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Objective@#To investigate the immunohistochemical staining of anaplastic lymphoma kinase (ALK; clone 1A4) in pediatric medulloblastoma (MB).@*Methods@#Molecular subtyping was performed based on the NanoString and sequencing techniques for 44 pediatric MB cases at Children′s Hospital, Zhejiang University School of Medicine from 2014 to 2017. ALK expression was detected with EnVision immunhistochemistry using ALK clone 1A4 on whole section. Statistical analyses were performed to evaluate the correlation of protein expression with molecular subgroups.@*Results@#The age ranged from 0.5 to 13.0 years with an average age of 5.8 years. There were 28 males and 16 females, and 31 classic, 5 desmoplastic nodular, 3 extensive nodular and 5 large cell/anaplastic MBs. Except three cases was unable classified, 41 MBs were classified into the four molecular groups: 5 in WNT group, 12 in SHH group, 9 in Group 3 and 15 in Group 4. Thirteen of 44 MB cases were positive staining for ALK, and the positive rate was 29.5%. Six cases were strong reaction, and 7 cases were weak. The expression of ALK at the protein level was associated with the WNT group (P<0.01). The characteristic perinuclear dot-like staining was only showed in WNT group.@*Conclusions@#The ALK immunhistochemistry using antibody clone 1A4 is a useful marker for the molecular subgroup detection of MB. The strong staining and perinuclear dot-like staining indicate as WNT group.
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<p><b>BACKGROUND</b>The common pathological characteristics of corneal injury include inflammatory factors activation, vascular endothelial cells or inflammatory cells infiltration into lesions, corneal edema, corneal neovascularization (CNV), and scar formation. PEDF-34 is the functional fragment of pigment epithelium-derived factor (PEDF) that has anti-angiogenic and anti-inflammatory properties and contains an N-terminal 34-amino acid peptide. This study was to investigate the anti-inflammatory effects of PEDF-34 on H2O2-induced corneal injury in vitro.</p><p><b>METHODS</b>After cultured in H2O2 (0.1 mmol/L) for 2 hours, human corneal fibroblasts (HCFs) and human umbilical vein endothelial cells (HUVECs) were treated with PEDF-34-nanoparticles (NPs) at different concentrations (0.1, 0.5, 1.0, 2.0 µg/ml) or 2.0 µg/ml control-NPs for 24 hours. The viable cells were quantified using the MTT assay. Western blotting or ELISA analysis was performed for measuring the human vascular endothelial growth factor (VEGF) and intercellular adhesion molecule-1 (ICAM-1) expression of both HCFs and HUVECs. VEGF and nuclear factor κB (NF-κB) mRNA levels of HCFs were semi-quantified by RT-PCR.</p><p><b>RESULTS</b>The survival rates of HCFs or HUVECs stimulated by H2O2 did not decrease significantly (P > 0.05) compared to those in the normal conditions. As compared to control-NP group, PEDF-34-NPs had dose-dependent inhibitive effect on HUVECs with the MTT assay, but not HCFs. Western blotting analysis showed that the VEGF and ICAM-1 levels in the HCFs and HUVECs stimulated by H2O2 were significantly higher than those in the normal conditions, which were decreased dramatically in those treated with PEDF-34-NPs. RT-PCR analysis revealed that the VEGF mRNA and NF-κB mRNA levels increased in H2O2-stimulated HCFs, while both of them decreased in PEDF-34-NP groups dose dependently.</p><p><b>CONCLUSIONS</b>PEDF-34-NPs may play an important role in regulating the NF-κB pathway, inhibiting inflammatory activity. PEDF-34-NPs may be a potential new drug for treating corneal injury in the future.</p>
Subject(s)
Humans , Anti-Inflammatory Agents , Chemistry , Pharmacology , Cells, Cultured , Corneal Injuries , Metabolism , Eye Proteins , Chemistry , Human Umbilical Vein Endothelial Cells , Metabolism , Hydrogen Peroxide , Pharmacology , Nerve Growth Factors , Chemistry , Peptides , Chemistry , Pharmacology , Serpins , ChemistryABSTRACT
Objective To investigate the role of the expression of latent transforming growth factor beta binding protein-1 (LTBP-1) and transforming growth factor beta receptor type Ⅱ(TGF-beta R Ⅱ) in the pathogenesis of condyloma acuminatum (CA). Methods Samples were resected from the lesions of 30 patients with CA and prepuces of 17 normal human controls. The mRNA and protein expressions of LTBP-1 and TGF-betaR Ⅱ were assessed by quantitative real-time PCR and a streptavidin-biotin-peroxidase staining technique, respectively. Results As shown by Real time PCR, the mRNA expression levels of LTBP-1 and TGF-betaR Ⅱ were significantly higher in CA tissues than those in the controls, with the average value of 2 (-Delta Delta α) being 2.46 and 3.43, respectively. A lower intensity of stainning was observed for LTBP-1 and TGF-betaR Ⅱ in CA tissues compared with the normal controls (182.51±9.89 vs 167.78±12.56, 187.35± 11.23 vs 170.15±13.21, t = 5.62, 3.70 respectively, both P <0.01). Conclusion The decrease in the expres-sion of both LTBP-1 and TGF-betaR Ⅱ may lead to the abnormality in the activation of TGF-beta and signal transduction pathways.
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0.05), but not in other brain regions. The number of GFAP-immunopositive cells of the E.coli-treated pups was markedly increased in periventricular white matter and hippocampus at P7 compared with the control group (P0.05). CONCLUSION: Intrauterine infection induces an increased expression of GFAP in the neonatal brain. [
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Objective To evaluate the diagnosis and treatment of intestinal neuronal dysplasia type B (IND) in childhood. MethodsForty-five patients underwent preoperative barium enema examination, 23 patients underwent electromanometry, and mucosal biopsy and immunohistochemical staining for S100 protein were performed in 17 cases. All 45 patients underwent resection of the invalid segment with coloproctostomy. Whole layer was sampled on several sites of the resected segment and examined by two independent pathologists. All patients were followed up from 3 months to 9 years (mean 4.6 years).ResultsTwenty eight patients were complicated with Hirschsprung′s disease, one patient with hypogangliosis, and isolated IND was diagnosed in the other 16 children. The narrowed distal segment with proximal dilatation was merely noted in 4 children with isolated IND. Internal sphincter relaxations were missing in 6 children with isolated IND. The indicative diagnosis might be merely gained in 7 patients by the mucosa biopsy. The correct diagnosis can be established by whole layer biopsy of the resected segment. Three children with enterocolitis after operation were cured by conservative treatment. One patient suffering from postoperative sluice syndrome underwent second resection. Postoperative continence was achived in all patients. ConclusionThe correct diagnosis of IND can be obtained by biopsy of whole layer, and resection of invalid bowel segment with coloproctostomy is the choice of therapy.