OBJECTIVE

This study was designed to compare the serum levels of fibroblast growth factor 23 (FGF23) among patients with gestational diabetes mellitus (GDM) and healthy pregnant women, and to evaluate the association between hormonal and metabolic parameters.

METHODS

A total of 82 pregnant women were consecutively enrolled in the study. Of these, 46 were diagnosed as having GDM; the remaining 36 healthy pregnant women served as controls in a cross-sectional study design. The womens' ages ranged from 22 to 38 years and gestational ages, from 24 to 28 weeks. Serum samples were analyzed for FGF23 levels using an enzyme-linked immunosorbent assay.

RESULTS

Serum FGF23 levels were increased in patients with GDM compared with controls (median, 65.3 for patients with GDM vs. 36.6 ng/mL for healthy controls; p = 0.019). Mean fasting glucose (105.6 ± 7.4 vs. 70.2 ± 7.2 mg/dL, p < 0.001), HbA1c (5.6 ± 0.5 vs. 4.9 ± 0.5%, p < 0.001), insulin (median, 11.1 vs. 8.7 µIU/mL, p = 0.006) and HOMA-IR (3.0 (1.8) vs 1.4 (0.6), p < 0.001) levels were significantly higher in patients with GDM than in controls. Serum FGF23 level was positively correlated with body mass index (r2 = 0.346, p < 0.05), FPG (r2 = 0.264, p < 0.05), insulin (r2 = 0.388, p < 0.05), HOMA-IR (r2 = 0.384, p < 0.05).

CONCLUSIONS

Serum FGF23 levels were higher in women with GDM compared with controls. The present findings suggest that FGF23 could be a useful marker of cardiovascular disease in GDM.

<em>Fibroblast</em> <em>growth</em> <em>factor</em> 21 (FGF21) is a major metabolic regulator that has been shown to be elevated in a number of metabolic disturbances including type 2 diabetes mellitus (T2DM) and the metabolic syndrome, but few studies about the relationship between serum FGF21 and the complications of diabetes have been done. Since the association between FGF21 and diabetic retinopathy is not clear, this study was conducted to investigate this relationship. In this cross-sectional study, 61 subjects (14 healthy controls, <em>22</em> diabetic patients without retinopathy, and 25 patients with diabetic retinopathy) were evaluated. All patients in the study were examined for the presence of diabetic retinopathy. Various clinical and biochemical parameters including FGF21 were evaluated and analyzed and compared between the study groups. Serum levels of FGF21 showed a significant difference between the three groups (P=0.003) but the difference between diabetic patients with and without retinopathy was not significant (P=0.1<em>22</em>). Regression model was used to evaluate the role of FGF21 in predicting diabetic retinopathy. In the multivariate logistic regression model after adjustment of systolic blood pressure and fasting blood glucose, the level of FGF21 was not associated with diabetic retinopathy. In the multivariate model, only fasting blood glucose was associated with diabetic retinopathy (P=0.009). According to the results of this study, serum levels of FGF21 in diabetic patients was higher than the control group but these raised levels could not predict the presence of diabetic retinopathy.

OBJECTIVE

To investigate the regulatory mechanism of MEN1 gene in radiation-induced lung fibrosis in mice and provide a new theoretical basis for the clinical treatment of radiation pulmonary fibrosis.

METHODS

First, 80 C57BL/6 mice aged 8 weeks and weighing 18-<em>22</em> g were selected, half of them were male and the other half were female. The mice were divided into control group and irradiation group (40 mice in each group) according to the method of the random number table. A radiation-induced lung fibrosis mouse model was established in which a single X-ray irradiation of 20 Gy was applied to the right lung in the irradiation group; H&E and Masson staining were used to verify whether the model was successful at 4, 8, 16 and 24 weeks after irradiation. The expression of MEN1, smooth muscle actin (α-SMA), Collagen-1 and transforming <em>growth</em> <em>factor</em> (TGF-β) in lung tissue were detected by Western blot and qPCR. Secondly, in the mouse embryonic <em>fibroblast</em> cell line (MEF) and mouse lung epithelial cell line (MLE-12), we constructed cell models of MEN1 knockout and interference separately with the irradiation of 10 Gy X-rays. The expression of α-SMA, Collagen-1, and TGF-β/Smads signaling pathway molecules was detected by qPCR. Finally, using the immunoprecipitation (IP) method, we can detect the interaction between Smad2 and the protein menin encoded by the MEN1 gene.

RESULTS

The results of the radiation pulmonary fibrosis model in mice showed that compared with the control group, the alveolar septum widens, the alveolar integrity decreases, the lung tissue slightly thickens, and a small amount of collagen deposits appear after 4-8 weeks in the model group. At twenty-fourth weeks, a large number of cells in the interstitial space of the lung tissue and a localized focal fibrosis area were observed. Further study found that radiation induced fibrogenic inflammatory cytokines TGF-β up-regulation, down-regulation of MEN1 gene expression, and then enhanced the expression of α-SMA and promotes the transformation of fibroblasts to myofibroblasts; At the same time, the expression of Collagen-1 was enhanced, which suggested that the extracellular matrix was overconcentrated and eventually promoted the formation of pulmonary fibrosis. In vitro, we found that knockout and interference of MEN1 gene can significantly enhance radiation-induced fibrosis, and up-regulate the expression of downstream molecules Smad2 and Smad3 of TGF-β signaling pathway, and down-regulate the expression of Smad7. Furthermore, it played an important role in regulating the process of radionuclide fibrosis.

CONCLUSIONS

MEN1 plays a key role in the formation of pulmonary fibrosis by regulating the secretion of TGF-β and the activation of TGF-β/Smads signaling pathway.

Purpose: To investigate the relationship of Fibroblast Growth Factor Receptor-4 (FGFR-4) expression with radiologic, pathologic, and clinical parameters in pituitary adenomas.

Methods: Among 307 patients who underwent pituitary surgery for a pituitary adenoma between 2000 and 2015, we included 161 patients (53 gonadotroph, 26 corticotroph, 25 null cell, 22 lactotroph, 13 somatotroph, 8 adenomas with unusual combination, 7 Pit-1 positive adenomas, and 7 lactosomatotroph) based on availability of pathology specimens. Patients' radiologic, pathologic, and clinical parameters were determined. FGFR-4 immunostaining was evaluated using a semi-quantitative histologic score (H-score).

Results: The mean follow-up period was 61 (IQR=32-84) months. The median H-scores for FGFR-4 were higher in patients without remission, those with residual lesion, and T2-hyperintense adenoma (p<0.05). Ki-67 level was higher in patients without remission compared to those in remission (p<0.05). The mean Ki-67 levels did not differ between patients with and without residual lesion or T2-hyperintense tumor (p>0.05). There was no significant difference (p>0.05) when the H-score and Ki-67 levels were assessed in terms of sex, sellar-dural invasion, Knosp and a grading system for superior, inferior, parasellar, anterior and posterior tumor extension Classification, tumor function or presence of poor subtype. Adenomas with Ki-67 expression ≥3% had higher FGFR4 expression levels than those with <3% expression (p=0.002). There was a weak positive correlation between H-score and Ki-67 (p=0.011; r=0.201).

Conclusions: Higher levels of FGFR-4 in pituitary adenomas could be use a marker for more aggressive tumor behavior.

A large proportion of patients with chronic kidney disease (CKD) are vitamin D deficient (plasma 25-hydroxyvitamin D (25(OH)D) < 25 or 30 nmol/L per UK and US population guidelines) and this contributes to the development of CKD-mineral bone disease (CKD-MBD). Gaps in the evidence-base for the management of vitamin D status in relation to CKD-MBD are hindering the formulation of comprehensive guidelines. We conducted a systemic review of <em>22</em> RCTs with different forms of vitamin D or analogues with CKD-MBD related outcomes and meta-analyses for parathyroid hormone (PTH). We provide a comprehensive overview of current guidelines for the management of vitamin D status for pre-dialysis CKD patients. Vitamin D supplementation had an inconsistent effect on PTH concentrations and meta-analysis showed non- significant reduction (P = 0.08) whereas calcifediol, calcitriol and paricalcitol consistently reduced PTH. An increase in <em>Fibroblast</em> <em>Growth</em> <em>Factor</em> 23 (FGF23) with analogue administration was found in all 3 studies reporting FGF23, but was unaltered in 4 studies with vitamin D or calcifediol. Few RCTS reported markers of bone metabolism and variations in the range of markers prevented direct comparisons. Guidelines for CKD stages G1-G3a follow general population recommendations. For the correction of deficiency general or CKD-specific patient guidelines provide recommendations. Calcitriol or analogues administration is restricted to stages G3b-G5 and depends on patient characteristics. In conclusion, the effect of vitamin D supplementation in CKD patients was inconsistent between studies. Calcifediol and analogues consistently suppressed PTH, but the increase in FGF23 with calcitriol analogues warrants caution.

Keywords: Chronic kidney disease; Guidelines; Systematic review; Vitamin D deficiency; Vitamin D supplementation.

Purpose: This study analyzes the potential of stromal platelet-derived growth factor receptor-beta (PDGFRb) expression as biomarker for radiotherapy (RT) benefit on ipsilateral breast events (IBE) in ductal carcinoma in situ (DCIS). Improved identification of DCIS patients refractory to adjuvant whole-breast RT is needed. Predictive biomarker studies in DCIS have focused on tumor cell features rather than the tumor-associated stroma, despite growing evidence of its influence on therapy efficiency.

Methods: Samples from the Swedish randomized radiotherapy DCIS trial (SweDCIS) were subjected to IHC analysis for stromal PDGFRb expression. IBE incidence at 10 years after breast-conserving surgery was the primary endpoint. Interactions between marker and treatment were analyzed.

Results: PDGFRb score was predictive for RT benefit with regard to IBE (P _interaction = 0.002 and P _interaction = 0.008 adjusted multivariably). Patients of the PDGFRb^low group had a strong benefit from RT regarding IBE risk [HR, 0.23; 95% confidence interval (CI), 0.12-0.45; P < 0.001] with an absolute risk reduction of 21% (cumulative risk 7% vs. 28%) at 10 years. No significant risk reduction by RT was observed for patients of the PDGFRb^high group (HR, 0.83; 0.51-1.34; P = 0.444; cumulative risk 22% vs. 25%). The RT response-predictive effect of stromal PDGFRb was equally strong in analyses for in situ and invasive IBE when analyzed separately (in situ IBE: P = 0.029; invasive IBE: P = 0.044).

Conclusions: Results suggest high stromal PDGFRb expression as a novel biomarker identifying DCIS patients who are refractory to standard whole-breast adjuvant RT. The data imply previously unrecognized fibroblast-mediated modulation of radiosensitivity of DCIS, which should be further explored from mechanistic and targeting perspectives.

Human acidic fibreblast <em>growth</em> <em>factor</em> (haFGF) was a kind of cell <em>growth</em> <em>factor</em> with wide bio-activity on cell from mesectoderm and neuro-ectoderm.In this paper, the effect of acetate concentration on the <em>growth</em> and expression of recombinant human acidic <em>fibroblast</em> <em>growth</em> <em>factor</em> mutant system E.coli BL21(DE3)/pET3C-haFGF was investigated. Four fed-batch modes: batch-fed, batch-DO static balance, DO static balance-glucose starvation, and pH-static state were investigated. The accumulation of acetate during the fermentation course was effectively inhibited. The OD600nm value was about <em>22</em>, after purification, the soluble rhaFGF yielded 450mg/L. During the fermentation, no special ways such as pure oxygen, pressure were adopted, thus the established process would be easily scaled up for industry purpose.

Idiopathic pulmonary fibrosis (IPF) is a chronic and progressive scarring disease with unknown etiology. The evidence of a pathogenic role for transforming <em>growth</em> <em>factor</em>-beta (TGF-β) in the development and progression of IPF is overwhelming. In the present study, we investigated the role of interleukin-<em>22</em> (IL-<em>22</em>) in the pathogenesis of IPF by regulating the TGF-β pathway. We measured parameters and tissue samples from a clinical cohort of IPF. IL-<em>22</em>R knock out (IL-<em>22</em>RA1^-/- ) and IL-<em>22</em> supplementation mouse models were used to determine if IL-<em>22</em> is protective in vivo. For the mechanistic study, we tested A549, primary mouse type II alveolar epithelial cell, human embryonic lung <em>fibroblast</em>, and primary <em>fibroblast</em> for their responses to IL-<em>22</em> and/or TGF-β1. In a clinical cohort, the expression level of IL-<em>22</em> in the peripheral blood and lung tissues of IPF patients was lower than healthy controls, and the lower IL-<em>22</em> expression was associated with poorer pulmonary function. IL-<em>22</em>R^-/- mice demonstrated exacerbated inflammation and fibrosis. Reciprocally, IL-<em>22</em> augmentation by intranasal instillation of recombinant IL-<em>22</em> repressed inflammation and fibrotic phenotype. In vitro, IL-<em>22</em> treatment repressed TGF-β1 induced gene markers representing epithelial-mesenchymal-transition and <em>fibroblast</em>-myo<em>fibroblast</em>-transition, likely via the inhibition of TGF-β receptor expression and subsequent Smad2/3 activation. IL-<em>22</em> appears to be protective against pulmonary fibrosis by inhibiting TGF-β1 signaling, and IL-<em>22</em> augmentation may be a promising approach to treat IPF.

<strong class="sub-title"> Keywords: </strong> IL-<em>22</em>; IPF; TGF-β; fibrosis.

<em>Growth</em> <em>factor</em> signaling in the brain is implicated in many neuropsychiatric disorders, including depression, autism, and epilepsy. <em>Fibroblast</em> <em>growth</em> <em>factor</em> <em>22</em> is a <em>growth</em> <em>factor</em> that regulates excitatory synapse development and neurogenesis in the brain. We have previously shown that adult mice in which <em>fibroblast</em> <em>growth</em> <em>factor</em> <em>22</em> is constitutively inactivated in all cells throughout life (<em>fibroblast</em> <em>growth</em> <em>factor</em> <em>22</em>-null mice) show anhedonia, a core feature of depression in humans, suggesting that <em>fibroblast</em> <em>growth</em> <em>factor</em> <em>22</em> signaling contributes to the regulation of affective behavior. Here we asked (1) whether inactivation of <em>fibroblast</em> <em>growth</em> <em>factor</em> <em>22</em> specifically in neurons is sufficient to induce anhedonia in mice and (2) whether <em>fibroblast</em> <em>growth</em> <em>factor</em> <em>22</em> signaling is important during development or in adults for the regulation of affective behavior. To address these questions, we performed the sucrose preference test, which is used as an indicator of anhedonia, with neuron-specific conditional <em>fibroblast</em> <em>growth</em> <em>factor</em> <em>22</em> knockout mice, in which <em>fibroblast</em> <em>growth</em> <em>factor</em> <em>22</em> is inactivated in neurons at birth (neonatal-<em>fibroblast</em> <em>growth</em> <em>factor</em> <em>22</em>-knockout mice) or in adults (adult-<em>fibroblast</em> <em>growth</em> <em>factor</em> <em>22</em>-knockout mice). We found that neonatal-<em>fibroblast</em> <em>growth</em> <em>factor</em> <em>22</em>-knockout mice show anhedonia (decreased preference for sucrose), while adult-<em>fibroblast</em> <em>growth</em> <em>factor</em> <em>22</em>-knockout mice do not. Therefore, neuronal <em>fibroblast</em> <em>growth</em> <em>factor</em> <em>22</em> signaling is critical during development, and not in adults, for the regulation of affective behavior. Our work also implies that defects in <em>growth</em> <em>factor</em>-dependent synapse development, neurogenesis, or both may underlie depression of a developmental origin.

To investigate the efficacy of Ciji Hua'ai Baosheng formula (CHBF) on microvessel density (MVD) and vascular endothelial <em>growth</em> <em>factor</em> (VEGF), kinase insert domain-containing receptor (KDR) and basic <em>fibroblast</em> <em>growth</em> <em>factor</em> (bFGF) expression in serum and tumor tissue of mice receiving chemotherapy for the treatment of H(<em>22</em>) hepatocellular carcinoma.

Sixty Kunming mice were injected subcutaneously with H(<em>22</em>) hepatoma carcinoma cell suspensions into the right anterior armpit. Seven days later, all transplanted tumor were formed and the mice were intraperitoneally injected 200 mg/kg Cytoxan (CTX) to establish the models of tumor-bearing mouse chemotherapy, then they were randomly divided into model group, continuing CTX chemotherapy group (CTX group), and three CHBF (117, 58.5 and 29.25 g/kg) groups. After ten days of treatments, histology was observed, contents of VEGF, KDR and bFGF in serum and tumor tissue were measured by enzyme-linked immunosorbent assay (ELISA), VEGF and bFGF protein expression and MVD tagged by CD34 were detected by immunohistochemistry.

MVD in CHBF (117, 58.5 g/kg) and CTX groups was significantly lower than that in model group (P < 0.01); expressions of VEGF, KDR and bFGF in serum and tumor tissue in CHBF (117 g/kg) group were less than those in model group (P < 0.05; P < 0.01); the expressions of MVD, VEGF and bFGF in tumor tissue of CHBF (117 g/kg) group were also less than those in CTX group (P < 0.05; P < 0.01).

CHBF can effectively reduce the expression of VEGF, KDR and bFGF in serum and tumor tissue, and decrease MVD and delay tumor progression.

Limb bud mesoderm of stage <em>22</em>-23 embryos was dissected into four pieces along the anteroposterior axis and dissociated cells of each region were separately cultured under various conditions. When the cells were cultured in medium containing 0.1% fetal calf serum (serum-poor medium) only a slight increase in cell number occurred in the cultures of all four regions. However, when the cells were cultured in medium containing 10% FCS, only cells of two central regions proliferated rapidly, and no <em>growth</em> promotion was observed in cells in the most anterior and posterior regions. Using the serum-poor medium, we examined the <em>growth</em>-promoting effects of cocultured limb bud fragments and of some <em>growth</em> <em>factors</em> on the cells of four regions. Anterior, distal, and proximal fragments promoted cell proliferation and their promotive effect on the cells of each region was equal. On the other hand, posterior fragments (containing ZPA) showed stronger promotive effects on preaxial cells than on postaxial cells. For comparison with the <em>growth</em>-promotive effect of the posterior fragment, <em>fibroblast</em> <em>growth</em> <em>factor</em> (FGF), epidermal <em>growth</em> <em>factor</em> (EGF), insulin, and retinoic acid were tested in cell culture. FGF showed position-dependent <em>growth</em> promotion, while EGF and insulin promoted <em>growth</em> in the cells of all four regions to a similar degree. Retinoic acid showed no effect on cell <em>growth</em> at low concentrations, and was rather toxic at high concentrations. These results suggest that the cells of the posterior region secrete an FGF-like <em>growth</em> <em>factor</em>(s), which controls normal limb development and experimental duplicate formation.

Purpose: Transamniotic stem cell therapy (TRASCET) with mesenchymal stem cells (MSCs) can induce spina bifida coverage with neoskin. We initiated a mechanistic analysis of this host response.

<strong class="sub-title"> Methods: </strong> Pregnant dams (n = 28) exposed to retinoic acid to induce fetal spina bifida were divided into an untreated group and 2 groups receiving intra-amniotic injections on gestational day 17 (E17; term = E21-<em>22</em>) of either amniotic fluid-derived MSCs (afMSCs; n = 105) or saline (n = 107). Gene expressions of multiple paracrine and cell clonality markers were quantified at term by RT-qPCR at the defect and fetal bone marrow. Defects were examined histologically for neoskin coverage. Comparisons were by Mann-Whitney U tests and logistic regression.

Results: Defect coverage was associated with significant downregulation of both epidermal growth factor (Egf; p = 0.031) and fibroblast growth factor-2 (Fgf-2; p = 0.042) expressions at the defect and with significant downregulation of transforming growth factor-beta-1 (Tgfb-1; p = 0.021) and CD45 (p = 0.028) expressions at the fetal bone marrow.

Conclusions: Coverage of experimental spina bifida is associated with local and bone marrow negative feedback of select paracrine factors, as well as increased relative mesenchymal stem cell activity in the bone marrow. Further analyses informed by these findings may lead to strategies of nonsurgical induction of prenatal coverage of spina bifida.

Keywords: Amniotic mesenchymal stem cell; Amniotic stem cell; Fetal stem cell; Myelomeningocele; Spina bifida; Transamniotic stem cell therapy.

Previous work aimed at understanding the gene regulatory networks (GRNs) governing caudal hindbrain formation identified morphogens such as Retinoic Acid (RA) and Fibroblast growth factors (FGFs), as well as transcription factors like hoxb1b, hoxb1a, hnf1ba, and valentino as being required for rhombomere (r) r4-r6 formation in zebrafish. Considering that the caudal hindbrain is relatively complex - for instance, unique sets of neurons are formed in each rhombomere segment - it is likely that additional essential genes remain to be identified and integrated into the caudal hindbrain GRN.

By taking advantage of gene expression data available in the Zebrafish Information Network (ZFIN), we identified 84 uncharacterized genes that are expressed in r4-r6. We selected a representative set of 22 genes and assayed their expression patterns in hoxb1b, hoxb1a, hnf1b, and valentino mutants with the goal of positioning them in the caudal hindbrain GRN. We also investigated the effects of RA and FGF on the expression of this gene set. To examine whether these genes are necessary for r4-r6 development, we analyzed germline mutants for six of the genes (gas6, gbx1, sall4, eglf6, celf2, and greb1l) for defects in hindbrain development.

Our results reveal that r4 gene expression is unaffected by the individual loss of hoxb1b, hoxb1a or RA, but is under the combinatorial regulation of RA together with hoxb1b. In contrast, r5/r6 gene expression is dependent on RA, FGF, hnf1ba and valentino - as individual loss of these factors abolishes r5/r6 gene expression. Our analysis of six mutant lines did not reveal rhombomere or neuronal defects, but transcriptome analysis of one line (gas6 mutant) identified expression changes for genes involved in several developmental processes - suggesting that these genes may have subtle roles in hindbrain development.

We conclude that r4-r6 formation is relatively robust, such that very few genes are absolutely required for this process. However, there are mechanistic differences in r4 versus r5/r6, such that no single factor is required for r4 development while several genes are individually required for r5/r6 formation.

<AbstractText>To demonstrate that superficial high-dose-rate (HDR) brachytherapy by means of Leipzig applicators or moulds with catheters is an adjuvant treatment with impact on local control and low toxicity.</AbstractText><AbstractText>Keloid scars occur in 5-15 % of cases, secondary to an uncontrolled proliferation of <em>fibroblasts</em> and reduction in the inhibition of <em>growth</em> <em>factors</em>.</AbstractText><AbstractText>Retrospective, longitudinal and descriptive study in patients with keloid scars who were treated with superficial HDR brachytherapy in the General Hospital of Mexico between November 2009 and December 2013.</AbstractText><AbstractText>Eighty patients were evaluated, and the mean follow-up was <em>22</em>.18 months (range 8-48). The anatomic site treated was the ear in 72 patients (90.0 %), anterior thorax in 5 patients, retroauricular region in 2, and abdomen region in 1 patient. The application was performed 24 h after surgery; the dose for 79 patients (99 %) was 1500 cGy/3 fractions, and 1 received 500 cGy in 1 fraction. Adequate healing occurred in 76 patients (95 %), and the local failure was 5 % (95 % CI). Acute toxicity occurred in 15 % (12 patients) with grade 1 radioepithelitis. Chronic toxicity occurred in <em>22</em> patients (27.5 %) with grade 1 hypopigmentation and 18 patients (<em>22</em>.5 %) with grade 1 fibrosis. The cosmetic result was good in 72 patients (90 %). During follow-up, 2 patients presented recurrence, and 2 patients persisted.</AbstractText><AbstractText>Treatment with superficial brachytherapy in keloid scars using a mould with catheters or a Leipzig applicator is a therapeutic option that results in 95 % local control and low toxicity.</AbstractText>

In order to investigate the epitope of basic <em>fibroblast</em> <em>growth</em> <em>factor</em> (bFGF) and its immunogenicity, the epitopes of bFGF were screened from the phage display library with monoclonal antibody GF<em>22</em>, which can neutralize the bio-activity of bFGF. By three rounds of screening, the positive phage clones with bFGF epitopes were selected, which can effectively block the bFGF to bind with GF<em>22</em>. Sequence analysis showed that the epitopes shared a highly conservative sequence (Leu-Pro-Pro/Leu-Gly-His-Phe/Ile-Lys). The sequence of PPGHFK was located at <em>22</em>-27 of the bFGF. The specific immuno-response of mouse could be highly induced by phage clones with the epitopes. And the anti-bFGF activity induced by LPGHFK was 3 times higher than the original sequence, which showed that the mimetic peptide LPLGHIK might be used as a tumor vaccine in the prevention and treatment of tumor.

OBJECTIVE

To look into the character of the expression of collagen type II and transforming growth factor beta1 (TGF-beta1), basic fibroblast growth factor (bFGF) in the apical articular process cartilages of adolescent idiopathic scoliosis (AIS) and congenital scoliosis (CS) patients.

METHODS

The articular processes of 22 AIS and 18 CS were collected. The techniques of HE staining, immunohistochemistry and in situ hybridization were adopted in this research. By comparing the apical processes with the end processes, the convex processes with the concave processes, the AIS processes with CS processes, the pathological changes of the articular process cartilages of these patients and the distribution of collagen type II and TGF-beta1, bFGF in them were studied. The images of immunohistochemistry and in situ hybridization were input into the image analysis system and were analyzed semi-quantitatively. The SAS software (8.01) was adopted, and P < 0.05 was defined as the significant level.

RESULTS

The expression of collagen type II and TGF-beta1, bFGF in AIS was similar to CS: the concave sides of apexes were higher than the convex sides. The comparisons had statistical significance. There was no statistical significance between upper and lower end vertebrae in convex and concave sides, between convex and concave sides in upper and lower end vertebrae. The apical vertebrae were significantly higher than the ipsilateral sides of upper or lower end vertebrae for collagen type II. There was no statistical difference of the expression at the concave, convex, upper, lower end vertebrae between AIS and CS.

CONCLUSIONS

The cartilages of the apical processes show some signs of regression and hypoplasia in scoliosis. The concave side is more severe than the convex side. Increase of collagen type II and TGF-beta1, bFGF in the concave sides of apical processes in scoliosis may be the results of reconstruction of extracellular matrix and the compensation reactions which are caused by abnormal biomechanical forces such as compressive stresses. Compressive stress on the concave sides has more influences on the expression of collagen type II than tensile stress on the convex sides.

Insulin-like <em>growth</em> <em>factor</em>-II (IGF-II) and lysosomal enzymes bearing the mannose 6-phosphate (Man6P) recognition marker, bind to two distinct binding sites of the IGF-II/M6P receptor. The two classes of ligands reciprocally modulate the binding of the other class of ligand to the receptor [Kiess, W., Thomas, C. L., Greenstein, L., Lee, L., Sklar, M. M., Rechler, M. M., Sahagian, G. G. & Nissley, S. P. (1989) J. Biol. Chem. 264, 4710-4714]. We asked whether or not overexpression of pro-IGF-II by cells in culture leads to missorting of lysosomal enzymes. Human embryonal kidney <em>fibroblasts</em> were transfected with the full-length human IGF-II cDNA or a control cDNA. Solution hybridization/RNase protection experiments using a human IGF-II riboprobe showed that two transfectants expressed large quantities of IGF-II mRNA, whereas the non-transfected cells did not. The analysis of conditioned media revealed that these cells secrete approximately 0.15 micrograms and 1.0 micrograms immunoreactive IGF-II/ml and <em>22</em> x 10(6) cells and 24 x 10(6) cells within 24 hours. Immunoreactive IGF-II was shown by Western blotting to represent 17-kDa pro-IGF-II. The amount of the lysosomal enzyme, beta-hexosaminidase, was approximately twofold increased in the conditioned media from pro-IGF-II overexpressing cells compared with control media, as shown by Western-blot analysis and immunoprecipitation of media extracts of metabolically labeled cells. The synthesis rate of beta-hexosaminidase was not affected by pro-IGF-II overexpression. In addition, the basal amount of another newly synthesized lysosomal enzyme, the cathepsin D precursor, was also twofold higher in pro-IGF-II overexpressing cells than in control cells. In contrast, the surface binding and cellular uptake rate of a Man6P-containing neoglycoprotein did not differ between the cell lines. The results indicate that the overexpression of pro-IGF-II doubles the secretion and/or reduces the re-uptake of beta-hexosaminidase and cathepsin D to approximately 20% of the total synthesized enzymes in human embryonal kidney <em>fibroblasts</em> compared to control cells. We hypothesize that, in cells synthesizing high amounts of pro-IGF-II, the <em>growth</em> <em>factor</em> may modulate the targeting of a portion of lysosomal enzymes, mainly by partially enhancing the secretion of newly synthesized enzymes and, in addition, possibly by affecting the re-uptake mechanism.

Phytase hydrolyzes phytate rendering phosphorus available for intestinal absorption, while systemic neutralization of <em>fibroblast</em> <em>growth</em> <em>factor</em> 23 (FGF-23), using anti-FGF-23 antibody, has been shown to increase phosphate retention. Hence, neutralization of FGF-23 should be additive with phytase in reducing dietary non-phytate phosphorus (nPP) needs in chickens fed plant-based diets rich in phytic acid. This study was designed to test the additive effects of maternally derived anti-FGF-23 antibody and dietary phytase on the performance of chicks fed a low nPP diet from one to 14 d. Single Comb White Leghorn laying hens were vaccinated with either an adjuvant control or a synthetic FGF-23 peptide (GMNPPPYS). Chicks from vaccinated hens with control or anti-FGF-23 maternal antibodies were fed either a diet containing 0.2% nPP and 0.9% calcium with or without 500 unit phytase per kg of diet (2 × 2 <em>factor</em>ial with main effects of antibody type and phytase addition, n = 15 pens of chicks/treatment). A significant interaction between dietary phytase and maternally derived anti-FGF-23 antibody on <em>growth</em> and feed efficiency was observed (P ≤ 0.05), in which chicks receiving either phytase or maternally derived anti-FGF-23 antibody had improved body weight gain (21 or 15%, respectively) and feed efficiency (16 or 18%, respectively) as compared to chicks with control antibody and not fed phytase. Both phytase and maternally derived anti-FGF-23 antibody independently increased (P ≤ 0.05) plasma phosphate (11 and 11%, respectively) and percent tibiotarsus ash (13 and 11%, respectively). Significant main effects and the lack of an interaction supported an additive effect of phytase and anti-FGF-23 antibody on plasma phosphate and percent tibiotarsus ash. Feeding phytase to chicks fed 0.2% nPP increased plasma FGF-23 levels by <em>22</em>% (P ≤ 0.05); however, no effects of anti-FGF-23 antibody on plasma FGF-23 levels were observed. In conclusion, dietary phytase and presence of anti-FGF-23 antibody have an additive effect on plasma phosphate and tibiotarsus ash in chicks fed low nPP diets. Data support that phytase and anti-FGF-23 antibody increase phosphate utilization by different mechanisms.

Platelet-derived <em>growth</em> <em>factor</em> (PDGF) and basic <em>fibroblast</em> <em>growth</em> <em>factor</em> (bFGF) have been implicated in myointimal proliferative arteriopathy, a lesion seen in monocrotaline-induced pulmonary hypertension (MIPH). The purpose of this study was to examine the expression of PDGF and bFGF in the lungs of rats given monocrotaline and to examine the effects of amrinone on the hearts and lungs of these rats. Twenty-four 26-day-old rats were randomized to receive either monocrotaline (approximately 3.6 mg/kg/d) or no monocrotaline and concomitantly to receive either amrinone (100 mg/kg/d) or no amrinone for 21 days. Lungs were examined for immunohistochemical evidence of PDGF and bFGF, and hearts were examined for effects of pulmonary hypertension and amrinone. Immunohistochemical staining of lungs showed no evidence of PDGF except in bronchioles. bFGF staining was similar between groups (no monocrotaline 25%, monocrotaline 27%, monocrotaline and amrinone <em>22</em>%), and the staining was confined to the arterial walls. Rats given monocrotaline showed significantly greater right ventricular (RV) weight (0.13 +/- 0.02 g versus 0.23 +/- 0.04 g [mean +/- SD], P < 0.001), right ventricular/left ventricular (RV/LV) weight ratio (0.29 +/- 0.06 versus 0.59 +/- 0.1, P < 0.001), and lung/body weight ratio (0.006 +/- 0.001 versus 0.01 +/- 0.003, P < 0.05) than controls. Rats given monocrotaline and amrinone were not significantly different from rats given only monocrotaline with regard to RV weight, RV/LV weight ratio, or lung/body weight ratio. We conclude that the vasculopathy seen in MIPH is not associated with the presence of PDGF or bFGF, suggesting that other <em>growth</em> <em>factors</em> may mediate this process. The course of MIPH is not altered by amrinone.

Cerebroside sulfotransferase (EC 2.8.2.11, CST) specific activity has been determined in oligodendrocyte (OL)-enriched glial cell cultures from newborn rat brain grown in serum supplemented medium. This activity is detectable at 5 days in vitro (DIV) and reaches its maximum value at 12 DIV. This period corresponds to that of oligodendrocyte precursor proliferation in these cultures. The activity decreases thereafter and remains nearly constant after 24 DIV. The developmental curve of CST activity is parallel in pure oligodendrocyte subcultures but twice higher than in primary cultures. These data confirm that CST is highly enriched in OL. Basic <em>fibroblast</em> <em>growth</em> <em>factor</em> (bFGF) (15 ng/ml) and platelet derived <em>growth</em> <em>factor</em> (PDGF) (0.75 U/ml) both enhance CST activity by 90% and 72%, respectively. This increase is in the same range than that of DNA content in treated cultures, whereas protein increase is smaller (50% and <em>22</em>%, respectively). In contrast, transforming <em>growth</em> <em>factor</em> beta 1 (TGF beta 1, 0.5 and 5 ng/ml) does not significantly enhance CST activity nor DNA content of OL cultures. Insulin at high concentrations (5 micrograms/ml) also enhances CST activity but has no effect at physiological concentrations (20 ng/ml). These results show that CST activity can be controlled by <em>growth</em> <em>factors</em>. They suggest that CST activity is more closely related to OL and OL precursor proliferation than to myelination itself since its maximal activity preceeds myelination in vitro.

OBJECTIVE

We present perinatal imaging findings and molecular genetic analysis of thanatophoric dysplasia type I (TD1) in a fetus.

METHODS

A 28-year-old woman was referred for genetic counseling at <em>22</em> weeks of gestation because of abnormal prenatal ultrasound findings. Level II ultrasound examination revealed a narrow chest, shortened and curved long limbs, protrusion of the abdomen, and macrocephaly. A tentative diagnosis of TD1 was made. After genetic counseling, the pregnancy was terminated and a malformed fetus was delivered. Postnatal radiography findings were consistent with the diagnosis of TD1, with additional findings of short ribs, platyspondyly, and horizontal acetabular roofs. Molecular genetic analysis using umbilical cord tissue revealed a heterozygous mutation of c.2419T>G (p.Ter807Gly) (X807G) in the <em>fibroblast</em> <em>growth</em> <em>factor</em> receptor 3 gene (FGFR3).

CONCLUSIONS

A second-trimester fetus with a heterozygous c.2419T>G mutation in FGFR3 may present characteristic ultrasound and X-ray findings of TD1.

Acute respiratory distress syndrome (ARDS) has high rates of mortality and multisystem morbidity. Pre-clinical data suggest that <em>fibroblast</em> <em>growth</em> <em>factor</em> 23 (FGF23) may contribute to pulmonary pathology, and FGF23 is associated with mortality and morbidity, including acute kidney injury (AKI), in non-ARDS cohorts. Here, we assess whether FGF23 is associated with AKI and/or mortality in a cohort of 161 pediatric ARDS patients. Plasma total (intact + C-terminal) FGF23 and intact FGF23 concentrations were measured within 24 hours of ARDS diagnosis (Day 1), and associations with Day 3 AKI and 60-day mortality were evaluated. 35 patients (<em>22</em>%) developed AKI by 3 days post-ARDS diagnosis, and 25 (16%) died by 60 days post-ARDS diagnosis. In unadjusted models, higher Day 1 total FGF23 was associated with Day 3 AKI (odds ratio (OR) 2.<em>22</em> [95% confidence interval (CI) 1.62, 3.03], p<0.001), but Day 1 intact FGF23 was not. In a model adjusted for demographics and disease severity, total FGF23 remained associated with AKI (OR 1.52 [95% CI 1.02, 2.26], p = 0.039). In unadjusted models, both higher Day 1 total and intact FGF23 were associated with 60-day mortality (OR 1.43 [95% CI 1.07, 1.91], p = 0.014; and OR 1.44 [95% CI 1.02, 2.05], p = 0.039, respectively). In the adjusted model, only total FGF23 remained associated with 60-day mortality (OR 1.62 [95% CI 1.07, 2.45], p = 0.023). In a subgroup analysis of patients with Day 1 plasma IL-6 concentrations available, inflammation partially mediated the association between total FGF23 and AKI. Our data suggest both inflammation-dependent and inflammation-independent associations between total FGF23 and clinical outcomes in pediatric ARDS patients.

Achondroplasia is a genetic mutation of <em>fibroblast</em> <em>growth</em> <em>factor</em> receptor resulting in impaired <em>growth</em> plate development in long bones due to lower collagen turnover. Despite the characteristic shorter stature and lower strength in Achondroplasic groups, little is known of the tendon mechanical properties under loading. The aim of this study was therefore to conduct a between measure design of patella tendon (PT) mechanical properties (stress, strain, stiffness and Young's Modulus) in 10 men with Achondroplasia (<em>22</em> ± 3 years) and 17 male controls (<em>22</em> ± 2 years). PT mechanical properties were measured during isometric maximal voluntary contraction (iMVC) of the knee extensors using ultrasonography. The Achondroplasic group produced 54% less stress at iMVC than controls (29.4 ± 8.0 v 64.5 ± 14.0 MPa, P < 0.001, d = 3.12). Maximal excursion of the Achondroplasic PT was <em>22</em>% less than controls at iMVC (7.4 ± 2.1 v 5.5 ± 1.7 mm, P < 0.001, d = 0.99), but there was no difference in strain between groups (13 ± 4 v 13 ± 3%, P>> 0.05). Achondroplasic PT were 47% less stiff (748 ± 93 v 1418 ± 101 N·mm-1, P < 0.001, d = 6.89) and had a 51% lower Young's modulus (0.39 ± 0.09 v 0.77 ± 0.14 GPa, P < 0.001, d = 3.46) than controls at iMVC. Achondroplasic PT are indeed more compliant than controls which may contribute to lower relative force production. The causes of higher Achondroplasic PT compliance are unclear but are likely due to the collagen related genetic mutation which causes Achondroplasia.

OBJECTIVE

To study the gene mutation of Chinese patients with achondroplasia(ACH) and to set up a simple and rapid molecular diagnostic method to differentiate ACH from other similar genetic dwarfism.

METHODS

The specific fragment of fibroblast growth factor receptor 3(FGFR3) transmembrane domain was amplified from dried blood spots of 21 patients with ACH and 6 suspicious patients with ACH by polymerase chain reaction, then mutation was screened and detected by restrictive enzyme analysis, single strand conformation polymorphism(SSCP) and denaturing gradient gel electrophoresis(DGGE).

RESULTS

One out of 6 suspicious cases was ACH and 5 were pseudoachondroplasia(PSACH). Twenty-one out of 22 patients with ACH bore a G to A transition at nucleotide 1138 and 1 bore a G to C transversion at this same position.

CONCLUSIONS

The nucleotide 1138 of FGFR3 gene is also the hotspot of mutation in Chinese patients with ACH. A simple and rapid molecular diagnostic method has been set up to differentiate ACH from other similar genetic dwarfism.