[Effect of pulsatile GnRH therapy on pituitary-testicular axis function in male patients with pituitary stalk interruption syndrome].

OBJECTIVE: To evaluate the efficacy of pulsatile gonadotropin-releasing hormone (GnRH) therapy in male patients with pituitary stalk interruption syndrome (PSIS). METHODS: Fourteen male PSIS patients, who had received subcutaneous pulsatile GnRH therapy for at least four weeks at Peking Union Medical College Hospital from April 2014 to September 2015, were included in the study. Serum luteinizing hormone (LH), follicle-stimulating hormone (FSH), testosterone level, testicular volume and sperm in ejaculation were measured. After treatment for four weeks, subjects with serum LH≥1.24 U/L were classified as"good response group", and others were classified as"poor response group". RESULTS: All patients were born by breech delivery and presented with absent pubertal development. Laboratory findings showed combined pituitary hormone deficiency in all patients. Pituitary stalk interruption syndrome was diagnosed based on MRI findings. Totally 11 out of 14 patients showed good response to pulsatile GnRH therapy. Their LH level increased from 0.32 (0.15, 0.61) U/L to 4.34(2.52, 7.39)U/L(P<0.001), FSH from 0.84 (0.30, 1.50) U/L to 7.12 (4.53, 10.09) U/L(P<0.001); serum testosterone level from 0 (0, 0.45) nmol/L to 2.46 (0.56, 11.45) nmol/L (P=0.004) and testicular volume from 2.0 (2.0, 4.0) ml to 4.0 (3.0, 6.0) ml (P=0.005). Sperm was detected in seminal samples after treatment for 8 weeks in two patients and for 12 weeks in one patients. Three out of 14 patients showed poor response to pulsatile GnRH therapy. CONCLUSION: Pulsatile GnRH therapy can effectively establish the pituitary-testicular axis function for most patients with pituitary stalk interruption syndrome, indicating enough reserve of gonadotrophs in the pituitary.

[Dysfunction of hypothalamic-pituitary-testicular axis in patients with adrenal hypoplasia congenita due to DAX-1 gene mutation].

OBJECTIVE: To evaluate the hypothalamic-pituitary-testicular axis (HPTA) function and spermatogenesis in male patients with X-linked adrenal hypoplasia congenita (AHC) due to DAX-1 gene mutation. METHODS: Twenty-four adult male patients from Peking Union Medical College Hospital between November 2007 and December 2014 were included.Their DAX-1 gene mutations were confirmed by polymerase chain reaction (PCR) and clinical features, hormone level and semen assay were collected. RESULTS: All patients presented with adrenal deficiency symptom.21 patients (87.5%) showed the symptoms before the age of 10 years old.The average testicular volume was 2.0 (2.0, 3.8) ml at the baseline in 24 patients.Three patients had cryptorchidism.Serum testosterone level increased from 0.1 (0, 0.5) nmol/L to 13.5 (7.6, 15.4) nmol/L (n=16) after human chorionic gonadotropin (HCG) stimulation.23 out of 24 patients were diagnosed as hypogonadotropic hypogonadism (HH) and only one patient was diagnosed as azoospermia with normal gonadotropin and testosterone level.Combined gonadotropin therapy was administered in seven patients and their serum testosterone level reached 15.3(8.4, 25.3) nmol/L, but no obvious testicular enlargement was observed [(4.0±2.9) vs (4.9±3.3) ml , P=0.270] and seminal analysis revealed persistent azoospermia.Another patient showed response to pulsatile GnRH therapy.Luteinizing hormone(LH) level increased from 1.0 U/L to 9.3 U/L, and follicle stimulating hormone(FSH) level increased from 3.0 U/L to 13.5 U/L.Serum testosterone level increased from 0 nmol/L to 10.0 nmol/L, but testicular volume maintained 3 ml after treatment for two months. CONCLUSIONS: Male patients with DAX-1 gene mutations presented with primary defect in spermatogenesis and hypogonadotropic hypogonadism.Their Leydig cell function was almost normal, while Sertoli cell and seminiferous tubule function were seriously damaged.

Bioactivity-guided fractionation for analgesic properties and constituents of Vitex negundo L. seeds.

This study was undertaken to ascertain the analgesic properties of Vitex negundo L. seeds and to isolate and characterize the active constituents. Among the 80% ethanol extract and some fractions with different polarity, the acetoacetate fraction showed the highest anti-nociceptive activity in acetic acid-induced writhing test in ICR mice. The analgesic bioguided isolation of the acetoacetate fraction yielded two major lignans: 6-hydroxy-4-(4-hydroxy-3-methoxy-phenyl)-3-hydroxymethyl-7-methoxy-3,4-dihydro-2-naphthaldehyde (1) and vitedoamine A (2). Given orally, compound (1), which was more productive, produced significant inhibitions on chemical nociception induced by intraperitoneal acetic acid and subplantar formalin injections and exhibited notable anti-inflammatory activities in dimethyl benzene-induced ear edema test in a dose-dependent manner. Since co-administration of naloxone fails to antagonize the analgesic activity of compound (1) in the formalin test, we suggest that compound (1) possesses potent analgesic effects which are most likely to be mediated by its anti-inflammatory activity rather than through opioid receptor system and therefore could partially explain the anti-nociceptive effect of V. negundo L. seeds.

Evaluating congenital spine deformities for intraspinal anomalies with magnetic resonance imaging.

SUMMARY: The incidence of intraspinal abnormalities associated with congenital spinal anomalies as detected by magnetic resonance imaging (MRI) is becoming better defined. In this study, 41 nonrandomized children with congenital spinal deformities (excluding myelomeningocele) who underwent complete MR evaluation were reviewed. Of the 41 congenital spinal deformities, 37 demonstrated congenital scoliosis, with failure of formation in 19, failure of segmentation in 4, and mixed defects in 14. The remaining four deformities were cases of congenital kyphosis. Thirteen patients with congenital spine anomalies were noted to have intraspinal abnormalities identified by MRI: tethered cord in 12 patients, syringomyelia in 3 patients, and diastematomyelia in 5 patients. Of the 12 patients with tethered cord, 2 patients had neurologic deficits. Urorectal anomaly was one of the most common associated findings (15%). Considering an incidence of intraspinal anomalies of 31% and as clinical manifestations may not be initially detectable, MRI is recommended in patients with congenital spinal deformity as part of the initial evaluation even in the absence of clinical findings.

Insulin-like growth factor I production is essential for anabolic effects of thyroid hormone in osteoblasts.

Thyroid hormone (T3) and insulin-like growth factor I (IGF-I) are critical regulators of skeletal function. T3 increases IGF-I production in bone. To assess the potential role of IGF-I as a mediator of T3 actions, we characterized phenotypic markers of osteoblast activity in two osteoblast models, normal mouse osteoblasts and MC3T3-E1 cells, exposed to T3 alone or under conditions that interfere with IGF-I actions. T3 significantly increased osteoblast 3H-proline incorporation, alkaline phosphatase (ALP), and osteocalcin. Both alphaIR3, a neutralizing monoclonal antibody to the IGF-I receptor, and JB1, an IGF-I analogue antagonist, attenuated the stimulatory effects of T3. T3 effects also were decreased in cells transfected with antisense oligonucleotide (AS-ODN) to the IGF-I receptor gene. Both IGF-I and T3 had mitogenic effects that were inhibited by the antagonists. IGF-I by itself did not stimulate 3H-proline incorporation, ALP, and osteocalcin in the models used, revealing that although IGF-I is essential for the anabolic effects of T3, it acts in concert with other factors to elicit these phenotypic responses.

Resolution of the basal plasma membrane calcium flux in vascular smooth muscle cells.

Steady-state cytosolic calcium (Ca2+i) concentration in a vascular smooth muscle cell is determined by Ca2+ influx and Ca2+ extrusion across the plasma membrane, yet no means for determining the absolute magnitude of these transmembrane Ca2+ fluxes in the basal state of the resting cell has been devised. We now report a method that combines fluorescence measurement of Ca2+i, 45Ca kinetics, and computer modeling to yield the basal plasma membrane Ca2+ flux in A7r5 vascular smooth muscle cells. Kinetic analysis of basal Ca2+i and Ca2+i transients following chelation of extracellular Ca2+ yields a unique value for the ratio of the rate constant governing Ca2+ pumping into the sarcoplasmic reticulum (SR) to that for plasma membrane Ca2+ extrusion (1.12 +/- 0.06). When this ratio was used to constrain the least-squares fitting of 45Ca efflux data from A7r5 cells, it was possible to determine unique values for the unidirectional, steady-state Ca2+ fluxes across both SR and plasma membranes. The basal unidirectional plasma membrane Ca2+ flux was 0.062 +/- 0.018 fmol . min-1 . cell, and the basal SR Ca2+ flux was 0.069 +/- 0.019 fmol . min-1 . cell-1. These results demonstrate, within the limitations of measuring the absolute value of Ca2+i, the feasibility of measuring previously unresolvable subpicoamp basal Ca2+ fluxes in intact cells under normal physiological conditions.

Mechanisms for Ca signaling in vascular smooth muscle: resolved from 45Ca uptake and efflux experiments.

Established cell lines are now widely used in experiments concerning vascular smooth muscle (VSM) function; however, considerable evidence suggests that cultured VSM cells are functionally different from VSM cells in intact blood vessels. In order to test the hypothesis that calcium signaling mechanisms are comparable in these two preparations, we developed a new method for high resolution 45Ca efflux studies in A7r5 cells. Briefly, this method involves plating cells in the lumen of a tubular glass efflux chamber and, after loading the cells with 45Ca, perfusing the chamber with a physiological saline solution and collecting the effluent. Using this method we found that the plasma membrane in cultured cells is not rate limiting for calcium efflux, since the efflux curves from both permeabilized and intact cells are kinetically the same. We also found the plasma membrane is not rate limiting in whole aortic segments by using a depolarizing solution followed by dihydropyridine solution. Thus, we demonstrated that the data obtained from cells or tissues with intact membranes reveal information about the intracellular stores (sarcoplasmic reticulum and mitochondria). Combining efflux data with a detailed kinetic model of cellular Ca transport allows least-squares estimation of the rate constants for release and uptake of Ca2+ by intracellular stores with a high degree of confidence (CV < 25%) as well as the Ca2+ contents and transmembrane fluxes associated with these stores. Quantitative comparison of results obtained from A7r5 cells with those we previously obtained for rabbit aortic segments reveals marked similarities and suggests that A7r5 cells serve as excellent model experiments for VSM cell Ca2+ homeostasis.

[Comparison between essential oils and amino acids in fresh and processed roots of Changium smyrnioides Wollf].

This paper deals with a comparative study between essential oil and amino acid constituents from the fresh root and processed product of Changium smyrnioides. Twenty-seven ingredients were identified from the essential oil, and the amino acids in roots pro- and postprocessed were found varying greatly in content.