FLI1 gene influences lesion size and skin test may predict therapeutic response in cutaneous leishmaniasis

Genes associated with wound healing have been shown to be risk factors for cutaneous leishmaniasis (CL) which is caused by Leishmania braziliensis. In this study, we examined whether the genes previously associated with CL influenced the clinical outcome. Patients were genotyped and retrospectively classified as responders, who were cured with a single course of pentavalent antimony (Sbv), or as refractories, who did not respond to Sbv. Patients characterised as responders showed a stronger response to the leishmanin skin test (LST) when compared to the refractory subjects (p = 0.0003). Furthermore, we observed an association between the FLI1 CC genotype and an increased size of ulcers (p = 0.0170). We suggest that the leishmanin skin test may be a predictive tool for therapeutic outcome and reinforce FLI1 as a potential influencer of susceptibility and lesion size in CL.

FLI1 and MIC2 expression in precursor B-lymphoblastic leukemia with Burkitt-like morphology and extensive extramedullary involvement: A diagnostic challenge in pediatric small round cell tumor

Pediatric small round cell tumors (PSRCTs) constitute a large proportion of childhood malignancies with overlapping diagnostic and clinical features but radically different therapies. Here, we report a case of 16-year-old male child presenting with diffuse abdominal and mediastinal mass, axillary lymphadenopathy, and pleural effusion. Bone marrow aspirate showed near total replacement by small round malignant cells. The bone marrow biopsy showed interstitial infiltration by malignant cells, which were CD45? CD3? CD20? MIC2+ FLI1+ and diagnosis of Ewing's sarcoma was established. In contrast, flowcytometric immunophenotyping of the bone marrow aspirate showed CD45? cells, which were CD19+ cytCD79a+ CD10+ CD81+ CD38+ HLA-DR+ CD22+ CD20? consistent with B-cell acute lymphoblastic leukemia (B-ALL). The extended immunostaining panel on bone marrow biopsy also showed positivity for cytCD79a, CD10, CD19, and BCL-2, whereas fluorescent in-situ hybridization for EWSR1 gene rearrangement was negative. Thus, a final diagnosis of CD45? FLI1+ MIC2+ B-ALL was established. Rare cases of CD45? B-ALL with immunoreactivity for MIC2 and Friend leukemia virus integration 1 (FLI1) have posed a diagnostic challenge for PSRCTs in the recent past. This case report highlights the role of multimodality approach in establishing a correct diagnosis in CD45? PSRCTs to ensure definitive therapy and better clinical outcome.

Ewing Sarcoma / 임상소아혈액종양

Ewing sarcoma is the second most frequently occurring malignant tumor of the bone and soft tissue in adolescents and young adults. Genetically, Ewing sarcoma is characterized by balanced chromosomal translocation in which a member of FET gene family is fused with an ETS transcription factor, with the most common fusion being EWSR1-FLI1 (85% of cases). Treatment of Ewing sarcoma is based on multidisciplinary approach (local surgery, radiotherapy and multiagent chemotherapy), which are associated with chronic late effects that may compromise quality of life of survivors. First line treatment includes combination of drugs incorporating doxorubicin, vincristine, cyclophosphamide, ifosfamide, etoposide, and dactinomycin. The beneficial role of high dose chemotherapy has been suggested in high-risk localized Ewing sarcoma patients, and the studies are being performed to investigate the role in metastatic disease. The 5-year overall survival for localized Ewing sarcoma has improved to reach 65% to 75%. But patients with metastatic disease have a 5-year survival rate of <30%, except for those with isolated pulmonary metastasis (approximately 50%). Patients with recurrent tumor have a dismal prognosis. Novel therapeutic strategies based on understanding of molecular mechanisms are needed to improve the outcome of Ewing sarcoma and to lessen the treatment-related late effects.

Effects of CXCL4 on human dermal microvascular endothelial cells and secretion of vasomotion factors / 中国免疫学杂志

Objective:To identify the proliferation effect and angiogenic ability of CXCL4 on human dermal microvascular endothelial cells(HDMECs),and to explore the secretion of vasomotion factors.Methods:HDMECs were treated with gradient concentration to test the proliferation of HDMECs.CCK-8 was used to explicated the proliferation of HDMECs.The effect of CXCL4 on angiogenic ability of HDMECs was determined by tube formation assay.The mRNA levels of endothelin-1(ET-1),Fli-1,AngiotensinⅡtype 1 receptor(AT1R) and endothelin-1 type A receptor (ETAR) were detected by real-time quantitative polymerase chain reaction (Real-time PCR).Results:The specific receptor of CXCL4 was expressed on HDMECs.CXCL4 could inhibit the proliferation of HDMECs and the number of tube formation in a dose-depend manner.After CXCL4 intervention,the relative amplification multiples of ET-1,AT1R were significantly increased(P<0.05),Fli-1 was decreased(P<0.05),and ETAR had no change as compared with the control group.Furthermore,CXCL4 antagonist could reverse the effects of CXCL4 on HDMECs.Conclusion:CXCL4 inhibit the proliferation and angiogenesis of HDMECs and induce the secretion of ET-1 and AT1R,reduce the secretion of Fli-1 in a dose-dependent manner.

Characteristics of ERG, Fli-1, CD34, CD31 and FⅧRAg expression in hepatic malignant vascular tumors / 中华病理学杂志

Objective@#To investigate the expression of ERG, Fli-1, CD34, CD31 and factor Ⅷ-related antigen(FⅧRAg) in hepatic malignant vascular tumors.@*Methods@#A retrospective analysis was conducted on 63 cases of primary hepatic malignant vascular tumors and 31 cases of hepatic other malignant spindle cell tumors collected during January 1986 to January 2014. EnVision method was used to detect the expression of ERG, Fli-1, CD34, CD31, FⅧRAg.@*Results@#Sixty-three cases of malignant vascular tumors, including 24 cases of angiosarcoma, 38 cases of epithelioid hemangioendothelioma and 1 case of hepatic Kaposi′s sarcoma. All of the cases were positive for ERG(100.0%, 63/63). Positive rate of Fli-1, CD34, CD31, FⅧRAg was 96.8% (61/63), 87.3% (55/63), 81.0% (51/63) and 41.3% (26/63), respectively. In other hepatic malignant spindle cell tumors, the positive rate of ERG, Fli-1, CD34, CD31 and FⅧRAg was 3.2% (1/31), 19.4% (6/31), 19.4% (6/31), 9.7%(3/31) and 3.2%(1/31), respectively.The sensitivity of ERG, Fli-1, CD34, CD31, FⅧRAg was 100.0%, 96.8%, 87.3%, 81.0% and 41.3%, respectively.The specificity was 96.8%, 80.6%, 80.6%, 90.3% and 96.8%, respectively.@*Conclusion@#ERG is a more sensitive and specific diagnostic marker for hepatic malignant vascular tumors in comparison to Fli-1, CD34, CD31 and FⅧRAg.

Expression of FLi-1, h-caldesmon and collagen Ⅳ in glomus tumor: a clinicopathologic study / 临床与实验病理学杂志

Purpose To explore the expression of FLi-1,h-caldesmon and collagen Ⅳ in glomus tumor and possible relationship between different histological types and clinical-pathological characteristics.Methods Immunohistochemistry of EnVision method was used to detect the expression of FLi-1,h-caldesmon and collagen Ⅳ in 35 cases of glomus tumor under microscope,and the relationship between different histological types and clinical-pathological characteristics was analyzed.Results The pathological morphology of tumor cells was round or polygonal,patchy distribution in blood vessels or arranged in a ring around the blood vessels.The tumor cell boundary was clear and the shape was regular,gradual transition between tumor cells and spindle smooth muscle cells was sometimes visible.The positive rate of FLi-1,h-caldesmon and collagen Ⅳ in 35 cases of glomus tumor were 58.6％,97.1％ and 100％ respectively.Different histological types of glomus tumors had no correlation with the sex of patient and the size of tumor,but had correlation with the age of patient.Vimentin and SMA were positive for glomus tumors by immunohistochemistry in 35 cases.CD34 was positive in 2 cases.Desmin positive was found in 1 case.EMA,S-100,CgA,CD68 and CD99 were negative.Conclusion h-caldesmon and collagen Ⅳ are the markers for glomus tumor diagnosis,and FLi-1 can serve as a good auxiliary reference marker.

A rare case of multifocal pseudomyogenic hemangioendothelioma, involving soft tissues and bone, misdiagnosed as a rhabdomyosarcoma: Diagnostic and treatment implications.

Pseudomyogenic hemangioendothelioma (PHE) is an uncommon, but distinctive soft tissue tumor, characterized by multifocality. A 17‑year‑old male referred to us with progressively increasing multiple subcutaneous nodular lesions over his left leg and foot, reported elsewhere as a spindle cell rhabdomyosarcoma. On review, microscopy showed a cellular tumor comprising plump spindle cells arranged in loose fascicles with interspersed inflammatory cells. Tumor cells exhibited mild nuclear variation. Immunohistochemically, tumor cells expressed AE1/AE3, CD31, Fli‑1, and smooth muscle actin (SMA), confirming diagnosis of PHE. Whole‑body positron emission tomography–computed tomography (PET‑CT) scan revealed multiple, metabolically active, subcutaneous nodular lesions over the left lower leg and in the distal tibia. Subsequently, resection specimens from the various lesions and bone curettage also revealed features of PHE. Three months later, the patient developed multiple lesions over his fourth toe and left foot, for which he underwent tumor resections. At present, he is disease‑free. PHE is a locally aggressive soft tissue tumor characterized by multifocality, rarely bony involvement and can be misdiagnosed as a high‑grade sarcoma.

Primary primitive neuroectodermal tumor of kidney: A rare case report with diagnostic challenge.

Primary primitive neuroectodermal tumors (PNETs) of the kidney are quite rare and can be mistaken for a wide variety of other small round blue cell tumors which includes rhabdomyosarcoma, Wilm’s tumor, carcinoid, neuroblastoma, clear cell sarcoma of the kidney, lymphoma etc. Renal Ewings/PNET can occur in the age group from 4 to 61 years. Approximately, 90% of Ewing sarcoma (ES)/ PNET have a specifi c t(11;22) which results in a chimeric EWS-FLI-1 fusion protein. Immunohistochemical for the carboxy-terminus of FLI-1 is sensitive and highly specifi c for the diagnosis of ES/PNET. Herein, we have an interesting presentation in a 23-year-old male who came with neck pain and progressive quadriparesis and was diagnosed as a case of poorly differentiated malignant tumor with a differential of lymphoma versus metastatic renal cell carcinoma. The patient’s condition deteriorated fast and he had a rapid downhill course. The fi nal diagnosis of Ewings/PNET was confi rmed at autopsy.

The construction and identification of prokaryotic expression vector pQE30-EWS-FLI1 / 重庆医科大学学报

Objective:To construct Ewing's sarcoma EWS-FLI1 gene prokaryotic expression vector.Methods:The target gene of EWS-FLI1 was obtained by RT-PCR method after the total RNA was extracted from Ewing's sarcoma A673 cells.The site sequences of restrictive endonuclease SacⅠand Hind Ⅲ were introduced into the upstream and downstream of target gene respectively.The target gene fragment were cloned into pMD18-T and transformed into E.Coli JM109.Screened positive clones were confirmed by PCR,restrictive endonuclease digestion and DNA sequencing.The EWS-FLI1 gene was sequentially subcloned into prokaryotic expression vector pQE30,and the recombinant plasmid pQE30-EWS-FLI1 was confirmed by restrictive endonuclease digestion and DNA sequencing.The proteins,expressed in E.coli JM109 transformed with EWS-FLI1recombinant plasmid under IPTG induction,were characterized by SDS-PAGE and Western-blot.Results:PCR result indicated that an amplified DNA fragment was in size of 1.5 kb.Restrictive endonuclease digestion analysis indicated that the target gene was in size of 1.5 kb.DNA sequencing analysis demonstrated that sequence of target gene accorded with anticipated one.The EWS-FLI1 with a molecular weight of 54 kD was highly expressed in pQE30-EWS-FLI1.Western blot proved that the expressed product had the antigenicity of EWS-FLI1.Conclusion:The recombinant prokaryotic expression vector pQE30EWS-FLI1 is constructed successfully,which will contribute to the further research of EWS-FLI1.