[Diagnosis of patent ductus arteriosus by 2-d echocardiography].
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Biomedical subjects
Publications and source records attributed to S Chen.
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Cells of the mouse cell line 3T3-F442A can be induced by various hormones to differentiate into adipocytes, whereas cells of 3T3-C2, a subclone of 3T3, cannot. However, transfection of DNA from uninduced 3T3-F422A cells into 3T3-C2 cells permits recovery of 3T3-C2 transfectants that differentiate into adipocytes in the presence of insulin. DNA isolated from human fat tissue, when transfected into 3T3-C2 mouse cells, also gives rise to mouse transfectants that are induced to differentiate into adipocytes by the addition of insulin. Apparently, transfection of a trans-regulatory gene (or genes) from 3T3-F442A or human fat cells into 3T3-C2 cells is sufficient to commit 3T3-C2 cells to adipocyte differentiation.
Injections of propidium iodide (PI) into the lateral cerebral ventricle of the rat resulted in a bilateral labeling in the septohippocampal nuclei, substantia nigra (SN), ventral tegmental area (VTA), retrorubral nuclei (rr), dorsal and median raphe nuclei, regions within and dorsal to the medial lemniscus of the caudal midbrain, and Purkinje cells in the cerebellum. No labeled neurons were seen in other areas of the brain. The data suggest that PI appears to exhibit selective labeling, and the mechanism underlying the selective labeling is discussed. Combined with Faglu histofluorescence, it was found that all PI-labeled cells in SN-VTA-rr were catecholamine (CA) neurons. After a transection of the medial forebrain bundle immediately before the PI injection, an accumulation of PI was only seen in the distal segments of severed nigrostriatal CA fibers. This provides a strong evidence that PI labeling of CA cells in SN-VTA-rr is due to axonal uptake and retrograde transport.
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Development dysgenesis of Purkinje cells or granule cells was analyzed for the reciprocal effect of reduced number of each cell type on the other. A single pre- or postnatal injection of methylazoxymethanol acetate (MAM) in the rat reduces either the number of Purkinje cells or the number of granule cells when administered at the time of their respective genesis. The total number of these two types of neurons was obtained from cell density values of each layer and the total volume of the granular layer and the area of the Purkinje cell layer. The results show that Purkinje cells (targets) strictly determine the maximum number of granule cells (afferent neurons) following deficits in the number of Purkinje cells produced by prenatal MAM administration. Deficits in Purkinje cells were accompanied by a proportionally smaller number of granule cells so that the ratio remained constant. On the other hand, the reduction in the number of granule cells of the postnatal MAM model did not affect the number of Purkinje cells. These results indicate that the maximum number of these afferent neurons is constrained unidirectionally through a property defined by the number of their target neurons which develop earlier. Furthermore the number of afferent cells had no effect on the number of target cells.
A monoclonal antibody, RS1-114, was raised against the human adenocarcinoma of the lung cell line A549. By studying the reactivity of RS1-114 with A549 cells following chemical and enzymatic treatments, it was shown that the epitope is a galactose-containing carbohydrate, which is devoid of sialic acid. Hemagglutination of desialylated RBCs, enzyme-linked immunosorbent assay studies with glycoprotein antigens before and after desialylation, and competition studies using peanut agglutinin indicate that monoclonal antibody RS1-114 recognizes the Thomsen-Friedenreich antigen, a cryptic determinant on human erythrocytes which can be exposed by neuraminidase treatment. It is expressed in an unhidden form on a large percentage of carcinomas and is therefore an important human tumor marker. RS1-114 is reactive with cryptic determinants of the Thomsen-Friedenreich antigen on white blood cells as well as red blood cells, and it reacts with unhidden determinants on human tumor cell lines. The number of binding sites on carcinoma cells is further increased by neuraminidase treatment. By immunohistochemical staining, it was shown that 75% of the human tumors tested are reactive with RS1-114. These include tumors of the breast, colon, lung, kidney, ovary, and rectum.
Eight phospholipidic classes from bovine brain cortex and soybean were prepared and purified by preparative high-performance and liquid chromatography, and their molecular species were identified by negative ion fast atom bombardment mass spectrometry using the 'surface precipitation' method. Its main advantages are: (i) clear-cut and abundant diagnostic ions for structural elucidation of the species; (ii) fragments characteristic of the main fatty acids, the polar head-group and the molecule frame in phospholipids; (iii) less background caused by the liquid matrix.
This article reviews 573 cases of acute pyrethroid poisoning reported in the Chinese medical literatures during 1983-1988. There were 325 cases of acute deltamethrin poisoning (occupational 158, accidental 167), 196 patients of acute fenvalerate poisoning (occupational 63, accidental 133, including 2 cases of ingestive fenvalerate-organophosphate mixture poisoning), 45 cases of acute cypermethrin poisoning (occupational 6, accidental 39) and 7 cases of other pyrethroid poisoning (occupational 2, accidental 5). The clinical manifestations have been reviewed and analysed. The diagnosis, gradation and differential diagnosis of occupational acute pyrethroid poisoning have been discussed.
The immunosuppressive activity of azathioprine (AZA) is unpredictable and depends on the formation of intracellular thiopurine ribonucleotides. However, the quantification of these active thiopurines presents difficult analytical problems. It has recently been postulated that plasma concentrations of 6-thiouric acid (6-TU) and 6-mercapto-purine (6-MP), metabolites of AZA, may provide more readily measurable indices of the pharmacologic activity of AZA. In order to evaluate the utility of 6-TU and 6-MP plasma concentrations in monitoring AZA therapy, we studied their pharmacokinetics in 6 renal transplant patients, and their in vitro immunosuppressive potency in a mixed lymphocyte proliferation assay. A peak plasma 6-TU concentration of 710.7 ng/ml was observed at 3.8 h after oral dosing. Good correlation was observed between the elimination t1/2 of 6-TU and serum creatinine, and between AUC over 24 h and serum creatinine. However, we did not observe a second peak in plasma 6-TU concentration that could be attributed to the degradation of active AZA metabolites. 6-MP plasma concentrations in the patients were low (mean peak concentration 36.0 ng/ml) and rapidly disappeared within 8 h. In vitro immunosuppressive activity could not be demonstrated for 6-TU over a concentration range of 1.25 ng/ml to 0.25 mg/ml. We conclude that 6-TU is pharmacologically inert and is primarily eliminated by the kidneys. Our findings currently do not support the use of plasma concentrations of 6-TU or 6-MP to monitor AZA therapy. In order to optimize AZA therapy, analytical techniques that are technically feasible and that can directly quantify the active intracellular thiopurines are being explored.
Surface electrocardiogram (ECG), sinus node electrogram (SNE), His bundle electrogram (HBE), and intracardiac electrogram of high right atrium (HRA) were recorded synchronously in 11 of the 14 consecutive cases with complete atrioventricular block (CAVB). SNE showed the presence of a pre-P wave that preceded in all the P wave on the ECG and had no relationship to QRS complex or TU repolarization. This confirmed that the pre-P wave arose from the sinus node. Direct sinoatrial conduction time (SACTd) in nine cases was within the normal range (less than 120 msec). SACTd in another two cases was prolonged to 185 msec and 210 msec, respectively, suggesting their abnormal sinus node conduction.
Pretreatment regimens directed at reducing the immunogenicity of pancreatic islets have emphasized the elimination or alteration of the major histocompatibility complex (MHC) class II-positive dendritic cells within the islet. Unfortunately, the efficacy of such pretreatment regimens has been extremely variable and the relative contribution of the dendritic cells to the overall immunogenicity of pancreatic islets has remained ambiguous. Recent evidence has suggested that the MHC class I antigen present on the endocrine cells within the islets may play an important role in the alloimmune response. This study utilized the in vitro mixed lymphocyte-islet co-culture system to determine if pretreatment of whole islets with an anti-MHC class I monoclonal antibody specific to the donor strain would block the generation of cytotoxic T lymphocytes (CTL) in the in vitro mixed lymphocyte-islet coculture. Pretreatment of B10.BR (H-2k) and DBA/2J (H-2d) islets with an allospecific anti-MHC class I monoclonal antibody blocked the generation of allospecific CTL when the pretreated islets were placed into coculture with C57Bl/6 (H-2b) splenocytes. If such a pretreatment regimen is similarly effective in vivo, it could potentially be used as an antirejection strategy in pancreatic islet allotransplantation.
We studied the differentiation of acute promyelocytic leukemia (APL) cells in 14 patients with APL. After the induction by retinoic acid (RA) the mature cells rose to 60 +/- 11.8% compared to 0.7 +/- 1% of the control, while the promyelocytes declined to 8.7 +/- 6.4% (93.3 +/- 5.6% in the control group). Protein kinase C (PKC) activity was significantly increased to 149.3 +/- 156.2 pmol/mg per min compared to 47 +/- 40.9 of the control (p less than 0.01). In HL-60 cells, the activity of PKC increased also from 52.3 +/- 35 to 129.2 +/- 64.6 pmol/mg per min (n = 10, p less than 0.01) after the induction of differentiation with RA. If the leukemia cells were pretreated with a kind of PKC inhibitor such as trifluoperazine, the increase of PKC activity was inhibited, and the rate of nitroblue tetrazolium reduction decreased from 89.9 +/- 7.7% to 62 +/- 25% (n = 6, p less than 0.01) and the mature cells reduced from 63.1 +/- 11.7% to 19.7 +/- 12.2% (p less than 0.01). We presumed that the activity of PKC is closely related to the differentiation of human promyelocytic leukemia cells induced by all-trans-retinoic acid.
We have previously reported that purified hepatocytes stimulate significant in vitro allospecific cytotoxicity when cocultured with naive responder splenocytes in the mixed lymphocyte hepatocyte culture (MLHC). In this report we examined the expression of MHC antigens on the surface of hepatocytes, the phenotypic lymphocyte subset(s) that respond(s) to allogeneic hepatocytes, and the phenotype of allospecific cytolytic effectors generated in MLHC. Hepatocytes expressed MHC class I but not MHC class II antigens by immunofluorescent microscopy and fluorescence activated cell sorting. The lack of MHC class II on the surface of hepatocytes was also indirectly supported by the inability of hepatocytes to stimulate proliferation of a class II-directed allospecific helper T cell clone. The generation of allospecific cytotoxicity in MLHC required the participation of L3T4+, Ly2- T cells and L3T4-, Ly2+ T cells in the naive responder splenocyte population since depletion of these subsets with mAb and complement abrogated the development of allo-CTLs. Furthermore, adherent accessory cells in the naive responder splenocyte population appeared to play a role in the generation of allospecific cytotoxicity in MLHC since depletion of this population by plastic adherence and passage through a Sephadex G10 column resulted in significantly reduced allospecific cytotoxicity. Depletion of day 5 allosensitized cells of Ly2+ but not L3T4+ T cells by mAb and complement eliminated allospecific cytotoxicity--indicating that cytolytic effectors generated in MLHC appear to be L3T4-, Ly2+ T cells.
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A full-length human placental aromatase cDNA clone, Aro 2, was isolated upon screening a human placental cDNA library with an aromatase cDNA probe and an oligonucleotide probe whose sequence was derived from a human aromatase genomic clone. Nucleotide sequence microheterogeneity was found in the 3'-untranslated region among Aro 2 and in two previously described human aromatase cDNA clones. Both the minor sequence differences and the expression of a single protein species in placental tissue suggest the presence of different alleles for aromatase. Northern blot analyses using one cDNA and two oligonucleotide probes are consistent with the two mRNA messages of 2.9 and 2.5 kilobases arising in human placenta as a consequence of differential processing. Several yeast expression plasmids containing the aromatase cDNA we cloned were constructed. The enzyme was expressed in Saccharomyces cerevisiae. The expressed activity was inhibited by the known aromatase inhibitor, 4-hydroxyandrostenedione. A level of 2 micrograms aromatase/mg partially purified yeast microsomes was estimated by analyses of carbon monoxide difference spectra on microsomal fractions from yeast carrying plasmid pHARK/VGAL. Using [1 beta, 2 beta-3H]androst-4-ene-3,17-dione as the substrate, an apparent Michaels-Menken constant (Km) of 34 nM and a maximum velocity (Vmax) of 23 pmol [3H]water formed per min/mg protein were obtained for the yeast synthesized aromatase by transformation with plasmid pHARK/VGAL. The kinetic results are similar to those determined for human placental aromatase, and suggest that the yeast synthesized aromatase will be useful for further structure-function studies.