Follicular development and a single ovulation induced with pulsatile administration of Gn-RH in anovulatory women: studies of hormonal analysis and follicular sonometry

The method of pulsatile administration of gonadotropin-releasing hormone (Gn-RH) has been proven as a useful means for induction of ovulation in anovulatory women. In our series of clinical trials, 23 out of 29 anovulatory patients ovulated with pulsatile administration of Gn-RH. Seven patients who ovulated volunteered for the present study with daily hormonal analysis and follicular sonometory . Two patients have oligomenorrhea, 3 patients secondary amenorrhea-1st grade (the sole administration of gestagen required for withdrawal bleeding) and the remaining 2 patients secondary amenorrhea-2nd grade (the combined administration of estrogen and gestagen required for withdrawal bleeding). A diagnosis of hyperprolactinemia was made for one patient with secondary amenorrhea-1st grade. Pulsatile administration of Gn-RH was performed by the use of a self-administered infuser . The infuser was connected to an i.v. indwelling catheter via a specially designed blood backflow eliminater . Five micrograms or less of Gn-RH was given every 2 hr from 07:00 to 23:00 hr daily. Five patients received HCG during the preovulatory period. In one patient, a short term treatment of HMG was added to Gn-RH treatment. Follicular sonometry revealed the development of a single dominant follicle which reached between 20 and 28 mm (23.7 +/- 0.12 mm, mean +/- S.E.) in diameter at the preovulatory period. Disappearance of a dominant follicle was recognized in the early luteal phase. Characteristic increases in estradiol were recognized concomitantly with the development of a dominant follicle. Progesterone levels after ovulation were within the limits of its normal "luteal phase" rise. The present data suggest that pulsatile administration of low dose Gn-RH with nocturnal interruption of treatment is effective for normal progress of follicular development in various types of anovulatory patients, culminating in single ovulation. This paper includes the discussion on our method which may be responsible for a high success rate of ovulation induction.     

Corticotropin-releasing factor in humans. II. CRF stimulation in patients with diseases of the hypothalamo-pituitary-adrenal axis

A stimulation test with 100 micrograms ovine or human corticotropin-releasing factor (CRF) is a useful diagnostic tool in diseases of the hypothalamo-pituitary-adrenal axis. No serious side effects were observed during the test procedure. The results showed that the CRF test is useful in making the differential diagnosis of established Cushing's syndrome (n = 42). The CRF test was also repeated after transsphenoidal surgery in 25 patients with Cushing's disease. Successfully operated patients exhibit no, blunted or normal adrenocorticotropic hormone (ACTH) responses to CRF (n = 15), whereas patients who did not show remission remained hyperresponsive (n = 10). In patients with autonomous adrenal cortisol secretion, the ACTH response to CRF was suppressed (n = 10). After surgery the ACTH response to CRF can already be demonstrated when cortisol levels are still undetectable. Pulsatile administration of CRF in one patient after unilateral adrenalectomy and another patient under corticoid therapy revealed that ACTH responses to CRF normalize rapidly but cannot be sustained if CRF administration is withdrawn, suggesting that the cause of adrenal failure after unilateral adrenalectomy for Cushing's syndrome or long-term corticoid therapy is due to hypothalamic CRF deficiency. The decrease of the ACTH responses to CRF in glucocorticoid-treated patients correlated directly to the daily corticoid dosage. Since the ACTH hyperresponse to CRF in 6 patients with Cushing's disease was also suppressed by short-term dexamethasone treatment, the pituitary level as target site for the acute feedback inhibition is also demonstrated. The evaluation of the CRF-induced ACTH response in patients with secondary adrenal failure without detectable pathology in the sella and suprasellar region (n = 6) enables the differentiation between hypothalamic and pituitary adrenal insufficiency. In patients with hypothalamic lesions the ACTH response to CRF was normal whereas insulin hypoglycemia failed to induce an ACTH rise.     

Effects of streptozotocin-induced experimental diabetes on the morphology and function of the zona fasciculata of rat adrenal cortex

The effects of a severe streptozotocin (STZ)-induced diabetes on the morphology and function of the adrenal zona fasciculata were examined in rats with intact or pharmacologically interrupted hypothalamic-hypophyseal-adrenal axis. In animals with an intact hypothalamic-hypophyseal axis, STZ-diabetes induced hypertrophy of the cells of the zona fasciculata and a rise in the plasma corticosterone concentration. Conversely, in rats in which the hypothalamic-hypophyseal axis had been interrupted, experimental diabetes provoked atrophy of the zona fasciculata cells, and a lowering in the plasma corticosterone level. The effects of STZ-diabetes were completely reversed by insulin infusion in both groups of rats. The hypothesis is discussed that the chronic lack of insulin may directly inhibit the growth and steroidogenic capacity of the rat zona fasciculata and that this effect of experimental diabetes may be masked in rats with an intact hypothalamic-hypophyseal axis by the concurrent enhancement of ACTH release due to chronic stress resulting from the metabolic consequences of prolonged diabetes.     

痛风性关节炎急性期、缓解期超声特征及与血清IL-22、IL-37水平的相关性研究

摘要 目的：探讨痛风性关节炎（GA）急性期、缓解期超声特征及与血清白细胞介素（IL）-22、IL-37水平的相关性。方法：选取从2019年7月至2021年3月期间，在我院诊断为GA的患者64例作为痛风组，及同期在我院体检的健康志愿者32例作为正常对照组。经过超声检查、血清样品实验后，观察受试者的超声特征，比较受试者血清IL-22、IL-37的水平，采用Spearman检验分析GA患者血清IL-22、 IL-37水平与超声特征的相关性。比较超声特征、IL-22和IL-37水平单一或联合应用对GA急性期、缓解期的诊断价值。结果：痛风组急性期血清IL-22水平显著高于缓解期及正常对照组(P＜0.05)；痛风组急性期及缓解期血清IL-37水平均显著高于正常对照组，且缓解期高于急性期(P＜0.05)。64例GA患者中有29例存在痛风石；60例在急性期存在关节积液，至缓解期有22例吸收至正常积液量；有52例在急性期关节滑膜增生，至缓解期有14例得以缓解;有29例存在"双轨征"，仅1例在缓解期消失;有14例发生骨侵蚀，在缓解期均未见明显改善；缓解期积液厚度、滑膜厚度均明显小于急性期(P＜0.05)。无论急性期还是缓解期，GA患者血清IL-22水平与关节积液均呈正相关(P＜0.05)。超声检查联合血清IL-22、IL-37水平检测用于诊断无论在急性期还是缓解期，其敏感性及特异性均较高。结论：IL-22水平不论在急性期还是缓解期，与超声下关节积液均呈正相关。血清IL-22、IL-37检测与超声检查联合可提高急性期及缓解期GA的诊断效能。     

Autologous responses to human leukaemic cells in mixed leucocyte culture

Summary Cryopreserved leukaemic blasts and remission non-T cells from 22 patients with acute leukaemia (15 lymphocytic, 7 non-lymphocytic) were tested as stimulators of autologous remission T cells and normal allogeneic T cells in primary and secondary MLC. In most cases the autologous response elicited by leukaemic cells was less than or equal to that elicited by remission non-T cells. However, T cells from 2 patients in long-standing first remission from ANLL displayed greater proliferation in response to leukaemic blasts than to remission non-T cells in both primary and secondary MLC. The results are suggestive of sensitization of these 2 patients to leukaemia-specific antigens, but other possible explanations are discussed. Abbreviations used: MLC, mixed leucocyte culture; ANLL, acute non-lymphocytic leukaemia; ALL, acute lymphoblastic leukaemia; AMLR, autologous mixed lymphocyte reaction; NK cells, natural killer cells; MNC, mononuclear cells     

An analytical biomarker for treatment of patients with recurrent B-ALL after remission induced by infusion of anti-CD19 chimeric antigen receptor T (CAR-T) cells

Anti-CD19 chimeric antigen receptor-modified T (CAR-T-19) cells have emerged as a powerful targeted immunotherapy for B-cell lineage acute lymphoblastic leukemia with a remarkable clinical response in recent trials. Nonetheless, few data are available on the subsequent clinical monitoring and treatment of the patients, especially those with disease recurrence after CAR-T-19 cell infusion. Here, we analyzed three patients who survived after our phase I clinical trial and who were studied by means of biomarkers reflecting persistence of CAR-T-19 cells in vivo and predictive factors directing further treatment. One patient achieved 9-week sustained complete remission and subsequently received an allogeneic hematopoietic stem cell transplant. Another patient who showed relapse after 20 weeks without detectable leukemia in the cerebrospinal fluid after CAR-T-19 cell treatment was able to achieve a morphological remission under the influence of stand-alone low-dose chemotherapeutic agents. The third patient gradually developed extensive extramedullary involvement in tissues with scarce immune- cell infiltration during a long period of hematopoietic remission after CAR-T-19 cell therapy. Long-term and discontinuous increases in serum cytokines (mainly interleukin 6 and C-reactive protein) were identified in two patients (Nos. 1 and 6) even though only a low copy number of CAR molecules could be detected in their peripheral blood. This finding was suggestive of persistent functional activity of CAR-T-19 cells. Combined analyses of laboratory biomarkers with their clinical manifestations before and after salvage treatment showed that the persistent immunosurveillance mediated by CAR-T-19 cells would inevitably potentiate the leukemia-killing effectiveness of subsequent chemotherapy in patients who showed relapse after CAR-T-19-induced remission.     

Gonadal function in men treated for acute leukaemia.

Gonadal function was assessed in eight men in remission of leukaemia who had completed treatment eight months to eight years previously. All four men treated for acute myeloid leukaemia had normal sperm counts and motility, compared with only one of the four with acute lymphatic or undifferentiated leukaemia. Hormonal studies indicated that the sterility resulted from gonadal failure rather than pituitary dysfunction after cranial irradiation. These findings are important in the counselling of patients with leukaemia.     

Reproductive axis response to repeated lipopolysaccharide administration in peripubertal female rats

Immune system disorders are often accompanied by alterations in the reproductive axis. Several reports have shown that administration of bacterial lipopolysaccharide (LPS) has central inflammatory effects and activates cytokine release in the hypothalamus where the luteinizing hormone releasing hormone (Gn-RH) neurons are located. The present study was designed to investigate the effect of repeated LPS administration on the neuroendocrine mechanisms of control of the reproductive axis in peripubertal female rats (30-day-old rats). With this aim, LPS (50 μg/kg weight) was administered to the animals during 25, 27 and 29 days of age and sacrificed on 30 day of life. Gn-RH, γ?amino butyric acid (GABA) and glutamic acid (GLU), two amino acids involved in the regulation of Gn-RH secretion, hypothalamic content were measured. LH and estradiol serum levels were also determined and the day of vaginal opening examined. The results showed a significant increase in Gn-RH and GLU content (p?<?0.0001), shared by a reduction of GABA one (p?<?0.0001). LH and estradiol serum levels were decreased (p?<?0.01, p?<?0.001) and delay in the day of vaginal opening was also observed in treated animals. Present results show that repeated LPS administration impaired reproductive function, modifying the neuroendocrine mechanisms of control of the axis in peripubertal female rats.     

Chromosomes in acute nonlymphocytic leukemia

Summary The karyotype of leukemic cells was studied in 88 acute nonlymphocytic leukemia (ANLL) patients. Chromosome abnormalities were discovered in 78.4% of all patients and in 72.5% of the 69 patients studied before treatment. Characteristic abnormalities: translocations 8;21, 15;17, 9;22 or 6;9, rearrangements of 11q, gain of chromosomes 8 or 21, and loss or deletion of chromosomes 5 or 7 were detected in 56 of 69 patients with abnormal karyotypes. Translocation 8;21 was revealed in 27 patients; 20 of them had M2 FAB-form, four had M1, and three had M4. In patients with t(8;21) the incidence of complete remission was higher and the duration of first remission and survival longer than in patients with other abnormalities or with a normal karyotype.     

Assessment of the sexual and antler potential of the male white-tailed deer (Odocoileus virginianus) by Gn-RH stimulation test

Twelve mature white-tailed bucks were injected with gonadotropin regulating hormone (Gn-RH, 100 micrograms/deer) during the rut (November) and during the spring (April). In the rut, superior bucks (with actual or potential large body weight, trophy antlers and a high social rank) responded to Gn-RH with a small increase of LH (below 20 micrograms/ml) and a profound rise in testosterone (T) (30-50 ng/ml). The inferior animals exhibited high increase of LH (30-40 ng/ml) but a low rise in T (below 10 ng/ml). FSH levels increased only slightly after Gn-RH and the concentrations were not related to reproductive performance. During the spring, increase in LH levels after Gn-RH administration greatly exceeded the rise of T, but no relationship was found between hormonal levels and the reproductive potential. FSH levels increased remarkably after Gn-RH administration. Gn-RH (administered during the rut) might be used for assessment of the potential for reproductive and antler performance.     

Response to and production of interleukin 2 by peripheral blood and cerebrospinal fluid lymphocytes of patients with multiple sclerosis

In an effort to further characterize the defective proliferative response of T lymphocytes to mitogens in multiple sclerosis (MS) patients, we examined the response to and production of interleukin 2 (IL 2) by both peripheral blood lymphocytes (PBL) and cerebrospinal fluid mononuclear cells. We also examined the proportion of cells bearing receptors for IL 2 and transferrin. Chronic progressive MS patients have an abnormally low response to exogenous IL 2 as compared to controls. Whereas acute relapse patients' PBL demonstrated a normal IL 2 response during an exacerbation, they showed reduced responsiveness during remission. These abnormalities could not be explained by different dose or kinetic response optima to PHA or IL 2, nor could they be explained by depressed numbers of IL 2 or transferrin receptor-bearing lymphocytes. Production of IL 2 by PBL was also abnormal in MS patients. Chronic progressive patients produced elevated levels of IL 2, whereas acute relapse patients undergoing an exacerbation produced diminished levels of IL 2. During remission, these levels returned to that of controls'. The effect of 1200 rad x-irradiation or nylon wool removal of adherent cells was a significantly greater augmentation of IL 2 production in MS patients than in other neurologic disease or normal controls. Cerebrospinal fluid lymphocytes from MS patients had normal proportions of IL 2 receptor-bearing cells, but were deficient in their IL 2 response and production as compared to autochthonous or control PBL. The inability of some MS patients' lymphocytes to clonally expand in response to IL 2 might contribute to the pathogenicity of the disease.     

Gonadotropin response to slow infusion of GN-RH in idiopathic ologospermia. Preliminary findings

Gn-RH was administered i.v. to infertile patients presenting with idiopathic oligospermia at a constant rate (0,4 microgram) for 4 h. All the patients showed an increased FSH response, and LH levels higher than normal in both the first and the second peak of response. Our data indicate that an increased pituitary sensitivity and reserve of FSH and LH are present in idiopathic oligospermia, suggesting an involvement of both Leydig cells and seminiferous epithelium in this condition.     

The acute stimulatory effect of estrogen on gonadotropin release in gonadotropin-releasing hormone-primed women

In order to prove the acute stimulatory effects of estrogen on pituitary gonadotropin release, we have performed the present experiments in 8 women with a hypergonadotropic state due to surgical castration or primary ovarian failure. They received gonadotropin releasing hormone (Gn-RH) for 12-21 h at the constant rate of 20 micrograms/h. In 5 of the women, estradiol-17 beta was concomitantly administered at the rate of 20 micrograms/h from 6 h after the start of Gn-RH infusion. Blood samples were collected frequently throughout the experiments for the analysis of LH, FSH and estradiol. In response to the sole stimulation of Gn-RH, remarkable and prompt rises in LH (313.5%), but to a lesser degree in FSH (194.2%), were observed within the initial 3 h, and their high levels were maintained throughout the experimental period. However, the additional administration of estradiol brought on a further sudden rise in both gonadotropins levels: 178.3% for LH and 163.5% for FSH within 2 h. These high levels were sustained during estradiol infusions. In 2 of them, blood samples were obtained for several hours after cessation of estradiol infusion. The circulating gonadotropin level dropped precipitously close to the baseline level within 3 h after estradiol infusions. Our data indicate that estrogen has an acute and strong augmentative effect on Gn-RH induced gonadotropin release in addition to its conventional negative and positive feedback effects.     

Induction of ovulation with chronic intermittent (pulsatile) administration of Gn-RH in women with hypothalamic amenorrhoea

The physiological and pathophysiological basis of hypothalamic amenorrhoea are reviewed as well as the clinical results of chronic intermittent (pulsatile) administration of Gn-RH in the treatment of infertility. Hypothalamic amenorrhoea is considered to be the result of a deficient hypothalamic secretion of Gn-RH. By pulsatile administration of Gn-RH, which is a pre-requisite of normal pituitary gonadotrophic function, deficient endogenous Gn-RH is replaced. If an adequate dose of Gn-RH is provided, which takes into account the degree of impairment of hypothalamic function in the individual case, follicular maturation, ovulation and corpus luteum formation are achieved in nearly every treatment cycle. Although dependent also on factors other than the treated dysfunction, a high conception rate is achieved.     

IL17 and IFNgamma production by peripheral blood mononuclear cells from clinically isolated syndrome to secondary progressive multiple sclerosis

We evaluated the spontaneous IL17, IFNgamma and IL10 production by peripheral blood mononuclear cells from patients affected by clinically isolated syndromes (CIS) suggestive of multiple sclerosis (MS) both in acute phase and in remission, relapsing remitting MS (RRMS) both in relapse and in remission, not-relapsing secondary progressive MS (SPMS) and controls. We observed higher IL17 levels in CIS patients both in acute phase and in remission than in SPMS patients and controls. On the contrary no difference in IL17 production was observed among RRMS patients and CIS, SPMS patients and controls. IFNgamma levels were significantly higher in CIS patients in acute phase than in CIS and RRMS patients in remission, SPMS patients and controls. Moreover, we observed higher IFNgamma spontaneous production in relapsing RRMS patients than in remitting RRMS and SPMS patients and controls. IL10 levels were significantly higher in remitting CIS and in relapsing RRMS patients than in SPMS patients and controls. There was no difference in IFNgamma, IL10 and IL17 levels between SPMS patients and controls. Our data suggest that IL17 might play a crucial role mainly in the early phase of MS, while IFNgamma seems to be involved both in the early phase and in the following relapses of the disease.     

Probable gamma-aminobutyric acid involvement in bisphenol A effect at the hypothalamic level in adult male rats

The aim of the present study was to investigate the effects of bisphenol A (BPA) on the neuroendocrine mechanism of control of the reproductive axis in adult male rats exposed to it during pre- and early postnatal periods. Wistar mated rats were treated with either 0.1% ethanol or BPA in their drinking water until their offspring were weaned at the age of 21 days. The estimated average dose of exposure to dams was approximately 2.5 mg/kg body weight per day of BPA. After 21 days, the pups were separated from the mother and sacrificed on 70 day of life. Gn-RH and gamma-aminobutyric acid (GABA) release from hypothalamic fragments was measured. LH, FSH, and testosterone concentrations were determined, and histological and morphometrical studies of testis were performed. Gn-RH release decreased significantly, while GABA serum levels were markedly increased by treatment. LH serum levels showed no changes, and FSH and testosterone levels decreased significantly. Histological studies showed abnormalities in the tubular organization of the germinal epithelium. The cytoarchitecture of germinal cells was apparently normal, and a reduction of the nuclear area of Leydig cells but not their number was observed. Taken all together, these results provide evidence of the effect caused by BPA on the adult male reproductive axis when exposed during pre- and postnatal period. Moreover, our findings suggest a probable GABA involvement in its effect at the hypothalamic level.     

Oestrus, time of ovulation, ovulation rate and conception rate in progestagen-treated ewes given Gn-RH, Gn-TH analogues and gonadotrophins.

The influence of Gn-RH, hCG and a PMSG-hCG mixture (PG600) on the time of ovulation, ovulation rate and on the occurrence of oestrus in ewes treated with progestagen-impregnated sponges for 12 days examined. The effects of Gn-RH analogues on plasma LH, oestrus, ovulation and conception rate were also investigated. Six separate experiments were carried out. When 50 micrograms Gn-RH were given 24 h after sponge removal ovulation occurred in 44--46% of ewes within 24 h and in all ewes by 34 h. Gn-RH was a more potent ovulation synchronizer than hCG. Both hCG and PG600 reduced the incidence of overt oestrus. Gn-RH also had this effect in ewes treated during February and May but not in August and September. Gn-RH analogues given 2 days before sponge removal significantly increased ovulation rate. The display of oestrus was not affected in ewes treated 2 days before sponge removal but was suppressed in 43-69% of ewes treated with an analogue at the time of sponge removal. Ovulation occurred in 50-62% of ewes within 30-35 h of injection of Gn-RH analogues, regardless of the time of their administration. The release of LH in response to one analogue was not influenced by the presence of the progestagen-impregnated sponge in the vagina. When given a Gn-RH analogue 2 days before sponge removal or at the time of sponge removal 63 and 62% of mated ewes became pregnant compared with 70% of control ewes.     

The study of cell-mediated immune response in recurrent vulvovaginal candidiasis

The aim of this work was to examine in vitro the ability of cells from patients with recurrent vulvovaginal candidiasis (RVVC) to cell-mediated immune response. Peripheral blood mononuclear cells (PBMC) and whole blood cells (WBC) of 37 RVVC patients in acute infection and 14 in remission were examined for the ability to proliferation and cytokines production (IFN, TNF, IL-6). As a control, a group of 25 healthy women were examined. The cells were stimulated with Candida antigen (HKCA), LPS and PHA. To indicate the level of cytokines, the following cell-lines were used: A549 for IFN, WEHI 164 for TNF and 7TD1 for IL-6. The proliferation/death of cells was determined by colorimetric test using MTT. Distinct suppression of cell-mediated immune response (CMI) was shown in all patients comparing to the control. Greatest suppression was found in the acute phase of the disease. The ability of cells to proliferate and produce IFN increases only in remission. The data seem to suggest that in this phase of disease, the ability of cell-mediated immune response is restored. It was also indicated that IFN may take part in protection against Candida infection.     

Zinc level in the blood of children with nephrotic syndrome and correction of its deficiency with animal blood preparation Livex]

Serum and erythrocyte zinc levels have been assayed in in 45 children with steroid-dependent nephrotic syndrome in both acute phase and remission. Out of these children a group of 22 has been distinguished. These patients have been treated with Livex--animal blood preparation containing amino acids and trace elements, including zinc. It was found, that serum zinc has been significantly lowered in children with the acute phase of nephrotic syndrome who were not treated with Livex. Erythrocyte zinc levels have been normal in these patients. A significant increase in both serum and erythrocyte zinc levels has been noted during remission, but they have still been lower that in healthy children. A three-month cure with Livex produced statistically significant increase in zinc levels in children during remission.     

Effect of GnRH administration on blood polyamines and LH levels in normal and obese children

Blood polyamines (spermidine and spermine) and LH levels have been studied after acute GnRH injection both in obese and normal weight children. In both groups LH values significantly increased after stimulation but reached higher peaks in normal children than in obese ones (P less than 0.05). On the contrary, polyamine levels increased significantly only in the normal weight children. LH peaked at 30 min and polyamines at 60 min after GnRH injection. On the basis of the proposed role of polyamines in hormone action and of our results, we suggest that polyamines may play a pivotal role in hormone responsiveness of hypothalamic-hypophyseal axis.