目的 探究大黄素对肺炎链球菌肺炎大鼠氧化应激损伤及微小RNA-34a/沉默信息调节因子2相关酶1(miR-34a/SIRT1)轴的影响.方法 60只雄性SD大鼠按照随机数字表法分为假手术组、模型组、低剂量(20 mg/kg)、中剂量(40 mg/kg)和高剂量(80 mg/kg)大黄素组,每组12只.采用气管滴加肺炎链球菌法制备肺炎模型,建模成功后24 h,大黄素干预组分别腹腔注射大黄素溶液,每天1次,连续7 d.HE染色观察肺组织病理变化,酶联免疫吸附测定(ELISA)法检测肺组织肿瘤坏死因子-α(TNF-α)、白细胞介素-6(IL-6)和谷胱甘肽过氧化物酶(GSH-Px)水平,可见分光光度法检测超氧化物歧化酶(SOD)、丙二醛(MDA)水平,qPCR检测肺组织miR-34a、SIRT1 mRNA水平,Western blot检测肺组织SIRT1蛋白表达.结果 假手术组大鼠肺组织正常,无明显病理改变;模型组大鼠肺泡塌陷,有大量炎性细胞浸润;低、中、高剂量大黄素组肺组织炎性病变均减轻.与假手术组比较,模型组肺组织TNF-α、IL-6、MDA、miR-34a水平显著增加(P<0.05),肺组织SOD、GSH-Px、SIRT1 mRNA及蛋白水平显著降低(P<0.05).与模型组比较,中、高剂量大黄素组大鼠肺组织TNF-α、IL-6、MDA、miR-34a水平显著降低(P<0.05),肺组织SOD、GSH-Px及SIRT1 mRNA及蛋白水平显著增加(P<0.05),其中高剂量大黄素组变化最明显.结论 大黄素可减轻肺炎链球菌肺炎大鼠炎症反应及氧化应激损伤,可能与调控miR-34a/SIRT1轴有关.

Objective:To analyze the effect of CD100 to monocyte cytotoxicity in non-small cell lung cancer (NSCLC) patients.Methods:Thirty-five NSCLC patients and thirteen healthy controls were included from Zhengzhou Central Hospital between March 2018 and September 2018. Peripheral blood mononuclear cells (PBMC) and bronchial alveolar lavage fluid (BALF) (both tumor site and non-tumor site) was collected from NSCLC patients, while PBMC was collected from healthy controls. Monocytes were purified from PBMC and BALF. Membrane-bound CD100 (mCD100) and CD72 expression on monocytes was measured by flow cytometry. Monocytes from NSCLC patients were stimulated with recombinant human CD100, anti-CD72, matrix metalloproteinase 14(MMP14), or anti-CD100, and were co-cultured with NCI-H1882 cells for 48 h. Tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), granzyme A, granzyme B level in the supernatants, CD16 expression on monocytes, and percentage of target cell death was assessed. Student t test or paired t test was used for comparison. Results:There were no significant differences of peripheral CD14 + mCD100 + percentage, CD14 + CD72 + percentage, CD100 mean fluorescence intensity (MFI), CD72 MFI between NSCLC patients and healthy controls ( P>0.05). CD14 + mCD100 + percentage, CD14 + CD72 + percentage, CD100 MFI, CD72 MFI was remarkably elevated in tumor site compared with in non-tumor site in NSCLC patients ( P<0.05). There was no remarkable difference of peripheral monocytes-induced NCI-H1882 cell death between NSCLC group and control group [(13.95±3.16)% vs (13.22±2.40)%, P=0.451]. Lung-resident monocytes-induced NCI-H1882 cell death was reduced in tumor site when compared with non-tumor site [(11.61±2.81)% vs (14.19±3.57)%, P=0.008 7]. TNF-α, IL-1β, granzyme A, granzyme B level was also decreased in the supernatants of monocytes from tumor site compared with non-tumor site in NSCLC patients( P<0.05). However, there was no statistical difference of CD16 level between two groups( P=0.666). Recombinant human CD100 stimulation promoted NCI-H1882 cell death induced by monocytes from tumor site when compared with unstimulated cells ( P<0.000 1). TNF-α, IL-1β, granzyme A, granzyme B level was also increased ( P<0.05). However, Monocytes, which were pretreated with anti-CD72, induced decreased NCI-H1882 cell death and TNF-α, IL-1β, granzyme A, granzyme B secretion in response to recombinant human CD100 stimulation ( P<0.05). Recombinant human MMP14 stimulation decreased CD14 + mCD100 + percentage and increased soluble CD100 (sCD100) level. NCI-H1882 cell death and TNF-α, IL-1β, granzyme A, granzyme B level was elevated when compared with unstimulated cells ( P<0.05). Anti-CD100 administration decreased sCD100 level. NCI-H1882 cell death and TNF-α, IL-1β, granzyme A, granzyme B level was elevated when compared with MMP14 stimulated cells ( P<0.05). Conclusions:CD100 shedding was insufficient in tumor infiltrating monocytes in NSCLC patients, leading to decreased cytotoxicity. MMP14 might elevate cytotoxicity of tumor infiltrating monocytes via promoting CD100 shedding and sCD100 formation.

目的探究灵芝多糖对盲肠结扎穿孔导致的脓毒症急性肺损伤大鼠肺功能及肺组织炎症的影响。方法 SD大鼠随机分成对照组、模型组及灵芝多糖低、中、高剂量（50、100、200 mg/kg）组和乌司他丁组,除对照组外,其余各组大鼠均采用盲肠结扎穿孔法制备脓毒症模型。给予灵芝多糖和乌司他丁进行干预,采用全自动血气分析仪检测大鼠O 2 分压[p（O 2 ）]和CO 2 分压[p（CO 2 ）];取大鼠右肺中叶计算肺组织湿/干质量;采用ELISA法检测大鼠肺泡灌洗液中白细胞介素-6（interleukin-6,IL-6）、IL-1β、肿瘤坏死因子-α（tumor necrosis factor-α,TNF-α）和核因子-κB（nuclear factor-κB,NF-κB）水平;采用苏木素-伊红（HE）染色法检测大鼠肺组织病理变化;采用Western blotting法检测大鼠肺脏组织中Toll样受体4（toll like receptor 4,TLR4）、p65、磷酸化p65（p-p65）蛋白表达水平。结果与对照组比较,模型组大鼠肺组织结构破坏,肺泡间隔明显增厚,有大量炎性细胞浸润,肺组织炎症评分显著升高（P＜0.05）;肺组织湿质量/干质量显著升高（P＜0.05）;p（CO 2 ）显著升高（P＜0.05）,p（O 2 ）显著降低（P＜0.05）;肺泡灌洗液中IL-6、IL-1β、TNF-α和NF-κB水平均显著升高（P＜0.05）;肺组织TLR4和p-p65/p65蛋白表达水平显著升高（P＜0.05）。与模型组比较,灵芝多糖组大鼠肺组织损伤减轻,肺泡完整性较好,炎症细胞浸润减少,肺组织炎症评分显著降低（P＜0.05）;肺组织湿质量/干质量显著降低（P＜0.05）;p（CO 2 ）显著降低（P＜0.05）,p（O 2 ）显著升高（P＜0.05）;肺泡灌洗液中IL-6、IL-1β、TNF-α和NF-κB水平均显著降低（P＜0.05）;肺组织TLR4和p-p65/p65蛋白表达水平显著降低（P＜0.05）,呈剂量相关性。结论灵芝多糖能够通过抑制TLR4/NF-κB信号通路,从而减轻脓毒症大鼠炎症反应并保护肺功能。

目的 探讨白介素(IL)-13对IL-1受体相关激酶M(IRAK-M)敲除哮喘小鼠巨噬细胞活化及Toll样受体/白介素1受体(TLR/IL-1R)途径的影响.方法 实验分组:对照组、IRAK-M敲除组、哮喘组、IRAK-M敲除+哮喘组、哮喘+抗IL-13抗体组、IRAK-M敲除+哮喘+抗IL-13抗体组.收集肺泡灌洗液(BALF)并对细胞总数、嗜酸粒细胞、中性粒细胞、淋巴细胞、巨噬细胞计数;酶联免疫吸附(ELISA)检测BALF中IL-13、干扰素-γ(IFN-γ)、IL-4表达水平;实时荧光定量PCR(qRT-PCR)检测肺组织中甘露糖受体(MR)、精氨酸酶1(Arg1)、诱导性一氧化氮合酶(iNOS)mRNA水平;蛋白免疫印迹检测肺组织中IRAK-M、IL-13、TLR4、核转录因子kappaB (NF-κB)蛋白水平.结果 与对照组相比,哮喘组各细胞数,BALF中IL-13、IL-4水平,肺组织中MR、Arg1、iNOS mRNA水平及IL-13、IRAK-M、TLR4、胞核NF-κB蛋白水平升高,差异有统计学意义(P＜0.05);BALF中IFN-γ水平,肺组织中胞浆NF-κB蛋白水平降低,差异有统计学意义(P＜0.05).分别与IRAK-M敲除+哮喘组、哮喘+抗IL-13抗体组相比,IRAK-M敲除+哮喘+抗IL-13抗体组各细胞数,BALF中IL-13、IL-4水平,肺组织中MR、Arg1、iNOS mRNA水平及IL-13、IRAK-M、TLR4、胞核NF-κB蛋白水平降低,差异有统计学意义(P＜0.05);BALF中IFN-y水平,肺组织中胞浆NF-κB蛋白水平升高,差异有统计学意义(P＜0.05).结论 抗IL-13抗体与IRAK-M敲除功能类似,均可缓解哮喘小鼠巨噬细胞活化状况.

目的 研究汉防己甲素片联合吡非尼酮胶囊治疗肺间质纤维化的疗效.方法 选取2017年8月—2019年8月来郑州大学附属郑州中心医院治疗的80例肺间质纤维化患者为研究对象,所有患者随机分为对照组和治疗组,每组各40例.对照组患者口服吡非尼酮胶囊,200 mg/次,在两周时间内每次增加200 mg最后达到每次维持剂量为600 mg/次,3次/d.治疗组在对照组基础上口服汉防己甲素片,2片/次,3次/d.两组患者持续治疗2个月.观察两组的临床疗效,比较两组临床症状缓解时间、肺功能指标、血清转化生长因子-β(TGF-β1)、血管内皮生长因子(VEGF)、胰岛素生长因子-I(IGF-I)、Ⅲ型胶原(Ⅲ-C)、Ⅳ型胶原(Ⅳ-C)、透明质酸(HA)水平.结果 治疗后,治疗组总有效率95.00％,显著高于对照组的77.50％(P<0.05).治疗后,治疗组胸憋、喘息、气短、咳嗽、咳痰等症状缓解时间明显短于对照组(P<0.05).治疗后,两组患者最大呼气流量(PEF)、用力肺活量(FVC)、一秒用力呼气容积(FEV1)水平显著升高(P<0.05),且治疗组患者PEF、FVC、FEV1水平升高较多(P<0.05).治疗后,两组TGF-β1、VEGF、IGF-I水平显著降低(P<0.05);并且治疗组TGF-β1、VEGF、IGF-I水平降低较多(P<0.05).治疗后,两组Ⅲ-C、Ⅳ-C、HA水平显著降低(P<0.05);并且治疗组Ⅲ-C、Ⅳ-C、HA水平降低较多(P<0.05).结论 汉防己甲素片联合吡非尼酮胶囊治疗肺间质纤维化具有较好的治疗效果,能够改善患者肺功能,缩短临床症状缓解时间,调节血清炎性因子水平,安全性较高,值得在临床上推广应用.

Objective: To investigate the effect of interleukin (IL)-7 on anti-tumor activity of CD8(+) T cells in non-small cell lung cancer (NSCLC) patients. Methods: Twenty-seven NSCLC patients and ten healthy controls were included from Zhengzhou Central Hospital between January 2017 and July 2017. Plasma, peripheral blood mononuclear cells (PBMCs), and bronchial alveolar lavage fluid (BALF) (both tumor site and non-tumor site) were collect from NSCLC patients, while plasma and PBMCs were also collected from healthy controls. IL-7 level and IL-7 receptor α (CD127) mRNA relative expression was measured. Purified CD8(+) T cells and primary NSCLC cells were stimulated with recombinant IL-7. Cellular proliferation, cytokines secretion, and protein expression in IL-7 signaling pathway were investigated. Direct/indirect contact coculture system of CD8(+) T cells and primary NSCLC cells was also used to assess the cytolytic and noncytolytic activity after IL-7 stimulation. Results: Plasma IL-7 level was significantly reduced in NSCLC patients compared with normal controls[1 731 (1 364, 2 536) vs 2 686 (1 692, 4 786) ng/L, P=0.023). IL-7 level in BALF isolated from tumor site was also remarkably down-regulated compared with non-tumor site in NSCLC patients[1 045 (562, 1 550) vs 1 599 (1 166, 2 107) ng/L, P=0.006 9). There was no statistical difference of CD127 mRNA between the two groups. Recombinant IL-7 stimulation did not affect cellular proliferation and cytokines production in primary NSCLC cells, however, remarkably increased cellular proliferation, interferon (IFN)-γ/tumor necrosis factor (TNF)-α secretion, and signal transducers and activators of transcription 5 (STAT5) phosphorylation and suppressor of cytokine signaling 3 (SOCS3) expression of CD8(+) T cells from NSCLC patients. IL-7 stimulation also significantly enhanced the cytolytic and noncytolytic function of CD8(+) T cells on primary NSCLC cells. Conclusion: IL-7 enhances anti-tumor activity of CD8(+) T cells in NSCLC patients.