Objective:To investigate the autophagy level of CD8+T cells in rat model of chronic obstructive pulmonary disease(COPD)and correlation with its number,and to explore the role of autophagy in pathogenesis of COPD.Methods:Thirty-six 6-week-old male SD rats were divided into three groups,including normal control group,COPD group and COPD intervention group.COPD model was established by smoking,and intraperitoneal injection of 3-methyladenine(3-MA)was used for intervention.Lung function of rats was detected by small animal pulmonary function testing,and frequency of CD8+T cells,CD8+memory T cells,CD8+effector T cells in blood and lung of rats were detected by flow cytometry.CD8+T cells were sorted by immunomagnetic beads,and expressions of autophagy protein including LC3-Ⅱ/Ⅰ,Beclin-1 and p62 in CD8+T cells were detected by Western blot.The middle lobe of right lung was collected for histological observation.Results:Infiltration of inflammatory cells and the decline of lung function in COPD group were more obvious than those in normal group.Frequency of CD8+T cells,CD8+effector T cells and CD8+memory T cells in blood and lung of rats in COPD group were significantly higher than those in normal control group(all P<0.05).Level of autophagy protein,LC3-Ⅱ/Ⅰ and Beclin-1 of CD8+T cells in COPD group were significantly higher than that in normal control group,while level of p62 was lower than that in normal control group(P<0.01).CD8+T cells,CD8+effector T cells,CD8+memory T cells in COPD intervention group were significantly lower than those in COPD group(P<0.05).Correlation analysis showed that LC3-Ⅱ/Ⅰ expression level was positively correlated with frequency of CD8+T cells in blood(r=0.667,P<0.01).Expression level of Beclin-1 was positively correlated with frequency of CD8+T cells and CD8+effector T cells in blood(r=0.505,P=0.021;r=0.428,P=0.037).Expression of LC3-Ⅱ/Ⅰprotein was positively correlated with frequency of CD8+T cells and CD8+effector T cells in lung tissue(r=0.474,P=0.019;r=0.549,P=0.006).Expression of Beclin-1 was positively correlated with frequency of CD8+effector T cells in lung tissue(r=0.458,P=0.025).Conclusion:Level of CD8+T cell autophagy is correlated with its number.CD8+T cell autophagy may be involved in the chronic inflam-matory process of COPD,and 3-MA can inhibit COPD inflammation.

Objective:To investigate the autophagy level of CD8+T cells in rat model of chronic obstructive pulmonary disease(COPD)and correlation with its number,and to explore the role of autophagy in pathogenesis of COPD.Methods:Thirty-six 6-week-old male SD rats were divided into three groups,including normal control group,COPD group and COPD intervention group.COPD model was established by smoking,and intraperitoneal injection of 3-methyladenine(3-MA)was used for intervention.Lung function of rats was detected by small animal pulmonary function testing,and frequency of CD8+T cells,CD8+memory T cells,CD8+effector T cells in blood and lung of rats were detected by flow cytometry.CD8+T cells were sorted by immunomagnetic beads,and expressions of autophagy protein including LC3-Ⅱ/Ⅰ,Beclin-1 and p62 in CD8+T cells were detected by Western blot.The middle lobe of right lung was collected for histological observation.Results:Infiltration of inflammatory cells and the decline of lung function in COPD group were more obvious than those in normal group.Frequency of CD8+T cells,CD8+effector T cells and CD8+memory T cells in blood and lung of rats in COPD group were significantly higher than those in normal control group(all P<0.05).Level of autophagy protein,LC3-Ⅱ/Ⅰ and Beclin-1 of CD8+T cells in COPD group were significantly higher than that in normal control group,while level of p62 was lower than that in normal control group(P<0.01).CD8+T cells,CD8+effector T cells,CD8+memory T cells in COPD intervention group were significantly lower than those in COPD group(P<0.05).Correlation analysis showed that LC3-Ⅱ/Ⅰ expression level was positively correlated with frequency of CD8+T cells in blood(r=0.667,P<0.01).Expression level of Beclin-1 was positively correlated with frequency of CD8+T cells and CD8+effector T cells in blood(r=0.505,P=0.021;r=0.428,P=0.037).Expression of LC3-Ⅱ/Ⅰprotein was positively correlated with frequency of CD8+T cells and CD8+effector T cells in lung tissue(r=0.474,P=0.019;r=0.549,P=0.006).Expression of Beclin-1 was positively correlated with frequency of CD8+effector T cells in lung tissue(r=0.458,P=0.025).Conclusion:Level of CD8+T cell autophagy is correlated with its number.CD8+T cell autophagy may be involved in the chronic inflam-matory process of COPD,and 3-MA can inhibit COPD inflammation.

Objective To establish a new program of blood cell processing apparatus(NGL-BBS)for the preparation of washed mixed plateletsand to study the effect on biological activity of platelets compared with tradi-tional manual method.Methods Mixed concentrated platelets were separated and prepared from whole blood by white membrane method.Blood cell processing apparatus with new program set(experimental group)and manual method(control group)was used for thepreparation of washed mixed platelets.The expression rate of CD62P and CD63 in the two groups of washed mixed platelets was compared by flow-cytometry.Thrombus elastography(TEG)was used to measure and compare the MA value between the two groups.Results The expression rate of CD62P and CD63 in the experimental group was lower than that in the control group(t = 4.11,P<0.01;t = 10.78,P<0.01).TheTEG MA valueof the experimental group was higher than that of the control group(t = 6.67,P<0.01).Conclusion The present study demonstrates that the use of NGL-BBS for the preparation of washed mixed plate-lets has a lesser impact on biological activity compared to manual preparation methods.

Objective:To investigate effects of short-term cigarette smoke exposure combined with poly(I:C)stimulation on lung immune response and interferon expression in mice.Methods:BALB/c mice were randomly divided into 4 groups:control group,smoke group,poly(I:C)group and smoke combined poly(I:C)group.Total cell number and cell classification count of bronchoalveo-lar lavage fluid(BALF)were detected,and cell morphology was observed under ordinary light.Cytokines,chemokines,interferon and interferon stimulating genes expressions in lung tissues were detected by fluorescence quantitative PCR.Results:Compared with control group,total cell count,macrophage count and neutrophil count in smoke combined poly(I:C)group were significantly increased(P<0.05),and macrophage count was higher than that in poly(I:C)group.Macrophages of airway lavage fluid of mice in smoke combined with poly(I:C)group were larger in size,round or irregular in shape,and had more vacuoles in cytoplasm.Com-pared with control group,mRNA expressions of neutrophil chemokine CXCL1(P<0.05),CXCL2(P<0.01)and lymphocyte chemo-kine CCL2(P<0.01)in lung tissues of mice in smoke combined with poly(I:C)group were increased.IL-1β,IL-6,TNF-α mRNA expressions were significantly increased(P<0.01),IFN-β(P<0.01),IFN-γ(P<0.05),MX2(P<0.01)and IP-10(P<0.01)expre-ssions in lung tissues were significantly increased,and compared with poly(I:C)group,mRNA expressions of CXCL2(P<0.05),TNF-α(P<0.01)and IFN-β(P<0.05)in lung tissues of mice in smoke combined with poly(I:C)group were significantly increased.Conclusion:Cigarette smoke combined with poly(I:C)induces lung inflammation and expressions of interferon and interferon stimu-lating genes in mice.Cigarette exposure also increases poly(I:C)-induced acute lung inflammation and type Ⅰ interferon expression in mice.

Objective To explore the value of contrast-enhanced ultrasound (CEUS) in differential diagnosis of different molecular subtypes of breast cancer . Methods Sixty-two cases breast cancer patients with 62 breast lesions confirmed as breast cancer by postoperative pathology were selected . Referencing of St . Gallen standard and according to immunohistochemical markers of estrogen receptor ( ER) ,progesterone receptor(PR)andhumanepidermalgrowthfactorreceptor2(Her-2)expression,allthecasesweredivided into Luminal epithelium A or B ( Luminal A/B) subtype( 26 patients ) ,Her-2 over-expression subtype( 16 patients) and triple negative ( TN ) subtype ( 20 patients ) . CEUS and routine ultrasonography were performed for all patients before surgery ,and the contrast enhancement patterns and perfusion parameters were recorded . Whether there was a difference between the contrast enhancement patterns and perfusion parameters in different subtypes of breast cancer was analyzed . Results ① The contrast enhancement pattern showed more radioactive gathering( 76 .9% , P ＜0 .05) ,lower perfusion(69 .2% , P ＜ 0 .05) ,and lower maximum intensity ( IMAX) in the Luminal epithelium subtype than those in the Her-2 over-expression subtype and the TN subtype(all P ＜0 .05) ;② The contrast enhancement pattern showed more cardiac enhancing (93 .8% , P ＜0 .05) ,perfusion defect(75 .0% , P ＜0 .05) ,and shorter peak time (TTP) in Her-2 over-expression subtype than those in the Luminal epithelium subtype and the TN subtype(all P ＜0 .05) ;③The pattern of the TN subtype contrast enhancement showed more clear boundary( 80 .0% ,P＜0 .05) ,comparing to the other two types ,the perfusion parameters was no significant different( P ＞0 .05). Conclusions Different molecular subtypes of breast cancer have different contrast enhancement patterns and perfusion parameters ,CEUS can provide valuable imaging information for the diagnosis of breast cancer molecular typing before surgery .