Pre- and Postoperative Magnetic Resonance Imaging Comparative Studies of Pituitary Tissues Following Transsphenoidal Resection of Pituitary Macroadenomas.

BACKGROUND: Magnetic resonance imaging (MRI) has become the most important radiological procedure for diagnosing and following pituitary tumors. But previous MRI studies on pituitary adenomas are mainly focused on the posterior pituitary. Few research has been done on residual normal pituitary tissue before and after transsphenoidal surgery. This retrospective cohort study investigates the pre- and postoperative magnetic resonance imaging characteristics of normal pituitary tissues regarding transsphenoidal resection of pituitary macroadenomas. METHODS: Pre- and postoperative magnetic resonance imaging scanning of 112 consecutive pituitary macroadenoma patients who underwent tumor resection via transsphenoidal approach was performed, and their medical records were studied. RESULTS: On preoperative MRI, 66 cases of pituitary stalks were identifiable, 9 of them were roughly in the middle, and 57 cases showed left or right deviation, with the angle between pituitary stalks and the sagittal plane was 5.32°-64.05° (average 21.65°). Among the 57 patients with preoperative pituitary stalk deviation, 55 of the pituitary stalk deviations improved in 1 week after surgery, and 30 cases were almost in the middle in 4-6 months after operation, with the other cases get better in varying degrees. The diameter of pituitary stalk was 1.08-3.89 mm (mean 2.36 mm) in pre-operation, and 1.29-3.43 mm (mean 2.30 mm) in 4-6 months after operation. The length of pituitary stalk was 1.41-11.74 mm (mean 6.12 mm) preoperatively, 3.61-11.63 mm (mean 6.93 mm) early postoperatively, and 5.37-17.57 mm (mean 8.83 mm) in 4-6 months after operation. Pituitary stalk was thickened or compressed on preoperative MR images, and gradually recovered to normal during postoperative period. It tended to be in the middle position and its length increased gradually until 4-6 months after operation. On preoperative MRI, 69 out of 112 patients showed residual pituitary tissues (RPT)(+) on enhanced MRI. RPT were likely located above the adenomas in somatotroph adenoma patients. Morphological restitution of postoperative normal pituitary tissues was better in lateral displacement than in superior or superolateral patterns on preoperative magnetic resonance imaging. Postoperative normal pituitary tissues usually subsided directly in superior displacement pattern on preoperative MRI, while were likely to be confined in the lateral side in lateral and superolateral displacement patients. Postoperative morphologic remodeling grade of RPT was positively correlated with the maximum diameter of pituitary adenoma (p = 0.000), but not with age. CONCLUSIONS: The larger the tumor diameter, the worse the pituitary morphological recovery after tumor resection. Relative locations of normal pituitary and adenoma tissues may be related to adenoma type and may affect postoperative reconstruction of residual normal pituitary tissues. These findings enable surgeons to distinguish pituitary tissue from residual or recurring tumor tissue on postoperative magnetic resonance imaging.

The roles of SMYD4 in epigenetic regulation of cardiac development in zebrafish.

SMYD4 belongs to a family of lysine methyltransferases. We analyzed the role of smyd4 in zebrafish development by generating a smyd4 mutant zebrafish line (smyd4L544Efs*1) using the CRISPR/Cas9 technology. The maternal and zygotic smyd4L544Efs*1 mutants demonstrated severe cardiac malformations, including defects in left-right patterning and looping and hypoplastic ventricles, suggesting that smyd4 was critical for heart development. Importantly, we identified two rare SMYD4 genetic variants in a 208-patient cohort with congenital heart defects. Both biochemical and functional analyses indicated that SMYD4(G345D) was pathogenic. Our data suggested that smyd4 functions as a histone methyltransferase and, by interacting with HDAC1, also serves as a potential modulator for histone acetylation. Transcriptome and bioinformatics analyses of smyd4L544Efs*1 and wild-type developing hearts suggested that smyd4 is a key epigenetic regulator involved in regulating endoplasmic reticulum-mediated protein processing and several important metabolic pathways in developing zebrafish hearts.

BMP10 preserves cardiac function through its dual activation of SMAD-mediated and STAT3-mediated pathways.

Bone morphogenetic protein 10 (BMP10) is a cardiac peptide growth factor belonging to the transforming growth factor ß superfamily that critically controls cardiovascular development, growth, and maturation. It has been shown that BMP10 elicits its intracellular signaling through a receptor complex of activin receptor-like kinase 1 with morphogenetic protein receptor type II or activin receptor type 2A. Previously, we generated and characterized a transgenic mouse line expressing BMP10 from the α-myosin heavy chain gene promoter and found that these mice have normal cardiac hypertrophic responses to both physiological and pathological stimuli. In this study, we report that these transgenic mice exhibit significantly reduced levels of cardiomyocyte apoptosis and cardiac fibrosis in response to a prolonged administration of the ß-adrenoreceptor agonist isoproterenol. We further confirmed this cardioprotective function with a newly generated conditional Bmp10 transgenic mouse line, in which Bmp10 was activated in adult hearts by tamoxifen. Moreover, the intraperitoneal administration of recombinant human BMP10 was found to effectively protect hearts from injury, suggesting potential therapeutic utility of using BMP10 to prevent heart failure. Gene profiling and biochemical analyses indicated that BMP10 activates the SMAD-mediated canonical pathway and, unexpectedly, also the signal transducer and activator of transcription 3 (STAT3)-mediated signaling pathway both in vivo and in vitro Additional findings further supported the notion that BMP10's cardioprotective function likely is due to its dual activation of SMAD- and STAT3-regulated signaling pathways, promoting cardiomyocyte survival and suppressing cardiac fibrosis.

Two novel ANK1 loss-of-function mutations in Chinese families with hereditary spherocytosis.

Hereditary spherocytosis (HS) is the most common inherited haemolytic anaemia disorder. ANK1 mutations account for most HS cases, but pathogenicity analysis and functional research have not been widely performed for these mutations. In this study, in order to confirm diagnosis, gene mutation was screened in two unrelated Chinese families with HS by a next-generation sequencing (NGS) panel and then confirmed by Sanger sequencing. Two novel heterozygous mutations (c.C841T, p.R281X and c.T290G, p.L97R) of the ANK1 gene were identified in the two families respectively. Then, the pathogenicity of the two new mutations and two previously reported ANK1 mutations (c.C648G, p.Y216X and c.G424T, p.E142X) were studied by in vitro experiments. The four mutations increased the osmotic fragility of cells, reduced the stabilities of ANK1 proteins and prevented the protein from localizing to the plasma membrane and interacting with SPTB and SLC4A1. We classified these four mutations into disease-causing mutations for HS. Thus, conducting the same mutation test and providing genetic counselling for the two families were meaningful and significant. Moreover, the identification of two novel mutations enriches the ANK1 mutation database, especially in China.

Protein phosphatase 5 and the tumor suppressor p53 down-regulate each other's activities in mice.

Protein phosphatase 5 (PP5), a serine/threonine phosphatase, has a wide range of biological functions and exhibits elevated expression in tumor cells. We previously reported that pp5-deficient mice have altered ataxia-telangiectasia mutated (ATM)-mediated signaling and function. However, this regulation was likely indirect, as ATM is not a known PP5 substrate. In the current study, we found that pp5-deficient mice are hypersensitive to genotoxic stress. This hypersensitivity was associated with the marked up-regulation of the tumor suppressor tumor protein p53 and its downstream targets cyclin-dependent kinase inhibitor 1A (p21), MDM2 proto-oncogene (MDM2), and phosphatase and tensin homolog (PTEN) in pp5-deficient tissues and cells. These observations suggested that PP5 plays a role in regulating p53 stability and function. Experiments conducted with p53+/-pp5+/- or p53+/-pp5-/- mice revealed that complete loss of PP5 reduces tumorigenesis in the p53+/- mice. Biochemical analyses further revealed that PP5 directly interacts with and dephosphorylates p53 at multiple serine/threonine residues, resulting in inhibition of p53-mediated transcriptional activity. Interestingly, PP5 expression was significantly up-regulated in p53-deficient cells, and further analysis of pp5 promoter activity revealed that p53 strongly represses PP5 transcription. Our results suggest a reciprocal regulatory interplay between PP5 and p53, providing an important feedback mechanism for the cellular response to genotoxic stress.

Atrial fibrillation and electrophysiology in transgenic mice with cardiac-restricted overexpression of FKBP12.

Cardiomyocyte-restricted overexpression of FK506-binding protein 12 transgenic (αMyHC-FKBP12) mice develop spontaneous atrial fibrillation (AF). The aim of the present study is to explore the mechanisms underlying the occurrence of AF in αMyHC-FKBP12 mice. Spontaneous AF was documented by telemetry in vivo and Langendorff-perfused hearts of αMyHC-FKBP12 and littermate control mice in vitro. Atrial conduction velocity was evaluated by optical mapping. The patch-clamp technique was applied to determine the potentially altered electrophysiology in atrial myocytes. Channel protein expression levels were evaluated by Western blot analyses. Spontaneous AF was recorded in four of seven αMyHC-FKBP12 mice but in none of eight nontransgenic (NTG) controls. Atrial conduction velocity was significantly reduced in αMyHC-FKBP12 hearts compared with NTG hearts. Interestingly, the mean action potential duration at 50% but not 90% was significantly prolonged in αMyHC-FKBP12 atrial myocytes compared with their NTG counterparts. Consistent with decreased conduction velocity, average peak Na+ current ( INa) density was dramatically reduced and the INa inactivation curve was shifted by approximately +7 mV in αMyHC-FKBP12 atrial myocytes, whereas the activation and recovery curves were unaltered. The Nav1.5 expression level was significantly reduced in αMyHC-FKBP12 atria. Furthermore, we found increases in atrial Cav1.2 protein levels and peak L-type Ca2+ current density and increased levels of fibrosis in αMyHC-FKBP12 atria. In summary, cardiomyocyte-restricted overexpression of FKBP12 reduces the atrial Nav1.5 expression level and mean peak INa, which is associated with increased peak L-type Ca2+ current and interstitial fibrosis in atria. The combined electrophysiological and structural changes facilitated the development of local conduction block and altered action potential duration and spontaneous AF. NEW & NOTEWORTHY This study addresses a long-standing riddle regarding the role of FK506-binding protein 12 in cardiac physiology. The work provides further evidence that FK506-binding protein 12 is a critical component for regulating voltage-gated sodium current and in so doing has an important role in arrhythmogenic physiology, such as atrial fibrillation.

Evaluation of a Novel General Pituitary Hormone Score to Evaluate the Function of the Residual Anterior Pituitary (Adenohypophysis) in Patients Following Surgery for Pituitary Adenoma.

BACKGROUND The aim was to develop and assess a general pituitary hormone score to evaluate the function of the anterior pituitary (adenohypophysis) in patients following resection of pituitary adenomas. MATERIAL AND METHODS Sixty-six patients with pituitary null cell macroadenoma (1-3 cm diameter) (N=38) and pituitary null cell giant adenoma (≥3 cm diameter) (N=28) had preoperative and postoperative data including magnetic resonance imaging (MRI) and measurement of six pituitary hormones levels, adrenocorticotropic hormone (ACTH), growth hormone (GH), thyroid-stimulating hormone (TSH), prolactin (PRL), follicle-stimulating hormone (FSH), and luteinizing hormone (LH). The postoperative general pituitary hormone score, for 57 patients who underwent subtotal resection (>60%) and nine patients who underwent partial resection (≤60%), was 1-5 for each hormone level (score range, 6-30). RESULTS ACTH, GH, TSH, PRL, FSH, and LH levels in 38 patients with pituitary null cell macroadenoma were not statistically different from the 28 patients with pituitary null cell giant adenoma; the general pituitary hormone score in the former group was significantly increased compared with the latter group (P<0.05). ACTH, GH, TSH, PRL, FSH, and LH levels in the 57 patients with subtotal tumor resection were not significantly different from the nine patients with partial tumor resection; the general pituitary hormone score in the former group was significantly reduced compared with the latter group (P<0.05). CONCLUSIONS A general pituitary hormone score was developed that might be relevant to the evaluation of pituitary function following surgical resection of pituitary null cell macroadenoma and giant adenoma.

The Relationship Between Posterior Pituitary Bright Spot on Magnetic Resonance Imaging (MRI) and Postoperative Diabetes Insipidus for Pituitary Adenoma Patients.

BACKGROUND This study is to investigate the predictive value of posterior pituitary bright spot (PPBS) on magnetic resonance imaging (MRI) for postoperative diabetes insipidus (DI) in patients with pituitary adenoma. MATERIAL AND METHODS This was a retrospective study. In total, 65 patients with pituitary adenoma who underwent transsphenoidal surgery were enrolled. Before surgery, all patients had MRI examinations. The length of pituitary stalk and position of PPBS in T1WI sagittal and coronal sections were analyzed. The volume and height of the tumor was calculated in enhanced T1WI. Urine volume was monitored to analyze the clinical factors contributing to DI. RESULTS Among the 65 cases of pituitary adenoma, there were 54 cases of positive PPBS and 11 cases of negative PPBS. There were 32 cases of transient DI, and among these, 22 cases were positive PPBS and 10 cases were negative PPBS. However, there were 33 cases without DI, and among these, 32 cases were positive PPBS and one case was negative PPBS. The negative PPBS was significantly higher in cases with DI, compared with positive PPBS (P<0.05). Logistic regression showed that preoperative negative PPBS was an important predictor for postoperative DI (P<0.05). CONCLUSIONS Postoperative DI should be considered when there is negative preoperative PPBS on MRI. Also, severe pituitary stalk compression indicates higher risk of postoperative DI.

Multiple gene variations contributed to congenital heart disease via GATA family transcriptional regulation.

BACKGROUND: Congenital heart disease (CHD) is a common birth defect, and most cases occur sporadically. Mutations in key genes that are responsible for cardiac development could contribute to CHD. To date, the genetic causes of CHD remain largely unknown. METHODS: In this study, twenty-nine candidate genes in CHD were sequenced in 106 patients with Tetralogy of Fallot (TOF) using target exome sequencing (TES). The co-immunoprecipitation (CO-IP) and luciferase reporter gene assays were performed in HEK293T cells, and wild-type and mutant mRNA of ZFPM2 were microinjected into zebrafish embryos. RESULTS: Rare variants in key cardiac transcriptional factors and JAG1 were identified in the patients. Four patients carried multiple gene variants. The novel E1148K variant was located at the eighth Zinc-finger domain of FOG2 protein. The CO-IP assays in the HEK293T cells revealed that the variant significantly damaged the interaction between ZFPM2/FOG2 and GATA4. The luciferase reporter gene assays revealed that the E1148K mutant ZFPM2 protein displayed a significantly greater inhibition of the transcriptional activation of GATA4 than the wild-type protein. The wild-type mRNA and the E1148K mutant mRNA of ZFPM2 were injected into zebrafish embryos. At 48 hpf, in the mutant mRNA injection group, the number of embryos with an abnormal cardiac chamber structure and a loss of left-right asymmetry was increased. By 72 hpf, the defects in the chamber and left-right asymmetry became obvious. CONCLUSIONS: We performed TES in sporadic TOF patients and identified rare variants in candidate genes in CHD. We first validated the E1148 K variant in ZFPM2, which is likely involved in the pathogenesis of CHD via GATA4. Moreover, our results suggest that TES could be a useful tool for discovering sequence variants in CHD patients.

Efficacy of sellar opening in the pituitary adenoma resection of transsphenoidal surgery influences the degree of tumor resection.

BACKGROUND: Endonasal transsphenoidal microsurgery is often adopted in the resection of pituitary adenoma, and has showed satisfactory treatment and minor injuries. It is important to accurately localize sellar floor and properly incise the bone and dura matter. METHODS: Fifty-one patients with pituitary adenoma undergoing endonasal transsphenoidal microsurgery were included in the present study. To identify the scope of sellar floor opening, CT scan of the paranasal sinus and MRI scan of the pituitary gland were performed for each subject. Intraoperatively, internal carotid artery injury, leakage of cerebrospinal fluid, and tumor texture were recorded, and postoperative complications and residual tumors were identified. RESULT: The relative size of sellar floor opening significantly differed among the pituitary micro-, macro- and giant adenoma groups, and between the total and partial tumor resection groups. The ratio of sellar floor opening area to maximal tumor area was significantly different between the total and partial resection groups. Logistic regression analysis revealed that the ratio of sellar floor opening area to the largest tumor area, tumor texture, tumor invasion and age were independent prognostic factors. The vertical distance between the top point of sellar floor opening and planum sphenoidale significantly differed between the patients with and without leakage of cerebrospinal fluid. CONCLUSION: These results together indicated that relatively insufficient sellar floor opening is a cause of leading to residual tumor, and the higher position of the opening and closer to the planum sphenoidale are likely to induce the occurrence of leakage of cerebrospinal fluid.

Repeated transsphenoidal surgery for resection of pituitary adenoma.

To investigate the surgical strategy of repeated microscopic transsphenoidal surgery (TSS) for treatment of pituitary adenoma, surgical techniques and treatment outcomes for 29 patients with pituitary adenoma were reviewed and analyzed. There were 17 patients who underwent TSS 18 times and 12 patients who underwent TSS 13 times. The interval between each TSS ranged from 3 months to 18 years, with a median time of 4 years. The tumor height was 15 to 45 mm on the last surgery. Among the 29 patients, 16 patients underwent total tumor resection, 11 patients underwent subtotal resection, and 2 patients underwent partial resection. Cerebrospinal fluid leak occurred in 10 patients. Among 24 patients who were followed up effectively, 1 patient developed abducens paralysis after surgery, 1 patient had chronic diabetes insipidus, and 1 patient received steroid-dependent alternative treatment. The repeated TSS may present satisfied outcomes in experienced hands. The upper edge of the posterior choanae should be identified to ensure the right orientation. The openings of the anterior wall of the sphenoid sinus and the sellar floor should be appropriately expanded to improve tumor exposure. The artificial materials should be identified and removed carefully. Intraoperative cerebrospinal fluid leakage should be managed well.

The long non-coding RNA PANDAR regulates cell proliferation and epithelial-to-mesenchymal transition in glioma.

Glioma is one of the most aggressive intracranial tumors in the central nervous system. The long non-coding RNA P21 associated ncRNA DNA damage activated (PANDAR) has been reported to be an oncogene or tumor suppressor in several cancers. However, the prognostic value and biological function of PANDAR in glioma have not been described. Here, we report that expression of PANDAR is significantly up-regulated in glioma tissues and cell lines. PANDAR expression was correlated with tumor size (p=0.044) and World Health Organization (WHO) grades (p=0.005), as shown by chi-squared test. Moreover, significant upregulation of PANDAR was found to correlate with poor prognosis in glioma, as shown using Kaplan-Meier method and Cox multivariate survival analysis. Furthermore, PANDAR knockdown suppressed cell proliferation, G1/S transition, migration and invasion, and promoted apoptosis in glioma cell lines (U251 and U87). PANDAR knockdown decreased expression of CDK4, Bcl-2, N-cadherin and Vimentin, but increased E-cadherin expression in glioma cells. In conclusion, our data suggest PANDAR as a potential prognostic biomarker and therapeutic candidate for glioma.

WDR62 variants contribute to congenital heart disease by inhibiting cardiomyocyte proliferation.

BACKGROUND: Congenital heart disease (CHD) is the most common birth defect and has high heritability. Although some susceptibility genes have been identified, the genetic basis underlying the majority of CHD cases is still undefined. METHODS: A total of 1320 unrelated CHD patients were enrolled in our study. Exome-wide association analysis between 37 tetralogy of Fallot (TOF) patients and 208 Han Chinese controls from the 1000 Genomes Project was performed to identify the novel candidate gene WD repeat-containing protein 62 (WDR62). WDR62 variants were searched in another expanded set of 200 TOF patients by Sanger sequencing. Rescue experiments in zebrafish were conducted to observe the effects of WDR62 variants. The roles of WDR62 in heart development were examined in mouse models with Wdr62 deficiency. WDR62 variants were investigated in an additional 1083 CHD patients with similar heart phenotypes to knockout mice by multiplex PCR-targeting sequencing. The cellular phenotypes of WDR62 deficiency and variants were tested in cardiomyocytes, and the molecular mechanisms were preliminarily explored by RNA-seq and co-immunoprecipitation. RESULTS: Seven WDR62 coding variants were identified in the 237 TOF patients and all were indicated to be loss of function variants. A total of 25 coding and 22 non-coding WDR62 variants were identified in 80 (6%) of the 1320 CHD cases sequenced, with a higher proportion of WDR62 variation (8%) found in the ventricular septal defect (VSD) cohort. WDR62 deficiency resulted in a series of heart defects affecting the outflow tract and right ventricle in mouse models, including VSD as the major abnormality. Cell cycle arrest and an increased number of cells with multipolar spindles that inhibited proliferation were observed in cardiomyocytes with variants or knockdown of WDR62. WDR62 deficiency weakened the association between WDR62 and the cell cycle-regulated kinase AURKA on spindle poles, reduced the phosphorylation of AURKA, and decreased expression of target genes related to cell cycle and spindle assembly shared by WDR62 and AURKA. CONCLUSIONS: WDR62 was identified as a novel susceptibility gene for CHD with high variant frequency. WDR62 was shown to participate in the cardiac development by affecting spindle assembly and cell cycle pathway in cardiomyocytes.

Magnetic resonance imaging characteristics of residual pituitary tissues following transsphenoidal resection of pituitary macroadenomas.

OBJECTIVE: The present study is to investigate the pre- and post-operative magnetic resonance imaging of pituitary tissues following transsphenoidal resection of pituitary macroadenomas, as well as its clinical significance. MATERIALS AND METHODS: The medical records of 108 consecutive pituitary macroadenoma patients admitted at Fuzhou 900th Hospital between September 2012 and September 2014 were retrospectively reviewed. Siemens 3. 0T magnetic resonance scanner was used to perform pre- and postoperative MRI scanning, including plain scan and contrast-enhanced scan of SE sequential T1WI and T2WI in sagittal, coronal and axial views. PACS medical imaging system was used to measure the diameter of pituitary adenoma, as well as the volumes of the adenoma and pituitary tissue. Hematoxylin-eosin staining and immunohistochemical staining were also performed. RESULTS: Higher height of pituitary adenoma results in lower rate of posterior pituitary bright spot (PPBS) on MR T1-weighted imaging. Preoperative MR signal intensity of PPBS was negatively related to diabetes insipidus (DI). Normal pituitary tissues were likely to be above the pituitary adenomas in growth hormone-secreting adenoma patients, while mostly located aside in gonadotropin-secreting adenoma patients. Morphological restitution of postoperative pituitary tissues was better in lateral displacement than that in superior or superolateral patterns on pre-operative MR images. Positive rate of PPBS on preoperative MRI is negatively related to adenoma height, and the signal intensity of PPBS is inversely related to postoperative DI. CONCLUSIONS: The relative locations of pituitary tissues and adenoma tissues may be associated with the adenoma type and may affect the postoperative remodeling of residual pituitary tissues.

The Emerging Roles of the RNA Binding Protein QKI in Cardiovascular Development and Function.

RNA binding proteins (RBPs) have a broad biological and physiological function and are critical in regulating pre-mRNA posttranscriptional processing, intracellular migration, and mRNA stability. QKI, also known as Quaking, is a member of the signal transduction and activation of RNA (STAR) family, which also belongs to the heterogeneous nuclear ribonucleoprotein K- (hnRNP K-) homology domain protein family. There are three major alternatively spliced isoforms, QKI-5, QKI-6, and QKI-7, differing in carboxy-terminal domains. They share a common RNA binding property, but each isoform can regulate pre-mRNA splicing, transportation or stability differently in a unique cell type-specific manner. Previously, QKI has been known for its important role in contributing to neurological disorders. A series of recent work has further demonstrated that QKI has important roles in much broader biological systems, such as cardiovascular development, monocyte to macrophage differentiation, bone metabolism, and cancer progression. In this mini-review, we will focus on discussing the emerging roles of QKI in regulating cardiac and vascular development and function and its potential link to cardiovascular pathophysiology.

The Role of Tbx20 in Cardiovascular Development and Function.

Tbx20 is a member of the Tbx1 subfamily of T-box-containing genes and is known to play a variety of fundamental roles in cardiovascular development and homeostasis as well as cardiac remodeling in response to pathophysiological stresses. Mutations in TBX20 are widely associated with the complex spectrum of congenital heart defects (CHDs) in humans, which includes defects in chamber septation, chamber growth, and valvulogenesis. In addition, genetic variants of TBX20 have been found to be associated with dilated cardiomyopathy and heart arrhythmia. This broad spectrum of cardiac morphogenetic and functional defects is likely due to its broad expression pattern in multiple cardiogenic cell lineages and its critical regulation of transcriptional networks during cardiac development. In this review, we summarize recent findings in our general understanding of the role of Tbx20 in regulating several important aspects of cardiac development and homeostasis and heart function.

Early onset developmental delay and epilepsy in pediatric patients with WDR45 variants.

BACKGROUND: Developmental delay (DD) is a neurological disorder that presents with defects in gross motor, fine motor, language and cognition functions. WD repeat domain 45 (WDR45) is one of the disease-causing genes of DD. Previously, WDR45 de novo mutations were reported in certain adult and pediatric patients due to iron accumulation. CLINICAL REPORT: We report five pediatric female patients with DD and epilepsy. Their ages were below 3 years at the first consultation, and precise diagnoses were difficult based on the available clinical information and phenotype. METHODS: Children with DD and/or epilepsy presenting to the molecular diagnostic center of Children's Hospital of Fudan University between May 2016 and May 2017 were enrolled. The patients and their parents were subjected to whole-exome sequencing (WES), and we characterized the phenotypes of the patients carrying WDR45 variants. Furthermore, we overexpressed the candidate variants in HeLa cells and evaluated their effect on autophagy through Western blot and immunofluorescence staining with confocal microscopy. RESULTS: Five WDR45 de novo mutations, namely, c.19C > T (p.Arg7*), c.401G > C (p.Arg134Pro), c.503G > A (p.Gly168Glu), c.700C > T (p.Arg234*), and c.912delT (p.Ala305Leufs*25), were detected in 623 enrolled pediatric patients (274 females; 487 patients younger than 6 years). All five patients with WDR45 variants presented with DD and epilepsy. Compared with the control HeLa cells, the cells with the p. Arg134Pro and p. Gly168Glu missense mutations showed accumulation of LC3-containing autophagic structures and an abnormally enlarged cell volume, and Western blotting revealed a significant increase in LC3II/GAPDH. CONCLUSION: The identification of WDR45 mutations provides further evidence that WES plays an important role in the diagnosis of neurological disorders with common phenotypes and that WDR45 mutations are associated with neurological disorders and are not very rare in Chinese female pediatric patients with DD and/or epilepsy. The diagnosis of patients with WDR45 mutations would enable more precise genetic counseling for the parents of these children.

Role of prolactin/adenoma maximum diameter and prolactin/adenoma volume in the differential diagnosis of prolactinomas and other types of pituitary adenomas.

The present study aimed to investigate the function of the prolactin/adenoma maximum diameter (PRL/MD) and the prolactin/adenoma volume (PRL/V) in the differential diagnosis of prolactinomas and other types of pituitary adenomas. A total of 118 patients with pituitary adenoma, hyperprolactinemia and a plasma PRL <250 µg/l were enrolled. Clinical data from these patients were retrospectively analyzed. A receiver operating characteristic curve was plotted. The function of PRL, PRL/MD and PRL/V in the differential diagnosis of prolactinomas and other types of pituitary adenomas was compared. The results revealed that a PRL of 55.65 µg/l was the most accurate [sensitivity (SE), 0.800; specificity (SP), 0.716; positive predictive value (PPV), 0.857; negative predictive value (NPV), 0.933; and Youden index (YI), 0.516]. The PRL/MD with the highest diagnostic value was 4.03 µg/(l × mm) (SE, 0.800; SP, 0.898; PPV, 0.727; NVP, 0.929; and YI, 0.698). The PRL/V with the highest diagnostic value was 54.00 µg/(l × cm3) (SE, 0.900; SP, 0.966; PPV, 0.900; NVP, 0.966; and YI, 0.866). The PRL/MD tended to be of higher diagnostic accuracy than PRL, but this difference was not statistically significant (P=0.097). The differentiation ability of PRL/V was significantly stronger than that of PRL (P=0.028). Thus, serum PRL, PRL/MD and PRL/V levels may be able to differentiate prolactinomas from other types of hyperprolactinemia-causing pituitary adenomas prior to treatment. PRL/V may be better than the PRL level in achieving a differential diagnosis, and the optimal PRL/V ratio for differentiating prolactinomas from other types of hyperprolactinemia-causing pituitary adenomas was 54.00 µg/(l × cm3).

Imaging Evaluation of the Location and Fenestration of Sellar Floor During Endonasal Transsphenoidal Surgery in Patients with Pituitary Adenomas.

OBJECTIVE: To evaluate clinical value of three-dimensional (3D) computed tomography (CT) reconstruction of the sphenoid sinus separation in localizing sellar floor during endonasal transsphenoidal surgery and determine size and location of sellar floor fenestration. METHODS: There were 51 patients eligible for study inclusion. Preoperative CT scan of the paranasal sinus and CT scan and magnetic resonance imaging of the pituitary gland were obtained. Sphenoid sinus separation was reconstructed using Mimics 15.0 software, and quantity, shape, and orientation were observed and compared with intraoperative data to guide the localization of sellar floor. Anatomic variation of the sphenoid sinus and adjacent structures, tumor and sella turcica morphology, minimal distance between the cavernous segment of the internal carotid artery bilaterally, and shortest distance from the midline were measured. RESULTS: Based on the shape of the sphenoid sinus separation, sellar floor was accurately localized in all cases. Intraoperative sphenoid sinus separation was consistent with preoperative three-dimensional CT reconstruction images. The sellar floor was extremely small in 2 patients, and insufficient fenestration of sellar floor negatively affected tumor resection. Preoperative three-dimensional CT reconstruction is helpful for accurate and rapid localization of sellar floor. CONCLUSIONS: Anatomic variation of sphenoid sinus and adjacent structures, characteristics of tumor and sella, minimum distance between bilateral cavernous segment of the internal carotid artery, and shortest distance from midline are helpful for establishment of individualized sellar floor fenestration.