Bone Neoplasms ; immunology ; pathology ; Child ; Humans ; Osteosarcoma ; immunology ; pathology ; Rosette Formation

Objective To study the impact of viral infection on stress protein gene expression and investigate the possibility that heat shock proteins interact with viral components during viral assembly in the brain of suckling mice experimentally infected with Hantaan virus (HV). Methods New born mice less than 72 h of age were inoculated into brain with the 100 LD50 of Hantaan virus (strain 76 118). At the 8th day mice died from brain disease. Brains were prepared for tissue sections and extracts for the experiments. The Hantaan virus infection, HSP70 induction and their co localization in brain of infected suckling mice were demonstrated by immunohistochemical staining and laser scan co focal microscopic double labling. The possible intereaction between HV structural proteins and HSP70 was further analyzed by immunoprecipitation of infected tissue extracts with antibodies to either HSP70 or to viral nuclear capsule proteins HV NP by using SDS PAGE and Western blotting. Results HV infection induced an increased levels of HSP70 expression at the cytoplasma of neurons in the brain of suckling mice HV NP protein proved to be co localized with HSP70. Monoclonal antibody (mAb) to HSP70 could co immunoprecipitate the HV NP, and similarly mAb to HV NP could co immunoprecipitate HSP70 from virus infected tissue extracts, but not from mock infected tissue extracts. Antibody, which recognized one of these two proteins, did not cross react with the other. Thus, the association between HSP70 and HV NP appeared to be specific.Conclusions Hantavirus infection can directly induce HSP70 expression, leading to the formation of HV HSP 70 viral antigen peptide complex which plays some roles in the course of viral infection and replication.

Objective To study Heat shock proteins (HSPs) induction and their association with Hantaan virus (HTNV) structural proteins in the brain tissues of mice experimentally infected with HTNV 76～118. Methods Newborn mice less than 72 h of age were experimentally infected with HTNV by intracerebral inoculation. The brains of mice at the 8th day post infection were removed and prepared for tissue extracts. The possible interaction or association of Hantaan virus nucleocapsid protein (HTNV NP) and glycoprotein G2 (HTNV G2) with GRP94, HSP70 and HSP27 were analyzed by double specific antibo dies sandwich ELISA and immunoprecipitation methods. Results HTNV infection induced the expression a set of HSPs including GRP94 and HSP70 in the brain tissues of mice experimentally infected with HTNV. HTNV NP was associated with GRP94, HSP70 and HSP27, which led to formation of HTNV NP HSP70 GRP94 HSP27 complexes. It was also observed that HTNV G2 was associated with NP and HSP27, which led to formation of HTNV G2 NP HSP27 complexes. Conclusions HTNV infection induces expression of a set of HSPs, which were associate with viral structural proteins in the brain tissues of mice experimentally infected with HTNV. This association may demonstrate the chaperone roles of HSPs in the synthesis, transportation and maturity of viral proteins during viral replication in the host cells.

Objective To confirm that astrocytes from cerebral cortex of newborn rat can be the target cells of Hantaan virus (HTNV)and Seoul virus (SEOV)infection and to observe changes of astrocytes after different infection time. Methods Astrocytes were prepared from cerebral cortex of newborn rat, and then infected with HTNV and SEOV. The established virus infections were confirmed by detection of virus nucleocapsid protein (NP) and S segment RNA in astrocytes using double-label immunofluoreseence, Western-blot and reverse transcriptase-polymerase chain reaction (RT-PCR). Results The astrocytes from cerebral cortex of newborn rat were cultured successfully in vitro, which could be infected by HTNV and SEOV. The number of infected astrocytes and the virus titer in the infected astrocytes kept on increasing along with the extended infection duration. Conclusions Astrocytes from cerebral cortex of newborn rat are the target cells for HTNV and SEOV infection. Then establishment of in vitro cultured astrocytes model for Hantaviruses infection will be helpful for the study on the pathogenesis of hemorrhagic fever with renal syndrome.

Bone Neoplasms ; pathology ; Chondrosarcoma, Mesenchymal ; pathology ; Dura Mater ; pathology ; Humans ; Meninges ; pathology

Carcinoma ; pathology ; Humans ; Paranasal Sinus Neoplasms ; pathology

Objective:To study heat shock proteins(HSP) and their association with Hantavirus nucleocapsid protein(HV-NP) in Vero-E6 cells infected with Hantaan76-118.Methods: The HV-NP was identified by immunocytochemical staining after infected with Hantaan76-118.The expression of HSP was detected by Western-blot and analyzed by double specific antibody sandwich ELISA.Results: Western-blot exhibited that the expressions of HSP27,HSP70 and Grp94 in the Vero-E6 cells infected with Hantaan 76-118 were significantly higher than those in the control cells(P

Objective:To prepare polyclonal antibody against human immunoglobulin G(human IgG) and to use this antibody in the clinical diagnosis of nephrosis. Methods:New Zealand rabbit was immunized subcutaneously with human IgG.Enzyme linked immunosorbent assay(ELISA) was applied to reveal the titer of the prepared polyclonal antiserum against human IgG.Antiserum was purified with affinity chromatography,and the purified antibody was confirmed for its specificity by Western blot and immunohistochemical staining.The purified antbodies which have been indentified were fused with FITC(fluorescein isothiocyanate) and then analyzed by immunohistochemical staining.Finally the fused antibody was useed in the clinical diagnosis of nephrosis. Results: The titer of the obtained antiserum was up to(1∶128 000,) double agar diffusion test showed the titer of the antibody was 1∶32.By the method of affinity chromatography,we obtained purified antibody with the purity of 85%.ELISA,double agar diffusion and immunohistochemical staining tests showed that the specificity and titer of the antibody were not decreased sharply.The purified antibody fused with FITC also kept the specificity of the primitive antibody.When the FITC fused antibody was tried in nephrosis patients,it detected human IgG effectively. Conclusion:The polyclonal antibody can specifically recognize human IgG.This purified antibody fused with FITC can be used in the diagnosis of nephrosis.

ADP-ribosyl Cyclase 1 ; metabolism ; Antineoplastic Combined Chemotherapy Protocols ; therapeutic use ; Biomarkers, Tumor ; metabolism ; Biopsy ; Bone Neoplasms ; drug therapy ; metabolism ; pathology ; CD56 Antigen ; metabolism ; Humans ; Ilium ; Interferon Regulatory Factors ; Male ; Middle Aged ; Multiple Myeloma ; drug therapy ; metabolism ; pathology ; Syndecan-1 ; metabolism

Objective: To observe the targeting therapy of the hdsFv-RC-RNase recombinant single chain immunotoxin on the xenograft of the human hepatocellular carcinoma in nude mice and to explore its clinical potentiality. Methods: The prokaryotic expression vector TIG-hdsFv-RC-RNase was transformed into E.coli BL21(DE3)plys to largely express recombinant single chain immunotoxin hdsFv-RC-RNase against hepatocellular carcinoma induced by IPTG. The expressed product was purified via Ni-NTA affinity chromatography under native conditions and mildly refolded. The ELISA was used to analyze its immunological activity of antigen-binding capability. The xenogrft model of the human hepatocellular carcinoma in nude mice was established and the targeting therapy of hdsFv-RC-RNase was evaluated. Results:After induced by IPTG, a new protein band with M_r 41 000 was found in the supernatant of the bacteria and expressed in a soluble form. The expressed product was purified to homogeneity via Ni-NTA affinity chromatography under native conditions. The results of ELISA showed the refolded hdsFv-RC-RNase had the specific antigen-binding capability to the human hepatocellular carcinoma cell, but not to the normal hepatocyte (P