ObjectiveTo establish a methodology for alveolar macrophages (AMs) phagocytosis of AlexaFluor 488 (AF488) labeled bacteria by flow cytometry.MethodsStaphylococcus aureus and Streptococcus pneumoniae were labeled with different concentrations of AF488. A flow cytometric assay was used to quantify in vivo bacterial uptake by AMs. AMs and different ratio of fluorescent-labeled bacteria were incubated at 37 ℃ for 2 hours, 4 hours, 6 hours and 8 hours, respectively. AMs were washed with DPBS and extracellular fluorescence was quenched with 1% (w/v) trypan blue. Trypan blue was aspirated and phagocytosis of fluorescent-labeled bacteria by AMs was measured using a flow cytometry. Confocal microscopy was performed to ensure that bacterial in positive AM had been internalized rather than bound to the cell surface.ResultsWhen the concentration of AF488 was more than 50 μg/mL, the labeling rates of Staphylococcus aureus and Streptococcus pneumoniae were higher than 92% (P<0.05), and has quickly reached the upper limit. With the prolongation of incubation time, the phagocytic rate of AMs increased from 20.4% at 2 hours to 76.5% at 8 hours. With the increase in the number of bacteria, the phagocytic rate of AMs increased from 7.7% by ratio of 1∶10 to 85.1% by ratio of 1∶300.ConclusionDetection of AMs phagocytosis of AF488 labeled bacteria by flow cytometry is an effective method, but the dye concentration, incubation time and the proportion of bacteria will influence the results.
Objective To observe the effects of atmospheric particulate matter (PM) exposure from two sources of motor vehicle exhaust (MVE) and biomass fuel (BMF) on the heart and lung tissues of rats. Methods Thirty rats were randomly divided into 3 groups, and they were exposed to clean air (control group), MVE (MVE group) and BMF (BMF group) for 12 weeks, respectively. The effect of long-term PM exposure on the morphological structure of heart and lung tissues was investigated by tissue and cell morphological observations, Masson staining, and immunohistochemistry. ResultsCompared with rats in the control group, the rats in the MVE group and the BMF group showed cellular degeneration, increased number of interstitial inflammatory cells, fibrous tissue proliferation, and cell apoptosis in the heart and lung tissues of the PM-exposed rats. ConclusionExposure to air pollution particulate matter for 12 weeks can significantly cause inflammatory and damage to the heart and lung tissues of rats.
ObjectiveTo detect the level of oxidative stress markers in serum, including malondialdehyde (MDA), protein carbonyls (PC), 8-hydroxy-2'-deoxyguanosine (8-OHdG) and total antioxidant capacity (TAC), in patients with stable chronic obstructive pulmonary disease (COPD), and explore the impacts of oxidant/antioxidant imbalance in pathogenesis of COPD. MethodsTwo hundred stable COPD patients (the COPD group) and 100 healthy individuals (the control group) were recruited in the study. The concentrations of MDA, PC, 8-OHdG and TAC in serum were detected. Pulmonary function test was performed and the general informations for each subjects were collected. The COPD patients were divided into a smoking subgroup and a non-smoking subgroup, or divided into a mild-moderate airflow limitation subgroup and a severe-extremely severe airflow limitation subgroup. ResultsThe levels of serum MDA, PC and 8-OHdG in the COPD group were significantly higher than those in the control group (all P < 0.01), but the level of serum TAC was significantly lower than that in the control group (P < 0.01). In the COPD patients, the levels of PC and 8-OHdG in the smoking subgroup were significantly higher than those in the non-smoking subgroup (both P < 0.05). The level of PC in the severe-extremely severe airflow limitation subgroup was significantly higher when compared with the mild-moderate airflow limitation subgroup (P < 0.01). Multiple linear regression analysis showed that the levels of PC and 8-OHdG were negatively related with FEV1% pred in the COPD patients, and the PC had greater impacts than 8-OHdG (β=-0.230, -0.219, P < 0.01). ConclusionSmoking can induce the abnormal increase of PC and 8-OHdG in serum which are negatively related with FEV1% pred in COPD patients, which suggests that oxidative stress might play an important role in pathogenesis of COPD.
目的探討經臍單孔腹腔鏡膽囊切除術（LC）的臨床應用。方法分析我院2009年1月至2010年5月期間120例因結石性膽囊炎和膽囊息肉行經臍單孔LC患者的臨床資料。結果98例患者手術成功，手術時間38～126 min，平均50.3 min。22例單孔手術失敗改成兩孔完成手術。住院時間2～4 d，平均2.5 d。全組患者無出血及漏膽并發癥發生，僅2例（1.7%）臍部戳孔處術后輕度感染，經局部換藥治療2周愈合。89例（90.8%）采用單孔法患者獲得1～15個月（平均7.3個月）隨訪，均無并發癥發生。結論單孔LC安全可行，但使用現有腹腔鏡設備操作難度較大，器械及技術尚需進一步完善。
ObjectiveTo establisht a gut microbiota mice model for chronic obstructive pulmonary disease (COPD) with fecal microbiota transplantation (FMT) and its evaluation.MethodsThe mice received FMT from healthy individuals, COPD Ⅰ-Ⅱ subjects, or COPD Ⅲ–Ⅳ subjects. After microbiota depletion, the FMT was performed by a single oral administration of 100 μL per mouse every other day, for a total of 14 times in 28 days. On the 29th day, the peripheral blood mononuclear cells were analyzed, the gut microbiota of mice before and after FMT was analyzed by 16S rRNA sequencing, and the mice model were evaluated.ResultsThe operational taxonomic units, Chao 1 and Shannon indexes of mice all decreased significantly after antibiotic treatment (P<0.001), but increased significantly after FMT from healthy individuals, COPD Ⅰ-Ⅱ subjects, or COPD Ⅲ–Ⅳ subjects (P<0.05 or P<0.01). The abundance of Firmicutes, Proteobacteria and Actinobacteria in the guts of the mice in the healthy human FMT group, COPD Ⅰ-Ⅱ FMT group and COPD Ⅲ-Ⅳ FMT group were significantly different from those of the control group who only received phosphate buffer saline instead of FMT (P<0.05 or P<0.01). The auxiliary T lymphocytes and cytotoxic T lymphocytes were higher, but B lymphocytes decreased in the peripheral blood of the mice in the COPD Ⅰ-Ⅱ FMT group and COPD Ⅲ-Ⅳ FMT group (P<0.05 or P<0.01).ConclusionFMT can successfully establish a COPD gut microbiota research model.