造血干细胞生物学——研究与展望

造血干细胞（hematopoietic stem cell,HSC）是目前研究方法最为多样、研究技术手段最为成熟的一类组织干细胞,并且已经被成功运用于临床上对白血病以及先天性免疫缺陷等疾病的治疗。近年来,通过对一系列“转基因”与“基因敲除”小鼠模型的分析,人们对造血干细胞在胚胎早期发育过程中的发生与起源、造血干细胞“自我更新”与“定向分化”的调节机制、骨髓中造血干细胞的微环境（niche）对造血干细胞功能维持的调控,以及造血干细胞与白血病干细胞之间的相互关系等诸多方面都取得了很大的进展。如何实现造血干细胞的体外长期培养与扩增,实现胚胎干细胞（embryonic stem cell,ESC）或诱导多能干细胞（induced pluripotent stem cell,iPS细胞）向造血干细胞进行有效的定向分化,以及探索造血干细胞在病理状态（如癌症、贫血、衰老等）或应激状态下（如炎症与感染、组织损伤、代谢异常等）的功能变化,都将会是今后造血干细胞研究的重要方向。     

制霉菌素抗性筛选高活性苎麻脱胶菌诱变菌株

利用制霉菌素抗性筛选高渗透性突变株,提高黑曲霉菌对苎麻纤维的脱胶能力,使微生物脱胶能用于工业生产实践之中.分别用紫外线、硫酸二乙酯、亚硝酸作为诱变剂对黑曲霉3.0.2菌株进行诱变处理.以制霉菌素抗性为遗传标记,从突变菌株中定向筛选得到一株高活性苎麻脱胶菌黑曲霉3.0.2-26.在以未经刮制的苎麻韧皮为主要碳源,0.7%(NH_4)_2SO_4为氮源,添加00.5%KCl;00.5%MgSO_4;0.1%K_2HPO_4; 0.1%酵母膏;Tween80 0.1%的培养液中,接入黑曲霉3.0.2-26,置30℃下,150 r/min处理30 h左右,脱胶苎麻纤维的残胶率平均为14.43%.     

Cloning,expression, and characterization of a novel diketoreductase from Acinetobacter baylyi

Reductions of carbonyl groups catalyzed by oxidoreductases are involved in all biological processes and are often a class of important biocatalyst. In this article, we report a novel enzyme designated as diketoreductase （DKR） that was able to reduce two carbonyl groups in a diketo ester to corresponding dihydroxy ester with excellent stereoselectivity. The DKR was cloned from Acinetobacter baylyi by reverse genetic method, heterogeneously expressed in Escherichia coli, and purified to homogeneity by two chromatographic steps. This novel enzyme exhibited dual cofactor specificity, with a preference of NADH over NADPH. The dihydroxy ester product catalyzed by the DKR was only 3R,5S-stereoisomer with both diastereomeric excess and enantiomeric excess values more than 99.5%. In addition, some biochemical properties of the enzyme, such as the optimal pH and temperature, were also characterized. Furthermore, sequence analysis indicated that this new enzyme was homologous to bacterial 3-hydroxyacyl coenzyme-A dehydrogenase. More importantly, based on the unique catalytic activity and excellent stereoselectivity, the DKR could be utilized in the synthesis of valuable chiral drug intermediates, such as Lipitor.     

非小细胞肺癌三维适形放射治疗靶区和食管剂量学研究

目的:通过剂量学研究明确非小细胞肺癌各照射方法中食管放射损伤.方法:分为常规放疗和三维适形放射治疗(3D-CRT)计划组.3D-CRT分为全纵隔预防照射(TNI)、选择性淋巴引流区预防照射(SNI)、不做选择性淋巴引流区预防照射(N-SNI)三组.通过剂量体积直方图(DVH)、正常组织并发症概率(NTCP)、肿瘤控制率(TCP)评价各治疗计划.结果:常规放疗、TNI、SNI和N.SNI组PTV-2的TCP分别为:0.92、0.95、0.95、0.96;食管修稿≥45Gy照射的百分比(V45)分别为37.45%、48.04%、38.4%、29.67%;NTCP分别为0.0762、0.1142、0.0612、0.0551.结论:3D-CRT在靶区剂量分布上明显优于常规放疗,选择性和不做选择性淋巴引流区预防照射可以更好的减少食管放射性损伤.     

LaminB1、NMP238表达与宫颈癌放疗敏感性的相关性研究

目的:观察LaminB1、NMP238在放疗敏感性不同宫颈癌组织中的表达及意义.方法:收集治疗前宫颈癌组织标本,病理诊断均为中分化鳞癌,在进行放疗后,根据WHO实体瘤放疗疗效判断标准,从中选择40例对放疗敏感的病例(高敏感组)和40例对放疗不敏感的病例(低敏感组),应用分子原位杂交方法检测组织中这些蛋白的表达.结果:免疫组化结果显示LaminB1,NMP238的阳性表达率在高敏感组中均低于低敏感组,LaminB1和NMP238表达具有显著相关性.结论:LaminB1、NMP238表达与宫颈癌放疗敏感性有关.     

CT定位脑立体定向丘脑中央中核毁损治疗癌症顽痛的应用

目的:探讨CT定位脑立体定向丘脑中央中核毁损治疗癌症顽痛的治疗效果.方法:应用立体定向技术,在CT直接靶点定位下,对62位癌痛顽痛进行双侧的中央中核毁损,采用WHO推荐的10分法评定手术前与术后即时及术后3月的疼痛程度.结果:本组患者术前疼痛评分均在7分以上,术后除2例死亡外,其余均降至0～3分,与术前相比有显著差异(P＜0.01).3个月内追踪观察26例患者疼痛有不同程度的复发,但疼痛评分均≤7分,与术前相比有显著差异(P＜0.05).并发症:颅内靶点出血2例,嗜睡4例,偏瘫3例,尿失禁3例,构音障碍2例,动眼神经麻痹、呃逆感觉异常各1例.经对症处理,绝大多数可以恢复.结论:CT定位立体定向中央中核毁损治疗癌症顽痛,创伤小、疗效可靠,是治疗癌性顽痛较为理想的手术方法.     

Reduced cardiac output is associated with decreased mitochondrial efficiency in the non-ischemic ventricular wall of the acute myocardial-infarcted dog

Cardiogenic shock is the leading cause of death among patients hospitalized with acute myocardial infarction (MI).Understanding the mechanisms for acute pump failure is therefore important.The aim of this study is to examine in anacute MI dog model whether mitochondrial bio-energetic function within non-ischemic wall regions are associated withpump failure.Anterior MI was produced in dogs via ligation of left anterior descending (LAD) coronary artery,thatresulted in an infract size of about 30% of the left ventricular wall.Measurements of hemodynamic status,mitochondrialfunction,free radical production and mitochondrial uncoupling protein 3 (UCP3) expression were determined over 24h period.Hemodynamic measurements revealed a>50% reduction in cardiac output at 24 h post infarction when com-pared to baseline.Biopsy samples were obtained from the posterior non-ischemic wall during acute infarction.ADP/Oratios for isolated mitochondria from non-ischemic myocardium at 6 h and 24 h were decreased when compared to theADP/O ratios within the same samples with and without palmitic acid (PA).GTP inhibition of (PA)-stimulated state 4respiration in isolated mitochondria from the non-ischemic wall increased by 7% and 33% at 6 h and 24 h post-infarctionrespectively when compared to sham and pre-infarction samples.This would suggest that the mitochondria are uncoupledand this is supported by an associated increase in UCP3 expression observed on western blots from these same biopsysamples.Blood samples from the coronary sinus measured by electron paramagnetic resonance (EPR) methods showedan increase in reactive oxygen species (ROS) over baseline at 6 h and 24 h post-infarction.In conclusion,mitochondrialbio-energetic ADP/O ratios as a result of acute infarction are abnormal within the non-ischemic wall.Mitochondria ap-pear to be energetically uncoupled and this is associated with declining pump function.Free radical production may beassociated with the induction of uncoupling proteins in the mitochondria.     

MRI定位式全身3D适形放疗的临床应用及体会

目前,放射治疗的主要发展是立体定向放射外科（γ刀,X刀）和三维适形放射治疗（3D CRT）。后者可分为螺旋CT和MRI等多种定位方式。MRI与三维放疗计划系统（3D TPS）联用,其最大优点是通过eflim,可同时获得三维（SAG／TRA／COR）T1 W1、TzWz多层次图像,对于靶区（Tumor）GTV和CTV轮廓勾画的定位精确性明显优于CT,因此具有很高的临床应用价值。现将我们在临床上的一些应用体会总结如下,以供同行们参考。     

重金属 Ni2+ 对亚洲玉米螟生长发育和生殖行为的影响

【目的】探索农田重金属Ni²⁺污染对植食性昆虫潜在的影响。【方法】在人工饲料中添加不同浓度（1, 5, 10, 15和20 mg/kg）的重金属Ni²⁺,以不添加重金属的人工饲料为对照,实验室条件下饲养亚洲玉米螟 Ostrinia furnacalis (Guenée)初孵幼虫,系统观察其各项生长发育和生殖行为指标参数。【结果】Ni ²⁺对亚洲玉米螟幼虫期、蛹期、化蛹率和羽化率有抑制作用;对亚洲玉米螟雌蛹重有促进作用,但随Ni²⁺浓度的升高而逐渐减弱;低浓度Ni²⁺（1~10 mg/kg）对雄蛹重有促进作用,高浓度Ni²⁺（15~20 mg/kg）对雄蛹重有抑制作用。随着Ni²⁺浓度的增加,亚洲玉米螟雌蛾平均求偶持续时间逐渐减小;1 mg/kg Ni²⁺浓度下雌蛾求偶率及求偶高峰期出现的时间与对照组基本一致,其余4个Ni²⁺浓度都对雌蛾求偶行为产生影响且表现为求偶高峰期出现的时间提前,最高求偶率均小于100%。5 mg/kg Ni²⁺对亚洲玉米螟雄蛾定向行为有促进作用,其余浓度则表现为抑制作用。Ni²⁺对亚洲玉米螟雌蛾的产卵量和卵孵化率均产生了显著的抑制作用。【结论】重金属Ni²⁺对亚洲玉米螟的生长发育和生殖行为产生了影响。