Xianmin Xiao

Effects of propranolol on the proliferation and apoptosis of hemangioma-derived endothelial cells.

BACKGROUND/PURPOSE Propranolol, a non-selective beta-blocker, has recently been introduced as a novel treatment modality for proliferating hemangiomas. However, the mechanism of action of this therapy is unknown. In this study, we investigated propranolol in the etiology of hemangiomas that are associated with the proliferation and apoptosis of hemangioma-derived endothelial cells (HemECs). METHODS HemECs were isolated from freshly resected hemangioma specimens. We studied propranolol-treated HemECs in vitro. We measured the effect of propranolol on HemEC viability using the Cell Counting Kit-8 (CCK-8) assay and proliferation and apoptosis using a BrdU labeling assay, annexin-V-fluorescein isothiocyanate/propidium iodide flow cytometry, and Hoechst 33342 fluorescent staining. We explored the potential mechanisms of propranolol-induced HemEC dysfunction using western blot analysis, a caspase assay kit, and real-time quantitative PCR. RESULTS We observed that propranolol had inhibitory effects on the viability and proliferation of HemECs. HemEC apoptosis significantly increased with 100 μM propranolol treatment. Vascular endothelial growth factor (VEGF) expression was down-regulated by propranolol in a dose-dependent manner. We also demonstrated activation of the caspase cascade, including caspase-9 and caspase-3 of the intrinsic pathway, and an increased p53 gene expression and Bax/Bcl-xL ratio in HemECs treated with 100 μM propranolol. CONCLUSIONS We obtained novel data that suggests propranolol could inhibit HemEC proliferation and induce apoptosis. The effects were likely mediated through the suppression of VEGF expression, activation of caspase-9 and caspase-3, up-regulation of the pro-apoptotic genes p53 and Bax and down-regulation of the anti-apoptotic gene Bcl-xL.

The role of β-adrenergic receptor signaling in the proliferation of hemangioma-derived endothelial cells

BackgroundInfantile hemangioma (IH) is a benign vascular neoplasm that arises from the abnormal proliferation of endothelial cells and enhanced angiogenesis. Recently, propranolol has been found to be effective in the management of IH, suggesting that β-adrenergic receptors (β-ARs) may play an important role in the pathogenesis of IH.ResultsIn the present study, we investigated the β-adrenergic signaling that is associated with hemangioma-derived endothelial cell (HemEC) proliferation. The results showed that both β1- and β2-ARs were expressed in HemECs. Stimulation of the β-ARs by isoprenaline induced cell proliferation and elevation of second messenger cAMP levels. The proliferation-promoting action of isoprenaline was abolished by a β1-selective antagonist and was more effectively abolished by a β2-selective antagonist; the mechanism for the action of the antagonists was a G0/G1 phase cell cycle arrest which was associated with decreased cyclin D1, CDK-4, CDK-6 and phospho-Rb expression. Pre-treatment of the cells with VEGFR-2 or ERK inhibitors also prevented the isoprenaline-mediated proliferation of cells. In agreement with the involvement of β-ARs and VEGFR-2 in the HemEC response, β-AR antagonists and the VEGFR-2 inhibitor significantly attenuated isoprenaline-induced ERK phosphorylation. Moreover, treating the cells with isoprenaline markedly increased VEGF-A expression and VEGFR-2 activity in a β2-AR-dependent manner.ConclusionsWe have demonstrated that the activation of the β-ARs in the ERK pathway may be important mechanisms in promoting HemEC growth. Furthermore, stimulation of the β-AR may transactivate VEGFR-2 signaling and further increase HemEC proliferation.

Growth-promoting effect of bisphenol A on neuroblastoma in vitro and in vivo

PURPOSE To investigate the effect and mechanism of bisphenol A (BPA), one of the main environmental endocrine disruptors, on the proliferation of human neuroblastoma cells. METHODS In vitro, cultured SK-N-SH cells were treated with 17beta-estradiol (E(2); 1 ng/mL), BPA (2 microg/mL) with or without estrogen receptor antagonist ICI182,780 (10(-6) mol/L). Viable cell number, DNA proliferation index, and expression of cyclin-dependent kinase 4 and cyclin D1 were assessed by absorbance reading, flow cytometry, and western blotting, respectively. In vivo, ovariectomized nude mice bearing SK-N-SH tumors were administered by gavage with E(2) (500 microg/kg per day, n = 11), BPA (200 mg/kg per day, n = 10), or vehicle (n = 9) for 18 days. Mice body weight, tumor volume and weight were examined every 3 days. Tumor microvessel density, proliferating cell nuclear antigen and vascular endothelial growth factor expression were evaluated by immunohistochemical staining or western blotting. RESULTS In vitro, the BPA group had 20% higher number of viable cells, 70% higher proliferation index (both P < .01), and higher expression of cyclin-dependent kinase 4 and cyclin D1 than the nontreated group. In vivo, the BPA group had over 50% higher gross tumor volume, tumor weight, microvessel density, proliferating cell nuclear antigen (P < .05 or .01), and higher vascular endothelial growth factor protein expression than the mock control group. Both in vitro and in vivo BPA effects were comparable with those by E(2). ICI182,780 effectively abolished the promoting effect for both. CONCLUSIONS Bisphenol A can promote the growth of neuroblastoma to a level similar to that of E(2). Estrogen receptor-dependent pathway and angiogenesis may contribute to the underlying mechanisms.

Comment on: Steroid-resistant Kaposiform Hemangioendothelioma: A retrospective study of 37 patients treated with vincristine and long-term follow-up

To the Editor: We read with great interest the report by Wang et al. (Pediatr Blood Cancer 2015; 62:577–580)[1] providing a retrospective review of 37 cases of Kaposiform Hemangioendothelioma (KHE) treated with vincristine, and the accompanying highlight by Francine Blei, pages 551–552).[2] Evidence-based management of vascular anomalies has been impeded by a dearth of prospective clinical trials. Reasons include a lack of a phenotype/genotype classification system, limited numbers of specialists with clinical trial acumen, a historical slant toward surgical and interventional treatments, a paucity of funding options, and rarity of the individual diagnoses. These deficits have pushed patients and families to rally together to demand improved clinical and basic science research and expertise. Presently, the field of vascular anomalies is flourishing with a recently expanded classification system[3] as well as new prospective clinical trials. We believe that collaborative clinical trials among multidisciplinary disease specialists are mandatory for vascular anomalies to emulate the advances made in pediatric oncology by the Children’s oncology group. KHE is a vascular tumor treated by Pediatric Hematologists/ Oncologists. Vincristine treatment is the current consensus-derived treatment for this tumor.[4] After compassionate use of sirolimus for one patient, Cincinnati Children’s Hospital Medical Center (CCHMC) prospectively studied sirolimus as treatment in a total of 60 patients with complicated vascular anomalies.[5] This phase 2 study (NCT# 00975819) was completed in 2014, presented at ISSVA and ASPHO, and recently submitted for publication. The clinical trial proved sirolimus to be safe and efficacious for a majority of vascular anomalies, with 13 of 13 participants with KHE with KMP having partial responses. Preliminary results with sirolimus appear favorable but many questions remain unanswered, including effectiveness compared to standard vincristine-based therapy, time to response, durability of response, and long-term effects. The present retrospective study[1] and the expert opinion consensus paper[4] recommend vincristine and steroids as the first line treatment for KHEwith KMP. Dr. Blei’s thoughtful highlight article[2] provides historical perspective on KHE therapy, but concludes with the suggestion of sirolimus as a first line agent. We suggest caution before considering sirolimus as first line therapy based on small numbers of patients. We strongly recommend that these questions be answered in a randomized controlled trial. This study actually recently opened at CCHMC (NCT # 02110069—“A Randomized Phase 2 Study of Vincristine vs. Sirolimus to treat High Risk Kaposiform Hemangioendothelioma [KHE]”). This FDA-funded and Pfizer-supported trial will open at seven additional sites throughout the United States in coming months, establishing an efficient referral network. This study will establish the most efficacious treatment for KHE with KMP, incorporatingmulti-faceted outcomemonitoring, long-term followup, and the use of adaptive trial design with early stopping rules. Collaborative studies are needed for many of the disease entities within the field of vascular anomalies. Pediatric hematologist/ oncologists are taking leadership roles in the treatment of these diseases. ThroughASPHO, theVascular Anomalies Special Interest Group has drawn tremendous interest and is one example of a collaborative group forming within our specialty to utilize our clinical research expertise to improve outcomes for patients with vascular anomalies.

Endoscopic-assisted surgery for pyriform sinus fistula in children: Experience of 165 cases from a single institution

BACKGROUND Congenital pyriform sinus fistula (PSF) is relatively rare, but often presents diagnostic and therapeutic challenges. Herein, we report our experience of endoscopic-assisted surgery of PSF in children. METHODS Since 1999, 165 children (100 males, 65 females) with PSF had been enrolled. Their clinical manifestations were recurrent lateral neck infection, cervical mass and respiratory distress. Preoperative investigations included barium swallow, ultrasound, computed tomography, and thyroid scan. After resolution of the infection, the fistulas were identified by the endoscopic-assisted technique at operation. The fistula tract was completely excised just at the apex of the pyriform sinus. More recently, the anatomic point where the fistula tract penetrated into the pharynx was specified and recorded. RESULTS Of the 165 cases, the male to female ratio was 1.54:1, the median age of onset was 3.2 years (range, 1 day to 13.8 years), and median age at operation was 5.0 years (range, 17 days to 15.0 years). One hundred fifty-six (94.6%) fistulas located on the left side, 7 right, and 2 bilateral. Twelve neonates and young infants (younger than 3 months) presented with a large cervical cyst. The fistulas were completely excised in all but 2 (98.8%) with intraoperative gastroscopy successfully conducted in 160 cases (97.0%). In 77 cases the points where fistulas penetrated into the pharynx were specified intraoperatively, which were classified into 3 types according to their anatomic relationship with the inferior cornu of the thyroid cartilage (ICTC): type I (anterior to ICTC), 22 cases (28.5%); type II (inferior to ICTC), 18 cases (23.4%); and type III (posterior to ICTC), 37 cases (48.1%). Postoperatively, 160 cases recovered well without complications. PSF recurred in 5 cases, 2 of whom were cured by fistula re-excision and 3 remained asymptomatic. Esophageal perforation was found and repaired uneventfully in 1 neonate and 1 young infant. Transient postoperative hoarseness happened in 1 neonate. All the latter 3 cases had cervical cysts. CONCLUSIONS To our knowledge, this series is the largest report of PSF in children. Our results suggested that PSF is more common in males. With the help of endoscopy and a better understanding of the anatomic relationship between ICTC and the points where the fistulas penetrated into the pharynx, PSF excision can be done successfully with minimized complications. For neonates and young infants with a cervical cyst, however, the management of PSF continues to be a challenge.

Steroid-resistant kaposiform hemangioendothelioma: A retrospective study of 37 patients treated with vincristine and long-term follow-up: Vincristine for Steroid-Resistant KHE

Kaposiform hemangioendothelioma (KHE) with Kasabach–Merritt phenomenon (KMP) still remains a particular and life‐threatening disease. The purpose of this study was to evaluate the efficacy of vincristine (VCR) and the possibility of replacement with steroids in the treatment of steroid‐resistant KHE with KMP.

Upregulated autocrine vascular endothelial growth factor (VEGF)/VEGF receptor-2 loop prevents apoptosis in haemangioma-derived endothelial cells

The autocrine vascular endothelial growth factor (VEGF)/VEGF receptor (VEGFR)‐2 loop is required to maintain the transformed phenotype of many tumours, in part, by preventing apoptotic cell death in response to many different stimuli. However, it is unclear whether constitutive VEGF/VEGFR‐2 activation in haemangioma‐derived endothelial cells (HaemECs) can lead to a general suppression of apoptosis.

Diagnosis and management of pyriform sinus fistula: Experience in 48 cases ☆

PURPOSE Pyriform sinus fistula (PSF) is often overlooked, and presents diagnostic and management challenge. The aim of this study was to highlight the value of intraoperative endoscopy-assisted intubation or methylene blue injection through the internal opening as a guide in searching for the fistula. METHODS The charts of 48 patients diagnosed with PSF during January 1990 until January 2013 were retrospectively reviewed. The records were analyzed for sex, side of lesion, age at onset/diagnosis, initial presentations, diagnostic methods, microbiologic cultures, pathologic findings, treatments and outcomes. RESULTS There were 22 males and 26 females, with a median age at onset and diagnosis of 2 years (range, 8 months to 9 years) and 4 years (range, 12 months to 13 years), respectively. The lesions were predominantly left sided (93.7%). The most common presentation was neck abscess (62.5%). Other presentations were acute suppurative thyroiditis/thyroid abscess (7), neck mass with or without dyspnea (9), and thyroid nodule (2). Barium esophagography showed the sinus tract in 100% cases. The positive predictive value of other modalities was oral-contrast CT 88.9%, intravenous contrast-enhanced CT 53.8%, noncontrast CT 33.3%, and sonography 7.9%. Thyroid function were reported normal in most tested cases (14/15, 93.3%). The fistula tract was lined with pseudostratified squamous epithelium or ciliated columnar epithelium, often associated with inflammatory changes. Bacteria cultured from the discharge were found to be oral flora. Partial thyroidectomy was operated on 11 cases. Two patients (2/8, 25%) who underwent open surgery without endoscopic assistance exhibited recurrence, while no recurrence was noted in children with the help of intraoperative endoscopy. Postoperative results were good in majority (93.7%). CONCLUSION PSF should be considered in any children with repeated neck or thyroid infection/mass. The combination of barium esophagography, CT scan and ultrasound is useful to establish the diagnosis. Intraoperative endoscopy-assisted intubation or methylene blue injection through the internal opening as a guide can facilitate identification of the tract during dissection.

Measuring health-related quality of life in children with cancer living in mainland China: feasibility, reliability and validity of the Chinese mandarin version of PedsQL 4.0 Generic Core Scales and 3.0 Cancer Module

BackgroundThe Pediatric Quality of Life Inventory (PedsQL) is widely used instrument to measure pediatric health-related quality of life (HRQOL) for children aged 2 to 18 years. The purpose of the current study was to investigate the feasibility, reliability and validity of the Chinese mandarin version of the PedsQL 4.0 Generic Core Scales and 3.0 Cancer Module in a group of Chinese children with cancer.MethodsThe PedsQL 4.0 Genetic Core Scales and the PedsQL 3.0 Cancer Module were administered to children with cancer (aged 5-18 years) and parents of such children (aged 2-18 years). For comparison, a survey on a demographically group-matched sample of the general population with children (aged 5-18) and parents of children (aged 2-18 years) was conducted with the PedsQL 4.0 Genetic Core Scales.ResultThe minimal mean percentage of missing item responses (except the School Functioning scale) supported the feasibility of the PedsQL 4.0 Generic Core Scales and 3.0 Cancer Module for Chinese children with cancer. Most of the scales showed satisfactory reliability with Cronbachs α of exceeding 0.70, and all scales demonstrated sufficient test-retest reliability. Assessing the clinical validity of the questionnaires, statistically significant difference was found between healthy children and children with cancer, and between children on-treatment versus off-treatment ≥12 months. Positive significant correlations were observed between the scores of the PedsQL 4.0 Generic Core Scale and the PedsQL 3.0 Cancer Module. Exploratory factor analysis demonstrated sufficient factorial validity. Moderate to good agreement was found between child self- and parent proxy-reports.ConclusionThe findings support the feasibility, reliability and validity of the Chinese Mandarin version of PedsQL 4.0 Generic Core Scales and 3.0 Cancer Module in children with cancer living in mainland China.

A follow-up study on postoperative function after a transanal Soave 1-stage endorectal pull-through procedure for Hirschsprung's disease

OBJECTIVE The aim of the study is to appraise bowel movements in children with Hirschsprungs disease (HD) after a transanal Soave 1-stage endorectal pull-through (TAS) procedure. METHODS From October 2000 to October 2004, 44 patients with HD underwent a TAS procedure, 35 had a Soave operation via a laparotomy, 29 underwent a Soave procedure via laparoscopy, and 39 had an Ikeda-Soper procedure; the patients were followed up from 1 to 5 years after surgery. Evaluation of the perioperative therapeutic effect and postoperative bowel movements between the 4 groups were analyzed by Kellys score and anorectal manometry. RESULTS The TAS procedure required less operative time and costs than the Soave procedure via laparotomy or laparoscopy (P < .05) and less than that of Ikeda-Soper procedure for the duration of the preoperative preparation, the duration of the surgical procedure, intraoperative blood transfusions, days of postoperative fasting, antibiotic use, and days and cost of hospitalization (P < .01). There were no differences in short-term and long-term complications between the same Soave procedures; the Soave procedure exceeded the Ikeda-Soper operation in the incidence of enterocolitis 3 months postoperatively (P < .05), but there was no difference between the TAS procedure and the Ikeda-Soper procedure. There was no difference in bowel movements 12 months postoperatively and the rectal anal inhibitory reflex, high-pressure zone length, resting anal canal pressure, and the sensation threshold 1 year postoperatively between the TAS procedure and the Ikeda-Soper procedure, but the active contractile pressure was lower after the TAS procedure than the Ikeda-Soper procedure. CONCLUSIONS The TAS procedure is more suitable than the Soave operation via laparotomy or laparoscopy and Ikeda-Soper procedure and is feasible in infants with short segment type and common type HD.