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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.
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Objective To clone silent information regulator 1 (SIRT1) gene full-length cDNA,construct recombinant eukaryotic expression vector containing SIRT1 gene and its mutant T200I,E420K,so as to lay the foundation for further research of SIRT1 gene function.Methods RT-PCR amplified SIRT1 gene full-length cDNA.PCR products were cloned into the eukaryotic expression vector pcDNA3.1 (+) through double digestion and pcDNA3.1(+)-SIRT1 recombinant plasmid was obtained.Meanwhile,site-directed mutagenesis was applied to build its mutant pcDNA3.1 (+)-T200I and pcDNA3.1 (+)-E420K expression vector.Recombinant plasmid was identified by enzyme digestion and DNA sequencing and the recombinant eukaryotic expression vector of success was screened out.Results SIRT1 gene full-length cDNA was successfully cloned,and pcDNA3.1 (+)-SIRT1 eukaryotic expression vector and its mutant were also successfully constructed.Positive recombinant plasmid sequencing was compared after enzyme digestion,and it was completely consistent with the expected sequence.Transfected 293T cell line was established and HIS tagged SIRT1 protein was expressed.Conclusions We successfully constructed pcDNA3.1 (+)-SIRT1 and its mutant pcDNA3.1 (+)-T200I,pcDNA3.1 (+)-E420K eukaryotic expression vector,which may provide genetic material for biological function study of SIRT1 gene and its mutant T200I,E420K.
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ObjectiveTo investigate the continuous pathological features of biopsy specimens from five cases of small bowel allotransplantation (SBT) in order to provide more reliable information for the diagnosis and treatment of acute rejection (AR) in SBT.Methods324 biopsy specimens of intestinal mucosa after SBT from 5 patients were collected and studied by histology,histochemistry and electron microscopy.ResultsIn the early stage after operation (0～3 months),AR IND-1 grade was diagnosed for four times on 3 of 5 patients.During 3-6 months,AR IND-1 grade for three times was diagnosed in 2 cases,and AR 2 grade for two times during 7 ～ 12 months. All the patients suffered ischemia reperfusion injury, lymphatic vessel reconstruction and AR.Conclusion The pathological examination of biopsy specimens of intestinal mucosa is still the most reliable detecting method to diagnose AR,and continuous observation may play an important role to monitor the occurrence,development,and treatment response of AR. The final diagnosis of AR depends on structure of intestinal mucosa,crypt epithelium injury and inflammatory cells infiltration. The communication among the pathologist and surgeon is the best way to reduce misdiagnoses.Ultrastructural examination is used to verify the pathogenic microorganism.
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Objective To investigate the clinical presentation, endoscopy and pathological features of subclinical cellular rejection (SCR) of small bowel allotransplantation. Methods Three times of SCR in a patient after isolated small bowel transplantation were studied by endoscopy and microscopy, and the clinical data and literature were reviewed. Results SCR was an unusual type of acute rejection after small bowel transplantation. SCR showed low-grade morphological changes of acute rejection, and may be relived after low-dose steroid or bolus steroid was given. Conclusion The causes of SCR are not clear now. SCR may be the early stage of clinical acute rejections, and may be correlated with unexpected high grade acute rejection, and chronic loss function of graft. The biopsy through ileoscopy is a "golden standard" of diagnosis of SCR in small bowel transplantation.However, the vessel lesions of graft, ileus, and inflammation should be excluded before diagnosis.
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Objective To explore the morphological, immunohistochemical characters and prognosis in one case of patients with cutaneous anaplastic large cell lymphoma complicated with acute myeloid leukemia (C-ALCL-AML). Methods The histopathology, immunohistochemical markers and follow-up information of one case of ALCL-AML was analyzed and the correlated literature was reviewed. Results The patient, 69 year-old, female, was initially present with shin lesion on one finger and abnormal myelogram. The histopathology of shin lesion showed that tumor cells were composed of large cells with abundant cytoplasm,the nuclei were large and irregular, and were infiltrated by Neutrophil and eosinophil. The CD30,CD3 and CD43 of tumor cells were positive, but ALK negative by immunohistochemical method. The number of WBC in peripheral blood was 15.5×109/L and 51 archaeocytes were in every 100 karyotes. Bone marrow aspiration detection showed that bone marrow was hyperplasia and the ratio of myeloblast was 78 %. This patient was diagnosed as C-ALCL-AML, partly differentiation type(AML-M2a). Conclusion C-ALCL-AML is very rare. Its diagnosis is dependent on clinical data, histopathology and immunohistochemical markers. The first choice of treatment is chemotherapy, but its prognosis is poor.
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Objective To observe the effects of mouse indoleamine 2,3-dioxygenase (IDO) gene on CD4+ T cells by transfecting an eukaryotic expression vector containing mouse IDO gene fused with enhanced green fluorescent protein (pEGFP-N1). Methods The immature dendritic cells (imDCs) derived from mouse bone marrow were cultured in vitro and morphologically identified by transmission electron microscope, scanning electron microscope and flow cytometry. The immature dendritic cells were transfected by DOTAP liposome, and the expression of green fluorescence protein was confirmed with inverted fluorescence microscope. The spleen-derived CD4+ T lymphocytes from mice were isolated and purified in vitro, and then carried to mixed lymphocyte reaction (MLR) with 3H-TdR incorporation. The effect of IDO gene on the proliferation of spleen-derived CD4+ T lymphocytes of the allergized mouse challenged by ovalbumin was assayed by 3H-TdR MLR incorporation, and the apoptosis of spleen-derived CD4+ T lymphocytes was assayed by TUNEL. Results The proliferation of spleen-derived CD4+ T lymphocytes of the allergized mouse challenged by ovalbumin were inhibited by the imDCs transfected with IDO gene, the number of CD4+ T lymphocytes treated with the imDCs transfected pEGFP-N1-IDO eukaryotic expression vector decreased significantly compared with that in control group (P