Chloro-nitrobenzene compounds air sampling tube preparation and determination method / 中国职业医学

@#Objective - - To prepare the GDH 5 air sampling tube for simultaneous collection of eight kinds of chloro nitrobenzene （ ） ， compounds CNBs in the air of workplace and establish a matching determination method using gas chromatography. Methods - - ， Eight kinds of CNBs in vapor and aerosol state were collected by self developed GDH 5 air sampling tube desorbed ， ， ， by toluene separated by polysiloxane gas chromatography column detected by microcell electron capture detector and Results - （ - quantified by external standard method. It was determined that the air sampling tube was assembled by XAD 2 ion ） - ， exchange resin and glass fiber filter membrane. The linear range of CNBs was 0.80 240.00 mg/L and the linear correlation - - coefficients were greater than 0.999 9. The detection limit was 7.87 13.03 μg/L. The minimum detectable concentration was 0.60 3， - 3（ ） 1.33 μg/m and the minimum quantitative concentration was 2.00 4.22 μg/m sample 45.00 L . The average desorption - - （RSD） - ， - RSD efficiency was 101.2% 110.0%. The within run relative standard deviation was 0.8% 4.1% and the between run - Conclusion - was 0.3% 5.8%. The samples could be stored for more than 30 days at room temperature. GDH 5 air sampling tube and its associated determination method can be used for the collection and determination of eight kinds of CNBs in workplace air.

Inactivated Sendai virus suppresses murine melanoma growth by inducing host immune responses and down-regulating β-catenin expression / 生物医学与环境科学（英文）

<p><b>OBJECTIVE</b>This paper aims to investigate the anti-tumor mechanism of inactivated Sendai virus (Hemagglutinating virus of Japan envelope, HVJ-E) for murine melanoma (B16F10).</p><p><b>METHODS</b>The murine dendritic cells (DCs) were treated with HVJ-E, and then the cytokines secreted from DCs and costimulation-related molecules on DCs were measured. Meanwhile, the expression of β-catenin in HVJ-E treated murine melanoma cells was detected. In addition, HVJ-E was intratumorally injected into the melanoma on C57BL/6 mice, and the immune cells, CTL response and tumor volume were analyzed.</p><p><b>RESULTS</b>HVJ-E injected into B16F10 melanoma obviously inhibited the growth of the tumor and prolonged the survival time of the tumor-bearing mice. Profiles of cytokines secreted by dendritic cells (DCs) after HVJ-E stimulation showed that the number of cytokines released was significantly higher than that elicited by PBS (1P<0.05). The co-stimulation-related molecules on DCs were comparable to those stimulated by LPS. Immunohistochemical examinations demonstrated the repression of β-catenin in B16F10 melanoma cells after HVJ-E treatment. Meanwhile, real-time reverse transcription PCR revealed that HVJ-E induced a remarkable infiltration of CD11c positive cells, chemokine ligand 10 (CXCL10) molecules, interleukin-2 (IL-2) molecule, CD4(+) and CD8(+) T cells into HVJ-E injected tumors. Furthermore, the mRNA expression level of β-catenin in the HVJ-E injected tumors was also down-regulated. In addition, B16F10-specific CTLs were induced significantly after HVJ-E was injected into the tumor-bearing mice.</p><p><b>CONCLUSION</b>This is the first report to show the effective inhibition of melanoma tumors by HVJ-E alone and the mechanism through which it induces antitumor immune responses and regulates important signal pathways for melanoma invasion. Therefore, HVJ-E shows its prospect as a novel therapeutic for melanoma therapy.</p>

Gefitinib inhibits α-smooth muscle actin expression in mice with bleomycin-induced lung fibrosis / 南方医科大学学报

<p><b>OBJECTIVE</b>To evaluate the effect of epidermal growth factor receptor tyrosine kinase inhibitor, gefitinib, on the expression of α-smooth muscle actin (α-SMA) in mice with lung fibrosis induced by bleomycin.</p><p><b>METHODS</b>Thirty male BALB/c mice were randomly divided into control, bleomycin, and bleomycin plus gefitinib groups. The mice in the control group were subjected to intratracheal administration of normal saline, those in bleomycin group received bleomycin (3 mg/kg) intratracheally, and those in bleomycin plus gefitinib group received oral gefitinib (20 mg/kg administering) plus intratracheal bleomycin administration. All the mice were sacrificed 14 days after the treatments, and the left lung was examined pathologically with HE staining and Masson staining and also immunohistochemically for assay of the total EGFR, phosphorylated EGFR and α-SMA. The right lungs were sampled for RT-PCR to detect the mRNA levels of α-SMA.</p><p><b>RESULTS</b>Gefitinib administration lessened lung fibrosis induced by bleomycin and significantly reduced lung collagen accumulation. The phosphorylation of EGFR in the pulmonary mesenchymal cells and epithelial cells and the expression levels of α-SMA mRNA and protein were inhibited by gefitinib treatment in mice with intratracheal administration of bleomycin (P<0.05).</p><p><b>CONCLUSION</b>Gefitinib offers protection against lung fibrosis induced by bleomycin in mice probably by inhibiting the downstream signals of EGFR and by downregulating the expression of α-SMA.</p>

Construction and identification of recombinant avian adeno-associated virus expressing GFP reporter gene / 病毒学报

To generate recombinant avian adeno-associated virus (rAAAV) for gene transfer studies in avian cells, the recombinant plasmid containing the whole genome of AAAV was digested with restriction enzymes to remove the Rep and Cap genes, resulting in AAAV transfer vector pAITR. GFP-expressing cassette was amplified by PCR and inserted into the AAAV transfer vector. The Rep-Cap gene of AAAV amplified by high fidelity PCR was subcloned into eukaryotic expression vector pcDNA3, resulting in an AAAV helper vector pcDNA-ARC. The Rep and Cap genes amplified by high fidelity PCR were subcloned separately into the co-expression vector pVITRO2-mcs, resulting in another AAAV helper vector pVITRO2-ARC. Using calcium phosphate precipitation method, rAAAV-GFP was generated by co-transfecting AAV-293 cells with a cocktail of pAITR-GFP, pcDNA-ARC or pVITRO2-ARC, and adenovirus helper vector pHelper. The three structural proteins VP1, VP2 and VP3 of correct molecular masses were detected by SDS-PAGE and the GFP reporter gene was detected by PCR in purified rAAAV-GFP virions. Chicken embryonic fibroblast (CEF) cells and CEL cell line were transduced with the recombinant virus, the GFP-positive cells were easily observed under fluorescent microscope, expression of which lasted for at least two weeks. These data demonstrate that an efficient helper virus-free packaging system has been established for generating recombinant AAAV particles for gene transfer studies in avian cells and for development of recombinant vaccines against avian diseases.