M-CSF targeting into LCL nucleus behaves as a malignancy promotor

Objective: To investigate the functions of nM-CSF in malignant cells. Methods: recombinant M-CSF was targeted into cell nucleus by employing a eukaryotic expression plasmid vector pCMV/myc/nuc. The constructed plasmid was transfected into cells of EBV transformed lymphoblastoid cell line (LCL). RT-PCR, Western blot and immunofluorescent staining showed that recombinant M-CSF was localized into LCL cell nucleus. The transgenic cells showed elevated proliferation potential, enhanced resistance to apoptosis and increased ability of in vitro migration. Conclusion: Nucleus presenting M-CSF might act as a promoting factor in the processes of cellmalignancy.     

Induction of T-cell immunity against leukemia by dendritic cells pulsed with total RNA isolated from leukemia cells

Objectives To assess the feasibility and efficacy of eliciting leukemia-specific T-cell responses in syngeneic mice in vitro and in vivo using dendritic cells (DCs) pulsed with total RNA from leukemia cells.Methods DCs generated from bone marrow culture in vitro in the presence of combined cytokines were pulsed with cellular total RNA isolated from cultured L615 cells by cationic lipid 1,2-dioleoyloxy-3-(trimethylammonium) propane (DOTAP). T-cell responses were evaluated by in vitro proliferation, and cytotoxicity assay. And in vivo immune protection and proghosis of mice with leukemia were studied.Results DCs pulsed with total RNA isolated from cultured L615 cells (DCs/RNA) were remarkably effective in stimulating L615-specific T-cell response in vitro, but did not cross-react with other leukemia cells from syngeneic mice. Vaccination of naive mice with viable DCs/RNA vaccine was able to partly protect from challenge with a lethal dose of live L615 cells, leading to low leukemia incidence and overall survival prolongation. Statistically significant survival was also observed in a low lethal dose of L615-bearing mice that received treatment using viable DCs∕RNA vaccine alone, suggesting that systemic administration of IL-2 could enhance the anti-tumor efficacy of leukemia RNA/DCs vaccine.Conclusions These data support the use of DCs/RNA vaccine as a feasible and effective route to elicit leukemia immunity against unidentified leukemia-associated antigens for treatment of leukemia-bearing animals.     

联合化疗和放射治疗颅内生殖细胞瘤的远期疗效观察

目的：探讨颅内生殖细胞瘤经过化疗并辅以中低剂量放疗后的远期疗效。方法;对1993－1996年收治的45例颅内生殖细胞瘤患者进行静脉化疗并辅以中低剂量放疗,获得随访39例。其中肿瘤位于松果体区24例,松果体区并鞍区8例,鞍区4例,松果体区并脑室内3例。化疗4d为一疗程,使用药物为长春新碱、甲氨蝶呤、平阳霉素及顺铂。用药后每周化验2次血常规,1次血生化,以观察周围血象变化及心、肝、肾功能,并进行针对性治疗。4周后如血常规及血生化的各项指标均正常,再行第二疗程化疗。两个疗程结束后1个月,在肿瘤病灶局部补充中低剂量（25－35Gy)的放疗。每个疗程间进行CT或MRI检查,观察肿瘤消退的情况。结果：39例患者随访5－8年,1例为手术加全剂量放疗后2年肿瘤复发,此次化疗进行2个疗程,但化疗后未再补充放疗,4年后肿瘤复发,家属放弃治疗,患者死亡。1例化疗、放疗后5年肿瘤复发,再行2个疗程化疗,肿瘤消退80％,目前每半年接受1次化疗,病情平稳,生活自理。其余37例（95％）化疗、放疗后5－8年,复查MRI显示肿瘤无复发,患者生存质量良好,其中30例学龄儿童生长发育及智力均未受影响。结论：静脉化疗辅以中低剂量放疗应以治疗颅内生殖细胞瘤的最佳方案。     

中老年人葡萄糖调节受损的临床特征及其与代谢综合征的关系

目的探讨中老年人葡萄糖调节受损(IGR)临床特征及其与代谢综合征(MS)的关系。方法采用横断面调查的方法对重庆局部地区40岁以上2810例自然人群的IGR进行调查,行75g葡萄糖耐量试验后以WHO 1999年IGR诊断标准分组：葡萄糖耐量正常（NGT),IGR[包括空腹血糖受损(IFG),糖耐量低减(IGT),复合性糖耐量低减(IFG IGT)]和糖尿病(DM)。结果IGR检出率18．11％,其中IGT、占IGR的85．26％,IFG占6．68％,IFG IGT占8．06％,与NGT组比较,IGR人群年龄、体重指数(BMI)、血压、甘油三脂(TG)、总胆固醇(TC)、低密度脂蛋白胆固醇(LDL—c)、HOMA—IR等显著增高,高密度脂蛋白胆固醇(HDL—c)、HOMA-B明显较低。与糖尿病组比较,IGR组血压、TG、HOMA—IR升高程度不如糖尿病组显著,HDL—c、HOMA-B降低程度较糖尿病轻。复合性糖耐量低减组和IFG组较IGT组BMI、HOMA—IR明显较高,而HOMA—B明显较低。IGR中高血压、脂代谢紊乱、肥胖或超重及微量白蛋白尿检出率均显著高于NGT组。复合性糖耐量低减组MS检出率高于IGT组。结论重庆局部地区40岁以上人群IGR发生率较高,IGR者伴代谢综合征的发生率高。IGR各亚组存在不同的代谢异常,复合性糖耐量低减组和IFG组较IGT组的BMI、高血压、微量蛋白尿、HOMA—IR增高的程度更高,HOMA—B明显较低。     

抗击严重急性呼吸综合征的医生与护士心理状态的比较与分析

为分析抗击严重急性呼吸综合征(SARS)的医生与护士的心理状态和差异,在SARS暴发的3个月中共收集了来自多家SARS定点医院的1532份医生和护士的心理健康问卷,应用时勘心理量表与Zung自评抑郁量表进行分析比较。发现无论处于一线还是二线,医生的一般心理健康得分要高于护士,自评抑郁量表得分低于护士,且护士的自评抑郁量表得分要高于常模组。逐步相关回归分析显示,抑郁量表得分同压力源,如“社会对医护人员的支持不够”,“治疗条件和生活环境不好”,“缺乏家庭亲人的支持和交流”等有关,同参加体育锻炼的程度,文化程度呈负相关。医生的心理状态和抑郁状态好于护士。提示应针对不同原因给予干预,同时应对于发现的相关因素予以改善。     

重组突变型DNA聚合酶β表达载体pcDNA3.1—polβ的构建

目的：构建含突变型polβ基因重组表达载体。方法：从食管癌标本中提取总RNA,反转录合成cDNA,克隆入pCDNA3．1质粒。以通用鉴定引物PCR扩增、小量提取质粒酶切鉴定重组质粒,并进行测序。结果：测序结果证实重组载体中的外源片段序列与所选取的标本完全相符。结论：成功构建了含突变型polβ基因重组表达载体,可应用于下游研究。     

抑制性消减杂交法构建甲状腺癌差异表达基因文库

目的：筛选并克隆在甲状腺癌组织与正常甲状腺组织之间差异表达的基因。方法：(1)应用抑制性消减杂交(SSH)进行基因差示表达分析,构建人甲状腺癌组织与正常甲状腺组织之间差异表达的cDNA消减文库;(2)克隆、鉴定甲状腺癌组织特异性高／低表达的基因;(3)登录Genbank,运用Blastn程序进行同源性分析。结果：成功构建了高消减效率的人甲状腺癌cDNA消减文库,对其中数个克隆的插入cDNA片段进行测序,经检索Genbank表明,正向SSH产物中有3个片段与来源于结肠上皮腺癌和Lung Epidermoid carcinoma的EST有100％同源性,提示它们来自恶性肿瘤相关基因。反向SSH产物中有5个片段为未知新序列,提示它们可能来自于在甲状腺癌中特异性低表达的新基因。结论：通过抑制性消减杂交技术,构建了人甲状腺癌cDNA消减文库,为下一步筛选鉴定甲状腺癌特异性基因及其全长克隆、功能研究等工作打下了基础。     

红细胞分化去核因子在诱导小鼠红白血病细胞分化以及恶性逆转中的作用

目的：研究小鼠红细胞分化去核分化因子（MEDDF）在红细胞终末分化中的功能。方法：构建在真核细胞中表达的重组质粒pcDNA-MEDDF,用其转染小鼠红白血病细胞系（MEL）,通过分析细胞的生长曲线,分裂指数及在半固体培养基中的集落形成率比较细胞的生长线性,采用RT－PCR检测c-myc及β-珠蛋白（β-globin)基因的表达。结果：转染pcDNA-MEDDF与转染空载体（pcDNA3.1)的细胞相比较,细胞的增殖率减慢,分裂指数降低,在半固体培养其中形成集落的数量减少,转染细胞的联苯胺阳性率达32．8％,用RT－PCR检测发现,转染细胞β-珠蛋白的表达量上升了3．43倍,o-myc表达量下降了66．3％,。结论：MEDDF能命名小鼠红白血病细胞分化并抑制其恶性变。     

PCR扩增抗人视网膜母细胞瘤单克隆抗体轻链可变区基因

目的：获得抗人视网膜母细胞瘤单克隆抗体轻链可变区基因。方法：从分泌抗人视网膜母细胞瘤单克隆抗体的杂交瘤细胞中提取总RNA,逆转录形成cDNA,用扩增小鼠K轻链可变区基因的引物,通过RT－PCR方法,扩增基因,1．5％琼脂糖凝胶电泳鉴定。结果：用轻链引物扩增获得340bp的基因片段。结论;从三株杂交瘤细胞中获得高纯度的总RNA;用RT－PCR方法从一株杂交瘤细胞成功克隆了抗人视网膜母细胞瘤单克隆抗体轻链可变区基因,为基因工程抗体研究奠定了良好基础。     

Brain atrophy in relapsing-remitting multiple sclerosis: fractional volumetric analysis of gray matter and white matter

PURPOSE: To determine the fractional brain tissue volume changes in the gray matter and white matter of patients with relapsing-remitting multiple sclerosis (MS) and to correlate these measurements with clinical disability and total lesion load. MATERIALS AND METHODS: Thirty patients with relapsing-remitting MS and 25 healthy control subjects underwent magnetic resonance imaging. Fractional brain tissue volumes (tissue volume relative to total intracranial volume) were obtained from the total segmented gray matter and white matter in each group and were analyzed. RESULTS: The fractional volume of white matter versus that of gray matter was significantly lower (-6.4%) in patients with MS (P <.0001) than in control subjects. Neither gray matter nor white matter fractional volume measurements correlated with clinical disability in the patients with MS. CONCLUSION: Loss of brain parenchymal volume in patients with relapsing-remitting MS is predominantly confined to white matter. Analysis of fractional brain tissue volumes provides additional information useful in characterizing MS and may have potential in evaluating treatment strategies.