基于生物信息学结合实验验证的常春藤皂苷元抗肝癌机制研究

目的 应用生物信息学与实验相结合的方法,分析常春藤皂苷元（hederagenin,HD）抗肝癌的作用机制。方法 通过UCSC Xena数据库获取肝癌RNA-seq数据,应用R软件“Limma”包筛选肝癌差异基因,SwissTargetPrediction数据库预测HD作用靶点,取二者交集基因,String数据库构建交集基因的蛋白质-蛋白质相互作用（protein-protein interaction,PPI）网络,Cytoscape＿3.9.0软件筛选PPI网络中核心基因,采用分子对接方法验证HD与核心基因的结合位点。采用MTT法观察HD对人肝癌HepG2细胞增殖的影响;采用Western blotting法检测HD对HepG2细胞和H22荷瘤小鼠模型瘤体内细胞分裂周期因子25A(cell division cycle 25A,CDC25A)、CDC25B、白细胞介素-6(interleukin-6,IL-6)、雄性激素受体（androgen receptor,AR）、雌激素受体1(estrogen receptor 1,ESR1)和前列腺素内过氧化物酶2(prostaglandin...     

NirB启动子调控下鼠沙门氏菌体内诱导型表达载体的构建

目的构建遗传稳定性良好的沙门氏菌体内诱导型表达载体。方法以克隆载体pGB2为基础,将沙门氏菌厌氧启动子PnirB和EGFP基因串联,并在其多克隆位点MCS下游引入rrnbT1T2转录终止序列,构建沙门氏菌低拷贝体内诱导型表达载体pGnirB-EGFP-rrnb,电转化入鼠伤寒沙门氏菌phoP/phoQ株,对质粒的稳定性及蛋白表达情况进行检测。结果含有低拷贝重组质粒的沙门氏菌在缺失抗生素选择压力下盲传100代后质粒稳定性高于95%,在厌氧静置72 h培养后激光共聚焦显微镜下可观察到明显绿色荧光。结论高度稳定的沙门氏菌体内诱导型表达载体构建成功,为研制以鼠伤寒沙门氏菌为活载体的新型口服疫苗奠定了基础。     

褪黑素对大鼠肝微粒体细胞色素P450酶亚型CYP2C9、CYP2D6、CYP3A4活性的影响

目的 观察褪黑素对大鼠肝微粒体内细胞色素P450 (CYP450) 酶亚型CYP2C9、CYP2D6、CYP3A4活性的影响.方法 以生理盐水为对照,大鼠灌胃0.54 mg·kg-1·d-1的褪黑素,连续7 d,然后测定其肝微体中CYP2C9、CYP2D6、CYP3A4活性.结果 与对照组比较,褪黑素组CYP2C9、CYP2D6、CYP3A4的活性无变化(P>0.05).结论 褪黑素对CYP450酶 CYP2C9、CYP2D6、CYP3A4活性无影响.     

贯叶金丝桃提取物对人结肠癌HCT8细胞增殖和凋亡的影响

目的:研究贯叶金丝桃提取物对人结肠癌HCT8细胞的增殖及凋亡的影响。方法:采用MTT法检测贯叶金丝桃提取物对HCT8细胞的生长抑制作用;流式细胞仪检测贯叶金丝桃提取物对HCT8细胞凋亡率的影响。结果:6.25～200μg/mL的贯叶金丝桃提取物对HCT8细胞生长具有一定的抑制作用,呈现作用时间和剂量的依赖关系;细胞凋亡率随药物浓度升高而逐渐递增。结论:体外实验表明贯叶金丝桃提取物对人结肠癌HCT8细胞具有一定的生长抑制作用,作用机制与诱导HCT8细胞凋亡有关。     

骨金散对去卵巢大鼠Ⅰ型胶原交联N-端肽、Ⅰ型胶原mRNA表达的影响

目的：复制去卵巢大鼠骨质疏松动物模型,探讨中药骨金散对去卵巢雌性大鼠血清Ⅰ型胶原交联N-端肽（NTX）和骨组织Ⅰ型胶原mRNA表达的影响。方法：6月龄雌性SD大鼠40只,随机分为去势组和假手术组。去势组切除双侧卵巢,假手术组切除部分脂肪。大鼠去势后1个月,将去势组随机分为3组：模型组、雌激素组和骨金散组,雌激素组每日灌服己烯雌酚0.05m g/kg,骨金散组按1.8g/kg剂量灌服骨金散,其余各组灌服等量生理盐水。2个月后,酶联免疫吸附法测量血清碱性磷酸酶（ALP）、NTX水平,RT-PCR方法检测右侧股骨骨组织Ⅰ型胶原mRNA表达水平。结果：与假手术组比较,模型组血清ALP、NTX含量明显升高（P〈0.01）,Ⅰ型胶原mRNA水平显著下降（P〈0.01）。雌激素组和骨金散组大鼠体内的ALP、NTX水平显著低于模型组（P〈0.01）,骨金散组大鼠体内的ALP水平高于雌激素组（P〈0.05）。雌激素组和骨金散组实验大鼠骨组织中Ⅰ型胶原蛋白mRNA表达含量明显高于模型组（P〈0.01）。结论：骨金散可减轻去卵巢大鼠ALP、NTX水平的提高,上调骨组织Ⅰ型胶原mRNA的表达水平,这可能与其治疗骨质疏松的机制有关。     

旋毛虫肌幼虫排泄分泌物激活的巨噬细胞和成肌细胞共培养对成肌细胞分化的影响

目的 体外研究与旋毛虫肌幼虫排泄分泌物(ML ES)作用后的巨噬细胞共培养对成肌细胞分化的影响,为深入了解旋毛虫包囊形成机理提供新的思路。方法 通过共培养技术对旋毛虫ML ES激活的巨噬细胞与成肌细胞进行共培养,建立J774A.1-C2C12细胞共培养模型。细胞免疫荧光法以及Western blot分析与旋毛虫ML ES处理的巨噬细胞共培养对成肌细胞分化以及成肌调节因子表达的影响。结果 细胞免疫荧光以及Western blot表明,与对照组比较,成肌细胞与ML ES作用后的巨噬细胞共培养明显降低成肌细胞分化调节因子(MyoD、Myogenin)阳性细胞的数量及终末分化标记物MHC的表达。结论 旋毛虫ML ES可以直接通过调节巨噬细胞活性来影响成肌细胞的分化,为研究旋毛虫与宿主细胞及旋毛虫感染后,宿主细胞间相互作用复杂机制提供了新的思路。     

两种术式治疗老年股骨转子间骨折的疗效比较

目的 探讨股骨近端钛丝固定、人工股骨头置换术与亚洲型股骨近端防旋髓内钉(APFNA)固定术治疗老年股骨转子间骨折的疗效.方法 将62例老年股骨转子间骨折患者按照术式不同分为置换组(采用股骨近端钛丝固定人工股骨头置换术治疗,26例)和APFNA组(采用APFNA固定术治疗,36例).比较两组手术时间、术中出血量、术后下床...     

白血病患者造血干细胞移植前单次全身照射的预后分析

Objective To analyze the prognostic factors of patients with leukemia treated with single fraction total body irradiation (SFTBI) followed by hernatopoietic stem cell transplantation (HSCT).Methods From January 2001 to September 2008, 102 patients received HSCT. The differences of the survival rate, relapse rate and incidence of interstitial pneumonia (IP) between groups regarding different genders, ages, pathological types, transplantation methods and TBI parameters were compared and the factors related with the survival rate, relapse rate and incidence of IP were analyzed. Results The followup time ranged from 15 to 1482 days (median, 406 days). The follow-up rate was 95.1%. 86 and 55patients were followed up more than one year and three years. The 1-and 3-year survival rates were 59.0%and 44.0%. In univariate analysis, the 3-year survival rate was signifcantly different between the groups with and without relapse before transplantation (20% vs. 55%, χ2 = 6.33, P = 0. 012), allogeneictranplantation versus autologous tranplantation (39% vs. 68%, χ2 = 8.06, P = 0.005), grade 3 or more acute graft versus host disease (aGVHD) and grade 0 -2 aGVHD (0% vs. 54%, χ2 = 7.52, P = 0.006),with and without relapse after transplantation (19% vs. 58%, χ2 = 10.13, P =0.001), with and without IP (23% vs. 58%, χ2 =8.35, P=0.004). Multivariate analysis showed that grade 3 or more aGVHD was the only statistically significant prognostic factors (χ2 = 12. 74 ,P =0. 000). The l-and 3-year relapse rateswere 30. 0% and 50. 0%. The incidence of relapse was obviously higher in the group with relapse before transplantation than that without (47% vs. 16%, χ2 =7. 32, P=0. 007). Multivariate analysis showed thatrelapse before transplantation was a significant factor predicting relapse after transplantation (χ2 = 9. 39,P =0. 020). The cumulative incidence of IP was 35.0%. The incidence of IP was different between groups with dose homogeneity > 3% and ≤ 3% (27% vs. 4%, χ2 = 5. 21, P = 0. 023), with and without acute parotitis (34% vs. 3%, χ2 = 14. 15, P= 0.000), allogeneic transplantation group and autologous transplantation group (31% vs. 8%, χ2= 7.70, P= 0.006). Multivariate analysis showed that transplantation methods, acute parotitis and dose homogeneity were statistically significant factors in predictingIP (χ2 = 10. 08 , 10. 08 and 7.69 , P = 0. 002 , 0. 002 and 0. 010 , respectively) . Conclusions Patients who develop grade 3 or higher aGVHD have poor prognosis. Dose homogeneity influences the incidence of IP. Patients undergoing allogeneic transplantation are apt to have IP. Acute parotitis is related with IP and might be a predictor.     

QK—全科广谱治疗仪对冠心病病人血液对氧化反应及红细胞变形？…

本实验测了３５例冠心病病人使用ＱＫ－全科广谱仪治疗前后血中红细胞超氧化物歧化酶（ＯＳＤ）活力;全血谷胱苷肽过氧化物酶（ＧＳＨ－Ｐｘ）,过氧化氢酶（ＣＡＴ）活力;血浆总抗氧化能力（ＡＯＡ）;红细胞膜丙二醛（ＭＤＡ）含量;红细胞滤过指数（ＩＦ）的变化。结果治疗全血ＧＳＨ－Ｐｘ、ＣＡＴ活力、血浆ＡＯＡ明显下降,红细胞膜ＭＤＡ含量明显升高,红细胞ＩＦ明显升高,后与治疗前比较,全血ＧＳＨ－Ｐｘ,ＣＡＴ活力及     

异基因骨髓移植治疗白血病

自1981年至1985年,我们为8例白血病病人进行了异基因骨髓移植。其中7例达持久性植活,且无白血病复发。骨髓移植后存活期最长者已超过56个月,这是中国异基因骨髓移植成功的首批病例。另外一例死于严重的GVHD。8例病人年龄12-37岁,其中急性髓性白血病4例(成人3例系第一次缓解,儿童1例系早期复发)、急淋2例(第一