression. We found that this mutation had no significant effect on the ATPase or guanylyltransferase activity of LEF-4 but resulted in a modest decrease in RNA triphosphatase activity.

Acid Anhydride Hydrolases↗

PubMed1998

G protein-dependent activation of smooth muscle eNOS via natriuretic peptide clearance receptor.

In gastrointestinal smooth muscle, the neuropeptides vasoactive intestinal peptide (VIP) and pituitary adenylate cyclase-activating polypeptide (PACAP) induce relaxation by interacting with VIP2/PACAP3 receptors coupled via Gs to adenylyl cyclase and with distinct receptors coupled via Gi1 and/or Gi2 to a smooth muscle endothelial nitric oxide synthase (eNOS). The present study identifies the receptor as the single-transmembrane natriuretic peptide clearance receptor (NPR-C). RT-PCR and Northern analysis demonstrated expression of the natriuretic peptide receptors NPR-C and NPR-B but not NPR-A in rabbit gastric muscle cells. In binding studies using 125I-labeled atrial natriuretic peptide (125I-ANP) and 125I-VIP as radioligands, VIP, ANP, and the selective NPR-C ligand cANP(4-23) bound with high affinity to NPR-C. ANP, cANP-(4-23), and VIP initiated identical signaling cascades consisting of Ca2+ influx, activation of eNOS via Gi1 and Gi2, stimulation of cGMP formation, and muscle relaxation. NOS activity and cGMP formation were abolished (93 +/- 3 to 96 +/- 2% inhibition) by nifedipine, pertussis toxin, the NOS inhibitor, NG-nitro-L-arginine, and the antagonists ANP-(1-11) and VIP-(10-28). NOS activity stimulated by all three ligands in muscle membranes was additively inhibited by Gi1 and Gi2 antibodies (82 +/- 2 to 84 +/- 1%). In reconstitution studies, VIP, cANP-(4-23), and guanosine 5'-O-(3-thiotriphosphate) stimulated NOS activity in membranes of COS-1 cells cotransfected with NPR-C and eNOS. The results establish a unique mechanism for G protein-dependent activation of a constitutive NOS expressed in gastrointestinal smooth muscle involving interaction of the relaxant neuropeptides VIP and PACAP with a single-transmembrane natriuretic peptide receptor, NPR-C.

Animals↗

PubMed1998

Immunohistochemical study of protein kinase C in the testes of cattle and pigs.

We investigated the expression of novel protein kinase C (PKC) delta and theta in the testes of pigs and cattle using Western blot and immunohistochemical analysis. PKC delta and theta are recognized in the testes of pigs and cattle by Western blot analysis. We found in immunohistochemical study that PCK delta was localized in the spermatids of seminiferous tubules, but not in the interstitial cells, while PKC theta was recognized only in the interstitial cells of the testes of in both species. These findings suggest that PKC delta and theta play an important role in the development of spermatozoa and the regulation of androgen in the testicular interstitial cells (probably Leydig cells), respectively.

Animals↗

PubMed1998

Radioimmunoassay of human cardiac acidic isoferritin: a new index for hepatic cancer.

OBJECTIVE: To investigate human cardiac acidic isoferritin as a specific index of hepatic cancer. METHODS: Acidic isoferritin was isolated and purified from human heart muscle. A radioimmunoassay for the acidic isoferritin in human serum has been developed, on the equilibrium method. The antiserum was obtained from rabbits immunized with purified acidic isoferritin. The 125I-acidic isoferritin was prepared by the chloramine-T method. The data were processed using the automated smoothed spline function data processing program. RESULTS: The intra- and inter-assay CV of acidic isoferritin RIA were 1.65% and 9.71%, respectively, and the recovery rate was 102%. The antiserum provided a linear response from 7.0 to 369.6 micrograms/L with ED50 of 27.50 micrograms/L. The cross reactivity with AFP, CEA, lactoferrin and transferrin was negligible, and that with ferritin was 1.74%. The serum acidic isoferritin concentration showed a considerable variation in different sex and age groups. The serum acidic isoferritin was measured in liver diseases including hepatic cancer, hepatic cirrhosis and acute and chronic hepatitis. Its sensitivity for diagnosis of hepatic cancer was 73.05%, independent from the severity of hepatic injury. In 8 malignant tumors studied, acidic isoferritin appeared the most valuable in the diagnosis of hepatic cancer, with its positive, negative, false positive and false negative rates all being ideal. CONCLUSIONS: Acidic isoferritin may turn to be a rather specific index of hepatic cancer. Combination of monitoring both acidic isoferritin and AFP would raise the positive detection and specificity in the diagnosis of hepatic cancer.

Biomarkers, Tumor↗

PubMed1998

Enhancement of (-)-stepholidine on protein phosphorylation of a dopamine- and cAMP-regulated phosphoprotein in denervated striatum of oxidopamine-lesioned rats.

AIM: To study effects of (-)-stepholidine (SPD) on the phosphorylation of a dopamine- and cAMP-regulated phosphoprotein (DARPP-32) in the striatum of oxidopamine-lesioned rats. METHODS: The amount of dephospho-DARPP-32 was measured by a back-phosphorylation assay. RESULTS: In the striatum of control rats, SPD per se had no effect on the phosphorylation of DARPP-32, but it antagonized the decrease by 28% of dephospho-DARPP-32 induced by the D1 agonist SK&F-38393. In the denervated striatum of oxidopamine-lesioned rats, SPD decreased the amount of dephospho-DARPP-32 by 44%. The effect of SPD was completely counteracted by the concomitant administration of the D1 antagonist Sch-23390. CONCLUSION: SPD exhibits D1 agonistic action on DARPP-32 phosphorylation in the denervated striatum of oxidopamine-lesioned rats, but it acts as a D1 antagonist in normal striatum.

2,3,4,5-Tetrahydro-7,8-dihydroxy-1-phenyl-1H-3-ben↗

PubMed1998

[Study on MDM2 and p53 gene proteins expression on acute leukemic cells and its correlation with chemotherapeutic efficacy].

OBJECTIVE: To explore MDM2 and p53 gene proteins expression on human acute leukemia (AL) cells and their predictive value for chemotherapeutic efficacy. METHODS: MDM2 and p53 gene proteins expression was assayed by immunohistochemical staining. RESULTS: 1. The expression rates of MDM2 and p53 gene proteins were 71.7% and 21.7% respectively in 46 AL patients. The rates were slightly higher in relapse/refractory AL than in previously untreated AL; there was no difference among AL subtypes. 2. MDM2+ and p53- accounted for 67.4%, while the uniform expression of MDM2 and p53 15.2% (P < 0.01). 3. The marrow complete remission (CR) rate (69.2%) of MDM2- patients was higher than that (33.3%) of MDM2+ patients (P < 0.05). 4. Two of 4 patients with MDM2+++ gained CR and then MDM2 turned negative. CONCLUSION: MDM2 gene protein was negatively related with p53 gene protein in AL cells. Different expression patterns of the two gene proteins could influence the therapeutic efficacy, and combined detection of the two may be used as a prognostic parameter for AL patients.

Adolescent↗

PubMed1998

[The morphological and functional changes of pulmonary intravascular macrophages induced by LPS].

OBJECTIVE: To study the roles of pulmonary intravascular macrophages (PIMs) on infective acute lung injury (ALI). METHOD: Porcine pulmonary blood vessels were flushed by modified Morton's method, PIMs were isolated by adhesion method and identified with the features at both light and electron microscopic level. The activity of IL-1 beta, and content of IL-6, IL-8 and TNF alpha in the culture supernatants were measured by thymocyte proliferation or ELISA, respectively. RESULT: Enlarged and increased pseudopods, and increased amount of lysosomes and phagosomes were found in the PIM stimulated with lipopolysaccharide (LPS, 10 micrograms/ml); the releases of TNF alpha, IL-1 beta, IL-6 and IL-8 were increased significantly as compared with the level of pre-stimulation of LPS (P < 0.01), and reaching their peaks at 1 h, 2 h, 4 h and 6 h after LPS stimulation, respectively. CONCLUSION: The isolation of porcine PIMs can be completed with modified Morton's method; the phagocytosis and secretion of PIMs are more active after LPS stimulation. In the pathogenesis of ALI, TNF alpha and IL-1 beta may play an important role at its early stage; however, the IL-6 and IL-8 may be associated with the pathophysiological changes at the later stage of ALI.

Animals↗

PubMed1998

[Multilocular cyst of kidney].

OBJECTIVE: Multilocular cyst of kidney is a rare, unique and characteristic renal lesion with benign biologic behavior. We report a case of the disease. METHODS: It was confirmed by operation and pathology. RESULTS: It is difficult to distinguish radiologically multilocular cyst of kidney from cystic renal carcinoma and wilms tumor. CONCLUSION: Surgical intervention is the only effective treatment and method to differentiate multilocular cyst of kidney from a malignant lesion of the kidney.

Adult↗

PubMed1998

[Determination algorithms in oscillometric blood pressure measurement].

This paper discusses the method of oscillometric blood pressure determination. A new criterion for systolic pressure and diastolic pressure determination has been proposed, which is based on ratio changes of the oscillation amplitude under a wide normalized range. Primary experiment results show the new criterion is effective.

Algorithms↗

PubMed1997

Analysis of the Morgan-Elson chromogens by high-performance liquid chromatography.

The Morgan-Elson method for quantitative N-acetylhexosamine analysis is a two-step procedure comprising alkali treatment of the sugar and subsequent condensation of the resulting chromogens with p-dimethylaminobenzaldehyde (Ehrlich's reagent) to yield a colored product. In the present investigation, the products formed in the first step of the procedure were analyzed by high-performance liquid chromatography (HPLC) on a reversed-phase (C18) column, which was eluted with a water-methanol gradient; the absorbance of the effluent was monitored at 229 nm. The profile generated from alkali-treated N-acetylglucosamine exhibited two major peaks, in a ratio of approximately 2.5:1, which accounted for 94% of the total peak area. A third peak, accounting for 3% of the peak area, was eluted in an intermediate position, and several smaller peaks were also observed. The three predominant components, isolated by preparative HPLC, all gave a purple color on addition of Ehrlich's reagent, indicating that they were Morgan-Elson chromogens. The HPLC profile of alkali-treated N-acetylmannosamine was identical to that of the products generated from N-actylglucosamine, as was expected because of the elimination of the asymmetry at C-2 during formation of the chromogens. N-Acetylgalactosamine yielded two major peaks, which were eluted in the same positions as the two major products formed from N-acetylglucosamine, but the intermediate peak seen in the N-acetylglucosamine pattern was absent. The HPLC procedure allowed detection of as little as approximately 25 ng of N-acetylglucosamine and may therefore be of value as an alternative to the complete Morgan-Elson procedure when only small amounts of sample are available for quantitative analysis.

Acetylgalactosamine↗

PubMed1997

The involvement of sigma receptors in the choice reaction performance deficits induced by phencyclidine.

The present study investigated the effects of phencyclidine (PCP) on choice reaction in a 3-choice serial reaction time task for studying attentional function. PCP (3.2 mg/kg) significantly delayed choice reaction time and reduced choice accuracy. A novel sigma receptor antagonist N,N-dipropyl-2-[4-methoxy-3-(2-phenylethoxy)phenyl]-ethylamine monohydrochloride (NE-100) at 3.2 mg/kg, and 1-(cyclopropylmethyl)-4-[2'-4"-fluorophenyl)-2'-oxoethyl]piperidin e HBr (DuP734) at 1.0-3.2 mg/kg, but not 4-[2'-(4"-cyanophenyl)-2'-oxoethyl]-1-(cyclopropylmethyl)pip eridine (XJ448), antagonized both the delayed choice reaction time and the decreased choice accuracy elicited by PCP administration. The antagonism induced by NE-100 was blocked by the sigma receptor agonist (+)-N-allylnormetazocine HCl [(+)-SKF10,047]. These findings indicated that PCP (3.2 mg/kg) significantly induced attention deficit in a 3-choice serial reaction time task, and that this process may be mediated by sigma receptors.

Animals↗

PubMed1997

On the SAR and field inhomogeneity of birdcage coils loaded with the human head.

Birdcage coils are widely used as a radiofrequency (RF) resonator in magnetic resonance imaging (MRI) because of their capability to produce a highly homogeneous B1 field over a large volume within the coil. When they are employed for high-frequency MRI, the interaction between the electromagnetic field and the object to be imaged deteriorates the B1-field homogeneity and increases the specific absorption rate (SAR) in the object. To investigate this problem, a finite-element method (FEM) is developed to analyze the SAR and the B1 field in a two-dimensional (2D) model of a birdcage coil loaded with a 2D model of a human head. The electric field, magnetic field, and SAR distributions are shown, and a comprehensive study is carried out for both linear and quadrature birdcage coils at 64, 128, 171, and 256 MHz. It is shown that to generate the same value of the B1 field, the SAR is increased significantly with the frequency, and for the same imaging method the SAR produced by a quadrature coil is significantly lower than that of a linear coil. It is also shown that the B1-field inhomogeneity is increased significantly with the frequency.

Electromagnetic Fields↗