Dendritic cells (DCs) have an important role in initiating and maintaining the immune inflammatory response in allergic asthma, and CC chemokine receptor 7 (CCR7) is directly involved in the pathogenesis of DC- and T cell-mediated allergic asthma. The present study aimed to investigate the effects of CCR7 on DC-mediated immune tolerance in allergic asthma. In the present study, bone marrow-derived DCs were transfected with an adenovirus encoding the rat CCR7 gene or a short hairpin RNA targeting CCR7 (sh-CCR7). Rats injected with DCs overexpressing CCR7 or presenting CCR7 knockdown were examined. After the rats were injected with DCs via the tail vein, bronchoalveolar lavage fluid was collected to assess its cellular composition. The protein expression levels of CCR7 in DCs were determined using immunohistochemistry and western blot analysis. The protein expression levels of interferon-gamma (IFN-gamma), interleukin-4 (IL-4), IL-10, IL-12, transforming growth factor-beta (TGF-beta) and immunoglobulin E (IgE) were determined by ELISA. Compared with the control group, the protein expression level of CCR7 was significantly higher in the CCR7 overexpression group and significantly lower in sh-CCR7 group. Similarly, the number of DCs was higher in the CCR7 overexpression group and lower in the sh-CCR7 group. The protein expression levels of IL-10 and TGF-beta were significantly lower in the CCR7 overexpression group and higher in the sh-CCR7 group. In addition, the expression levels of IL-4, IL-12, IFN-gamma and IgE were higher in the CCR7 overexpression group and lower in the sh-CCR7 group. The present results suggested that the role of cytokines and IgE in immune inflammation and immune tolerance in allergic asthma may be associated with the expression level of CCR7 in DCs, suggesting that CCR7 may serve a role in DC-mediated immune tolerance in allergic asthma.

肺血栓栓塞症(pulmonary thromboembolism,PTE)是以各种栓子阻塞肺动脉或其分支为其发病原因的一组疾病或临床综合征的总称. 随年龄增长、肺栓塞发病率及死亡率呈上升趋势. 老年肺栓塞患者常合并多种基础心肺疾病,致使临床表现不典型、诊断困难,误诊、漏诊率很高.

Background Cigarette smoking aggravates the symptoms of asthma, leading to the rapid decline of lung function. Dendritic cells (DCs) and lymphocytes are considered initiating and promoting factors for the airway inflammation reactions of asthma. In addition, activation of CC chemokine receptor 7 (CCR7) by chemokine (C-C motif) ligand (CCL) 19 and 21 promotes DCs and T cells migration to lymphoid tissues during inflammation. We aimed to examine how cigarette smoke affects the expression of CCR7 in the lungs of asthmatic rats and explore the signaling mechanism linking CCR7 expression to exacerbation of symptoms. Methods Forty Wistar rats were randomized to four groups: control, asthma, smoke exposure, and asthma with smoke exposure groups. A rat asthma model was established by intraperitoneal ovalbumin injection. CCR7 expression was examined with immunohistochemistry and western blotting. The number of airway DCs was determined by OX62 immunohistochemistry. Interferon (INF)-γ, interleukin (IL)-4, CCL19, and CCL21 expression levels in blood and bronchioalveolar lavage fluid (BALF) were determined by enzyme-linked immunosorbent assays (ELISAs). Results Tissue CCR7 expression, peripheral blood and BALF CCL19 and CCL21 concentrations, and the number of airway DCs were significantly higher in the asthma with smoke exposure group than the asthma group (P<0.01). In addition, INF-γ expression was decreased and IL-4 increased in the asthma and asthma with smoke exposure groups compared with the control group (P<0.01), and in the asthma with smoke exposure group compared with the asthma group (P<0.01). Expression of CCR7 correlated negatively with INF-γ expression in peripheral blood and BALF (P<0.01), and positively with the airway DCs and IL-4 expression in the peripheral blood and BALF (P<0.01). Conclusions Cigarette smoking may aggravate asthma symptoms by attenuating immunity, possibly through CCR7-mediated DCs aggregation in lung tissue.

Objective To study the effect of cigarette smoke exposure on the expression of CC chemokine receptor 7 (CCR7) and its ligands of peripheral blood mononuclear cells in asthmatic patients,and to explore correlation between CCR7 and clinical targets.Methods The outpatients with asthma (asthma group,n =15) and smoking outpatients with asthma (smoking & asthma group,n =15) were enrolled in the study.Meanwhile,healthy controls (control group,n =15) and healthy smokers (smoking group,n =15)were matched.All of them required to blood test and assess asthma control test (ACT) score,blood gas analysis and lung function.CCR7 of peripheral blood mononuclear cells were measured by flow cytometry,CC chemokine ligand 21 (CCL21) and CCL19 in peripheral blood were measured by enzyme-linked immunosorbent assay.Results ① Compared with the control group,the expression of CCR7 protein increased remarkably in the asthma group and the smoking exposure combined asthma group (all P ＜ 0.01).Compared with the asthma group,the expression of CCR7 protein increased remarkably in the smoking combined asthma group (all P ＜0.01).②Compared with the control group,CCL21 and CCL19 increased remarkably in the smoking combined asthma group (all P ＜ 0.01).Compared with the asthma group,CCL21 and CCL19 increased remarkably in the smoking combined asthma group (all P ＜0.01).③Compared with the control group,forced expiratory volume in 1 second (FEV1) and peak expiratory flow (PEF) reduced remarkably in the asthma group and the smoking combined asthma group (all P ＜0.01).Compared with the asthma group,ACT score,FEV1 and PEF increased remarkably in the smoking combined asthma group (all P ＜0.01).④The expression of CCR7 protein was negatively correlated with FEV1 and PEF (all P ＜0.01).Conclusions Smoking can increase the expression of CCR7 and its ligand CCL19 and CCL21,which may be result in the aggravation of symptoms and the decline of lung function in patient with asthma.

目的 探讨CC趋化因子受体(CC chemokine receptor,CCR)7在烟草烟雾暴露加重哮喘肺部炎症中的作用.方法 40只Wistar大鼠随机分为四组:对照组(C组)、哮喘组(A组)、烟雾暴露组(S组)、烟雾暴露+哮喘组(AS组),参照文献建立各组大鼠模型.收集外周血和支气管肺泡灌洗液(Bronchoalveolar lav-age fluid,BALF),BALF中白细胞计数及分类采用HE染色,血浆和BALF中IL-12蛋白的检测采用酶联免疫吸附试验法(ELISA),肺部CCR7蛋白的表达采用免疫组织化学法检测.结果 ①AS组、A组大鼠肺部CCR7表达较C组增加,AS组大鼠肺部CCR7表达较A组增加,差异均有统计学意义(均P<0.01).②AS组、A组大鼠血浆、BALF中IL-12表达较C组降低(均P<0.01);AS组大鼠血浆、BALF中IL-12表达较A组降低(均P<0.05).③A组和AS组大鼠BALF白细胞总数、中性粒细胞、淋巴细胞比例较C组增加,差异均具有统计学意义(均P<0.01);AS组大鼠BALF嗜酸粒细胞较A组减少,白细胞总数、中性粒细胞比例较A组增加,差异均具有统计学意义(均P<0.01)④CCR7的表达与血、BALF中IL-12表达呈负相关(分别r=-0.677,r=-0.692,均P<0.01);与BALF中白细胞数量、中性粒细胞数量呈正相关(分别r=0.790,r=0.825,均P<0.01).结论 烟草烟雾暴露可增加哮喘大鼠肺部CCR7的表达,同时影响BALF细胞计数及分类,导致IL-12表达降低,提示CCR7可能在烟雾暴露加重哮喘肺部炎症的机制中发挥一定作用.

Objective: To investigate the effect of cigarette smoke exposure on the expression of CC Chemokine receptor 7 (CCR7) and levels of Th1/Th2 cytokines in asthmatic rats. Methods: Forty Wistar rats were randomly divided into four groups: control group, asthma group, smoke exposure group, asthma-smoke exposure group. The asthma group were sensitized with ovalbumin (OVA) and Aluminum hydroxide at day 1, 8 and challenged with OVA at day 15 by atomization for 8 weeks.While control group was sensitized and challenged with normal saline instead of OVA.The smoke exposure group was sensitized and challenged with normal saline instead of OVA followed passive smoking for 8 weeks. The asthma-smoke exposure group was challenged with OVA followed passive smoking. The pathological changes of different groups were observed by HE-staining. CCR7 was semiquantitatively analyzed in lungs by immunohistochemistry.The concentration of CC chemokine ligand (CCL)19, CCL21, interferon (IFN)-γ and interleukin (IL)-4 in peripheral blood and CCL19 and CCL21 in bronchoalveolar lavage fluid (BALF) were measured by enzyme-linked immunosorbent (ELESA) assay. Results: In asthma group, smoke exposure group and asthma-smoke exposure group, the various degrees of inflammatory reaction appeared in lung tissue and the asthma-smoke exposure group was with the most significant reaction. In the lung tissues of the rats from asthma group, smoke exposure group and asthma-smoke exposure group, the average optical density (AOD) of CCR7 were significantly higher than those in control group (0.350±0.023, 0.252±0.022, 0.400±0.029 vs 0.180±0.020, all P<0.01). The AOD of CCR7 of asthma-smoke exposure group was much higher than both that in asthma group and in smoke exposure group (both P<0.01). In asthma group, smoke exposure group and asthma-smoke exposure group, the concentrations of both CCL19 and CCL21 in peripheral blood and BALF were significantly higher than that in control group (all P<0.01). The concentrations of both CCL19 and CCL21 in peripheral blood and BALF of asthma-smoke exposure group were significantly higher than the results in asthma group and in smoke exposure group (all P<0.01). The concentrations of IFN-γ in peripheral blood of asthma group and asthma-smoke exposure group were lower than those in control group [(33±3), (17±3) vs (70±4) pg/ml], but asthma-smoke exposure group was much lower than the results in asthma group (all P<0.01). The concentration of IFN-γ in peripheral blood of smoke exposure group[(100±5)pg/ml]was higher than that in control group and asthma-smoke exposure group (both P<0.01). In asthma group, smoke exposure group, asthma-smoke exposure group, the concentrations of IL-4 in peripheral blood were significantly higher than those in control group [(54±4), (42±4), (76±4) vs (30±4) pg/ml, all P<0.01]. The concentrations of IL-4 in peripheral blood of asthma-smoke exposure group was significantly higher than those in asthma group and in smoke exposure group (both P<0.01). Conclusion: Cigarette smoke could enhance the expression of CCR7 and its ligand, and it can also result in exacerbations of asthma by reducing the expression level of IFN-γ (the representative of Th1 cytokine) and increasing the expression level of IL-4 (the representative of Th2 cytokine).

Hypoxic pulmonary hypertension (HPH) may contribute to vascular remodeling, and pulmonary artery smooth muscle cell (PASMC) proliferation has an important role in this process. However, no relevant information concerning the role and mechanism of protein kinase C (PKC)α in hypoxia-induced rat PASMC proliferation has been elucidated. The present study aimed to further investigate this by comparison of rat PASMC proliferation among normoxia for 72 h (21% O2), hypoxia for 72 h (3% O2), hypoxia + promoter 12-myristate 13-acetate control, hypoxia + safingol control, hypoxia + PD98059 control and hypoxia + U0126 control groups. The present study demonstrated that protein expression levels of PKCα in rat PASMCs were elevated. In conclusion, through activating the extracellular signal-regulated 1/2 signaling pathway, PKCα is involved in and initiates PASMC proliferation, thus bringing about pulmonary artery hypertension. These results add to the understanding of the mechanism PKCα in PH formation and lays a theoretical basis for prevention as well as treatment of HPH.

In chronic hypoxia, pulmonary hypertension (PH) induces right ventricular hypertrophy (RVH). Evidence indicates that protein kinase C (PKC) serves a crucial role in hypoxia-induced RVH. The present study investigated PKC isoform-specific expression and its involvement in RVH. Rats were exposed to normobaric hypoxia for a number of days to induce PH. PKC isoform-specific membrane translocation and protein expression in the myocardium were evaluated by western blotting and immunostaining. A total of six isoforms of conventional PKC (cPKC; alpha, beta I and beta II) and of novel PKC (nPKC; delta, epsilon and eta), were detected in the rat myocardium. Hypoxic exposure (1-21 days) induced PH with RVH and vascular remodeling. nPKCd membrane translocation at 3-7 days and cPKC beta I expression at 1-21 days in the RV following hypoxic exposure were significantly decreased as compared with the normoxia control group. Membrane translocation of cPKC beta II at 14-21 days and of nPKC eta at 7-21 days in the left ventricle following hypoxic exposure was significantly increased when compared with the control. The results of the present study suggested that the alterations in membrane translocation, and nPKC delta and cPKC beta I expression, are associated with RVH following PH, and the upregulation of cPKC beta II membrane translocation is involved in left-sided heart failure.

Objective To study the effect of cigarette smoke exposure on the expression of histone deacetylase (HDAC)2,interferon (IFN)-γ and interleukin(IL)-4,and to explore immunological mechanism of Th1/Th2 imbalance about cigarette smoke exposure aggravating asthma.Methods Fifty 50 healthy male Wistar rats were divided randomly into control group,asthma group,cigarette smoke exposure combined asthma group.Asthma group was treated by budesonide,cigarette smoke exposure combined asthma group treated by budesonide.We established the rats model of every group.The expression of HDAC2 protein was determined by immunohistochemistry and IFN-γ、IL-4 in peripheral blood and bronchoalveolar lavage fluid (BALF) were measured by enzyme-linked immunosorbent assay (ELISA).Results Compared with the control group,the expression of HDAC2 protein increased remarkably in the asthma group and the cigarette smoke exposure combined asthma group(P ＜ 0.01,respectively).Compared with the asthma group,the expression of HDAC2 protein increased remarkably in the cigarette smoke exposure combined asthma group(P ＜0.01).Compared with the asthma group treated by budesonide,the expression of HDAC2 protein increased remarkably in the cigarette smoke exposure combined asthma group treated by budesonide(P ＜ 0.01).Compared with the control group,IFN-γ of peripheral blood and BALF decreased and IL-4 increased remarkably in the asthma group and the cigarette smoke exposure combined asthma group(P ＜ 0.01).Compared with the asthma group,IFN-γ of peripheral blood and BALF decreased and IL-4 increased remarkably in the cigarette smoke exposure combined asthma group (P ＜ 0.01).Compared with the asthma group treated by budesonide,IFN-γ of peripheral blood and BALF decreased and IL-4 increased remarkably in the cigarette smoke exposure combined asthma group treated by budesonide (P ＜ 0.01).The expression of HDAC2 protein was positively correlated with IFN-γ of peripheral blood and BALF and was negatively correlated with IL-4 of peripheral blood and BALF.Conclusion Cigarette smoke exposure can down-regulate the expressi,on of HDAC2 protein,IFN-γ,and up-regulate the expression of IL-4,which may play an important role in aggravating airway inflammation and Th1/Th2 balance in asthmatic rats.

Objective To determine the effect of nuclear factor (NF)-κB signaling pathway in the interleukin (IL)-12 expression of dendritic cells (DCs) in rats with nicotine-induced bronchial asthma, and to investigate the pathological mechanism of asthma exacerbated by smoking. Methods The rat models of bronchial asthma were es-tablished, and the bone marrow-derived DCs were cultured in vitro. Nicotine intervention was given, under different experimental conditions. The concentration of IL-12 protein in the supernatant was measured by ELISA, and the opti-mum concentration and time of nicotine effect were determined. Furthermore, DCs were intervened with NF-κB in-hibitor, pyrrolidine dithiocarbamate (PDTC). The expression of NF-κB was determined by immunocytochemistry. The levels of IL-12 protein in the supernatant of each group were determined by ELISA, and the mixed lymphocyte reac-tion was performed. The proliferation activity of lymphocytes was determined by MTT assay. Results ①Compared with the control group, the expression of IL-12 protein in DCs increased in the 200 μg/ml nicotine group and de-creased in the 400 μg/ml nicotine group, and the difference was statistically significant (all P<0.01). ②After the DCs were incubated with 200μg/ml nicotine for 0, 4, 8, 12, 24 and 72h, the expression of IL-12 protein reached a peak at 12 h and decreased at 72 h, respectively, and the differences were statistically significant (all P<0.01 ). ③Compared with the control group, the DCs IL-12 protein expression increased in the nicotine group and decreased in the PDTC group, and the difference was statistically significant (all P<0.01). Compared with the nicotine group, the DCs IL-12 protein expression decreased in the nicotine+PDTC group, and the difference was statistically significant ( P<0.05). ④Compared with the control group, the proliferation of allogeneic lymphocytes stimulated by DCs increased in the nico-tine group, and decreased in the PDTC group, and the difference was statistically significant (all P<0.01). Compared with the nicotine group, the proliferation of allogeneic lymphocytes stimulated by DCs decreased in the nicotine+PDTC group, and the difference was statistically significant (P<0.05). Conclusion NF-κB signaling pathway may play a cer-tain role in the IL-12 expression in DCs induced by nicotine, which may be one of the mechanisms of airway immune balance of asthma exacerbated by smoking exposure.

Objective To establish a method for isolation of pulmonary artery smooth muscle cells fromn mice by magnetic separation.Methods Adult C57BL/6J mice were injected with agarose containing iron powder from the right ventricle under aseptic condition.The lung tissue was harvested and minced for collagenase digestion.Then,the pulmonary arteries containing iron powder were separated by magnetic attraction.The pulmonary artery smooth muscle cells were primary cultured,and passaged when the cell fusion was more than 80％.The cultured pulmonary artery smooth muscle cells were determined under microscope and identified by immunofluorescence staining.Results At 1 d after the primary culture,floating microvascular debris was observed under microscope.At 3 d after the culture,long spindle cells were seen to move out of the iron-containing small vessels.At 7-10 d after the culture,the cell morphology showed a spindle and radial shape,and the vascular smooth muscle ceils showed a "peak-valley"-like growth.Immunofluorescence staining of the cultured pulmonary artery smooth muscle cells showed positive smooth muscle myosin heavy chain and positive smooth muscle cell specific protein,smoothelin.Conclusion The mice pulmonary artery smooth muscle cells can be successfully acquired by magnetic separation,especially the distal pulmonary artery smooth muscle cells.

Objective The initiation step in diseases involving pulmonary vascular remodeling,such aspulmonary hypertension (PH),was the proliferation of pulmonary arterial smoothmuscle cell (PASMC).The most common cause of PASMC proliferation ishypoxia.Krüppel-like factor 4(KLF4)was an extensively expressed zinc fingertranscription factor whichparticipatedin celldifferentiation,cell proliferation,and apoptosis by regulating the expression ofdownstream target genes.In this study,we aimed to clarify whether KLF4inhibits hypoxia-induced PASMC proliferation and phenotypic modulation in the process of pulmonary vascular remodeling.Methods Firstly,rat PASMCs were isolated from rat pulmonary artery and incubated with low-serum SMBM(2% FBS,1% SMCGS) under normoxia (21%O 2 ,5%CO 2 ,74% N2)and hypoxia(5% O 2 ,5%CO 2 ,90% N2)for 24 h,48 h and 72 h,respectively.Cell proliferation was quantified by cell counting kit-8 (cck-8) assayand flow cytometry.We also examined PCNA,SM α-actin,klf4 expression by real-time polymerase chain reaction (PCR) and Western blot. Secondly,the alternation of klf4 acetylation levels was detected byco-immunoprecipitation after hypoxicexposure.PASMCs were transfected with KLF4 expression construct by liposome-mediated gene transfer method,and then assessed the effect of KLF overexpression on PASMC proliferation and differentiation marker gene expression.Results The results of cck-8 and the PCNA expression showed that proliferation of PASMCs cultured in 5% oxygen was inhibited after 24 h but was increased by 72 h inculture.Cell cycle analysis showed that the S cell population was increased afterhypoxic exposure for 72 h.Real-time PCR and Western blot showed that cultured in 5% oxygen resulted indecreased level of SM α-actin for 24 h,and increased level for 72 h in PASMCs.The results of real-time PCR,co-immunoprecipitation and Western blot assaysshowed that the expression levels of KLF4 were up-regulated drastically in PASMCs after hypoxic exposure for 24 h,after 48 h the expression level of KLF4 went down,this process involved the acetylation of KLF4.Cell cycle analysis showed that the number of PASMCs distributed in S phase was decreased and in G1 phase was increased compared with untransfected group after hypoxic exposure for 72 h.The results of real-time PCR and Western blot assays showed that overexpression of KLF4 decreased the expression of SM α-actin in PASMCs cultured in 5% oxygen for 72 h.Conclusions The dedifferentiation and proliferation of PASMC aren′t synchronized underhypoxia, this process involves the acetylation of KLF4.Overexpression of KLF4 inhibites hypoxia-induced proliferation of PASM in vitro.

慢性阻塞性肺疾病(chronic obstructive pulmonary disease,慢阻肺)是一种复杂的全身性疾病,不同患者间的病因及发病机制、临床表现、肺功能、影像学表现、生物标志物乃至预后均存在着显著差异,慢阻肺的各种表型由此产生.“发掘慢阻肺临床终点预测性替代指标纵向研究”(evaluation of COPD longitudinally to identify predictive surrogate end-points,ECLIPSE)是一项始于2008年,涉及欧美多个国家的大规模多中心前瞻性队列研究,收纳样本达近3000例,旨在通过3年的观察随访,发现可提示慢阻肺病情进展的临床及影像学特征、生物标志物、基因表达,从而确立慢阻肺表型并指导个体化治疗[1].目前已发表60余篇期刊文献和70余篇国际会议文献,在慢阻肺表型研究领域具有里程碑性的意义.本文就ECLIPSE成果及意义进行综述.

Objective In this study,with hypoxia-induced pulmonary hypertension rats model,to investigate the expression variation of ERK1/2,p38MAPK in rat pulmonary arteries.Methods Establish the rat model of pulmonary hypertension with sixty male SD rats which were randomly divided into a normal control group,hypoxic exposure 1 day,3 days,7 days,1 4 days and 2 1 days groups and with 1 0 rats in each group.The expression of ERK1/2,p38MAPK protein in rat pulmonary arteries of each group was detected by immunohistochemistry.Results ①RVSP and RV/(LV+S) increased significantly than normal control groups (P<0.05),rats pulmonary vascular thicking after chronic hypoxia 3 d,7 d,14 d,and 21 d.②ERK1/2, p38MAPK protein were widely distributed in pulmonary arterial endothelial cells,smooth muscle cells and fibroblasts.And the longer of the time of hypoxic exposure,the more of protein expression.Conclusions ERK1/2,p38MAPK protein expression is involved in the development of pulmonary vascular remodeling of chronic hypoxic-induced rat PH.

Objective To explore the influence of cigarette smoking on the expression and the activity of calcineurin in asthmatic rat lungs by investigating the expression of mRNA and protein and the activity of calcineurin in rat lungs of cigarette smoking combined asthma group .Methods Thirty‐six Wistar rats were randomly divided into three groups:control group ,asthma group and cigarette smoking combined asthma group .The rats were sensitized and challenged with ovalbumin (OVA) or both tobacco smoke and OVA to establish the model .The following parameters were measured:airway inflammation by HE staining ,the bronchial wall thickness (WAt/Pi) by computer‐assisted image analysis system .The expression of mRNA and protein and the activity of CaN were detected by quantitative real time PCR , Immunohistochemistry and biochemical method respectively .Results ① The bronchial wall thickness of the cigarette smoking combined asthma group was significantly increased compared with that of the control group and the asthma group and it was higher in the asthma group than that in the control group (all P < 0 .01) .② The expression of CaN mRNA and protein and the activity of CaN in the cigarette smoking combined asthma group were higher than those in the control group and the asthma group ,and they were higher in the asthma group than those in the control group(all P < 0 .05) .③ There was a positive correlation between the activity of calcineurin in rat lungs and the bronchial wall thickness ( r =0 .652 ,P <0 .01) .Conclusions Cigarette smoking might play an important part in the development of the chronic airway inflammation and the airway remodeling in asthma by promoting over‐expression of CaN mRNA and protein and increasing the the activity of calcineurin in asthmatic rat lungs .

Pulmonary fibroblasts have key roles in the formation and maintenance of lung structure and function, and are involved in tissue repair and remodeling. Transforming growth factor-beta 1 (TGF-beta 1) induces differentiation of fibroblasts into myofibroblasts, the key effector cells in fibrotic states, which are characterized by the expression of a-smooth muscle actin (alpha-SMA) markers. 1 alpha,25-Dihydroxyvitamin D-3 [1,25(OH)(2)D-3] has been implicated in regulating differentiation, and the vitamin D receptor (VDR) may be a regulator of TGF-beta signaling. In addition, there is presently only limited information regarding microRNA (miRNA) regulation of lung fibroblast differentiation. To determine the role of 1,25(OH)(2)D-3 in regulating the differentiation of fibroblasts induced by TGF-beta 1 and the functional importance of miR-27b, cell culture systems, cell transfection and the 3' untranslated region (3'UTR) luciferase assay were employed. 1,25(OH)(2)D-3 inhibited differentiation and downregulated miR-27b expression in human lung fibroblasts induced by TGF-beta 1. In addition, human lung fibroblasts were transfected with miR-27b mimic or miR-27b inhibitor, and demonstrated that the overexpression of miR-27b decreased the VDR protein expression and increased the expression of a-SMA, while reducing levels of miR-27b had opposing effects. Finally, the luciferase reporter assays were performed to confirm that miR-27b directly targeted VDR 3'UTR. Taken together, these results suggest that 1,25(OH)(2)D-3 inhibits lung fibroblast differentiation induced by TGF-beta 1 via miR-27b targeting VDR 3'UTR, which may be used as a novel treatment strategy in differentiation pathways.

Objective To retrospectively analyze the clinical features ,diagnosis and therapy of pulmonary embolism for reduction of misdiagnosis and missed diagnosis, and for improving the cure rate.Methods The data of related risk factors, clinical manifestation, auxiliary examination results and treatment of 208 patients with pulmonary embolism in Shanxi Province people’s hospital were summarized.Results①Pulmonary embolism in patients with clinical manifestations of diversity, malnly include dyspnea, cough, chest suppress, chest paln and swelling of lower limbs.②The maln risk factors related to pulmonary embolism are deep vein thrombosis, high blood pressure, smoking and cardiac insufifciency.③Routine inspection project D-dimer determination can be used as screening sensitive but not speciifc indicator of pulmonary embolism.④Thrombolysis sequential anticoagulation compared with pure anticoagulant therapy cure rate was statistically signiifcant difference in, there was no statistically signiifcant difference in total effective rate.Conclusions clinical manifestation of pulmonary embolism and routine inspection lack of speciifcity, and should be considered in patients with suspected diagnosis, increase the rate of diagnosis, treat actively.

Objective To observe the expression of basic fibroblast growth factor (bFGF) and matrix metalloproteinase-12 (MMP-12) in pulmonary vascular wall induced by cigarette smoke exposure in rats,and to explore their role in pulmonary vascular remodeling.Methods 40 male SD rats were randomly divided into control group,smoke exposure for one month group,smoke exposure for two months group,smoke exposure for three months group,with ten rats in each group.The ratio of pulmonary vascular wall thickness and vascular external diameter (WT％),and the ratio of pulmonary vascular wall area and total pulmonary vascular area (WA％) were measured by the image analysis system.The collagen fiber formation was observed by Van Gieson staining.The expressions of bFGF and MMP-12 in pulmonary vascular wall were detected by immunohistochemical staining,the protein expression was reflected by average grey value.Results ①As the extension of smoke exposure time,WT％ and WA％ were increased gradually,there was statistical significance among groups (P ＜0.05).②As the extension of smoke exposure time,bFGF and MMP-12 protein expressions increased,there was statistical significance among groups (P ＜0.05).③ bFGF was positivily correlated with WT％ (r =-0.748,P ＜0.05) and WA％ (r =-0.834,P ＜0.05),MMP-12 was positivily correlated with WT％ (r =-0.565,P ＜0.05) and WA％ (r =-0.504,P ＜0.05),bFGF was positively related with MMP-12 (r =0.738,P ＜0.05).Conclusions By upregulating the expressions of bFGF and MMP-12 protein in pulmonary vascular,cigarette smoke exposure induces the remodeling of pulmonary vascular.

Objective To investigate the effects of cigarette smoke exposure on pulmonary artery pressure and expression variation of peroxisome proliferator-activated receptor γ (PPAR-γ) in pulmonary vasculature and bronchus in rats.Methods Thirty male SD rats were randomly divided into a control group,a smoke exposure for 6 weeks group,a smoke exposure for 12 weeks group, with 10 rats in each group.Mean right ventricular systolic pressure (mRVSP) of each group were measured.The expression of PPAR-γ protein in pulmonary vascular and bronchus of each group were detected by immunohistochemistry.Results ① mRVSP increased significantly in the smoke exposure for 6 weeks group (31.33 ± 14.61) and smoke exposure for 12 weeks group (51.00± 14.81) compared to the RVSP of control group (13.43 ± 3.2) (F =15.47, P ＜0.05, respectively).The mRVSP in the smoke exposure for 12 weeks group increased significantly compared to the mRVSP of smoke exposure for 6 weeks group (P ＜ 0.05).②The expression of PPAR-γ protein in bronchus epithelial cell and pulmonary vascular endothelial cell decreased significantly in the smoke exposure for 6 weeks group[(0.41± 0.01), (0.48 ± 0.08)] and the smoke exposure for 12 weeks group [(0.35±0.01),(0.37±0.07)] compared to control group [(0.52± 0.12), (0.51 ± 0.07)] (P ＜0.05, respectively).The expression of PPAR-γ protein in bronchus epithelial cell and pulmonary vascular endothelial cell both decreased in the smoke exposure for 12 weeks group compared to the smoke exposure for 6 weeks group (F =73.47,58.04, P ＜0.05, respectively).③The expression of PPAR-γ protein in bronchus and pulmonary vascular were respectively negatively correlated with mRVSP (r =-0.725 and-0.664, P ＜ 0.05, respectively).Conclusions By downregulating the expression of PPAR-γ protein in bronchus and pulmonary vascular, cigarette smoke exposure induces the increasing of pulmonary artery pressure in rats.