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[Effect of monoamine neurotransmitters in the hypothalamus in a cortisol-induced rat model of yang-deficiency].

Eight monoamine neurotransmitters in hypothalamus of normal and cortisol-induced Yang-deficiency rats were measured with HPLC-ECD. These neurotransmitters included: (1) Noradrenaline (NA); (2) Adrenaline (A); (3) 3-methoxy-4-hydroxyphenyl glycol (MHPG); (4) 3,4-dihydroxyphenylacetic acid (DOPAC); (5) Dopamine (DA); (6) 5-hydroxyindole-3-acetic acid (5-HIAA); (7) Homovanillic acid (HVA); and (8) Serotonin (5-HT). With the mobile phase of 10% methanol-0.15 M chloroacetic acid-sodium hydroxide buffer, the neurotransmitters might be well separated, the sensitivity was similar to that of gas chromatography-mass spectrogram with the lower limit of 0.1 ng, this assay was simple and might be accomplished with half an hour. The Yang-strengthening Chinese herbs Radix Aconiti Praeparata, Cortex, Cinamomi, Epimedium brevicorum and Herba Cistanchis were used in the study and the following were founded: (1) In the hypothalamus of cortisol-induced Yang-deficiency rat NA decreased and a increased when compared with that of the normal controls (both P less than 0.05). (2) After 4 weeks of administration of Yang-strengthening herbs, the NA and A of the Yang-deficiency rats returned to nearly normal and it suggested these herbs have the effect of "supporting the right and recovering the source"; DA markedly increased both in control rats and Yang-deficiency rats (P less than 0.05, P less than 0.001 respectively); DOPAC obviously decreased both in control rats and Yang-deficiency rats (P less than 0.001); 5-HT and 5-HIAA increased in the control rats (both P less than 0.001) and HVA significantly decreased in Yang-deficiency rats. (3) The ratio of DA/DOPAC and 5-HT/5-HIAA markedly increased both in the control and Yang-deficiency rats after using Yang-strengthening herbs (P less than 0.01-0.001) and it suggested Yang-strengthening herbs could inhibit the activity of monoamine oxidase in hypothalamus.

3,4-Dihydroxyphenylacetic Acid↗

[Esterase activity of neutrophils and monocytes in patients with stenocardia and myocardial infarction].

Neutrophils and monocytes of coronary patients and healthy subjects were investigated for activity of alpha-naphthylacetate esterase, naphthol--AS--D+chloroacetate esterase, alpha-naphthylbutyrate esterase. This activity varied from healthy to coronary subjects, among anginal and MI sufferers, etc. The fact made the authors to suggest comprehensive evaluation of monocyte and neutrophil esterase activity as a diagnostic and prognostic indicator.

Adult↗

Effects of culture matrix on differentiation of murine mast cells.

The effects of culture matrix (agar, collagen, and methylcellulose) on differentiation of mast cells were investigated. Because berberine sulfate-positive colonies in agar were composed of macrophages but not of mast cells, the naphthol AS-D chloroacetate esterase reaction was used to identify mast-cell colonies. When bone marrow cells of WBB6F1 mice were cultured with conditioned media containing interleukin 3 (IL-3), numbers of mast-cell colonies were greater in collagen cultures than in agar and methylcellulose cultures. Electron microscopic examination revealed that mast cells developing in collagen and agar cultures were more mature than those developing in methylcellulose cultures. However, when bone marrow-derived cultured mast cells or purified peritoneal mast cells were plated, the efficiency of colony formation and size of colonies were comparable among agar, collagen, and methylcellulose cultures. Therefore, all three matrices tested had similar effects on the proliferation of mast cells. Collagen appeared to be suitable for differentiation of bone-marrow precursors and their maturation. Agar appeared to be suitable only for maturation.

Agar↗

[Preparations of preoral sustained-release preparations with a biological polymer base. 5 (correct title supplied from Table of Contents)].

Triethylamine and chloroacetic acid were used for the preparation of polyglycolic acid. The polyester was characterized by elementary analysis, IR-spectroscopy, X-ray-test and the determination of the molecular mass. With regard to the pharmaceutical and technological characterization the determination of the grain size, moisture content, flowability and the parameters for the direct compression were carried out. Polyglycolic acid was compressed together with caffeine as model drug. These tablets distinguished by a great strength, but in water a rapid decay took place. A greater stability in artificial gastrointestinal fluids was obtained, if the tablets were plastified. Release investigations showed then the desired retard effect.

Acrylic Resins↗

[Experimental study of a new calcification reducing monocusp patch].

The glutaraldehyde (GA)-treated porcine pericardium was modified chemically with epoxychlopropane (EC). Monocusp patches were made and implanted in the right ventricular outflow tract of dogs (n = 10). The GA or chloroacetic treated porcine pericardium monocusp patches was used as the control groups. At 6 months after implantation, the calcium of the samples were detected qualitatively (by von Kossa staining technique and with light microscopic observation), quantitatively (by a tomic absorption) and ultrastructurally. The stability and mechanical properties of tissue after modification with EC were assessed. The results of this study prove that chemically modified porcine pericardium with EC can reduce the calcification of tissue, and can preserve its structural intactness, good tissue stability, good tensile strength and function.

Animals↗

Correlation between morphological and cytochemical heterogeneity of acute promyelocytic leukemia (APL) and its association with myelodysplasia.

Sixty one cases of acute promyelocytic leukemia (M3) were diagnosed between Jan., 1985 and Dec., 1988. Morphologically 46 cases (75.4%) were characterised as typical M3 and 15 cases (24.6%) as M3 variant. Typical M3 cases had higher number of hypergranular promyelocytes and Auer rods (P less than 0.001). The cytochemical stains of myeloperoxidase, Sudan Block and Black and chloroacetate esterase were strongly positive in M3 typical and mild to moderately positive in M3 variant cases. Alpha-naphthyl acetate esterase positivity with fluoride inhibition was seen only in M3 variant cases (80%). The clinical and haematological parameters including marrow blast count were not significantly different in the two groups. This study has shown that M3 variant cases more frequently express heteregenous cytochemical patterns and myelodysplastic changes.

Adolescent↗

Phenotypically distinct target cells for murine sarcoma virus and murine leukemia virus marrow transformation in vitro.

An in vitro hematopoietic microenvironment was established from explained fragments of bone marrow from adult noninbred NIH Swiss mice with the use of corticosteroid-reconstituted horse serum. Infection with Kirsten murine sarcoma virus (Ki-MuSV) with either a Rauscher murine leukemia virus (R-MuLV) or Balb:virus-1 helper virus coat reduced proliferation of granulocytic and pluripotent hematopoietic stem cells and produced neoplastic transformation of both macrophages and preadipocytes in the adherent cell population within a 4-week period. Ki-MuSV-transformed, virus-releasing macrophages formed clusters of 4-49 cells in 0.8% methylcellulose-containing medium in the absence of added colony-stimulating factor (CSF), synthesized lysozyme, ASD-chloroacetate substrate-specific esterase-M, and CSF, and produced tumors following inoculation iv into adult NIH Swiss mice or ip into newborn NIH Swiss mice. In cultures infected with helper leukemia viruses R-MuLV or Balb:virus-1, gradual transformation of a distinct cell phenotype was observed over a 9-week period with generation of increasing numbers of atypical myeloblasts and promyelocytes which showed dyssynchronous nuclear-cytoplasmic maturation, basophilic granulation, cytoplasmic vacuolation, and formation of incompletely maturing CSF-dependent granulocyte-macrophage colonies in vitro and small spleen colonies in vivo. These data demonstrated that rapid biologic expression of the murine sarcoma virus genome in specific adherent "stromal" marrow cells prevents detection of a more subtle helper-virus-induced dysmyelopoiesis in a distinct nonadherent cell population.

Animals↗

A new hematopoietic cell line, KMT-2, having human interleukin-3 receptors.

A novel human cell line, KMT-2, from umbilical cord blood cells was established based on the selection of cultures in the presence of recombinant human interleukin-3 (IL-3) and the sorting of cells with anti-My 10 antibody. Morphologic and cytochemical studies (peroxidase negative, Sudan-black negative, chloroacetate esterase negative, PAS positive, nonspecific esterase positive) and phenotyping (HLA-DR, My7 = CD13, My9 = CD33, My10 = CD34, MCS-2, LeuM1 positive, glycophorin A negative, and P2 negative) suggest that the KMT-2 cells are myelomonocytic cells, probably of immature progenitor origin. Besides IL-3, granulocyte-macrophage colony-stimulating factor supported the growth of the KMT-2 cells, but IL-1 alpha, IL-2, IL-4, IL-5, and erythropoietin did not. IL-6 showed only slight activity. Binding studies with 125I-labeled recombinant human (rh) IL-3 indicated that IL-3 bound to a single class of high affinity receptors (approximately 4,000 receptors/cell) on KMT-2 cells with a kd of approximately 200 pmol/L. The chemical cross-linking assay demonstrated that radiolabeled hIL-3 bound three molecules with molecular masses of 170, 130, and 70 Kd. Present data suggest that the newly established human cell line will be a valuable tool for the biologic assay of hIL-3, and a model for biochemical studies of IL-3 receptors.

Blood Cells↗

[Quantitative studies of mast cell proliferation at the wound edge --rate of DNA synthesis in intravital and postmortem biopsy].

The number of DNA synthesizing mast cells in experimental induced wounds was calculated using bromodeoxyuridine as a DNA marker and naphthol AS-D chloroacetate esterase as a mast cell marker. After different survival times, specimens of the wound edge were taken before death and 24 hs after death. At a survival time of 48 hs in vital biopsy the number of DNA synthesizing mast cells peaked, at 72 hs and fell after a period of 8 days. Even though an identical kinetic course was demonstrable in cases of postmortal biopsies the percentage at each survival period was significantly lower than in vital biopsies.

Animals↗

[Synthesis of oligosaccharides with group specificity for blood types A, B and H (Type 3)].

Chemical synthesis of A, B, and H (type 3) human blood group determinant oligosaccharides (as R-glycosides, R = OCH2CH2CH2NHCOCF3) and their polymeric derivatives are reported. 4,6; 4',6'-Di-O-benzylidene derivative of Gal beta 1----3GalNAc alpha 1----R was chloroacetylated selectively at 3'-OH, the chloroacetate was alpha-fucosylated and dechloroacetylated to give protected H (type 3) trisaccharide bearing free 3'-OH. alpha-Glycosylation of the trisaccharide with 2-azido-3,4,6-tri-O-benzyl-beta-D-galactopyranosyl chloride and 2,3,4,6-tetra-O-benzyl-alpha-D-galactopyranosyl bromide gave rise to protected A and B tetrasaccharides, respectively. Deprotected R-glycosides were converted to OCH2CH2CH2NH2 derivatives. Their reaction with poly(4-nitrophenylacrylate) affords polyacrylamide-coupled conjugates with A, B, and H (type 3) specificity.

ABO Blood-Group System↗

[Acute myelomonocytic leukemia with mastocytosis in bone marrow].

A 22-year-old female was admitted to our hospital because of general fatigue. The lymph nodes, liver and spleen were not palpable. She was without cutaneous lesions. Haematological examinations revealed leukocytes 3,200/microliters with 44% blasts of myelomonocytic origin, and platelets 15,000/microliters. Bone marrow smears were hypercellular marrow with 51% blasts of myelomonocytic origin and focal involvement of mast cells. Serum histamine and vitamin B12 level was high. Mast cells were round with rounded or segmented nuclei. The nucleoli were inconspicuous and the cytoplasm contained a number of metachromatic granules. Cytochemically, mast cells stained positive for alpha-naphthol-AS.D-chloroacetate esterase and acid phosphatase, and negative for peroxidase, Sudan black B and alpha-naphthyl butylate esterase. In toluidine blue staining, mast cells had stained similarly with pH values from 2.5 to 6.5. She was diagnosed as acute myelomonocytic leukemia with benign mastocytosis, and treated with BH.AC-DNP. A complete remission was obtained, but mast cells in the marrow did not decrease. Relationship between leukemia and mastocytosis was not known, but it was suggested that mast cells responded to the proliferation of the leukemic cells.

Adult↗

[Non-specific esterase positive neutrophils in a case of refractory anemia with excess of blasts].

A 61-year-old woman was admitted with complaint of fever. The peripheral blood showed pancytopenia and bone marrow aspirate showed dysplasia in trilineage blood cells with increased blasts (18.2%). Bone marrow chromosome study revealed a karyotype of 46XX, -6, 3q-, +mar in 19 cells of 20 analyzed. She was diagnosed as refractory anemia with excess of blasts (RAEB). 95% of neutrophils in the bone marrow and 84% of that in the peripheral blood were stained with non-specific esterase using alpha-naphthyl acetate as substrate. On the other hand, the positivity of neutrophils for peroxidase. Sudan black B or chloroacetate esterase was markedly decreased. The phagocytotic activity of neutrophils was increased in comparison with normal control cells. Surface marker analysis of peripheral blood myeloid cells revealed increased expression of monocyte specific markers. These findings suggested that the patient's neutrophils, which were surely neutrophils in morphology, shared also monocyte-specific characters. After treatment with low dose Ara-C, pancytopenia was recovered and blasts in the bone marrow were reduced. Also was decreased non-specific esterase positive neutrophils, indicating that the neutrophils were derived from abnormal myeloid clone.

Anemia, Refractory, with Excess of Blasts↗

Cytochemistry of acute promyelocytic leukemia (M3): leukemic promyelocytes exhibit heterogeneous patterns in cellular differentiation.

Cytochemical investigation of leukemic promyelocytes from 25 cases of acute promyelocytic leukemia (M3) disclosed two major cellular differentiation categories: (1) the pure neutrophilic (N) type (16 cases) with strong myeloperoxidase (MPO) and naphthol-ASD chloroacetate esterase (Es-chl), but lacking the monocytic enzyme NaF-sensitive alpha-naphthyl butyrate esterase (Es-b), and (2) the mixed neutrophilic/monocytoid (N/M) type (seven cases) with strong Es-b as well as strong MPO, all cases exhibiting Es-dual (Es-b + Es-chl) positive cells. Two more cases with unusual phenotypes were noted: one with intense lysozyme activity but without Es-b and the other with toluidine blue-methachromasia and negative MPO. Promyelocytes from the control group, consisting of nine cases of t(8;21) M2 AML and ten cases with normal bone marrow, lacked such cytochemical heterogeneity. HL-60, an M3 cell line that can be induced to differentiate toward monocytic lineage in vitro, was almost negative for Es-b in the uninduced condition. Cytogenetically, eight cases of N type and five of N/M type had the t(15;17) abnormality. Thus at least two differentiation patterns were observed in M3 leukemia with fidelity (N type) and infidelity (N/M type) for normal granulocytic differentiation. In this series, there was no statistically significant difference in clinical features (remission rate and survival) between the two types. Our study suggests that the development of M3 leukemia is not exclusively restricted to the neutrophilic pathway, but more heterogeneously related to myelomonocytic differentiation.

Adolescent↗

[A light and electron microscopic study of acute myelomonocytic leukemia cells in comparison with normal monocytes and acute monocytic leukemia cells].

Leukemic cells from 15 patients with acute myelomonocytic leukemia (M4) were investigated by light and electron microscopy. As controls, the blood cells from 2 normal volunteers and 3 acute monocytic leukemia (M5b) were examined. To identify monocytic cells, a double esterase (DE) staining combining with alpha-naphthyl butyrate esterase (ANBE) and naphthol-ASD chloroacetate esterase (N-ASD-CAE) stainings was used. According to DE staining, M4 leukemic cells were classified into ANBE positive cells, N-ASD-CAE positive cells and DE-positive cells coated vesides. Ultrastructurally, M4 leukemic cells possessed indented nuclear membrane, clusters of myeloperoxidase-positive rod-like granules, coated vesicles, abundant smooth endoplasmic vesicles and invaginated cell membrane similarly to M5b leukemic cells and normal monocytes. These features were observed not only in the ANBE positive cells but also in the DE-positive cells. The N-ASD-CAE positive cells in M4 ultrastructurally showed atypical features of polymorphonuclear leukocytes and possessed monocytic characteristics to some extents. M4 leukemic cells occasionally showed some peculiar structures such as the reticulated lamellae complex, paired cisternae and annulate lamellae. The cells showing erythrophagocytosis were either observed. Combining the results of DE staining and electron microscopy, it was suggested that M4 is closely associated with the malignant transformation of a monocyte/granulocyte precursor cell which has an ability to differentiate to both monocyte and granulocyte.

Adult↗

[Differential diagnosis in the preliminary evaluation of esterase responses of tongue coating cells].

The tongue coating cells were stained by Alpha-naphthol acetate esterase and naphthol AS-D chloroacetate esterase in 100 patients with different symptom-complex [five groups: dampheat, damp-heat due to blood stasis, Yin deficiency, Yang deficiency and Qi-blood deficiency] in view of TCM, and in 116 healthy persons. The responses of the epithelial cells of the tongue and the leukocytes to the two esterase stainings were observed. The results were: the number of nonspecific esterase was much higher in the patients, especially in the patients with Yin or Qi-blood deficiency, than that in the healthy persons (P less than 0.001); responses to ASDCE suggested that the leukocytes of the patients were different from those of the healthy persons in classification, more granulocytes in the healthy persons, and increased proportion of monocytes and lymphocytes in the patients, the most increased proportion of monocytes in the patients with Qi-blood deficiency and the most increased proportion of lymphocytes in the patients with Yang deficiency. The mechanisms of the different responses to the two esterase stainings in the patients and healthy persons, and the relationship between these mechanisms and the symptom-complexes by TCM were discussed.

Adolescent↗

Prognostic significance of myeloperoxidase positivity of blast cells in acute myeloblastic leukemia without maturation (FAB: M1): an ECOG study.

The relationship between cell features by light microscopy and therapeutic outcome of 72 patients with acute myeloblastic leukemia (FAB M1) were investigated. The patients were divided into two groups according to myeloperoxidase-(MPO) positive percentage of blast cells, namely a low (less than 50%) and high (greater than 50%) MPO group. No remarkable morphological difference of blast cells between these two groups was observed. The 38 patients with low MPO showed a significantly lower complete response rate (CR) (52.6%) than the 34 patients with high MPO (85.3%) (p = 0.003) that included 10 CAE-positive patients and 2 ANAE-positive CAE-negative patients. The low MPO group included 5 CAE-positive patients with granulocyte dysplasia and 7 additional patients with alpha naphthyl acetate esterase (ANAE) positivity who were chloroacetate esterase (CAE) negative. Low positivity of blast cells may be due to a defective enzyme expression in the former but also may be a reflection of 'monocytic' aberrant expression in the latter. The low MPO group in M1 with this heterogeneous population is less likely to achieve CR with chemotherapy, while the high MPO group with a higher CR rate suggests a more pure entity within the myeloblastic leukemias referred to as M1. After further refinement by eliminating 24 cases that were 'esterase positive' (ANAE and/or CAE), the results were still the same, suggesting that the more immature blast populations were in the low MPO group.

Adult↗

[Chronic myelomonocytic leukemia (CMMoL) with systemic lymph node swelling and paroxysmal nocturnal hemoglobinuria (PNH)-like complication].

A 37-year-old male was diagnosed as having chronic myelomonocytic leukemia (CMMoL) with chief complaint of systemic lymph node swelling. On admission, his peripheral blood revealed mild anemia and mild thrombocytopenia with giant platelets, and monocytosis (1480/microliters). NAP score was low. Serum lysozyme increased. The bone marrow showed normal cellularity consisting of 4% myeloblasts and 14.4% promyelocytes, and a few myeloid cells were positive for double staining by alpha-naphthyl butyrate and naphthol ASD chloroacetate esterase. Biopsied specimens of the cervical lymph node showed infiltration of monocytoid cells, which were positive for lysozyme staining, into interfollicular tissue. As for chromosome variation, 21 large satellite was observed in all dividing cells from his bone marrow and peripheral blood. Furthermore, hemolytic anemia with hemoglobinuria developed during his course. Sugar water test was positive, but Ham test negative. Coombs test and Donath-Landsteiner reaction were negative. Abnormal hemoglobin, spherocyte and fragmentation were not found. Hemolysis disappeared about two months later. However, blastic crisis appeared and he died. We showed a case of CMMoL with 21 large satellite and paroxysmal nocturnal hemoglobinuria (PNH)-like complication. Satellite have usually been reported as asymptomatic, and thus this chromosome variant and CMMoL may have been coincidentally observed.

Adult↗

[Relapse of acute monocytic leukemia present with skin and tracheal involvement].

A 34 year-old female was admitted because of anemia and leukopenia. Her bone marrow contained abundant blastic cells, which were histochemically positive for peroxidase and alpha-naphthyl butyrate esterase, but negative for ASD chloroacetate esterase. She was diagnosed as acute monocytic leukemia (FAB, M5a). Complete remission was achieved after the administration of BHAC, daunorubicin, 6MP and prednisolone, and she was discharged after consolidation therapies. But shortly later, she noticed hoarseness and erythematous nodules on her breast and abdomen. Though the examinations of peripheral blood and bone marrow did not show any abnormality, hoarseness rapidly worsened and she complained of dyspnea. X-ray and CT scan demonstrated narrowing of the trachea under the cricoid cartilage, and trans-tracheal biopsy revealed leukemic involvement. In addition, erythematous skin lesion showed the infiltration of leukemic cells by biopsy. Although radiation and chemotherapy was initiated, she died of pneumonia. We tried to discuss the laryngo-tracheal and skin involvement of acute monocytic leukemia as early symptoms of relapse.

Adult↗