Effect of gonadotropin-releasing hormone analog on ovarian reserve in children with central precocious puberty.

BACKGROUND: Gonadotropin-releasing hormone analog (GnRHa) is the mainstream treatment for central precocious puberty (CPP). However, its effect on the ovarian reserve in CPP girls remains unclear. This study was designed to analyze the changes of ovarian reserve in CPP girls during and after GnRHa therapy, with an attempt to achieve the early prediction of the effect of GnRHa treatment on the reproductive function of CPP girls, eliminate the concerns of girls and their parents on the potential toxicities of GnRHa treatment, and improve the patients' adherence to treatment. METHODS: The clinical data of 383 CPP girls who had been treated with GnRHa for more than half a year in our hospital within the past 10 years were retrospectively analyzed. The serum luteinizing hormone (LH), follicle-stimulating hormone (FSH), FSH/LH, estradiol (E2), and anti-Müllerian hormone (AMH) levels, as well as uterine and ovarian volumes, were measured before treatment, at various time points during treatment, and after menarche or resumption of menses (ROM) after treatment discontinuation. RESULTS: GnRHa treatment had similar effects on uterine/ovarian volumes, LH, FSH, and E2: these indicators were significantly inhibited during the treatment (compared with the pre-treatment levels), gradually returned normal after drug withdrawal, and became significantly higher than the pre-treatment levels after menarche or ROM (both P<0.05 for LH and FSH levels and P>0.05 for E2 and uterine/ovarian volumes). AMH level transiently decreased 6 months after GnRHa treatment (2.70±1.76 vs. 3.56±2.21, t=3.227, P=0.001); however, the AMH levels after 12, 18, and 24 months of treatment were similar to the pre-treatment level (P>0.05). The FSH/LH ratio significantly increased after 12 months of treatment compared with the pre-treatment (P<0.05), and the FSH/LH ratio after menarche or ROM was significantly lower than the pre-treatment value (1.34±0.66 vs. 5.69± 6.85, t=3.068, P=0.006). When FSH/LH and FSH level were used to reflect the ovarian reserve, the proportion of CPP girls with normal ovarian reserve after menarche or ROM was higher than at pre-treatment (FSH/LH ratio: 100% vs. 46%, χ2=27.586, P<0.05; FSH level: 100% vs. 99%, P>0.05). When AMH level was used to reflect the ovarian reserve, the proportion of CPP girls with normal ovarian reserve after menarche or ROM was slightly lower than at pre-treatment (87% vs. 93%, P>0.05). CONCLUSIONS: The ovarian reserve of CPP girls is somehow inhibited during GnRHa treatment but is gradually restored after drug discontinuation. Thus, GnRHa treatment does not affect ovarian reserve in CPP children after the treatment stops.

[Effects of HPGA suppression on predicted adult height in girls with central precocious puberty].

OBJECTIVE: To study the relationship between the suppression of the hypothalamic-pituitary-gonadal axis (HPGA) and the predicted adult height (PAH) in girls with central precocious puberty (CPP) during the treatment with gonadotropin-releasing hormone analogue (GnRHa), in order to provide guidance for individualized GnRHa dose adjustment in clinical practice. METHODS: The clinical data of 75 CPP girls were collected, and then height, bone age (BA), uterine and ovarian volumes, and peak luteinizing hormone (LH), peak follicle-stimulating hormone (FSH), and estradiol (E2) levels were recorded at different time points of GnRHa treatment. PAH at each time point was calculated. PAH improvement (ΔPAH=PAH-target height) and its relationship with the degree of HPGA suppression were analyzed. Threshold effect analysis was applied to determine the best HPGA suppression range forΔPAH. RESULTS: After GnRHa treatment, PAHs were improved markedly compared with the data in the early stage of treatment. ΔPAH showed a negative correlation with ΔBA. At 24 months of treatment, ΔPAH was also negatively correlated with LH. Uterine volume controlled between 2.3 and 3.0 mL, LH level controlled below 0.8 IU/L, and FSH controlled below 2.4 IU/L could slow down the growth of BA and improve PAH. CONCLUSIONS: GnRHa treatment can improve the PAH of CPP girls. Selection of an appropriate therapeutic dose for GnRHa to control uterine volume, LH and FSH levels within certain ranges can slow down the growth of BA and improve PAH.

Lateral ventricle injection of orexin-A ameliorates central precocious puberty in rat via inhibiting the expression of MEG3.

BACKGROUND: Central precocious puberty (CPP) is characterized as increasing gonadotropin-releasing hormone (GnRH) release. Orexin-A has also been shown to affect GnRH release. However, there are few reports about the effect of orexin A on the treatment of CPP. METHODS: After establishing the precocious puberty model, the rats were divided into four groups: normal control, precocious puberty rats, precocious puberty rats treated with normal saline and precocious puberty rats treated with orexin-A. The vaginal opening time, second estrus cycle, ovarian index and uterus index of rats in each group were detected. qRT-PCR was performed to examine the expression of MEG3 and kisspeptin in rats. HT22 cells were transfected with pcDNA-MEG3 to detect the expression of Kisspeptin. RESULTS: In this study, we found that orexin-A not only delayed the day of vaginal opening and regular estrus cycle days but also decreased the ovarian index and uterus index in rats with CPP. In addition, orexin-A reversed the up-regulation of MEG3 and kisspeptin in rats with CPP. In HT22 cells, the mRNA and protein level of kisspeptin were enhanced by pcDNA-MEG3. CONCLUSION: Our results suggest that orexin-A ameliorates central precocious puberty in rat and MEG3 might be involved in this effect, suggesting that MEG3 might be a novel target in treating central precocious puberty.

Different outcomes of hypospadias surgery between north america, europe and china: is patient age a factor?

UNLABELLED: BACKGROUND AND OBJECTS: The patient's age at the first hypospadias repair may be an important factor for determining postoperative outcomes. Age at the first procedure differs between Western countries and medical centers in China. This review examines the differences between the incidence of surgical complications and surgical age in boys receiving hypospadias repairs in North America, Europe and China. MATERIALS AND METHODS: Literature reports were reviewed in PubMed and WanFang databases using the key terms and phrases; 'hypospadias outcomes', 'complications of hypospadias repair' and 'timing of hypospadias repair'. All peer-reviewed articles published over the past decade (2001-2011) were considered if; a full text was available, the article included age at the first hypospadias procedure and surgical complications. RESULTS: In total, 16 131 patients were reported in 113 papers from North America, Europe and China according to our inclusion criteria. There was a significant difference in age at the first hypospadias surgery (P < 0.0005) and in the incidence of complications (P <0.001) between the different global regions investigated, with the earliest surgeries occurring in North American patients. Urethral fistulas were the most common complication reported in all of the regions included in this study. CONCLUSIONS: Fellowship training in pediatric urology could improve surgical outcomes, particularly in young children. Younger children experience fewer complications following hypospadias surgeries, independent of training and access to resources.

[Study on allelopathy effect of pericarp extract of Phellodendron amurense].

Through the study of allelopathy of the pericarp of Phellodendron amurense, the role of self-regeneration barriers was investigated in order to find ways and means for the protection of wild populations of P. amurense. Solution preparation: soaked pericarp of P. amurense in distilled water at 4 degrees C to get solution A, and reflux extraction of pericarp with distilled water at 100 degrees C to get solution B. Both of the solution A and solution B were used in the experiment of seed germination and seedling growth with the seeds of cabbage and wheat. The results showed that 20 g x L(-1) concentration of solution A and solution B inhibited significantly seed germination of cabbage and wheat, while 100 g x L(-1) concentration of solution A even completely inhibited the seed germination of wheat. 20 g x L(-1) concentration of solution A significantly inhibited the cabbage and wheat seedling growth, completely inhibited the root growth of cabbage, while 100 g x L(-1) concentrations of solution A completely inhibited seedling growth of cabbage and wheat. Comparing to solution A, the intensity of solution B are diminished on seed germination and seedling growth. It is concluded that the allelopathy of pericarp of P. amurense is multi-material role in the results, some of allelochemicals are easily degradable when exposed to heat. Overall, the allelopathy of pericarp of P. amurense can affect the seed germination and seedling growth. It is supposed that allelochemicals existed in the pericarp of P. amurense is one of the reason leading to difficulties in self-regeneration of its population.

Effects of vitamin A deficiency on mucosal immunity and response to intestinal infection in rats.

OBJECTIVE: Vitamin A deficiency is associated with an increased incidence of infectious respiratory and alimentary tract diseases in children, and vitamin A supplementation can prevent and assist in treating these diseases. To clarify the mechanisms of these associations, we investigated the effects of vitamin A deficiency on mucosal immunity to intestinal infection in rats. METHODS: Specific pathogen-free Sprague-Dawley rats received a vitamin A-free diet, with (n=20) or without (n=20) vitamin A supplementation. Intestinal infection was induced by oral inoculation of salmonella in 10 rats in each group. The rats were killed 3 d after infection was induced, and we measured the number, maturation, and activation of dendritic cells; the expression of Toll-like receptors 2 and 4; mRNA level of myeloid differentiation primary response gene (88) (MyD88, pattern-recognition receptors and their adapter protein); immune cytokine production in the intestinal mucosa; and the amount of secretory immunoglobulin A in the gut. RESULTS: In vitamin-A deficient rats, the number of mucosal dendritic cells and the production of IL-12 markedly increased; the mucosal expressions of Toll-like receptor 2 and MyD88 were up-regulated, and secretions of interferon-γ and secretory immunoglobulin A were decreased. Infection aggravated the damage to the intestinal mucosa and lowered immunity in vitamin-A deficient rats. CONCLUSION: Vitamin A deficiency damaged both humoral and cellular immunity in the mucosa. Modulation of dendritic cells is likely an important mechanism through which vitamin A deficiency affects mucosal immune responses against infection.

Expression of retinoic acid receptors in intestinal mucosa and the effect of vitamin A on mucosal immunity.

OBJECTIVE: To explore the mechanism of vitamin A (VA) modulation of mucosal immunity, the expression of retinoic acid receptors in intestinal mucosa was measured, and the effect of VA on intestinal dendritic cells (DCs) and mucosal cytokine production was examined. METHODS: The expression of retinoic acid receptor (RAR-alpha, RAR-beta, RAR-gamma, RXR-alpha, RXR-beta, RXR-gamma) mRNA, the distribution and number of DCs, and the protein secretion of interleukin (IL)-12, T-helper type 1/2 cells (interferon [IFN]-gamma/IL-4), and regulatory (IL-10) cytokines in the mucosa of terminal ileum in normal rats and rats with VA deficiency (VAD) were detected by reverse transcriptase polymerase chain reaction, immunohistochemistry, and enzyme-linked immunosorbent assay, respectively. The effect of all-trans retinoic acid on the number and maturation of DCs and the gene expression of RAR-alpha and cytokines listed earlier in cultured Peyer's patches were examined by flow cytometry and reverse transcriptase polymerase chain reaction, respectively. RESULTS: In the intestinal mucosa of VAD rats, RAR-alpha mRNA was downregulated, DC number increased, the protein secretion of IL-12 was increased, but the secretion of IFN-gamma and IL-10 decreased. In in vitro cultured Peyer's patches, all-trans retinoic acid promoted DC maturation, upregulated RAR-alpha mRNA, reduced IL-12 and IFN-gamma, but increased IL-10 gene expression; these effects of all-trans retinoic acid were reversed when cultured with Ro 41-5253 (a specific antagonist of RAR-alpha). CONCLUSION: Vitamin A may be potent in reducing intestinal inflammation and restoring impaired antibody responses in a VAD situation. The effect of VA on DCs could be an important mechanism contributing to altered mucosal immunity. RAR-alpha may mostly play a role in the action of VA.

Effect of all-trans-retinoic acid on the differentiation, maturation and functions of dendritic cells derived from cord blood monocytes.

We investigated the effects of all-trans-retinoic acid on dendritic cells derived from human cord blood monocytes to clarify how vitamin A affects immune function in children. Monocytes were separated from 18 cord blood samples, and dendritic cells were differentiated by culture. The percentage of dendritic cells was markedly lower in all-trans-retinoic acid treated cells than in untreated cells. After exposure to tumour necrosis factor-alpha for 3 days, all-trans-retinoic acid treated dendritic cells showed a reduced capacity to activate alloreactive T cells compared to untreated cells. In addition, all-trans-retinoic acid-treated dendritic cells could drive T cells towards T-helper cell type 2 responses with decreased secretion of interleukin-12, interferon-gamma, and increased production of interleukin-10 and interleukin-4. However, when Ro 41-5253, a selective retinoic acid receptor alpha antagonist, was add to culture, all the above actions were reversed. Thus, all-trans-retinoic acid may act at the first step of the immune response by inhibiting the differentiation of dendritic cells, maturation and induction of the T-helper cell type-2 response. The actions of all-trans-retinoic acid on dendritic cells were mediated through retinoic acid receptor alpha.

[Effects of vitamin A on the differentiation, maturation and functions of dendritic cells from cord blood].

OBJECTIVE: It is well known that vitamin A can improve mucosal immunity and anti-infection immunity. But the mechanisms thereof remain to be clarified. Previous studies on the role of vitamin A in immune regulation focused on lymphocytes, whereas little had been done about dendritic cells, which play very important roles in immune response. The objective of this study was to understand the effects of retinoic acid (RA), the metabolic product of vitamin A in vivo,on the differentiation, maturation and functions of dendritic cells from cord blood. METHODS: Cord blood samples were collected from nine well-nourished full-term neonates. Mononuclear cells were isolated by Ficoll-Hypaque gradient centrifugation and cultured in the presence of 1000 u/ml GM-CSF, 500 u/ml IL-4 for 6 days, then TNF-alpha 20 ng/ml was added into the medium and cultured for another 3 days. The cells were incubated with or without 1 x 10(-6) MRA. Expression of surface molecules, CD1a, CD83, HLA-DR on DC was measured by flow cytometry. The ability of DC derived from the culture to induce proliferation of T cells in the mixed lymphocyte reaction (allo-MLR) was used for the evaluation of their function. IL-12, IFN-gamma, IL-4 and IL-10 were detected at mRNA levels by RT-PCR to understand the roles of DC treated with RA in regulation of Th1/Th2 balance. RESULTS: On the sixth day of cell culture, the percentage of DC incubated with RA (57.28 +/- 9.22) was much lower than that without RA (79.57 +/- 11.85) (P < 0.001), but on the ninth day, there were no differences between the presence or absence of RA (76.18 +/- 10.27 vs. 73.72 +/- 15.58). When RA was added to the medium and the culture was continued for nine days, the percent of immature DC (CD1a + HLA-DR+) was much higher than that of the control (absence of RA) (58.93 +/- 4.70 vs. 45.80 +/- 7.88, t = 6.575, P < 0.001); whereas, mature DC (CD83 + HLA-DR+) percentage was markedly lower than that of the control (17.25 +/- 8.49 vs. 27.92 +/- 13.94, t = 4.435, P = 0.002). The T lymphocytes proliferation induced by the DC treated with RA was reduced from 16 857 +/- 3 643 to 11 924 +/- 2 576 cpm (t = 5.598, P < 0.001) in allo-MLR. Expression of mRNA for IL-12p35, IL-12p40, IFN-gamma in the cells that had been incubated with RA declined, but IL-10, IL-4 increased significantly. CONCLUSION: Vitamin A inhibited the differentiation and maturation of cord blood DC, reduced it's ability to stimulate allo-T lymphocytes proliferation, and down-regulated Th1 cytokines, up-regulated Th2 cytokines, consequently made immune response inclined to Th2.