[Detection of Hepcidin in transfusion dependent myelodysplastic syndrome patients and its clinical significance].
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To explore the application value of detection of Hepcidin together with indicator of iron overload on clinical diagnosis and treatment of MDS with iron overload by measuring Hepcidin and iron load indices of transfusion dependent myelodysplastic syndrome (MDS) patients.Enzyme-linked immunosorbent assay (ELISA), radioimmunoassay and colorimetry were used to determine the Hepcidin, serum ferritin (SF) and serum iron (SI) levels of 106 serum samples from 68 cases of transfusion dependent MDS patients, 30 serum samples of MDS patients without transfusion and 60 serum samples of controls.For MDS group, Hepcidin level in blood transfusion < 9 U subgroup was significantly higher than that in control group \[(583 ± 50) µg/L vs (175 ± 35) µg/L\] and there was a strong positive correlation between Hepcidin levels and SF (r = 0.976), but no correlation between Hepcidin and SI (r = 0.284); Both Hepcidin and SF level in transfusion 9 ∼ 24 U subgroup was significantly higher than those in control group \[(665 ± 80) µg/L vs (175 ± 35) µg/L; (1445 ± 275) µg/L vs (112 ± 26)µg/L\]; whereas for SI level, there was no difference between transfusion 9 ∼ 24 U subgroup and the control group. Hepcidin did not correlate with SF or SI; For blood transfusion > 24 U group, all of Hepcidin, SF and SI levels were higher than those in control groups \[(703 ± 64) µg/L vs (175 ± 35) µg/L; (2587 ± 352) µg/L vs (112 ± 26)µg/L; (20 ± 4) µg/L vs (14 ± 4) µmol/L\], Hepcidin negatively correlated with SF and SI (r = -0.536; r = -0.456). Hepcidin levels of RARS patients were significantly lower than RAEB patients \[(260 ± 40) µg/L vs (442 ± 51) µg/L\], and there was no significant difference between RARS group and control group regardless of the number of blood transfusion.Both Hepcidin and SF levels in MDS patients regardless of transfusion dependent or not, or the number of blood transfused were higher than those of normal controls, the increase of Hepcidin can not synchronize with the increase of SF level due to the increased blood transfusion, when blood transfusion > 24 U, Hepcidin level showed a negative relationship with SF and SI, reflecting the decreased ability of Hepcidin to inhibit body iron absorption during the increase of blood transfusion, which finally would lead to iron overload. We can predict the occurrence of iron overload in transfusion dependent MDS patients by dynamic monitoring concentration of Hepcidin.Keywords:
Clinical Significance
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In 3 separate studies, each spanning 32–50 hr, serum and pituitary LH levels of male rats killed at 1½ to 6½ hr intervals were studied by radioimmunoassay. In 2 of these studies orchidectomized rats also were examined and in one of them (spanning 50 hr) ovariectomized rats were studied as well. In no case was evidence obtained for any rhythmic diurnal variation of either serum or pituitary LH levels. In the third study, on male rats only, serum and pituitary FSH were examined by 3 different radioimmunoassay systems. Serum corticosterone and pineal serotonin levels also were measured. The animals displayed pineal serotonin and serum corticosterone rhythms very similar to those other male rats have been reported to exhibit, but they showed no sign of any rhythmic variation in pituitary or serum FSH levels. These findings lead us to conclude that: 1) the rates of LH and FSH secretion remain fairly constant throughout the day and night in intact rats, even though such rats do display striking diurnal rhythms of pineal and adrenal activity; 2) LH secretion is also fairly stable in castrated rats of both sexes; 3) any rhythmic variations which may exist in the serum and pituitary hormone levels we studied are of such small amplitude that they were not detectable by the methods we used. (Endocrinology87: 798, 1970)
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Although insulin-like growth factor-binding protein-2 (IGFBP-2) is an abundant IGFBP in fetal and postnatal plasma, its regulation is not yet clearly understood. To address this question in sheep, we purified ovine IGFBP-2 and developed a homologous radioimmunoassay. We have studied its ontogenesis and measured serum concentrations of ovine IGFBP-2 after bovine growth hormone (bGH), ovine placental lactogen (oPL) and IGF-I treatment. Concentrations of IGFBP-2 were high at 125 days of gestation (550 +/- 15 micrograms/l) but fell after birth (P < 0.05) and plateaued after 1 year of age (340 +/- 20 micrograms/l). In lactating ewes, bGH treatment for 7 days significantly reduced (21%; P < 0.05) IGFBP-2 relative to the saline-treated group. Similarly, in neonatal lambs, bGH treatment from day 3 to day 23 of life reduced (P < 0.05) IGFBP-2 by 23% relative to the saline-treated group. oPL had no effect on serum levels of IGFBP-2 in the ewe or the neonatal lamb. In well-fed yearling lambs, treatment with IGF-I reduced IGFBP-2 values by 27% (P < 0.05) relative to control animals. In yearling lambs, reduced nutrition increased plasma IGFBP-2 (41%; P < 0.05). However this increase was abolished by IGF-I treatment. The changes in plasma levels of IGFBP-2 were positively related to changes in IGF-II while there was a negative relationship between circulating IGF-I and IGFBP-2 such that both IGF-I and IGF-II may play a role in the regulation of IGFBP-2 in serum.
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A sensitive homologous radioimmunoassay for mouse GH capable of measuring concentrations of GH as low as 1 ng/ml of mouse serum has been developed. This assay has been used to study GH secretion in C3H mice under several physiological conditions. Newborn mice possessed very high concentration of GH in serum which persisted for two weeks. Adult male mice had more GH in their pituitary (66.9 μg/mg) and serum (12.0 ng/ml) than comparable females (48.9 μg/mg; 3.4 ng/ml). Ovariectomy in female mice increased pituitary and serum concentrations of GH to male levels whereas stilbestrol administration decreased it. Orchiectomy or stilbestrol treatment of male mice resulted in decreased GH levels. During the estrous cycle, maximum levels of GH were attained during proestrus. Insulininduced hypoglycemia or treatment with L—dopa failed to induce rise in serum concentrations of GH in mice.(Endocrinology91: 784, 1972)
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SUMMARY A radioreceptor assay (RRA) for GH, using pregnant rabbit liver membranes, was used to evaluate the receptor‐hormone interaction of circulating (serum) GH in diabetics. Multiple morning blood samples were taken from 18 fasted insulin‐dependent diabetics on three occasions and assayed for GH by the RRA and radioimmunoassay (RIA). Samples from 12 normal subjects (GH secretion stimulated by exercise and hypoglycaemia) and 11 acromegalics were also studied. The RRA/RIA ratio was higher in samples from the acromegalics than in normal subjects (0.85 vs 0.65, P < 0.01) and only a few samples had RRA values greater than RIA. All the samples from 14 of the diabetics had lower RRA values than RIA, but 40% (39 of 96) of samples from the remaining four diabetics had RRA concentrations markedly in excess of the levels measured by RIA. Sephadex chromatography (G100) of serum samples revealed similar proportions of immunoreactive GH forms in the diabetics with high RRA values compared with the diabetics with low RRA values. These findings may indicate intermittent secretion of highly bioactive GH in some insulin‐dependent diabetics. Further studies on the biological properties of circulating GH are needed to clarify the role of GH secretion in the development of diabetic microvascular complications.
Sephadex
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Transferrin saturation
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In the present study the influence of dexamethasone treatment of rats on the basal values of thyrotrophin (TSH) and prolactin (PRL) and the response of both hormones to thyrotrophin releasing hormone (TRH) has been investigated. Male rats were given 100 μg of dexamethasone/rat for 8 days at the same time of day. Four hours after the last administration of dexamethasone 200 ng or 100 μg of TRH/rat was injected ip. Blood was collected 10 min later by decapitation. TSH and PRL were estimated by radioimmunoassay (RIA) using the NIAMD kits. The basal and TRH stimulated values of PRL in plasma were significantly lower in dexamethasone treated rats than in controls (P < 0.01). The basal TSH levels in the treated animals were also lowered (P < 0.05). After 200 ng TRH/rat the increase in TSH was not as high in both groups than after the administration of 100 μg/rat. There was no significant difference between the response of TSH to TRH in the dexamethasone treated and the control rats. The different effects of dexamethasone on PRL and TSH release after TRH may give a further insight into the different regulating mechanisms of both hormones in rats.
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In order to investigate the role of cortisol in the regulation of testicular function, adult male guinea pigs were challenged with ACTH (20 IU), cortisol (8 or 16 mumol), or with ACTH plus dexamethasone (DEX, 2 mumol). The amounts of cortisol, testosterone, progesterone, and androstenedione present in the plasma or secreted by incubated adrenals or testes were determined by radioimmunoassay. The plasma concentrations of LH were determined using a radioimmunoassay for rat LH. ACTH treatment elevated cortisol plasma concentrations to 999% of control values, whereas it reduced testosterone plasma levels to 43% of control values. ACTH treatment did not affect LH plasma levels. A significant negative correlation was found in ACTH-treated animals, when the cortisol and testosterone plasma concentrations in serially taken blood samples (30-240 min after treatment) were compared (rs = -0.90 and rs = -0.99, P < 0.05). In addition to cortisol, ACTH raised progesterone and androstenedione plasma concentrations. If animals were treated with 2 mumol DEX + ACTH, the plasma levels of cortisol and androstenedione but not of progesterone, testosterone or LH were changed. ACTH stimulated the in vitro secretion of cortisol, progesterone and androstenedione by the adrenals but reduced the in vitro release of androstenedione and testosterone by the testes. In summary, treatment of guinea pigs resulted in elevated cortisol and in reduced testosterone plasma concentrations. The mechanism of the cortisol-induced inhibition of testicular function was independent of the LH plasma concentrations. The in vitro experiments indicate that cortisol directly interacts with the Leydig cells, presumably by inhibiting the activity of the testicular 17 alpha-hydroxylase and/or C17,20-lyase. Taking into account the results of comparable investigations in the rat, the inhibition of the testicular 17 alpha-hydroxylase and/or C17,20-lyase takes place if the intracellular cortisol exceeds the capacity of the 11 beta-hydroxysteroid dehydrogenase to inactivate it.
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Detection of hepcidin in myelodysplastic syndrome patients with anemia and its clinical significance
Objective: To study the relationship of Hepcidin and serum ferritin(SF) in myelodysplastic syndrome(MDS) patients with anemia and normal control group,in order to provide the theoretical basis for clinical diagnosis and treatment of iron disorders related anemia.Methods: EnzymE-linked immunosorbent assay(ELISA) and radioimmunoassay were used to measure serum Hepcidin of 45 cases of MDS patients with anemia and 60 cases of normal control group,serum ferritin levels were also detected.Results: Hepcidin levels in the normal control group had nothing to do with age.Hepcidin of male group was significantly higher than that of female group(P0.01),and there was a strong positive correlation between Hepcidin and SF level(r=0.895,P0.01).The Hepcidin level of MDS patients was significantly higher than that of control group(P0.01);for MDS group,the level of Hepcidin and SF in more than 60-year-old patient group was significantly higher than its level in less than 60-year-old patient group(including 60 years old patients,P0.01);the level of Hepcidin and SF in the group of patients receiving blood transfusion more than three times was significantly higher than that in the patients with blood transfusion less than three times(including three times,P0.01).Strong positive correlation was found between Hepcidin and SF levels(r=0.983,P0.01).Conclusion: In physiological circumstances,Hepcidin was mainly regulated by SF,Hepcidin of male was significantly higher than that of females.Hepcidin in patients with MDS was significantly higher than in normal people,and significantly correlated with the number of blood transfusion and age.There was a strong positive correlation between Hepcidin and SF levels in MDS group.
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We have developed a sensitive homologous radioimmunoassay for mouse prolactin (mPRL) capable of measuring concentrations of PRL as low as 1 ng/ml of mouse serum and studied PRL secretion in mice under several physiological conditions. Newborn mice of C3H/St strain had low concentration of PRL in sera at 2–5 days of age which increased with age. Adult females of C3H/St strain contained significantly more PRL in their pituitary glands (7.9 μg/mg) than comparable females of C57BL/St strain (5.6 μg/mg); in contrast, however, they had lower PRL levels in sera (26.6 ng/ml) than the C57BL/ St females (84.5 ng/ml). The females of both strains possessed greater concentration of PRL in their pituitary glands, but in sera only C57BL/St females showed higher levels than males. Castration of C3H mice reduced pituitary PRL in both sexes but decreased serum PRL in males only. Stilbestrol in the diet of castrated mice, however, enhanced pituitary and serum PRL levels in either sex. Adult C3H mice had depressed serum concentrations of PRL following the injection of L-dopa and elevated levels after epinephrine. (Endocrinology91: 1045, 1972)
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