小檗碱对大肠杆菌基因转录抑制作用的实验研究

目的：对小劈碱（BBR）对大肠杆菌的抑制作用的分子靶点进行探讨。方法：鉴于BBR对DNA结合的偏好性,本文从基因转录表达水平对BBR的大肠杆菌生长抑制作用靶点进行实验研究。结果：BBR对于大肠杆菌基因上游调控元件UP element具有较强的亲和力,而在此元件地转录起始区含有TATA碱基序列。进一步对启动子上游含有UP element调控元件的基因sulA、recA、16S和启动子上游不含UP element调控元件的基因lpxC、secG、mutT的mRNA表达比较表明,BBR抑制sulA、recA、16S的表达,对lpxC、secG、mutT无明显作用,提示TATA序列是BBR的作用靶点。结论：本结果对从基因转录水平探讨BBR抑菌作用提供了新思路。     

人参皂苷Rg_1通过作用于炎性因子和糖皮质激素受体减少金葡菌对肺上皮细胞的损伤(英文)

本文旨在探讨人参皂苷Rg_1体外对金葡菌感染肺上皮细胞所致病变的作用及其可能的机制。实验采用体外大鼠原代肺上皮细胞感染模型。RT-PCR和Western blot方法观察肺细胞中integrinβ1,NF-κB和糖皮质激素受体(GR)等相关因子的表达。结果表明人参皂苷Rg_1可以明显抑制金葡菌感染后肺上皮细胞整合素integrinβ1的表达,下调炎性因子NF-κB,ICAM-1,TNF-α,IL-2和IL-6,上调GR的表达。人参皂苷Rg_1可以明显抑制金葡菌感染肺上皮细胞后的炎性因子,从而对肺脏起到一定的保护作用。     

Close correlation between development of MODS during the initial 72 h of hospitalization and hospital mortality in severe fever with thrombocytopenia syndrome

An emerging infectious disease was identified as severe fever with thrombocytopenia syndrome (SFTS) in central China since late March 2009. We found the patients with SFTS had severe clinical symptoms, and progressed rapidly to multiple organ dysfunction syndrome (MODS) with high fatality rate of 25%-30%. The aim of this study was to assess the significance of risk factors predicting the development of MODS and death in SFTS patients. Consecutive SFTS admissions between May 2009 and September 2011 were analyzed for parameters of organ function during hospitalization using Marshall scoring system for MODS, and platelet counts were recorded on admission and at 24, 48, 72 h and one week after admission. We investigated the kinetics of organ failures and analyzed the association between age, platelet count and development of MODS or death. A total of 92 SFTS patients were enrolled in this study. Among them, 32 patients with dysfunction of over 4 organs were identified, 45% of them died within 72 h, 72% died within 5 days, and 76% died within 7 days after admission. We also found cumulative Marshall score was significantly higher in death patients (11.76±2.05) than in survival patients (4.22±1.98) (P<0.001). In addition, SFTS patients had older age and lower platelet counts in MODS and death groups. Furthermore, we also observed that there was a close correlation between platelet count on admission and Marshall score (P<0.001). High Marshall score, advanced age and lower platelet counts were the main risk factors for the development of MODS, and those factors could predict mortality in SFTS patients, suggesting prompt treatment and close monitoring of severe complications, especially MODS, are of great importance in saving patients’ lives.     

IgG-HepG2细胞评价复方苦参注射液及其生产过程可能产生致敏原的研究——利用生物技术进行过程监控致敏性的方法学探索

目的:利用IgG promoter-HepG2细胞对复方苦参注射液生产过程中可能产生致敏原的生产节点进行研究。方法:构建IgG启动子调控绿色荧光蛋白表达质粒,转染入HepG2细胞,与常见致敏性物质(葛根素,卵白蛋白,LPS和伤寒菌疫苗)、复方苦参注射剂中使用到的辅料(NaOH,醋酸,聚氧乙烯脱水山梨醇单油酸酯即吐温80和乙醇)和生产过程中关键节点样品(S1～S8)进行孵育,30 m in后拍照,图像分析软件统计细胞荧光强度变化。结果:IgG promoter-HepG2细胞对常见致敏性物质呈阳性反应。注射剂辅料中醋酸和吐温80有致敏性,生产过程8个样品中,2个样品有显著致敏性。结论:IgG promoter-HepG2细胞对常见致敏原反应敏感,特异性高,适用于中药注射剂中致敏原快速筛选,可实现对中药注射剂生产过程中致敏原的实时监控。     

用于评价致敏原的IgG-HepG2细胞激活过程中相关炎性因子表达的观察

目的 研究转IgG启动子调控GFP基因表达HepG2细胞激活过程中相关炎性因子表达的变化.方法 采用Elisa法评价葛根素和LPS后诱导IgG启动子转基因细胞分泌IL-1β,IL-8,TNF-α和MCP-1蛋白量,qPCR法评价炎性相关因子和固有免疫相关因子的mRNA转录表达量.结果 和HepG2细胞相比,IgG启动子转基因细胞不增加炎性因子分泌和基因表达量,不激活固有免疫.葛根素不增加转基因细胞炎性因子分泌和基因表达量,不激活固有免疫系统.LPS激活固有免疫系统,显著升高IL-8,TNF-α和MCP-1分泌量.结论 IgG启动子转基因HepG2细胞可以作为评价Ⅱ型变态反应的特异性细胞模型,提示葛根素可以作为特异性激活IgG启动子的阳性对照品.     

Expression and Fuactional Role of HERG1, K~+ Channels in Leukemic Cells and Leukemic Stem Cells

In order to investigate the expression and functional role of HERG1 K channels in leukemic cells and leukemic stem cells (LSCs), RT-PCR was used to detect the HERG1 K channels expression in leukemic cells and LSCs. The functional role of HERG1 K channels in leukemic cell proliferation was measured by MTT assay, and cell cycle and apoptosis were analyzed by flow cy- tometry. The results showed that herg mRNA was expressed in CD34 /CD38-, CD123 LSCs but not in circulating CD34 cells. Herg mRNA was also up-regulated in leukemia cell lines K562 and HL60 as well as almost all the primary leukemic cells while not in normal peripheral blood mononuclear cells (PBMNCs) and the expression of herg mRNA was not associated with the clinical and cytoge- netic features of leukemia. In addition, leukemic cell proliferation was dramatically inhibited by HERG K channel special inhibitor E-4031. Moreover, E-4031 suppressed the cell growth by induc- ing a specific block at the G1/S transition phase of the cell cycle but had no effect on apoptosis in leukemic cells. The results suggested that HERG1 K channels could regulate leukemic cells prolif- eration and were necessary for leukemic cells to proceed with the cell cycle. HERG1 K channels may also have oncogenic potential and may be a biomarker for diagnosis of leukemia and a novel potential pharmacological target for leukemia therapy.     

Study on P16 gene in acute leukemia

P1 6gene is located in human chromosome9P2 1 ,the function of which is to suppresscancer.Itencodes 1 6 ku molecular weight nuclear phosphateprotein,regulates activity of cyclin- dependent kinaseK( CDK4) ,controlscellulartransition from G1phaseto Sphae and cellular proliferation.P1 6gene isasso-ciated with the development,progression and malig-nant changes of many,tumors[1] .In this study weexamined changes of P1 6gene expression in acuteleukemia.1　MATERIALSAND METHODS1 .1　 Samples…     

BCL-XL异质性表达与复发急性髓性白血病细胞体外生长类型的关系

选择１５例复发急性髓性白血病（ＡＭＬ）患者,研究ＢＣＬ－ＸＬ的异质性表达与复发ＡＭＬ细胞体外生长类型的关系。用白血病祖细胞培养（ＣＦＵ－Ｌ）判断生长类型,ＲＴ－ＰＣＲ方法检测 ＢＣＬ－ＸＬ ｍＲＮＡ的表达丰度。结果：ＣＦＵ－Ｌ检测中,１０例为生长型,其ＢＣＬ－ＸＬ ｍＲＮＡ ９例高表达,１例为低表达;ＣＦＵ－Ｌ检测 ５例为不生长型,ＢＣＬ－ＸＬ ｍＲＮＡ均为低表达（ Ｐ＜０．０１）。在复发ＡＭＬ中,ＢＣＬ－ＸＬ ｍＲＮＡ的高表达与其细胞体外生长类型有关,提示与其细胞的自分泌生长有关,可以作为复发ＡＭＬ再生耐药的一个判断指标。     

非霍奇金淋巴瘤p53基因突变的研究

迄今为止,ｐ53基因仍是已发现的最重要的抑癌基因,它的失活常导致肿瘤的发生,约50%的实体瘤有ｐ53基因的缺失和(或)点突变,在恶性血液病中ｐ53基因的变化虽不是普遍现象,但和疾病的进展与耐药密切相关,我们已经报道了61例白血病ｐ53基因的变化,现将对恶性淋巴瘤研究的结果报道如下。1　材料与方法1.1　标本来源29例ＮＨＬ石蜡包块为1994年1月至1996年6月门诊及住院患者恶性淋巴瘤活检存档标本,男21例,女8例;经组织病理学确诊并按国际工作组分类标准分型,其中滤泡型ＮＨＬ11例,弥漫型ＮＨＬ18例;免疫学分型为Ｂ细胞性22例,Ｔ细胞性7例。9种人…     

曲古菌素A抑制HL-60细胞端粒酶活性及hTERT的表达并诱导凋亡

目的研究组蛋白去乙酰化酶抑制剂曲古菌素A(trichostatin A, TSA)抑制HL-60细胞端粒酶活性及亚单位hTERT的表达并诱导凋亡的机制。方法采用MTT法和倒置相差显微镜观察不同浓度曲古菌素A对HL-60细胞的抑制作用,流式细胞仪检测600 nmol·L-1TSA作用后的细胞凋亡情况,TRAP-ELISA法检测端粒酶活性变化,RT-PCR分析端粒酶三个亚单位的mRNA表达情况。结果 TSA对HL-60细胞的抑制作用具有时间和剂量依赖性,AnnexinV/PI双染色法检测凋亡显示,600 nmol·L-1TSA作用48 h后,细胞凋亡率为42·6%。600 nmol·L-1TSA作用12、24和48 h后,端粒酶活性分别下降到1·95 ±0·25、1·73 ±0·12和1·52 ±0·09。RT-PCR显示,端粒酶逆转录酶hTERT表达下降,而端粒酶RNA模板(hTR)和端粒酶相关蛋白(hTP1)表达无明显改变。结论 TSA抑制HL-60细胞中端粒酶活性,并诱导凋亡,其机制可能与曲古菌素A下调hTERT转录水平有关。