Toxicity to the normal hemocytes by ALA-PDT for the ex vivo purging of hematopoietic stem cell grafts

Objective To study the toxic effects of 5-amionlevulinic acid-based photodynamic therapy (ALA-PDT) on human peripheral blood mononuclear cells (PBMCs), cord blood mononuclear cells (CBMCs) and peripheral blood stem cells (PBSCs), and furthermore, to understand the possible causes of this response. Methods We used MTT assay to detect the survival rate of PBMCs, CBMCs and PBSCs after treated by ALA-PDT under the optimum experiment conditions with U937 as control;Annexin V-FITC/PI was used to detect the pattern of cell death induced by ALA-PDT. By using flow cytometry, we detected intracellular PpIX fluorescence intensity. Results After ALA-PDT treatment the survival rate of PBMCs had no significant change;however in PBSCs and CBMCs, the survival rate reduced to 70%, and the survival rate of leukemia cell U937 was the lowest, about 30%. After incubation with ALA,except for PBMCs, intraceUuiar PplX fluorescence intensity of the other two kinds of normal haemocytes and U937 increased obviously. These results combined with the flow cytometry suggested that the main pattern of cell death here was apoptosts. Conclusion Under the optimum experiment conditions, ALA-PDT has a slight effect on normal haemocytes but excellent depletions of leukemia cells. Therefore, it can effectively purify autologons bone marrow or stem cell grafts.     

DETERMINATION OF POLYDATIN IN POLYGONUM CUSPIDTUM SIEB. ETZUCC. BY TLC-FLUORESCENCE SPECTROPHOTOMETRY

Objective A method of TLC-fluorescence spectrophotometry was established to assay the content of polydatin in polygonum cuspidatum sieb. et zucc. Methods: Polydatin was extracted by methanol and separated with ch‘loroform-acetone-formic acid-water (4:4:0.5:0.2) by thin layer chromatography. The excitation wavelength and emission wavelength were 284 nm and 384 nm, respectively. Results The linear regression equation of the calibrationgraph was y=7.02179x 4.5143, a linear regression correlative coefficient r= 0. 9936. Conclusion This method was proved simple, stable and sensitive. It can be used in quality control of herbs.     

雷诺嗪对兔心房肌细胞快钠电流的影响及使用依赖性阻滞作用

目的 研究雷诺嗪对兔心房肌细胞快钠电流的影响及其是否存在使用依赖性阻滞作用.方法 应用全细胞膜片嵌记录方法,了解雷诺嗪对兔单个心房肌细胞快钠电流的作用,及在不同刺激频率(1Hz、3.3Hz、5Hz)时对快钠电流的影响.结果 30μmmol/L雷诺嗪对兔心房肌细胞快钠电流有明显的抑制作用,其IC_(50)为(25.6±1.8)μmmol/L,并且随着刺激频率的增加其作用加强,存在使用依赖性.结论 雷诺嗪对兔心房肌细胞快钠电流有一定的抑制作用,其存在使用依赖性阻滞作用.     

雄黄治疗维甲酸耐药急性早幼粒细胞白血病-M_3及诱导凋亡的作用

目的　研究雄黄体外诱导人急性早幼粒细胞白血病 M3细胞凋亡的作用 ,并观察其对维甲酸耐药的急性早幼粒细胞白血病 M3的治疗效果。方法　用雄黄处理在体外培养耐维甲酸的人白血病细胞 ,电子显微镜观测白血病细胞凋亡过程 ,并前后对照观察应用雄黄治疗维甲酸耐药的急性早幼粒细胞白血病 M3的疗效。结果　在 1 .0～ 2 .0 μmol·L- 1 的浓度下 ,雄黄作用于白血病细胞 2 4h后 ,可见到较为典型的细胞凋亡形态学变化 :细胞核固缩 ,染色体凝集 ,呈新月型紧贴于核膜周边 ,核碎裂 ,染色质片断化 ,凋亡小体形成等。用雄黄治疗的 2 0例患者 ,完全缓解(CR) 1 7例 ,部分缓解 (PR) 1例 ,总有效率 (CR +PR) 90 %。结论　雄黄能够诱导耐维甲酸的白血病细胞凋亡 ,可用于维甲酸耐药的急性早幼粒细胞白血病 M3治疗 ,且缓解率及长期生存率高。     

THE CHANGE OF SERUM TUMOR NECROSIS FACTOR-α LEVEL AND ITS CLINICAL SIGNIFICANCE DURING THE TREATMENT OF CHRONIC HEPATITIS B WITH LAMIVUDINE

Objective To evaluate the effect of lamivudine on immunity of chronic hepatitis B by observing the sequential changes of serum TNF-α and HBV-DNA level. Methods 31 CHB patients with elevated serum ALT/AST level and HBVDNA level higher than 106 copies/mL were treated with lamivudine (100mg/day) for one year. The sequential serum samples, which were taken at the 0, 3rd, 6th, 12th month after initiation of therapy, were used to detect serum level of TNF-α and quantity of HBV-DNA respectively. Results ① The serum TNF-α levels were higher than normal value before treatment in all patients; ② At In the 3rd month of treatment, The the serum HBV-DNA level began to decline and became negative in the 54.9% of all patients. At the end of treatment, HBV-DNA was negative in 48.4% of all patients; ③ The decrease of TNF-α level was later than HBVDNA level drop. TNF-α level began to decline after 6 months treatment. At the end of treatment, TNF-α level was lower than that at in 6th month, TNF-α level returned to normal in the 38.7% of all patients; ④ The TNF-α level decreased significantly after 6 months treatment in the patients with ALT>80IU/L at the beginning of treatment. But in the patients with ALT≤80IU/L, the TNF-α level decreased just after 12 months treatment; ⑤ TNF-α level fell obviously and early in patients whose HBVDNA became negative at in the 3rd month. Conclusion Lamivudine can suppress the replication of HBVDNA quickly, and decrease TNF-α level in the serum TNF-α level. It suggests that lamivudine can modulate immune response directly or indirectly. The changes of serum TNF-α level may be used to evaluate the clinical efficacy of lamivudine.     

光动力疗法对人乳腺癌细胞MDA-MB-231凋亡的影响

目的探讨光动力疗法(PDT)对人乳腺癌细胞凋亡的影响。方法以人乳腺癌细胞株MDA-MB-231为研究对象,以血卟啉衍生物(HpD)为光敏剂,以630 nm波长激光为光源,采用连续照射的方式。将不同浓度HpD染色的MDA-MB-231细胞,照射不同剂量的激光后,用MTT法测定其A492值,选择最佳作用参数;电镜观察PDT作用后不同时间的细胞凋亡情况,并采用免疫组化方法检测促凋亡蛋白Bax和抗凋亡蛋白Bcl-2的表达情况。结果随着PDT作用后时间的延长,细胞凋亡现象越明显;Bax蛋白表达无明显变化(P>0.05),而Bcl-2蛋白表达自PDT后6 h起显著下降(P<0.01),PDT后6 h Bax/Bcl-2比值依次递增,且有统计学意义(P<0.05)。结论光动力疗法可诱导人乳腺癌细胞发生凋亡,其机制可能与Bax和Bcl-2蛋白表达的比值有关。     

去甲斑蝥素在体外对白血病细胞核酸和蛋白质合成的影响

应用细胞培养、放射性核素示踪技术研究了去甲斑蝥素对白血病细胞的作用。结果表明，去甲斑蝥素对人早幼粒白血病ＨＬ－６０细胞株ＤＮＡ和蛋白质合成有抑制作用，对ＲＮＡ合成未见明显影响；对正常人骨髓细胞ＤＮＡ合成无抑制相反有一定促进作用。     

雄黄对急性早幼粒细胞白血病细胞诱导凋亡和促进分化的双重作用

目的　了解雄黄治疗急性早幼粒细胞白血病 (APL)的机制。方法　以APL细胞株NB4细胞为体外模型 ,应用透射电镜观察细胞形态 ,MTT检测细胞生长增殖状态 ,流式细胞仪检测细胞凋亡 ,NBT还原试验反映细胞分化能力 ,并检测细胞表面分化抗原CD11b、CD3 3 表达的改变。结果　雄黄作用后 ,细胞同时出现凋亡和分化相关的形态改变 ;超微结构发现凋亡小体 ,同时部分细胞内出现分化颗粒 ;流式细胞仪检测线粒体膜表面抗原APO 2 .7表达上调 ;CD11b表达增加 ,CD3 3 表达减少 ,其调变程度弱于全反式维甲酸。结论　雄黄对急性早幼粒细胞白血病细胞同时具有诱导凋亡和促进部分分化的双重效应。     

急性白血病多药耐药蛋白的异常表达及其与临床疗效和预后的关系

目的研究多药耐药蛋白P-糖蛋白(PGP)、多药耐药相关蛋白1(MRP1)、谷胱甘肽-硫-转移酶π(GST-π)和拓扑异构酶Ⅱα(TOPOⅡα)在初发急性白血病(AL)患者中的表达,并探讨其与疗效及预后之间的关系。方法采用免疫组化SABC法检测了PGP、MRP1、GST-π、TOPOⅡα在30例AL患者白血病细胞中的表达情况。结果PGP、MRP1、GST-π、TOPOⅡα在初发AL中表达的阳性率分别为23.33%、36.6%、60%和43.33%;PGP、MRP1阳性组的完全缓解(CR)率均明显低于阴性组(P<0.01);TOPOⅡα阳性组的CR率为92.31%,高于阴性组的58.82%(P<0.05);GST-π阳性组的CR率与阴性组的CR率之间无统计学意义(P>0.05)。结论单一PGP、MRP1的表达升高及TOPOⅡα的表达降低与AL的治疗反应差、预后不良有明显的相关性;联合多个多药耐药蛋白能够更准确地预测AL的耐药和预后。     

人脑胶质瘤nm23-H1蛋白及PCNA的表达

目的　研究人脑胶质瘤中nm2 3 H1蛋白和增殖细胞核抗原 (PCNA)表达状况。方法　采用免疫组化染色技术 ,对 5 3例人脑胶质瘤石蜡切片中nm2 3 H1蛋白和PCNA进行检测。结果　 5 3例人脑胶质瘤免疫组化染色结果显示 ,人脑胶质瘤I级、II级中nm2 3 H1阳性细胞显著高于Ⅲ、Ⅳ级 (P <0 .0 5 ) ;Ⅲ、Ⅳ级中PCNA阳性细胞显著高于I、II级(P <0 .0 5 ) ,但PCNA蛋白表达与nm2 3 H1表达无显著相关性。结论　nm2 3 H1蛋白低表达及PCNA高表达与脑胶质瘤的病理学分级相关 ;nm2 3 H1表达可能成为脑胶质瘤的浸润与转移的指标。     

Tunable Second Harmonic Generation with High Conversion Efficiency in Periodically Poled Lithium Niobate Channel Waveguide

The experiment on quasi-phase-matched second harmonic generation (SHG) in a channel waveguide was reported. The waveguide was made by annealed proton exchange in the periodically poled lithium niobate (PPLN) with the period of PPLN of 14.9 μm, which was designed for cascading wavelength conversion in dense wavelength division multiplexer optical communications. The measurement results of SHG conversion efficiency as a function of fundamental wavelength at room temperature fit well to sinc~2 shape. The peak of SHG conversion efficiency was 75%·W~ -1 ·cm~ -2 as well as reported. The relationship between the center fundamental wavelength and temperature shows that SHG can be effectively tuned by the temperature in PPLN waveguide.     

CD_3 McAb联合rhIL-2激活的骨髓对白血病细胞杀伤净化作用的研究

目的　探讨CD3单克隆抗体 (CD3McAb)联合重组人白细胞介素 2 (rhIL 2 )激活的骨髓对白血病细胞的杀伤及净化作用。方法　用MTT法检测激活骨髓的细胞毒作用 ;半固体集落培养法观察激活骨髓CFU GM水平和对白血病细胞的净化作用 ;采用间接荧光法测定激活骨髓的免疫标记。结果　CD3McAb与rhIL 2联合激活的骨髓对白血病细胞株K5 6 2、HL 6 0均有明显的杀伤净化作用 ,且优于rhIL 2或CD3McAb单用组 (P <0 .0 5或P <0 .0 1)。rhIL 2和 (或 )CD3McAb对激活骨髓的CFU GM水平无明显影响。CD3McAb +rhIL 2激活的骨髓 ,与活化前及rhIL 2或CD3McAb组相比 ,CD3+ 、CD8+ 、CD1 9+ 、CD2 5+ 、CD38+ 、CD56+ 细胞显著升高。结论　CD3McAb能增强rhIL 2激活的骨髓对白血病细胞的杀伤及净化作用     

Study of the mechanism on the apoptosis induced in Human leukemia cell line K562 by the combination of indole-3-acetic acid and horseradish peroxidase

Objective To investigate the mechanisms of apoptosis induced in Human leukemia cell line K562 by the combination of indole-3-acetic acid and horseradish peroxidase. Methods Human leukemia cell line K562 were exposed to indole-3-acetic acid （IAA） at 20, 40, 60, 80 or 100 mol/L and horseradish peroxidase（HRP） at 1.2 g/mL for varying times. MTT assay was applied to detect the cell proliferation. Flow cytometry was performed to detect the arrest of cell cycle. Terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling （TUNEL） assay was used to measure apoptosis. 2, 7-dichlorofluorescin diacetate （DCFH-DA） uptake was measured to determine free radical by confocal microscope. Content of malondiadehyde （MDA） and activity of superoxide dismutase （SOD） were measured by biochemical methods. Results IAA/HRP initiated growth inhibition of K562 cells in a dose- and time-dependent manner. Flow cytometry revealed that cell cycle arrested at G1/G0 after 24 hours treatment. After 72 hours treatment, apoptotic rate of 100 mol/L IAA group increased to 43.9 %, which was 5 times that of control（P〈0.01）. Content of MDA and activity of SOD increased respectively in treatments compared with control. Meanwhile, IAA/HRP stimulated the formation of free radical, which was increased by IAA concentration-dependently. Conclusion The combination of IAAand HRP can inhibit the growth of Human leukemia cell line K562 in vitro by inducing apoptosis which is associated with the increase of free radical. The combination of IAA and HRP might be a promising chemopreventive and chemotherapeutic agent against human leukemia.     

Lasing Frequency Up-Conversion by Using Thermal Population

We theoretically demonstrate a model which can be used to analyze frequency up-conversion of a laser wavelength by using thermal population. The proposed model uses a rate equation model of ytterbium-doped fiber with thermal population effect. The rate and power propagation equations are set up and numerically analyzed to elucidate the dependence of frequency up-conversion efficiency and thermal-optical conversion efficiency on ambient thermal power. The analytical techniques and numerical results show that using pump laser at 1 000 nm,the wavelength can be converted into 900 nm with an up-conversion quantum efficiency of about 99.97% and a cooling efficiency of about 11.1%. This theoretical model is a promising candidate for vast applications in energyefficient laser and energy-utilizing field.     

MCFC GENERATING SYSTEM BY MEANS OF COAL IN CHINA

IntroductionTo generate electricity from the fuel cell is bymeans of utilizing the electrochemistry reaction ofthe combustible gas ( H2 ,CH4etc.) in the fuel cellstack. The coal in solid state can't be directlyused. All the combined cycle generating systemcomposed of the coal as fuel and the fuel cell de-mands to realize the gasification of coal. This pa-per studied the possibility of utilization and devel-opment of the combined cycle generating systemthatis composed of the coal and the molt…     

虎杖提取物白藜芦醇的抗白血病作用及其可能的分子机制

目的探讨白藜芦醇在体外对白血病细胞的抑制作用及其分子机制。方法MTT比色法测定细胞生长抑制率;流式细胞术、HE染色、激光共聚焦显微镜等方法观察各组白血病细胞的细胞周期与细胞凋亡情况;Western Blot-ting法测定细胞抗凋亡蛋白Bcl-2的表达水平;免疫组织化学法测定白血病细胞信号转导分子p-STAT3的活性表达。结果白藜芦醇可以时间-剂量依赖的方式抑制3种白血病细胞增殖、诱导凋亡、影响细胞周期分布、减弱STAT3的酪氨酸磷酸化活性。结论白藜芦醇对不同来源的白血病细胞均具有较强的抗肿瘤活性,显著减少STAT3的酪氨酸磷酸化活性,为其进一步应用于白血病的临床治疗积累了实验依据。     

γ-射线诱导HL-60细胞凋亡的观察

目的 观察γ-射线诱导HL-60细胞凋亡的时间和剂量效应,以探求辐射诱发肿瘤细胞凋亡分子机制。方法 分别以2.5、5、7.5、10、15Gy γ射线照射HL60细胞,于照后继续培养3、6、9、12、24、27、30h,分别收获细胞。电泳检测DNA梯状带;流式细胞术(FCM)细胞凋亡定量并观察细胞周期变化;Hoechst 33258-PI复染荧光显微镜计数凋亡细胞及已死亡细胞百分率。结果 HL-60细胞受照后死亡方式主要是凋亡,其首先阻滞在G２／M期并由此走向凋亡;DNA梯状带及FCM检测结果发现,细胞凋亡具有时间和剂量依赖性;Hoechst 33258PI染色法检测结果与FCM检测的凋亡细胞随剂量的加大及时间的推移而增多的结果并不呈平行关系。结论 γ-射线诱导HL-60细胞凋亡时间和剂量效应关系的确立为进一步探讨该凋亡途径分子机制奠定了基础。     

增强型绿色荧光蛋白标记的Hela细胞株的建立

目的构建增强型绿色荧光蛋白(EGFP)标记的Hela细胞系。方法PCR和DNA测序鉴定pEGFP-C1真核表达质粒,采用Qiagen tip 500进行pEGFP-C1真核表达质粒DNA的提取和纯化;cellfectin转染Hela细胞,G418筛选,有限稀释法单克隆培养,荧光显微镜进行荧光检测EGFP的表达,采用激光共聚焦显微镜观测器检测荧光特性。结果PCR和DNA测序证实pEGFP质粒结构正确;在倒置荧光显微镜下,转染24h后,部分Hela细胞可见绿色荧光,转染72h时,大约80%Hela细胞内均可见绿色荧光。加入含G418的选择性培养基进行选择性培养,选择荧光较强的Hela细胞经有限稀释,持续筛选及克隆化培养,获得稳定表达绿色荧光的Hela细胞克隆,扩大培养并传至10代以上,将此细胞株命名为Hela-EGFP。激光共聚焦显微镜观察发现:在蓝光(-395nm)激发时,Hela-EGFP细胞绿色荧光激发波长为395nm,最大发射峰为509nm。结论Hela-EGFP细胞株具备稳定表达EGFP的能力,为实时可视化进行宫颈癌侵袭转移机制的研究奠定了基础。     

Mirror Mapping of Laser Interferometer Measurement System on Ultra-precise Movement Stage

The accuracy and repeatability of the laser interferometer measurement system （LIMS） are often limited by the mirror surface error that comes from the mirror surface shape and distortion. This paper describes a new method to calibrate mirror map on ultraprecise movement stage （UPMS） with nanopositioning and to make a real-time compensation for the mirror surface error by using mirror map data tables with the software algorithm. Based on the mirror map test model, the factors affecting mirror map are analyzed through geometric method on the UPMS with six digrees of freedom. Dam processing methods including spline interpolation and spline offsets are used to process the raw sampling data to build mirror map tables. The linear interpolation as compensation method to make a real-time correction on the stage mirror unflatness is adopted and the correction formulas are illuminated. In this way, the measurement accuracy of the system is obviously improved from 40 nm to 5 nm.     

紫外线照射对储存血红细胞膜及免疫功能的影响

目的　了解紫外线照射储存血后红细胞在不同时期血液的理化指标及免疫学指标的变化 ,以探讨紫外线照射储存血的安全性及可行性。方法　采用成对血标本对照进行紫外线照射前后及储存 72、12 0h后测定红细胞渗透脆性 ,血浆K+ 、Na+ 、Cl-离子含量和红细胞ATP酶活力及红细胞免疫功能的变化。结果　储存血在保存前后与照射前后红细胞脆性均无显著变化。照射后储存血的红细胞ATP酶活力与对照组相比有所升高 (P <0 .0 5 ) ;红细胞C3 b受体 (RCR)花环率与红细胞C3 b受体花环促进率 (RFER)均有显著升高 (P <0 .0 1)。结论　紫外线照射储存血可以改善红细胞膜的代谢 ,增强红细胞的免疫功能