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Journal of Zhejiang University. Medical sciences ; (6): 193-199, 2014.
Article in Chinese | WPRIM | ID: wpr-336719


<p><b>OBJECTIVE</b>To construct and identify lentiviral vector containing human ILK-shRNA and mda7 gene.</p><p><b>METHODS</b>Based on the human ILK gene sequences, RNAi target sequences were designed and cloned into the lentiviral vector pSicoR-eGFP by restriction endonuclease HpaI and XhoI double digestion and T4 DNA ligase ligation. Based on the human mda7 gene sequences, PCR primers were designed to clone the full-length mda7, and were cloned into the lentiviral vector pLVX-Puro. After the candidate clones were identified by DNA sequencing, the recombinant plasmid and the three packaging plasmids were co-transfected into the human embryonic kidney 293T cells by lipofectamine 2000 to produce the lentiviral particles. Human prostate cancer PC-3 cells were infected with the constructed lentiviral vector. The ILK and mda7 expression levels in PC-3 cells were quantified by qPCR and Western blot, respectively. The effect of ILK and mda7 on proliferation and migration of PC-3 cells were assessed by MTT method and Transwell assay, respectively.</p><p><b>RESULTS</b>ILK-pSicoR-eGFP and mda7-pLVX-Puro lentiviral vectors were successfully constructed. Strong green fluorescence was observed in the 293T cells under the fluorescent microscope after co-transfection of 293T cells with 4 plasmids of lentiviral vector. The transfection efficiency of the collected virus exceeded 90% in the 293T cells and the PC-3 cells were infected with the lentiviral particles with high efficiency. The A and B lentiviral vector inhibited the expression of ILK at both the mRNA and protein levels in PC-3 cells significantly. The mda7-pLVX-Puro lentiviral vector increased the expression of mda7 in PC-3 cells, and the ability was maintained for one month. Within 96 h, ILK and mad7 significantly inhibited the proliferation and migration of PC-3 cells (Ps<0.05).</p><p><b>CONCLUSION</b>The lentiviral vectors of ILK knockdown and mda7 over-expression have been successfully constructed and identified. The recombinant lentivirus can efficiently infect human prostate cancer PC-3 cells, in which ILK expression is inhibited and mda7 is over-expressed.</p>

Humans , Cell Line , Genetic Vectors , Interleukins , Genetics , Lentivirus , Genetics , Plasmids , Genetics , Protein Serine-Threonine Kinases , Genetics , RNA, Small Interfering , Genetics , Transfection
Chinese Journal of Pathology ; (12): 91-94, 2009.
Article in Chinese | WPRIM | ID: wpr-319780


<p><b>OBJECTIVE</b>To characterize the risks and histopathological features of parvovirus B19 infection of infants in perinatal period.</p><p><b>METHODS</b>Routine pathological examination was performed on 1 neonate, 2 dead fetuses and 2 placentas using either autopsy or biopsy materials.</p><p><b>RESULTS</b>The diagnostic intranuclear inclusions were found in erythroblasts in the bone marrow, liver, spleen and lungs in one case, in the spleen and liver in one case, in the spleen in one case, and in the placentas in two cases.</p><p><b>CONCLUSIONS</b>Severe hemolytic anemia or fetal hydrop or hemophagocytosis caused by the infection of parvovirus B19 can lead to death of infected neonates and fetus. Pathological confirmation of parvovirus B19 infection relies on the identification of erythroblasts containing the diagnostic intranuclear inclusions.</p>

Female , Humans , Infant, Newborn , Pregnancy , Anemia , Pathology , Virology , Autopsy , Biopsy , Erythema Infectiosum , Blood , Pathology , Virology , Erythroblasts , Fetal Death , Fetus , Hydrops Fetalis , Pathology , Virology , Inclusion Bodies , Lymphohistiocytosis, Hemophagocytic , Pathology , Virology , Parvovirus B19, Human , Placenta , Pathology , Virology , Stillbirth
Chinese Journal of Pathology ; (12): 302-306, 2007.
Article in Chinese | WPRIM | ID: wpr-333897


<p><b>OBJECTIVE</b>To study the pathologic features, diagnosis, differential diagnosis and biologic behavior of uterine perivascular epithelioid tumor.</p><p><b>METHODS</b>Five cases of uterine perivascular epithelioid cell tumor were studied by light microscopy and immunohistochemistry. Follow-up information was reviewed.</p><p><b>RESULTS</b>All the five tumors were composed by clear or eosinophilic cells arranged in nests and cords, associated with abundant small vessels and hyalinization in the stroma. Immunohistochemically, the tumor cells demonstrated positive staining for melanocytic markers (HMB45 and/or Melan-A), desmin and smooth muscle actin. The staining for cytokeratin and CD10 was negative. All the patients followed for a certain period are still alive, with no evidence of disease recurrence.</p><p><b>CONCLUSIONS</b>Perivascular epithelioid cell tumor is a rare mesenchymal tumor of uterus, with distinctive histologic and immunohistochemical features. It should be distinguished from clear cell carcinoma and epithelioid leiomyoma of uterus. Positivity for melanocytic markers (especially HMB45) plays an important role in the diagnosis of this tumor. In general, the tumor is categorized as benign, with uncertain malignant potential and malignant.</p>

Adult , Female , Humans , Middle Aged , Adenocarcinoma, Clear Cell , Pathology , Antigens, Neoplasm , Metabolism , Desmin , Metabolism , Diagnosis, Differential , Follow-Up Studies , Hysterectomy , Methods , Immunohistochemistry , Leiomyoma, Epithelioid , Pathology , Melanoma-Specific Antigens , Neoplasm Proteins , Metabolism , Perivascular Epithelioid Cell Neoplasms , Metabolism , Pathology , General Surgery , Uterine Neoplasms , Metabolism , Pathology , General Surgery
Chinese Journal of Hematology ; (12): 583-587, 2004.
Article in Chinese | WPRIM | ID: wpr-229924


<p><b>OBJECTIVE</b>To investigate the rate of dual rearrangements of lymphocytic antigen receptor genes in non-Hodgkin lymphomas (NHL) and its pathogenesis and pathologic significance.</p><p><b>METHODS</b>PCR analysis of monoclonal, polyclonal and dual rearrangements of IgH and TCR gamma, TCR beta genes was carried out in 125 cases of NHL to evaluate the rate of dual rearrangements, immunohistochemistry was performed for a Ki67 protein expression in 117 cases and the proliferation index was calculated. The relationship between antigen receptor gene rearrangements and proliferation index was analyzed.</p><p><b>RESULTS</b>Combination of the two pairs of IgH gene primers with the multiplex PCR for TCR gamma and TCR beta gene revealed dual rearrangements in 8% (8/96) of B-NHL, 17% (5/29) of T-NHL. In B cell NHL, IgH gene monoclonal, dural and polyclonal rearrangements were identified in 65, 8 and 15 cases respectively, while in T-cell NHL, they were in 15, 5 and 9 cases, respectively. There was no significant difference between proliferation index and monoclonal, dual, polyclonal rearrangements in both B-NHL and T-NHL by One-way test.</p><p><b>CONCLUSION</b>Dual rearrangements in NHL are not rare and have no relationship with proliferation index.</p>

Adolescent , Adult , Aged , Aged, 80 and over , Child , Female , Humans , Male , Middle Aged , Young Adult , Carrier Proteins , Genetics , Cell Proliferation , Gene Rearrangement , Genes, T-Cell Receptor gamma , Genetics , Immunohistochemistry , Ki-67 Antigen , Metabolism , Lymphoma, B-Cell , Genetics , Metabolism , Pathology , Lymphoma, Non-Hodgkin , Genetics , Metabolism , Pathology , Lymphoma, T-Cell , Genetics , Metabolism , Pathology , Polymerase Chain Reaction , Receptors, Antigen , Genetics