骨髓间质干细胞异位移植在椎间盘内的迁移及外源基因的表达

目的:观察骨髓间质干细胞异位移植在椎间盘内的存活、迁移情况及外源基因的表达。方法:实验于2001-10/2003-07在西安交通大学环境与疾病相关基因教育部重点实验室完成。选取新西兰大白兔32只,每只兔子的椎间盘均被随机分为4个区组,即正常对照组(L2～3)、生理盐水组(L3～4)、绿色荧光蛋白细胞移植组(L4～5)及BMP-绿色荧光蛋白细胞移植组(L5～6)。分别进行髓核注射,正常对照组不注射;生理盐水组注射20μL生理盐水;绿色荧光蛋白细胞移植组注射20μL林格氏液包含1×105的含绿色荧光蛋白的骨髓间质干细胞;BMP-绿色荧光蛋白细胞移植组注射20μL林格氏液包含1×105的含BMP-绿色荧光蛋白的骨髓间质干细胞。术后1,3,6个月取材,荧光显微镜观察髓核组织内绿色荧光强度;DNA-PCR分析新霉素抗性基因的DNA拷贝数。结果:32只白兔全部进入结果分析,无脱失。①术后不同时间点各组髓核组织内绿色荧光强度比较:绿色荧光蛋白细胞移植组及BMP-绿色荧光蛋白细胞移植组髓核组织内可观察到荧光的存在,随时间的增加,荧光变得比较分散。两组荧光的分布和强度并没有明显的差别。②各组髓核组织内新霉素抗性基因表达水平比较:绿色荧光蛋白细胞移植组及BMP-绿色荧光蛋白细胞移植组术后1,3,6个月组间及组内比较,差异均无显著性意义(P>0.05)。结论:移植的骨髓间质干细胞能够在髓核内存活、迁移,外源基因可以在髓核内表达,提示基于骨髓间质干细胞移植的治疗策略是防治椎间盘退变的潜在有效方法。     

不同发育阶段人表皮干细胞端粒酶反转录酶表达的比较

目的:目前已知端粒酶活性的丧失及其增殖相关基因表达的改变是造成多种成体干细胞体外复制和扩增受限的主要原因,而端粒酶反转录酶对端粒酶活性起关键作用。体外分离培养人胚胎、少儿、成人3种不同皮肤来源的表皮干细胞,对比观察不同发育阶段端粒酶反转录酶表达的差异。方法:实验于2005-09/2007-04在南昌大学第一附属医院烧伤研究所完成。①对象:因创伤等原因致意外流产的妊娠24～26周龄胎儿皮肤标本,4～12周岁少儿及25～45岁成年人烧伤整形植皮手术剩余皮片标本,分别由南昌大学第一附属医院产科、烧伤科提供,产妇与烧伤患者对治疗及实验均知情同意,实验经医院医学伦理委员会批准。②实验方法:取胎儿、少儿和成年人的全层皮肤,用胰蛋白酶和EDTA联合消化法分离表皮,胶原快速贴附法纯化人表皮干细胞,用未黏附的角质细胞作为对照,以含表皮生长因子、角质细胞无血清培养液组成的表皮干细胞培养基进行体外培养。③实验评估:通过β_1整合素、角蛋白19、p63免疫细胞化学染色对培养细胞进行鉴定,计算克隆形成率。以免疫细胞化学染色法和图像定量分析法检测3种不同皮肤来源的表皮干细胞端粒酶反转录酶的表达差异。结果:①细胞生长特征:分离培养的人表皮干细胞呈明显克隆性生长,克隆形成率高于角质细胞对照组(P<0.05)。②表皮干细胞的鉴定:细胞克隆经免疫细胞化学染色后,β_1整合素、角蛋白19、p63均呈阳性表达。③端粒酶反转录酶免疫细胞化学染色及定量表达:不同皮肤来源的表皮干细胞端粒酶反转录酶均呈阳性,但表达强度不同,人胚胎>少儿>成人。3种皮肤来源的表皮干细胞平均吸光度值和阳性面积值均随年龄增加而逐渐降低,人胚胎>少儿>成人(P<0.05)。结论:人胚胎、少儿、成人皮肤来源的表皮干细胞均有端粒酶反转录酶表达,其表达强度依次减弱。提示诱导和增强端粒酶反转录酶的表达对维持表皮干细胞在体外自我更新和增殖能力可能具有重要意义。     

NONMEM法分析临床病人静脉输注阿米卡星的群体药代动力学

收集52例呼吸科感染病人静脉输注阿米卡星(amikacin,AMK)的临床数据,用荧光偏振免疫法(FPIA)测定临床病人AMK的血清浓度275点,按二房室开放模型,用NONMEM程序一步估算出AMK的群体药代动力学参数,并定量地分析病人的体重、肌酐清除率、长期用药和病理状况(慢性阻塞性肺病,COPD)等固定效应对药代动力学参数的影响。经统计分析病人的肌酐清除率和COPD对AMK的药代动力学参数的影响有显著意义,而体重对表观分布容积Vc的影响无显著意义。由最终回归模型估算出AMK的群体药代动力学参数。     

表鬼臼毒素羧酸酯的合成及体外抗肿瘤活性

在三氟化硼·乙醚的催化作用下,鬼臼毒素(I)与有机酸直接脱水生成了具有4β-构型的表鬼臼毒素羧酸酯(IIa～g),所有合成产物经IR,¹HNMR,MS和元素分析等确证其结构,初步药理试验表明,该类化合物对小鼠白血病P388、人胃癌SGC-7901有一定的抑制活性。     

热敏脂质体

目的:综述热敏脂质体的原理和它作为靶向药物载体的应用。方法:论述了脂质体相变的原理和热敏脂质体的研究概况,以及携带化疗药物的热敏脂质体和肿瘤热疗结合后治疗效果的增强作用,指出了热敏脂质体的发展前景。结果:热敏脂质体有良好的热靶向性,在肿瘤治疗方面效果明显。结论:热敏脂质体是新一代热靶向药物载体,有重要的开发价值。     

Squamous cell carcinoma in an ileal neobladder: a case report and review of the literature

Itis rare for a primary tumor to occur in an ileal conduit, although it has been used for urinary diversion for more than four decades. Most malignant tumors that have been reported in ileal conduits have been adenocarcinomas. 1,2 To our knowledge, squamous cell carcinoma in an orthotopic and continent ileal neobladder has not been reported. We report here a case of squamous cell carcinoma and calculi in an ileal neobladder 6 years after a cystectomy and an ileal neobladder diversion for transitional cell carcinoma of the bladder and also review the literature.     

充血性心力衰竭患者甲状腺激素辅助治疗40例

1对象和方法 1.1对象我院2002-05/2004-05住院CHF患者40(男28,女12)例,年龄43～72(63.7±9.6)岁.其中缺血性心肌病25例、扩张型心肌病7例,风湿性心脏病6例,其他2例.心功能为Ⅱ～Ⅳ级(按NYHA分级标准).     

From variome to phenome:Pathogenesis,diagnosis and management of ectopic mineralization disorders

Ectopic mineralization- inappropriate biomineralization in soft tissues- is a frequent finding in physiological aging processes and several common disorders, which can be associated with significant morbidity and mortality. Further, pathologic mineralization is seen in several rare genetic disorders, which often present life-threatening phenotypes. These disorders are classified based on the mechanisms through which the mineralization occurs: metastatic or dystrophic calcification or ectopic ossification. Underlying mechanisms have been extensively studied, which resulted in several hypotheses regarding the etiology of mineralization in the extracellular matrix of soft tissue. These hypotheses include intracellular and extracellular mechanisms, such as the formation of matrix vesicles, aberrant osteogenic and chondrogenic signaling, apoptosis and oxidative stress. Though coherence between the different findings is not always clear, current insights have led to improvement of the diagnosis and management of ectopic mineralization patients, thus translating pathogenetic knowledge(variome) to the phenotype(phenome). In this review, we will focus on the clinical presentation, pathogenesis and management of primary genetic soft tissue mineralization disorders. As examples of dystrophic calcification disorders Pseudoxanthoma elasticum, Generalized arterial calcification of infancy, Keutel syndrome, Idiopathic basal ganglia calcification and Arterial calcification due to CD73(NT5E) deficiency will be discussed. Hyperphosphatemic familial tumoral calcinosis will be reviewed as an example of mineralization disorders caused by metastatic calcification.     

Prognosis of patients with cirrhosis and chronic liver disease admitted to the medical intensive care unit

Patients with hepatic failure admitted to the medical ICU (MICU) generally have a poor prognosis. To determine if there were readily identifiable clinical factors associated with a high predictive value for outcome, we reviewed retrospectively the charts of 100 patients with serious liver disease admitted to the MICU. The overall mortality of the group was 64%. We found that Child's class, a need for mechanical ventilation, and an elevated serum creatinine had the greatest prognostic significance. Ninety-one percent of the patients receiving assisted ventilation, 89% of the patients in Child's class C, and 93% of the patients with creatinine values greater than 1.3 mg/dl died during their MICU admission. Furthermore, a multivariant regression analysis indicated that patients in Child's class C receiving mechanical ventilation who had an abnormal serum creatinine (greater than 1.3 mg/dl) during the first 72 h in the MICU had only a 2% survival rate.