Classification and management strategies for paediatric chronic nonbacterial osteomyelitis and chronic recurrent multifocal osteomyelitis.

INTRODUCTION: Chronic non-bacterial osteomyelitis (CNO) is an autoinflammatory bone disease that most commonly affects children and adolescents causing significant pain and damage to bones. The absence of diagnostic criteria and biomarkers, an incomplete understanding of the molecular pathophysiology, and lack of evidence from randomized and controlled trials make the diagnosis and care challenging. AREAS COVERED: This review provides an overview of the clinical and epidemiological features of CNO and displays diagnostic challenges and how they can be addressed following strategies used internationally and by the authors. It summarizes the molecular pathophysiology, including pathological activation of the NLRP3 inflammasome and IL-1 secretion, and how these observations can inform future treatment strategies. Finally, it provides a summary of ongoing initiatives aiming at classification criteria (ACR/EULAR) and outcome measures (OMERACT) that will enable the generation of evidence through clinical trials. EXPERT OPINION: Scientific efforts have linked molecular mechanisms to cytokine dysregulation in CNO, thereby delivering arguments for cytokine blocking strategies. Recent and ongoing collaborative international efforts are providing the basis to move toward clinical trials and target directed treatments for CNO that find approval by regulatory agencies.

The transition zone in Hirschsprung's bowel contains abnormal hybrid ganglia with characteristics of extrinsic nerves.

The aganglionic bowel in short-segment Hirschsprung's disease is characterized both by the absence of enteric ganglia and the presence of extrinsic thickened nerve bundles (TNBs). The relationship between the TNBs and the loss of enteric ganglia is unknown. Previous studies have described decreasing numbers of ganglia with increasing density of TNBs within the transition zone (TZ) between ganglionic and aganglionic gut, and there is some evidence of spatial contact between them in this region. To determine the cellular interactions involved, we have analysed the expression of perineurial markers of TNBs and enteric ganglionic markers for both neural cells and their ensheathing telocytes across four cranio-caudal segments consisting of most proximal ganglionic to most distal aganglionic from pull-through resected colon. We show that in the TZ, enteric ganglia are abnormal, being surrounded by perineurium cells characteristic of TNBs. Furthermore, short processes of ganglionic neurons extend caudally towards the aganglionic region, where telocytes in the TNB are located between the perineurium and nerve fibres into which they project telopodes. Thus, enteric ganglia within the TZ have abnormal structural characteristics, the cellular relationships of which are shared by the TNBs. These findings will help towards elucidation of the cellular mechanisms involved in the aetiology of Hirschsprung's disease.

Thyroid cancer in children: A multicenter international study highlighting clinical features and surgical outcomes of primary and secondary tumors.

Background: Thyroid gland malignancies are rare in pediatric patients (0.7% of tumors); only 1.8% are observed in patients aged <20 years, with a higher prevalence recorded in women and adolescents. Risk factors include genetic syndromes, MEN disorders, autoimmune diseases, and exposure to ionizing radiation. Radiotherapy is also associated with an increased risk of secondary thyroid cancer. This study describes the clinical features and surgical outcomes of primary and secondary thyroid tumors in pediatric patients. Methods: Institutional data were collected from eight international surgical oncology centers for pediatric patients with thyroid cancer between 2000 and 2020. Statistical analyses were performed using the GraphPad Prism software. Results: Among 255 total cases of thyroid cancer, only 13 (5.1%) were secondary tumors. Primary thyroid malignancies were more likely to be multifocal in origin (odds ratio [OR] 1.993, 95% confidence interval [CI].7466-5.132, p = 0.2323), have bilateral glandular location (OR 2.847, 95% CI.6835-12.68, p = 0.2648), and be metastatic at first diagnosis (OR 1.259, 95% CI.3267-5.696, p > 0.999). Secondary tumors showed a higher incidence of disease relapse (OR 1.556, 95% CI.4579-5.57, p = 0.4525) and surgical complications (OR 2.042, 95% CI 0.7917-5.221, p = 0.1614), including hypoparathyroidism and recurrent laryngeal nerve injury. The overall survival (OS) was 99% at 1 year and 97% after 10 years. No EFS differences were evident between the primary and secondary tumors (chi-square 0.7307, p = 0.39026). Conclusions: This multicenter study demonstrated excellent survival in pediatric thyroid malignancies. Secondary tumors exhibited greater disease relapse (15.8 vs. 10.5%) and a higher incidence of surgical complications (36.8 vs. 22.2%).

SARS-COV2 placentitis and pregnancy outcome: A multicentre experience during the Alpha and early Delta waves of coronavirus pandemic in England.

Background: Pregnant women with SARS-CoV-2 infection experience higher rates of stillbirth and preterm birth. A unique pattern of chronic histiocytic intervillositis (CHI) and/or massive perivillous fibrin deposition (MPFD) has emerged, coined as SARS-CoV-2 placentitis. Methods: The aim of this study was to describe a cohort of placentas diagnosed with SARS-CoV-2 placentitis during October 2020-March 2021. Cases with a histological diagnosis of SARS-CoV-2 placentitis and confirmatory immunohistochemistry were reported. Maternal demographic data, pregnancy outcomes and placental findings were collected. Findings: 59 mothers delivered 61 infants with SARS-CoV-2 placentitis. The gestational age ranged from 19 to 41 weeks with most cases (78.6%) being third trimester. 30 infants (49.1%) were stillborn or late miscarriages. Obese mothers had higher rates of pregnancy loss when compared with those with a BMI <30 [67% (10/15) versus 41% (14/34)]. 47/59 (79.7%) mothers had a positive SARS-CoV-2 PCR test either at the time of labour or in the months before, of which 12 (25.5%) were reported to be asymptomatic. Ten reported only CHI, two cases showed MPFD only and in 48 placentas both CHI and MPFD was described. Interpretation: SARS-CoV2 placentitis is a distinct entity associated with increased risk of pregnancy loss, particularly in the third trimester. Women can be completely asymptomatic and still experience severe placentitis. Unlike 'classical' MPFD, placentas with SARS-CoV-2 are generally normal in size with adequate fetoplacental weight ratios. Further work should establish the significance of the timing of maternal SARS-CoV-2 infection and placentitis, the significance of SARS-CoV2 variants, and rates of vertical transmission associated with this pattern of placental inflammation. Funding: There was not funding associated with this study.

A Quantitative Assessment of Mucosal Eosinophils in the Gastrointestinal Tract of Children Without Detectable Organic Disease.

BACKGROUND: Accurate measurements of mucosal eosinophil concentrations in gastrointestinal tracts of healthy children are necessary to differentiate health and disease states in general, and better define eosinophilic gastrointestinal diseases. STUDY: We retrospectively reviewed gastrointestinal biopsies from children with macroscopically normal endoscopies, who, after a minimal follow-up of one year, were not diagnosed with any organic disease. Peak eosinophil concentrations and distributions were assessed from each segment of the gastrointestinal tract. RESULTS: Three centers (Italy, United Kingdom, and Israel) contributed 202 patients (median age 13 years IQR 9.5-15.5, range 1-18 years). Median (IQR, range) eosinophil concentrations (eos/mm2) were: esophagus 0 (0-0, 0-84), stomach 0 (0-4, 0-84), duodenal bulb 20 (13-30, 7-67), second part of duodenum 20 (13-29, 0-105), terminal ileum 29 (14-51, 0-247), cecum 53 (37-89, 10-232), ascending colon 55 (25-84, 0-236), transverse colon 38 (21-67, 4-181), descending colon 29 (17-59, 0-114), sigmoid colon 25 (13-40, 0-215) and rectum 13 (4-28, 0-152). Significant geographical variance was present, however, no differences in eosinophil concentrations were identified between children with resolving symptoms vs. those with functional diagnoses, nor across age groups. CONCLUSIONS: Standardized eosinophil concentrations from the gastrointestinal tracts of children without organic disease will serve to better define both health and disease states. No differences were found between resolved symptoms vs. functional diagnoses nor between age groups in this pediatric cohort.

Isotope and hydrochemical systematics of groundwater from a multi-tiered aquifer in the central parts of Indo-Gangetic Plains, India - Implications for groundwater sustainability and security.

The Indo-Gangetic multi-aquifer system provides water supplies to the most populous regions of the Indian subcontinent, however precise knowledge on the sources and dynamics of groundwater is still missing. Environmental isotopes (2H, 18O, 13C, 3H and 14C) and hydrochemical modeling tools were used in this study in the multi-tiered aquifers underlying the Middle Gangetic Plains (MGP) to investigate the source of recharge, aquifer dynamics and inter-connectivity among aquifers. Within a depth span of 300 m, three aquifers, with contrasting recharge sources and dynamics, were delineated in this Sone-Ganga-Punpun interfluve region, with limited cross-aquifer hydraulic interconnections. The chemistry evolves from Ca-HCO3 to Na-Ca-HCO3 in the shallow semiconfined Aquifer-I with a mean transit time of 20-23 years. The dominant recharge to Aquifer-I is from the river inflows and rainwater percolation through paleochannels. The semi-confined to confined Aquifer-II holds fresh quality groundwater with mixed water facies (Mg/Ca-Na-HCO3). The modeled age of Aquifer-II groundwater is found to be 205-520 years, which is supported by presence of negligible tritium and minor variations in stable isotopes. Outcrop regions of Aquifer-II sediments in the marginal alluvial areas and deep-seated paleochannels in the southwestern part are the potential zones for Aquifer-II recharge. A deep confined Aquifer-III with fresh quality of groundwater is identified below 220 m. This aquifer is characterized by old age (~3.5 to 4.7 ka BP) and enriched δ18O (-5.7‰). These results along with the existing paleoclimate records of this region infer that Aquifer-III is recharged during an arid climate. The marginal alluvial plains are the probable recharge zones for Aquifer-III. This study helped in conceptualizing the groundwater flow paths in multi-tiered aquifers of MGP. The knowledge and understanding would extend crucial inputs for the sustainable development of deep aquifers not only in the MGP but also in other regions of Indo-Gangetic Plains.

Management and outcome of paediatric testicular tumours - A 20 year experience.

OBJECTIVE: To report a 20-year experience highlighting management and outcome(s) of paediatric testicular tumours. PATIENTS AND METHODS: All males (< 19 years) with an index diagnosis of testicular tumours during the era(s) 1998-2018 in North West England were identified. Data were collected regarding age at diagnosis, disease stage, surgical operations, tumour biology and outcome(s). RESULTS: A total of 34 male patients were identified. Median age at primary diagnosis was 94 months (range: 0-229 months). Eighteen tumours were benign and 16 malignant. Twenty cases (59%) were recorded in pre pubertal children and 14 (41%) in post pubertal males . In the pre pubertal group (0-11 years) - 15 cases of germ cell tumours (unrelated to germ cell neoplasia in situ - non-GCNIS derived) were recorded, including six yolk sac lesions, eight teratomas and one mixed teratoma/yolk sac tumour (pre-pubertal type). Four males with sex cord-stromal tumours included one juvenile granulosa cell tumour, two Sertoli cell tumours and one Leydig cell tumour. One miscellaneous type tumour notably a papillary cyst adenoma was also identified. In the post pubertal male cohort (>12 years) (n = 14) - four non-GCNIS derived tumours were identified (3 epidermoid cysts and one teratoma), eight cases of germ cell tumour derived from germ cell neoplasia in situ (GCNIS derived) included one teratoma, six with mixed germ cell tumours and one embryonal carcinoma. Two males had sex cord stromal tumours: (Leydig cell and granulosa cell biology). Twenty-eight patients underwent high radical inguinal orchidectomy(s) with one male also requiring retroperitoneal surgery to clear distant locoregional disease and a further single case thoracotomy and metastasectomy. Six patients had lesions suitable for 'testicular sparing' surgery. Six patients had metastatic disease at presentation (18%). Overall study survival was 97%. A single fatality occurred in an adolescent male with a mixed GCT harbouring liver, lung and para-aortic disease who died 48 months after initiating treatment. CONCLUSION: We highlight one of the largest study series of paediatric testicular tumours in the UK and Europe. Non-GCNIS derived tumours accounted for the most common tumour biology (56%). Survival for paediatric testicular tumours is reassuringly generally excellent. Delayed presentation however with a malignant testicular tumour may be associated with poor outcome(s).

BET inhibition therapy counteracts cancer cell survival, clonogenic potential and radioresistance mechanisms in rhabdomyosarcoma cells.

The antitumour effects of OTX015, a first-in-class BET inhibitor (BETi), were investigated as a single agent or in combination with ionizing radiation (IR) in preclinical in vitro models of rhabdomyosarcoma (RMS), the most common childhood soft tissue sarcoma. Herein, we demonstrated the upregulation of BET Bromodomain gene expression in RMS tumour biopsies and cell lines compared to normal skeletal muscle. In vitro experiments showed that OTX015 significantly reduced RMS cell proliferation by altering cell cycle modulators and apoptotic related proteins due to the accumulation of DNA breaks that cells are unable to repair. Interestingly, OTX015 also impaired migration capacity and tumour-sphere architecture by downregulating pro-stemness genes and was able to potentiate ionizing radiation effects by reducing the expression of different drivers of tumour dissemination and resistance mechanisms, including the GNL3 gene, that we correlated for the first time with the RMS phenotype. In conclusion, our research sheds further light on the molecular events of OTX015 action against RMS cells and indicates this novel BETi as an effective option to improve therapeutic strategies and overcome the development of resistant cancer cells in patients with RMS.

Novel technique for parietal pleural dissection using a laparoscopic hernia balloon.

We describe a novel technique for the creation of a pleural tent and pleurectomy via the use of a laparoscopic hernia balloon. In this method a Spacemaker™ Structural Balloon Trocar (Covidien, USA) is tunnelled under the pleura at the site of thoracotomy or video assisted thoracoscopic surgery port and incrementally inflated under vision. This method is less traumatic than traditional methods, is more likely to provide an intact pleural tent, and allows the surgeon to operate in a near bloodless operative field.

Ameloblastic Fibroma of the Mandible Reconstructed with Autogenous Parietal Bone: Report of a Case and Literature Review.

Ameloblastic fibroma (AF) is a rare, slow-growing benign neoplasm, comprised of tissues of odontogenic origin. It constitutes 2% of odontogenic tumours, occurring at any age, but has a predilection to present in the first two decades of life. AF principally affects the posterior mandible. It is characterized by epithelial islands and cords immersed in ectomesenchyme that mimics the dental papilla and enamel organ but without actual hard tissue formation. Herein, we describe the case of a 6-year-old Caucasian male who presented to the Oral and Maxillofacial Department at Alder Hey Children's Hospital, Liverpool, UK, with a painless expansile mass in the left mandible which was diagnosed as a benign ameloblastic fibroma and subsequently enucleated and reconstructed with a parietal calvarial bone graft. A brief literature review and the issues surrounding diagnosis are discussed.

A study of attributable variables impacting orthopedic trauma surgical training.

PURPOSE: In medical colleges, resident training programs must provide adequate surgical experiences, making them qualified at the end of residency program. It is generally believed that it would take more time for a surgical resident to perform surgical procedures than a board-certified surgeon. There is no current benchmark with regards to operative time and surgical cases involving orthopedic surgery residents. In this study, we focused on two key aspects of surgical training variables, namely, surgical duration & C-arm shoots when the procedure is done by a faculty surgeon in comparison to done by an orthopedic resident under supervision of faculty surgeon. METHODS: It is an observational prospective study, we observed patients undergoing 1 of 5 common orthopedic trauma operations in a community teaching hospital. We recorded two variables, 'skin to skin' surgical duration & number of image intensifier television/C-arm shoots of faculty surgeons and orthopedic resident (postgraduate-3yr) under supervision of faculty surgeon. We calculated mean difference of two variables with or without resident & determined statistical significance, we also compared functional outcome at final follow-up. RESULTS: The total number of procedure observed was 402. On observing summarized results of all surgical procedures, faculty surgeons took on an average 33 min lesser (p < 0.05) & on an average 37 lesser number of shoots (p < 0.05) than resident surgeons. The difference in surgical duration tended to increase with the greater complexity of the surgical dissection. The difference in number of C-arm shoots tended to increase with the increase in surgical duration in closed procedures. In all the five procedures there was no significant difference (p > 0.05) in functional outcome of cases performed by faulty surgeon and resident. CONCLUSION: Little data has been previously published regarding the impact of teaching orthopedic resident in operating room. We demonstrate that resident participation increases the procedure time for commonly performed orthopedic procedures and also the number of C-arm shoots, hence there is a need for technical training facilities outside the operating room such as in cadaveric labs, saw bone labs & virtual surgery simulation. Also the preoperative plan should be thoroughly discussed by faculty surgeon with residents.

Excision of Giant Schwannoma in a Nonagenarian-operative techniques for enhanced recovery after thoracotomy in the high-risk patient.

Thoracotomy is acknowledged as one of the most painful procedures in surgical practice, with the potential to result in significant acute and chronic sequelae, which become especially relevant in high-risk patient populations. Certain pathologies necessitate this surgical approach, and in those circumstances we must aim to mitigate postoperative complications by employing surgical techniques that decrease the risk of nerve injury, rib fracture, and unnecessary soft tissue trauma. We describe an approach to thoracotomy that incorporates evidence-based strategies to lessen the risk of these potential complications, which resulted in rapid postoperative recovery in a nonagenarian.

Hepatoblastoma and Wilms' tumour in an infant with Beckwith-Wiedemann syndrome and diazoxide resistant congenital hyperinsulinism.

Beckwith-Wiedemann syndrome (BWS) can be associated with embryonal tumours and congenital hyperinsulinism (CHI). We present an infant with BWS who developed congenital hepatoblastoma and Wilms' tumour during infancy. The infant presented with recurrent hypoglycaemia requiring high intravenous glucose infusion and was biochemically confirmed to have CHI. He was resistant to diazoxide but responded well to octreotide and was switched to Lanreotide at 1 year of age. Genetic analysis for mutations of ABCC8 and KCNJ11 were negative. He had clinical features suggestive of BWS. Methylation-sensitive multiplex ligation-dependent probe amplification revealed hypomethylation at KCNQ1OT1:TSS-DMR and hypermethylation at H19 /IGF2:IG-DMR consistent with mosaic UPD(11p15). Hepatoblastoma was detected on day 4 of life, which was resistant to chemotherapy, requiring surgical resection. He developed Wilms' tumour at 3 months of age, which also showed poor response to induction chemotherapy with vincristine and actinomycin D. Surgical resection of Wilms' tumour was followed by post-operative chemotherapy intensified with cycles containing cyclophosphamide, doxorubicin, carboplatin and etoposide, in addition to receiving flank radiotherapy. We report, for the first time, an uncommon association of hepatoblastoma and Wilms' tumour in BWS in early infancy. Early onset tumours may show resistance to chemotherapy. UPD(11p15) is likely associated with persistent CHI in BWS. Learning points: Long-acting somatostatin analogues are effective in managing persistent CHI in BWS. UPD(11)pat genotype may be a pointer to persistent and severe CHI. Hepatoblastoma and Wilms' tumour may have an onset within early infancy and early tumour surveillance is essential. Tumours associated with earlier onset may be resistant to recognised first-line chemotherapy.

A Long-Term Study of Application of Joshi's External Stabilizing System in Displaced Intra-articular Distal End Radius Fractures.

Background Fractures of the distal end radius are a common upper extremity fracture. Intra-articular distal end radius fractures are recognized as very complex injuries with a variable prognosis. The aim of the study was to assess the long-term functional outcome of patients treated with Joshi's external stabilizing system (JESS) for displaced intra-articular distal end radius fractures. Materials and Methods A total of 170 patients with intra-articular distal end radius fracture were treated with JESS from 2014 to 2017. The patients were followed up at 2, 6 weeks, 6 months, 1, and 2 years (final) after the surgery. The assessment of pain, range of motion, grip strength, and satisfaction were assessed at 6 months, 1, and 2 years (final) follow-up and scored according to modified Mayo wrist scoring system. Results The good and/or excellent results were found in 82.2% of cases. We observed that patients with age less than 50 years had greater prognosis as compared with patients with more than 50 years of age. Final outcome was also found better in males as compared with females at 6 months, 1, and 2 years postoperatively. Conclusion JESS is an effective treatment technique for intra-articular distal end radius fractures in our community. On long-term follow-up of the patients treated with JESS for intra-articular distal end radius fractures, the functional and radiological outcomes were good with low complication rate.

Aneurysmal Bone Cyst-An Unusual Presentation of Wrist Pain.

Introduction Aneurysmal bone cyst (ABC) is a cystic benign expansile lytic bone lesion, resembling multiple intraosseous blood-filled spaces. It is most commonly seen in the first two decades of life. Most frequent sites of involvement are metaphysis of proximal and distal femur, proximal tibia, and posterior elements of spine. It is seen more frequently in females. ABC of distal ulna is rarely reported in the literature. Case Presentation We are reporting a case of ABC of a rare location-distal ulna-in a young male, which was successfully treated with extended curettage, and the cavity was filled with bone graft, with K-wire stabilization.

PARP inhibitors affect growth, survival and radiation susceptibility of human alveolar and embryonal rhabdomyosarcoma cell lines.

PURPOSE: PARP inhibitors (PARPi) are used in a wide range of human solid tumours but a limited evidence is reported in rhabdomyosarcoma (RMS), the most frequent childhood soft-tissue sarcoma. The cellular and molecular effects of Olaparib, a specific PARP1/2 inhibitor, and AZD2461, a newly synthesized PARP1/2/3 inhibitor, were assessed in alveolar and embryonal RMS cells both as single-agent and in combination with ionizing radiation (IR). METHODS: Cell viability was monitored by trypan blue exclusion dye assays. Cell cycle progression and apoptosis were measured by flow cytometry, and alterations of specific molecular markers were investigated by, Real Time PCR, Western blotting and immunofluorescence experiments. Irradiations were carried out at a dose rate of 2 Gy (190 UM/min) or 4 Gy (380 UM/min). Radiosensitivity was assessed by using clonogenic assays. RESULTS: Olaparib and AZD2461 dose-dependently reduced growth of both RH30 and RD cells by arresting growth at G2/M phase and by modulating the expression, activation and subcellular localization of specific cell cycle regulators. Downregulation of phospho-AKT levels and accumulation of γH2AX, a specific marker of DNA damage, were significantly and persistently induced by Olaparib and AZD2461 exposure, this leading to apoptosis-related cell death. Both PARPi significantly enhanced the effects of IR by accumulating DNA damage, increasing G2 arrest and drastically reducing the clonogenic capacity of RMS-cotreated cells. CONCLUSIONS: This study suggests that the combined exposure to PARPi and IR might display a role in the treatment of RMS tumours compared with single-agent exposure, since stronger cytotoxic effects are induced, and compensatory survival mechanisms are prevented.

Seasonal variation in orthopedic trauma patients-An experience from central India.

OBJECTIVES: This study aims to determine the various epidemiological factors responsible for orthopedic trauma cases: how do weather patterns, month, season and public working schedule influence the daily frequency of orthopedic trauma. MATERIAL AND METHODS: This was a descriptive study performed in the Department of Orthopedics at a tertiary care centre in central India Participants: 7980 trauma cases reported in our study period. Study variables: Demographic characteristics of the cases, time, day, the month of injury and type of trauma and cause of trauma. etc. Statistical analysis: Proportions. RESULTS: In our study period from 2005 to 2016, there were total 7980 admissions, the annual incidence of trauma was 22.78%. RTA was the commonest cause of injury (46.85%). Most common age group affected was 11-40 year age group (64.06%), with the predominance of Male (67.40%) and rural population (72%). The commonest victims of trauma were labourer (37.66%). Maximum cases of trauma occurred during summer (58.9%). Fracture of upper extremity especially around Elbow was common which were 987 (26.41%) amongst which the fracture supracondylar humerus in pediatric age group was most common around 456. And in rainy season and winter season, there was the dominance of lower extremity fracture which was fracture around Ankle and Foot (i.e 557; 23.59%) and fracture of tibia bone (i.e 516; 27.4%) respectively. CONCLUSION: Orthopedic trauma at a tertiary health care trauma center do vary significantly with the weather and are highest in the Summer season.

Minimally Invasive Plate Osteosynthesis Using Locking Plates for AO 43-Type Fractures: Lessons Learnt From a Prospective Study.

INTRODUCTION: Managing fractures of distal tibia is still a subject of debate for orthopaedic surgeons in terms of both, reduction and fixation. Subcutaneous location and soft tissue anatomy predisposes it to angular and rotational instability as well as other bony and soft tissue complexities. Minimally invasive plating offers many advantages over conventional open techniques. It causes minimal soft tissue dissection and surgical trauma to the bone. Minimally invasive plate osteosynthesis(MIPO) maintains biological configuration of distal tibia and fracture hematoma and also provides a construct, which is biomechanically more stable. OBJECTIVES: Evaluation of results of MIPO in management of distal tibia fractures considering radiological union, ankle function restoration, and complications. MATERIALS AND METHODS: In our study, 25 closed distal one-third tibia fracture with/without articular extension were taken, fulfilling the inclusion criteria (AO classification: 10, 43A1; 3, 43A2; 2, 43B1; 4, 43B2; and 6, 43C1). MIPO with locking plates was the treatment undertaken. Patients were followed up for 18 months prospectively. RESULTS: Average injury-hospital interval was 11.16 hours and average injury-operation interval was 2.44 days. All fractures showed radiological union at an average duration of 20.5 weeks (14-28 weeks). Olerud and Molander score was used for evaluation at 3, 6, and 18 months. One patient had union with valgus angulation >5° but there was no nonunion. There was 1 superficial postoperative wound infection. CONCLUSION: Our study shows that plating with MIPO is an effective treatment for closed distal one-third tibia fractures, considering union time and complications rate. Younger age promotes early union and functional recovery. LEVELS OF EVIDENCE: Therapeutic, Level II: Prospective.

Assessing the Effectiveness of a Far-Red Fluorescent Reporter for Tracking Stem Cells In Vivo.

Far-red fluorescent reporter genes can be used for tracking cells non-invasively in vivo using fluorescence imaging. Here, we investigate the effectiveness of the far-red fluorescent protein, E2-Crimson (E2C), for tracking mouse embryonic cells (mESCs) in vivo following subcutaneous administration into mice. Using a knock-in strategy, we introduced E2C into the Rosa26 locus of an E14-Bra-GFP mESC line, and after confirming that the E2C had no obvious effect on the phenotype of the mESCs, we injected them into mice and imaged them over nine days. The results showed that fluorescence intensity was weak, and cells could only be detected when injected at high densities. Furthermore, intensity peaked on day 4 and then started to decrease, despite the fact that tumour volume continued to increase beyond day 4. Histopathological analysis showed that although E2C fluorescence could barely be detected in vivo at day 9, analysis of frozen sections indicated that all mESCs within the tumours continued to express E2C. We hypothesise that the decrease in fluorescence intensity in vivo was probably due to the fact that the mESC tumours became more vascular with time, thus leading to increased absorbance of E2C fluorescence by haemoglobin. We conclude that the E2C reporter has limited use for tracking cells in vivo, at least when introduced as a single copy into the Rosa26 locus.

Pharmacological targeting of the ephrin receptor kinase signalling by GLPG1790 in vitro and in vivo reverts oncophenotype, induces myogenic differentiation and radiosensitizes embryonal rhabdomyosarcoma cells.

BACKGROUND: EPH (erythropoietin-producing hepatocellular) receptors are clinically relevant targets in several malignancies. This report describes the effects of GLPG1790, a new potent pan-EPH inhibitor, in human embryonal rhabdomyosarcoma (ERMS) cell lines. METHODS: EPH-A2 and Ephrin-A1 mRNA expression was quantified by real-time PCR in 14 ERMS tumour samples and in normal skeletal muscle (NSM). GLPG1790 effects were tested in RD and TE671 cell lines, two in vitro models of ERMS, by performing flow cytometry analysis, Western blotting and immunofluorescence experiments. RNA interfering experiments were performed to assess the role of specific EPH receptors. Radiations were delivered using an x-6 MV photon linear accelerator. GLPG1790 (30 mg/kg) in vivo activity alone or in combination with irradiation (2 Gy) was determined in murine xenografts. RESULTS: Our study showed, for the first time, a significant upregulation of EPH-A2 receptor and Ephrin-A1 ligand in ERMS primary biopsies in comparison to NSM. GLPG1790 in vitro induced G1-growth arrest as demonstrated by Rb, Cyclin A and Cyclin B1 decrease, as well as by p21 and p27 increment. GLPG1790 reduced migratory capacity and clonogenic potential of ERMS cells, prevented rhabdosphere formation and downregulated CD133, CXCR4 and Nanog stem cell markers. Drug treatment committed ERMS cells towards skeletal muscle differentiation by inducing a myogenic-like phenotype and increasing MYOD1, Myogenin and MyHC levels. Furthermore, GLPG1790 significantly radiosensitized ERMS cells by impairing the DNA double-strand break repair pathway. Silencing of both EPH-A2 and EPH-B2, two receptors preferentially targeted by GLPG1790, closely matched the effects of the EPH pharmacological inhibition. GLPG1790 and radiation combined treatments reduced tumour mass by 83% in mouse TE671 xenografts. CONCLUSIONS: Taken together, our data suggest that altered EPH signalling plays a key role in ERMS development and that its pharmacological inhibition might represent a potential therapeutic strategy to impair stemness and to rescue myogenic program in ERMS cells.