Pulmonary pathology in fatal human influenza A (H1N1) infection / 中华病理学杂志

<p><b>OBJECTIVE</b>To study the pulmonary pathology in patients died of fatal human influenza A(H1N1) infection.</p><p><b>METHODS</b>Eight cases of fatal human influenza A (H1N1) infection, including 2 autopsy cases and 6 paramortem needle puncture biopsies, were enrolled into the study. Histologic examination, immunohistochemitry, flow cytometry and Western blotting were carried out.</p><p><b>RESULTS</b>The major pathologic changes included necrotizing bronchiolitis with surrounding inflammation, diffuse alveolar damage and pulmonary hemorrhage. Influenza viral antigen expression was detected in the lung tissue by Western blotting. Immunohistochemical study demonstrated the presence of nuclear protein and hemagglutinin virus antigens in parts of trachea, bronchial epithelium and glands, alveolar epithelium, macrophages and endothelium. Flow cytometry showed that the apoptotic rate of type II pneumocytes (32.15%, 78.15%) was significantly higher than that of the controls (1.93%, 3.77%).</p><p><b>CONCLUSION</b>Necrotizing bronchiolitis, diffuse alveolar damage and pulmonary hemorrhage followed by pulmonary fibrosis in late stage are the major pathologic changes in fatal human influenza A (H1N1) infection.</p>

The effect of 5-fluorouracil on enriching cancer stem cells of hepatoma cell line BEL-7402 / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To investigate the effect of 5-FU (5-fluorouracil) on enriching cancer stem cells of HCC cell line BEL-7402 and the biological characteristics of enriched cells.</p><p><b>METHODS</b>The enriching concentration of 5-FU was determined by CCK-8 (cell counting kit-8). Flow Cytometry was used to determine the changes in cell cycle and positive expression ratio of surface marker CD56, CD54, EpCAM and CD133. The self-renewal and differentiation of positive cells were tested by colony formation assay, and were compared with the control group.</p><p><b>RESULTS</b>Enriching concentration of 5-FU was determined as 10 μg/ml with 48 h incubation. After enrichment, G0/G1 phase cells increased from 57.50 %+/-0.98% to 68.70%+/-3.41% (P<0.05). Whereas S phase cells decreased from 40.26%+/-4.12% to 31.80%+/-4.15% (P<0.01); G2/M phase cells disappeared in experimental group, and was 5.80%+/-1.87% in control group (P<0.01). The proportion of the cell cycle changed with significant statistical differences. Meanwhile, positive rate of cell surface makers CD56, CD54, EpCAM and CD133 increased from 0.57%+/-0.12%, 8.10%+/-6.79%, 0.3%+/-0.01% and 3.20%+/-0.99% to 4.13%+/-0.06%, 50.08%+/-1.69%, 0.55%+/-0.07% and 10.51%+/-1.13%, respectively. The difference was significant (P<0.05). The colony forming ratio of CD56, CD54, EpCAM and CD133 negative cells and positive cells were 2.11%+/-0.21%, 3.32%+/-0.31%; 0.86%+/-0.101%, 2.40%+/-0.52 %; 7.19%+/-0.56%, 7.73%+/-0.71%; 2.70%+/-0.26%, 5.75%+/-0.81%, respectively, and significant differences were found between (P<0.05).</p><p><b>CONCLUSION</b>5-fluorouracil enriched the cancer stem cell population in HCC cell line BEL-7402. CD56 and CD54 can be used as important surface markers in research of liver cancer stem cells.</p>

Experimental research of discordant tracheal xenotransplantation / 中华整形外科杂志

<p><b>OBJECTIVE</b>To investigate the immunological rejection mechanism of tracheal xenotransplantation and xenografts as potential sources of trachea.</p><p><b>METHODS</b>On SD rat model, a xenotransplanted tracheal from the guinea pig was established by wrapping it in the cervical muscles in situ. It was divided into cryopreserved group and uncryopreserved group. Under the examinations with histochemistry, immunofluorescence (IFL) and flow cytometry (FCM) techniques, the pathomorphological characteristics of the tracheal xenografts and the immunological rejection mechanism were evaluated.</p><p><b>RESULTS</b>The tracheal allotransplantation with cryopreserved grafts wrapped by neck muscles was survived for a longer period. Histological examination revealed normal appearance of the allografts. The tracheal grafts patency was above 80%. However, cryopreserved tracheal xenografts of the guinea pig-to-rat maintained vitality for 14 days in maximum and 13.2 days on average, while the fresh tracheal xenografts only for 9 days in maximum, and 8 days on average. Acute rejection occurred in the tracheal xenotransplantation. A marked mononuclear-macrophage cellular infiltration mixed with eosinophils and lymphocyte was seen in the xenografts. Antibody (IgM, IgG) and complement (C3) deposition were also obviously detected by IFL in the xenografts. CD4 T+ cells and CD8+ T cells increased significantly in the vascular circulation. In all of the xenografts, complete loss of tracheal epithelium was associated with cartilage necrosis. The grafts patency was below 50%. This performance deteriorated with extended time periods. The fresh xenografts performed significantly worse than the cryopreserved xenografts.</p><p><b>CONCLUSIONS</b>Acute rejection, caused by humoral immune reaction mainly integrated with cellular immunity, is the most notable characteristics in the guinea pig-to-rat tracheal xenotransplantation in situ. Cryopreservation can potentially reduce the antigenicity. The low antigenicity may inhibit the immunologic reaction relatively, so that prolonged survival of discordant cryopreserved tracheal xenografts could be achieved.</p>