Objective To observe the rat tracheal stem cells in situ.Methods Tracheal rings of Wistar rats treated with 5-FU to make an injure model in vitro then observing the process of regeneration in sequence time from 3*!hours to 48*!hours. Use light microscope to detect the development of tracheal epithelium,assay the level of proliferating cell nuclear antigen (PCNA) immunohistochemistry and perform Hoechst33342 staining to search the negative staining cells. Results Twelve hours after 5-FU it could be seen that tracheal mucosa exfoliated with nuke-nucleus like cells nailed just above the basement membrane,6*!hours after removing 5-FU we could see that the tracheal rings were covered with squamous epithelium. It could be seen that PCNA immunohistochemistry negative staining cells located bordering on the positive one. Meanwhile under fluorescent contrast microscope negative cells with no Hoechst dye combinations were among most of the positive cells. 12*!hours after removing 5-FU mucous granular cells and cilia cells could be seen. After 48*!hours the whole tracheal rings were covered with pseudostratified columnar ciliated epithelium.Conclusion 5-FU is a cell-cycle specific antimetabolite cytotoxicant.It can make the cycling cells degeneration and necrosis but has little effect on the G 0 cells. Few G 0 cells which look like nuke nucleus cells randomly on the basement have the ability to efflux the dye Hoechst33342.The tracheal rings are regenerated through these cells differentiation and proliferation.

Objective To analyze the condition of Rhodamine 123 staining of human bronchial epithelium in vitro during the wound-repair process induced by fluorouracil(5-FU). Methods The bronchial epithelial cells in normal and 5-FU injurious group were obtained by enzymatic digestion and analyzed by flow cytometry.1.PI staining to identify the percentage of cells in different cell phase;2.Rhodamine 123 staining in live cells was to contrast the percentage of negative cells in two groups. Results 1.Analysis of cell cycle:apoptotic cells in injurious group increased,cells in S+G-2/M phase almost disappeared,majority of the remain live cells of injurious group were in G-0/G-1 phase;with the recover of bronchial epithelium,cells in S+G-2/M phase increased;2.The percentage of Rhodamine 123 negative staining live cells in injurious group was higher than that of normal group,and with the recover of bronchial epithelium,this percentage lowered to nearly normal.Conclusion 5-FU can make the cycling cells apoptosis and reserve quiescent phase cells.Among them there were bronchial stem cells which had the capacity to efflux Rhodamine 123.Just the proliferation and differentiation of these stem cells regenerated bronchial epithelium.Rhodamine 123 staining to sort the remain cells after 5-FU treatment by FACS can be used to enrich and purify bronchial stem cells.

10.3969/j.issn.2095-4344.2013.23.023

Objective: Our aim was to discuss the pathological characteristics and immunophenotype of non-Hodgkin lymphoma (NHL) in Shenyang. Methods: Histopathological observation was performed with immunohistochemical methods (SP method). We used 10 antibodies as B,T markers to analyze 76 cases of NHL according to the new WHO classification. Results: The B-NHL was more prevalent than the T-NHL. In B-NHL, the Diffuse Large B cell Lymphoma (DLBL) was the most common, next was the mucosa-associated lymphoid tissue lymphoma (+/-monocytoid B-cells) and lymphoplasmacytic lymphoma (LPL). In T-NHL, the peripheral T cell lymphoma unspecificly accounted most. The positivity rates of CD79a in B-NHL were 100% and its cross-reactivity was 0. The straining rates of polyclonal CD3 with T cell were 88% and CD3 only reacted with 5% of B-NHL. Conclusion: The NHL in Shenyang was marked by the most common DLBL and peripheral T cell lymphoma, unspecificly. The rates of monocytoid lymphoma and LPL were higher. The CD79a and CD3 were antibodies of B-cell and T-cell markers with high sensitivity and specificity, respectively.

Objective To explore the dynamic changes of tracheal stem cells during tracheal regeneration after injury induced by fluorouracil(5-FU) in rats. Methods Extracorporeal tracheal injury(Wistar rats) was induced by 5-FU.ABCG2 expression in tracheal epithelium during the process of regeneration was analyzed by indirect immunofluorescence and Western blotting. Results 1.After treatment with 5-FU for 12 hours,the tracheal epithelium shed and there were ABCG2 positive cells among residual cells in G-0.Three hours after the removal of 5-FU,the tracheal rings were covered with flattened epithelial cells. ABCG2 positive cells increased slightly.Six-nine hours after the removal of 5FU,the epithelial cells changed into cuboidal,correspondingly,the ABCG2 positive cells increased obviously;At 24 hours after the removal of 5-FU,most of the epithelial cells were cuboidal and merged into pieces,but the ABCG2 positive cells decreased obviously;Until 48 hours after the removal of 5-FU,only a few positive cells could be seen with the pseudostratified mucociliary epithelium restored similar to its original mode.There were no detectable ABCG2 positive cells in normal tracheal epithelium.2.Western blotting analysis showed that there were different ABCG2 levels at different times after the removal of 5-FU which in accordance with the change of immunofluorescene.ABCG2 was minimally detected after treatment with 5-FU for 12 hours,reaching a maximal level at 6 hours after the removal of 5-FU,and then decreased over time.Until 48 hours after the removal of 5-FU,very low ABCG2 level was detected.Conclusion The expression of ABCG2 is correlated to the number of tracheal stem cells,suggesting that ABCG2 may serve as a marker for isolating stem cells from tracheal epithelium.

Objective\ To detect thrombomodulin protein in 18 weeks human fetal lung. Methods\ SP method was used in the study. Results\ It showed that, in fetal lung tissues, thrombomodulin expressed in the endothelial cells of capillaries surrounding the primary alveoli, but it was absent in cuboid cells and ciliated\|columnary epithelium cells and cartilage. Conclusion\ Our results suggest that thrombomodulin does not exist in middle and late stage's human fetal lung.\;

Animals ; Aquaporin 3 ; analysis ; Cell Separation ; Fluorouracil ; pharmacology ; Humans ; Lung ; cytology ; Membrane Proteins ; analysis ; Octamer Transcription Factor-3 ; genetics ; Stem Cells ; cytology ; Trachea ; cytology

Animals ; Humans ; Lung Neoplasms ; metabolism ; pathology ; Neoplastic Stem Cells ; pathology

Objective To evaluate the value of perfusion imaging mismatch and low perfusion ratio(HIR)based on CT perfusion imaging in predicting acute intracranial large vessel occlusion(LVO)associated with intracranial atherosclerotic stenosis(ICAS).Methods A total of 82 pa-tients with acute intracranial LVO who underwent emergency thrombectomy in our hospital from February 2019 to December 2020 were enrolled in this study.According to the etiology,they were divided into ICAS-related LVO group(ICAS-LVO,65 cases)and cardiogenic embolism group(17 cases).ROC curve was plotted to analyze the predictive value of CT perfusion imaging parame-ters.Results Compared with the cardiogenic embolism group,the ICAS-LVO group had signifi-cantly larger male ratio,higher BMI and TG level,more severe progression of disease,longer time from onset to surgery,larger proportion of ischemic penumbra and higher mismatch ratio,and ob-viously less ratio of atrial fibrillation,lower BNP and HDL levels,smaller infarct volume,and lower HIR(P＜0.05,P＜0.01).ROC curve analysis showed that HIR and mismatch ratio had good predictive value for the etiology of ICAS-LVO.The optimal cut-off value of HIR was 0.26,with an AUC value of 0.74,a specificity of 0.88,and a sensitivity of 0.54.The optimal cutoff for the mismatch ratio was 3.84,with an AUC value of 0.84,a specificity of 0.75,and a sensitivity of 0.90.Generalized linear model revealed that HIR and cerebral blood volume index had no signifi-cant difference in prognostic performance(P=0.175).Conclusion HIR and mismatch ratio are helpful to identify the pathogenesis earlier and formulate surgical strategies more accurately,thereby reducing iatrogenic injury to a greater extent,increasing the effective reperfusion rate,re-ducing the disability and mortality,and improving the prognosis of clinical outcomes.